Central nervous system involvement in mycosis fungoides: pre mortem diagnosis by light and electron microsocopic study of the spinal fluid. Involvement of the brain, spinal cord, peripheral nerves, meninges, and cerebrospinal pathways in mycosis fungoides manifests itself, clinically, as an obscure neurologic disturbance, occurring in patients with cutaneous manifestations of that disorder. Neurologic expressions of this particular cutaneous lymphoma are infrequently reported, in spite of the fact that many patients dying in the course of mycosis fungoides have pathologically demonstrable central nervous system, peripheral nervous system, and meningeal involvement. We report here a patient who incurred devastating neurologic illness as one of the initial features of the course of systemic dissemination of mycosis fungoides. Death occurred nine years after the onset of cutaneous illness. In this instance, cerebrospinal fluid cytologic studies established the diagnosis of nervous system involvement. Mycosis fungoides cells shed into the cerebrospinal fluid were studied by special techniques of light and electron microscopy. Brain scan abnormalities were present, which were clinically correlative with foci of brain involvement. The initial response of patient's nervous system illness to therapy was gratifying but unsustained, and she went on to fatal brain and multiple organ involvement with disseminated mycosis fungoides. The techniques of spinal fluid cytologic diagnosis, confirmatory of the etiology of disease in this case, may have practical appications in the diagnosis of otherwise obscure neurologic disorders.


[Saint Louis encephalomyelitis in Hermosillo, Sonora, Mexico. Its epidemiology]. In a warm desertic and irrigated region, with population 250 000 Northwest of Mexico, in Hermosillo, Sonora, along 8 weeks, 51 patients showed the following clinical data: fever, vomiting, neck rigidity, drowsiness and increase of cells and proteins in the CSF. In 12 patients good correlations were found between positive serologic results using SLE antigen by means of HI, CF and NT. Four paired sera showed fourfold increases in titers to SLE by means of HI or CF. With VEE antigen no such increases were found. The cases were scattered geographically, most of them occurred within an urban area, and affected people in 13 localities. The outbreak developed from August to September, 1974, the attack rate was 22/100 000, 69% of the patients being children under 15 years of age. The case fatality rate was 20%. A prosperous chicken industry existed in the region and epizootics were not reported in either domestic or wild animals at that time.


A new model of persistent viral infection with primary demyelination. Theiler's encephalomyelitis virus (TMEV) in mice represents one of the few existing animal models of virus-induced demyelination and may provide further information about the human disease, Multiple Sclerosis. An inbred strain of mice (SJL/j) employed in the present ultrastructural study, develops a life-long neurologic illness characterized by profound spasticity after intracerebral inoculation of TMEV. Anesthetized animals were sacrificed sequentially by total body perfusion with gluteraldehyde from 7 days to 9 months after infection. An intense mononuclear inflammatory infiltrate appeared in the leptomeninges and white matter of spinal cord by 15 days and persisted for as long as one year. These infiltrates contained numerous plasma cells, and perivascular demyelination could be found in almost every 1 micron section. Vesiculation and stripping of myelin by mononuclear cell processes were seen as mechanisms of myelin break-down. However, virions were not found in cells in grey or white matter and the integrity of oligodendrocytes was maintained. At one year, active demyelination was still seen in areas containing naked and remyelinated axons. The relevance of these ultrastructural findings to postulated mechanisms of virus-induced demyelination will be discussed.


A neurophysiological basis of developmental dyslexia. Recent advances in the understanding of the asymmetrical functions of the cerebral hemispheres provide new insight to the mechanisms underlying the developmental language disorders. It is probable that learning to read depends in part upon the ability to establish an association between a seen object (customarily perceived within the right hemisphere) and a verbal symbol (mediated by the left). It is possible that as the reading skill develops, verbal symbol recognition may also become a left hemisphere function. A complex interaction between the two developing hemispheres is involved. A clue to the nature of this problem is that developmental dyslexia is six times more common in boys than in girls. Studies defining differences in cerebral dominance in boys and girls will be reviewed in this context.


Nervous system affections caused by the mumps virus. Hippocrates probably first described mumps parotitis, but not until 1758 was affection of the central nervous system reported in this disease. Mumps meningitis, encephalitis, myelitis, polyradiculitis and cranial neuritis are now well known, and may occur without clinical parotitis. Meningitis occurs most commonly, encephalitis, cranial neuritis and polyradiculitis less often, and myelitis rarely. They may present individually or in combination. A patient is described who first developed acute mumps meningoencephalitis, without clinical parotitis. Transverse myelitis occurred two weeks later, and finally optic neuritis ten days following the myelitis. Slow but complete recovery followed. Attention is directed to the various ways mumps can affect the nervous system. This virus should always be considered among possible etiologic agents causing such neurologic syndromes.


Virological and immunological studies in experimental SSPE. Subacute sclerosing panencephalitis (SSPE) is a progressive, fatal inclusion cell encephalitis of children and adolescents caused by persistent measles virus within the central nervous system (CNS). Because studies in man have failed to elucidate the pathogenesis of this condition, animal studies are necessary. Persistent infection of the hamster CNS can be achieved with a hamster adapted SSPE agent. Animals inoculated intracerebrally with this virus raise antibodies to all known antigens of measles virus and some display clinical signs and pathological changes similar to those noted in human SSPE. Persistent CNS infection occurs only if the hamster is inoculated at a critical age (18 to 25 days of life) or if adults are given transient immunosuppression during acute infection. The biological behavior of the virus isolated from hamster CNS appears to change from a complete to a defective state coincidents with the appearance of serum antibodies to measles virus. Adult hamsters from whom the thymus was removed in the newborn period develop a subacute, uniformly fatal infection when exposed to the SSPE agent. These studies suggest that SSPE may develop in man when measles virus invades the immature CNS at a critical age or when the immune system is uncompletely developed or is inhibited. The finding that transient immunosuppression allows development of persistent CNS infection in adults suggest that immunological malfunction is the significant factor. Of interest, antibody appears to alter viral behavior to a defective, intracellular state thus enhancing viral survival in the host. Once a defective, CNS infection is achieved, lack of, or inhibition of the host cellular immunes response allows it to persist. Methods of therapy in light of these findings will be discussed.


Immunological response of subacute sclerosing panencephalitis patients to measles virus. The immunological responses of patients with clinical signs of SSPE were examined by the direct migration inhibition, complement-mediated antibody cytoxicity and the lymphocyte-mediated cytotoxicity assay techniques. Peripheral lymphocytes migrated similarly to controls when exposed to measles antigen. Lymphocytes from SSPE patients responded to a SSPE strain of measles infected cells in a manner similar to lymphocytes from controls. A factor which interfered with the expected activity of sensitized lymphocytes to measles virus was detected in some cerebral spinal fluid obtained from SSPE patients. The blocking activity varied from 100% in 4 of 20 patients with SSPE to no blocking in 2 of the SSPE patients. The occurrence of blocking factor in SSPE patients could not be related to sex or age of the patient, serum of CSF antibody levels, and duration of disease or outcome.


Leukapheresis in multiple sclerosis.

Leukapheresis in multiple sclerosis. Since there is no satisfactory animal model for multiple sclerosis (MS), a need exists to study various tissues of the affected patient himself. Leukapheresis is a relatively non-invasive technique for obtaining large numbers of leucocytes from patients and control patients. Using ACD-B solution as anticoagulant, the procedure can process 3 500 ml. of blood in about 2 hours, and yield an average of 6.7 billion lymphocytes. The circulating blood count and differential leucocyte count are not changed significantly at the end of the procedure, although the platelet count drops slightly. Leukapheresis has been performed 6 times in four MS patients representing all stages of the illness. The data indicate that the procedure, as performed, is safe in MS patients and do not adversely alter the course of the illness. Leukapheresis is not suggested for therapy at this time. However, if further study suggests that circulating lymphocytes in the MS patient have a cytolytic function in the central nervous system (CNS), the procedure might merit a trial in a controlled treatment program. At present, leukapheresis will probably be most useful as a research tool, to aid study of the role of the lymphocyte in the pathogenesis of MS.


[Viral meningoencephalitis: late development]. Two years ago, we performed a precocious evolutive EEG study of a specimen not selected from the virus ME to ECHO 4 burst (cepa prima) late 1972 to early 1973 (6). Some time between the appearance of the first symptoms and the 8 weeks following, alterations of cerebral electrogenesis became manifest in this study. These alterations disappeared or dimished in significance in the more tardy outlines: focalization tendency, tendency to change from "suffering" to "irritative" aspect. During this year, the test has been repeated on 82% of these patients, plus at the same time a minimum clinical survey. Until now, the results are the following. There are no clinical signs of the CNS in any case. School progress has not been affected, except moderately, for two patients. The EEG has become totally normal in more than 2/3 of the cases. Persistent EEG net signs of minimum "suffering" in three cases. There are "irritative" generalized signs in only 2 patients and "irritative" certified focal signs in 6 patients. Other EEG manifestations are of doubtful interpretation due to the age of the patients.


Experimental subacute sclerosing panencephalitis in the hamster: ultrastructure of the chronic disease. Central nervous system (CNS) lesions were studied from weanling hamsters inoculated with the HBS strain of subacute sclerosing panencephalitis (SSPE) virus. The animals showed clincial signs of acute encephalitis between 8 and 18 days post-inoculation (PI), but all survivors were clinically recovered by day 21 PI. Nevertheless, 13 of 14 hamster brains examined by light and electron microscopy between days 21 and 59 PI had chronic lesions which contained morphologic evidence of persistent viral infection. The lesions developed preferentially in the subependymal areas of the lateral and fourth ventricles and involved degeneration of ependyma with subsequent damage to adjacent parenchyma. All CNS cell types were involved in degeneration. Viral inclusions occurred in both CNS parenchymal cells and in inflammatory cells. Giant cells were particularly common. No budding virus was seen in chronically infected animals, a finding in accord with previous studies. Demyelination was a common constituent of most lesions. It occurred in the presence of inflammatory cells and macrophages, and in later lesions, some remyelination was seen. It is suggested that the damage to myelin is a secondary phenomenon and is not a cellular immune reaction. The possible reasons underlying the latent nature of the virus and the similarities between this condition, canine distemper encephalomyelitis and human SSPE are discussed. It is concluded that the experimental chronic disease is a valid model for the study of human SSPE and may have usefulness in the understanding of other chronic CNS conditions of man, e.g. multiple sclerosis. Additional Key Words: Latent infection; Paramyxovirus; Slow Viruses; Demyelination; Inmmunologic defects; Multiple Sclerosis.


[What, to whom and why are we teaching medicine]. The teaching of neurology as an entity within the concept of teaching in medicine is analized through the following questions: what are we teaching? To whom are we teaching? and why are we teaching? The first question concerns the pedagogical, sociological and psychological patterns used to elaborate locally-arranged goals for the creation of criteria and conduct. The second establishes the relationship between the influence of the technical-industrial adult generation on the present university generation and the reappraisal of human values through new pedagogic techniques. The third establishes the supply and demand process of assistant teaching and research physicians; it also dwells on the influence of the residency programmes and research trends, and their repercussion on the methodology of the medical curriculum.


[Pedagogical experience and learning models in neurologic semiology]. Programmed teaching of neurologic semiology, charted in agreement to subsequent learning in clinical neurology, is conceived. The basis of the pedagogic system consists of respecting the systems that bind physiologically among themselves. The goal is the creation of areas of comprehension, dramatizing common functions and signology. To know "all" is of less validity than to posses an engram prepared by key information. The anatomic physiology memory, previous to the course and the weekly partial evaluation, is considered useful. The problematic of the competitive test is analyzed and its negative and positive criterions are exposed. Anatomy, physiology, semiology and clinical neurology spun together are estimated to favor pedagogic goals redundantly.


[Formative evaluation of education].

[Formative evaluation of education]. The formative evaluation tries to obtain, analyze and furnish information over the planning and development of a course, which is brought about through a continuous performance of the students during the course. This method allows for the adoption of corrective measures with reference to the course (presentation of other learning opportunities, preparation of instructive materials, revision of curriculum, etc.) and with reference to the student (studies prescribed). The formative evaluation may be fulfilled by means of repeated tests which the student himself will correct and, subsequently, will discuss with the professor; it can also be processed by means of the students self-evaluation, interacting with computer terminals. CLATES/Rio presents its experience employing minicomputers to allow for self-evaluation in bioscience courses.


[Experiences in neurology education in Cuba]. The state of medicine and neurology in the past years, since 1962, is described. The initial personnel with which neurology and neurosurgery began working, the manner in which the group was formed and the subsequent formation of specialists in this area are commented. We describe the hours and subjects studied by the residents and the modifications that the program has experienced since 1962, until 1971, in which a program of philosophy, social and political lectures, combined with the specialization classes, exists. An evaluation system, created by the group and number of specialists formed since its initiation, also exists. Subsequent to their formation, physicians integrate neurologist and neurosurgeon teams and are situated in areas of provinces which cover a population of 500 000 to 1 500 000 inhabitants.


Artistic production in dyslectic children. In the study of children with language problems, particularly in reading and writing, it has been observed that some have an outstanding ability to produce artistic pictures and objects. These productions are perceptive, well organized and generally contain much action. Despite their pictorial skill these patients may have only a rudimentary use of coded symbolic graphic forms. Others display moderate ability in reading and writing. These patients frequently have the disorganized overacctive behavior and the motor clumsiness that is so common in the dyslectic child; some, however, are biologically effective. From this material we entertain the hypothesis that picture (artistic) productions are generated by the sub-dominant cerebral hemisphere, and that this function is quite distinct from the coded graphic operations resident in the dominant hemisphere. If this hypothesis is correct, it would seem socially benefical to allow these patients to develop their unique artistic ability to its full capacity, and not to overemphasize the correction of the disturbed coded symbol operations in remedial training.


Speech and swallowing evaluation in the differential diagnosis of neurologic disease. Examination of the speech mechanism (the vocal tract) frequently provides information about certain neurologic lesions. The speech, swallowing and respiratory patterns of patients with cerebellar, upper motor neuron, lower motor neuron and extra-pyramidal lesions (parkinsonism) were studied, 10 of each category. The patient's lingual, labial and velar movement was examined cineradiographically during production of four short speech samples and the oral, through esophageal structures, during chewing and swallowing. Videotape, with simultaneous voice recording, and 16 mm. film were used. Oral, pharyngeal and esophageal transit times during swallowing were determined. These quantified tests reveal distinctive differences in vocal tract physiology between patient groups. The specific physiologic patterns characteristic of each neurological lesion were discussed and illustrated with films. Each patient group in this study exhibited a distinctive profile distinguishing it from every other. The results also indicate that if only a portion of these studies was completed, the disorder might be erroneously classified. The findings emphasize the need for complete and detailed physiologic studies of the speech and swallowing of patients with neurologic lesions, as an aid to accurate diagnosis.


A different form of "pure agraphia": syntactic writing errors in a patients with motor speech and movement disorders. Three patients are described who displayed syntactic writing errors in combination with a motor speech disturbance and impaired motor limb function. Two of the patients had bulbar amyotrophic lateral sclerosis (ALS). Agraphia appeared when verbal communication was no longer possible. Autopsy in one patient disclosed only lesions consistent with ALS. The third patient had palilalia and chorea and although not aphasic, his written language showed persistent syntactic errors. We hypothesize that the agraphia in these patients occurred because of the combination of disordered feedback from the motor speech apparatus and limbs.


[Nature of repetition disorders in conduction aphasia]. The skeptic over the existence of this important clinical entity, are now very few. Anatomo-clinical studies and the analysis of altered cognitive procedures suggest, however, that we are dealing with a heterogeneous group. Two models of conduction apahasias are presented, in which the mechanism of the altered repetition, as well as the probable localization of the lesion, differ considerably. One has a lesion in the posterior parietal region of the left hemisphere and it exemplifies an acoustic verbal memory deficit (short term memory). The other points at a functional disconnection between the decodification and verbal processes and those of expression. In this case, we are dealing with a left temporal lesion in a left-handed individual. The analysis of our material and of the clinical literature upholds the idea of a plurality of the altered mechanisms in the verbal repetition and suggest that the conduction aphasia is a syndrome.


Dynamic characteristics of central auditory neurons and their relation to selective-combination mechanisms of organization of brain functions. Unit responses of the torus and caudal neostriatum of hens to stimuli of differing ecological significance (pure tones, white noise, species-specific stimuli) were investigated. The range of frequencies receivable by central auditory neurons was shown to correspond to the frequency composition of the stimuli emitted by the animals. Neurons selectively responding to species-specific stimuli (song, alarm signals) were found in the forebrain. The specific character of the functional organization at different levels of the auditory system is examined in the light of the selective-combination principle of stimulus integration, which may lie at the basis of the mechanism of both inborn and acquired memory.


Afferent flow patterns in a cat cutaneous nerve during painful and painless mechanical stimulation of the skin. Quantitative characteristics of afferent flows coding information from a number of receptors were obtained by the gliding impulses method. The frequency spectrum of activity in a cutaneous nerve, the relative numbers of active Abeta, Adelta, and C fibers and their distribution by impulse transition frequency during stimulation of the cat's skin with pins and needles were determined. The afferent flow recorded in the nerve during pricking of the skin is characterized by high density, due to the number of excited fibers and the frequency of activity in them. The higher density of the afferent flow during the application of a painful than of a painless stimulus is mainly due to activity in C fibers. Unmyelinated fibers subjected to the action of the same stimulus and of chemically active substances liberated from the cells during tissue injury are excited directly and generate high-frequency spikes which increase the flow density in the nerve. The number of active myelinated fibers and the spike frequency during the action of a painful stimulus are only a little greater than the corresponding characteristics of the afferent discharge during painless stimulation.


Selective action of decimeter waves on central brain structures. To study the sensitivity of some central brain structures to the action of an electromagnetic field of decimeter waves (EMF of DW) a dynamic investigation of single unit activity was undertaken. Neurons of the hippocampus and hypothalamus with an excitatory type of response were found to have the greatest reactivity to EMF of DW. Neurons of the mesencephalic reticular formation and of the specific nuclei of the thalamus had mostly an inhibitory type of response. The number of neurons responding actively to EMF of DW and the number of inactive neurons varied depending on the energy density of the EMF.


Saccadic velocities in multiple sclerosis and myasthenia gravis. Horizontal saccadic eye movement velocities have been measured in patients with multiple sclerosis and myasthenia gravis using a computerised electrooculographic technique. The ability to detect subclinical abnormalities of eye movement with this technique indicates that it is of diagnostic value in patients with suspected multiple sclerosis who present with only a single symptomatic lesion in the central nervous system. The technique is also of value in the diagnosis of myasthenia gravis as it provides a means of quantifying the response of the extra-ocular muscles to anticholinesterase preparations.


Electrophysiological and immunological studies in optic neuritis. Serial clinical and electrophysiological examinations and immunological investigations have been performed on 34 patients with primary optic neuritis. In 8 patients, the latency of visual evoked responses was normal after the resolution of clinical symptoms, and in a further 5 patients improvement in VER latency occurred although the response remained abnormal. In 17 patients, VER's abnormal and did not alter over the period of clinical recovery; in 4 patients there was a progressive increase in the latency of the VER's through both eyes although a satisfactory clinical recovery occurred. The frequency of histocompatibility antigens was similar in the group of ON patients to that in the normal population. Immunological function was normal in patients with ON as assessed by levels of immunoglobulins. T and B lymphocyte numbers and by lymphocyte blastogenesis in response to PHA and myelin basic protein. Three patients have developed multiple sclerosis, all of whom had either HLA-A3 or -B7 antigens; the clinical and electrophysiological pattern of recovery was different in each case.


Peroneal muscular atrophy with autosomal dominant inheritance. Nerve conduction studies were performed on 45 patients in 17 kinships with autosomal dominant Charcot-Marie-Tooth disease. Sural nerve biopsies were performed on 20 patients. The mean motor conduction velocities in median, ulnar and lateral popliteal nerves were 27.5m/sec, 25.9m/sec and 26.3m/sec respectively. Sensory conduction was impaired in all cases. The fastest motor conduction velocities were found in 5 members of 1 kinship who were found on sural nerve biopsy to have the neuronal form of the disease in which there was a reduction in density of large diameter myelinated fibres. In this group segmental demyelination was not a prominent feature in teased fibre preparations and onion bulb formations were not seen on electron microscopy. In the other patients conduction velocities were more severely slowed and hypertrophic changes and segmental demyelination were found in sural nerve biopsies.


Resting tremor in alcoholic brain disease. Two patients with signs of alcoholic brain disease of the Wernicke-Korsakoff type were found to have an unusual movement disorder manifested basically as a resting tremor. Electrographic recording showed that the tremor wave comprised multiple elements, though the rate of the basic tremor was stable. Though the tremor was qualitatively similar to documented rubral tremor, a mesencephalic lesion could be defined in only 1 patient.


Factors influencing plasma carbamazepine concentrations. Steady-state plasma carbamazepine levels were correlated with carbamazepine dose, expressed on a body weight basis, in 217 patients (some of whom were taking other anticonvulsants). Although there was a linear relation between plasma carbamazepine level and drug dose in the whole population studied, no such relation was found for patients taking carbamazepine alone. The factors responsible for this difference could not be identified in full, but interactions between carbamazepine and phenytoin were partly responsible.


Spinal and cortical evoked potentials in multiple sclerosis. Spinal and cortical sensory evoked potentials following stimulation of median and lateral popliteal nerves were performed in patients with definite multiple sclerosis in order to assess the usefulness of these procedures in the diagnosis and subsequent evaluation of multiple sclerosis. Abnormal SEP's were detected in 69% of patients. Abnormal spinal evoked potentials following median nerve stimulation were detected in 21 of 40 (52%) patients, and in a further 7 patients abnormalities were detected in cortical SEP's or in spinal SEP's when stimulating the lateral popliteal nerves. Close correlation was observed between the frequency of abnormal sensory signs or symptoms and the presence of electrophysiological abnormalities. When progress studies were performed a gradual deterioration was demonstrated in one or more SEP's of 12 of 24 patients.


Electrophysiological and computerised tomography findings in multiple sclerosis: a comparative study. 102 patients with multiple sclerosis (MS) have been investigated by one or more of the following techniques: visual evoked potentials, cervical and cortical somatosensory evoked potentials, horizontal saccadic eye movement velocities, and computerised axial tomography of the cranium and orbits. It has been found that with each technique it is possible to detect abnormalities, which may be subclinical, in a significant proportion of patients. The proportion with abnormal results was greater in patients studied by more than 1 technique and was greatest in those patients who were studied by each of the 5 techniques. It is concluded that the techniques used have a complementary role in the investigation of patients suspected of having MS.


Computerised tomography findings in multiple sclerosis and Schilder's disease. Computerised tomography of the brain was performed in 95 patients with multiple sclerosis and in a single patient with Schilder's disease. CT of the orbits was also carried out in 53 of these cases to examine the optic nerves. In 51% of the multiple sclerosis cases low density areas compatible with plaques of demyelination were found in the white matter of the cerebral hemispheres, and less frequently in the brain-stem. A much larger area of reduced intensity was found in the occipital white matter in the patient with Schilder's disease. Varying degrees of central and/or cortical cerebral atrophy were found in 45% of the multiple sclerosis patients, the more severe degrees being found in patients with long-standing disease. Serial observations in 15 patients who were examined on more than one occasion are also presented. Small low density areas were identified in one or both optic nerves in 52% of the multiple sclerosis cases. While these may represent demyelinating lesions their significance is uncertain.


Computerised tomography of the cranium in patients with epilepsy: a preliminary report. The findings are presented in 366 patients with seizures who were studied by computerised tomography of the cranium. The relative frequency of normal scans, cerebral tumours, atrophy and other pathological lesions is indicated. A more detailed analysis of the group of patients with atrophy was carried out. The frequency and severity of atrophy correlated significantly with age but not with length of seizure history. However, the more marked degrees of atrophy were found among patients with long-standing or poorly controlled epilepsy, an observation which has important implications with regard to the adequacy of seizure control in epileptic patients.


Distal chronic spinal muscular atrophy involving the hands. Six patients with a 10 to 30 year history of slowly progressive wasting and weakness of the small muscles of the hands have been studied. In all patients the abnormal neurological signs were largely confined to the hands and there was no clinical nor radiological evidence of peripheral nerve, nerve root or central nervous system disease. Detailed electrophysiological studies established that the muscle wasting was due to chronic partial denervation. Motor and sensory conduction studies were normal. These patients represent a benign form of chronic spinal muscular atrophy largely localised to the small muscles of the hand.


An investigation into reading epilepsy.

An investigation into reading epilepsy. In summary, our main findings were firstly, that faulty visuolingual processing at the level of Brain's visual word schemas, with its intimate connection with the verbal memory pool, is the abnormality that is essential for initiation of spike activity in reading epilepsy. Secondly, visual input is not essential in production of reading epilepsy. Thirdly, we have demonstrated a subconscious voluntary control mechanism operating in our patient. Lastly, we have illustrated alternatives to drug therapy, which, in our patient and in other cases, has not been shown of great value.


Effects of neostigmine and pyridostigmine at the neuromuscular junction. The effect of subcutaneous administration of pyridostigmine or neostigmine for 7 to 15 days on neuromuscular transmission has been studied in the rat phrenic nerve-hemidiaphragm preparation. The quantal contents of end-plate potentials at different stimulus rates and the amplitude and frequency of miniature end-plate potentials were compared with those of untreated controls. The rate of release of acetylcholine quanta at high stimulus rates, and the frequency of miniature end-plate potentials, were reduced by pretreatment with both pyridostigmine and neostigmine. Presynaptic effects differed in that the number of quanta released by each nerve impulse at a stimulus rate of 1/sec was not altered by pyridostigmine, but was reduced to 52% of normal by neostigmine. The amplitude of miniature end-plate potentials was reduced to 81% by pretreatment with neostigmine and to 54% by pretreatment with pyridostigmine. The cause appears to be a reduction in the number of acetylcholine receptor sites as a result of disorganisation of the postsynaptic muscle membrane, which may contribute to the muscular weakness associated with the long term use of anticholinesterase agents.


Motor neuron disease in Australia (State of New South Wales). The case histories of 70 patients with typical motor neuron disease (MND) have been analysed to ascertain the natural history of the disease in Australia. The place of birth and residence during life of each patient has been sought to determine whether there is any 'clustering' of patients in the State of New South Wales. The mean age of onset was 55 years and mean duration of life from onset was 3 years 5 months. These figures are comparable with those from the USA and UK and Japan although the onset in Australia was later and the duration longer. The case history of 1 patient who is still living 38 years after the onset of the disease is described as there is no longer survival time recorded in the literature to our knowledge. Attention is drawn to muscle aches and cramps mentioned spontaneously by 27 of the 70 patients. These symptoms occurred at the onset of MND and often correlated with the site of onset of weakness. Weight loss averaged 1.6lb (0.7kg) per month in those patients in whom weight was recorded. The CSF protein was greater than 45mg/100ml, ranging up to 110mg/100ml, in 9 of 37 patients. The place of residence of patients bore no relation to known areas of increased manganese in the soil, and there was no evidence of 'clustering' of cases.


The carpal tunnel syndrome: a clinical and electrophysiological study of 250 patients. Clinical and electrophysiological studies were performed on 250 patients with carpal tunnel syndrome. Acroparaesthesia was often referred to all digits (64%) and proximal pain was not uncommon (46%). The male patients as a group differed from the female in that nocturnal acroparaesthesia was less frequent and a causal of predisposing factor was identifiable in the majority (76%). The most common associated condition in the females was trigger finger (11%). An associated ulnar nerve lesion was rare. Delay in diagnosis was attributable to cultural factors or lack of awareness of the occurrence of diffuse acroparaesthesia and proximal pain in the syndrome. Sensory signs were commoner (85%) than motor (53%). The most useful clinical finding was digital sensory 'splitting' (ring finger) which was present in 58% of hands and clearly distinguished the condition from a radiculopathy, myelopathy or a diffuse polyneuropathy. In general, the diagnostic yield of any test depends on the duration of the disease. Of the various electrodiagnostic tests employing surface electrodes, the most sensitive was the comparison of the median (index finger) to ulnar (little finger) sensory amplitude. This was abnormal in 73% of hands and was found to be particularly useful in patients with a probable background of polyneuropathy or suspected cervical rib syndrome. Slowing of sensory conduction was demonstrable in 67% of hands, decreased sensory amplitude in 44% and prolonged motor latency 35 to 32%. A combination of the various electrodiagnostic tests yielded a positive result in 91%. Decompression resulted in complete recovery in 85% and some improvement in all the remaining patients. The outcome was adversely affected by the presence of several neurological deficits but apparently not by the presence of a coexisting disease.


Ocular motor involvement in post-infective polyneuropathy. Ocular motor involvement is uncommon in post-infective polyneuropathy. Details of the evolution and recovery of the ophthalmoplegia are presented in 11 patients in whom ocular motor involvement was a major or notable feature of the illness. Follow-up examinations after intervals of up to 6 years in 10 of the 11 patients showed no residual ocular motor impairment. Saccadic eye movement studies carried out in 6 cases at the time of follow-up showed no abnormality in 2 cases while in the other 4 cases there was mild reduction of saccadic velocities and/or increased saccade reaction times with saccade inaccuracy. In spite of the variability of the ophthalmoplegia and of the remainder of the clinical syndrome in these cases, we conclude that they all represent variants of the classical form of the Guillain-Barre syndrome.


Syndrome of ophthalmoplegia, ataxia and areflexia. The clinical details and results of some laboratory investigations are described in 4 patients who initially presented with severe external ophthalmoplegia, ataxia and areflexia. In 3 of these patients paresis of the limbs was restricted and minimal as in the syndrome first described by Fisher (1956). The fourth patient initially presented with similar symptoms but his illness progressed to a more typical form of acute idiopathic polyneuropathy, confirming Fisher's (1956) contention that this syndrome is an unusual variant of acute idiopathic polyneuritis.


Post-irradiation extracranial cerebrovascular disease. It would seem that most reports of post-irradiation extracranial cerebrovascular disease concern the long-term effects of this form of therapy. Three such cases are described here. The features peculiar to this form of disease are its severity and very considerable extent. Radiologists and vascular surgeons contribute heavily to its diagnosis. Whilst it would appear that rarely in early cases a type of vascular lesion peculiar to radiation damage has been encountered, for the most part reports have emphasised the presence of very extensive and accelerated premature vascular aging and atherosclerosis as a fundamental lesion probably involving all laminae of the vessel. In addition extensive perivascular fibrosis and overlying post-irradiation skin atrophy can be seen. Therefore in patients presenting with cerebrovascular pathology even years after receiving radiation therapy, the neurologist and vascular surgeon must be prepared to recognise that extensive damage may underlie the patients' symptoms and that investigation and surgical repair may have to be undertaken forthwith if at all practicable.


Intermittent hydrocephalus due to cysts of the septum pellucidum: a study of three cases. Three male patients with posturally induced, intermittent hydrocephalus were found on pneumoencephalography to have cavum septi pellucidi and cavum vergae. The clinical presentation resembled that of colloid cyst of the third ventricle. Remission followed excision of the septal walls in 2 cases, but relapse of symptoms occurred when the caves reformed. It is postulated that intermittent obstruction to the lateral ventricular outflow by the cavum septum pellucidum and cavum vergae was the basis of the symptomatology. Transcaval shunting may prove to be the simplest reliable form of treatment.


[Forensic-psychological analysis of attempted suicides in toxicomaniacs]. The authors examined 70 toxicomaniacs incl. 30 who attempted suicide at least one. They analyzed the motivation and causes which made the adolescent attempt suicide. The drastic manner of attempted suicide-cutting veins of the forearm and wrist predominated ans was recored in 56.6%. 83.3% attempted suicide during acute intoxication. Mild cerebral dysfunction was observed in 43.3% of those attempting suicide. Defective behavior was recorded in the case-history of 63.3%. In 80% of those attempting suicide the family and social environment was unfavourable. Dependence on psychotropic drugs for more than one year was found in 80% and polytoxicomania in 90% of the cases. Volatile psychotropic substances cause considerable deterioration of already existing changes of personality of encephalopathic or psychopathic origin; they act probably as "hasteners" or "catalyzers" and thus condition the motivation for suicide. This fact is important in particular for forensic psychology and expert opinions on cases of completed suicide.


[Modern concepts and methods in forensic medicine education]. The authors deal with the modernisation of contents of lectures and practicals in forensic medicine, their politcal and educational effect, the position within the framework of medical sciences and its incorporation in the teaching process and the use of developed technical advances in contemporary teaching.


[Routine chemical-toxicological examination in fata poisoning through organophosphates]. The authors describe in their work methods of isolation of organophosphate insecticides from biological material--the isolation of the effective substance proper as well as of the solvent. They detect organophosphates by means of chromatography on a thin leyer and in one instance also by gas chromatography. In conjunction with chromatography on a thin leyer they discuss methods of detection according to the chemical structure of the substances searched for and submit their own modification of detection based on inhibition of acetylcholine esterase. They detect the solvent in insecticides in biological materials by means of gas chromatography.


[Attempt at a new look on the problems of traffic accidents type pedestrian--car]. In view of the complexity of investigation of the ever rising number of traffic accidents the authors draw attention to the need of a narrower specialization of experts in the branch of forensic medicine and on the desirability of close collaboration and technical specialist when elucidating mechanisms of traffic injuries. In one of the most pretentious sections, in traffic accidents of the type pedestrian--car, they want to attempt an interdisciplinary study the purpose of which is to obtain certain basic data for expert evaluation of the mechanism of fatal injuries of pedestrians, and a basis for assessing speed limits at sites of increased danger of this type of accidents. As source of information they will use in particullar: the protocol of the accident incl. the map and photodocumentation, a questionnaire prepared for the investigating authorities, the results of necropsy incl. laboratory examinations, and if available, conclusions of expert opinions from other special branches.


[Finding of analgesic components and their metabolites in doping control]. In doping controls it is important to exclude essential substances that are not in the list of forbidden drugs. Similarly it is important as well for these drugs to be proved in analgesic mixture abuse. An example being the proof of amidopyrine and its metabolites, further on p-phenetidine as a metabolie of phenacetin in alkaline urine extracts. Results are given in using thin-layer chromatography, gas chromatography and the identification by means of gas chromatography/mass spectrometry.


[Determination of the C3 component in relation to paternity suits]. The C3 component which is a part of the complement appears to be of significance in assessing fatherhood in paternity causes. Its occurrence follow hereditary laws, in homozygotes the phenotype is represented by one, in heterozypotes by two zones designated as S, F and SF. The demonstration depends on the quality of the serum and the laboratory technique used. The same applies to the reliability of the readings.


Large-scale production of human interferon in lymphoblastoid cells. The methodology for optimum Namalva cell growth and interferon production has been developed. Maximum yields of interferon were obtained following dilution of a saturation density suspension culture induced with NDV.


Conventional assay systems for interferon. The continuous appearance in the literature of new and modified assays for interferon indicates the lack of a universally acceptable assay. Recent reports show a tendency for simple, rapid, and economical assays with capabilities for the assay of interferons from many animal species. The EMC HA yield reduction assay, the neuraminidase yield reduction method, and the radiochemical assay have many desirable features. These assays have the potential of displacing the presently used conventional bioassays.


Priming of interferon action.

Priming of interferon action. Pretreatment of L cell cultures with low levels of mouse interferon caused a more rapid development of and a greater degree of protection with subsequent exposure to higher levels of mouse interferon than that observed for control cultures. Such priming was a specific event which required cellular RNA synthesis. Priming by mouse fibroblast interferon, mouse immune interferon, and human leukocyte interferon was directly related to the plaque reduction titer of each of the interferons on L cells.


Inactivation of human interferon by body fluids. We have shown almost all samples of serum, saliva, cerebrospinal fluid, fecal extract, bile or urine tested contain a substance or substances capable of inactivating human interferon at normal body temperature. This adverse effect appears to be greater for fibroblast interferon-induced by either polynucleotides or viruses, than for leukocyte interferon induced by viruses. These results have a number of implications for the clinical utilization of this natural antiviral substance.


Activity of exogenous interferon in the human nasal mucosa. The activity of exogenous human leukocyte IF has been studied in the human nasal mucosa and factors have been sought that would enhance its potential for clinical usefulness. Experiments have been performed which indicate that the nasal epithelial cells do respond to exogenous IF. In vivo dose requirements may be high because of mucociliary clearance mechanisms and because of the time and dose requirement for full antiviral activity. Future studies must define the IF sensitivity of different respiratory viruses in the human nasal epithelial cell and must evaluate more efficient methods of application.


[Projections of the cortical visual areas of the posterior gyrus sigmoid and the caudate nucleus in the cat]. The complex performances of the visuo-motor system entail probably an intervention of circuits connecting, the primary visual areas, to other cortical regions, specially the sensory motor cortex, and certain sub-cortical formations. For testing this hypothesis, the unilateral resection of the areas 17, 18 and 19 has been undertaken on 19 cats, with delays ranging from 8 days to 3 months after intervention. The tracing of the pathways was carried out by combining different degenerating methods in particular the Marchi reaction. Based on this, it is possible to define a compact bundle of the axons originating from the primary visual cortex and dividing into two fascicles of unequal magnitude. The slender ends in the lower part of the posterior gyrus sigmoïd; the larger one penetrates into the caudale nucleus. An ultrastructural study specifies the modalities of distribution of the axons within this nucleus.


[Retinal ganglion cells of the albino rat: a qualitative and quantitative study]. Ganglion cells were investigated qualitatively and quantitatively in whole mount preparations of the albino rat retina, stained with cresylviolet. Cells of different size are distributed in different density over the retina, mainly arranged radially. The ganglion cell density varies between about 1200/mm2 at the edge of the retina and 5000/mm2 in the central area, centered about 1,5--2 mm away from the optic disc in the upper temporal quandrant. The mean soma diameter of the ganglion cells ranged from 7--23 micron. Based upon soma diameter histograms with regard to the eccentricities of the area centralis, ganglion cells were classified into three groups: small (7--10 micrometer diameter) mediumsized (11--14 micrometer) and large (15--23 micrometer). Relative frequencies of these groups were 53, 40 and 7% respectively. In the area centralis is the highest density of small cells. Here the relative frequencies were 62, 34 and 4% respectively. The diameter of the retina runs to 8 mm and the area to more than 40 mm2. In 2 retina the total number of ganglion cells were about 108000 and 111000 respectively.


A layered rhabdom in an isopod (Crustacea). A case of convergent development. The isopod species Astacilla longicornis (Sowerby) has a layered rhabdom, which is the first case reported in isopod crustaceans. The rhabdom comprises the rhabdomeres of six retinular cells. It is surrounded by an extracellular palisade. Usually, palisades are intracellular in arthropods. A layered rhabdom is found in taxonomically widely separated groups within the Arthropoda and no intermediate forms are found between the different, highly specialized rhabdom types present. It is concluded that the layered rhabdom represents an example of convergent development.


[Differentiated vascularization of the Dogiel cell types and the preferred vascularization of type I cells in the ganglia of plexus submucosus externus (Schabadasch) of the swine]. 1. In analyzing the variations in form and density of the periganglionic capillary networks of the plexus Schabadasch there could unequivocally be shown a manifold better vascularisation of type I cell aggregates, compared to those of type II. 2. The preferred vascularisation of type I cells is discussed in relation to the function of the cell types of Dogiel.


Histo-physiological studies on the interrenal gland of a catfish, Clarias batrachus (Linn.). The effects of three drugs namely, reserpine, atropine, and adrenaline have been investigated on the interrenal gland of Clarias batrachus. All the drugs bring about lesser or greater degree of hypertrophy in the interrenal gland of this fish. Atropine exhibits more vigorous effect than reserpine and adrenaline.


[Quantitative study of the apical dendritic spines in the cingulate lamina-V-pyramid neurons of the rat following oral application of alcohol]. The question answered in this paper is whether the apical dendritic spines-values of lamina-V-pyramidal cells of the gyrus cinguli of the rat are changed as a result of the oral application of ethanol. Alcohol-application causes a statistically significant increase of apical spines, especially at the main dendrite when given during the early postnatal development phase, where the brain is still impressible. The results are discussed as compensatory achievement of the not degeneratively changed neurons, while other pyramids show signs of degeneration. The conclusions were compared with the results of Kunz et al. (1976), who investigated the ethanol-effects on hippocampal pyramids.


Light and electron microscopy observations in the thyroid gland of the pheasant (Phasianus colchicus L.). In the summer, the pheasant thyroid presents as a relatively quiescent organ with reduced secretory activity; the follicular epithelium is very low and mostly of the pavement type, while the follicles are unwontedly large and contain an increased amount of colloid. The follicular cell cytoplasm contains numerous characteristic spherical vacuolar structures. In the winter, however, the thyroid displays remarkable reconstruction and raised activity of the follicular cells, which are much higher and columnar; the small follicles contain less colloid. Peculiar rod-like or fusiform structures, some measuring up to 6,5 microgram, were observed in the cytoplasm of the follicular cells of both hen and cock pheasants in November and March. At the beginning of the winter (November), specific dark cells with pycnotic nuclei, evidently in process of degeneration, appeared in the follicular epithelium. In March, specific granulated cells (probably parafollicular cells) were found sporadically in the immediate vicinity of the wall of subcapsular follicles in cock pheasant.


The male accessory reproductive glands and spermatophore in Hydrophilus olivaceus (Polyphaga-Coleoptera). 1 In Hydrophilus olivaceus two pairs of accessory glands of mesodermal origin are associated with the male reproductive apparatus. The inner pair of glands (AG1) are thin-walled, recurved tubes of uniform diameter. Their histology and histochemistry was described. They elaborate muco-or glyco-t proteins with no RNA component and some free glycogen. The mode of secretion is merocrine. The secretory products are used for the formation of sperm capsules packed inside the spermatophore and possibly for nourishing the spermatozoa. 2. The outer pair of massive, thick-walled accessory gland (AG2) is distinguishable into 4 zones of varied histological structure, which was described in detail. The powerful external muscular coat projects inwards in the form of septa that divide the secretory epithelial cells into compartments, each having its own lumen and all the lumina lead into a central lumen. The mode of discharge of secretory products involves both merocrine and holocrine methods. The secretory cells seem to undergo endopolyploid growth of their nuclei before participating in the synthetic activities. The secretory products of AG2 are muco- or glyco proteins that tend to be coupled with RNA. These are probably utilized for the formation of the massive eosinophilic ring and pigment granules that surround the helically coiled sperm column of the spermatophore. 3. Morphology and histology of the spermatophore, after its deposition in the female genital tract, was described in detail. 4. Possible mechanisms of spermatophore disintregration and its removal from the spermathecal barrel are discussed.


Hypothalamo-hypophysial neurosecretory system of the teleost Trichogaster fasciatus (Bloch & Schneider) with a note on it's vascularization. The hypothalamo-hypophysial neurosecretory (HN) complex of Trichogaster fasciatus is described with the help of in situ staining technique and tissue sections. The neurons of the nucleus preopticus (NPO) give rise to fine neurosecretory axons which form the left and right main tracts. The bulk of the tracts come laterally and enter the middle portion of the neurohypophysis. However, some of them spread out and penetrate the rostral neurohypophysis as well. Hypophysial artery contributes to the formation of primary capillary plexus (PCP) which extends from the subterminal region to the extremity of the anterior neurohypophysis. Structurally, the subterminal region and the anterior neurphoyophysis can be compared to the median eminence (ME) of tetrapods as it is differentiated into ependymal, fibrous and reticular layers. Also, these areas have aboundant neurosecretory and silver positive axons suggesting the possibility of direct transmission of neurohormones into the blood.


[Fluorescence histochemical studies of the uptake of exogenous noradrenaline and dopamine by in vitro cultured cerebrocortex explants of the rat]. Cerebrocortex of the neonatal rats were cultivated (--14 days). The cultures were studied living and with histological and fluorescence histochemical methods. A differentiation of neuronal cell- and fiber elements, oligodendro glial cells and astrocytes was found. The glyoxylic acid technique to estimate biogenic monoamines (Lindvall et al. 1974) was adapted up the cultivated explants. The normal cultures have only 24 h post cultivationem a specific fluorescence granularly in small concentration of the surface of the explant and in the explant self. Incubations with noradrenaline and dopamine demonstrated a various accumulation of the exogenous transmitters in the various parts of the cultivated explants. Uptake and releasing mechanisms in the cultivated material of the cerebrocortex were discussed with respect to the results of the sympathetic ganglia in vitro.


Pituitary gland of a hillstream sisorid catfish, Pseudecheneis sulcatus (McClelland). Pituitary gland of a hillstream sisorid catfish, Pseudecheneis sulcatus has been studied light microscopically. The pituitary is suspended from the hypothalamus by a slender infundibular stalk which enters the gland antero-dorsally, giving rise to the dorso-elptobasic condition. The pituitary is demarcated into pars nervosa, rostral pars distalis, proximal pars distalis and pars intermedia regions. Pars nervosa is formed of neurosecretory fibres which ramify extensively in the pars intermedia. The rostral pars distalis has predominantly acidophils which are arranged in rosettes. Proximal pars distalis has cyanophils, acidophils and few chromophobes while only acidophils and cyanophils are present in the pars intermedia. Sparse cyanophils and chromophobes are also present in the rostral pars distalis. The acidophils of proximal pars distalis are of two types which differ in tinctorial and morphological characteristics, from the acidophils of rostral pars distalis. The cyanophils are positively stained with AF, AT, PAS and aniline blue.


The nucleus of Darkschwitsch in Pantodactylus schreiberii Wiegmann (Lacertilia, Reptilia). The nuclear mass considered the representative of the nucleus of Darkschwitsch in reptiles in studied. It is composed of small, stellate, pyriform or rounded cells, with two or three dendritic trunks resolved in a plexus near the cellular body. Its axons take an ascending direction to the posterior commissure, and not to be medial longitudinal fasciculus as has been conventionally described.


The behaviour of pigment cells in the leg muscle compartments of Silver Campine fowl. The presence of pigment cells in the leg muscle of Silver Campine fowl is ascertained. Transverse sections of legs of 16 and 21 day embryos gave similar results. Analysis of the states of differentiation indicates three types of pigment cells contained by the leg muscles. The relationship between the tissue environment and the melanoblast differentiation is discussed.


SGC (SIF) cells in autonomic ganglion: evidence for a possible secretion of their content into the blood vessels. Authors described signs of extrusion of granular vesicles from the SCG (SIF) cells in the inferior mesenteric ganglion of the dog. The granular vesicles were found in extracellular space as well as in the blood capillary lumen. Beside this, large number of granular vesicles were found in the cytoplasm of endothelial cells of blood capillary, mainly in places of close contact between the SGC cell and the endothelial cell of the capillary. These findings strongly suggest secretory function be a primary role of SGC cells in inferior mesenteric ganglion and would explain the rich vascularisation of the SGC cell groups.


Effect of neurectomy on nuclease activity in skeletal muscles of rats. Author followed up the activity of the three enzymes involved in the catabolism of nucleic acids--acid deoxyribonulease (DNase II), alkaline ribonuclease (RNase I), and acid ribonuclease (RNase II)--in the denervated gastrocnemius and soleus muscles of rats for 28 postoperative days. The activity of both acid nucleases increased in both types of denervated muscles, compared with the respective controls. Up to the 14th postoperative day, the activity excess of both acid nucleases was more significant in the m. gastrocnemius than in the m. soleus. The RNase I ran below the control activity during the whole period in the m. soleus and up to the 14th day in the m. gastrocnemius. The role of nucleases and nuclease inhibitors in the changes of nucleic acid catabolism in neurogenic muscular atrophies is discussed.


Cellular autophagocytosis in mouse seminal vesicle cells in vitro. Cellular autophagocytosis was observed in mouse seminal vesicle cells incubated in vitro up to 8 h in medium 199 or Krebs-Ringer bicarbonate buffer. During the first 2 h of incubation, early forms of autophagic vacuoles were seen in the cells, advanced forms containing degraded material began to cumulate later. After 6--8 h, early vacuoles occurred sparsely, while advanced forms were detected in a great number. During the first 2 h of incubation, we often observed smooth surfaced membrane pairs between the cisternae of rough surfaced endoplasmic reticulum resembling isolating membranes of autophagic vacuoles. They varied in size and shape from short, straight cisternae to long, curved ones, almost completely encircling areas of the cytoplasm. Based on these observations, we propose a tentative scheme of the formation of autophagic vacuoles, viz., the short, straight cisternae would represent the first stage in the development of an autophagic vacuole, while the curved sack-like forms are interpreted as successive steps leading to the complete sequestration of an area of the cytoplasm.


Quantitative characterization of the autophagic process in mouse seminal vesicle cells in vitro. Pieces of mouse seminal vesicles were incubated in vitro for 0.5, 1, 2, 4, 6, or 8 h and the autophagic vacuoles formed in the epithelial cells during incubation were counted. The vacuoles were defined as early or advanced forms by morphological criteria and the autophagic vacuole/cell ratios were determined for the early and the advanced autophagic vacuoles separately. The number of early autophagic vacuoles significantly increased in the first hours of incubation, it was the greatest in the 4th h and rapidly declined thereafter. The number of advanced vacuoles began to rise about 1.5 h later as compared to the early ones, reached its maximum in the 6th h and remained high even in the 8th h. It may be concluded that a wave-like process took place in these cells during the experiment, characterized by a short transient rise in the formation of autophagic vacuoles, and their subsequent transformation into advanced forms filled with degrading cytoplasmic fragments.


Identification and estimation of an acetylcholine-like substance in the venom of Dendroaspis jamesoni. An acetylcholine-like substance was isolated from desiccated Jameson's mamba venom by one-dimensional ascending paper chromatography. The migratory and staining properties of the substance were identical with those of standard acetylcholine. Pharmacological identification on various in vitro and in vivo biological test objects showed that the substance was acetylcholine. It was further confirmed by high-voltage paper electrophoresis. The acetylcholine content was 4.35--4.36 mg/g of desiccated venom as shown by two different biological assay methods, and the index of discrimination was found to be 1.1, further confirming that substance in snake venom was acetylcholine.


Electron microscopic investigation of a giant neuron identified in the right parietal ganglion of Lymnaea stagnalis (L). The ultrastructure of a giant neuron (GRP1) identified in the right parietal ganglion of Lymnaea stagnalis was examined. The results suggest that GRP1 is a neurosecretory cell. The perikaryon is characterized by many neurosecretory-like granules of variable electron density. The average diameter (longer axis) of the granules was 1850 A. Furthermore highly-developed rough endoplasmic reticulum and Golgi complex were observed in the cytoplasm. Synapse-like structures were often found on the somatic membrane, but typical, true synaptic endings were not seen.


Microvascular procedures in the management of lymphedema. The burgeoning field of lymphatic research, and the applications of surgical and microsurgical techniques to the problem of lymphedema have been briefly reviewed. The chronicity of this symptom complex and the relative sparsity of cases in developed countries make controlled clinical trials of treatment unfeasable and comparisons of present regimens difficult. However, continued investigation will undoubtedly be fruitful.


Studies on bovine spleen cathepsin D.

Studies on bovine spleen cathepsin D. The purification procedure of cathepsin D which includes autolysis results in the destruction of the molecule to smaller polypeptide chains. Pure catepsin D obtained by the method which includes affinity chromatography, contains single polypeptide chain of 42000 daltons. The N-terminal amino acid is glycine. The specificity was studied using synthetic substrates. CD measurement of cathepsin D shows mainly unordered structure, about 26% of beta-structure and only 5% of alpha-helix. Binding of pepstatin shows pronounced changes in the CD spectrum between 250 and 300 nm; above 7.5 no interaction was observed.


The effect of human neutrophil elastase and cathepsin G on the collagen of cartilage, tendon, and cornea. The degradation of collagen in a number of inflammatory diseases is associated with infiltration of the tissues by neutrophil leucocytes. The neutrophil serine proteinases, elastase and cathepsin G, solubilize type II collagen from articular cartilage by an attack on the terminal peptides, destroying the inter- and intra-molecular crosslinks. Both enzymes also solubilize type IV collagen from kidney basement membrane. Elastase attacks the terminal peptides of type I collagen of tendon, cornea and skin, and also attacks the helical region of type I skin collagen. It seems probable that the neutrophil proteinases play an important role in collagen degradation in vivo in a variety of pathological conditions.


Protein cleavage in virus-infected cells. A variety of proteins, including viral precursor polypeptides, were bound to a solid support and used in a sensitive assay for proteolytic enzymes in HeLa cells. A trypsin-like endoprotease, present on ribosomes of HeLa cells, loses activity after picornavirus infection. The decline follows synthesis and processing of a viral protein. Inhibition of cellfree activity of HeLa protease occurs when protein trypsin inhibitors or double-stranded RNA are added. After the mid-point of infection, protease activity with enhanced specificity for viral substrates is detected. The new protease has a pH optimum and heat stability different from endogenous host enzymes, and is synthesized following infection. A viral mutant was isolated which produces a temperature-sensitive protease. The results indicate that a poliovirus gene product participates enzymatically in the final cleavages of some polioviral proteins. A model for the regulation of poliovirus replication based on specific proteolysis is presented.


Peptidases of the kidney microvillus membrane. The microvillus membrane of the kidney is highly differentiated in its complement of enzymes and other proteins. In addition to the five well documented peptidases that are present in the membrane, recent work suggests that aminopeptidase P, carboxypeptidase P and an enzyme tentatively referred to as "leucine hydrazidase" are also microvillus enzymes. Microvillus serine peptidase (dipeptidyl peptidase IV) has been purified after detergent solubilization. The catalytic and molecular properties of this form and the form released by autolysis have been compared in an attempt to gain understanding of the intramembranous domain of this protein. Current views on the molecular organization of the microvillus are discussed.


Degradation of myofibrillar proteins by cathepsins B and D. Cathepsin B and cathepsin D were purified from rat liver and skeletal muscle. Electrophoretic analyses revealed that the enzymes were highly purified, and isoelectric focusing demonstrated multiple forms of both enzymes. Purified actin and myosin, as well as actin and myosin in myofilaments and myofibrils, were degraded by the purified cathepsins B and D. Degradation of myosin was completely blocked by the cathepsin B and D inhibitors, leupeptin and pepstatin, respectively. Cathepsins B and D were visualized by electron microscopy, using CBZ-Ala- Arg-Arg-4-methoxy-beta-naphthylamine and BZ-Arg-Gly-Phe-Leu-4-methoxy-beta-naphthylamine as substrates.


Degradation of myones as a consequence of disuse and denervation. After denervation or immobilization in both slow and fast muscles of rabbits a rapid decrease in weight and protein content and a focal degeneration of cell ultrastructure were observed. As a consequence marked changes in the ratio of soluble to myofibrillar protein and in regulatory proteins of myofibrillar system occurred.


A neutral protease from rat intestinal muscle. A possible role in the degradation of native enzymes. A membrane-limited protease has been solubilised and partially purified from the intestinal smooth muscle of rats fed on protein free diets. This neutral protease has a mol. wt. of around 33,000 and from its susceptibility to several known modifiers of proteolytic enzymes, it appears to be trypsin-like. It is stable over a relatively narrow pH range and it appears to have a markedly enhanced ability over trypsin for inactivating substrate enzymes in their native conformations through limited proteolysis. The rate of inactivation of substrate enzymes can be modulated by cofactors, allosteric ligands, or by changes in ionic strength. In addition, a specific protein inhibitor of the protease has been measured and levels of this are high in animals fed on normal diets. On administration of protein free diets, the inhibitory activity is depleted. Contamination of the muscle tissue by lumenal, mucosal or blood proteases and inhibitors has been excluded. A role for the neutral protease in initiating the turnover of intracellular enzymes is postulated.


The susceptibility of glycogen phosphorylase to inactivation by endogenous and exogenous proteases. Phosphorylases a and b were inactivated very rapidly by a neutral, trypsin-like protease from rat intestinal muscle. With 32P-phosphorylase a as substrate, it was shown that the initial event in the inactivation was the release of a small, phosphopeptide from the N-terminus of the enzyme, leaving the original 100,000 subunit form virtually unchanged. Subsequent proteolysis was very limited, producing 85, 70 and 65,000 mol. wt. derivatives. The effects of several allosteric modulators of phosphorylase on the rates of inactivation of the two enzymes were studied. Removal of the pyridoxal phosphate cofactor from phosphorylase increased the susceptibility of the b form by three fold while the a form was unaffected. By comparison of these effects with those obtained from digestion with trypsin and chymotrypsin, it is concluded that the intestinal muscle protease has a markedly enhanced ability for inactivating enzymes in their native conformation. Assuming that this property is reflected in vivo, a possible role such neutral proteases in initiating protein degradation is advanced.


Evidence pointing to the main role of lysosomes in mitochondrial proteolysis at neutral pH. Recombination experiments using radioactive mitochondria and mitoplasts, and nonradioactive lysosomes or digitonin-soluble fraction of mitochondria, show equal rates of proteolysis and of inactivation of carbamyl phosphate synthetase; the amount of lysosomal protein was equal in both cases on the basis of N-acetyl-beta-glucosaminidase activity. Therefore, lysosomes seem to be responsible for all the proteolytic activity exhibited by the digitonin soluble fraction of mitochondrial preparations. Since this fraction contains ca. 90% of the proteolytic activity present in mitochondrial preparations, most of the proteolysis can be attributed to lysosomal contamination. These findings and stability characteristics "in vitro" and "in vivo" of some matrix enzymes are presented and discussed in relation to protein turnover.


Increased susceptibility of carbamylated glutamate dehydrogenase to proteolysis. Glutamate dehydrogenase is very susceptible to carbamylation which results in loss of activity. The effect of a number of proteolytic enzymes (pronase, trypsin and chymotrypsin) on native and carbamylated glutamate dehydrogenase was tested. In all cases, the carbamylated enzyme was at least twice as susceptible to proteolysis as the native enzyme. Antibodies were prepared against glutamate dehydrogenase and carbamylated glutamate dehydrogenase; the carbamylated enzyme was antigenically indistinguishable from the native enzyme. Preliminary experiments indicate that the carbamylated glutamate dehydrogenase is taken up by ascites tumor cells while glutamate dehydrogenase is not. It seems possible that the effects described can be extrapolated to degradation by lysosomes and to other covalently modified enzymes.


Acetyl glutamate--a model of signals for intracellular proteolysis. The proteolysis at neutral pH of mitochondria from liver and brain is more marked in isolated preparations than "in vivo" indicating activation of proteases or inactivation of repressors during isolation. Acetyl glutamate (AG), found in liver mitochondria of ureotelic animals, plays a crucial role as activator of carbamylphosphate synthetase. Since AG levels change under a number of conditions, we checked for an AG deacylase in mitochondria, for otherwise AG must be exported and destroyed by cytosol deacylases. We noted on incubation of mitochondrial extracts with AG an increase in trichloracetic acid-soluble ninhydrin-reacting material but not in acetate liberation, indicating activation of proteases. This was checked with 14C-labelled mitochondria. Under certain conditions AG and other acyl aminoacids stimulate approximately 5 to 20% the proteolysis with rat live and with brain mitochondria.


Flow and shuttle of plasma membrane during endocytosis. A striking feature of endocytosis is the large amount of surface membrane that is brought into the cells through the formation of endocytic vesicles. Little is known about the fate of this membrane material. It is implausible that it would be destroyed in lysosomes, as the rate of turnover of the constituents of plasma membrane is much too low with respect to the rate of endocytosis in all cells studied so far. Conversely, plasma membrane fragments, internalized by endocytosis cannot merely be incorporated in lysosomes, as these organelles have been shown to maintain their size, despite continuous and active endocytosis. We present evidence that plasma membrane antigens, detected by means of specific antibodies, are internalized during endocytosis and reach lysosomes. They are thereafter returned back to cell surface. These results indicate the existence of a shuttle of membrane elements between the cell surface and lysosomes.


Inhibition by insulin of the physiological autophagic breakdown of cell organelles. By using electron microscopic morphometry it was found that the fractional volume of autophagic vacuoles in the cytoplasm of liver cells decreased rapidly after administering 5 units insulin per kg body weight to otherwise untreated rats. From the decay which appears to follow first order kinetics a half-life of 9 min was found for the compartment of autophagic vacuoles. The rates of autophagic breakdown calculated were different for the different cytoplasmic components, indicating that selecting mechanisms are involved in the process of segregation which is the first step in autophagy.


Studies on the relationship between the molecular structure and the catabolism of insulin. The catabolism of insulins modified at the A1, B1 or B29 positions or containing a synthetic crosslink between the A1 and B29 positions has been studied in vivo and in vitro. The metabolic clearance rates (MCR) of insulin, proinsulin and chemically modified insulins have been measured by a priming-dose constant infusion technique in greyhounds. Insulins modified at A1 and B29, particularly the crosslinked materials, had markedly lowered MCR's whilst B1 analogues did not differ from insulin. Proinsulin and the A1-B29 crosslinked materials showed a markedly lowered degradability by glutathione-insulin transhydrogenase.


Conversion of proinsulin into insulin by cathepsins B and L from rat liver lysosomes. Conversion of proinsulin and intermediate forms of proinsulin into insulin were studied with rat liver cell fractions and purified lysosomal proteinases by using the technique of polyacrylamide disc-electrophoresis. Both substrates were degraded very rapidly by homogenates and crude lysosomal fractions to split products not detectable on disc-electropherograms. Neither breakdown nor conversion were detected with the cytosol and the microsomal fraction. With partially purified lysosomal fractions (mol. wt. approx. 25 000) or with highly purified cathepsin L or cathepsin B (B1) proinsulin was converted into products migrating like the intermediate forms and insulin, and the intermediates were converted into products migrating like insulin and deoctapeptide-insulin in disc-electropherograms. The mechanism of conversion seems to be different for both enzymes. The results force us to conclude that lysosomal cathepsins, especially cathepsins L and B might be involved in the process of conversion of proinsulin into insulin and perhaps also of other precursors into biologically active proteins in vivo.


Presence of an endopeptidase activity in rat liver ribosomes. Preparation of ribosomes using different procedures (treatment of postmitochondrial-postlysosomal supernatant or microsomes with 1% triton in 0.15 or 0.5 M KCl and subsequent sucrose gradient centrifugation; treatment of microsomes with 1.5% deoxycholate/2% triton) results in purified ribosomes which contain an endopeptidase activity detectable by breakdown of ribosomal proteins to trichloroacetic acid soluble split products. The proteolytic activity can be recovered also in the extracted proteins of whole ribosomes. With ribosomes the pH optimum of proteolytic breakdown is at about 7. The inhibition of the activity by leupeptin, DIFP and soya bean trypsin inhibitor suggests a serine type of the proteolytic activity.


The role of lysosomal enzymes in protein degradation in different types of rat liver cells. Highly purified suspensions of parenchymal, endothelial and Kupffer cells were prepared from the rat liver. The respective roles of these cell classes in the degradation of proteins was investigated by analysing the cellular distribution of two lysomal proteases. The specific arginine naphthylamidase activity was 2 times higher in Kupffer cells compared with the nearly equal activities in endothelial and parenchymal cells. The specific activity of the important endopeptidase cathepsin D in endothelial and Kupffer cells was about 12 and 36 times higher, respectively, than the activity in parenchymal cells. These results are in agreement with an important role of Kupffer and endothelial cells in the degradation of proteins and protein containing material of exogenous origin.


Endocytosis and breakdown of proteins by sinusoidal liver cells. We studied the uptake of modified forms of bovine pancreatic ribonuclease A (labeled with 125-iodine) by rat liver in vivo. On one hand, these experiments were intended to investigate a possible role of sinusoidal cells in the uptake of plasma proteins; on the other hand, the effect of well-defined modifications of the enzyme on the role of uptake might give us a key to the factors determining life time of plasma proteins. We used nephrectomized rats in most experiments to avoid uptake by the kidneys. Preparations of ribonuclease oligomers prepared by cross-linking with dimethyl-suberimidate enabled us to study a possible relation between uptake and molecular size. This modification does not lead to changes in charge of the protein. Monomer, dimer and polymer fractions were isolated by gel filtration on Sephadex G-75. Of the 11 amino groups in ribonuclease A, 9, 8 and 6 remained unaltered in the monomer, dimer and polymer fraction, respectively. The maintenance of biological activity, the stability of disulfide bonds, and the unchanged susceptibility to endoproteases of the cross-linked products established that gross conformational changes had not occurred. At 1 h after injection, 1% of themonomer, 7% of the dimer and 19% of the polymer were recovered per g of liver protein. Combination of autoradiography, subcellular fractionation, and the determination of labeled ribonuclease derivatives in the spleens showed that the dimer and polymer fractions were mainly present in the lysosomes of sinusoidal cells.


Attempts to relate enzyme inactivation to degradation in vivo. Inactivation of liver cytosol proteins has been measured in vitro in the presence of various membranes and disulphides. Inactivation rates correlate with the known degradation rate constants of the enzymes in the intact liver. More extensive studies were carried out with glucose-6-phosphate dehydrogenase (G6PD) and phosphoenolpyruvate carboxykinase (PEPCK) using either cytosol as a source of these enzymes or alternatively highly purified preparations of each enzyme. All membranes purified from liver had a considerable capacity to inactivate the enzymes with higher activity found in the hepatocyte plasma membrane. Various lipid preparations or plasma membranes from other tissues were virtually ineffective. Inactivation was dependent on disulphides in the membranes as shown by the inhibition of activity if membranes were pretreated with thiols. Preliminary experiments of the fate of inactivated G6PD or PEPCK show binding to membranes and subsequent proteolysis. A model is proposed for the degradation of labile enzymes.


Lysosomes and protein degradation.

Lysosomes and protein degradation. Evidence from studies on mouse peritoneal macrophages using the inhibitor pepstatin confirms lysosomal involvement in basal protein degradation, and extends its relevance to degradation of long half-life and analogue containing proteins. Studies on the ability of MRC-5 (a limited life-span fibroblast line) cells to selectively degrade analogue-containing proteins are described. These indicate that this capacity is retained even in very old cells; indeed such cells show an increased proportion of rapidly-degradable proteins. Analogue containing proteins bind preferentially to lysosomal membranes, and like liver cytosol proteins of short half-life, are selectively endocytosed and degraded by certain cells in culture. Thus membrane binding allowing selective entry to the lysosomal system may be important in controlling rate of degradation of both intracellular and extracellular protein. A method potentially allowing for determination of the rate of autophagy in cells, is described. This should enable further assessment of the quantitative involvement of lysosomes in protein degradation.


Protein degradation in rat liver cells.

Protein degradation in rat liver cells. The intracellular protein catabolism in isolated hepatocytes is inhibited by pepstatin and particularly by leupeptin in the presence of liposomes. Leupeptin has an inhibitory action both in the absence and in the presence of liposomes (nearly 40%).


Pepstatin- and leupeptin-loaded liposomes: a tool in protein breakdown studies. The preparation of proteinase inhibitor loaded "solid" positively charged liposomes is described. Some properties of the vesicle preparations are given and the findings are tentatively summarized in a scheme of liposomophagy.


Cooperation of various subcellular fractions in protein degradation in vitro. In order to test the possibilities in protein degradation between cell organelles comparatively, [3H]- and [14C]-leucine short-time labelled subcellular fractions from rat liver were incubated with each other at pH 6.9. All fractions tested were able to degrade short-lived proteins from foreign fractions, whereby the lysosomal supernatant fraction showed the highest proteolytic activity, which declines in the sequence: lysosomes--nuclei--mitochondria--cytosol--microsomes. Short-lived cytosolic proteins were especially suited as substrate for neutral proteases from all other fractions, but also microsomal, mitochondrial and nuclear proteins were well degraded by foreign fractions in comparison with the substrate autoproteolysis. Therefore in vivo manyfold cooperations between several organelles in protein catabolism seem to be possible.


Aging changes in intracellular protein breakdown. Using radioactively labelled cytosol proteins as substrates we were able to exclude the possible accumulation of any specific inhibitor for the lysosomal proteases in rat liver cytosol during the aging process. There were also no gross changes in the molecular weight patterns of these proteins during the aging process. The percentage of more hydrophobic proteins seems to be identical in both the "old" and "young" cytosol proteins. From immunological experiments we suppose a qualitative change in the composition of rat liver cytosol proteins or of their properties during the aging process.


Naturally occurring inhibitors of intracellular proteinases. The papain inhibitor isolated from chicken egg white inhibits the enzymatic activity of cathepsin B1 and cathepsin C. The inhibitor bears two nonoverlapping reactive sites: one binds cathepsin B1, papain, ficin, and bromelain, the other one cathepsin C. The inhibitor decreases the degree of an immunologic hypersensitive reaction, the so-called Arthus reaction. A statistically significant inhibition of this immunologically developed inflammation occurs only if the inhibitor is applied intradermally and simultaneously with the provoking dose of the antigen to rabbits sensitized to the same antigen. The pepsin inhibitor from the body walls of the roundworm Ascaris lumbricoides inhibits the proteolytic activity of cathepsin E. This inhibitor covalently bound to Sepharose 4B was used for affinity chromatography of cathepsin E. A cathepsin D inhibitor was isolated from potato tubers and its inhibitory and chemical characteristics were studied. The inhibitor does not inhibit either cathepsin E or pepsin yet inhibits trypsin in the alkaline pH-range. The molecular weight of the inhibitor is 21 790 and its molecule consists of 199 amino acid residues. The sequence of 17 amino acid residues was determined by Edman degradation of the inhibitor molecule.


Isolation and characterization of inhibitors of neutral proteinases from spleen. Two different types of neutral proteinase inhibitors were isolated from postmicrosomal supernatant of bovine spleen. Inhibitor A with molecular weight 40,000 inhibits elastases and chymotrypsin-like neutral proteinases from bovine spleen, whereas inhibitor B with estimated molecular weight 20 000 inhibits only chymotrypsin-like neutral proteinase.


Studies on the in vivo-action of leupeptin on the nitrogen retention in rats. A daily s.c. injection of 0.5 mg leupeptin for 7 days does not influence the digestibility of food-N but seems to improve the N-retention. A daily injection of 1.0 mg leupeptin for 14 days is without influence on the digestibility of N, too, but the N-retention was decreased. At this leupeptin level the half-lives of intestinal proteins were prolonged. The data indicate the possibility to influence the N-retention using leupeptin.


Inactivation studies of cathepsin D with diazo compounds. Cathepsin D was inactivated with various diazo compounds at very high concentration. Reaction proceeded maximally at pH 4.5 in the presence of cupric ions. With 3-diazo-indazole and triazene the inactivation was noted also in the absence of cupric ions what indicates that the mechanism is mediated through triazene. CD-spectrum of partially inactivated enzyme shows that conformational changes occurred after treatment with diazo compound.


Inhibition of serine proteinases by benzamidine derivatives. Derivatives of benzamidine inhibit competitively the activity of the serine proteinases trypsin, plasmin, thrombin, and of the clotting factor Xa. The inhibitor activities (Ki-values) of various benzamidine derivatives against the several enzymes were compared. Besides parallels, deviations in the corresponding structure-activity relationships were found. From these results it is concluded that the similar enzymes exhibit certain differences in the structure of the primary and secondary binding sites.


Endogenous proteolytic activity of chromatin. Chromatin from rat liver and Ehrlich ascites tumor (EAT) was isolated by two different procedures and the chromatin preparations were incubated at 37 degrees C. To follow the proteolysis at 0, 4, 8, and 24 hrs, aliquots were taken and analysed by SDS polyacrylamide gel electrophoresis. Although there were some differences in the proteolytic activity of the different chromatins, in general they were found to sustain a several hours incubation without appreciable degradation to occur. However, when prior to incubation the chromatins were dissociated, a rapid proteolysis took place. This shows that there are specific, chromatin bound proteinases that are inactive while immobilized in the chromatin structure and become activated on dissociation.


Degradation of phosphorylated chromosomal nonhistone proteins. In experiments with in vivo 32P-labelled nonhistone proteins of rat liver nuclei it was shown that these components are more sensitive against degradation than the mass of the nonhistone proteins. In the presence of 0.1 mM phenylmethylsulfonylfluoride and 1 mM sodium molybdate, however, they are protected against degradation.


Interaction of intracellular proteases and immune mechanisms. Intracellular proteases play an important role both in nonspecific and specific immune reactions. Results concerning the interaction of these enzymes with phagocytic factors responsible for killing microorganisms are presented. In inflammatory foci, proteases released from destroyed leukocytes have been found to modify the function of antibodies present. They particularly reduce their complement fixation activity. Macrophages with differing proteolytic activity demonstrated diverse effects on the formation of antibodies tested by Jerne's plaque technique in young rabbits immunized with sheep erythrocytes. Whereas alveolar macrophages diminished antigen immunogenicity, peritoneal macrophages enhanced it.


The diversity of cellular proteinases in physiology and pathology. 1. It is suggested that the functions of cellular proteinases are far more diverse than has usually been realized. 2. A scheme is proposed in which isolation of a pure proteinase leads to a rather detailed exploration of its biological significance. 3. Representative proteinases from the serine, thiol, carboxyl and metallo-classes are briefly reviewed with emphasis on the relationship between improved understanding at the molecular level and eventual progress in the understanding of some important physiological and pathological processes. 4. An attempt is made to forsee future developments in this area of biochemistry, in terms of both new trends in research, and the eventual application of knowledge of the proteinases to medical problems.


Type B hepatitis in Iran.

Type B hepatitis in Iran. Hepatitis B surface antigen (HBsAg) was found in 1% of controls, 2.1% of professional blood donors, 2.0% of leprosy patients and 76.1% of acute hepatitis in Tehran and Mashhad, Iran. All HBsAg positive samples also possessed antibody to the hepatitis B core antigen and all were subtype ayw. Type B hepatitis and the HBsAg state are frequent in Iran, but most must be accounted for by "non-parenteral" or inapparent" parenteral exposure.


The concept of orthodontic treatment for the adult. Through our experience gained with adult patients during the last ten years we have found it useful to divide them into three groups:- 1--Those who are very strongly motivated to accept orthodontic treatment for cosmetic purposes and with whom treatment will usually be satisfactory. 2--Those who are suffering from anxiety states and who have developed a fixation on a dental fault and with whom it is very unwise to begin treatment before a favourable psychiatric report is obtained. 3--Those patients who need treatment before prosthetic rehabilitation or for other physical disorders and in whom treatment may or may not be successful, depending on the degree of co-operation shown by the patient and on good teamwork between dental specialists.


A new hemoglobin beta chain variant in sheep. A new hemoglobin beta chain variant referred to as Hb-E is observed in four Indian sheep breeds. Hemoglobin E is electrophoretically and chromatographically undistinguishable from hemoglobin B. However, polypeptide chain separation by urea-polyacrylamide disc gel electrophoresis and urea CM-cellulose chromatography reveals the new beta chain. The betaE chain has a slower anodic mobility than betaB chain at alkaline pH. The adult sheep hemoglobins could be classified into eight categories, namely AA, AB, AE, AC, ABC, BB, BE and EE, according to their sub-unit composition. The occurrence of betaE chain in homozygous condition or in combination with either one of the two allelic beta chains betaA or betaB, indicate that it may be an allelic variant. Gene frequencies of three hemoglobin beta chain alleles observed during the study are as follows: A = 0.192; B= 0.682; E = 0.126.


Genetic variants of glucose phosphate isomerase (E.C. 5.3.1.9) in canine erythrocytes. Genetic polymorphism of glucose phosphate isomerase (GPI) was found in the erythrocytes of dogs of six Japanese breeds by using starch gel electrophoresis. Analysis of parentage records of dogs revealed that the phenotypic variation of erythrocyte glucose phosphate isomerase was controlled by one autosomal locus with two codominant alleles, GPIA and GPIB. The allele GPIB was observed in the following breeds: San'in-Shiba, Shinshu-Shiba, Shikoku, Kai and Kishu, but not in Hokkaidoes and Akitas. All the dogs belonging to 25 European breeds, 5 oriental or China-origin (except Japan) breeds examined in this experiments had the genotype constitution GPIA/GPIA, whereas one Dalmation dog was heterozygous GPIA/GPIB.


Serologically detected lymphocyte antigens in Holstein cattle. Lymphocytes from 103 Holstein cattle were tested with 11 antilymphocytotoxic sera. Four of these sera were produced by whole blood immunization; these generally yielded lymphocytotoxicity against a large number of animals in each test panel. The majority of the sera were collected from parous cows which had never been immunized. Observations about this latter group of sera are (1) lymphocyte antibodies, may occur by alloimmunization in foetal-maternal interaction, (2) normal sera from non-immunized parous cows generally had a narrower specificity of antibodies than from parous cows immunized with blood from unrelated animals and in some cases these antisera may be monospecific, and (3) these sera from parous cows are easy to obtain without the need for laborious immunizations. Analysis of these lymphocytotoxic sera indicate several reagents were obtained which possessed different antibodies; these were useful in detecting polymorphism of cattle lymphocyte antigens. Using these 11 sera, an analysis of the reactions patterns among 103 Holsteins, including parent-offspring data resulted in the postulation of several alleles. These data suggest that cattle lymphocyte antigens are very polymorphic and inherited.


Serum antigens of cattle. VI. Isolation and properties of an intracellular antigen (Ecl) from cattle erythrocytes. The erythrocyte antigen (Ecl) of cattle has been isolated using DEAE-cellulose chromatography, Sephadex gel filtration, and isoelectrofocusing. The purified antigen is free from hemoglobin and migrates as a single protein on polyacrylamide gels. It exhibits an absorption maximum at 278 nm and has a molecular weight of about 100 000; its isoelectric point is 4.95. The antigen has a carbohydrate moeity and loses its activity after oxidation with periodate. The evidence suggests that it is not a common isozyme, but is exact identity remains unknown.


Freeze drying cattle blood typing reagents. A system is described for freeze drying and storage of reagents (antisera) used in cattle blood typing tests. Reagents were freeze-dried in evacuated bottles at the desired dilution for rapid reconstitution to use in test procedures. Reagents not in current use were dried in bulk lots and stored in polyethylene film bags. All freeze drying procedures were performed with readily available commercial equipment. A computer program was developed to produce a current inventory of reagent supplies and projections of the number of samples which can be typed.


Serum amylase 2 polymorphism in pigs.

Serum amylase 2 polymorphism in pigs. A genetic polymorphism of the pig amylase 2 system is described. It is controlled by two codominant genes, Am 2A and Am 2B. A comparison of some breeds from Byelorussian breeding farms gives the following frequencies for the alleles Am 2A and Am 2B, respectively: 0.35 and 0.865 in Large White (n = 682), 0.257 and 0.743 in Byelorussian Black and White (n = 400), 0.540 and 0.460 in Hampshire (n = 51). While studying the genotype distribution according to the Hardy-Weinberg law, a genetic imbalance in Byelorussian Black and White pigs was established (chi2 = 56.4). Genetic relationship between the Am 1 and Am 2 loci was not found.


Radiation therapy of ureteral metastases from breast cancer. A series of 6 patients with ureteral metastases from breast carcinoma treated by radiation therapy has been evaluated. Most patients received a tumor dose of 3,600 rads in 3 to 4 weeks, and showed remarkable clinical improvement, with relief of the ureteral obstruction and reversal of symptoms and signs of uremia after radiation therapy. One patient died before an adequate dose of radiation therapy could be given. Early recognition and early treatment of ureteral metastases have been stressed. Radiation therapy techniques have been described.