pH-independent inhibition of plasma angiotensin I degradation: implications for renin assay. Valid enzymatic assay of plasma renin (EC 3.4.99.19) requires complete inhibition of enzymes that destroy angiotensin I. Historically, the degree of such inhibition has varied with assay pH and with the combination of protease inactivators employed. To circumvent this problem, we studied the effects of several protease inactivators on both the formation and degradation of angiotensin I by human plasma at pH 5.5 and at pH 7.4. While several reagent combinations effectively suppressed the rate of angiotensin I disappearance at one of the two pH values, three combinations of phenylmethylsulfonyl fluoride with chelating agents accomplished this at both pH values. Of these, however, only the combination of 5 mmol/l ethylenediamine tetraacetate, 1.5 mmol/l 8-hydroxyquinoline, and 7.5 mmol/l phenylmethylsulfonyl fluoride also maximized the rate of angiotensin I formation at both pH 5.5 and 7.4. Thus, the use of the latter reagent combination would facilitate comparison of renin assays performed at different pH values and would permit standardization of the protease inactivators employed in such assays.


Longitudinal study of circulating immune complexes in a patient with Staphylococcus albus-induced shunt nephritis. The direct measurement and partial characterization of circulating immune complexes has been performed in a longitudinal study of a patient with Staphylococcus albus-induced shunt nephritis. The high levels of immune complexes were associated with cryoglobulinaemia and hypocomplementaemia. The activation of complement was found to be via the classical pathway, but the functioning of the alternative pathway may have been impaired in vivo due to very low levels of C3. The host response to the infection was also characterized by the production of a marked macroglobulinaemia, high titres of rheumatoid factor and a typical acute phase increase in the C-reactive protein level. Immune complex levels were persistently elevated many months after the removal of the focus of the infection. A possible explanation for this surprising finding may lie in the nature of the antigens in the immune complexes. It was found that the immune complexes contained both antibodies to and antigens from Staphlococcus albus. In particular, glycerol teichoic acid and staphylococcal nuclease were identified as components of the immune complexes present during the acute phase. Glycerol teichoic acid was also identified in the immune complexes found later although other Staphylococcus albus antigens as yet unidentified were also present and persisted in the circulation for several months.


An IgM monoclonal protein with multiple serological specificities. An IgM lambda M-component with a false Wasserman reaction and 'false' rheumatoid factor activity from a patient suffering from a well-differentiated lymphocytic lymphoma is presented. The monoclonal protein showed antibody activity against cardiolipin and cross-reacted with acryl particles, not with human IgG. Both these activities were found in the Fab fragment. The reduction of the 19S IgM to 7S IgM subunits was responsible for a strong decrease in activity. The importance of the IgM quaternary structure in determining antibody affinity is emphasized.


Celluar immunoglobulins in human gamma- and alpha-heavy chain diseases. Proliferating cells from twenty-four patients with alpha- or gamma-heavy chain disease (HCD) were studied by direct immunofluorescence and in several cases by biosynthesis experiments with 14C-amino acid incorporation. In twenty-two patients, the cells contained the HCD proteins only and no light chain synthesis could be detected. Conversely, apparently non-secreted monotypic light chains were found in one case of gamma-HCD and one case of alpha-HCD. The proportion of proliferating cells containing cytoplasmic heavy chains, their appearance and the presence or not of surface heavy chains showed great variation from patient to patient. In some cases, the proliferation predominantly affected either plasma cells or lymphocytes whereas in others the disease seemed to correspond to a proliferation of HCD protein-bearing lymphocytes with persistent maturation into plasma cells. Large cell lymphomas supervening on alpha-HCD belonged to the same proliferating clone as the clone secreting the HCD protein, as shown by surface markers and biosynthesis experiments which demonstrated synthesis but no secretion of HCD proteins. In one patient with gamma-HCD, the cells carried surface gamma and delta chains.


Metabolism of carbaryl by kidney, liver, and lung from human postembryonic fetal autopsy tissue. Metabolic profiles of carbaryl in human postembryonic fetal autopsy tissue were determined using an in vitro tissue-maintenance technique. 1-Naphthyl-14C or N-methyl-14C-carbaryl was applied to growth medium containing explants of the tissue. Each mixture was incubated for 18 hr and the medium analyzed by DEAE-cellulose column chromatography. Fetal liver performed the metabolic processes of demethylation, hydrolysis, hydroxylation, and oxidation, followed by conjugation, as was found with adult liver. However, the anionics from fetal liver amounts to 20% of those found with adult liver. The kidney made naphthyl glucuronide and naphthyl sulfate, whereas the lung produced naphthyl sulfate from carbaryl. The metabolic activities of the fetal kidney and lung were close to the corresponding human adult tissues based upon the anionic metabolites found and the amount of unmetabolized carbaryl in the medium after 18 hr of incubation. Silica gel chromatography of ether-extractable neutral fractions from DEAE-cellulose revealed 3,4, and 9 ether-extractable metabolites from lung, kidney, and liver, respectively. The present study shows that the in vitro technique is capable of semiquantitatively demonstrating the metabolic activities of specific organs from the human fetus.


The subcellular distribution of gold in monkey liver, kidney and spleen, in vivo. 1. The subcellular distribution of gold in the livers, kidney cortex and the spleens of rhesus monkeys was determined at 2, 10 and 20 days after the administration of sodium (195Au)-aurothiomalate. The serum gold levels were also determined over the period of study. 2. With the continued decrease in the serum gold levels from day 1 to day 20, a continued increase in the tissue levels of the organs studied was observed. The corresponding subcellular fractions of the tissues also showed continued increases in the radioactive concentrations of gold. 3. Gold was found to localise in all the organelles of the cells in the liver, kidney cortex and the spleen, though in varying amounts. The largest increases in the concentrations of gold from day 2 to day 20 were found in the lysosomal bodies of liver and kidney. The concentrations in the kidney cortex increased from approximately five times those in the liver or the spleen on day 2 to about eight times those on day 20.


Bacterial plasmids: autonomous replication and vehicles for gene cloning. The use of recombinant DNA techniques in the analysis of the structure and replication of bacterial plasmids has provided much information on the properties of these genetic elements and has led to the construction of plasmid elements that are potentially very useful as gene cloning vehicles in Escherichia coli and other gram-negative bacteria. The genetic and molecular properties of plasmids mini-F, ColE1, and RK2 are described with particular emphasis on the origin and direction of replication and the identification of genetic regions essential for maintenance of these elements in the extra-chromosomal state. Low molecular weight derivatives of each of these plasmids have been obtained and a restriction enzyme map determined for these various derivatives. A hybrid DNA molecule consisting of a low molecular weight derivative of ColE1 joined to a segment of bacteriophage DNA has been constructed and shown to be capable of existing either as a plasmid element or packaged as an infectious viral particle. Finally, several of the low molecular weight derivatives of these plasmids described have certain advantages as vehicles for the cloning of DNA including derivatives of he broad host range plasmid RK2 that may be useful for gene cloning in gram-negative bacteria distantly related to E. coli.


Recurrent infective endocarditis in a drug addict. Multiple separate episodes in nine years. Multiple episodes of documented recurrent infective endocarditis occurred in an habitual drug abuser. The six recurrences represent the largest known number reported in a single patient. The case illustrates many characteristics of infective endocarditis and serves to emphasize that recurrent infections are likely if endocarditis risk factors are not eliminated.


Radiation therapy and mechanical dilation of endobronchial obstruction secondary to Wegener's granulomatosis. A patient with Wegener's granulomatosis, which was otherwise well controlled with steroids and cyclophosphamide, had persistent total obstruction of an inflamed left mainstem bronchus. The inflammation resolved, and the lung expanded following radiation therapy. However, intermittent atelectasis and pneumonia occurred distal to a residual short stricture. Repeated dilation endoscopically with Plummer bougies has proven effective in maintaining good ventilation and preventing recurrence of the atelectasis and pneumonia in the left lung for 18 months.


Preparation of highly potent Naja naja atra (Formosan cobra) antivenin. As the preparation of high titer antiserum against Naja naja atra venom is a time-consuming process, attempts were made to develop a immunization procedure for producing highly potent antiserums within a short period. Rabbits were immunized for 12 weeks with (1) whole venom as used routinely in our Institute, (II) whole venom adsorbed on carboxymethyl-cellulose (CMC) and mixed with Freund's complete adjuvant and (III) neurotoxin adsorbed on CMC and adjuvant mixture followed by CMC-whole venom-adjuvant mixture. The results showed that one ml of the antiserum prepared by method (III) could neutralize 445 LD50 of whole venom, in other words, its potency was 4 and 40 times higher than those prepared by methods (II) and (I), respectively.


The nature and role of pregnancy-associated antigens and the endocrinology of early pregnancy in the ewe. Depite descriptions of many of the physiological events of early pregnancy in the ewe, the way in which the immunological and endocrine adjustments are brought about is not well understood. Obviously, specific effects--for example, maintenance of the corpus luteum--are brought about by the presence of a conceptus in the uterus. This has led to a search for substances produced by the conceptus or induced by the presence of the conceptus in the uterus which could be involved in establishing pregnancy. This paper describes the presence in sheep of substances associated with pregnancy and discusses the origin and characterization of these substances and their role in ovine pregnancy.


Comparative aspects of blastocyst-endometrial interactions at implantation. Since the trophoblast-uterine adhesion is as nearly a universal phenomenon in implantation as can be found, an attempt was made to determine whether or not there was a reduction in cell surface glycoproteins in the rat, as can be observed in the ferret. Neither colloidal iron nor cationized ferritin revealed the type of pattern anticipated for a localized reduction in surface negativity in the imprint of the blastocyst in the implantation chamber. The use of lectin-coated latex beads also proved disappointing in defining regional differences in adhesiveness. However, a number of observations on the changing shape of the implantation chamber, the secretion of periluminal material by decidual cells, and the penetration of the residual basal lamina of the luminal epithelium by the decidual cells were made in the course of these studies. The implantation chamber of the rabbit, in which the blastocyst does not make an imprint, was contrasted with that of the rat. The areas of fusion of trophoblast knobs with uterin epithelial cells shown to be visible by scanning electron microscopy. Finally, some observations on the hypertrophy of maternal epithelial cells to form the uterine plaque in the rhesus monkey are described, and the hypertrophy of endothelial cells to form admirably suited to protein secretion is presented.


Immunological interference with the maternal recognition of pregnancy in primates. If chorionic gonadotropin is involved, as part of the luteotrophic stimulus, in the maternal recognition of pregnancy in primates, then immunization against the beta subunit of hCG should prevent a conception going beyond the length of the normal cycle. Immunization against hCG-beta suppresses fertility in marmoset monkeys, baboons and rhesus monkeys, as long as the antibody titres remained high. As titres decline, however, female marmosets experience recurrent abortions interspersed with occasional live births. Young marmosets born to mothers with low circulating levels of antibody reach apparently normal sexual maturity. Immunization against hCG-beta subunit may have potential as a possible long-term method of fertility control in humans, but its reversibility needs further study. There is still a lack of basic knowledge of the functions of chorionic gonadotropin, especially now that its restriction to pregnancy seems improbable. Immunization against hCG may be valuable as a research tool with which to study the immunological relationships between mother and fetus during early pregnancy.


Deoxyribonucleoside toxicity in adenosine deaminase and purine nucleoside phosphorylase deficiency: implications for the development of new immunosuppressive agents. The immunodeficient state associated with adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) deficiency may result from the selective phosphorylation by thymus-derived lymphocytes of the ADA substrate deoxyadenosine and the PNP substrate deoxyguanosine, leading to the intracellular trapping of toxic deoxyribonucleoside triphosphates. Agents such as deoxycytidine might be able to favourably modify the immunodeficient state by inhibiting deoxyribonucleoside phosphorylation. Deficiencies of other nucleotide catabolic enzymes, if selectively expressed by lymphocytes, might also lead to immunodeficiency via nucleoside trapping in lymphoid tissues. Purine deoxyribonucleoside analogues, either alone or in combination with ADA inhibitors, may have value as lymphospecific antimetabolites.


Purine nucleoside phosphorylase: immunodetection and characterization of the human enzyme. An antibody prepared against human placental purine nucleoside phosphorylase (PNP) has been used to characterize the subunit structure of the normal enzyme by two-dimensional gel electrophoresis. The antibody against the placental enzyme reacts with PNP present in human lymphocytes, fibroblasts, erythrocytes and placenta. The enzyme can be detected in intact cells by immunofluorescence with the same antibody preparation. The techniques allow the identification of cross-reacting material in a small number of lymphocytes and could therefore be used to detect abnormal enzyme protein in immunodeficiency.


An approach to the restoration of T cell function in a purine nucleoside phosphorylase deficient patient. A patient with a selective impairment of T cell-dependent immunity based on a purine nucleoside phosphorylase (PNP) deficiency has been treated with transfusions of irradiated erythrocytes and plasma. After each transfusion with PNP-containing erythrocytes a decrease in accumulated nucleosides and their deoxy compounds was observed, whereas uric acid excretion and serum uric acid increased. Lymphocyte counts increased transiently after each erythrocyte and plasma infusion and a partial restoration of T cell-dependent immunity was gradually attained. The pattern of restoration was reminiscent of the immunological reconstitution seen in patients with severe combined immunodeficiencies treated with bone marrow transplantation. Amelioration of T cell-dependent immunity was shown to be related to the metabolic changes. On the basis of the presumed mechanism of lymphocyte intoxication and consequently starvation of intracellular DNA precursors, deoxycytidine was given orally. This did not lead to further improvement in immunological function. However, partial restoration of immunological disturbances in PNP deficiency can be attained by erythrocyte transfusions and evidence is presented that additional pharmacological approaches are possible.


[Epidemiology and prognosis of non-A, non-B hepatitis (author's transl)]. The frequency of non-A, non-B hepatitis (n = 325) was determined among all cases (n = 1368) of acute viral hepatitis observed in the Hannover are abetwen 1975 and 1978. Hepatitis A was excluded by demonstration of anti-HAV-IgM, hepatitis B by demonstration of HBs antigen or an isolated occurrence of anti-HBc at the beginning of the disease. Non-A, non-B hepatitis occurred predominantly in adults and showed no seasonal variability. As a consequence of results of followup investigations in 174 hepatitis patients 2 years after the onset of the disease it can be assumed that non-A, non-B hepatitis tends to lead to chronic courses more frequently than hepatitis B.


[Mucopolysaccharidosis I-S (Scheie's disease) (author's transl)]. Mucopolysaccharidosis I-S in two encygotic twin sisters. The disease is characterised by opacification of the cornea, hepatospenomegaly, carpal tunnel syndrome, flexion contractures of the fingers and toes, valvar heart disease, normal intelligence and essentially normal behaviour. The biochemical defect consists of deficiency of the lysosomal hydrolase alpha-L-iduronidase. Urinary dermatan-sulphate excretion is raised. The muscles are also involved:the electron microscopic revelas interstitial storage of glycosaminoglycan-like substance. There is no specific treatment.


Potentiation by indomethacin of TRH-induced TSH secretion in the rat. We have studied the effect of two inhibitors of prostaglandin synthesis on the basal and TRH-stimulated plasma TSH levels in the rat. Animals were injected sc daily with indomethacin 3 mg/0.5 ml) or aspirin (16--30 mg/0.5 ml) for 3 days. The plasma T4 and T3 were consistently lower in the indomethacin or aspirin groups than in the controls, while the basal TSH levels did not change. Indomethacin treatment significantly potentiated the TSH response to synthetic TRH (20 ng. iv) in intact and thyroidectomized rats. The pituitary TSH content was markedly increased by indomethacin, while hypothalamic TRH content did not change. In contrast, aspirin inhibited the TSH response to TRH in intact rats, when pituitary TSH content decreased significantly. No potentiation by aspirin of TRH-stimulated TSH response in the thyroidectomized rats was observed. The increased sensitivity of plasma TSH response to exogenous TRH in the indomethacin group is presumably due to higher pituitary TSH content than in the controls. The action of indomethacin appears to be mediated, at least in part, at the pituitary level. In addition, there is a dissociation between the action of indomethacin and the action of aspirin in the TSH response to TRH.


The pattern of urinary excretion of corticosteroids in Rhesus monkeys (Macaca mulatta). Urinary total 17-oxogenic steroids (17-OGS), cortisol, cortisone, corticosterone, tetrahydrocortisol (THF), allo-tetrahydrocortisol (all-THF), tetrahydrocortisone (THE), cortols and cortolones, were estimated by established methods in 30 female and 20 male rhesus monkeys. The pattern of the excretion of these steroids in this species was comparable with the human corticosteroids excretion, irrespective of sex difference. The results obtained from this investigation show that they could be used during the study of adrenocortical function and cortisol metabolism in this species.


Decrease of plasma somatotropin levels following thyroliberin (TRH) administration in patients with hyperthyroidism. Thyroliberin was observed to induce a significant decrease of plasma somatotropin levels in patients with untreated hyperthyroidism, except those with very low initial somatotropin levels. These data have confirmed our previous finding where we noted that under the conditions of thyroid hormone excess thyroliberin was able to exert an inhibitory effect on somatotropin release.


Recording small sharp spikes with depth electroencephalography. Two patients with intractable seizures and focal temporal sharp waves also had small sharp spikes as incidental findings in their scalp electroencephalograms. Depth electroencephalography verified the intracerebral origin of the small sharp spikes and differentiated them from the more significant epileptiform abnormalities.


Relationships between plasma concentrations of diphenylhydantoin, phenobarbital, carbamazepine, and 3-sulfamoylmethyl-1,2-benzisoxazole (AD-810), a new anticonvulsant agent, and their anticonvulsant or neurotoxic effects in experimental animals. The relationships between plasma concentrations of diphenylhydantoin (DPH), phenobarbital (PB), carbamazepine (CBZ), and 3-sulfamoylmethyl-1,2-benzisoxazole (AD-810), a new anticonvulsant agent, and their anticonvulsant and neurotoxic effects were studied in various species of animals. Anticonvulsant activities of test drugs were examined by the maximal electroshock seizure (MES) test. Neurotoxicities were determined by the rotorod performance test in mice and rats and by behavioral observations in rabbits, dogs, and monkeys. It was demonstrated that both the anticonvulsant effects and the neurotoxic effects of the drugs tested were more closely correlated with their plasma concentrations than with the dosages administered. There was a critical plasma concentration for each drug to show an anticonvulsant effect or to cause a neurotoxic effect in an individual animal. The critical plasma concentrations for anticonvulsant and neurotoxic effects of each drug were relatively constant among different species, with the exception of DPH in rabbits, which had twice the value in other species. The therapeutic ranges of plasma concentrations of DPH, PB, and CBZ determined in various species of animals coincided well with those recommended clinically. AD-810 was found to be effective against MES without signs of neurological toxicity in the ranges of plasma concentrations of 9.8 to 74.0, 10.8 to 95.0, 9.6 to 117.0, and 12.6 to 96.2 microgram/ml in mice, rats, rabbits, and dogs, respectively. These results seem to suggest that AD-810 may be effective clinically at plasma concentrations above 10 microgram/ml, with a therapeutic range up to 70 microgram/ml, which is much wider than the therapeutic ranges of DPH (10--20 microgram/ml), PB (10--30 microgram/ml), and CBZ (4--10 microgram/ml).


Effects of chronic cerebellar stimulation on chronic limbic seizures in monkeys. No influences of chronic cerebellar stimulation were found in 10 different controlled experiments in 5 different monkeys with chronic alumina-induced psychomotor seizures. The stimulation parameters used were comparable to those used in human epileptics, and continuous daily EEG and behavioral monitoring allowed all seizures to be measured for daily frequency and duration over the several weeks of the experiments. Nocturnal seizures were similarly quantified in 3 monkeys to verify that cerebellar stimulation did not affect them. Motor cortex potentials evoked by cerebellar pulses confirmed that the stimulations were activating the cerebellum throughout the experiments, and measures of electrode access resistance and impedance verified that the electrodes remained in contact with the cerebellum. In one monkey given phenobarbital medication, interictal morbid behavior appeared to be improved by chronic stimulation of either cerebellum or dorsolateral frontal cortex, thus indicating an arousal influence of brain stimulation not due to cerebellum per se.


A case of Lennox-Gastaut syndrome successfully treated by removal of a parietotemporal astrocytoma. We report a case of Lennox-Gastaut-type epilepsy that was followed for 3 years. The patient showed typical EEG findings and later a parietotemporal astrocytoma (type II) on the left. Removal of the tumor was followed by recovery of the child and disappearance of EEG epileptic elements.


Posttraumatic epilepsy prophylaxis.

Posttraumatic epilepsy prophylaxis. Despite a large body of experimental evidence suggesting that posttraumatic epilepsy can be prevented, there is no generally accepted pharmacological regimen for posttraumatic seizure prophylaxis. This article describes a phenytoin anticonvulsant regimen specifically tailored for the patient with acute head injury and designed to provide immediate and sustained plasma concentrations of phenytoin between 10 and 20 microgram/ml. Initially, an intravenous phenytoin dose of 11 mg/kg body weight is immediately followed by an intramuscular dose of 13 mg/kg body weight. This is followed by daily intramuscular maintenance doses, usually 8.8 mg/kg body weight, until oral medication can be tolerated. Maintenance dosage adjustments, when necessary, are based on serial plasma concentrations of the drug. Eighty-four patients with severe head injuries with substantial risk of posttraumatic epilepsy were administered this regimen. Only 6% of these patients had seizures during the first year after injury (first week excluded), and this is considerably less than the rates reported elsewhere in the literature. Only one-third of these patients are known to have continued to take phenytoin after the first month, and only half of these had plasma phenytoin concentrations above the desired minimal level. The greatly reduced incidence of posttraumatic seizures in these patients, despite the low rate of long-term drug compliance, suggests that a prophylactic effect, rather than a suppressive effect, is produced.


Clonazepam in a focal-motor monkey model: efficacy, tolerance, toxicity, withdrawal, and management. Since the clinical data have been equivocal in regard to the effects of clonazepam (CZP) in focal-motor seizures, an alumina gel monkey model was used to evaluate quantitatively its efficacy with respect to this seizure category. The insolubility of CZP and its short biological half-life in monkey necessitated its evaluation in the model via constant-rate intravenous administration in a solution of polyethylene glycol 400 (PEG). Two groups of monkeys were given CZP in PEG (N = 6) or a PEG solution alone as a control compound (N = 5) for 6 weeks; these treatments were bordered at both ends by 3 weeks of treatment with saline only in order to establish a baseline. CZP was administered at a concentration sufficient to achieve a plasma level of 30 ng/ml in drug step I (3 weeks) and at least double that level in drug step II (3 weeks). As a solute for CZP, and when given by itself, PEG was always administered at a concentration of 35%. The results indicate that CZP is effective for focal-motor seizures and secondarily generalized tonic-clonic seizures, particularly when its concentration in plasma is higher than 60 ng/ml. Withdrawal seizures were evident on cessation of CZP administration. CZP appears to be a useful broad-spectrum anticonvulsant when managed carefully. An unexpected finding was the irreversibility of the pharmacological effect of PEG. Cessation of PEG administration significantly reduced seizure frequency in subsequent weeks to a level below the initial baseline level.


Stupor following administration of valproic acid to patients receiving other antiepileptic drugs. Stupor is an unusual complication following the addition of valproic acid to other antiepileptic drugs. We report four such cases. Stupor occurred acutely in 3 patients and insidiously in the fourth. In the cases of acute toxicity, neither toxic levels of valproate nor significant elevations in blood levels of the other drugs occurred. In the fourth patient, stupor occurred concomitantly with a rise in the phenobarbital level. The electroencephalograms of all 4 patients revealed generalized high-amplitude rhythmic bisynchronous delta activity. Recovery occurred following discontinuation of valproic acid or other antiepileptic drugs. These observations suggest that stupor may occur as a result of drug interactions after the addition of valproic acid to other antiepileptic drugs.


A translocation X;Y system for detecting meiotic nondisjunction and chromosome breakage in males of Drosophila melanogaster. A nondisjunction and chromosome breakage screening system devised by Craymer and modified in our laboratory, involves an X;Y translocation with the short arm of the Y (Ys), marked with the wild type allele of yellow, attached to the distal end of an X (break point 11D) carrying the recessive marker y; and the long arm of the Y chromosome (YL), marked with the dominant locus Bar of Stone (BS), attached to the proximal end of the X. A female tester strain carrying normal chromosomes homozygous for the yellow allele is employed in the mating scheme. Following normal disjunction in the male, all zygotes, which in this case receive aneuploid paternal sex-chromosomes and a normal euploid maternal complement, will die as a result of genetic imbalance. Thus all survivors from this corss can be classified as exceptions arising from: (1) nondisjunction in the female; (2) gross deletion of the paternal X;Y chromosome; (3) complete loss of the paternal X;Y chromosome; or (4) primary meiotic nondisjunction in the male. Results indicate the sensitivity of this scheme for the detection of events induced by x-rays and various chemicals. Positive results have been obtained with the known mutagens EMS and x-radiation.


Studies of human liver bilirubin-glycosyl transferase. Bilirubin UDP-xylosyl and UDP-glucuronyl transferase activities in diseased human livers. The activity of bilirubin UDP-xylosyl transferase as well as UDP-glucuronyl transferase in liver biopsy specimens of 3 control subjects, 42 cases with liver disease and 5 cases with Gilbert's syndrome was measured. Normal values of these enzyme levels were determined to be 142--302 U/kg protein for the former and 260--400 U/kg protein for the latter. Both enzyme levels in acute hepatitis in convalescence and chronic hepatitis were nearly in the normal range. In the cirrhotic liver they tended to a small decrease and patients with Gilbert's syndrome demonstrated significantly decreased enzyme levels. These enzyme levels were only correlated with serum unconjugated bilirubin concentration, but not with the other liver function tests. Finally, both enzyme activities were exactly correlated with each other.


Apolipoprotein A-IV: a protein occurring in human mesenteric lymph chylomicrons and free in plasma. Isolation and quantification. 1. Human mesenteric lymph chylomicrons were isolated from chylous ascites fluid by ultra-centrifugation and agarose/gel chromatography and their apoprotein composition was analysed by dodecylsulfate/polyacrylamide gel electrophoresis, analytical isoelectric focusing and immuno-chemically. Major components of mesenteric lymph chylomicrons were apoprotein A-I, proteins of Mr less than 15 000 including the C-group apoproteins and a protein of Mr 46 000. Minor components were apoprotein E and a protein of Mr approximately equal to 200 000 (B-like protein). This apoprotein composition was qualitatively identical with that of chylomicrons from intestinal lymph of the rat, but was distinctly different from plasma chylomicrons of humans with fasting chylomicronaemia. 2. The protein of Mr approximately equal to 46 000 has been isolated by preparative dodecylsulfate/polyacrylamide gel electrophoresis from human and rat lymph chylomicrons and was compared to a protein of identical Mr present in rat high-density lipoproteins (apoplipoprotein A-IV) and in the rho less than 1.006 g/ml serum lipoprotein fraction of individual humans with alimentary hypertriglyceridaemia. In both species the 46 000-Mr proteins isolated from lymph and serum were identical according to amino acid composition and isoelectric point in 6 M urea. The human proteins from both sources were also immunologically identical. The similarities in the molecular properties of the human apolipoprotein and rat apolipoprotein A-IV indicate that these proteins are homologous. 3. Plasma levels of human apolipoprotein A-IV determined by electroimmunodiffusion were 14.15 +/- 3.66 mg/100 ml (n = 59), but greater than 90% of the protein was unassociated with the major lipoprotein fractions. It is concluded, that apolipoprotein A-IV is a main protein component of human lymph chylomicrons, that is removed from the particles in the plasma compartment.


Studies on the structure of the rabbit kidney brush border. The effects of salts and non-ionic detergents on renal brush borders have been studied. 2 M sodium chloride, iodide or thiocyanate dissociated up to 40% of the protein from the brush borders, destroying the core filaments and resulting in the formation of membrane vesicles; EDTA had a similar effect on structure but released little protein. Triton X-100 and Nonidet P-40 extracted up to 60% of the protein including the major membrane glycoproteins and the enzymes trehalase, maltase and aminopeptidase (microsomal). Triton exhibited a selective effect on lipids removing phosphatidylserine, phosphatidylethanolamine and sphingomyelin but not the bulk of the phosphatidylcholine or cholesterol. The residual structures after Triton extraction comprised the core filaments associated with vesicles of lipid containing alkaline phosphatase and several other proteins. Treatment of these core-vesicle complexes with 2 M sodium chloride dissociated the filaments, releasing the vesicles which could be recovered as a pellicle on centrifugation. It is suggested that the proteins found in the vesicles might serve to interconnect the core filaments with the lipid bilayer.


Effects of apramycin, a novel aminoglycoside antibiotic on bacterial protein synthesis. 1. The novel aminoglycoside antibiotic apramycin is shown to be a potent inhibitor of protein synthesis in bacteria both in vivo and in vitro. 2. In cell-free systems from Escherichia coli programmed with poly(U), apramycin induces translation errors, as assayed by incorporation of leucine, isoleucine and serine, although this effect occurs only to a limited extent. 3. Apramycin inhibits the translocation step of protein synthesis both in vivo, in protoplasts of Bacillus megaterium, and in vitro, in cell-free systems from E. coli. It is proposed that this is the primary inhibitory effect of the drug.


The value of whole body bone scan in the pre-operative assessment in carcinoma of the breast. In 126 patients with primary breast cancer a patient moving whole body bone scan was performed when they first presented. None of the patients in stage I had an evidence of skeletal metastases. Two patients (3%) of 62 in stage II and 4 patients (17%) in stage III had evidence of skeletal metastases. It appears that whole body scanning is the most accurate, sensitive and convenient method of detecting osseous metastases and of staging breast cancer. This investigation should be carried out pre-operatively. Detection of early asymptomatic bony metastases will provide a better planning of treatment with rational approach.


Evaluation of one dimensional profile scans with a whole body counter--82bromide space in liver cirrhosis: concise communication. One dimensional profile scans with a whole body counter are refocused with a linear space invariant filter in the frequency domain, 'Hanning' window and inverse system transfer function. Artifacts are eliminated in a way that a clinically applicable method is developed. Topographic information is achieved with the highly energetic nuclide 82bromide. The bromide space (extracellular volume) in patients with liver cirrhosis (n = 9) compared to normal subjects (n = 10) is increased most in the upper abdominal thoracic region.


Idiotypic cross-reactivity of human and murine phosphorylcholine-binding immunoglobulins. The idiotypic cross-reactivity of mouse and human monoclonal immunoglobulins with binding activity for phosphorylcholine (PC) was investigated, using an idiotypic antibody elicited against the PC-binding human IgMFR. The isolated FR heavy chain proved to be a better inhibitor for the reaction of IgMFR with anti-FR than the FR light chain, but the intact protein was necessary for full idiotypic expression. PC was an inhibitor only at concentrations greater than 10(-3) M indicating that the idiotypic antibody was not combining site-directed. Among the murine PC-binding IgA myeloma proteins, MOPC 167 was found to be the best inhibitor, but its inhibitory capacity was about 4 orders of magnitude lower than that of the homologous IgMFR. McPC 603 was an even weaker inhibitor, while TEPC 15 effected no better inhibition than human monoclonal immunoglobulins without PC-binding activity. MOPC 167 has a most similar binding specificity to IgMFR as indicated by the high affinity for choline of these two proteins. TEPC 15 and McPC 603, on the other hand, exhibit a much lower affinity for choline. In addition to their similarity in specificity, proteins FR and MOPC 167 show important structural similarities within parts of their heavy and light chain variable domains. The data provide some evidence for the existence of idiotypic cross-reactivity between the two species man and mouse.


A mouse myeloma variant with a defect in light chain synthesis. A spontaneous assembly variant, B 50, has been isolated from the MPC-11 mouse myeloma cell line. This variant synthesizes fewer light chains per cell than the parent resulting the production of a slight molar excess of heavy chains. These changes are associated with a delay and change in the pathway of assembly and a delay in secretion. Spontaneous revertants of B 50 have been obtained, all of which synthesize normal amounts of light chains and assemble and secrete the immunoglobulin molecule through the same pathways and with the same kinetics as the parental cells. A comparison of the tryptic-chymotryptic peptides of the parental, variant and revertant heavy and light chains did not reveal any differences. These studies indicate that variants in mouse myeloma cells can arise with defects in the quantitative expression of the immunoglobulin gene and suggest that the presence of excess light chains facilitates the assembly and secretion of some immunoglobulin molecules.


Presence of serum IgD and IgD-containing plasma cells in the mouse. A new technique which makes use of anti-delta-specific allo-or heteroantisera coupled to Sepharose 4 B has been employed to investigate the presence of IgD molecules in the sera of three mouse strains. The sera were reacted with the insoluble immunosorbents, and the eluted material was labeled with 125I prior to antigenic or sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. This material reacted with anti-delta or anti-L chain reagents, but not with other anti-class-specific antisera and, after reduction, was resolved in three discrete bands in SDS-PAGE. The fastest of these bands comigrated with L chain, whereas the other two moved identically to mouse lymphocyte surface delta chains. Immunofluorescence studies on fixed smears of spleen and Peyer's patches' cells revealed the presence of a small but consistent number IgD-containing plasma cells.


Mutations affecting immunoglobulin light chain secretion by myeloma cells. I. Functional analysis by cell fusion. Two clones of MOPC 315 cells have been selected which synthesize but do not secrete Ig lambda light chains. These clones were analyzed by fusion with a cell line synthesizing and secreting kappa chains. Conditions were established for recovery at high frequency (approximately 10(-3)) of spontaneously fused, viable hybrid cells. The resulting hybrid cell lines synthesized both kappa and lambda chains but secreted only kappa chains. Hybrid cells produced by fusion of a lambda-secreting clone of MOPC 315 with the kappa-secreting cell line were also isolated and shown to synthesize and secrete both kappa and lambda chains. These results suggest that the nonsecretion of lambda chains was not due to a defect the secretion mechanism of the variant cells. A more likely alternative is that the lambda chains in the variant cell lines were structurally altered to a form which could not be secreted.


Contribution of vision to muscle responses in monkey during free-fall: visual stabilization decreases vestibular dependent responses. In a previous study the muscle responses from the lower limbs were studied in the Baboon (Papio-Papio) during sudden falls. On the other hand, recent findings concerning the role of vision in the control of posture during rapid perturbations in man have stimulated the present investigation. EMG activities were recorded from the fully conscious animal using chronic electrodes implanted in various muscles (splenius, quadriceps femoris, soleus, and tibialis anterior). For testing, the monkey was seated in a special chair suspended from an electromagnet and unexpectedly dropped 90 cm. EMG responses were recorded in three randomly presented conditions: with normal motion of visual world (N), with visual world stabilized with respect to the head (S) using a box covered inside by a black and white checkboard pattern surrounding the animal's head, and in total darkness (D). Results showed that condition S is accompanied in all tested muscles by a significant decrease in the EMG response; this effect is particularly evident in the interval 60--120 ms but may occur earlier. Condition D is in most of the cases accompanied by a less important decrease which is situated between condition N and condition S. These results are in contrast to previous concludions of other authors which negated the role of vision in similar situations.


Long-term effects of postnatal undernutrition and maternal malnutrition on mouse cerebral cortex. I. Cellular densities, cortical volume and total numbers of cells. Quantitative analysis of cellular densities as well as an estimate of the cortical volume and of its total cell population were performed on the cortex of postnatally undernourished mice (2--21 days) and on that of pups from malnourished mothers (gestation and lactation). Animals were followed until 180 days of age after more than 5 months of nutritional rehabilitation, and data were obtained at 10, 30, 60, and 180 days of age. The neuronal density was much higher in all cortical layers of the two experimental series, suggesting a delay in cortical maturation. Moreover, layers II, III, and IV were more delayed than layers I, V, and VI. Postnatal undernutrition had more severe effects than maternal malnutrition and the degree of recovery after a long rehabilitation was much less. The increase of the cortical volume was greatly reduced in postnatal undernutrition and to a lesser degree in maternal malnutrition. The total number of glial cells was also reduced more in postnatal undernutrition than in maternal malnutrition, but the total number of neurons was never smaller than in the controls.


Long-term effects of postnatal undernutrition and maternal malnutrition on mouse cerebral cortex. II. Evolution of dendritic branchings and spines in the visual region. Quantitative analysis of the dendritic branchings of pyramidal cells from layers V and III, as well as of the number of spines on their apical dendrites, were performed on the visual cortex of postnatally undernourished mice (2nd--21st day) and on that of pups from malnourished mothers (gestation and lactation). Animals were followed till 180 days of age after more than 5 months of nutritional rehabilitation, and data were obtained at 10, 15, 21, 30, 60, and 180 days of age. The increase and maturation of dendritic branchings and spines were much more reduced in postnatal undernutrition than in maternal malnutrition. Furthermore, permanent damage still remained at 180 days in postnatal undernutrition while almost no damage was statistically detectable after maternal malnutrition.


The velocity response of vestibular nucleus neurons during vestibular, visual, and combined angular acceleration. In alert Rhesus monkeys neuronal activity in the vestibular nuclei was measured during horizontal angular acceleration in darkness, acceleration of an optokinetic stimulus, and combined visual-vestibular stimulation. The working ranges for visual input velocity and acceleration extend up to 60 degrees/s and 5 degrees/s2. The corresponding working range for vestibular input acceleration is wider and time-dependent. During combined stimulation, that is acceleration of the monkey in the light, a linear relation between neuronal activity and velocity could be established for all neurons. Type I vestibular plus eye movement neurons displayed the greatest sensitivity and had a small linear range of operation. Other vestibular neurons were less sensitive but had a larger range of linear response to different values of acceleration. Accelerating the animal and visual surround, simultaneously but in opposite directions, results in neuronal activity proportional to relative velocity over a limited range.


Xanthumin and 8-epi-xanthatin as insect development inhibitors from Xanthium canadense Mill. Two insect development inhibitors against Drosophila melanogaster have been isolated from the leaves of X. canadense and identified as xanthumin and 8-epi-xanthatin.


Automatic recognition of squirrel monkey vocalisations by means of a filterbank. Up to now the problem of classifying squirrel monkey vocalizations has not been solved satisfactorily. The problems is now approached by a method, which consists of 2 phases. At first the monkey vocalizations are compared with the aid of physical criteria and classified by types. According to the classification of types, the membership of the individual vocalizations is tested.


Immunochemical identification of vitellogenin in the serum of the newt Triturus cristatus. The hematic yold precursor--vitellogenin--has been identified immunochemically in the serum of estrogenized females of the nest Triturus cristatus by employing an antiserum prepared against yold proteins.


Effect of dihydroergocristine infusion on tolbutamide-induced insulin secretion in man. The insulinemic response to 1 g of tolbutamide i.v. is greatly enhanced (+ 145%) after a 60-min infusion of the alpha-lytic dihydrogenated ergot alkaloid, dihydroergocristine (83.3 micrograms/min, corresponding to a total dose of 5 mg) in 7 healthy subjects. No differences are observed in the glycemic responses.


[Effect of nitroglycerin and sodium nitrite on myocardial mitochondrial respiration and oxidative phosphorylation normally and in coronarogenic ischemia]. The effect of nitroglycerin and sodium nitrite (2.10(-6), 2.10(-5) and 2.10(-4) g/ml) on respiration and oxidative phosphorylation of mitochondria of the normal and ischemized dog myocardium has been investigated in the presence of glutamate, succinate and alfaketoglutarate. The drugs inhibited ADP-activated oxygen consumption and ADP phosphorylation rate upon oxidation of succinic acid formed in mitochondria from exogenic glutamate, without changes in the phosphorylation coefficient and free oxidation rate (i. e. in the absence of ADP) in mitochondria of the healthy and ischemic heart. In the ischemic area one can also observe an inhibition of ADP-activated oxidation of endogenic succinate formed from exogenic glutamate.


[Fistula-free method of evaluating cholagogue agents on rats]. A new fistula-free method for standardizing and studying the choleretic activity of drugs on white rats is proposed. In rats under ether-urethane anestheia, a "biological test tube" of varied length where the bile is excreted, is created by ligation of the duodenum above and below the site of communication with the bile duct. In order to recover the patency of the digestive tract, the stomach or the proximal segment of the intestine is connected to its distal strip by a polyethylene tube. Supply of pancreatic juice to the "biological test tube" is eliminated by ligation of the pancreatic gland duct. The amount of bile (mg) excreted during a certain period of time, is established from the weight difference between the filled and bile-free dissected isolated segment of the duodenum. The choleretic activity of the drug is assessed in a similar way. The trial of the method performed on 50 rats, with the use of choleretic test-preparations (insulin and mebethisol) confirmed its sensitivty. The method proposed can be recommended for preliminary screening and dynamic study of the choleretic activity of drugs.


Pituitary function testing in amenorrhea-galactorrhea-hyperprolactinemia. Fifteen patients, age 16 to 55, presented with amenorrhea-galactorrhea-hyperprolactinemia. Pituitary function was evaluated by bolus injections of insulin, luteinizing hormone-releasing hormone (LHRH), and thyrotropin-releasing hormone (TRH) in 13 and by LHRH and TRH in 2. Responses to growth hormone (GH), thyroid-stimulating hormone (TSH), cortisol (F), luteinizing hormone (LH), follicle-stimulating hormone (FSH), and prolactin were measured. GH, TSH, and F responses were normal in most cases. LH responses were decreased (P less than 0.025) in patients with abnormal sellar tomography, whereas FSH responses tended to decrease with elevated prolactin levels. Prolactin responses were absent in five of the seven cases which could be evaluated. The clinical value of such testing appears to be limited to an individualized basis, although some prognosis of ovulatory response to bromocriptine therapy may be obtained from the gonadotropin response.


Acute decreases in serum prolactin concentrations caused by delta 9-tetrahydrocannabinol in nonhuman primates. The acute effects of single injections of delta 9-tetrahydrocannabinol (THC) on serum prolactin (PRL) concentrations were studied in oophorectomized female and intact adult male rhesus monkeys. Some animals were challenged with thyrotropin-releasing hormone (TRH) to determine whether THC influences pituitary responsiveness to releasing hormone. THC markedly suppresses serum PRL concentrations in animals of both sexes. TRH injections resulted in normal PRL release when administered simultaneously with or 30 minutes after THC administration. These results suggest that the serum prolactin-lowering effect of THC occurs principally at a suprapituitary level.


Effects of delta 9-tetraphydrocannabinol administration on gonadal steroidogenic activity in vivo. The effects of delta 9-tetrahydrocannabinol (THC) administration on gonadal steroidogenic activity in vivo were studied in two animal models: (1) luteal function of the rhesus monkey and (2) periovulatory steroid concentrations of rabbits treated with human chorionic gonadotropin (hCG). THC-treated monkeys demonstrated progesterone concentrations and luteal phase lengths which were not different from those in untreated controls. THC did not impair the normal pattern of response of the corpus luteum to increasing doses of hCG as measured by serum progesterone concentrations and luteal phase lengths. Rabbits treated with THC manifested normal periovulatory steroidal patterns (progesterone, testosterone, and dihydrotestosterone). These data suggest that THC does not significantly inhibit steroidogenesis by a direct action on the ovary. Its effect may be mediated by a central mechanism of action.


Immune response after vaginal application of antigens in the rhesus monkey. The immune response after  vaginal application of antigens was investigated in six sexually mature female rhesus monkeys. Two model antigens, i.e., lipopolysaccharide (LPS) OF Salmonella typhosa and abortive type T-4 coliphages were applied with or without adjuvant. A plastic sponge used as the antigen carrier was introduced into the upper vagina and placed against the ectocervix. For primary immunization, each monkey received 18 vaginal antigen applications and 10 applications for each booster course. For comparison, three other female rhesus monkeys were immunized systemically. Alum or LPS was used as adjuvant. Blood was obtained two times and cervical mucus three times weekly from each monkey. Antibodies were only barely detectable in cervical mucus after the primary vaginal immunization. However, booster treatments resulted in definite antibody responses. Specific antibodies were also detected in the circulating blood after vaginal booster immunization. The antibody level in cervical secretion in three of four cases was higher than that in circulatin blood. Systemic immunization resulted in high levels of circulating antibodies, but less than 10% appeared in cervical secretions. A characteristic decrease in antibody levels in cervical mucus was usually observed at midcycle after local immunization as well as after systemic immunization. More than 90% of T-4 coliphages applied vaginally were absorbed within 48 hours. Although alum appeared to retard the absorption of antigens, it seemed to enhance the local response. More than 90% of the antibodies to the T-4 coliphages could be removed from the serum and cervial mucus by treatment with anti-immunoglobuin G antiserum. The lymphocyte response to antigens was studied by measuring the 3H-thymidine uptake by peripheral blood lymphocytes in culture. A positive response was observed in three of three systemically immunized and in only two of six locally immunized aminals. In general, the immune response was significantly weaker after local vaginal immunization than after systemic immunization.


[Changes of plasma levels of TRH and its target hormones by two hour constant intravenous infusion of TRH tartrate in man (author's transl)]. Constant iv infusion of TRH tartrate for 2 hours was administered to normal men in a dosage of 0.5 (n=4), 1.0 (n=2) and 2 (n=4) mg/120 minutes. Measurements at every 15 minutes were performed for plasma levels of TRH, TSH, Thyroxine (T4) and Triiodothyronine (T3) by radioimmunoassay. Plasma levels of TRH increased promptly and stayed at the same levels until the end of the infusion. The Mean Clearance Rate (MRC), Half-life and Volume of Distribution of TRH were respectively, 4.62 +/- 0.53 L/min. (M +/- SE), 17.8 +/- 3.8 minutes and 112 +/- 15 L in the 0.5 mg administered group and 6.38 +/- 2.50 L/min., 9.0 +/- 1.4 minutes and 82 +/- 30 L in the 2 mg administered group. Plasma levels of TRH increased in two phases, and increments of plasma TSH were dose dependable to the dosage of TRH. Plasma levels of T4 increased gradually in the course of TRH infusion and stayed at high levels even in the withdrawal phase of TRH. Plasma levels of T3 increased markedly during and after the TRH infusion in the 0.5 mg administered group, while increments of plasma T3 were minute in the 2 mg administered group. From the above data, it is suggested that the amount of TRH production in man, which is much more than has previously been reported, may indicate the existence of an extrahypothalamic synthesis of TRH in man.


The effect of glibenclamide treatment on the insulin and glucagon responses to oral glucose and galactose in maturity onset diabetics. Glibenclamide has been shown to stimulate an insulin releasing factor in the duodenum. The possibility that this effect is of importance in its hypoglycaemic action was investigated by studying the effect of galactose on insulin release before and after treatment with glibenclamide; galactose stimulates insulin release when given orally but has no effect when given parenterally; thus its ability to release insulin appears to reside in an action on a gut factor. Measurements of plasma glucose, insulin and glucagon were made on twelve maturity onset diabetic patients following an oral glucose tolerance test and an oral galactose tolerance test before and after one week of treatment with glibenclamide. Glibenclamide significantly reduced the blood glucose levels. Both basal insulin and basal glucagon levels were unchanged. The insulin response to oral glucose was enhanced. Glucagon levels before treatment did not suppression of glucagon levels. Galactose stimulated insulin release but insulin levels before and after treatment were identical. An effect of glibenclamide on gut insulin releasing activity was not demonstrated but the galactose tolerance test provides a useful technique by which to examine the enteroinsular axis.


Global and effective synergism of amikacin, gentamicin or tobramycin when combined with carbenicillin against Pseudomonas aeruginosa. The authors describe a study to evaluate the synergistic effects of the combinations of amikacin-carbenicillin, gentamicin-carbenicillin and tobramycin-carbenicillin at the associated minimal bactericidal concentrations and at doses related to the serum levels reached by the drugs in the blood.


The role of mesna in the treatment of status asthmaticus in children. Status asthmaticus in children is a life-threatening condition that calls for a comprehensive set of therapeutic measures, viz. evacuation of tracheobronchial secretions, parenteral drug administration and improvement of alveolar gas exchange. In the authors' experience, that therapeutic procedure can be much improved by bronchial lavage with mesna. This is illustrated by the good results obtained in fourteen patients with status asthmaticus and in another forty-four cases of global respiratory insufficiency. Evaluation was by blood gas values and X-ray pictures. The method used for bronchial lavage is described.


Chronic liver disease after acute non-A, non-B viral hepatitis. We have analyzed the frequency of chronicity and its distribution according to epidemiologic background following acute non-A, non-B hepatitis. Eighteen of 45 cases (40%) developed chronic liver disease. The incidence of chronicity was significantly higher following transfusion and among drug addicts (54% and 58%) than among patients without obvious source of infection (20%). Chronic active hepatitis developed in 4 of 13 patients (31%) with posttransfusion hepatitis. This lesion was not observed among the addicts or the patients without obvious source for the acute hepatitis.


The genetics of a small autosomal region of Drosophila melanogaster containing the structural gene for alcohol dehydrogenase. I. Characterization of deficiencies and mapping of ADH and visible mutations. The position of the structural gene coding for alcohol dehydrogenase (ADH) in Drosophila melanogaster has been shown to be within polytene chromosome bands 35B1 and 35B3, most probably within 35B2. The genetic and cytological properties of twelve deficiencies in polytene chromosome region 34--35 have been characterized, eleven of which include Adh. Also mapped cytogenetically are seven other recessive visible mutant loci. Flies heterozygous for overlapping deficiencies that include both the Adh locus and that for the outspread mutant (osp: a recessive wing phenotype) are homozygous viable and show a complete ADH negative phenotype and strong osp phenotype. These deficiencies probably include two polytene chromosome bands, 35B2 and 35B3.


The genetics of a small autosomal region of Drosophila melanogaster containing the structural gene for alcohol dehydrogenase. II. Lethal mutations in the region. Forty-seven lethal mutations and alleles of nine visible loci (including alcohol dehydrogenase) have been mapped by both deficiency mapping and, in most cases, by recombination mapping to a small region (34D-35C) of chromosome arm 2L of Drosophila melanogaster. The lethals fall into approximately 21 complementation groups, and we estimate that the total number of lethal plus visible complementation groups within the 34-band deficiency, Df(2L)64j, is approximately 34, a remarkable numerical coincidence. The possible genetic significance of this coincidence is discussed. Lethals mapping close to the structural gene for alcohol dehydrogenase, both distally and proximally, have been identified and will be used for the construction of selective crosses for the study of exchange within this locus. Despite many abnormal cytological features (e.g., ectopic pairing, weak points) region 35 of chromosome arm 2L does not display any unusual genetic features; indeed, in terms of the amount of recombination per band and the average map distance between adjacent loci, this region is similar to that between zeste and white on the X chromosome.


Hybrid dysgenesis in Drosophila melanogaster: the biology of female and male sterility. High levels of female and male sterility were observed among the hybrids from one of the two reciprocal crosses between a wild strain of D. melanogaster known as pi2 and laboratory strains. The sterility, which is part of a common syndrome called hybrid dysgenesis, was found to be associated with the rudimentary condition of one or both of the ovaries or testes. All other tissues, including those of the reproductive system were normal, as were longevity and mating behavior. The morphological details of the sterility closely mimic the agametic condition occurring when germ cells are destroyed by irradiation or by the maternal-effect mutation, grandchildless. We suggest that sterility in hybrid dysgenesis is also caused by failure in the early development of germ cells. There is a thermo-sensitive period beginning at approximately the time of initiation of mitosis among primordial germ cells a few hours before the egg hatches and ending during the early larval stages. Our results suggest that hybrid dysgenesis, which also includes male recombination, mutation and other traits, may be limited to the germ line, and that each of the primordial germ cells develops, or fails to develop, independently of the others. This hypothesis is consistent with the observed frequencies of unilateral and bilateral sterility, with the shape of the thermosensitivity curves and with the fact that males are less often sterile than females. The features of this intraspecific hybrid sterility are found to resemble those seen in some interspecific Drosophila hybrids, especially those from the cross D. melanogaster X D. simulans.


Aberrations induced in chromosomes of somatic cells of Drosophila melanogaster irradiated in C-metaphase. Experiments conducted on the X irradiation of neural ganglia of Drosophila melanogaster are described. The ganglia were placed in saline containing colchicine. After two hours, they were irradiated and then samples were fixed at 5,15,25,35 minutes from the beginning of irradiation. The results obtained show that the aberration level increases with time subsequent to fixing. This increase takes place first for chromatid deletions and then for isochromatid deletions and chromatid exchanges. Gaps and subchromatid exchanges do not, on the contrary, show any increase with time. We did not observe a difference in radiosensitivity between the sexes. Some hypotheses are put forth in an attempt to explain these results.


Lysosomal dysfunctions associated with mutations at mouse pigment genes. Melanosomes and lysosomes share several structural and biosynthetic properties. Therefore, a large number of mouse pigment mutants were tested to determine whether genes affecting melanosome structure of function might also affect the lysosome. Among 31 mouse pigment mutants, six had 1.5- to 2.5-fold increased concentrations of kidney beta-glucuronidase. Three mutants, pale ear, pearl and pallid, had a generalized effect on lysosomal enzymes since there were coordinate increases in kidney beta-galactosidase and alpha-mannosidase. The effects of these three mutations are lysosome specific since rates of kidney protein synthesis and activities of three nonlysosomal kidney enzymes were normal. Also, the mutants are relatively tissue specific in that all had normal liver lysosomal enzyme concentrations.--A common dysfunction in all three mutants was a lowered rate of lysosomal enzyme secretion from kidney into urine. While normal C57BL/6J mice daily secreted 27 to 30% of total kidney beta-glucuronidase and beta-galactosidase, secretion of these two enzymes was coordinately depressed to 1 to 2%, 8 to 9% and 4 to 5% of total kidney enzyme in the pale-ear, pearl and pallid mutants, respectively. Although depressed lysosomal enzyme secretion is the major pigment mutant alteration, the higher lysomal enzyme concentrations in pearl and pallid may be partly due to an increase in lysosomal enzyme synthesis. In these mutants kidney glucuronidase synthetic rate was increased 1.4- to 1.5-fold.--These results suggest that there are several critical genes in mammals that control the biogenesis, processing and/or function of related classes of subcellular organelles. The mechanism of action of these genes is amenable to further analysis since they have been incorporated into congenic inbred strains of mice.


[Identification of the restriction and modification systems in Streptomyces strains]. Host range of actinophage phi C31, VP5 and Pg81 in respect to 109 strains of Streptomyces genus and hybrid strain Rcg2 from the cross S. coelicolor A3(2)XS. griseus Kr was studied. The existence of RM-systems in strains S. griseus Kr15, S. griseus Kr20, Rcg2, S. griseofovillus 43 was shown using phage Pg81. Mutants of Pg81 were observed which to some extent lost snesitivity to RM-system in the strain Rcg2. The presence of RM-system in S. lividans 67 was demonstrated by the phage VP5.


[Ecologically conditioned changes in gene frequencies for polymorphic systems in the nonnative population of the northeastern USSR]. Examination of 1051 healthy residents of Magadan in terms of nine polymorphic systems has shown changes in phenotype frequencies depending on duration of the persons' staying in the North. The frequency pattern of phenotypes is significantly different among men and women. The phenotype frequencies of alkaline serum phosphatase and acidic erythrocyte phosphatase were found in a more strict correlation with the northern residence term. The part played by selective migratory behaviour in development of genetic structure of population is discussed and also possible ecologic factors resulting in phenotype and gene frequency changes are analysed.


[Thiophosphamide induction of sister chromatid exchanges at various phases in the cell cycle of a Chinese hamster cell culture]. Influence of three concentrations of thiophosphamide (thioTEPA) on the formation of sister chromatid exchanges (SCE) has been studied at different phases during 2 cell cycles in cultured Chinese hamster cells. It is shown that the frequency of SCE does not differ from the control level under the effect of the mutagen on cells in the G2 phase of the first cell cycle from the moment of harvesting. Thiophosphamide induces the same number of SCE at S, G1 stages of the first cell cycle and G2 of the second one till the moment of harvesting. The number of SCE correlates in a direct proportion with a concentration of thiophosphamide. A scheme of forming SCE is proposed.


Restriction-fragment map of the temperate Bacillus subtilis bacteriophage SPO2. The endonucleases BglI, BglII, EcoRI, SalI, SmaI, and XbaI were used to fragment the phage SPO2 DNA. Electrophoretic analysis using ethidiumbromide agarose gels showed the phage to have nine BglI sites, one BglII site, four EcoRI sites, one SalI site, one SmaI site, and six XbaI sites. Using partial digestions, multiple endonuclease digestion, and autoradiography the fragments were sized and ordered into a circular map of 23 Md. Such an analysis locates the endonuclease sites, indicates which endonucleases are potentially useful in cloning with SPO2, and allows insertions and/or deletions in the SPO2 DNA to be characterized.


Isolation and characterization of viable deletion mutants of Bacillus subtilis bacteriophage SPO2. Spontaneous deletion mutants of the bacteriophage SPO2, which are viable and retain their temperate character, were isolated using a heat-EDTA enrichment step. They were identified by endonuclease digestion and agarose-gel electrophoresis of phage DNA. Two of the nine mutants were characterized in detail. Both mutants have a 2.3 Md deletion removing the single BglII site and two of the XbaI fragments. The deletion extends 1.0 Md to one side of the former BglII site and 1.3 Md on the other side. This region of the SPO2 genome is non-essential for either lysogeny or viable phage production and thus is a suitable region for the insertion of exogenous DNA fragments.


Correlation of activation and aggregation of platelets. Discrimination between anti-activating and anti-aggregating agents. Shape change and release reaction indicate different degrees of the complex platelet response termed activation. Aggregation is a variable consequence of activation. Aggregation shows a temperature dependency opposite to that shown by the preceding shape change. Aggregation increases at lower temperature and requires, in contrast to activation, extracellular Ca2+, stirring, and at a low degree of activation the presence of fibrinogen. Aggregation can enhance activation by triggering prostaglandin endoperoxide-thromboxane synthesis. If activation reaches a high level associated with the release reaction, activation is further amplified by prostaglandin endoperoxide-thromboxane synthesis emerging independent of aggregation. These mechanisms of amplification of activation are blocked by indomethacin, an inhibitor of prostaglandin endoperoxide-thromboxane synthesis=anti-activating agent. In contrast, anti-aggregating agents, exemplified here with n-acetyl neuraminic acid, attack the aggregation of activated platelets but neither activation nor prostaglandin endoperoxide-thromboxane synthesis. Its anti-aggregating effect, in addition, enables n-acetyl neuraminic acid to imitate the inhibitory effect of indomethacin on the feedback amplification which results from aggregation. Anti-aggregating agents as characterized here may open a new valuable concept for anti-aggregation in vivo.


Effect of estrogen on postovulatory LH surge in baboons. In normally cycling female baboons, an LH surge appeared prior to ovulation, in addition, another LH surge (postovulatory LH surge) was observed within two days after ovulation. An attempt was then made to determine the effect of postovulatory LH on the luteinization of corpus luteum in baboons. Injections of 300 micrograms estradiol benzoate were given at 09.00 and 16.00 hr daily for 5 days following ovulation; the plasma level of LH was increased, but plasma progestin was suppressed. These results infer that the injected estrogen (estradiol benzoate) may inhibit the luteotrophic effect of postovulatory LH on the corpus luteum, therefore, plasma progestin remains lower even though postovulatory LH is elevated.


Subcellular distribution of aromatase in human placenta and ovary. The aromatization of androstenedione in human ovarian microsomes is inhibited by an antibody to porcine hepatic microsomal NADPH-cytochrome c reductase. Likewise, the antibody inhibits aromatization in mitochondria isolated from human ovaries and placentae. A given quantity of the antibody produces the same percent inhibition of aromatization in microsomes and mitochondria of both ovaries and placentae. These data, in addition to the low specific activity observed for the mitochondrial aromatase, indicate that aromatization in mitochondria probably results from microsomal contamination.


Parenteral nutrition: first decade, trends. Parenteral feeding has saved lives and enhanced therapy of patients who cannot eat. Future trends of this technique for patients, physicians, hospitals are discussed.


Differences in the genetic control of primary and secondary antibody responses. Primary and secondary antibody responses to f and s antigens of Salmonella typhimurium have been studied in H and L lines of mice genetically selected for primary reponse to sheep erythrocytes (SE) (Selection I). The range of interline separation obtained (non-specific effect of Selection I) was as large as for the selection antigen in the primary response to f antigen and slightly smaller in the primary response to s antigen. For these two antigens the interline difference was reduced after booster. The kinetics of responses were compared with those obtained in H and L lines of Selections III and IV carried out for secondary responses to f and s antigens of S. typhimurium respectively (specific effect of Selection III and IV). The genetic analysis was made in Selection I from the variances of individual agglutinin titres obtained in large groups of interline hybrids immunized with S. typhimurium. These calculations gave a reliable estimate of the effective number of independent loci regulating primary and secondary responses. The results demonstrated a major difference in the genetic control: a single locus regulated the secondary response to f antigen while six loci were involved in the control of the primary response. A similar difference was evident for s antigen. The primary response was likely to be under polygenic regulation although the effective number of loci could not be calculated, while the secondary response was under monogenic control.


A biclonal origin of two monoclonal proteins, IgG3(kappa) and IgA1(lambda), from a single patient. The cellulose acetate electrophoretic pattern of the serum from patient Sik disclosed two distinct peaks, representing two monoclonal proteins. On immunoelectrophoresis the two M-components were found to differ in heavy chain class as well as in light chain type, IgG3(kappa) and IgA1(lambda). Serum immunoglobulin levels remained relatively constant over a period of 7 years and no clinical symptoms of a malignant deterioration occurred. It was found that the isolated M-components did not share idiotypic antigenicity. Bone marrow cells synthesizing the monoclonal proteins were identified by means of the immunofluorescent technique using isotypic as well as idiotypic antisera. Two distinct monoclonal cell populations were observed, containing either the IgG3(kappa) or the IgA1(lambda) monoclonal protein. The alpha 1-chain belonged to the VHIII subgroup, whereas the gamma 3-chain was found to be blocked. Subsequent sequence determination showed the gamma 3-chain to belong to the VHIII subgroup. It was concluded that the two M-components in the serum of patient Sik resulted from two independent neoplastic transformations.


Proteolytic transformation of SC5b-9 into an amphiphilic macromolecule resembling the C5b-9 membrane attack complex of complement. Proteolysis of fluid-phase SC5b-9 left a major part of the macromolecule intact and caused transition of the molecule from a hydrophilic to an amphiphilic state. The transformed complex exhibited neoantigens characteristic of the C5b-9 membrane attack complex of the complement. It yielded an SDS gel electrophoresis pattern that was similar, but not identical to that of the proteolysed, membrane attack complex. The proteolytically altered SC5b-9 complex bound lipid and incorporated into artificial lipid vesicles to yield a membrane-bound structure resembling the C5b-9 complement lesion.


Evaluation of a new polyvalent Pseudomonas vaccine in respiratory infections. A new polyvalent, cell wall extract, Pseudomonas aeruginosa vaccine (PEV-01), was evaluated by using a guinea pig model of experimental Pseudomonas pneumonia. Guinea pigs routinely developed fourfold rises in serum hemagglutinating Pseudomonas antibodies after four vaccine injections given over 2 weeks. Vaccinated animals survived an intratracheal Pseudomonas challenge (1 X 10(8) colony-forming units) significantly better (13 of 14 survived) than did a control group (5 of 14 survived) (P less than 0.01). Clearance of viable Pseudomonas from lung tissue was significantly better in vaccinees than controls at both 3 h (P less than 0.02) and 6 h (P less than 0.05) after infection. Both gross and histological examinations of lung tissue revealed less pulmonary tissue damage in vaccinated animals following Pseudomonas infection. Thus, PEV-01 Pseudomonas vaccine appears capable of eliciting a specific protective response in the guinea pig respiratory tract.


Isolation of a cured strain from Corynebacterium diphtheriae PW8. A nonlysogenic, non-toxinogenic strain was isolated from the PW8 strain of C. diphtheriae by two-step ultraviolet induction. This strain was lysed by phages derived from the PW8 strain but not by beta phages from the C7(beta) strain. When this cured strain was lysogenized with phages from the PW8 strain, toxin production by the resulting lysogens was about a half or a quarter of that of the parent strain PW8.


Serotypes of beta-hemolytic Treponema hyodysenteriae. Cultures form 13 isolates of pathogenic, beta-hemolytic Treponema hyodysenteriae from 11 geographically separate outbreaks and 2 experimentally induced cases of swine dysentery were lyophilized and extracted with hot phenol-water. The resulting water phases were examined serologically with antisera produced in rabbits against whole-cell bacterins of the 13 isolates for evidence of antigenic classes within the species. Water-phase antigens gave precipitin reactions with homologous antisera. Results from cross-testing of each water phase with each antiserum showed four serologically distinct groups among the isolants examined. Based on precipitin reactions in agarose gel, four serotypes of pathogenic, beta-hemolytic T. hyodysenteriae are proposed.


Role of rabbit lysozyme in in vitro serum and plasma serum bactericidal reactions against Bacillus subtilis. The antibacterial activity of purified rabbit lysozyme was kinetically investigated at concentrations comparable to those in normal rabbit serum and plasma serum. The bactericidal capability, lysozyme content, and electrophoretic composition of "purified beta-lysin," fractionated from normal rabbit serum, were also examined. In contrast to the extensive antibacterial activity of dilute normal rabbit serum observed in vitro, rabbit lysozyme was only weakly bactericidal for Bacillus subtilis. Inhibition of lysozyme enzymatic and bactericidal activities in normal rabbit serum by antilysozyme immunoglobulin G slightly reduced the initial rate of killing. The addition of neutralizing antibody or histamine (another lysozyme inhibitor) to partially purified bactericidal serum fractions had no effect on killing kinetics. Increasing the ionic strength of reaction mixtures containing normal serum or partially purified bactericidal fractions to levels which completely inhibited lysozyme activity resulted in stimulation of their respective killing kinetics. The addition of inhibitors to normal rabbit plasma serum completely eliminated its bactericidal activity. With regard to the killing of B. subtilis by rabbit and human blood fractions, these analyses clearly demonstrated that (i) although lysozyme is not a significant antibacterial component of normal rabbit serum, it represents the principal factor in normal rabbit plasma serum; (ii) different primary bactericidal mechanisms which are not detectable by singlepoint analyses operate in the sera of different species; and (iii) purified beta-lysin isolated from normal rabbit serum by the classical procedure is a heterogenous mixture of components.


Adoptive transfer of protection against Trypanosoma cruzi with lymphocytes and macrophages. Female C57BL/6J mice were infected with Trypanosoma cruzi and subsequently given macrophages or lymphocytes from syngeneic donors which had recovered from the acute infection. Mice which received immune peritoneal macrophages, splenic lymphocytes, or lymph node lymphocytes developed lower mean parasitemias and cumulative mortalities than did recipients of nonimmune cells. Neither peritoneal lymphocytes nor splenic macrophages were protective, however. These studies indicate that splenic and lymph node lymphocytes are effective in transferring protection against T. cruzi, whereas the macrophage is somewhat less effective.


Outer membrane antigens of Neisseria meningitidis group B serotype 2 studied by crossed immunoelectrophoresis. This study shows that the capsular polysaccharide, protein, and lipopolysaccharide antigens from the outer membrane of Neisseria meningitidis group B serotype 2 may be identified by crossed immunoelectrophoresis. By using this technique, seven precipitates were resolved when outer membrane preparations were reacted against goat anti-whole cell group B type 2 antiserum. Most of these precipitates were identified by comparison with purified reference preparations. Different outer membrane preparations, reflecting different growth conditions, varied in their compositions of lipopolysaccharide, protein, and polysaccharide. Detergent treatment altered the protein and lipopolysaccharide precipitation patterns. In the presence of detergent, the lipopolysaccharide did not precipitate, and the electrophoretic migration of the protein antigens decreased. Crossed immunoelectrophoresis is a useful qualitative method for analysis of the antigenic components of the meningococcal outer membrane. The crossed immunoelectrophoresis with intermediate gel technique is presently being used to measure the human immune response to the different cell surface components.


Inhibition of the growth of Neisseria meningitidis by reduced ferritin and other iron-binding agents. Serogroups of N. meningitidis were characterized as virulent or avirulent according to their capacity to establish meningococcal infection in mice. An agar plate diffusion technique demonstrated that iron had a definite growth-supporting role for both of these meningococcal types. The avirulent strains could use ionic or chelated iron as well as the virulent strains. Iron-reversible growth inhibition occurred to the same extent for both bacterial types in the presence of the synthetic iron-chelating agents Desferal and ethylenediamine-di-orthohydroxy phenylacetic acid. A difference in response was demonstrated for these bacterial types when grown in the presence of various iron-binding proteins from animal body fluids and tissues. The growth of the avirulent strain was inhibited to a greater degree by egg white conalbumin. The humoral iron-binding protein transferrin showed a significant inhibitory capacity only when used in conjunction with bicarbonate. Under conditions of increased iron saturation of this protein, the avirulent strain was inhibited to the furthest extent. In the presence of ferritin, the cellular iron-binding protein, which had been reduced, inhibition of the growth of either strain type did not occur on iron-poor media (less than 5 micrograms/100 ml). However, with the incorporation of iron into the media, the inhibitory effect of the protein became evident. As the concentration of iron increased, the inhibition increased to a certain level and subsequently declined. A substantial difference in the ability of the avirulent type to grow in the presence of reduced horse spleen ferritin was observed. For this microorganism, a correlation appears to exist between the capacity to grow by utilizing the available iron in the presence of reduced ferritin and the ability to establish infection. The host protein ferritin, in the reduced state, apart from simply being a storage protein for iron, can prevent the growth of a procaryotic organism. Our experiments suggest a role for ferritin in the prevention of emningococcal disease. A cehmotherapeutic potential for Desferal is also implied.


Purification of a surface-specific soluble antigen from Listeria monocytogenes. A complex antigenic preparation obtained from Listeria monocytogenes serovariant 4b by freeze-pressing, centrifugation, and gel filtration treatment was studied by crossed immunoelectrophoresis, with the aim of preparing an antigenic fraction that could be used to investigate the serological response to listeric infection. Of 17 immunoprecipitates revealed in the soluble extract, one of three major antigens (designated antigen 2) was shown to be a strong antigen in humans or rabbits infected with L. monocytogenes serovariant 4b. A monospecific antiantigen 2 serum was obtained and used to prepare a serologically homogeneous antigen by immunoadsorption. Antigen 2, most probably located on the bacterial surface, is common to all serovariants of L. monocytogenes and to Listeria grayi and is not shared by the main bacterial species known to have common antigens with L. monocytogenes.


Protection against group B meningococcal disease: evaluation of serotype 2 protein vaccines in a mouse bacteremia model. A mouse bacteremia model was used to evaluate the immunogenicity and protection against challenge provided by five different meningococcal serotype 2 vaccines. Mice vaccinated with serotype 2 protein vaccines had levels of bacteremia reduced by at least 100-fold after challenge with group B serotype 2 miningococci. Mice vaccinated with serotype 2 protein vaccine and challenged with group C serotype 2 meningococci showed 10-fold or less reduction in bacteremia. Vaccines were primarily serotype specific since no increase in protection was observed after challenge with either group B serotype 4 or group C nontypable meningococci. Serotype 2 antibody levels, measured by the bactericidal assay and the enzyme-linked immunosorbent assay 3 weeks after immunization, demonstrated a graded dose-response which correlated with protection up to 40 weeks after vaccination with a single 10- or 25-micrograms dose of serotype 2 protein vaccine. A 1-microgram booster dose of serotype 2 vaccine, given 2 weeks after primary immunization, significantly increased both bactericidal (P less than 0.01) and enzyme-linked immunosorbent assay (P less than 0.02) titers. The data obtained from the mouse bacteremia model indicate that serotype 2 protein vaccines are stable and immunogenic, and protect mice against challenge with group B serotype 2 meningococci.


Passage of Pseudomonas aeruginosa in compromised mice. There was no appreciable increase in the virulence of Pseudomonas aeruginosa PAO-1 after six passes through infections in mice. When strain PAO-1 was passed through mice compromised with iron and methotrexate, the virulence of the passed bacteria increased for normal as well as compromised mice. Bacteria harvested from intraperitoneal passage in compromised mice were more virulent than bacteria harvested from intrathoracic passage. These bacteria expressed 50% lethal dose values distinctive of the respective bacterial isolate when injected intraperitoneally, intrathoracically, or intravenously. Differences between bacteria from intraperitoneal and intrathoracic passage were apparently due to differences in the selective pressures in the two sites of infection, because the intrathoracically passed bacteria assumed the virulence characteristics of the intraperitoneally passed bacteria after intraperitoneal passage in compromised mice.


Gamma-irradiated scrub typhus immunogens: broad-spectrum immunity with combinations of rickettsial strains. Scrub typhus immunogens were prepared from Rickettsia tsutsugamushi strains Karp, Kato, Gilliam, Kostival, and Buie by exposing frozen infected yolk sac suspensions to 300 krad of gamma radiation. Mouse protection tests showed that each of the irradiated immunogens protected C3H/HeDub mice against high challenge levels of Karp and Gilliam, but that none of these single-strain immunogens were capable of protecting against all five of the challenge strains. Broad-spectrum protection was achieved by using combinations of three strains of irradiated rickettsiae in a vaccination regimen of three injections at 5-day intervals. A comparison of vaccination efficacy employing three such combinations (Karp-Gilliam-Kato, Karp-Kostival-Kato, and Buie-Kostival-Kato) indicated that both sequential administration of strains on successive vaccination days and multiple injections of trivalent mixtures produced protective responses superior to those obtained with single-strain immunogens. Trivalent mixtures of rickettsiae exhibited a striking synergistic effect on the immune response of C3H/HeDub mice and elicited a protective response against Kato challenge that could not be obtained with any single-strain immunogen. Mice vaccinated with the trivalent Karp-Gilliam-Kato immunogen resisted challenge with more than 10(3) 50% mouse lethal doses of Karp and Gilliam for 12 months, and were resistant to similar levels of challenge with Kato and Buie for 6 months.


Enzyme-linked immunosorbent assay for bovine immunoglobulin subclass-specific response to Brucella abortus lipopolysaccharides. An enzyme-linked immunosorbent assay was developed to follow the bovine response, by immunoglobulin class and subclass, to defined smooth and rough lipopolysaccharides (LPS) of Brucella abortus. Binding to smooth LPS of immunoglobulin G1 (IgG1) and IgG2 in sera from Brucella-infected animals was significantly greater than binding in sera from normal uninfected animals. Competition or steric blocking among IgM, IgG1, and IgG2 for binding sites on smooth LPS was shown to occur. Binding of IgM to Brucella smooth LPS with sera from uninfected animals was elevated above the assay control levels, and attempts to eliminate this nonspecific IgM binding were not successful. The same levels of nonspecific IgM binding were also seen with Brucella rough LPS, Escherichia coli LPS, and Pseudomonas solanacearum LPS. Sera from some, but not all, Brucella-infected animals showed elevated binding of IgG1 and IgM to both E. coli LPS and Brucella rough LPS as well as to Brucella smooth LPS. This was interpreted as specific antibody. Cross-reactions between B. abortus smooth or rough LPS and E. coli LPS could not be shown by immunodiffusion.


Use of glutaraldehyde-modified timothy grass pollen extract in nasal hyposensitisation treatment of hay fever. 12 patients suffering from grass pollen hay fever were treated for 14 weeks pre- and co-seasonally by intranasal self-administration of an aqueous solution of a glutaraldehyde-treated timothy grass pollen allergen. These patients had a statistically significant decrease in nasal symptom scores during the grass pollen peak period and in nasal challenge end-point titre after the season compared to placebo-treated patients. No significant effect was seen on the eye symptoms. 1 patient withdrew from the trial as a consequence of too strong local nasal reactions during treatment. Most other patients treated with active material reported mild local reactions during the first minutes after administration of the nasal spray. In the actively treated group a significant increase in serum and nasal secretion of grass pollen specific IgE, IgG and IgA antibodies was obtained during the treatment. In contrast, in the placebo group a significant increase in IgE antibody levels in serum and secretion occurred during the pollen season. The reduction in symptoms and increase in antibody production together with the simplicity of the procedure makes this approach to immunotherapy attractive.


Canadian survey to determine the rate of drug resistance to isoniazid, PAS and streptomycin in newly detected untreated tuberculosis patients and retreatment cases. In 1975, a survey was carried out in Canada to determine the primary and acquired drug resistance of M. tuberculosis isolates to isoniazid (INH), para-aminosalicylic acid (PAS) and streptomycin. The results of this investigation were compared with those of the primary drug resistant study of Armstrong, undertaken in 1963-64. It revealed that primary drug resistance has increased from 4.9% to 6.3%. The increase is mainly due to immigrants having arrived in this country during the last 12 years. In these newcomers the primary resistance rate was 11.5%. Moreover, 57.8% of the immigrants examined in the survey were of Asian origin, with a drug resistance rate of 11.7%, while 15.6% had arrived from South Europe with a resistant ratio of 16.7%. In retreatment cases, the national average of drug resistance was 26.4%. Among the Canadian provinces, the highest drug resistance rate in retreatment patients (40%) was found in Quebec. While in primary resistance Streptomycin exhibited the highest incidence, in retreatment cases isoniazid resistance proved to be more frequent. In natives, the rates and patterns of primary and acquired resistance were very similar to those observed in other Canadian born patients.


Repairs of X-ray induced sublethal damage in class B oocytes of Drosophila melanogaster. Repair of X-ray-induced sublethal damage (Elkind-type recovery) in class B oocytes of Drosophila melanogaster was studied. Newly hatched females of two different stocks were treated with either acute or fractionated exposures. For the fractionation experiments a constant time interval of one hour between the dose fractions was used. As genetic endpoints dominant lethality, chromosome aberrations (detachments) and non-disjunction were studied. The repair of X-ray-induced sublethal damage in class B oocytes is expressed as a reappearance on the initial shoulder in the fractionation curve. For dominant lethality it could be shown that less sublethal damage is repaired in oocytes of Berlin wild females than in those of attached-X females (on the average 76 per cent and 101 per cent respectively). Complete repair (about 100 per cent) was observed for detachments in occytes of attached-X females. Within the dose ranges used no radiation effects on non-disjunction could be observed. The results are interpreted to show that in class B oocytes (1) sublethal damage is due to chromosome breaks and/or lesions leading to breaks and (2) X-ray-induced dominant lethality is the consequence of chromosome damage (true dominant lethals).


Time dependence of Elkind-type recovery in class B oocytes of Drosophila melanogaster. Recovery from X-ray-induced damage in class B oocytes of Drosophila melanogaster was studied by the dose-fractionation technique. A total dose of 500 R was delivered either as a single exposure or as two fractions of 2000 R and 3000 R separated by increasing time intervals. The use of attached-X females made it possible to study simultaneously the induction of dominant lethals and of chromosome aberrations (detachments of the attached-X chromosome). The same repair kinetics were observed for sublethal damage and for the lesions leading to detachments. The time-response curves are of similar shape: a plateau is reached within 20 to 30 min and half of the repairable damage disappears in 5 to 7 min. It is concluded that the same type of X-ray-induced primary lesion in chromosomes is responsible for the induction of detachments and for dominant lethals. As primary lesions actual chromosome breaks or lesions leading to breaks and chromosome rearrangements are assumed.


The nature of the damage to Escherichia coli DNA induced by gamma-irradiation. Quantitative studies of the number of gamma-induced single-strand breaks (SSBs) and enzyme-labile sites (ELSs) were performed using the model of Col E1 plasmids, which undergo transition from the covalently closed form (CCF) into the open circular form (OCF) during gamma-irradiation of the plasmid-bearing strain E. coli JC 411. By adding 0.5 MEDTA the repair endonucleases of the cell, which effect the transition of ELSs into SSBs during and after gamma-irradiation, were totally inhibited. It was found thless than 15 per cent of the number of gamma-induced lesions are primarily induced SSBs. About the saditions of direct radiation damage. The conclusions are that (1) the contribution of the direct radiation effect in the cell is greater than that of the indirect effect; (2) the main type of gamma-induced lesions are the ELSs (most of which--more than 75 per cent--are alkali-stable; (3) the enzymatic incision of gamma-induced ELSs into SSBs is effected very quickly, mainly during irradiation; and (4) 0.5 MEDTA is a universal inhibitor of repair processes in cell, including the action of N-glycosidases and endonucleases.


Patterns of distribution of phosphomono-esterases on surfaces of demineralized bone. Decalcification over short periods (5 days) with MnNa2 EDTA, MgNa2 EDTA and EGTA according to a method described in the present paper, creates sections of high quality with simultaneous good preservation of phosphomonoesterases on bone surfaces. In fact, the enzyme distribution seems to be comparable to that obtained by using undecalcified sections. Na2 EDTA creates, on the other hand, poor preservation of alkaline phosphatase probably due to the fact that this chelate contrary to the other chelates removes the essential metal from the protein, leaving an unstable enzyme molecule which undergoes denaturation. Decalcification over longer periods (15 days) does not influence the pattern of distribution of acid phosphatase, whereas the alkaline phosphatase reaction becomes depressed in certain surface areas. The significance of this differential distribution is discussed. It might be an indication of differential processes of bone transformations in such a way that bone surfaces corresponding to areas of enzyme reactions are depository whereas bone surfaces corresponding to areas of lack of enzyme reaction are resorptive. New experimental designs are, however, necessary before the phenomenon is fully perceived. Two different coupling agents were used in connexion with the demonstration of acid phosphatase reaction. When HPR was used as the coupler the final enzyme distribution coincided with that usually described in the literature, i.e., strong reaction of cells adjacent to resorptive surfaces and weak reaction of cells adjacent to depository surfaces. When, however, Fast dark blue R was used all surface cells reacted markedly. This method also revealed certain cell types with nuclear reaction.


Nutritional concepts in the treatment of head and neck malignancies. Cancer cachexia should no longer be a contraindication to adequate antineoplastic treatment. Current methods of nutritional assessment allow one to identify malnutrition and to follow the nutritional status of the patient throughout the cancer-management program. Enteral nutritional repletion and maintenance remain the ideal course of action, but the gastrointestinal tract is not always readily available or advisable for use; in such circumstances, intravenous hyperalimentation (IVH) may be indicated. The properly nourished patient better tolerates cancer therapy, experiences fewer complications of malnutrition (e.g., sepsis and poor wound healing), and has a better-functioning immune system than does his malnourished counterpart. This article reviews methods of nutritional assessment, delineates indications and techniques for nutritional repletion, and summarizes the results obtained.