Ceroid Pigment Research Articles

Combined use of biomarkers to assess the impact of untreated wastewater from the Danube River, Serbia.

In this study a battery of bioassays, both in vivo (metals and metalloids concentrations, erythrocyte morphometry, comet assay, micronucleus assay, and histopathological analyses) on vimba bream Vimba vimba (L., 1758) and white bream Blicca bjoerkna (L., 1758), and in vitro (treatment of HepG2 cells with native water samples) was applied to assess the harmful potential of untreated wastewater. Faecal indicator bacteria were quantified to assess the microbiological water quality. Vimba bream had significantly higher Fe concentrations in both liver and muscle, while white bream had higher Ca and Cu concentrations in liver. Vimba bream had a significantly higher level of DNA damage in both liver and blood cells, in comparison to white bream. Low levels of micronucleus and nuclear abnormalities were observed in both species. Erythrocytes morphometry did not show significant interspecific differences. Histopathological analyses revealed a similar response of the studied species, with a significantly higher presence of ceroid pigments in the liver of vimba bream. Treatment of HepG2 cells revealed the high genotoxic potential of water downstream of the discharge point. The results of this study clearly demonstrate the importance of effect-based monitoring, in order to enforce more efficient management of natural resources and implementation of wastewater treatment systems.

Schmallenberg virus infection suspected in a calf born to an imported heifer.

Schmallenberg virus infection suspected in a calf born to an imported heifer.

Scanning electron microscopy of Onchocerca fasciata (Filarioidea: Onchocercidae) adults, microfilariae and eggs with notes on histopathological findings in camels

BackgroundOnchocerca fasciata is a prevalent filarial species in camelids of Asia and Africa forming nodules in the skin of dromedary and Bactrian camels. In spite of recent advances in the biology and epidemiology of this nematode species, a relatively scant number of studies have focussed on the morphology of this parasite. The main objective of this study was to describe morphological characteristics of adults, microfilariae and eggs of O. fasciata by scanning electron microscopy (SEM), staining and histology.MethodsFrom April 2016 to March 2017 dromedary camels (n = 456) were inspected for infection with O. fasciata in a slaughterhouse in Kerman (south of Iran). Adult worms in nodules were isolated by digestion of nodules in collagenase and used for SEM. Skin nodules were also fixed, sectioned and stained with hematoxylin and eosin for histopathology. Skin microfilariae that were isolated from tissues surrounding the nodules were confirmed as O. fasciata by sequencing of the cytochrome c oxidase subunit 1 (cox1) and 12S rRNA genes and used for SEM and Giemsa staining.ResultsSingle or multiple O. fasciata nodules (1.2–2.2 cm in diameter and 507–845 mg in weight) were found in 30.3% of the examined camels. SEM analysis helped identify 18 papillae in the caudal region of the male. Discontinuous longitudinal cuticular crests were observed in the posterior region of the male. In female nematodes, the ridges had a rounded shape with a height/width ratio of 7/16 in longitudinal sections. Unsheathed skin microfilariae with a rounded anterior extremity measured 210.7 × 2.5 μm on average. Developed eggs containing microfilariae measured 35.9 × 31.0 μm and their smooth shell surface had characteristic tongue-like appendages. In addition to inflammatory reactions surrounding the parasites, accumulation of intracellular ceroid pigment, golden-yellow to brown in colour, was observed within macrophages upon histopathological examination.ConclusionsWe found longitudinal crests on the surface of the posterior region of the male nematode. Measurements of the main morphological features of microfilariae and eggs, and the shape index of ridges (height/width) in female nematodes are described for the first time.

Osteonecrosis of the femoral head in people living with HIV: anatomopathological description and p24 antigen test.

ObjectiveTo examine the presence of HIV in bone tissue of people living with HIV (PLWHIV) with osteonecrosis of femoral head and describe clinical and anatomopathological findings.DesignThis is a case series which included 44 PLWHIV with osteonecrosis of femoral head who underwent total hip arthroplasty.MethodsClinical data were obtained through analysis of the patients’ medical records. Bone tissue obtained during total hip arthroplasty was retrieved and sent for conventional and immunohistochemical analysis. Monoclonal antibodies were used to mark the p24 (HIV), CD31 (vascular endothelial cells), CD68 (macrophages), and D240 (cells of the lymphatic endothelium) antigens.ResultsDyslipidemia was found in 48% of the patients and lipodystrophy in 31%. Histological analysis showed similar characteristics for the entire sample. Degeneration of joint cartilage was visualized with the presence of fissures and fibrillations, as well as subchondral sclerosis and necrosis of the subchondral cancellous bone tissue. Lymphoplasmocytic inflammatory reaction was observed, with the presence of macrophages containing a foamy, vacuolated cytoplasm, as well as the presence of ceroid pigment and occasional granulation tissue. The reaction with the monoclonal anti-p24 antibody was negative in the samples from all 44 PLWHIV undergoing hip arthroplasty. Reactions with the anti-CD31 and anti-D240 antibodies were negative. Staining with CD68 antibody confirmed that the cells visualized with foamy, vacuolated cytoplasm were macrophages.Conclusionp24 HIV antigen was not detected in the bone tissue of PLWHIV and osteonecrosis. The most frequent anatomopathological findings were extensive necrosis of bone tissue, large vacuoles filled with fat cells, inflammatory lymphoplasmocytic reaction with macrophages containing vacuolated cytoplasm, and the presence of ceroid pigment.

A Case of Over-the-Counter, Drug-Induced Autoimmune Hepatitis: Immunohistochemical and Electron Microscopic Studies

Introduction: In daily clinical practice, drug-induced liver injury(DILI) can always be a cause of liver injury in patients taking medications. Over-the-counter(OTC) drug: Pabron has been purchasing at the pharmacy in Japan, Thailand, Malaysia, and Chinese Taipei as a multi-component cold medicine. We desribed a case of DI-autoimmune hepatitis(AIH) from OTCdrug: Pabron gold®. Case description: A 64-year-old man sought treatment for common cold and was admitted for general fatigue and liver dysfunction. Blood test results included total bilirubin (T-bil), 2.4 mg/dL; direct bilirubin (D-bil), 1.4 mg/dL; AST, 1,244 IU/L; ALT 1,455 IU/L; ALP 1,242 IU/L; and γ-GTP 599, IU/L. Viral hepatitis serology was negative. Other relevant data included IgG level, 1,907 mg/dL; IgM, 95 mg/dL; ANA x80 (cut-off, < x40); ASMA x20 (cut-off, OTC, namely Pabron Gold®, prior to disease onset which was suspected as the cause of liver injury. A DI lymphocyte stimulation test for Pabron Gold® was positive, 2,447 cpm (503%) (< 179%). The liver biopsy revealed spotty necrosis and the accumulation of ceroid pigment in Kupffer cells around central vein, and piecemeal necrosis with multiple plasma cells around portal tracts. Moreover, the plasmocytic infiltration showed positive CD79a staining while negative CD20 staining. Ultrastructually, “cart-wheel” chromatin configuration in the nucleus was observed in some of plasma cells packing with a lot of rough-surfaced endoplasmic reticulum. Moreover, the characteristic granular osmiophilic materials and lipid droplet were observed in Kupffer cell. HLA typing test was HLA-DR4 positive. The score based on simplified criteria for autoimmune hepatitis diagnosis was 6, probable. With a persistent rise in aminotransaminase levels, the patient was started on steroids with predonine prescribed to control the hepatic inflammation. Discussion: This report is the first to describe a case of DI-AIH following the use of an over-the-counter drug. In fact, DI-AIH diagnosis represents a challenge to the clinician because neither histological nor clinical pathognomonic features of AIH exist. We described a case of OTC Pabron gold®-associated liver injury, diagnosed as DI-AIH, calling for caution in the use of OTC.

The fuss over lipo"fuss"cin: not all autofluorescence is the same.

Since the first description of cellular autofluorescence over a century ago, we have now come to appreciate that autofluorescence should not be discarded as a biological artifact but embraced as a biological phenomenon with potentially important cellular relevance. Indeed, cellular and tissue autofluorescence has been attributed to a spectrum of unrelated molecules such as porphyrins, vitamins (vitamin A, riboflavin, thiamine), structural proteins, lipofuscin and ceroid pigments. We have recently shown that freshly isolated epithelial cancer stem cells (CSCs) bear autofluorescent vesicles in the cytoplasm. Our studies definitively prove that riboflavin and not lipofuscin is the source of autofluorescence in CSCs as the inhibition of ATP and not autophagy eliminates CSC autofluorescence, that the ATP-dependent transporter ABCG2, for which riboflavin is a substrate, is overexpressed in autofluorescent CSCs and co-localizes with the membrane of intracellular autofluorescent vesicles, the ABCG2-specific inhibitor Fumitremorgin C reversibly eliminates CSC autofluorescence, riboflavin is a substrate for ABCG2, and only the addition of riboflavin to vitamin-deprived CSC cultures is capable of restoring autofluorescence. Thus, the sum of these data unequivocally supports the conclusion that the source of CSC autofluorescence is the vitamin riboflavin.

Kupffer cell structure in the juvenile Nile crocodile,Crocodylus niloticus

The morphology of Kupffer cells was examined in the liver of the juvenile Nile crocodile using light microscopy and transmission electron microscopy. Pleomorphic Kupffer cells were located in the sinusoids, in the space of Disse, in the hepatic parenchyma and often connected adjacent sinusoids. The cell surfaces were irregular due to the presence of filopodia and lamelliapodia with phagocytosis of white blood cells, red blood cells and thrombocytes being evident. The cells were in close contact with endothelial cells and pit cells in the sinusoidal lumen and with stellate cells in the space of Disse. The cytoplasm contained large phagosomes comprising a combination of ceroid pigment, melanosomes and siderosomes. The nuclei were often indented and eccentrically placed due to the presence of the phagosomes. Conspicuous clusters of membrane-bound tubular organelles with a filamentous or crystalline interior were observed in the cytoplasm. The clusters were sometimes separated into smaller groups around phagosomes. A clear zone existed between the limiting membrane and the interior of these tubular organelles with the electron-dense interior profiles being, respectively, circular, angular or divided. The tubular organelles have not previously been described in Kupffer cells and possibly represent lysosomes with specialized functions. Mitochondria, microtubules, Golgi profiles, granular and smooth endoplasmic reticulum, and a few cytoplasmic lipid droplets were also present. The presence of the tubular organelles and the occurrence of the Kupffer cells in different locations in the liver of the juvenile Nile crocodile are indicative of particularly active and mobile cells.

Pansteatitis associated with high levels of polychlorinated biphenyls in a wild loggerhead sea turtle Caretta caretta

We describe the morphologic and toxicological findings in a case of pansteatitis in a stranded loggerhead sea turtle Caretta caretta. At necropsy, a large amount of adipose tissue in the celomic cavity showing very firm, yellow to orange irregular formations was observed. Histological lesions ranged from the infiltration of necrotic fat by scarce multinucleated giant cells and numerous macrophages containing xylene-insoluble lipopigment inclusions to the presence of several granulomas characterized by an irregular central necrotic area consisting of lipopigment surrounded by numerous multinucleated giant cells. Microbiological cultures were negative. Celomic fat was analyzed for PCBs and DDTs, resulting in very high levels of PCB 138, 153, 180 209 (3170, 2830, 980 and 1190 ng g-1, respectively). Although a nutritional cause cannot be ruled out, the high levels of PCBs detected in the celomic fat could have induced lipid peroxidation in adipocytes, resulting in cell damage, deposition of ceroid pigment and inflammatory response. This is the first report of pansteatitis in a wild sea turtle.

Effects of Aflatoxin on Liver and Protective Effectiveness of Esterified Glucomannan in Merino Rams

The effects of total aflatoxin (AF) given orally on liver in Merino rams were studied. In addition, this study was conducted in order to evaluate the efficacy of an esterified glucomannan (EG) for protection against aflatoxicosis. One-year-old 32 Merino rams were divided into four equal groups. The control group (C) was fed with the commercial feed. The AF group was fed with commercial feed plus 250 μg/day of total AF. The EG group was fed with commercial feed plus 2 g/day of EG. The AF + EG group was fed with commercial feed plus 250 μg/day of total AF and 2 g/day of EG. After feeding period, tissue samples were taken from the liver in order to perform histological analyses. Vacuolar degeneration with small and large droplets and hydropic degeneration in hepatocytes were observed in the AF group. The ceroid pigmentation was observed in macrophages in groups or one by one. It was observed that the fat rate in hepatocytes was 2.6% in the C group, 35.5% in the AF group, 2.9% in the EG group, and 9.6% in the AF + EG group. In conclusion, the adverse effects caused by aflatoxicosis on the liver could be ameliorated by adding EG to the ration.

Preliminary acute and subchronic toxicity studies of GLG-V-13, a novel class III antiarrhythmic agent, in mice.

The acute and subchronic toxic effects of GLG-V-13 (3-[4-(1H-imidazol-1-yl)benzoyl]-7-isopropyl-3,7-diazabicyclo[3.3.1]nona ne dihydroperchlorate, CAS 155029-33-7), a novel class III with some class Ib antiarrhythmic activity, were investigated in mice. The estimated LD50 for GLG-V-13 given orally were 419 mg/kg for male mice and 383 mg/kg for female mice, respectively. The acute toxic signs appeared to be of the central nervous system in origin. Four groups of mice (15 per sex, group and dose) were fed daily with diets containing GLG-V-13 for 90 consecutive days. The equivalent daily doses were 0, 22, 50 and 121 mg/kg/day and 0, 27, 60 and 136 mg/kg/day for male and female mice, respectively. All of the mice survived. Food consumption was decreased. However, mean body weight and body weight gain were not significantly changed. Gross pathological changes, especially in the lungs and liver, were found in the middle and high dose groups. Consistent increased mean corpuscular hemoglobin concentration and decreased mean corpuscular hemoglobin were observed in all dose groups. Hepatocellular necrosis was found in both male and female mice treated with the drug and was dose-dependent. Marked vacuolation of the X zone in the adrenal gland with mild to moderate deposition of ceroid pigments (brown degeneration) was observed in female mice. Lesions in the kidneys and adrenal glands may be a possible reason for changes in serum sodium and potassium ions concentrations leading to an increase in water intake. A significant reduction in cholesterol in the high dose group may be a favorable pharmacological effect of GLG-V-13. The data from the 90-day subchronic toxicity studies indicate that GLG-V-13 appears to have limited systemic toxicity potential.

Feeding fish according to organic aquaculture guidelines EC 710/2009: Influence of potato protein concentrates containing various glycoalkaloid levels on health status and growth performance of rainbow trout ( Oncorhynchus mykiss)

A feeding trial was conducted to evaluate the effect of potato protein concentrate (PPC) replacing fishmeal in nutrition of rainbow trout ( Oncorhynchus mykiss) according to organic aquaculture guidelines. Eight diets were formulated by partial replacement of fishmeal (25%, 50%, 75% and 100%) with PPC of two different qualities (LG-PPC with low glycoalkaloid content of 7.41 mg kg − 1 dry weight, HG-PPC with high glycoalkaloid content of 2150 mg kg − 1 dry weight) and one experimental diet without PPC served as control. Experimental diets were fed over a period of 84 days to triplicate experimental groups until apparent satiation. Best growth performance was observed in the control group. The inclusion of LG-PPC resulted in significantly reduced feed intake, feed utilization and growth with increasing replacement of 25% and higher. Increasing amount of dietary LG-PPC inclusion resulted in alterations in liver histology and in anterior intestine indicating an impact of glycoalkaloids on the different gastrointestinal tissues. The HG replacement revealed most prominently a decreased feed intake and growth parameters indicating massive undernourishment. Congruently, hypotrophy of the liver with ceroid pigment accumulation, and villi degeneration in the intestine were observed in all HG groups. Blood parameters were equally affected by an inclusion of PPC (LG and HG) in the diets involving reduced plasma TG (HG75), glucose (HG100) as well as protein (HG100). As a conclusion PPC as alternative protein in fish nutrition is only feasible with LG contents as HG diets resulted in severe undernourishment of fish, lowest growth performance and irreversible intestinal damages.

α-Tocopherol in weaning diets for European sea bass (Dicentrarchus labrax) improves survival and reduces tissue damage caused by excess dietary DHA contents

The objective of the present study was to investigate the combined effect of several dietary contents of vitamin E and polyunsaturated fatty acids (PUFA), mainly docosahexaenoic acid (DHA), on growth, survival, biochemical composition and tissue morphology of sea bass along early development. A feeding experiment was conducted in sea bass larvae using five different diets with the same proximate composition and different ratios of DHA concentrated fish oil [10, 30 and 50 g kg−1 dry weight (DW)] and vitamin E (α-tocopherol acetate) (1500 and 3000 mg kg−1 DW). DHA was readily deposited in fish tissues and associated with higher sea bass mortalities probably because of increased peroxidation risks. Besides, the elevation of dietary DHA contents up to 5% severely increased the incidence of muscular lesions and the presence of ceroid pigment within hepatocytes. However, elevation of dietary vitamin E levels markedly reduced the incidence of these symptoms in sea bass, increasing the tissue content in several PUFA and improving growth and stress resistance. Moreover, when sea bass was fed diets containing high vitamin E levels, fish showed a significant improvement in growth when dietary DHA was raised from 1% to 3%. Therefore, in sea bass larvae, a ratio of 30 g kg−1 DHA and 3000 mg kg−1 vitamin E seems to be adequate to achieve a good larval performance and to avoid muscular lesions.

Hermansky‐Pudlak Syndrome: Severe Colitis and Good Response to Infliximab

Hermansky‐Pudlak Syndrome: Severe Colitis and Good Response to Infliximab

Multifocal Granulomatous Panniculitis with Ceroid Pigment in Two Mediterranean Striped Dolphins (Stenella coeruleoalba)

Two striped dolphins (Stenella coeruleoalba) were found stranded on the Catalonian Spanish coast. The main pathologic finding in both animals was the existence of multiple granulomatous lesions in the blubber, microscopically composed of macrophages and multinucleated cells containing vacuolar material. This material was identified as ceroid pigment due to its ultrastructural morphology, autofluorescence, and positive staining with periodic acid-Schiff and Ziehl-Neelsen techniques. The special stains and electron microscopy did not reveal any microorganisms associated with the lesions. These findings are very suggestive of "nutritional panniculitis," a well-defined entity associated with vitamin E deficiency that has been rarely described in free-living species.

Albinism, cyclic neutropenia, and ceroid pigment in the liver #

Albinism, cyclic neutropenia, and ceroid pigment in the liver #

Neuronal pigmented autophagic vacuoles: lipofuscin, neuromelanin, and ceroid as macroautophagic responses during aging and disease

The most striking morphologic change in neurons during normal aging is the accumulation of autophagic vacuoles filled with lipofuscin or neuromelanin pigments. These organelles are similar to those containing the ceroid pigments associated with neurologic disorders, particularly in diseases caused by lysosomal dysfunction. The pigments arise from incompletely degraded proteins and lipids principally derived from the breakdown of mitochondria or products of oxidized catecholamines. Pigmented autophagic vacuoles may eventually occupy a major portion of the neuronal cell body volume because of resistance of the pigments to lysosomal degradation and/or inadequate fusion of the vacuoles with lysosomes. Although the formation of autophagic vacuoles via macroautophagy protects the neuron from cellular stress, accumulation of pigmented autophagic vacuoles may eventually interfere with normal degradative pathways and endocytic/secretory tasks such as appropriate response to growth factors.

La fibrose pulmonaire du syndrome de Hermansky Pudlak n’est pas totalement commune

A 55-year-old female patient with Hermansky-Pudlak syndrome (albinism, thrombopathia and ceroid accumulation) underwent a single lung transplantation for pulmonary fibrosis. Examination of explanted lung showed usual interstitial pneumonia pattern associated with two unusual lesions: presence of numerous macrophages containing ceroid pigments within fibrosis and characteristic foamy swelling of pneumocytes. This later lesion, which has only been recently described, seems related to the mechanism of the disease by defect of surfactant secretion. This lesion allows the histological individualization of pulmonary fibrosis associated with Hermansky-Pudlak syndrome from idiopathic usual interstitial pneumonia.

Pigments in aging: an overview.

Although during the normal aging process there are numerous pigmentary changes, the best recognized are those of melanin and lipofuscin. Melanin may increase (e.g., age spots, senile lentigo, or melanosis coli) or decrease (e.g., graying of hair or ocular melanin) with age, while lipofuscin (also called age pigment) always increases with age. In fact, the time-dependent accumulation of lipofuscin in lysosomes of postmitotic cells and some stable cells is the most consistent and phylogenetically constant morphologic change of aging. This pigment displays a typical autofluorescence (Ex: approximately 440; Em: approximately 600 nm), sudanophilia, argyrophilia, PAS positiveness, and acid fastness. Advances on its biogenesis, composition, evolution, and lysosomal degradation have been hampered by the persistent confusion between lipofuscin and the large family of ceroid pigments found in a variety of pathological conditions, as evidenced by the frequent use of the hybrid term lipofuscin/ceroid by investigators mainly working with in vitro systems of disputable relevance to in vivo lipofuscinogenesis. While lipofuscin and ceroid pigments may share some of their physicochemical properties at one moment or another in their evolutions, these pigments have different tissue distribution, rates of accumulation, origin of their precursors, and lectin binding affinities. Although it is widely believed that lipofuscin is a marker of oxidative stress, and that it can be, therefore, modified by antioxidants and prooxidants, these assumptions are mainly based on in vitro experiments and are not generally supported by in vivo studies. Another common misconception is the belief that lipofuscin can be extracted from tissues by lipid solvents and measured spectrofluorometrically. These and other disturbing problems are reviewed and discussed in this presentation.

What is lipofuscin? Defining characteristics and differentiation from other autofluorescent lysosomal storage bodies

Lipofuscins, also known as age-pigments, have three defining characteristics: (1) they consist of intracellular secondary lysosomes; (2) they have a yellow autofluorescent emission when excited by near ultraviolet or blue light; and (3) they accumulate during normal senescence. Lysosomal storage bodies with similar fluorescence properties accumulate in various cell types as a result of specific pathological conditions or experimental manipulations. As a class, the latter are often referred to as ceroid pigments. In general, the mechanisms involved in the formation of ceroid pigments cannot be assumed to be closely similar to those involved in lipofuscin formation. In fact, the mechanisms of formation almost certainly differ, not only between lipofuscins and ceroids, but also among different lipofuscins and different ceroids. Presently, the most detailed knowledge about the mechanisms involved in lipofuscin formation come from studies on the retinal pigment epithelium (RPE) of the eye. These studies indicate that at least the autofluorescent constituents of RPE lipofuscin are generated from derivatives of vitamin A that occur in the retina. Oxidative stress to the retina appears to promote the formation of these RPE fluorophores. Whether similar mechanisms are involved in the formation of the lipofuscins that occur in other tissues remains to be determined. The mechanisms involved in RPE lipofuscin fluorophore formation are closely related to metabolic pathways that are specific to the retina. Thus, it appears likely that the mechanisms by which lipofuscins form in other tissues differ fundamentally from those that underlie RPE lipofuscin formation.

Morphometry of age pigment (lipofuscin) and of ceroid pigment deposits associated with vitamin E deficiency

Consistent amounts of lipofuscin and of ceroid pigment associated with vitamin E deficiency are reported to represent morphological correlates of aging and increased oxidative stress. A reliable quantification of these yellow autofluorescent deposits is of critical biological significance, thus we carried out a computer-assisted morphometric study on the accumulation of lipofuscin in physiological aging and of ceroid pigment in vitamin E deficiency, respectively. The total area and the size distribution of lipofuscin or ceroid pigment deposits were measured in CA3 hippocampal pyramidal neurons of 6-, 12-, 18- and 25-month-old rats, as well as in vitamin E deficient animals of 18 months of age. An increase in the mean total area of lipofuscin and ceroid pigment was found in aging and in vitamin E deficiency. In both conditions, the proportion of large discrete deposits also increased. The similarity of changes observed in old and adult vitamin E deficient animals suggests that the underlying processes initiated by the absence of α-tocopherol from the diet of adult rats and in physiological aging may share some common mechanisms.