Lipofuscin Pigment Research Articles

Allocortical involvement in Huntington's disease.

Seven brains of individuals who had suffered from Huntington's disease and seven control brains were examined. Preparations stained for Nissl material and lipofuscin pigment revealed a characteristic layer-specific loss of nerve cells in two allocortical areas, the entorhinal region and the subiculum. The most severe changes occurred in the entorhinal layer Pri-gamma while layer Pre-alpha, layer Pre-gamma and the subiculum showed less severe alterations.

Intriguing Case: Oncocytic Adrenal Cortical Adenoma

Follow-up examination of a 61-year-old woman who recently had a papillary transitional cell carcinoma removed from her urinary bladder revealed a 5-cm mass in the left adrenal gland. Clinicopathologic studies showed the tumor to be a heretofore undocumented virilizing oncocytic adrenal cortical adenoma. Ultrastructural examination disclosed that the cytoplasm was filled with mitochondria, many of which had tubulovesicular cristae. The smooth endoplasmic reticulum was well developed, as was the Golgi apparatus. Lysosomes of various types, including lipofuscin pigment, were also evident. Immunohistochemical studies revealed variable immunoreactivity for simple epithelial cell cytokeratins and for synaptophysin (weak staining) in a minority of tumor cells. Two additional cases of oncocytic adrenocortical tumors, presumably adenomas, from other institutions are also discussed. The small size and weight of the tumor, the low mitotic rate, and the absence of necrosis and vascular and capsular invasion portend a good prognosis. The patient is currently free of disease 1 year after extirpation of the tumor. Additional cases and longer follow-up times are needed to predict better the behavior of this rare oncocytic adrenal cortical neoplasm.

Transmural differences of lipofuscin pigment accumulation in the left ventricle of rat heart during growth and aging1

In view of the higher metabolic rate in subendocardial heart tissue, the rate of age-related lipofuscin pigment accumulation was explored in different regions of the left ventricle heart wall of Sprague-Dawley rats. Hearts were removed from 2-, 6-, 12- and 24-month-old rats, and lipofuscin pigment accumulation was assessed in the subepicardial and subendocardial layers, either by measuring extractable fluorescent material, or by direct visualization with fluorescence microscopy. Findings showed that the amount of extractable fluorescent material and the number, size and brightness of the fluorescent lipofuscin granules increased with age in all the myocardial tissue layers. The rate of accumulation of extractable fluorescent material was higher in subendocardial compared to subepicardial tissue. At the microscope, fluorescent granules exhibited a different morphological appearance in the subendocardial and subepicardial tissue of the two older age-groups. These data support the hypothesis that liposoluble age-pigment deposition is linked to the rate of local oxidative metabolism.

Hyaline Droplet Accumulation in Rodent Kidney Proximal Tubules: An Association with Histiocytic Sarcoma

Since recognition during the last decade that certain renal carcinogens can initially cause an accumulation of hyaline (protein) droplets in proximal tubules of male rats, it has become appropriate to establish whether this phenomenon of protein overload can also occur in rodent kidneys unrelated to chemical treatment. Kidney tissue from a number of selected rodent studies held in the National Toxicology Program (NTP) or Food and Drug Administration (FDA) archives were evaluated for hyaline droplet accumulation in proximal tubules. The survey concentrated on rats and mice of both sexes bearing hematopoietic tumors, as our preliminary observations had suggested this direction of study. The tissues of 101 Sprague-Dawley, 25 Osborne-Mendel, and 70 Fischer 344 rats and 96 B6C3F1 mice were examined. These animals provided an assortment of tumors including histiocytic sarcoma, lymphocytic lymphoma, mononuclear cell leukemia, and sarcoma. Hyaline droplet accumulation, primarily involving the P2 segment of proximal tubules, was diagnosed in 96% of rats with histiocytic sarcoma (74/77 cases in Sprague-Dawley, 17/18 in Osborne-Mendels, 7/7 in Fischers) and in 55% of B6C3F1 mice with histiocytic sarcoma (18/33 cases). There appeared to be a qualitative correlation between hyaline droplet accumulation and degree of tumor burden. Thus, in cases negative for hyaline droplets, the tumor was often confined to a single location, while increasing involvement of proximal segments beyond P2 occurred with more extensive multi-organ dissemination of the tumor. By immunohistochemistry on 11 cases of rat and 8 cases of mouse histiocytic sarcoma, the protein in hyaline droplets was identified as lysozyme, a known major secretory product of monocytes and macrophages. The hyaline droplets were negative for alpha 1-antitrypsin, alpha 2u-globulin, rat or mouse immunoglobulin, and albumin. More sparsely scattered droplets and granules present in proximal tubules of Fischer rats with mononuclear cell leukemia were negative for lysozyme but positive for either iron or lipofuscin pigment. The study establishes a clear association between renal tubule hyaline droplet and lysozyme accumulation in rats and mice with histiocytic sarcoma. Hyaline droplets secondary to neoplasia should be distinguished from chemically-induced hyaline droplet nephropathy in the male rat involving alpha 2u-globulin.

Endrin-Induced Histopathological Changes and Lipid Peroxidation in Livers and Kidneys of Rats, Mice, Guinea Pigs and Hamsters

Endrin toxicity may be due to an oxidative stress associated with increased lipid peroxidation, decreased glutathione content, and inhibition of glutathione peroxidase activity. Extensive interspecies variability exists in sensitivity towards endrin. Therefore, histopathological changes and lipid peroxidation in the livers and kidneys of rats, mice, hamsters, and guinea pigs were examined 24 hr after the administration of 4 mg endrin/kg body weight orally in corn oil. Degeneration and necrotic changes with inflammatory cell infiltration were observed in livers and kidneys, and interspecies variability occurred. Fatty changes in the form of hepatic foam cells with cytoplasmic vacuolation were present. Lipofuscin pigments, associated with lipid peroxidation, were observed in hepatocytes and Kupffer cells. These histopathological conditions were prevented in rats which had been pretreated with butylated hydroxyanisole, vitamins E and C, or cysteine, antioxidants and free radical scavengers which have previously been shown to inhibit lipid peroxidation. The extent of endrin-induced lipid peroxidation correlated well with the degree of histopathological changes. Thus, histological changes consistent with the induction of an oxidative stress were observed following the administration of endrin to various animal species.

Glutamic acid decarboxylase (GAD)-immunoreactive structures in the adult human lateral geniculate nucleus

The distribution of glutamic acid decarboxylase (GAD)-like immunoreactivity in the lateral geniculate nucleus (LGN) of the human adult was studied in vibratome sections (50-60 microns thick) using the avidinbiotin-peroxidase method. The tissue was obtained at autopsy from five individuals without any known neurological disorders. Only few GAD-immunoreactive neurons were present in the layers of the LGN, even less in the interlaminar zones. The numerical density of GAD-immunoreactive neurons and puncta (probably synaptic boutons and or cross sectioned cell processes) in the magnocellular layers was larger than in the parvocellular layers. Furthermore, no striking differences between the individual parvocellular layers were noted. The immunoreactive somata were polygonal or triangular, occasionally pear-shaped, and ranged in size from 15 to 25 microns. They gave off two to four short, thick, straight primary dendrites. A preferred orientation of dendrites was not recognized. After bleaching the chromogen 4-chloro-1-naphthol and staining for lipofuscin pigment granules and basophilic material, 254 unequivocally relocated GAD-immunoreactive nerve cells could be classified as belonging to the lipofuscin pigment granules-containing class of interneurons.

Lipid pigment (lipofuscin) accumulation in rat tongue muscle with aging

Lipid pigment (lipofuscin) was examined in perioral and tongue muscles of aging rats. While in perioral striated muscle the pigmented granules could not be found, a considerable accumulation with aging could be noted in the tongue muscle. Unlike the human tongue muscle, in which the pigmented granules were stored in clusters of different sizes at the nuclear poles of the muscle fibers, in rats, they were scattered randomly through the cytoplasm. The results of this study suggest that lipofuscin can be used as a marker for aging of the tongue muscle in the rat.

Oxidants and antioxidants in aquatic animals

1. 1. Oxidative stress, potentially, is experienced by all aerobic life when antioxidant defenses are overcome by prooxidant forces, and is the basis of many physiological abberations. 2. 2. Environmental contaminants may enhance oxidative stress in aquatic organisms, e.g. highly elevated rates of ideopathic lesions and neoplasia among fish inhabiting polluted environments is increasingly related to oxidative stress associated with environmental pollution. 3. 3. Metabolism of redox cycling xenobiotics in aquatic organisms is very similar to that of mammals suggesting similarities in the health consequences of exposure to such compounds. 4. 4. The expression of specific lesions known to arise specifically from oxidative stress, e.g. lipid peroxidation, oxidized bases in DNA and accumulation of lipofuscin pigments are present in many aquatic animals exposed to contaminants. 5. 5. Aquatic organisms contain the major antioxidant enzymes SOD, catalase and glutathione peroxidase, albeit there are marked quantitative differences among the various species reported.

Morphological correlates of aging in the chinchilla cochlea

The inner ears from 80 chinchillas ranging in age from premature to 19.2 years were examined as plastic-embedded flat preparations to determine the morphological changes associated with aging. Three of the four forms of human presbycusis defined by Schuknecht were found in the chinchillas. All animals had losses of sensory cells or sensory presbycusis. Inner (IHCs) and outer hair cells (OHCs) degenerated at a rate of about 0.29% and 1.0% per year, respectively. Age-related degeneration of inner (IPs) and outer pillars (OPs) occurred at a much slower rate. In four animals (5%) the dendritic processes of some of the spiral ganglion cells had degenerated in areas where the loss of sensory cells was minimal. This pathological change is likely equivalent to neural presbycusis. Six animals (7.5%) had regions of degeneration of the stria vascularis or striai presbycusis. The other common finding in the aging cochleas was the presence of lipofuscin or age pigment. Lipofuscin deposits were found to accumulate in the subcuticular region of OHCs, IPs and OPs, near the endolymphatic surfaces of many of the supporting cells and in the epithelial cells of Reissner's membrane. The IHCs accumulated much less lipofuscin. The morphological changes seen in the ears of aging chinchillas were qualitatively similar to those seen in the temporal bones of aging humans although the magnitude of the changes was considerably less. These results suggest that some of the damage found in aging human cochleas may be due to aging plus exposure to one or more ototraumatic agents.

A quantitative and histochemical study on melano‐macrophage centres in the spleen of the teleost fish Diplodus annularis L.

The incidence and histochemical features of melano‐macrophage centres in the spleen of the teleost fish D. annularis were studied with respect to starved and fed fish. Tissue catabolism following complete starvation appears to induce formation of new centres containing melanin and lipofuscin pigments. However, a change in the amount of melano‐macrophages was also recorded in fed fish, suggesting that some other factor(s), internal to fish, may affect the presence of such structures within the spleen of D. annularis.

Skin pigmentation from clofazimine therapy in leprosy patients: A reappraisal

Skin biopsy specimens from two lepromatous leprosy patients with dark brown pigmentation who were receiving long-term clofazimine therapy were studied. Ceroid-lipofuscin pigment was demonstrated inside macrophages that contained numerous phagolysosomes. These contained lipids and clofazimine that appeared as electron-lucent vacuoles and a lipofuscin pigment that was electron dense, granular, and lamellated. Although the presence of the drug in tissues contributed to the skin pigmentation, the main cause was a drug-induced, reversible ceroid lipofuscinosis.

Neuroaxonal dystrophy associated with vitamin E deficiency in two Haflinger horses

Neuroaxonal dystrophy and minimal diffuse degenerative myelopathy was diagnosed in two female full sibling, 1- and 2-year-old, Haflinger horses. Both animals developed slowly progressive ataxia from the age of 4 months. Clinical signs, which were more prominent in the hind legs, included hypermetria and dysmetria. Histological examination revealed neuroaxonal dystrophy characterized by spheroid formation, vacuolation, astrogliosis and lipofuscin pigment deposition in macrophages and neuronal perikarya. These changes were restricted to the gracilis and cuneate nuclei, nucleus of the solitary tract, nucleus intermediomedialis in the sacral and the seventh segment of the cervical spinal cord and Stilling-Clarke's column in both horses and the medial vestibular nucleus in the older horse. Both diseased Haflingers had significantly reduced serum alpha- and gamma-tocopherol values. No significant alteration in serum total lipid concentrations and serum selenium values were observed. It is likely that the condition has a familial hereditary basis. It is unclear whether there is a link between the observed neuroaxonal dystrophy and vitamin E deficiency and further investigations are warranted.

Lipofuscin pigments in the spiral ganglion of the rat

The purpose of the present study was to clarify morphological differences in lipofuscin or so-called age pigments observed in the spiral ganglion cells of both young and adult rat groups and to characterize the size and structure of ceroid pigment granules generated in vitamin-E-deficient rats. The results showed different patterns of lipofuscin distribution in the two groups. The adult rat group had large aggregated lipoid, dark pigment granules of irregular shape in the cytoplasm. In contrast, the young group had small numbers of small, dense homogeneous granules, suggesting higher Schwann cell phagocytic activity. The ceroid pigments apparently included numerous vesicles and droplets of more variable density and size than the lipofuscin pigments appearing in the non-treated older animals. Both lipofuscin and ceroid pigments developing in such non-dividing cells are produced as a result of peroxidation reactions, so that the more they accumulate in the cytoplasm the more likely cell function deteriorates. The present study has shown that lipofuscin/ceroid granules are generated in the spiral ganglions under either endogenous (aging) or exogenous (vitamin E deficiency) conditions.

β-Protein immunoreactivity in brains of patients with neuronal ceroid lipofuscinosis: Ultrastructural and biochemical demonstration

The storage pigment in neuronal ceroid lipofuscinoses (NCL) has a close similarity to age pigment lipofuscin. We studied immunoreactivity of isolated neuronal pigments from the juvenile form of NCL and aging control, using monoclonal antibodies (mAbs) against amyloid β-protein. Ultrastructural localization of the immunoreactivity demonstrated that in NCL the epitopes are distributed mainly in curvilinear multilamellar arrays of the storage pigments and less in fingerprint profiles, while in aging control they are more homogeneously distributed on age pigment lipofuscin. The different distribution of the epitopes may reflect some catabolic as well as morphologic differences in lysosomes. A unique 31-kDa polypeptide detected on Western blots in NCL possibly derives from the same precursor, amyloid β-protein precursor (ABPP). ABPP processing may be aberrant in NCL brains, and this can be detected as a 31-kDa polypeptide reactive with the mAbs.

Leydig cell cytoplasmic content is related to daily sperm production in men.

The relationship between the abundance of specific Leydig cell organelles and daily sperm production (DSP) was determined. Testes from 10 men (26-53 years of age) were obtained at autopsy within 10 h of traumatic death or heart failure and fixed by vascular perfusion. Testicular tissue was processed for light and electron microscopy. DSP/testis and Leydig cell cytoplasmic volume/testis were determined by stereology of histologic sections. The Leydig cell organelle content was determined by point counting electron micrographs for smooth endoplasmic reticulum (SER), rough endoplasmic reticulum, mitochondria, lipofuscin pigment, lipid, Golgi bodies, and Reinke crystals. Men were divided equally into two groups based on DSP/testis. Men with low DSP/testis had less SER volume density (P less than 0.01) and lower SER volume per testis (P less than 0.05) than men with high DSP. Other organelles were unrelated to DSP. When all men were combined, the volume density of SER (r = 0.80; P less than 0.01), the volume SER per testis (r = 0.69; P less than 0.05), and the volume SER per Leydig cell (r = 0.84; P less than 0.01) were significantly related to DSP. Hence, there appears to be a significant relationship between Leydig cell SER and the level of spermatogenesis in men.

Correlated autoradiographic and ion-microscopic study of the role of iodine in the formation of ?cold? follicles in young and old mice

The role of iodine in the formation of "cold" follicles (not labeled on autoradiograms after radioiodine administration) was analysed in ICR female mice during aging and involution of thyroid hyperplasia, by use of light and electron microscopy and by comparing autoradiographic and analytical ion-microscopic images for the same follicle in serial sections. The proportion of "cold" and "partly cold" (displaying a patchy or ring labeling pattern on autoradiograms) follicles increased significantly during aging. This increase was more pronounced in old mice fed an iodine-rich diet as compared to mice fed a moderate iodine diet. Similarly, during goiter involution produced by refeeding iodine, the follicular heterogeneity of iodine metabolism was more accentuated with a high dose of iodine, regardless of the age of the mice. The follicular lumina of "hot" and "cold" follicles had the same concentration of stable iodine, as shown by analytical ion microscopy, and the cells of both types of follicles formed colloid droplets in response to TSH. Furthermore, when a goitrogenic treatment was induced in aged mice, some "cold" follicles persisted after 8 days, but all follicles resumed "hot" after 16 days. By analytical ion microscopy, 127iodine was also found inside thyroid cells of old mice, but the cytoplasmic patches of 127iodine were not labeled with 125iodine. They corresponded to lipofuscin pigments and secondary lysosomes, as observed in serial sections at the electron-microscopic level. This intracellular stable iodine could constitute a slow turnover compartment not used for hormone synthesis.

Comparative toxicology of temelastine: A novel H 1 antagonist in dog, rat, and monkey

The toxicity of temelastine 2-[4-(5-bromo-3-methylpyrid-2-yl)butylamino]-5-[(6-methylpyrid-3-yl) methyl]-4-pyrimidone a potent, selective, competitive histamine H 1-receptor antagonist was examined in dogs and rats. The major toxicological response seen in the dog was marked, but intermittent and reversible, increases in the plasma activity of a number of liver-associated enzymes, viz alanine aminotransferase (ALT), glutamate dehydrogenase (GLDH), and alkaline phosphatase (ALP). The increases first seen in two male dogs treated for 30 consecutive days at a dose of 300 mg/kg became apparent at lower doses, i.e., 100 and 33.3 mg/kg/day, in 6- and 12-month studies. Although the increases were suggestive of hepatotoxicity, the only histological changes were increases in hepatocellular lipofuscin pigment and foci of macrophages seen in dogs treated at 300 mg/kg for 12 months. Rats treated for up to 12 months at doses as high as 300 mg/kg/day showed no treatment-related increases in plasma enzymes although increases in liver weights and hepatocellular lipofuscin pigment together with centrilobular hypertrophy were seen in the 300 mg/kg/day treatment group. To investigate differences in hepatic responsiveness between species dogs, rats, and monkeys were exposed to high concentrations of temelastine by continuous 24-hr intravenous infusion. The results of the study showed the dog to be most sensitive to the hepatic effects of temelastine. The major toxicological effect of temelastine in the rat was a histopathological lesion of the thyroid gland characterized by agglomeration and depletion of colloid, follicular epithelial hypertrophy and reduced follicular size. The no-effect dose for this lesion was between 10 and 33.3 mg/kg/day. These histopathological changes, characteristic of a “TSH-driven” thyroid gland, were not seen in the thyroid glands of dogs.

Glutamic‐acid‐decarboxylase‐ and parvalbumin‐like‐immunoreactive structures in the olfactory bulb of the human adult

This study examines the distribution and morphological characteristics of glutamic-acid-decarboxylase-like (GAD)- and parvalbumin-like (PA)-immunoreactive structures in the olfactory bulb of the human adult. GAD-immunoreactive somata occurred in the glomerular layer, the external granule cell layer, the more superficial portion of the external plexiform layer, and the internal granule cell layer. The cells were small- to medium-sized. Demonstration of lipofuscin pigment revealed the presence of unpigmented as well as pigmented neurons, thus suggesting the existence of two subpopulations of GAD-positive neurons. GAD-immunoreactive puncta and/or fibers were mainly seen in the periglomerular region and the internal granule cell layer. All other layers of the bulb, as well as the intrabulbar portion of the anterior olfactory nucleus, displayed considerably less of these puncta and/or fibers. The olfactory nerve layer remained practically clear of immunoreactive material. PA-immunoreactive somata occurred in the glomerular layer and both the external and internal granule cell layer. Only a small number of immunoreactive nerve cells were encountered within the white matter or the olfactory tract. Most PA-positive neurons displayed characteristics of short axon cells whereas a few others resembled van Gehuchten cells. All of the PA-immunoreactive neurons were devoid of lipofuscin pigment. Immunoreactive puncta and fibers were present in all layers though predominating in the periglomerular region, the olfactory nerve layer, and the internal granule cell layer. The intrabulbar portions of the anterior olfactory nucleus did not show any immunoreactive structures.

Hepatocellular Lipofuscin in Pre‐Eclampsia

Thirty liver specimens from 30 pre-eclamptic women with and without liver dysfunction were examined for the amount of lipofuscin pigment deposited. The amount of hepatic lipofuscin correlated positively with plasma urate concentrations. This finding may indicate an involvement of oxygen-free radicals and xanthine oxidase in the pathophysiology of pre-eclampsia.

Altered lipofuscin pigmentation in the basal nucleus (Meynert) in Parkinson's disease.

The basal nucleus (Meynert) and the horizontal limb nucleus of the diagonal band are composed of 3 types of neurons that can be differentiated with the use of Nissl preparations counterstained for the demonstration of lipofuscin pigment. The pattern of pigmentation of these neurons has been investigated in parkinsonian cases and compared with controls. In the parkinsonian cases the number of large multipolar neurons showing pigment granules in a dendritic stem is significantly higher than in controls. Moreover, the degree of pigment penetration is more pronounced in parkinsonian cases. The results show that staining of lipofuscin granules offers an approach to detecting further details of the changes in the basal nucleus in Parkinson's disease.