Free amino acids in motor cortex of amyotrophic lateral sclerosis.

Free amino acids were estimated quantitatively in the motor cortex from 3 patients with amyotrophic lateral sclerosis (ALS) and 11 control subjects. Among 7 amino acids which showed statistically significant changes, taurine was the only one which was increased constantly and most markedly in the motor cortex of all the 3 ALS cases. It was suggested that the metabolism of sulfur amino acids might be affected in comparatively early stages of ALS.     

Cellular therapy to target neuroinflammation in amyotrophic lateral sclerosis

Neurodegenerative disorders are characterized by the selective vulnerability and progressive loss of discrete neuronal populations. Non-neuronal cells appear to significantly contribute to neuronal loss in diseases such as amyotrophic lateral sclerosis (ALS), Parkinson, and Alzheimer’s disease. In ALS, there is deterioration of motor neurons in the cortex, brainstem, and spinal cord, which control voluntary muscle groups. This results in muscle wasting, paralysis, and death. Neuroinflammation, characterized by the appearance of reactive astrocytes and microglia as well as macrophage and T-lymphocyte infiltration, appears to be highly involved in the disease pathogenesis, highlighting the involvement of non-neuronal cells in neurodegeneration. There appears to be cross-talk between motor neurons, astrocytes, and immune cells, including microglia and T-lymphocytes, which are subsequently activated. Currently, effective therapies for ALS are lacking; however, the non-cell autonomous nature of ALS may indicate potential therapeutic targets. Here, we review the mechanisms of action of astrocytes, microglia, and T-lymphocytes in the nervous system in health and during the pathogenesis of ALS. We also evaluate the therapeutic potential of these cellular populations, after transplantation into ALS patients and animal models of the disease, in modulating the environment surrounding motor neurons from pro-inflammatory to neuroprotective. We also thoroughly discuss the recent advances made in the field and caveats that need to be overcome for clinical translation of cell therapies aimed at modulating non-cell autonomous events to preserve remaining motor neurons in patients.     

Cortinarius speciosissimus toxins — a preliminary report

Summary Methanolic extracts ofCortinarius speciosissimus yielded a fluorescent compound which was crystallized and shown to be a cyclic polypeptide. The compound, or an analogue, has been found in most members of the genusCortinarius.The authors acknowledge the help of Dr M. Stewart, Department of Biochemistry, Royal Infirmary, Glasgow, for the amino acid analysis. This work was supported by the Scottish Home and Health Department, grant No. H/MRS/S)/C350.     

The amino acid composition of rat bile

Summary The pattern of amino acids in the bile of rats differs from the pattern in the serum of these animals, since bile contains significantly greater amounts of acidic and sulphur-containing amino acids and glycine than serum, while the serum contained more basic amino acids than bile, indicating that secretion of amino acids into bile may involve specific transport processes.Acknowledgments. We thank Mr M. Earlam of Pharmaceuticals Division, Imperial Chemical Industries Limited for the aminoacid analyses, and Dr J. S. Morley for helpful discussion. URF gratefully acknowledges a grant (FO 73/2) from the Deutsche Forschungsgemeinschaft, Bonn-Bad Godesberg, Fed. Rep. of Germany. KGW is recipient of a grant from the Scottish Hospital Research Endowments Trust.     

Genetische und biochemische Untersuchungen über die Bedeutung der Amino- und Nucleinsäuren im Ursachengefüge von Neoplasmen (Tumoren und Gallen). Ein Dauermodifikationsbzw. Prädeterminationsphänomen

Summary If certain hybrids ofPlatypoecilus maculatus andXiphophorus helleri (viviparous platyfish and swordtails) are cultivated in diluted sea-water (0.25 or 0.5% salt concentration), the amount of free amino acids increases and the growth of macromelanophores, which normally only form certain black spots at the dorsal fin, is accelerated. Therefore melanomas arise. After changing salt water for fresh water, growth of macromelanophores remains accelerated. The same effect is seen in progenies which have been in an oocytic or embryonic state, when their mother was cultivated for some weeks in diluted sea-water (Figures 1 and 2). A similar effect has been observed in plant galls produced byPhylloxera (Viteus vitifolii) in the genusVitis. When the parasite, which injects amino acids into the tissue of the host in order to produce galls, is removed, gall formation goes on slowly for some days. It is shown that a few hours after injection of amino acids—before formation of neoplasms begins—the concentration of ribonucleic acids is increased. RNS-(and therefore protein-)biosynthesis in neoplasms may be dependent on concentration of free amino acids. A simple concept for the formation of neoplasms in hybrids of platyfish and galls ofPhylloxera is discussed.

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Den Herren Prof. Dr.G. de Lattin (Zoologisches Institut der Universität des Saarlandes) und Prof. Dr.B. Husfeld (Forschungsinstitut für Rebenzüchtung Geilweilerhof/Pfalz) sind wir für die Förderung dieser Arbeit zu grösstem Dank verpflichtet. Herrn Prof. Dr.C. Kosswig (Zoologisches Institut der Universität Hamburg) verdanken wir viele Anregungen und die freundliche Überlassung von ingezüchteten Stämmen für unsere Fischkreuzungen. — Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Purification of 20α-hydroxysteroid dehydrogenase from human erythrocytes

Summary 20-hydroxysteroid dehydrogenase was found in the supernatant after hemolysis of human erythrocytes, and the enzyme was isolated from the membrane-free hemolysate on a DEAE-cellulose column. The NADPH-generating system was essential for the reaction.Acknowledgments. We wish to thank Dr A. Tomoda, Department of Biochemistry, Kanazawa University School of Medicine, and Dr T. Yubisui, Department of Biochemistry, Medical College of Oita, for their advice and useful discussions.     

The multifaceted role of glial cells in amyotrophic lateral sclerosis

Despite indisputable progress in the molecular and genetic aspects of amyotrophic lateral sclerosis (ALS), a mechanistic comprehension of the neurodegenerative processes typical of this disorder is still missing and no effective cures to halt the progression of this pathology have yet been developed. Therefore, it seems that a substantial improvement of the outcome of ALS treatments may depend on a better understanding of the molecular mechanisms underlying neuronal pathology and survival as well as on the establishment of novel etiological therapeutic strategies. Noteworthy, a convergence of recent data from multiple studies suggests that, in cellular and animal models of ALS, a complex pathological interplay subsists between motor neurons and their non-neuronal neighbours, particularly glial cells. These observations not only have drawn attention to the physiopathological changes glial cells undergo during ALS progression, but they have moved the focus of the investigations from intrinsic defects and weakening of motor neurons to glia–neuron interactions. In this review, we summarize the growing body of evidence supporting the concept that different glial populations are critically involved in the dreadful chain of events leading to motor neuron sufferance and death in various forms of ALS. The outlined observations strongly suggest that glial cells can be the targets for novel therapeutic interventions in ALS.     

Research advances in gene therapy approaches for the treatment of amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease of motor neurons that causes progressive muscle weakness, paralysis, and premature death. No effective therapy is available. Research in the motor neuron field continues to grow, and recent breakthroughs have demonstrated the possibility of completely achieving rescue in animal models of spinal muscular atrophy, a genetic motor neuron disease. With adeno-associated virus (AAV) vectors, gene transfer can be achieved with systemic non-invasive injection and minimal toxicity. In the context of this success, we review gene therapy approaches for ALS, considering what has been done and the possible future directions for effective application of the latest generation of vectors for clinical translation. We focus on recent developments in the areas of RNA/antisense-mediated silencing of specific ALS causative genes like superoxide dismutase-1 and other molecular pathogenetic targets, as well as the administration of neuroprotective factors with viral vectors. We argue that gene therapy offers new opportunities to open the path for clinical progress in treating ALS.     

Metallothionein induced in the frogXenopus laevis

Summary A low molecular weight cadmium- and copper-binding protein, induced in the liver of the frog,Xenopus laevis, by the administration of cadmium, was shown to consist of a single isoprotein and was characterized as an amphibian metallothionein based on its high cysteine and metal content, its low molecular weight, and the lack of aromatic amino acids.The authors thank Dr K. Kubota for his encouragement.     

Therapeutic neuroprotective agents for amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a fatal chronic neurodegenerative disease whose hallmark is proteinaceous, ubiquitinated, cytoplasmic inclusions in motor neurons and surrounding cells. Multiple mechanisms proposed as responsible for ALS pathogenesis include dysfunction of protein degradation, glutamate excitotoxicity, mitochondrial dysfunction, apoptosis, oxidative stress, and inflammation. It is therefore essential to gain a better understanding of the underlying disease etiology and search for neuroprotective agents that might delay disease onset, slow progression, prolong survival, and ultimately reduce the burden of disease. Because riluzole, the only Food and Drug Administration (FDA)-approved treatment, prolongs the ALS patient’s life by only 3 months, new therapeutic agents are urgently needed. In this review, we focus on studies of various small pharmacological compounds targeting the proposed pathogenic mechanisms of ALS and discuss their impact on disease progression.     

Extracellular wildtype and mutant SOD1 induces ER–Golgi pathology characteristic of amyotrophic lateral sclerosis in neuronal cells

Amyotrophic lateral sclerosis (ALS) is a fatal and rapidly progressing neurodegenerative disorder and the majority of ALS is sporadic, where misfolding and aggregation of Cu/Zn-superoxide dismutase (SOD1) is a feature shared with familial mutant-SOD1 cases. ALS is characterized by progressive neurospatial spread of pathology among motor neurons, and recently the transfer of extracellular, aggregated mutant SOD1 between cells was demonstrated in culture. However, there is currently no evidence that uptake of SOD1 into cells initiates neurodegenerative pathways reminiscent of ALS pathology. Similarly, whilst dysfunction to the ER–Golgi compartments is increasingly implicated in the pathogenesis of both sporadic and familial ALS, it remains unclear whether misfolded, wildtype SOD1 triggers ER–Golgi dysfunction. In this study we show that both extracellular, native wildtype and mutant SOD1 are taken up by macropinocytosis into neuronal cells. Hence uptake does not depend on SOD1 mutation or misfolding. We also demonstrate that purified mutant SOD1 added exogenously to neuronal cells inhibits protein transport between the ER–Golgi apparatus, leading to Golgi fragmentation, induction of ER stress and apoptotic cell death. Furthermore, we show that extracellular, aggregated, wildtype SOD1 also induces ER–Golgi pathology similar to mutant SOD1, leading to apoptotic cell death. Hence extracellular misfolded wildtype or mutant SOD1 induce dysfunction to ER–Golgi compartments characteristic of ALS in neuronal cells, implicating extracellular SOD1 in the spread of pathology among motor neurons in both sporadic and familial ALS.     

Stem cell transplantation for amyotrophic lateral sclerosis: therapeutic potential and perspectives on clinical translation

Amyotrophic lateral sclerosis (ALS) is a fatal neurological disease characterized by degeneration of upper and lower motor neurons. There are currently no clinically impactful treatments for this disorder. Death occurs 3–5 years after diagnosis, usually due to respiratory failure. ALS pathogenesis seems to involve several pathological mechanisms (i.e., oxidative stress, inflammation, and loss of the glial neurotrophic support, glutamate toxicity) with different contributions from environmental and genetic factors. This multifaceted combination highlights the concept that an effective therapeutic approach should counteract simultaneously different aspects: stem cell therapies are able to maintain or rescue motor neuron function and modulate toxicity in the central nervous system (CNS) at the same time, eventually representing the most comprehensive therapeutic approach for ALS. To achieve an effective cell-mediated therapy suitable for clinical applications, several issues must be addressed, including the identification of the most performing cell source, a feasible administration protocol, and the definition of therapeutic mechanisms. The method of cell delivery represents a major issue in developing cell-mediated approaches since the cells, to be effective, need to be spread across the CNS, targeting both lower and upper motor neurons. On the other hand, there is the need to define a strategy that could provide a whole distribution without being too invasive or burdened by side effects. Here, we review the recent advances regarding the therapeutic potential of stem cells for ALS with a focus on the minimally invasive strategies that could facilitate an extensive translation to their clinical application.     

Inhibition of steroidogenic activity in the adrenal cortex of rats fed benzene hexachloride (hexachlorocyclohexane)

Summary Feeding various dosages of benzene hexachloride (100, 250, 750 and 1500 ppm) in the diet to weanling male albino rats for 90 days resulted in marked hypertrophy of the adrenals with large, vacuolated cells in the cortex at 750 and 1500 ppm. Accumulation of cholesterol-positive lipids and marked reduction in the activities of steroidogenic enzymes such as ⁵ 3 HSDH, 11 HSDH, G-6-PDH and SDH were seen using histochemical methods in the adrenal cortex of rats fed 750 and 1500 ppm. The results are suggestive of steroidogenic inhibition at 750 and 1500 ppm of dietary BHC while 100 and 250 ppm did not produce any discernible changes in the adrenal cortex.Acknowledgments. The authors thank Dr Muralidhara, H.P. Ramesh and K. Nanjundiah for their help and Prof. H.B. Devaraj Sarkar, Department of Zoology, University of Mysore, for providing us facilities for histochemical work. We are grateful to Sri C.P. Natarajan, for his keen interest in this investigation.     

The non-exponential decay pattern of the weak luminescence from seedling ofCicer arietinum L. stimulated by pulsating electric fields

Our experiments have shown that the response of etiolated seedlings ofCicer arietinum L. to an externally applied pulsating electric field involves a weak luminescence, the intensity of which follows a non-exponential decay pattern.Acknowledgment. It is our pleasure to acknowledge the useful discussions we had with Dr S. K. Srivastava of the Mathematics Department and Dr M. K. Parida of the Physics Department of NEHU, Shillong. We also appreciate the interest shown by Prof. R. K. Mishra, former director of I.S.O.S. & B., in this work.     

Host-plant pyrrolizidine alkaloids inNyctemera annulata Boisduval: Their persistence through the life-cycle and transfer to a parasite

Summary Larvae ofNyctemera annulata Boisduval ingest pyrrolizidine alkaloids fromSenecio spathulatus A. Rich. which subsequently appear in the adult months and their eggs; the acquisition of the alkaloids by a parasite of theN. annulata larvae provides a further illustration of their ability to persist in insects.Acknowledgments. We thank Dr R. Yamdagni of the Chemistry Department, University of Calgary, Canada, and Dr O. Madsen of the Organic Chemistry Department, The Technical University of Denmark, Lyngby, Denmark, for much help with the MS data collection. We are also very grateful to Dr C.C.J. Culvenor, CSIRO Animal Health Lab., Parkville, Victoria, Australia, who most generously provided us with authentic reference specimens of pyrrolizidine alkaloids.     

The reported occurrence of L-canavanine in soya bean,Glycine max

Summary The reported occurrence of L-canavanine in soya bean,Glycine max could not be verified by enzyme treatment of the extracted non-protein amino acids of the seed.This work was supported by a grant from the National Science Foundation (BMS-75-19770) and funds from N.I.H. Biomedical Grant 5-SO5-RR07114-08.The author thanks Dr D. B. Egli of the University of Kentucky for soya bean seeds and Karen Hughes for her technical assistance.     

Effects of tranylcypromine and pargyline on brain tryptamine

Summary Tranylcypromine produces behavioral excitation while pargyline produces depression. Tranylcypromine increased brain tryptophan which led to an accumulation of tryptamine. The levels of tryptamine after tranylcypromine were found to be 3 times those found after pargyline.This work was supported in part by U. S. Public Health Service Grants NS-12759 and AA-2696, State of Illinois Department of Mental Health, the Psychiatric Services Branch, Province of Saskatchewan, and the Medical Research Council of Canada, who have provided continuing financial support.Acknowledgments. We thank Dr B. A. Davis for synthesizing the deuterated internal standards, Dr D. A. Durden for supervising the mass spectrometric analyses, and Mr H. Miyashita, Mr N. F. Binder and Miss E. E. Johnson for skilled technical assistance.     

Nuclear blebs in oocytes of the fishClarias batrachus

Summary Large nuclear blebs have been observed in oocytes of the fish,Clarias batrachus. The bleb, which contains nucleoplasm, is finally extruded from the nucleus. The extruded body is DNase-sensitive, and it implies that some amount of DNA is extruded from the nucleus of these oocytes.Acknowledgments. We are thankful to Dr. R.K. Sharan, Department of Zoology, Patna University, for providing laboratory facilities, and to Dr V.G. Jhingran and Dr P.V. Dehadrai of ICAR for their keen interest in the work.     

Effects of dopamine antagonists on snail locomotion

Summary Dopamine antagonists which affect the ergometrine-sensitive type of dopamine receptors produce characteristic motor disorders in a land snail,Helix pomatia L.Acknowledgment. We are grateful to Dr L. Hiripi for valuable suggestions and Dr I. Varanka for help in photography.