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Abstract
A study was made of the effects of centrophenoxine on the learning and memory of old mice. The results were correlated with changes in neuronal lipofuscin in the cerebral cortex and hippocampus. Old female mice (11-12 months) were treated with centropheoxine for three months and their learning and memory were tested in a T-maze. The number of trials required to attain the criterion in the 20 treated old mice were compared with those for 20 untreated mice of the same age and for 20 younger untreated mice. The treated animals learned the task with significantly fewer trials, and also exhibited a reduction of neuronal lipofuscin pigment in both the cerebral cortex and the hippocampus. The changes in lipofuscin were demonstrated by study of the characteristic autofluorescence, and by histolchemical and ultrastructural (electron microscope) observations.
... Meclophenoxate is usually considered an aging reversal drug, showing its neuroprotective effect in the brain and even prolonging the life span (Hochschild 1973). It positively influenced learning and memory in old mice, and the effect correlated with the reduction of neuronal lipofuscin in the cerebral cortex and hippocampus (Nandy 1978). The primary mechanism of meclophenoxate action is Fig. 4 Chemical formula of meclophenoxat generally believed to be cholinergic. ...
... Centrophenoxine (CPH) is an ester of dimethyl-aminoethanol (DMAE), a component in choline synthesis, and p-chlorophenoxyacetic acid, a synthetic form of a plant growth hormone. 41 Elevation of brain acetyl choline is one mechanism for therapeutic effects in cerebral atrophy, dementia, and TBI. CPH also delivers DMAE rapidly to the brain where it is incorporated into nerve cell membranes as phosphatidyl-DMAE, an avid scavenger of OH-radicals. ...
Herbal medicines supported by evidence of safety and efficacy in the treatment of anxiety, insomnia, fatigue, cognitive enhancement, mental focus, and sexual function are useful as monotherapies, multiherb combinations, and as adjuncts to prescription psychotropics. Relevant mechanisms of action and clinical guidelines for herbs in common use can assist clinicians who want to enhance treatment outcomes by integrating phytomedicinals into their treatment regimens. Research is needed to strengthen the evidence base and to expand the range of disorders that can be treated with herbal extracts. Studies of herbal genomic effects may lead to more targeted and effective treatments.
... The decrease in amylase activity in salivary glands, particularly parotid and submandibular glands, can be explained from several perspectives, but mainly there may be change in fidelity of transcription and translation, which are changes in response to intrinsic as well as extrinsic factors. Centrophenoxine is one of the potent antioxidants that reduce the peroxides formed in the tissues during aging (Nandy, 1978). Thus, it is possible that the antioxidant centrophenoxine is efficient in reducing peroxidation in salivary glands. ...
Three month old female mice were ovariectomised and after 4 days the mice were separated into six groups, I) ovariectomized, II) ovariectomized + D-galactose treated, III) ovariectomized + D-galactose treated + Tween 80, IV) ovariectomized + D-galactose & centrophenoxine treated, V) ovariectomized D-galactose treated + olive oil injected, and VI) ovariectomized + D-galactose treated + estrogen injected. After 15 days the mice were sacri-ficed. The protein concentration, amylase activity, lipid peroxidation and fluorescence product were measured in various salivary glands, viz., submandibular, sublingual, and parotid. In ovariectomized (group II) and ovariec-tomized + D-galactose treated (group III) mice the protein concentration and amylase activity of various salivary glands decreased, while lipid peroxidation and fluorescence product increased. In ovariectomized + D-galactose + centrophenoxine treated (group IV) and ovariectomized + D-galactose treated + estrogen injected (group VI) mice protein concentration and amylase activity in the salivary glands increased, while lipid peroxidation and fluorescence product decreased. These results reveal that centrophenoxine, and therefore aging, has modulatory effects on salivary glands in D-galactose induced aged female mice.
Nootropics are agents enhancing cognitive functions without causing sedation or stimulation. Several different pharmaceuticals have been identified and evaluated for their possible effects in treating cerebral insufficiency, and piracetam was the first representative of this group of medications. Current evidence from animal studies on such drugs as piracetam, pyritinol, co-dergocrine, meclophenoxat, pentoxifylline, and nimodipine therapeutic applications is convincing. They have been shown to improve cerebral blood flow and influence neurotransmission and receptors essential for cognitive functioning and antioxidant activity. Considering a relatively good safety profile, they could be recognized as an adjuvant to any standard therapy for cognitive disorders, where current therapeutic options are limited. Unfortunately, despite several interesting trends, clinical data are somewhat limited, mixed, contradictory, or inconclusive. The published studies are mostly old, and some are of suboptimal quality. They included a relatively small number of participants, and the duration of therapy was most often comparatively short. Old criteria for determining cognitive impairment and dementia assessment instruments insufficiently sensitive to evaluate cognition changes over time were used. Lack of validated clinical trial measures designed to assess prodromal dementia was noticed. Moreover, considerable variation in research methodology preventing their joint analysis was observed. Therefore, the overall usefulness of these pharmaceuticals in various central nervous system disorders as a supplement or adjuvant therapy needs to be established through high-quality multicenter randomized controlled clinical trials with sufficient sample size and optimized study design before their widespread use can be recommended.
The single most dramatic change in the United States population age distribution during this century has been the increasing proportion of the population 65 years and over. It has increased from 4.1% in 1900 to 9.9% in 1970, and it will increase to at least 10.6% by the year 2020. In absolute numbers, it has increased from 3.2 million in 1900 to 20.2 million in 1970. The projected increase is 40.2 million by the year 2025 (Cutler and Harootyan, 1975). The most specific and effective way to show this change in the proportion of the elderly is to use an age-sex population “pyramid” (Cutler and Harootyan, 19 75; Domino et al., 1978). Figure 1 depicts graphically the projected dramatic change in composition of the total Unites States population by age and sex from 1975 to 2025.
Wenn Biologen etwas zu dem hier zu diskutierenden Thema beitragen, so liegen deren Schwierigkeiten sicherlich nicht an der erst kürzlich eingeführten Terminologie, sondern eher darin, daß Biologen nicht das vermitteln bzw. nicht vermitteln können, was aus der Sicht der Medizin von ihnen erwartet wird.
The term ‘cerebral activating drugs’ is used for the title of this chapter as a convenient way to embrace a wide variety of drugs which have been used in attempts to improve cerebral function in old age. Such drugs include vasodilators, drugs influencing cerebral metabolism, stimulants and drugs which are claimed to improve the flow of blood in the cerebral microcirculation by virtue of actions on blood viscosity, red cell deformability, or one or more biochemical effects at the cellular level.
Die Entwicklung der Gerontopsychopharmakologie wurde nicht, wie das für die allgemeine Psychopharmakologie der Fall war, durch die zufällige Entdekkung klinisch eindeutig wirksamer Substanzen — wie z. B. des antipsychotisch wirksamen Chlorpromazins im Jahre 1952 durch Delay u. Denniker — eingeleitet. Hinzukommt, daß die systematische wissenschaftliche Untersuchung dieser Substanzen ihrerseits eine auf pharmakologischen Hinweisen beruhende biologische Ursachenforschung der Depression und Schizophrenie induziert hat. Da ähnliche „glückliche“ Entdeckungen bisher der Gerontopsychopharmakologie nicht zuteil wurden, sind in diesem Bereich andere Strategien zur Entwicklung einer rationalen Gerontopharmakotherapie erforderlich. Eine sich anbietende Chance liegt vor allem in der Intensivierung der Erforschung der biochemischen Grundlagen der morphologischen Veränderungen des sog. physiologischen Alterungsprozesses sowie der degenerativ bzw. vaskulär bedingten Abbauerkrankungen. Das Fehlen pharmakologischer Wegweiser im oben genannten Sinne muß dabei durch die auf der Basis der Grundlagenforschung entwikkelten hypothetischen pharmakologischen Modelle ersetzt werden. Wenn auch bisher die Mehrzahl der zu prüfenden hypothetischen pharmakologischen Modelle im Endergebnis nicht bestätigt werden konnten, so liegt ihr Wert trotzdem in der Etablierung einer beginnenden und naturgemäß noch sehr simplifizierenden rationalen gerontopharmakologischen Forschung. Diese Forschungsansätze einer rationalen Gerontopharmakologie sollten in Zukunft stärker gefördert und unterstützt werden.
This paper presents experimental data regarding the synthesis and the antimicrobial activity of new 4-phenylazo-phenoxyacetic acids. All compounds were characterized by IR, UV-Vis, mass spectral data and by elemental analysis. These compounds were tested for their antibacterial activity against Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris and for their antifungal activity against Candida albicans by disk diffusion method.
The term “lipofuscin” was introduced in the literature by Borst (1922), although the pigment was first demonstrated by Hannover (1842). The name was derived from a Greek word lipo (meaning fat) and a latin word fuscus (meaning brown). The deposition of the pigment in the cytoplasm of the neurons is one of the consistent age-related cytological changes, although it is found in myocardium and skeletal muscle as well. Various investigators demonstrated the pigment and used different names, such as chromolipid, age pigment, and ceroid (Bondareff 1957; Duncan et al. 1960; Pearse 1964; Samorajski et al. 1964). The genesis of lipofuscin has been investigated by a number of workers, but the results are inconclusive (DeDuve 1963; Essner and Novikoff 1960; Friede 1962; Goldfischer et al. 1966; Barka and Anderson 1963; Nandy 1968; Sulkin 1953; Whiteford and Getty 1966). In this chapter, an attempt is made to discuss the pertinent literature on lipofuscin and its possible significance.
The dynamics of the level of direct current potentials and the content of lipid hydroperoxides, conjugated dienes, Schiff bases of phospholipids and lipofuscin in the brain of rats during ontogenesis was studied. It is shown that changes in neurophysiological and biochemical characteristics have an M-shaped character. The synchronization of age-dependent changes in the content of lipid peroxidation products and the dynamics of direct current potentials was revealed.
Among the three main groups of biological macromolecules uniquely linked by transfer and expression of genetic information: (DNA - RNA - Proteins), the RNA stage is the least studied for possible age-related changes. There have been many recent reviews about age-related changes of DNA and proteins, or DNA- protein complexes in chromatin. Recently however, there has been only one review on the role of RNA and RNA metabolism in aging (Rothstein 1982), and a brief supplement to it (Eichhorn 1983) which summarizes the available experimental data up to 1980 on the age-related changes of the synthesis of different types of RNA (rRNA, mRNA, and tRNA). The authors have not linked factual information with several relevant theories of aging [genetic program theory, codon-restriction theory, loss of rRNA genes theory, loss of gene repression theory (dysdifferentiation), general error theory, and others which cannot be proved without special study of transcription and RNA] simply because the information about the age-related changes of RNA is very limited and too general. There have recently been two reviews on the age-related changes of chromatin (Thakur 1984; Medvedev 1984), which also included attempts to cover the connected problems of age-related changes of DNA and the pattern of transcription. Here we would like to select the role of RNA in the aging process for more detailed analysis. The amount of knowledge in this field has not increased dramatically during the last 3–4 years, although there have been many quite interesting studies of changes of RNA in aging tissues which could be added to the list of studies covered by previous reviews.
This chapter discusses lipofuscin and its relevance to the process of aging in an organism. Lipofuscin can be defined as a yellowish brown, auto fluorescent, and lipidcontaining pigment that accumulates in the cytoplasm of cells during aging. It is widely distributed in nature. In man, lipofuscin exists in nearly every part of the body, provided the body is old. The largest accumulations occur in postmitotic cells such as nerve cells and myocardium or in slowly dividing cells such as hepatocytes and adrenal cortical cells. Under the light microscope, lipofuscin consists of light yellow to brownish round or oval granules, measuring about 0.5 to 3 μm in diameter. The younger the individual, the paler, smaller, and sparser the granules tend to be. With advancing years, their numbers increase, they enlarge, and their color becomes darker and more pronounced. The light yellow is known as lipofuscin and the dark brown melanin pigments in nerve cells. Neuromelanin, found in autonomie ganglia and in the pigmented nuclei of the brain stem, consists of granules that are darker and coarser than those of lipofuscin though it behaves much more like lipofuscin than does the melanin of skin and choroid. It is believed that various degenerated cytoplasmic constituents and metabolic by-products contribute to its building blocks, that the completion of the process requires an appropriate balance of intracellular oxidants and antioxidants, and that the final storage is in lysosomes. The role of lipofuscin in the cell, once formed, is unknown, but future research in this field should shed some light on this aspect.
Although difficult to find traces, drugs are believed to have effect of memory and learning. Drug-induced changes in acquisition or extinction are interpreted to indicate that the drug has affected memory processes. However, the effectiveness of drugs that impair (retrograde amnesia) or facilitate (delay extinction) retention is sensitive to dose, type and degree of training, and time of drug administration. The recent trend has been to combine the behavioral results with biochemical measurements in an attempt to identify brain structures and systems that may be involved in the memory process. Many studies that have focused on measuring the consequences of electrical stimulation or microinjection of drugs into specific brain loci have shown that behaviorally-active drugs affect neurotransmitter systems, particularly noradrenergic and dopaminergic pathways. Studies in humans tend to corroborate the data in rodents. For example, administration of scopolamine to young students impaired memory storage and retrieval, as well as other cognitive functions. The impairment of cognitive function in both humans and primates was similar to that found in normal aged subjects. It has been shown that rats that are deficient in vasopressin have deficits in acquiring and maintaining avoidance behavior that can be corrected by treatment with this peptide. Interactions between opiate peptide systems and vasopressin pathways may also be of functional significance in memory processes. Experiments performed using subhuman species suggest that cholinomirnetic drugs should improve memory and cognition in humans who do not perform well, and this view is supported by many favorable clinical reports.
Public and clinician interest in complementary and alternative medicine (CAM) has grown in recent decades, and has been driven in part by concerns about medication side effects, nonresponse to standard medications, and the high cost of prescription medications. CAM (natural products and therapies) for many people already plays a central role in the treatment of psychiatric disorders. As an area of special interest within psychiatry, the concept of integrative psychiatry (IP) supports the development of a treatment approach that more fully incorporates evidence-based use of medicinal herbs, nutrients, and mind–body practices with pharmacotherapy and psychological therapies.
The Sequential Treatment Alternatives to Relieve Depression (STAR*D) trial highlighted the benefits and limitations of combining or switching antidepressants (Rush et al., 2006). Treatment resistance is an indication of our incomplete understanding of the neurobiological basis of mood disorders. For example, paradigms for treating affective disorders are expanding beyond monoamine neurotransmission models to incorporate other regulatory systems (Rizvi & Kennedy, 2011). The increase in information about the effects of immune function, autonomic nervous system balance, neuroendocrine systems, antioxidants (cellular defense and repair), mitochondrial energy transport, the arachidonic acid cascade, second messengers, and gene activation opens the way for new therapeutic approaches, including the use of CAM treatments that may modulate a broad array of the neuropathological underpinnings of psychiatric disorders.
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As was shown in a recent review by this author (Ann. N.Y. Acad. Sci., 928:187-199, 2001), oxyradicals cannot be considered only as harmful by-products of the oxidative metabolism, but living cells and organisms implicitly require their production. This idea is supported by numerous facts and arguments, the most important of which is that the complete inhibition of the oxyradical production by KCN (or by any block of respiration) kills the living organisms long before the energy reserves would be exhausted. This new theoretical approach not only helps our understanding of the normal functions of the living organisms, such as the basic memory mechanisms in the brain cells, but also helps in identifying the site-specific, radical-induced damaging mechanisms that represent the undesirable side effects of oxygen free radicals. First of all, these effects make the cell plasma membrane vulnerable and cause a series of intracellular functional disorders, as described by the membrane hypothesis of aging (MHA). The logical way for any antiaging intervention therefore should be to increase the available number of loosely bound electrons inside the plasma membrane that are easily accessible for OH• free radical scavenging. The present review summarizes the available knowledge regarding the theory of the use of membrane-related antiaging pharmaca, like centrophenoxine (CPH), tested in both animal experiments and human clinical trials. A modified, developed version of CPH coded as BCE-001 is also reported.
This chapter is intended to outline the main results of a research trend realized by the author during the last 45 years, focused on the main role played by the cell membrane in the aging process. It is a very wide field; therefore, the reader cannot expect in this limited space a detailed description, but will be given a wide, interdisciplinary insight into the main facts and theories regarding cellular aging. The central idea described here is the concept called the membrane hypothesis of aging (MHA). The history, the chemical roots, physicochemical facts, biophysical processes, as well as the obligatory biochemical consequences are all touched in by indicating the most important sources of detailed knowledge for those who are more interested in the basic biology of the aging process. This chapter covers also the available anti-aging interventions on the cell membrane by means of the centrophenoxine treatment based on the MHA. It also briefly interprets the possibilities of a just developing anti-aging method by using the recombinant human growth hormone, essential basis of which is the species specificity, and the general presence of receptors of this hormone in the plasma membrane of all types of cells. © 2014 S. Karger AG, Basel.
This author has shown that oxyradicals cannot be considered only as harmful by-products of the oxidative metabolism. Namely, the living state of cells and organisms implicitly requires the production of oxyradicals. Such a concept agrees with many facts and arguments. The most important of them is that the complete inhibition of the oxyradical production by KCN (or by any block of respiration) kills the living organisms much before the energy reserves would be exhausted. This theoretical approach not only helps our understanding of the normal functions of the living organisms, but also helps in identifying the site-specific, radical-induced damaging mechanisms, which represent the non-desired side-effects of the oxygen free radicals. These effects make vulnerable first of all the cell plasma membrane and cause a series of intracellular functional disorders as described by the membrane hypothesis of aging (MHA). The logical way of any anti-aging intervention should be, therefore, to increase the available number of loosely bound electrons inside the plasma membrane, which are easily accessible for OH• free radical scavenging process. The present paper summarizes the available know-ledge regarding the theory of the use of membrane-related anti-aging pharmaca, like centrophenoxine (CPH), tested in both animal experiments and human clinical trials. A modified, further developed version of CPH coded as BCE-001 is also reported briefly. An efficient, preventive anti-aging intervention may seriously contribute to successful human aging by improving the overall health status and delaying losses of brain performance.
As the pathological changes in Alzheimer dementia include degeneration of cholinergic neurones, several strategies have been used to enhance the activity of remaining cholinergic neurones, including the administration of anticholinesterases. Other drugs which have been claimed to have a beneficial effect on cognitive impairment in the aged and in various diseases include dihydroergotoxine (the most widely used in clinical practice), bufluomedil, piracetam and centrophenoxine. These drugs, and the many other compounds which have been investigated, have a wide variety of actions. However, drugs which improve the function of neuronal populations affected by pathogenic mechanisms can only be palliative. Future strategies for drug treatments will be directed towards reducing the adverse effects of the ageing process (which may include an increased susceptibility to a range of diseases) and neutralizing the pathogenic mechanisms related to genetic factors, infections, toxins, trauma, anoxia and diet. One mechanism which may operate in ageing and in some pathological changes is the production of ‘free radicals’, which can cause cellular damage. Prophylactic administration of drugs with antioxidant activity can reduce adverse effects of this process in animal models, and such treatments may also protect the brain from the effects of anoxia and other pathogenic mechanisms. Other drugs with therapeutic potential include antagonists of the excitatory effects of glutamate and aspartate.
1.1. Three methods have hitherto been applied for age pigment quantification: (a) numerically from micrographs; (b) fluorimetrically from histological sections; (c) spectrofluorimetrically from dissolved solved age pigments.2.2. The spectrofluorimetric method is at present the most commonly used technique for quantification of age pigments.3.3. By comparing the related publications since introduction of the spectrofluorimetric method, it has become apparent that few authors specify fluorimetrically significant factors, such as temperature and pH which influence the sample fluorescence during measurement.4.4. Recent developments in fluorimetrical age pigment quantification using chloroform/methanol as solvent have additionally revealed the necessity to measure and present the age pigments dissolved in both phases: the polar and non-polar solvents.
A double blind, comparative, parallel and randomized clinical trial was used for evaluation of two nootropics with anti-aging actions: Meclofenoxate (MF) and Antagonic-Stress (AS). Sixty-three old persons divided into 2 groups (average age: 68.6 and 70.8 years, respectively) with senile dementia of Alzheimer type (SDAT), of mild to moderate intensity (criteria of DSM-III-R, APA, 1987; and ICD-10, WHO, 1990) were treated with one of these nootropica. Baseline and final psychogeriatric symptomatology after three months of treatments were multiply assessed: psychogeriatric by Sandoz Clinical Assessment-Geriatric scale, Self-Assessment Scale-Geriatric and their subscales; psychometric by Wechsler Memory Scale and Wechsler Adult Intelligence Scale. Prolonged treatments with MF and AS significantly decreased the psychogeriatric scores in scales and subscales, improved the cognitive performance (attention, concentration, memory, performance IQ, full IQ) and diminshed the deterioration index (ANOVA). Therapeutical effects of AS (a neurometabolic complex containing MF) were significantly superior against MF alone (ANCOVA). MF and AS actions are discussed in connection with the brain cholinergic system, lipid peroxidation and free radical scavengers, deceleration of the aging rate, brain and erythrocyte lipofuscinolysis, multiple anti-oxidant formula, multivitamin and multimineral supplementation and with the superiority of multitherapy versus monotherapy in senile dementia and for improving the IQ and the maladaptative behavior.
Although ageing is a natural wear and tear phenomenon, it can at least be postponed or prevented by certain approaches. Some chemicals that are present in the diet or in dietary supplements have been documented to have anti-ageing effects. Recently, a number of synthetic drugs used for other therapeutic indications have been shown to have anti-ageing potential.
Oxidative stress has been implicated in the etiology of Parkinson's disease (PD). The important biochemical features of PD, being profound deficit in dopamine (DA) content, reduced glutathione (GSH), and enhanced lipid peroxidation (LPO) in dopaminergic (DA-ergic) neurons resulting in oxidative stress, mitochondrial dysfunction and apoptosis. Rotenone-induced neurotoxicity is a well acknowledged preclinical model for studying PD in rodents as it produces selective DA-ergic neuronal degeneration. In our previous study, we have shown that chronic administration of rotenone to rats is able to produce motor dysfunction, which increases progressively with rotenone treatment and centrophenoxine (CPH) co-treatment is able to attenuate these motor defects. The present study was carried out to evaluate the antioxidant potential of CPH against rotenone-induced oxidative stress. Chronic administration of rotenone to SD rats resulted in marked oxidative damage in the midbrain region compared to other regions of the brain and CPH co-treatment successfully attenuated most of these changes. CPH significantly attenuated rotenone-induced depletion in DA, GSH and increase in LPO levels. In addition, the drug prevented the increase in nitric oxide (NO) and citrulline levels and also enhanced the activity of catalase and superoxide dismutase (SOD). Histological analysis carried out using hematoxylin and eosin staining has indicated severe damage to mid brain in comparison to cortex and cerebellum and this damage is attenuated by CPH co-treatment. Our results strongly indicate the possible therapeutic potential of centrophenoxine as an antioxidant in Parkinson's disease and other movement disorders where oxidative stress is a key player in the disease process.
Previous studies in our and other laboratories indicated that there is a reduction in the neuronal lipofuscin in old rodents after several weeks of treatment with centrophenoxine. The present study investigates whether this chemical can prevent pigment formation if given early in life before the onset of pigmentogenesis. The study shows that the drug did not stop lipofuscin formation in 1 month old mice. But there was a consistent decrease in the pigment in the neurons of cerebral cortex and hippocampus of the treated animals compared to the age-matched controls. The degree of reduction was largely dependent on the duration of the treatment and a significant diminution was noted after treatment for five months or more.
Literature on aging populations of rodents supports the intuitive view that significant functional variation exists among like-aged, elderly individuals: chronological age as a solitary measure is a poor indicator of biological age. In this report, we review a variety of studies which classify aged rodents based on genetic and/or behavioral similarities, in addition to chronological age, and have provided valuable neurobiological and physiological information on age-related changes which accompany functional impairments, or the lack of them. Beyond their descriptive value for gerontological research, these findings suggest ways in which biological aging can be manipulated to promote good function in aged individuals.
The concentrations of the three major cellular forms of dolichol (free, esterified and phosphorylated) were determined in murine liver, kidney and heart. The tissue levels of these forms of dolichol were studied in detail as a function of age. Changes in the activities of dolichyl phosphate phosphatase and dolichol kinase were also determined. In liver, the concentration of unesterified dolichol, fatty acyl dolichol and dolichyl phosphate increased markedly over a period of 6 to 25 months (four-fold, 5.5-fold and nine-fold, respectively). In kidney only, free dolichol and phosphorylated dolichol increased (approximately four-fold in each case). However, this tissue consistently showed the highest concentrations of all forms of dolichol as compared to liver and heart. In heart, both free and esterified dolichol concentrations increased (approximately 3.25-fold in each case); dolichyl phosphate levels were not determined in this tissue. In all tissues studied, the activity of the dolichyl phosphate phosphatase enzyme was considerably higher than that of the dolichol kinase enzyme. In liver, there was no evidence to suggest that either enzyme was critical in determining the relative concentrations of dolichol and dolichyl phosphate. Evidence for such a role for the kinase in the kidney was stronger. Treatment of aging mice with meclofenoxate, a drug that is reported to cause dissolution of lipofuscin, failed to prevent accumulation of dolichol and dolichyl phosphate with age. These observations suggest that not all accumulated dolichol is associated with lipofuscin. Meclofenoxate treatment had no consistent effect on the activities of the enzymes studied.
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.
Lipopigment, identifiable in the fluorescence microscope, is thought to be cellular debris partly derived from free-radical-induced peroxidation of cellular constituents. The volume of neuronal lipopigment has been positively correlated with advancing age, Alzheimer dementia, and the neuronal ceroidoses, while various changes in neuronal lipopigment have been reported in association with the chronic administration of dihydroergotoxine, ethanol, phenytoin, centrophenoxine, and chlorpromazine. An increase in the volume of neuronal lipopigment may indicate increased functional activity of the cell, impaired removal of pigment or anoxia. Chronic administration of agents which can be correlated with decreased neuronal lipopigment in animal models might protect neuronal function against any adverse effects associated with (but not necessarily resulting from) lipopigment accumulation in normal ageing, anoxia, or certain degenerative diseases. Long-term studies of the prophylactic use of such agents, or of drugs which neutralise free radicals, may be indicated. Other clinical applications of such drugs may include protection against the effects of free radicals formed during periods of oxygen deprivation.
In this article the effects of neurotransmitter systems or specific drugs on cognitive functions of aged animals and humans are reviewed. While there have been used many different pharmacologic and behavioral approaches to treat and test cognitive deficits in aged animals and geriatric or demented patients, there is still the question of the validity of animal models of human disorders. Attempts are made to show that there are parallels and similarities. These similarities suggest that the neurological and neurochemical changes observed may play common roles in similar behavioral deficits observed in aged animals and humans.
Lipofuscin is defined as being a yellowish brown, lipid-rich, heterogeneous, cytoplasmic granular pigment emitting an intense yellow autofluorescence when excited with ultraviolet light, which accumulates in various tissues of animals during their aging. It is believed that the pigments are derived from the reaction of some of reactive secondary products including malonaldehyde, formed during membranous lipid peroxidation, with amino groups of phospholipids and proteins, etc., and that these formations are accompanied by alteration of the membrane structure and inactivation of the enzymes. The fluorescence measurement of the pigments is widely used as a parameter of lipid peroxidation in vivo as well as in vitro. However, their origin, chemical structure, biological significance or fate has not as yet been fully elucidated. This article introduces and discusses the recent studies on these problems.
The effect of a single, 40 micrograms, intracerebroventricular injection of colchicine on the distribution of neuronal lysosomes and lipofuscin granules in aged mice was studied. At the light microscope level we observed that colchicine induced a redistribution of dipeptidyl aminopeptidase II (Dpp II), a lysosomal and lipofuscin granule marker enzyme, from the cell bodies of neurons to the dendrites; cell bodies became depleted of Dpp II while dendrites became enriched with this enzyme. Quantitation of this phenomenon at the electron microscope level demonstrated that colchicine induced a rapid and significant decrease in the density of lysosomes and lipofuscin granules from the somata of neurons whereas in dendrites we observed a significant increase in the density of these organelles.
Intraneuronal lipopigment accumulation is associated with ageing and certain diseases, and there are many claims that this can be influenced by drugs, particularly meclofenoxate (centrophenoxine). The various unsubstantiated or conflicting reports of the effects of this drug in animal studies indicate the need for methods for the demonstration of lipopigment accumulation in adequately defined, easily-identified, and relatively homogeneous neuronal populations; this study has validated two such methods by demonstrating significant differences between groups of rats at different ages in respect of measured lipopigment autofluorescence intensity from the most heavily pigmented regions of a subpopulation of Purkinje cells, and of the area overlying intraneuronal lipopigment in a region of the hippocampus. These methods were then used to investigate the effects of daily (5 days per week) intraperitoneal injections of meclofenoxate or dihydroergotoxine, over a period of 12 weeks, before sacrifice at 13.5 months. No significant effects of meclofenoxate were detected, but dihydroergotoxine administration was associated with a significant increase in mean area overlying intraneuronal lipopigment in the CA3a region of the hippocampus. The results do not confirm that meclofenoxate can induce a reduction in intraneuronal lipopigment, but suggest that chronic dihydroergotoxine administration was associated with an increase in intraneuronal volume of lipopigment in the cell bodies of CA3a hippocampal neurones.
The ADP-Fe(II)-H2O2 system generates OH free radicals which can be trapped by 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) thus yielding a measurable signal by electron spin resonance spectroscopy. The amount of DMPO-OH spin adduct formed under certain conditions decreased considerably, if dimethylaminoethanol (DMAE), p-chlorophenoxyacetic acid (PCPA) or centrophenoxine (CPH) were present in comparable concentrations to that of DMPO. It has been demonstrated that such an effect cannot be attributed to any interference of the tested compounds with the Fe(II) and its oxidability by H2O2. The reaction of DMAE with OH free radicals was demonstrated also by using other spin traps. These spin traps reacted with OH free radicals either not at all (phenyl-tert-butylnitrone, PBN) or only to a slight extent (alfa-pyridyl-l-oxide-N-tert-butylnitrone, 4-POBN). DMAE was also a competitive OH free radical scavenger with proline and hydroxyproline, both of which have recently been shown to react with OH free radicals to form nitroxyl free radicals. On the basis of the experimental results, the OH free radical scavenger property of DMAE can be regarded as firmly established. This result supports the molecular mechanism proposed for the explanation of the anti-aging effects of CPH in terms of the membrane hypothesis of aging.
The rates of total and polyA+ RNA (mRNA) synthesis were measured by radioisotope technique in the brain cortex of female CFY rats. There was practically no significant difference between the young (1.5 months) and adult (13 months) rats; however, the old group (26 months) displayed a considerable decrease of the rates of synthesis of both classes of RNA studied. Centrophenoxine treatment (100 mg per kg body weight per day, for 2 months) reversed this tendency, and increased significantly the synthesis rates of old rats almost to the adult level. The results are interpreted in terms of the membrane hypothesis of aging, attributing a free-radical scavenger function of the dimethylamino-ethanol incorporated into the nerve cell membrane from the centrophenoxine.
The effects of centrophenoxine on the retinal pigment epithelium (RPE) of 17 month old female mice have been studied. Animals were injected subcutaneously for 3 months (60 injections) with the drug (0.1 mg/g of body wt) daily in 0.1 M phosphate buffered saline at pH 7.0. The morphological changes in the pigment layers of the retina of both eyes were studied by light and electron microscopy and the lipofuscin pigment was demonstrated by its autofluorescence and ultrastructural characteristics. There was a significant reduction of the lipofuscin pigment in the treated animals, but the melanin pigment remained unchanged. The lipofuscin granules also appeared less osmiophilic and showed a greater preponderance of membranes and vacuoles. Although the precise mechanism of action of the drug is not clear, an increased protective function of the pigment epithelium by the drug has been suggested.
Young, adult and old female Wistar rats (3, 18 and 28 months of age, respectively), were studied using electron-microscopic stereology. Synaptic parameters of the cerebellar glomerulus were calculated and compared with similar data obtained from old Wistar rats of the same breed treated with centrophenoxine (CPH; HelferginR, Promonta, Hamburg) in the form of intraperitoneal injections (100 mg/kg body weight) for 40 days. This treatment resulted in a sort of "rejuvenation" of synaptic structures. Namely, the surface density and the total length of synaptic contact zones were markedly reduced in the untreated old group, but in the treated animals these parameters returned to the values found in the young and adult animals. At the same time the numerical density of synapses remained unaltered in the treated group, while the average synaptic length displayed some further increase. The results are interpreted in terms of the age-dependent decrease in reactive synaptogenesis, suggesting that CPH stimulates the metabolism of the nervous elements persisting in old brain. The possible mechanism of CPH effect is also discussed.
The cerebral cortex and liver tissues of young, adult and old (1–4, 15 and 25–30 months of age, respectively) CFY rats of both sexes were investigated. Water-soluble (WSP) and water-insoluble (WIP) protein fractions were separated after homogenization of 10 mg tissue per 1 ml of distilled water, by centrifugation at 500 g. The WSP and WIP fractions were analyzed by electron microscopy and their protein contents were determined by a modified Folin phenol reaction. Electron microscopy revealed a complete destruction of the cell organelles due to the strong ostomic shock. Total protein contents of both tissues increased significantly in the old rats and also the WIP contents were much higher in aged animals than in the young and adult ones. Mild heat treatment (64°C, 10 min) or 3 M urea were able to resolubilize a considerable portion of the WIP in the young animals, however, in adults and olds their effect was strongly reduced. Protease inhibitors displayed only a very limited influences on the WIP. The conclusion was reached that a considerable portion of WIP of the old animals may be covalently cross-linked. Certain methodical problems of protein determination are also discussed. In vivo application of centrophenoxine (100 mg/kg body weight) for 4 weeks significantly reduced the WIP content in 25-month-old females, and the total protein content also displayed a decreasing tendency. The results are discussed in terms of the membrane hypothesis of ageing.
Summary The cerebral insufficiency improvers centrophenoxine, piracetam and hydergine were tested for their effect on lipid peroxidation of rat brain homogenates, both in vitro and in vivo. There was no effect either in vitro or after chronic 8 week dosing of animals.
The cerebellar glomerulus was studied by electron microscopic morphometry in female Wistar rats. Age-dependent alterations have been revealed from 3 to 28 mth of age, and the effect of centrophenoxine (CPH) was analyzed in two different patterns of administration. First, 27-mth-old rats were treated daily for 6 wk (acute treatment), and second, 18-mth-old rats were treated 3 times per week for 5 months (chronic treatment). The dose was 100 mg CPH/kg body weight, injected intraperitoneally. The surface density (SV), the numerical density (NV) and the average length (L) of the synaptic junctions were calculated from data obtained on ethanol-phosphotungstic acid stained ultrathin sections. An age-dependent reduction of SV and NV of the synaptic contact zones was found, and the L increased in the oldest animals. CPH-treatment resulted in a marked increase of SV in both types of application, whereas the other two parameters behaved differently in the two groups. The chronic treatment resulted in a significant slowing down of the decrease of NV, whereas L remained invariate. On the contrary, the acute treatment increased L but did not alter significantly NV. The results and the differences between the treatment types are discussed in terms of synaptic plasticity and are interpreted as different manifestations of the same reactive synaptogenetic process.
The amount of intraneuronal lipofuscin in the parietal cortex and in the inferior olivary nucleus was measured in post-mortem tissue affected by Alzheimer dementia and in specimens from non-demented individuals. The results indicate that there is a liner relationship between the accumulation of cell body lipofuscin and advancing age, both in neuronal populations of the non-demented groups, and in the olivary neurones of the demented group. However, in the demented group, the lipofuscin fluorescence intensity of the parietal neurons was not significantly correlated with age. The estimated amount of lipofuscin in the olivary neurons in the demented group was significantly higher than in the non-demented group, when age has been taken into account (P less than 0.01). The possible pathogenic role of lipofuscin accumulation in ageing and Alzheimer dementia is discussed.
Water-insoluble protein fractions increase in the brain cortical tissue and liver of rats during aging in both sexes. This suggests a possible increase in the cross-linking of proteins which may be due to the formation of, for example, hydroxyl free radicals during several metabolic processes. In vivo application of centrophenoxine causes a reversal of this phenomenon in old rats. In vitro experiments show that the generation of hydroxyl free radicals by chemical systems like homolysis of H2O2 by redox coupling with Fe2+ leads to Fe3+ conversion, results in the cross-linking of bovine serum albumin and the mixed proteins of liver or brain homogenates of young rats. The cross-linked proteins have a very much increased molecular weight, they become mostly insoluble even in 6 M urea. Dimethylaminoethanol, the effective part of the centrophenoxine molecules, is able to diminish the extent of cross-linking, acting most probably as a free-radical scavenger. The results are discussed in terms of the "membrane hypothesis of aging". A molecular basis is proposed for the anti-aging effect of centrophenoxine.
Vitamin E is known to play a protective role for cell membranes against free-radical attacks. Vitamin E deficiency causes a rapid macroscopic ageing of rats. On the other hand, during normal ageing, cell membranes undergo functional alterations resulting in an increased intracellular potassium concentration in brain and liver cells. Therefore, is was of interest to study whether vitamin E deficiency produces similar alterations in young rats. Female Wistar rats were fed with a vitamin E deficient diet from 1 month of age for 10 months. The parietal brain cortex and the liver were analyzed by means of a quantitative energy dispersive X-ray microanalytic method using a JEOL JSM-35C-EDAX-711-NOVA-3 system. Monovalent electrolyte contents as well as the water content of the cells were determined in 5 treated and 5 control animals. Water content was measured by analyzing the potassium content in aqueous, frozen state, and again in the dry mass of the cells. On the basis of these data, a computer program calculated the water proportions. Average values for 200 or more cells of each organ per group revealed a significant increase in the intracellular potassium content of the brain cells, whereas the sodium and chloride contents increased to a much lower extent. There was a 2.6% loss of intracellular water in the brain cells in the vitamin E deficient group. The liver monovalent ions and water content remained unchanged. The results obtained are discussed in terms of the membrane hypothesis of ageing.
A complex antiaging formula--Antagonic-Stress--was investigated vs. placebo (PL), meclofenoxate (MF)--neurometabolic nootropic and vs. nicergoline (NE)--cerebral vasodilator by comparative multiple trials (double-blind, randomized, and parallel) in gerontopsychiatry (DSM-III-R, 1987 and ICD-10, 1992 criteria). AS vs. PL studies in organic mental disorders--amnestic, depressive, anxiety, associated with axis III physical disorders or conditions, and in multiinfarct dementia were followed by AS vs. MF or NE investigations in senile dementia of Alzheimer's type. A total of 343 old people, distributed in 4 PL groups, 1 MF group, 1 NE group, and 5 AS groups were studied. Multiple investigations, before and after three-month treatments were made: psychometric evaluation by Sandoz Clinical Assessment-Geriatric, Self-Assessment Scale-Geriatric and their 5 subscales; psychopathological rating by Hamilton Depression and Anxiety Scales; as well as psychometric testing by digit symbol of WAIS, Wechsler Memory Scale and Wechsler Adult Intelligence Scale (WAIS). Except PL, prolonged and large dose treatments with these cerebral activators (MF, NE and especially AS) reduced the psychogeriatric-psychopathological scores and the deterioration index, and improved cognitive performance. The therapeutical effectiveness of AS multiple formula in gerontopsychiatry and its superiority vs. monotherapy (MF or NE) are discussed in connection with its complex neurometabolic and synergetic composition, multiple antioxidative combinations, free radical scavengers, lipofuscinolytic agents, the antiischemic action of antioxidants, multivitamin and multimineral supplementation, and with the better efficacy of multitherapy vs. monotherapy in geriatrics.
alpha-Lipoic acid (alpha-LA) improved longer-term memory of aged female NMRI mice in the habituation in the open field test at a dose of 100 mg/kg body weight for 15 days. In a separate experiment, no such effect could be found for young mice. alpha-LA alleviated age-related NMDA receptor deficits (Bmax) without changing muscarinic, benzodiazepine, and alpha 2-adrenergic receptor deficits in aged mice. The carbachol-stimulated accumulation of inositol monophosphates was not changed by the treatment with alpha-LA. These results give tentative support to the hypothesis that alpha-LA improves memory in aged mice, probably by a partial compensation of NMDA receptor deficits. Possible modes of action of alpha-LA based on its free radical scavenger properties are discussed in relation to the membrane hypothesis of aging.
Changes in lipid peroxidation, lipofuscin concentration, and multiple unit activity (MUA recorded in conscious animals) in the CA3 region were studied in the hippocampus of male Wistar rats aged 4, 8, 16, and 24 months. The lipid peroxidation and lipofuscin concentration were increased with age. The MUA, however, declined with age. Correlational analyses were performed for the four age groups to determine the relationship between the age-associated decline in MUA with the age-related alterations in lipid peroxidation and lipofuscin concentrations. The age-related increase in lipid peroxidation correlated positively with the age-associated increase in lipofuscin concentration. The age-related increases in lipid peroxidation and lipofuscin concentration correlated negatively with the changes in MUA. Since lipid peroxidation may affect neuronal electrophysiology, our data suggested that age-related increase in lipid peroxidation may contribute to an age-associated decline in neuronal electrical activity. Centrophenoxine effects were studied on the three above-mentioned age-associated changes in the hippocampus. The drug had no effect on all three parameters in 4- and 8-month-old rats. In 16- and 24-month-old rats, however, the drug significantly increased the MUA but concomitantly decreased lipofuscin concentration and lipid peroxidation. Correlational analyses of the data on MUA, lipid peroxidation and lipofuscin concentration from the centrophenoxine-treated animals showed that the drug-induced diminution in both lipofuscin and lipid peroxidation was significantly correlated with the drug-induced increase in MUA. The differential effect of the drug in younger (4-8 months) and older (16-24 months) animals indicated that the stimulation of MUA was clearly associated with concomitant decrease in lipid peroxidation and lipofuscin concentration.
In both stress and aging, an etiopathogenic analysis demonstrates in the brain some common mechanisms and reciprocal accelerating relationships: hypoanabolism (decrease of RNA and protein synthesis), coupled with hypercatabolism (increase of oxidative stress-lipid peroxidation, with waste product accumulation: lipofuscinage pigment and water-insoluble proteins). For therapeutical intervention in these antihomeostatic processes, we successively (1972-1992) developed a neurometabolic antioxidative therapy--finally represented by a specific antistress and antiaging synergistic formula, Antagonic-Stress. Its homeostatic actions have been demonstrated in the stressed aging brains by reestablishing the anabolism/catabolism balance: anabolic regeneration by increasing total RNA, total proteins and water-soluble proteins, coupled with catabolic regulation by accelerated lipofuscinolysis and age pigment elimination (neuronal-->glial-->capillary route) and decreasing of water-insoluble proteins. Specific, synergistic, and superior actions of Antagonic-Stress multiple formula vs. antistress and antiaging monotherapy have been demonstrated by preclinical and clinical studies, confirming our results.
As was shown in a recent review by this author (Ann. N.Y. Acad. Sci., 928: 187-199, 2001), oxyradicals cannot be considered only as harmful by-products of the oxidative metabolism, but living cells and organisms implicitly require their production. This idea is supported by numerous facts and arguments, the most important of which is that the complete inhibition of the oxyradical production by KCN (or by any block of respiration) kills the living organisms long before the energy reserves would be exhausted. This new theoretical approach not only helps our understanding of the normal functions of the living organisms, such as the basic memory mechanisms in the brain cells, but also helps in identifying the site-specific, radical-induced damaging mechanisms that represent the undesirable side effects of oxygen free radicals. First of all, these effects make the cell plasma membrane vulnerable and cause a series of intracellular functional disorders, as described by the membrane hypothesis of aging (MHA). The logical way for any antiaging intervention therefore should be to increase the available number of loosely bound electrons inside the plasma membrane that are easily accessible for OH(*) free radical scavenging. The present review summarizes the available knowledge regarding the theory of the use of membrane-related antiaging pharmaca, like centrophenoxine (CPH), tested in both animal experiments and human clinical trials. A modified, developed version of CPH coded as BCE-001 is also reported.
The environmental agent aluminium has been intensively investigated in the initiation and progression of various neurological disorders and the role of oxidative stress in these disorders is a widely discussed phenomenon. In this light, the present study is focused on the role of aluminium in mediating oxidative stress, which may help in better understanding its role in neuronal degeneration. Further, we have exploited a known anti-aging drug centrophenoxine to explore its potential in the conditions of metal induced oxidative damage. Aluminium was administered orally at a dose level of 100 mg/kg b.wt./day for a period of 6 weeks followed by a post treatment of centrophenoxine at a dose level of 100 mg/kg b.wt./day for another 6 weeks. Following aluminium exposure, a significant increase in lipid peroxidation levels (estimated by MDA) were observed which was accompanied by a decrease in reduced glutathione content in both cerebrum and cerebellum of rat brain. Post treatment of centrophenoxine significantly reduced the lipid peroxidation levels and also increased the reduced glutathione content in both the regions. Histologically observed marked deteriorations in the organization of various cellular layers in both cerebrum and cerebellum were observed after aluminium administration. Centrophenoxine treated animals showed an appreciable improvement in the histoarchitecture of the cellular layers. Our results indicate that centrophenoxine has an antioxidant potential and should be examined further in aluminium toxic conditions.
Autonomic and sensory ganglion cells in the senile dog contain a deposition of PAS-positive substances which has been shown to be mucoprotein in nature.
Data are presented to show that this PAS-positive mucoprotein can be demonstrated by metachromatic staining with toluidine blue after the mucoprotein is sulfated. This procedure indicates that mucoprotein is also present in a granular form in all nerve cells in both senile and young dogs.
The evidence for this is further substantiated by the use of the aldehyde-fuchsin stain following both periodic acid oxidation and sulfation. The granular and non-granular deposition can be demonstrated by the periodic acid-aldehyde-fuchsin method due to the affinity of the aldehyde-fuchsin stain for aldehydes. It can be demonstrated following the sulfation-aldehyde-fuchsin method owing to the affinity of the stain for the sulfuric group. The evidence for this latter phenomenon has been reported by Scott and Clayton (6).
It is concluded that mucoprotein is present in a granular form in all nerve cells in both senile and young dogs but is not concentrated enough in the latter to be demonstrated by the PAS method.
An examination of the topographic distribution of lipofuscin pigment granules with the light and electron microscope revealed either smaller and randomly "dispersed" or larger and more complex "clustered" pigment configurations in the cytoplasm of neurons in the dorsal ganglia and ventral spinal cord of 24-month old male mice. Qualitative comparisons revealed no major differences in shape, size, complexity, density, orientation, and cytologic distribution of the pigment bodies in motor and sensory neurons. In general, when the pigment granules were quite numerous within the 2 types of cells, they were smaller in size ( approximately lmicro), had a dense homogeneous matrix with few bands or lamellae, and were uniformly distributed throughout the cytoplasm. In contrast, when the pigment configurations were less in number, they were usually larger in size ( approximately 3micro), had a more complex internal banded structure, and appeared more localized within the cell. Examination of the bands revealed a repeating pattern of approximately 70 A. The bands appeared to fuse, forming hexagonal arrays of linear densities intersecting at an angle of approximately 120 degrees in some regions of the pigment bodies. Structural similarities suggested that the striated membranous bands may be composed of phospholipids.
Two distinctly different groups of disorders have emerged from the conditions generically classified as amaurotic familial idiocies. One is characterized by grossly abnormal profiles for cerebral sphingolipids, for example the GM1 and GM2 gangliosides. The other group is composed of patients with normal sphingolipid profiles, but with neuronal accumulation of lipopigments of the ceroid-lipofuscin type. The sphingolipidoses have been shown by a number of investigators to meet the classic concept of Hers (1965) for lysosomal diseases. This view has been repeatedly reinforced by continuing studies which show the lack or reduction of specific hydrolases, resulting in the accumulation of biochemical compounds which cannot be degraded to metabolically utilizable substances. The concept of lysosomal diseases has led many investigators to search for a single accumulating lipid or a deficient hydrolytic enzyme unique to neuronal ceroid-lipofuscinoses. Since many of the lysosomal disorders became better understood by the elucidation of the chemical properties of a specific lipid, present in unphysiologically large quantities, the pronouncement by Donahue et al. (1966) before this group, that the “chemical analysis of cytoplasmic lipopigment granules (from the brains of patients with Batten’s disease) will contribute little towards an understanding of this disease” was decidedly unwise. Although it is correct, that autofluorescent lipopigments lack chemical specificity and result from a great variety of pathogenetic situations (Porta and Hartroft, 1969), Donahue et al. (1966) failed to recognize the distinct possibility, that certain repetitive patterns in the chemical composition and ultrastructure of these residual bodies may indicate a specific formative pathogenesis. On the strength of this argument, we developed methods to isolate lipopigments in pure preparations and the first results have proved already the soundness of this concept. As it turned out, the previously held concept of a close relationship between lipofuscin and ceroid had to be abandoned and replaced by the theses that these classes of lipopigment are distinctly different entities, albeit both represent residual bodies and both contain polymeric substances (Siakotos et al., 1970).
C1300 mouse neuroblastoma cells in culture were used to study cytoplasmic lipofuscin pigment accumulation and the ability of centrophenoxine to alter its formation. The pigment was detected by its autofluorescence, by histochemical staining for acid phosphatase, and by positive staining with the periodic acid Schiff procedure. The percentage of cells with pigment increased from 25% at 5 days of culture to 60% at 25 days of culture. Pigment formation was enhanced by treatment for 9 days with either 1.1x10-5M papaverine or 5.6x10-5M prostaglandin E1. Pigment formation in papaverine treated cells was markedly reduced by treatment for 9 days with either 1.0 or 3.4x10-4M centrophenoxine. It is concluded that neuroblastoma cells in culture provide an in vitro model with which to study lipofuscin pigment formation and its manipulation by pharmacological agents.
Contrary to the earlier light microscopic studies, electron microscopic investigation on 11 male guinea pigs revealed marked accumulation of lipofucsin pigment in the hypothalamic neurons with the passage of time. Even in 6 month guinea pigs scattered pigment bodies were noted. Two types of pigment granules have been observed in the aged animals. Dimethylaminoethyl p-chlorophenoxyacetate caused a marked diminution of the electron density and reduction in the size and number of the pigment granules. Furthermore, histochemical examination revealed a marked diminution of acid phosphatase activity in the treated group of animals as compared to normal "control". But the α-esterase and succinic dehydrogenase activity was found to be increased following drug treatment.
Mit histochemischen Methoden lassen sich in Lipopigmenten (Lipofuscin und Ceroid) hydrolytische Enzyme, und zwar saure Phosphatase und unspezifische Esterase nachweisen.Bei den Lipofuscinen erfolgt die Pigmentbildung hauptschlich in Parenchymzellen, und zwar offenbar an Stellen im Cytoplasma, welche im Stoffwechselgeschehen eine besondere Rolle spielen und von vornherein eine ausgesprochene Enzymaktivitt besitzen oder ganz selektiv zur Fermentbildung befhigt sind.Das Ceroid wird dagegen anscheinend in Phagozyten abgelagert, welche primr keine deutliche Fermentaktivitt besitzen. Offenbar ist hier die Enzymaktivitt eine Reaktion auf die Ablagerung von Fettstoffen.
Aufbau und Vorkommen des Alterspigmentes in Neuronen und Satellitenzellen der cervicalen Spinalganglien seniler Ratten wurde licht- und elektronenmikroskopisch nach 4-, 5-, 8-, 11- und 13 wöchiger Centrophenoxinbehandlung untersucht und mit dem in gleichaltrigen Kontrolltieren verglichen. Lichtmikroskopisch fand sich am PAS-gefärbten Material eine Verminderung des Alterspigmentes nach 8 wöchiger und längerer Behandlungsdauer. Elektronenmikroskopisch ließen sich bereits nach 4 wöchiger Centrophenoxinbehandlung vacuolige Veränderungen an den Pigmentkörpern der Satellitenzellen feststellen. Gleichartige Veränderungen zeigten sich nach 8 wöchiger Behandlung auch an den Pigmenten der Nervenzellen. Gleichzeitig fanden sich die Pigmentkonglomerate in den Typ A-Neuronen in einem Zustand der Disintegration. Die Veränderungen nahmen mit der Dauer der Behandlung zu und führten zu einer Verminderung der Pigmentkörper. Die möglichen Ursachen der Veränderung bzw. Verminderung der Pigmentkörper werden diskutiert und der Weg eines etwaigen Abtransportes der Alterspigmente aus den Nervenzellen erörtert.
The physical and chemical properties of the lipofuscin pigment have been studied in the neurons of C57 BL/10 mice of different ages. The results indicated the presence of two types of lipofuscin with different physical and chemical properties. The properties of these two types of lipofuscin have been discussed. It appears that the two types of lipofuscin found in young and old mice differ in their distribution, stainability, solubility, enzymatic activity, and fluorescence properties. It is suggested that these represent early and late stages in lipofuscin formation of the neurons. The pigment observed in animals deprived of Vitamin E was atypical in nature and did not conform to either of the types mentioned above.
Dimethylaminoethyl p-chlorophenoxyacetate increased the median, mean and maximum survival time from the start of drug administration of male Swiss Webster Albino mice by 29·5 per cent, 27·3 per cent (P = 0·039) and 39·7 per cent respectively. The drug treatment was associated with some loss of weight. The drug was previously reported to reduce lipofuscin pigments in the neurons of senile guinea pigs, while concomitantly reducing the activity of the lysosomal enzymes, acid phostase and simple esterase. This suggests that the drug is active with respect to three postulated mechanisms involving lysosomes in the aging process. Within minutes after dissolving in water, dimethylaminoethyl p-chlorophenoxyacetate breaks down into dimethylaminoethanol and p-chlorophenoxyacetic acid. The observed effects therefore may be those of the hydrolysis products.
Mutant mice are described which have an early developing locomotor difficulty accompanied by definite neuronal changes in the central nervous system. They develop head tremors during the second postnatal week and later action tremors while walking. Seizures occur spontaneously and can be induced by stimulation. By the third or fourth week, they lose the righting reflex. The most apparent neuropathologic sign is the progressive development of nuclear hyperchromasia, especially in the largest neurons of the spinal cord and brain stem. Purkinje cells of the cerebellum are similarly affected. Hyperchromasia occurs in single, isolated neurons scattered throughout the central nervous system, as well as in groups of cells which comprise a brain stem nucelus. Lipofuscin pigment in quantities comparable to that in neurons of 12 months old mice was found in neurons with hyperchromatic nuclei as early as five weeks of age, an observation which suggests that premature aging might be occurring in the mutant's central nervous system.
Pure lipofuscin has been isolated from normal human brain by a new technique based on flotations and density-gradient centrifugations procedures coupled with dextran sulfate precipitation of interfering subcellular components. This lipofuscin preparation has been shown to be made up of a number of species which may be various stages in the development of the residual body or secondary lysosome from the initiating lysosome body.A second autofluorescent pigment, ceroid, with a markedly different density, was isolated from two human cases of neuronal ceroid-lipofuscinosis and a canine brain affected with a similar disorder. This pigment was isolated by an enzymatic procedure followed by density-gradient centrifugation. Three morphologically different forms of ceroid were found for each of the three individuals affected with neuronal ceroid-lipofuscinosis.
Dimethylaminoethyl p-chlorophenoxy acetate (80 mg/kg body weight) was administered (i. m.) to guinea pigs for 30 to 56 days. Electron microscopic examination of the hippocampus, mid-brain reticular formation and the area postrema revealed marked diminution in the electron density of the pigment granules and vacuolization. This type of lipofuscin was detected in some phagocytic cells and in the capillary endothelium. Conspicuous vacuolization of the capillary wall was discernible. These changes were not observed in the control group of animals.
CLINICAL and psychological studies indicate that a decline in the functions of the nervous system occurs in old age. Although it is difficult to evaluate the physiopathological changes in the neurones in senescence, the cells nevertheless show changes which they have acquired during functional activities in the past. Neurones are particularly suitable for the study of the effects of time on living cells as these are not generally replaced during life.
Thirty eight drugs, added to the nutrient medium at one or more concentrations, were studied for their effect on life span in Drosophila melanogaster. Most drugs were selected for their known or suspected capacity to stabilize cellular membranes—particularly lysosomal membranes—against breakdown in vitro or in vivo. Median life span, maximum life span, and in selected cases mean life span, its standard deviation and the statistical significance of differences over controls were determined.Significant extensions of mean life span in both male and female flies were produced, sometimes over a range of concentrations, by cortisone (18 to 43 per cent), hydrocortisone (21 to 39 per cent), triamcinolone (18 to 19 per cent), aspirin (13 to 40 per cent), salicylamide (12 to 17 per cent), a 3:1 mixture of salicyl-salicylic acid plus aspirin (24 to 38 per cent), acetaminophen (5 to 9 per cent) and meclofenoxate (7 to 39 per cent). Appreciable extensions of maximum life span were also noted for these drugs. Diphenhydramine, chlorpheniramine, doxylamine, tripelennamine and colchicine also produced moderate extensions of median and maximum life span. Other drugs produced lesser, inconsistent, or insignificant results or shortened life span at one or more dosage levels.RésuméL'auteur étudie l'influence sur la longévité de Drosophila melanogaster de 38 produits ajoutés en doses variables au régime alimentaire. La plupart de ces produits avaient été choisis à cause de leur aptitude, connue ou présumée, à stabiliser les membranes cellulaires et plus particulièrement les membranes lysosomales contre la dégradation in vitro ou in vivo. L'auteur a déterminé la longévité médiane, la longévite maximale et, dans certains cas choisis, la longévité moyenne, son écart standard et la signification statistique des différences par rapport aux individus témoins.Des augmentations significatives de la longévité moyenne des mouches, mâles et femelles, ont été obtenues, parfois pour une gamme de dosages, avec la cortisone (de 18 à 43 p. 100), l'hydrocortisone (de 21 à 39 p. 100), la triamcinolone (de 18 à 19 p. 100), l'aspirine (de 13 à 40 p. 100), le salicylamide (de 12 à 17, p. 100), un mélange à 3:1 d'acide salicyl-salicylique et d'aspirine (de 24 à 38 p. 100), l'acétaminophène (de 5 à 9 p. 100) et le méclophénoxate (de 7 à 39 p. 100). Ces produits ont aussi augmenté notablement la longévité maximale. La diphénhydramine, la chlorophéniramine, la doxylamine, la tripelennamine et la colchicine aussi ont augmenté modérément les longévités médiane et maximale. D'autres produits ont donné des résultats moins réguliers ou insignifiants ou ont abrégé la longévité à un ou plusieurs niveaux de dosage.ZusammenfassungDer Effekt von 38 Drogen auf die Lebensdauer von Drosophila melanogaster wurde untersucht, indem diese Stoffe dem Nährmedium in einer oder mehreren Konzentrationen zugesetzt wurden. Die meisten der Drogen wurden wegen der bekannten oder vermuteten Fähigkeit zur Stabilisierung von Zellmembranen, insbesondere lysosomalen Membranen, hinsichtlich dem in-vitro-oder in-vivo-Abbau ausgewählt. Die Durchschnittslebenszeit, die maximale Lebensdauer und in ausgewählten Fällen die mittlere Lebensdauer mit Standardabweichung und der statistischen Signifikanz von Abweichungen gegenüber Kontrollwerten wurden bestimmt.Signifikante Verlängerung der mittleren Lebensdauer bei männlichen und weiblichen Fliegen wurde erzielt, manchmal über einen Konzentrationsbereich, mit Cortison (18 bis 43 Prozent), Hydrocortison (21 bis 39 Prozent), Triamcinolon (18 bis 19 Prozent), Aspirin (13 bis 40 Prozent), Salicylamid (12 bis 17 Prozent), 3:1 Mischung von Salicyl-Salicylsäure plus Aspirin (24 bis 38 Prozent), Acetaminophen (5 bis 9 Prozent) und Meclofenoxate (7 bis 39 Prozent). Für diese Drogen wurde auch eine erhebliche Verlängerung der maximalen Lebensdauer beobachtet. Diphenhydramin, Chlorpheniramin, Doxylamin, Tripelennamin und Colchizin verursachten auch eine mäßige Verlängerung der mittleren und maximalen Lebensdauer. Andere Drogen erzeugten geringere, nichteinheitliche oder nichtsignifikante Veränderungen oder verkürzten die Lebensdauer bei einer oder mehreren Konzentrationen.РезюмеИсследовалось влияние тридцати босьми лекарственных средств, прибавленных в различных концентрацичх к питательной среде, на длительность жнзни Drosophila melanogaster Большинство медикаментов выбиралось по их известной или подозревае мой способности стабилизировать клеточные мембраны—в особенности лизосомных оболочск—против распада in vitro или in vivo. Определялись—медиана продолжнтельности жизни, максимальная продолжительности жизни, а в избранных случаях—средняя продолжительность жизни, а также станлартное отклонение и статистическая значимость разницы по сравнению сонтролями.зчачимыс удлкнения средней продолжительности жизни у мух, как самцов так и са⇐ок обнаруживались, иногда при различных концентрациях, под влиянисм кортизоиа (18–43%), гидрокртизона (21–39%), триамцинолона (18–19%), аспирина (13–40%), цслициламида (12–17%), 3:1 смеси салицила-салициловой кислоты с аспирином (24–38%), ацетаминофена (5–9%) и меклофеноксата (7–39%). Эти препараты также значительно продлили максияальную продолжительности жизни. Умеренное продление медианной и максима←ьной длительности жизни были тоже получены сдифенгндрамидом, хлорфениламином, доксиламином, трипеленнамином и колшицином. Другие препараты вызвали меньшие противоречивые или незначительные результаты или зе укоротили длительность зизни при той или другой дозировке.
The studies reviewed here reflect a marked difference of opinion among investigators on some fundamental aspects of lipo-fuscin research, such as its correlation to ageing, different type(s) and origin(s) of pigment granules, role of trauma and diseases, and the effect of drugs. An attempt has been made to present a unified concept of origin of lipofuscin and to point out the lacunae in our knowledge with a view to stimulate further research.Copyright © 1972 S. Karger AG, Basel
Lipofuscin formation in the neurons of spinal ganglia of one month old male Wistar rats, maintained in culture for a period of 4 weeks, has been investigated. The dorsal root ganglia were explanted on collagen coated coverslips and carried in Maximow double coverslip assemblies. The cultured ganglia were studied in sections after OsO4 fixation and Epon embedding. The age pigment is found as early as one month in spinal ganglia neurons. In culture an accelerated accumulation of lipofuscin occurs and after four weeks of maintenance of the spinal ganglia, the neurons at different rates degenerate. The pigment bodies, some vacuolated others not, are often seen to be bound by a double limiting membrane and are found in close topographical and morphological association to mitochondria. The pigment from the ganglion cells can be removed by cytoplasmic protrusions which accounts also for the presence of pigment bodies in the satellite cells.
This chapter focuses on the behavioral and neuropathological manifestations of nutritionally induced central nervous system “aging” in the rat. The long-term objective of the present study is to determine any relationship between the accumulation of central nervous system (CNS) lipofuscin and the deterioration of brain functions. In the present experiment accumulation of brain lipofuscin and the behavior related to memory and learning functions were studied in rats chronically fed on a vitamin E deficient diet. Accelerated “aging” with respect to brain accumulation of lipofuscin (intracellular granules exhibiting yellow primary fluorescence) was induced in male rats (Sprague–Dawley) by feeding them an artificial diet deficient in vitamin E. Acquisition and retention of conditional-avoidance response, performance of delayed-alternate responding and retention of one-trial learning of aversive experience were impaired. During testing, the animals were placed on normal laboratory chow; therefore, the senility-like deficits of learning and memory were because of irreversible neuropathology (brain atrophy and lipofuscin accumulation) caused by aging and vitamin E deficient diet.
Structural changes in neurons in the cervical anterior horn of ageing mice have been studied. The main age-related neurocytological change observed during this study is progressive accumulation of lipofuscin. The pigment was present in some nerve cells as early as six weeks after birth. Various types of membrane-bound granules encountered in the anterior horn cells were characterized on the basis of their shape, size and fine structure into primary lysosome-like (L1) granules (dense bodies), autophagic vacuole-like (L2) granules and mature (L3) pigment granules of complex substructure and irregular configuration.L1,L3 andL3 types of granules appear to represent respectively early, intermediate and mature stages in a developmental continuum of lipofuscin pigment granules. Transitional stages suggest that matureL3 pigment granules evolve by gradual alteration of lysosome-likeL1 andL2 granules. A probable sequence of morphologic events accompanying the transformation of lysosome-like granules into mature lipofuscin pigment granules is suggested.
The localization of the lipofuscin pigment in the twelve brain areas of the pig was included in the present investigation. It was observed that pigment was present in the adult pig brain and increased with advancing age. The study was conducted on thirty pigs ranging between the ages of 2 days and 10 yr. The pigment was identified with histologic stains and autoflorescence techniques. The time of appearance of pigment varied in different brain areas, but these differences progressively diminished with advancing age. The presence of pigment formed the most consistent neurocytological age associated change in aging nerve cells.
The pathologic findings in two patients with progeria are described. The two patients had several features in common: the history of congestive heart failure preceding death, the pathologic findings of interstitial and focal myocardial fibrosis and necrosis, and intracellular accumulation of lipofuscin pigment in many organs. One patient also had chronic glomerulonephritis. A common belief has been that the cardiac lesion in progeria is typical myocardial infarction. However, even in the absence of significant coronary artery disease, focal myocardial fibrosis has been noted in previous cases as well as in the patients in this report, and, therefore, it is clear that both types of lesions occur in progeria. Finally, lipofuscin accumulation has not been noted previously in progeria. This finding lends support to the concept of progeria as a syndrome of premature aging.
In this investigation Dr. Jayne demonstrates that an acid-fast, insoluble pigment, which accumulates progressively with age,
is first formed at the poles of the cardiac muscle nuclei during the second decade of life. This paper was presented in part
at the meeting of the association of anatomists in philadelphia, april 13-15, 1949.
On the use of centrophenoxine in hospitalized and ambulant patients
- Houillon G
Versuche zur medikamentosen Beeinflussung altersbedingter Veranderungen
- Chemnitius
Therapeutic trial of centrophenoxine in manifestation and accidents of old age
- Fichez LF
Derivatives of acids acting as growth control agents in plants. Pharmacological properties of dimethylaminoethyl ester of parachlorophenoxy acetic acid
- Thuillier G
The first results of parachlorophenoxyacetate of dimethylaminoethyl (ANP235) in psychiatry
- Delay J
Histochemical studies on the lipofuscin pigments in human heart
- Hamprel H
Weiter Untersuchungen uber sichtabare Fluorezenz beim Menschem
- Bomer S
Organization of the cerebral cortex
- Brody
The occurrence and duration of senile pigment experimentally induced in the nerve cells of the young rat
- Sulkin NM
Lipofuscin pigment formation in neuroblastoma cells in culture
- Nandy