Accumulation of histidine, 3-methylhistidine, and homocarnosine in the brains of protein-calorie deficient monkeys

Abstract— Seventeen monkeys (M. nemestrina and M. fascicularis) aged 10 months to about 5 yr were divided into two groups and fed either an adequate protein diet (20% casein) or a low-protein diet (2% casein). The diets were supplied to the animals in restricted amount (200 g/animal in two daily rations). In one experiment, the malnourished animals were initially fed a diet containiing 8 per cent protein and the protein content of the diet was gradually reduced over a period of 9 months, to 2 per cent. After about 3 months on the 2 per cent protein diet, the malnourished monkeys showed growth failure, severe anorexia, peri-ocular oedema, tremors of the head and limbs, atrophy of several visceral organs, fatty liver, hypoalbuminaemia, and depressed serum levels of many essential amino acids with an elevation of the ratio of non-essential to essential amino acids. These features are consistent with protein-calorie malnutrition. Examination of the brains revealed significant alterations in the levels of glycerophosphoethanolamine (—40 per cent), glutamic acid (—25 per cent), histidine (+230 per cent), homocamosine (+185 per cent), 3-methyl-histidine (+147 per cent), lysine (+55 per cent), phenylalanine (+33 per cent) and tyrosine (+26 per cent) in comparison to findings on the well-fed monkeys. The possible implications of elevated cerebral contents of homocarnosine in malnourished monkeys are discussed in the light of several reported human cases in whom neurological disorders are associated with increased histidine-containing dipeptides in the brain, CSF, blood and urine.     

Viremia and serum antibodies in Macacus rhesus monkey after inapparent infection with European tick-borne encephalitis virus.

1. Inapparent infection was called forth in M. rhesus monkeys by means of subcutaneous inoculation of ETBE virus. 2. Viremia was found in 18 (86 per cent) of 21 monkeys. In all the 18 monkeys, specific virus-neutralizing antibodies were found; in 17 of them complement-fixing antibodies developed in addition. 3. Neutralizing antibodies (N greater than or equal to 1.7 log10) were first recorded on 10th day with a peak on 32nd day, retaining, after a small decrease, a relatively high level in all reacting animals. 4. Complement-fixing antibodies were first found on 18th day, showing a peak on 22nd day whereafter they dropped considerably or even disappeared so that on 150th day they were present in a low titre in only 35 per cent of the originally reacting monkeys. 5. The findings were compared with the situation in naturally infected humans and in hamsters infected inapparently (experimentally) with ETBE virus.     

Malaria, arbovirus and hepatitis infections in Macaca fascicularis from Malaysia

Naturally occurring malaria, arbovirus infection and hepatitis in monkeys can be a hazard for the investigator and might interfere with the outcome of experiments. 63 young adult Macaca fascicularis from Malaysia were screened for these infections. About 1 year after their arrival in France, parasitaemia due to Plasmodium spp., was present in 6.4% of the animals and specific antibodies in 55.5%. 19 of 35 initially positive monkeys were tested again 2 years later. Parasitaemia was found in 1 of 4 monkeys and antibodies in 11 of 19 monkeys which were initially positive. 9 of the monkeys initially tested had low titres of antibodies to the Flavivirus genus. All animals were negative for the hepatitis B surface antigen and anti-HBc. The prevalence of IgG antibodies against hepatitis A was 46.0%. The implications in terms of control are discussed.     

Evaluation of the indirect haemagglutination test in the diagnosis of amoebiasis

The IHA test was evaluated in the diagnosis of amoebiasis. Axenically-grown E. histolytica was used as an antigen. A total of 427 sera from symptomatic (intestinal and extra-intestinal) amoebiasis patients. asymptomatic carriers, patients with parasitic intestinal infections other than E. histolytica, and healthy controls were tested. From 288 symptomatic cases of amoebiasis, 232 (80.6 per cent) gave positive reactions. In 93 asymptomatic cases of amoebiasis, 55 (59.1 percent) were seropositive. In testing of sera from 16 subjects with parasitic intestinal infections other than E. histolytica, a low-level positive IHA titres occurred in 2 (12.5 per cent). The test was also positive with a low titre in 3 (10.0 per cent) of the 30 subjects of the healthy control group. The results indicate that the IHA test is of value in the confirmation of intestinal and extra-intestinal amoebic infections especially in cases where the parasite is difficult to demonstrate.     

Diagnosis of abortion and fetal death by the ultrasonographical device in cynomolgus monkeys under indoor individually-caged conditions

This paper describes the rate of naturally occurring abortion and fetal death that were diagnosed by the use of an ultrasonographical device in 683 pregnant cynomolgus monkeys (Macaca fascicularis) of which 542 were wild-imported monkeys and 141 were colony-bred monkeys. All of these animals had become pregnant by the 3-day timed mating system. The combined rate of abortion and fetal death was 7.0% in the wild-imported animals and 7.8% in the colony-bred animals. There were no significant differences in the rate with respect to different ages and to different period of feeding at Tsukuba Primate Center. Seventy-eight per cent (38 monkeys) of the total cases occurred during the first 5 to 9 pregnancy-weeks.     

Efficacy model for antibody-mediated pre-erythrocytic malaria vaccines

Antibodies to the pre-erythrocytic antigens, circumsporozoite protein (CSP), thrombospondin-related adhesive protein (TRAP) and liver-stage antigen 1, have been measured in field studies of semi-immune adults and shown to correlate with protection from Plasmodium falciparum infection. A mathematical model is formulated to estimate the probability of sporozoite infection as a function of antibody titres to multiple pre-erythrocytic antigens. The variation in antibody titres from field data was used to estimate the relationship between the probability of P. falciparum infection per infectious mosquito bite and antibody titre. Using this relationship, we predict the effect of vaccinations that boost baseline CSP or TRAP antibody titres. Assuming the estimated relationship applies to vaccine-induced antibody titres, then single-component CSP or TRAP antibody-mediated pre-erythrocytic vaccines are likely to provide partial protection from infection, with vaccine efficacy of approximately 50 per cent depending on the magnitude of the vaccine-induced boost to antibody titres. It is possible that the addition of a TRAP component to a CSP-based vaccine such as RTS,S would provide an increase in infection-blocking efficacy of approximately 25 per cent should the problem of immunological interference between antigens be overcome.     

Regional distribution of homocarnosine and other ninhydrin-positive substances in brains of malnourished monkeys

—Eight male monkeys (Macaca nemestrina) aged 6–9 months were divided into two groups and fed either an adequate protein diet (20% casein) or a protein deficient diet (2% casein). After 3- 5 months of receiving the low protein diet, the malnourished monkeys showed extensive fatty metamorphosis of the liver cells, distorted patterns of plasma and hepatic free amino acid pools, and other features consistent with the diagnosis of protein-calorie malnutrition. Examination of the cerebrum, cerebellum and brain stem in the malnourished animals revealed profound accumulation of 3-methylhistidine, histidine and homocarnosine in all three regions. For histidine, the cerebral, cerebellar and brain stem levels in the protein deficient animals increased by 145, 104 and 101 per cent over levels observed in corresponding regions of the brain in well-fed monkeys. Similarly, there were significant elevations in homocarnosine contents of the cerebrum (+ 99 per cent), cerebellum (+ 140 per cent) and brain stem (+ 146 per cent) in comparison to levels in control animals. In contrast, the levels of valine, serine and aspartic acid were markedly reduced in all three brain areas in the malnourished animals. Protein-calorie deficiency also produced reductions in the brain levels of taurine, glutamic acid, isoleucine, leucine and threonine which varied in magnitude in the three major regions of the brain examined. These biochemical alterations which may in part underlie some of the psychomotor changes often observed in protein-calorie malnutrition, were discussed not only in relation to the role of amino acids as precursors for the synthesis of neuroregulatory substances but also with due regard to the possibility that some of these ninhydrin-positive substances such as GABA, homocarnosine, glycine and the dicarboxylic amino acids may possess neuroexcitatory or inhibitory properties in various parts of the central nervous system.     

A retrospective study of 932 second trimester terminations using gemeprost (16,16 dimethyl-trans delta 2 PGE1 methyl ester).

A retrospective study of 932 second trimester terminations between 12-27 weeks gestation was carried out to determine the efficacy of gemeprost for second trimester termination. A single course of 5 x 1 mg gemeprost pessaries was administered every three hours. If abortion had not occurred after the first course of pessaries, a further course of 5 x 1 mg pessaries was administered. Intravenous oxytocin was administered after 36 hours if abortion had not occurred. Eighty per cent and ninety five per cent of patients aborted within 24 and 48 hours respectively. Of the remaining 5 per cent of women, 3 per cent aborted with escalating doses of oxytocin. In the remaining 18 (2 per cent) women, the pregnancies were electively terminated with an alternative method. The median induction-abortion interval was 18.0 hours and 15.0 hours in nulliparous and parous women respectively (P less than 0.0001). The number of pessaries required to induce abortion was not influenced by parity. Significantly more parous women bled more than 500 ml. The incidence of pelvic sepsis (0.1 per cent) and cervical tear (0.1 per cent) was low. Twenty six per cent of women had diarrhoea and 23 per cent vomited following administration of prostaglandin. This study confirmed the efficacy of gemeprost for second trimester termination of pregnancy. This method of termination is safe, non-invasive, simple and has a low complication rate.     

Transmission of B virus infection between monkeys especially in relation to breeding colonies

Studies of B virus (Herpesvirus simiae) antibody in several species of macaque lead to the following generalizations. Newborn monkeys are not infected with B virus, even when born of seropositive mothers. Young monkeys remain uninfected until they become adults. The majority of adults develop B virus antibody unless their physical contact with seropositive adults is restricted. These observations are consistent with sexual transmission of B virus and classification of the disease in monkeys as venereal. However, infection at oral and dermal sites also occurs and may play a part in monkey-to-monkey transmission. Epizootics of B virus occurred during early attempts to start B virus-free breeding colonies. They appeared to originate from reactivated latent B virus in adult monkeys which had only low titres of antibody. The stress produced when groups of adult strangers were assembled to form breeding colonies was the most effective known inducer of latent B virus. Total exclusion of animals with any trace of antibody has enabled the establishment of new breeding colonies which are free from B virus.     

Thoracic force-deflection studies in living and embalmed primates

In order to evaluate the validity of the use of the cadaver in making estimates of the living human response to cardio-thoracic impact a series of tests were performed under similar conditions comparing the force-deflection response of the anaesthetized and the embalmed Rhesus monkey. Twelve primates (100 tests) were subjected to controlled thoracic impact with a Remington Humane Stunner while seated in a sled held firmly in place. Tests on 8 of the monkeys were performed while they were anaesthetized and again 30 days after being embalmed. Four monkeys were tested only after embalming. X-rays were used to evaluate rib fractures.

Results showed clear significant differences between anaesthetized primates and those tested 30 days after being embalmed. Forces developed under the same test conditions were 132 per cent higher for the embalmed primates compared to anaesthetized monkeys under static test conditions at maximum displacement and 25–50 per cent higher under dynamic testing conditions for all but the initial part of the tests. The results of these studies indicate that the mechanical characteristics of the primate thorax are dramatically affected by death and/or subsequent embalming procedures.     

Relationship between protein and proteolytic activity in the midgut of mosquitoes.

In female mosquitoes anal injections (enemas) of nutrient solutions were administered in measured amounts to allow direct comparison of protease activities.The amount of protein ingested had a pronounced effect upon the rate of protein digestion, but had little influence upon the rate of protease secretion. Maximal protease activity increased only slightly with increasing meal size and always coincided with the digestion of about 80 per cent of the protein ingested.The use of the enema technique provided an experimental means to reject clearly a neural stimulus for protease secretion. Proof is given for a secretagogue stimulus: the presence of globular proteins with a minimal molecular weight is required for protease secretion.Despite antitryptic factors which are known to occur in different titres in vertebrate bloods, no inhibition was observed in vivo when proteases were recorded after enemas of blood or plasma from several hosts into Aedes aegypti, Anopheles quadrimaculatus, and Culex pipiens quinquefasciatus.Mosquito trypsin was shown to account for about 75 per cent of the proteolytic activity in midgut homogenates. Chymotrypsin was present although with very low activity. Calcium did not stimulate mosquito trypsin as it does mammalian trypsin. Between 22 and 32°C a Q₁₀ of 2·0 was observed for proteases as well as for protein digestion.     

HYPOTHYROIDISM IN MEN IN INDUSTRY—A Preliminary Report on Chemical Evidence

Of 2,807 specimens from blood donors and men in industry, 340 or 12 per cent had serum protein-bound iodine values of 4.0 micrograms or less per 100 cc.In a selected group of 610 “white collar” workers, 52 or 8.5 per cent had similar low values.Careful reexamination of a sample of the latter group indicated that at least 75 per cent of them had hypothyroidism needing treatment.Further analysis of the factors producing hypothyroidism and the validation of the incidence found is warranted. If the incidence observed in this study is confirmed, these findings indicate that 6 per cent to 9 per cent, or more than a million middle aged American men have hypothyroidism.     

Serologic diagnosis of syphilis; the use of Treponema pallidum immobilization and immune adherence tests

TPI tests were carried out on 4,060 sera. Among 3,934 patients with reactive STS and no history of syphilis, 2,148 or 54.6 per cent had negative reaction to TPI tests, 1,695 or 43.1 per cent had positive reaction and 91 or 2.3 per cent had doubtful reaction. Two hundred and ninety-two or 73.0 per cent of 400 pregnant women with reaction to STS in the absence of a history of syphilis showed negative results by TPI test, 103 or 25.8 per cent had positive results and five or 1.2 per cent had doubtful reaction.Ninety-five or 75.4 per cent of 126 patients with a history of treated syphilis had positive reaction to TPI tests, 20 or 20.2 per cent had negative reaction and nine or 9.1 per cent had doubtful reaction.TPI and TPIA tests were done on 143 sera carefully selected for the study. Among 102 sera subjected to the TPI test, 46 or 100 per cent of these positive were also positive by TPIA tests, while 52 or 94.5 per cent of 55 TPI-negative sera were also nonreactive by TPIA test. One serum gave doubtful TPI test reaction and positive TPIA test reaction.     

SEROLOGIC DIAGNOSIS OF SYPHILIS—The Use of Treponema Pallidum Immobilization and Immune Adherence Tests

TPI tests were carried out on 4,060 sera. Among 3,934 patients with reactive STS and no history of syphilis, 2,148 or 54.6 per cent had negative reaction to TPI tests, 1,695 or 43.1 per cent had positive reaction and 91 or 2.3 per cent had doubtful reaction.Two hundred and ninety-two or 73.0 per cent of 400 pregnant women with reaction to STS in the absence of a history of syphilis showed negative results by TPI test, 103 or 25.8 per cent had positive results and five or 1.2 per cent had doubtful reaction.Ninety-five or 75.4 per cent of 126 patients with a history of treated syphilis had positive reaction to TPI tests, 20 or 20.2 per cent had negative reaction and nine or 9.1 per cent had doubtful reaction.TPI and TPIA tests were done on 143 sera carefully selected for the study. Among 102 sera subjected to the TPI test, 46 or 100 per cent of these positive were also positive by TPIA tests, while 52 or 94.5 per cent of 55 TPI-negative sera were also nonreactive by TPIA test. One serum gave doubtful TPI test reaction and positive TPIA test reaction.     

RELATION OF A STREPTOCOCCUS TO EPIDEMIC POLIOMYELITIS-Studies in Etiology,Diagnosis and Specific Treatment

The production in 1915 of herpes zoster or "posterior" poliomyelitis in animals with a streptococcus led to further research on the etiologic importance of streptococci in "anterior" poliomyelitis.A specific streptococcus was demonstrated consistently in persons with poliomyelitis and in well persons having contact with them or merely inhabiting an area in which poliomyelitis was epidemic. That the organism was not present in areas remote from contact with the disease was likewise demonstrated.The streptococcus has been isolated from filtrates of poliomyelitis virus and from the tissues and exudates which harbor the virus. It appears in the spinal fluid in the preparalytic stage of poliomyelitis and disappears from the spinal fluid during the severe stage of the disease. Antibody and antigen prepared from the streptococcus were used to determine the presence of antigen and antibody indicative of streptococcal infection in many patients with poliomyelitis and in well persons. The intensity of reaction indicating specific streptococcal antigen was directly proportional to the degree of paralysis in patients; the reaction was greater in persons whose age, sex and previous isolation from the disease would normally indicate greater susceptibility. The test for antibody gave opposite results. Specific agglutinins for the streptococcus and neutralizing antibody for the virus were present consistently in the serum of persons and monkeys during recovery from poliomyelitis. Virus produced in vitro from the associated streptococcus caused all the clinical and pathologic features of poliomyelitis in monkeys inoculated with it, and the animals that recovered from the disease thus induced were proved to be immune thereafter to the natural virus. Antistreptococcic serum prepared in horses was used to treat poliomyelitis. In a group of monkeys inoculated with the virus of the disease, 6 per cent of those receiving the serum before inoculation died of the disease; of the control group, 82 per cent. In a series of poliomyelitis patients treated with the serum the mortality rate was 8 per cent; in a control series, 21 per cent. In a series treated in all stages of the disease by the author, 10 per cent died; of those who did not receive the serum, 25 per cent. An antibody has been prepared from the streptococcus which appears to prevent paralysis and otherwise mitigate poliomyelitis and to provide immunization from the disease.The conclusion is reached that the virus of poliomyelitis is a form of the specific streptococcus, which is the agent in primary infections and in the development of the immunizing antibody.     

Hypometabolism in Aotus trivirgatus (Primates, Plathyrhini, Cebidae)]

The standard metabolism of Aotus trivirgatus (Night monkey, Owl monkey) is 22.5 to 46.2 per cent below Kleiber's prevision curve for mammals, which applies to other cebid monkeys like Saimiri sciureus and Alouatta. However the metabolic rate of Aotus is not reduced to the extent found in two hypometabolic prosimians Perodicticus potto and Nycticebus coucang. The low metabolism in Aotus is associated with a normal body temperature and a thick fur of high insulating power. These results are discussed.     

RELATION OF A STREPTOCOCCUS TO EPIDEMIC POLIOMYELITIS—Studies in Etiology,Diagnosis and Specific Treatment

The production in 1915 of herpes zoster or “posterior” poliomyelitis in animals with a streptococcus led to further research on the etiologic importance of streptococci in “anterior” poliomyelitis.A specific streptococcus was demonstrated consistently in persons with poliomyelitis and in well persons having contact with them or merely inhabiting an area in which poliomyelitis was epidemic. That the organism was not present in areas remote from contact with the disease was likewise demonstrated.The streptococcus has been isolated from filtrates of poliomyelitis virus and from the tissues and exudates which harbor the virus. It appears in the spinal fluid in the preparalytic stage of poliomyelitis and disappears from the spinal fluid during the severe stage of the disease.Antibody and antigen prepared from the streptococcus were used to determine the presence of antigen and antibody indicative of streptococcal infection in many patients with poliomyelitis and in well persons. The intensity of reaction indicating specific streptococcal antigen was directly proportional to the degree of paralysis in patients; the reaction was greater in persons whose age, sex and previous isolation from the disease would normally indicate greater susceptibility. The test for antibody gave opposite results.Specific agglutinins for the streptococcus and neutralizing antibody for the virus were present consistently in the serum of persons and monkeys during recovery from poliomyelitis.Virus produced in vitro from the associated streptococcus caused all the clinical and pathologic features of poliomyelitis in monkeys inoculated with it, and the animals that recovered from the disease thus induced were proved to be immune thereafter to the natural virus.Antistreptococcic serum prepared in horses was used to treat poliomyelitis. In a group of monkeys inoculated with the virus of the disease, 6 per cent of those receiving the serum before inoculation died of the disease; of the control group, 82 per cent. In a series of poliomyelitis patients treated with the serum the mortality rate was 8 per cent; in a control series, 21 per cent. In a series treated in all stages of the disease by the author, 10 per cent died; of those who did not receive the serum, 25 per cent.An antibody has been prepared from the streptococcus which appears to prevent paralysis and otherwise mitigate poliomyelitis and to provide immunization from the disease.The conclusion is reached that the virus of poliomyelitis is a form of the specific streptococcus, which is the agent in primary infections and in the development of the immunizing antibody.     

Antibiotic sensitivity and pathogenetic factors of hospital strains of Pseudomonas aeruginosa isolated from patients treated in a resuscitation unit]

Strains of Pseudomonas aeruginosa were isolated from patients treated in the Centre of Thermal Affections in 1985-1989. It was shown that 72.9, 59.3, 33.8 and 54.2 per cent of the isolates were sensitive to cefotaxime, tobramycin, gentamicin and polymyxin, respectively. The study of pathogenicity factors of the isolates revealed that 83 per cent of the strains produced thermolabile enterotoxin, 79.6 per cent of the strains had adhesive activity and 71.1 per cent of the strains produced hemolysin. The study detected combinations of various pathogenicity factors. 42.3 per cent of the isolates had both adhesive and enterotoxigenic properties. Adhesiveness and hemolytic activity were shown by 13.5 per cent of the strains. 16.9 per cent of the strains produced both enterotoxin and hemolysin. Adhesive activity, enterotoxigenicity and hemolysin production were observed in 6.7 per cent of the strains. It was noted that the strains of P. aeruginosa resistant to polymyxin mainly produced enterotoxin (18.6 per cent) and those resistant to cefotaxime had adhesive activity (34.0 per cent).     

INFLUENZA VACCINE,ASIAN STRAIN—Reactions Following Its Use in Adults

A study of reactions following influenza vaccine inoculation of 327 employees of Peralta Hospital, 55 men and 272 women, showed a very low value for significant or severe reactions. The reaction rate as observed with the present monovalent vaccine containing 200 CCA units of Asian strain, Type A influenza virus, was considerably lower than that reported with previous polyvalent vaccines containing up to 1,400 or 1,500 CCA units of total virus content.The absenteeism rate was 1.1 per cent for women, nil for men.The incidence of reactions was much greater in women than in men. Local reactions such as pain, swelling, or redness at the site of injection occurred in 29.1 per cent of men and 35.7 per cent of women. The incidence of systemic reactions—fever, aching, chilliness, headache, nausea and vomiting—was 3.6 per cent in men and 8.8 per cent in women. About 9 per cent of men and 30 per cent of women had both local and systemic reaction. Some 58 per cent of men and 25 per cent of women had no reaction.The greater majority of reactions appeared within five hours after inoculation with influenza vaccine.In adults the prevention of anaphylactic reactions due to the small amount of egg protein in influenza vaccine, can be accomplished by screening for history of hypersensitivity to egg, chicken or chicken feather. In questionable cases, intradermal testing can be done.The reaction rate observed in this study for the present influenza vaccine was so low that it ought not deter immunization.     

Effect of the oxidation-reduction potential of the medium on the production of gentamycin

The medium redox potential had an effect on gentamicin production by Micromonospora purpurea v. violacea, strain VNIIA 7R. The Eh influence was shown to be statistically reliable when the results were expressed in relative units against the control. In the laboratory experiments with low volumes of the medium the Eh increase by more than 170 per cent induced inhibition of gentamicin biosynthesis while the Eh increase by 108 to 168 per cent induced stimulation of the activity.