The dependence of T wave alternans on diastolic resting period duration. The dependence of T-wave alternans on diastolic resting period duration as well as on regional differences in monophasic action potential duration was studied in the in situ dog heart. T-wave alternans was elicited when diastolic resting period shortened to values less than 41 +/- 6% of the cardiac cycle. This could be attained either by increasing the driving rate, or lengthening action potential duration by inducing hypocalcemia. During T-wave alternans, monophasic action potentials recorded from anterior and posterior left ventricular surface showed an alternation in duration; the degree in alternation in both regions differed and related to T-wave polarity.


Treatment of Prinzmetal's angina due to coronary artery spasm using verapamil: a report of three cases. In 3 patients with Prinzmetal's variant angina there were inferior epicardial injury patterns on the electrocardiogram, corresponding to right coronary artery narrowing. Attacks were provoked by beta-adrenergic blockade and stopped by nitroglycerine and atropine. Spasm of the right coronary artery was demonstrated in the 2 patients who had high parasympathetic activity. In all 3 cases attacks disappeared on verapamil. We suggest that calcium-antagonists, which dilate the coronary arteries, should be used in the treatment of Prinzmetal's angina pectoris rather than beta-adrenergic blockers, which do not have this property.


Effects of intravenous nitroglycerin on hemodynamics and ischemic injury in patients with acute myocardial infarction. In 24 patients with acute myocardial infarction intravenous nitroglycerin in 2 different dosages was administered and the effect on hemodynamics and extent of myocardial ischemia was investigated. According to the initial left ventricular filling pressure (LVFP) the patients were divided in Group I: LVFP below 20 mm Hg (13 patients) and in Group II: LVFP above 20 mm Hg (11 patients). Following an infusion of 3 mg of nitroglycerin in the first and 6 mg in the second hour, a significant decrease (P less than 0.0001) in the filling pressure (in Group I from 15 +/- 4 to 9 +/- 3 (+/-1 SD) and in Group II from 28 +/- 11 to 16 +/- 7 mm Hg) was observed. The mean arterial pressure decreased in both groups by an average of 9 mm Hg. The changes in the heart rate were minimal. The cardiac output decreased in Group I from 4.4 +/- 1.0 to 3.9 +/- 0.8 1/min (P less than 0.005) whereas in the group with left ventricular failure it increased significantly from a lowered initial value (3.4 +/- 1.0 to 4.0 +/- 1.1 1/min) (P less than 0.001). By using precordial mapping for estimation of ischemic injury in 12 patient sigma ST elevation decreased from 14.6 +/- 11.9 to 12.2 +/- 9.8 mV at a dose of 3 mg/h (P less than 0.03). The dose of 6 mg/h, however, was less effective on the extent of myocardial ischemia than 3 mg/h. Thus, the dosage of nitroglycerin may be critical in respect to infarct size, despite the beneficial hemodynamic effects of the drug.


A comparative study of two methods of measurement of fractional calcium absorption: results in normal individuals and patients with various disturbances of calcium metabolism. Two methods of measurement of fractional calcium absorption have been compared in the same individuals. There is a significant correlation between fractional 47Ca absorption as measured with a blood counting method and with an external counting method after intravenous and oral administration of 47Ca (P less than 0.001). Two varieties of the external counting method, namely with intervals of 2 h and 7 days between the i.v. and oral doses, have been compared. In a group of twenty-one patients with chronic renal failure (not on haemodialysis), 47Ca absorption as measured by external counting was significantly lower than in the control group (P less than 0.001). When put on a dialysis programme a group of six patients showed a significant increase of the fractional 47Ca absorption (P less than 0.01).


Studies on the effects of propanolol on plasma catecholamine levels in patients with essential hypertension. The influence of the beta receptor blocking agent propranolol on plasma catecholamine concentrations was studied in eight patients with essential hypertension. The study was of single blind crossover design. Propranolol given in oral doses ranging from 60 to 240 mg daily for a period of 3 weeks decreased blood pressure and heart rate. The beta-adrenergic blocking agent caused plasma catecholamine levels to increase both at rest and during bicycle exercise. Chromatographical analysis showed that concentrations of noradrenaline as well as of adrenaline rose during treatment with propranolol. However, dopamine-beta-hydroxylase activity in plasma was not altered. Furthermore, the urinary excretion of noradrenaline, adrenaline and 4-hydroxy-3-methoxy mandelic acid did not change during beta receptor blockade. The results are compatible with the assumption that antihypertensive doses of propranolol by decreasing cardiac output cause an activation of the sympatho-adrenal system.


Interlaboratory comparison of radioimmunological parathyroid hormone determination. An inter-laboratory study of serum immunoreactive PTH (iPTH) determinations using standardized sera has been performed in order to check the value of the assays for the diagnosis of hyperparathyroid states. The results demonstrate: (1) that most of the cooperating laboratories (eleven of twelve) were able to discriminate between normal and grossly elevated PTH-values; (2) that direct comparison of values from different laboratories indicates very poor agreement; (3) that PTH values cannot be interpreted without a description of the characteristics of the assay used; (4) that the introduction of standard sera is advantageous and should be undertaken; (5) that determination of iPTH in serum samples is far from being a routine method when compared with radioimmunoassays for hormones like insulin, growth hormone, etc.; the difficulties being due to lack of standardized reagents and peculiarities in the metabolism of the hormone.


Renal glomerular and tubular impairment during strenuous exercise in young women. Creatinine, total protein, albumin and beta2-microglobulin were measured in the urine of fifteen healthy women before and after strenuous short-term exercise. The heavy intermittent load produced an increased urinary excretion of total protein, albumin and beta2-microglobulin, while creatinine was unaffected. The renal clearance of albumin and beta2-microglobulin showed very high values after stopping the exercise. However, 45 min after the end of exercise, total protein returned to initial values while albumin and beta2-microglobulin remained high. The urinary ratio between beta2-microglobulin and albumin is higher in urine collected after exercise than in normal proteinuria. This implies that post-exercise proteinuria is of glomerular and tubular origin.


The Tromsø Heart Study: serum apolipoprotein AI concentration in relation to future coronary heart disease. The concentrations of apolipoprotein AI, a major peptide of high density lipoprotein (HDL), have been measured by immunoelectrophoresis in samples of serum from twelve subjects who subsequently developed a coronary event during 2 years of follow-up and compared with those in serum from sixteen matched control subjects. The mean apolipoprotein AI concentration in the cases was significantly lower than that in the controls, independently of the serum total cholesterol and triglyceride concentrations. There was no significant difference between the cases and controls in mean HDL cholesterol: apolipoprotein AI ratio. Within several case-control pairs, however, the difference in apolipoprotein AI concentration was proportionately much less than that in HDL cholesterol. On discriminant function analysis, apolipoprotein AI concentration was a less powerful predictor of coronary heart disease than was HDL cholesterol.


Plasma phenylethanolamine in hepatic encephalopathy. It has been suggested that amines other than octopamine may be involved in the pathogenesis of hepatic encephalopathy. Plasma phenylethanolamine has been determined by a radioenzymatic method in twenty-six biopsy-proven cirrhotics with or without encephalopathy and in seven normal adults. Phenylethanolamine plasma levels correlated statistically with the presence of liver cirrhosis and severe coma. These results are consistent with the false neurotransmitter hypothesis of hepatic encephalopathy.


Turnover of radiolabelled plasminogen and prothrombin in cirrhosis of the liver. The turnover of purified radiolabelled plasminogen was studied in four patients with cirrhosis of the liver, and that of radiolabelled prothrombin in six patients with cirrhosis of the liver. The cirrhotic patients showed an increased fractional catabolic rate and a decreased synthetic rate, resulting in subnormal plasma levels of plasminogen and prothrombin. The plasma concentration of the two proteins correlated with the synthetic rate, but not with the fractional catabolic rate. Heparin infusion prolonged the shortened half-life of plasminogen in two cirrhotics from 1.25 to 2.10 days and from 1.45 to 1.90 days, and the half-life of prothrombin in three cirrhotics from 2.25 to 2.70 days, from 2.35 to 2.80 days and from 2.40 to 3.70 days. These results indicate that the abnormal turnover of labelled plasminogen and prothrombin in cirrhosis of the liver is due to two mechanisms; increased breakdown, reversible by heparin administration, and impaired synthesis. The decreased plasma levels are, however, mainly caused by decreased synthesis.


Plasma zinc after myocardial infarction. Plasma levels of zinc and albumin have been measured for eight consecutive days in patients suffering chest pain due to myocardial infarction or myocardial ischaemia. A substantial diurnal variation has been found in both controls and patients. Plasma zinc falls after myocardial infarction reaching the lowest level on day 2, thereafter returning to normal by day 8. The lowest plasma zinc levels occur within 48 h of a myocardial infarction and in some patients herald the development of a serious cardiac arrhythmia. Plasma albumin falls progressively after myocardial infarction, being lowest on day 8. The fall in plasma zinc is not totally explained by the fall in plasma albumin.


Allelic exclusion in the B lineage cells of the rabbit. Allelic exclusion of the K polypeptide chain allotypes b4 and b5 of the rabbit exists for the immunoglobulins of almost all plasma cells and of most mature small B lymphocytes. However, both these alleles are expressed simultaneously in rare plasma cells and in an usually large fraction of the lymphoid cells of the bone marrow. Our experiments support the idea that such bone marrow B lineage cells do not always show allelic exclusion.


Antigen presentation in the murine T lymphocyte proliferative response. II. Ir-GAT-controlled T lymphocyte responses require antigen-presenting cells from a high responder donor. The activation of T lymphocytes from poly (Glu60Ala30Tyr10)n (GAT)-primed donors by GAT-pulsed nonimmune spleen cells was shown to require identity at the I-1 subregion of the major histocompatibility complex. However, GAT-primed T lymphocytes from (responder x nonresponder) F1 hybrids could only be stimulated to proliferate by GAT bound to high responder or F1 spleen cells but not by GAT bound to spleen cells from the low responder parent. The failure of spleen cells from low responder parental strains to present GAT was shown not to be due to the presence of suppressor cells in either the antigen-presenting or the responding cell populations. These results indicate that control of antigen-presenting cell-T lymphocyte interactions is one site of Ir gene expression.


T-independent responses in B cell-defective CBA/N mice to Brucella abortus and to trinitrophenyl (TNP) conjugates of Brucella abortus. CBA/N mice have an X-linked immune defect in B lymphocyte function which leads to their inability to respond to several thymus-independent antigens. We report here that these mice and immunologically defective F1 male (CBA/N X DBA/2N) mice can respond to Brucella abortus and to 2,4,6-trinitrophenyl derivatives of Brucella abortus (TNP-BA). These responses can be obtained in vivo and in vitro and are thymus-independent by the criteria that (a) they can be transferred to irradiated recipients by bone marrow cells and anti-Thy-1.2 and complement-treated spleen cells; (b) that nu/nu BALB/c spleen cells respond to TNP-BA in vitro; and (c) that anti-Thy-1.2 and complement-treated (CBA/N X DBA/2N)F1 male spleen cells respond to TNP-BA in vitro. B. abortus and TNP-BA are poor polyclonal B cell activators (PBA) and poor B cell mitogens, unlike lipopolysaccharide which is both a powerful PBA and B cell mitogen. These results therefore indicate that mice with the CBA/N B cell defect can respond to some thymus-independent antigens, namely TNP-BA, and as shown previously, TNP-LPS, although not to other thymus-independent antigens. This, in turn, suggests that thymus-independent antigens may be subdivided on the basis of their ability or inability to stimulate responses by CBA/N B lymphocytes.


The inhibition of cyclo-oxygenase of rabbit platelets by aspirin is prevented by salicylic acid and by phenanthrolines. Salicylic acid, 1,10- and 1,7-phenanthroline prevented inhibition by aspirin of platelet aggregation and of generation of thromboxane A2 due to arachidonic acid, to the ionophore A21387, to thrombin and to collagen. Dithiothreitol, another drug which prevents aggregation and formation of thromboxane A2, but only reversibly, failed to interfere with the inhibition by aspirin. Irreversible inhibition by indomethacin and by the substrate analogue 5,8,11,14-tetraynoic acid was also unaffected by salicylic acid or by 1,10-phenanthroline, which thus probably exert a specific interaction with the aspirin-binding site. Inactivation of platelet cyclo-oxygenase with arachidonic acid led to inhibition of the formation of thromboxane A2 and of aggregation due to arachidonic acid itself and to collagen, but barely affected aggregation by thrombin, even though generation of thromboxane A2 was blocked. Use of salicylic acid and of reversible inhibitors of cyclo-oxygenase may help to unravel the mechanism of inhibition due to other agents.


Visual cortex ablation and thresholds for successively presented stimuli in rhesus monkeys: I. Orientation. Rhesus monkeys were trained to discriminate gratings of different orientation presented one at a time. The smallest difference they could reliably detect was determined before and after removal of either inferotemporal, foveal prestriate or lateral striate cortex. Four animals with inferotemporal lesions had significantly higher thresholds after operation, which did not improve after extensive practice. Two monkeys with lateral striate ablation, and 1 out of 3 animals with foveal prestriate lesions were also impaired, but improved with practice. Monkeys without inferotemporal cortex may have imprecise internal templates for stimulus orientation, in accordance with the categorisation hypothesis of the inferotemporal defect.


Histochemical study of cobalt-induced focal epilepsy. A previous study showed a strong relationship between human focal epilepsy and the presence in the cortex of "activated" astrocytes characterized by an intense activity of dehydrogenases (DH) involved in glucose metabolism and of glutamate DH. Using the semi-chronic model of cobalt-induced experimental focal epilepsy in the rat, we investigated a possible correlation between astrocyte modifications and the chronological development of the epileptic manifestations on the ECoG. After a few days the cobalt-implanted rats present spikes, then sharp waves followed by an electrical crisis and ultimately motor seizures. Activated astrocytes were found in each phase of this evolution. Their number increases with the intensity of the manifestations. There is a close relationship between activated astrocytes and focal epileptic phenomena. At this stage of our study it is clear that the presence of activated astrocytes is not a consequence of seizures. However, it is impossible to say whether the activation is secondary to the hyperactivity of the neurons or directly responsible for the constitution of the epileptic focus. In any case, activated astrocytes provide a new means of localizing an epileptogenic focus.


A horseradish peroxidase-autoradiographic study of parietopulvinar connections in Saimiri sciureus. Descending connections from parietal cortex to pulvinar in squirrel monkey were investigated with the autoradiographic method. Somatosensory areas I (SI) and II (SII) were found to project to the oral (PuO) and medial (PuM) subdivisions of the pulvinar. Projections from the posterior parietal region were recorded in circumscribed areas of PuM and the lateral (PuL) and inferior (PuI) subdivisions of pulvinar. Retrograde labeling with horseradish peroxidase (HRP) demonstrated that rostral parietal cortex including the lateral cortex of SI and the rostral part of area 5 received reciprocal projections from PuO and rostral PuM. Injections of HRP into medial and lateral regions of SI also resulted in labeled cells in PuO and PuM. Within the limitations of the HRP technique, the latter results suggest a direct pathway from pulvinar to primary sensory cortex. The experimental results confirm the accepted view of projections from parieto-temporo-occipital "association" cortex to PuM, PuL and PuI. In addition, reciprocal connections of rostral parietal cortex with PuO and PuM were demonstrated.


[The effect of Ritalin, CB-154 and various drugs of serum GH in acromegalics (author's transl)]. The present study was undertaken to investigate the effects of dopaminergic agents and hypothalamic-releasing hormones on the GH release in acromegalics and normal volunteers. In the methylphenidate (Ritalin) test, nine normal volunteers and four acromegalics were examined. In other tests, five acromegalics were examined. The dopaminergic agents were administered to the subjects orally early in the morning after overnight fasting, but the hypothalamic-releasing hormones were given intravenously at the same time. The results were as follows: (1) The oral administration of 20 mg of Ritalin failed to have any effect on the GH release in normal subjects, and caused a decrease of serum GH level in only one of four acromegalics, the same case which showed an increased GH response to 1-DOPA. In the other three patients, Ritalin did not effect the GH release. Long-term administration of this drug to the acromegalics who showed depression of GH levels resulted in only slightly depressed GH levels. (2) Each oral administration of 2.5 mg of CB-154 or 500 mg of 1-DOPA caused a significant decrease of serum GH level in acromegalics, but CB-154 showed a longer and more stable suppression of serum GH level in all acromegalics. Furthermore, long-term administration of this drug resulted in a continuing inhibited serum GH level without the existence of any serious side effects. It thus seems likely that CB-154 might be a safe and effective drug for the medical treatment of acromegalics. However, while 1-DOPA also caused a significant decrease of serum GH level in four out of five acromegalics, its effect was of much shorter duration. (3) The intravenous administration of 500 microgram of TRH showed a significant increase of serum GH level in four out of five acromegalics, but LH-RH did not have any effect on GH release in any of the acromegalic cases. It seems likely from these results that dopaminergic agents, especially CB-154, may be beneficial for the medical treatment of acromegaly.


Time and dose studies of the effect of cobra venom factor on the in vivo immune response in Galleria mellonella to Pseudomonas aeruginosa. The induced resistance of wax moth larvae vaccinated with formolized Pseudomonas aeruginosa 18 hr prior to challenge with the live organism was significantly inhibited by 0.2 to 1.0 units of cobra venom factor (CVF) per insect given 6 hr after vaccination. Approximately 90% inhibition of the protective response was caused by 1.0 units CVF/insect. Administration of this dose 6 hr before vaccination, with the vaccine, and 6 hr before challenge had no significant effect. The CVF effect was inhibited by heating the CVF preparation at 70 degrees for 30 min.


Early ontogeny of germinal center formation in the chicken. Germinal center formation was studied in the spleen of young chickens immunized in ovo and at the time of hatching. When immunization was performed on day 18 in ovo and on the day of hatching, the first germinal centers were observed at 4 days. This is markedly earlier than in unimmunized chickens, where the first germinal centers appear at the age of 10 days or later. Germinal center formation preceded significant antibody production. The possible role of germinal centers in the generation of immunological memory is discussed in the light of these and earlier observations.


DMBA induced papillomas in congenitally athymic (nude) and hereditarily asplenic (Dh/+) mice: contrasts and comparisons with immunologically intact littermates. Congenitally athymic (nude) and hereditarily asplenic (Dh/+) mice were painted with dimethylbenzanthracene (DMBA) to compare skin tumor development in these immunodeficient animals with their immunologically normal littermate controls. Papillomas were induced in all groups of mice. However, nude and Dh/+ mice were significantly more resistant than their normal littermates to tumor induction. Furthermore, the number of papillomas/mouse and the total tumor incidence were significantly greater in control mice and the latency period for tumor appearance was shorter and the tumor growth rate greater in normal mice compared to their immunodeficient littermates. Finally, nu/+ skin transplanted to nude mice and then painted with DMBA behaved in similar fashion as nude skin. These findings, when discussed in terms of target organs for DMBA, suggest a major role for the immune system in stimulating papilloma induction.


Lymphocyte isolation, rosette formation, and mitogen stimulation in Rhesus monkeys. Over 70% of rhesus monkey peripheral mononuclear cells were isolated on sodium metrizoate-ficoll gradients with greater than 98% purity. Rhesus blood contained 47.8% active E, 58.2% total E, and 30.2% EAC rosette forming cells. Optimal conditions for mitogen studies were determined using phytohemagglutinin, concanavalin A, pokeweed mitogen, lipopolysaccharide and streptolysin O.


5-HT effect and the control of the relaxation of a molluscan smooth muscle (ABRM) of Mytilus edulis L. Morphological differences exist between the arrangement of myofilaments in relaxed EDTA-treated fibres (induced by a 10(-9)M Ca2+ solution, ionic strength 0.28 at pH7.0) and the fibres relaxed by 5-HT added into the same relaxing solution. Complemented by a tension recording of an ABRM preparation, we find the following characteristics during 5-HT relaxation: (i) The distance between centers of thick neighboring filaments is increased. (ii) The thin filaments appear loosely dispersed between the thick ones. These observations suggest that decrease in myofilament interactions is responsible for the plasticizing effects of 5-HT. The sarcolemma appears with some granularity after 5-HT treatment; nexuses and dyads, however, are well defined.


Physiological correlates of maternal separation in surrogate-reared infants: a study in altered attachment bonds. Three pigtail infant monkeys (Macaca nemestrina) were separated from their mothers shortly after birth and raised in social isolation on cloth surrogates. At about 24 weeks of age they were surgically implanted with multichannel biotelemetry systems. Following the collection of baseline behavioral and physiological data, the cloth surrogates were removed for 4 days, then returned. The behavioral and physiological (heart rate, body temperature, sleep patterns) reaction to separation from the surrogate was much less intense than is the case in group-living pigtail infants that are separated from their mothers, suggesting that the attachment bond to a cloth surrogate may be different than the bond to a living mother in a social group. Heart rate mean values and variability were similar in surrogate-reared and mother-reared infants, suggesting a degree of environmental independence in heart rate regulation.


Changes in cortical dendritic branching subsequent to partial social isolation in stumptailed monkeys. Stumptailed monkeys were reared from 1 week after birth to 6 months of age in either a colony condition with the mother or in partial social isolation that allowed visual contact with the colony animals, but not physical contact. At 6 months of age the animals were killed and selected areas of the neocortex stained by the Golgi-Cox method. Relatively nonspiney cells of Layer IV were drawn and analyzed for complexity of dendritic branching. Isolation-reared animals had significantly decreased branching complexity in Motor I cortex when compared to the control animals. A transform of the data that related the number of branches to the number of previous branches showed a slight rearing effect in Somatosensory I cortex with the deprived animals having a lower rate of branching than the controls. We conclude that social isolation also includes a motoric deprivation that could account for these data.


Depression of protective mechanisms against microörganisms in tumor-bearing mice and its restoration by adjuvants. Bacterial growth within 72 hr after an intravenous inoculation with Listeria monocytogenes was enhanced strikingly in the liver of mice, when viable cells of sarcoma-180 were injected subcutaneously into ddN, C3H/He, and BALB/c mice 5 hr before the inoculation. Such an enhanced bacterial growth appears to be attributable to a depressed ability of macrophages to digest engulfed bacteria. Pretreatments with zymosan, killed BCG, or viable BCG prevented such depression in tumor-bearing mice and increased the bactericidal activity in the liver of normal and tumor-bearing mice above the level of non-treated normal mice. Such adjuvants may be useful not only for augmentation of antitumor activity but also for augmentation of antimicrobial activity in tumor-bearing hosts.


Analysis of gynandromorph survivals in Drosophila melanogaster infected with the male-killing SR organisms. The effects of SRO (Sex-ratio organism; previously called SR spirochetes) infection on the viability of gynandromorphs of D. melanogaster were examined. A fraction of gynadromorphs, most of them with small areas of XO tissue, survived, but the majority were not viable when XO nuclei or cells are affected by the infecting SRO. Apparently the replacement of lethal XO nuclei or cells by adjacent XX ones does not take place. Some possibilities are discussed as to the time and the primary site of the lethal action of SRO.


[Mechanisms of genetic regulation of the lactate dehydrogenase isoenzyme spectrum in the erythrocytes of silver-black foxes]. The spectrum of lactate dehydrogenase (LDH) isozymes in erythrocytes of silver foxes was investigated by means of electrophoretic and immunochemical methods. By means of electrophoresis it is shown that differences in LDH isozyme spectrum between the animals homozygous for the gene Ldr-1a and those homozygous for the gene Ldr-1b are most conspisions at the age of 90-100 days of postnatal development. By means of the immunochemical method three groups of animals are distinguished differing in the LDH content in erythrocytes: the animals with a high LDH content (81.0 mcg/ml) and with a low LDH content (54.14 mcg/ml), which are homozygous for the gene Ldr-1b and Ldr-1a respectively, and the animals with intermediate LDH content (64.58 mcg/ml), which are heterozygous. The data obtained suggest that the effect of the gene Ldr-1a is associated with the decrease of the quantity of A subunits of LDH. It is assumed that the mechanism of the gene Ldr-1a action is realized either by means of the decrease of the synthesis of the A LDH subunits, or by means of the increase of the rate of their degradation.


[Genetic blood markers in arthritic diseases]. In the course of studying patients affected with arthritic diseases (bronchial asthma, sugar diabetes) a relationship between those diseases and the blood groups of the Hp and MN systems was established. Patients with bronchial asthma more frequently than in the controls belonged to the Hp 2-2 type and to blood groups MN and O (I), whereas the patients affected with sugar diabetes are usually of the Hp 2-1 type and belong to the belong to the blood groups MN and A(II). The investigation of patients affected with other diseases having a pathogenesis similar to that of bronchial asthma or of diabetes and the observation of healthy persons after prophylactic inoculations as well as the study of sugar metabolism in healthy persons all confirm the relationship of Hp, MN and ABO antigens with arthritic diseases.


[Sarcosporidiosis of roe deer (Capreolus capreolus L.)]. Examination of 197 specimens of roe deer, of 8 regions of Bohemia, both males and females of various age, revealed sarcosporidiosis in 76% of animals. The incidence of sarcosporidiosis was higher in the regions where the landscape was not much influenced by cultivating. Two types of microscopical sarcocysts, thin-walled and thick-walled ones, were found for the first time in roe deer. The thick-walled cysts were observed in one region only. Then, the sarcosporidiosis in roe deer seems to be caused by two species of coccidia. In almost 14% of positive cases of sarcosporidiosis, a pathological reaction of the infected musculature was observed.


Does money spent on health care really improve U.S. health status? Whether increased expenditures for health care substantially improve the overall health care of the U.S. population poses a critical question for the formulators of a national health policy. Concentrating exclusively on the impact of the Medicare program on the health status of the aged poor, the author examines HEW data and assesses the linkage between increased expenditures and improved health.


Rh disease: how near the end?

Rh disease: how near the end? Hemolytic disease of newborns is now a preventable disease, 10 years after the introduction of routine postpartum administration of hyperimmune globulin to Rh-negative mothers. Careful adherence to clinical and administrative guidelines, such as those described in this article, can help ensure that every susceptible woman is treated, so that the disease can be eliminated in another decade.


GM1-gangliosidosis: accumulation of ganglioside GM1 in cultured skin fibroblasts and correlation with clinical types. Uptake of radioactivity from 14C-galactose into gangliosides by cultured skin fibroblasts was studied. GM3 was the major ganglioside in control human fibroblasts. An increase of GM1 was demonstrated in GM1-gangliosidosis fibroblasts. The degree of GM1 accumulation was correlated with the clinical types of this disease. The fibroblasts from an infantile-type patient showed a marked increase of GM1. In late-onset types the amount of total gangliosides was only slightly increased, but the distribution of individual gangliosides was definitely abnormal; a relative increase of GM1 was demonstrated in these cases. GM1 beta-galactosidase activities were not detectable in either infantile or late-onset cases.


Depression of contact sensitivity by enhancement of suppressor cell activity in Pseudomonas aeruginosa-injected mice. Heat-killed Pseudomonas aeruginosa depresses contact sensitivity to oxazolone in C56BL/6 mice. The draining lymph nodes and spleens of mice exhibiting an impaired reactivity to oxazolone contain a cell population capable of depressing the response to oxazolone of recipients sensitized immediately before cell transfer. The suppressive activity of these cells appears to be antigen specific, since they do not affect the response to picryl chloride and because they do not arise in P. aeruginosa-injected but not oxazolone-sensitized mice. These suppressor cells occur in the draining lymph nodes and spleen at 3 and 4 days after sensitization, respectively, and have precursors sensitive to cyclophosphamide. It is concluded that P. aeruginosa depresses contact sensitivity to oxazolone by enhancing the suppressor cell activity of the regulatory cells which arise during conventional sensitization.


Role for activated macrophages in resistance against Trichinella spiralis. To determine whether activated macrophages are important in resistance against the intestinal phase of nematode parasites, we studied Trichinella spiralis infections in mice with normal macrophages and in mice with macrophages activated by either chronic Toxoplasma gondii or acute Listeria monocytogenes infections. The peak T. spiralis adult worm burden in the intestines of normal C57BL/6 or Swiss Webster mice occurred from 6 to 14 days after infection. Subsequent expulsion of worms from the intestines occurred from 8 to 20 days after infection. C57BL/6 mice chronically infected with T. gondii and then challenged with T. spiralis larvae had significantly lower peak intestinal worm burdens (P < 0.05) than normal C57BL/6 mice similarly challenged. Swiss Webster mice infected 7 or 13 days earlier with L. monocytogenes and then challenged with T. spiralis larvae had significantly lower peak worm burdens (P < 0.01) than uninfected mice. The time of expulsion of adult worms was not affected by either infection. Swiss Webster mice infected 42 days earlier with L. monocytogenes (i.e., possessing lymphocytes sensitized to L. monocytogenes but not possessing activated macrophages) did not have a lower worm burden than uninfected mice. Serum factors (e.g., antibody) did not appear to play a role because normal mice injected with serum from L. monocytogenes-infected mice had worm burdens similar to those of mice injected with normal serum. The histopathology of intestines of mice infected with T. gondii or L. monocytogenes was the same as that of normal mice. When T. spiralis larvae were incubated with normal macrophages or macrophages from T. spiralis-infected mice in vitro for 24 h, the number of larvae with adherent T. spiralis macrophages was significantly (P < 0.005) greater than the number of larvae with adherent normal macrophages. These studies suggest a role for activated macrophages in resistance to T. spiralis.


Mechanisms of immunity in typhus infection: some characteristics of Rickettsia mooseri infection of guinea pigs. Rickettsia mooseri infection has been studied in syngeneic guinea pigs inoculated intradermally with the objective of developing a model for the study of immune mechanisms. Characterization of infection included the following: a study of replication, dissemination, and clearance of rickettsiae; measurement of the antibody response with different rickettsial antigens and tests; and attempts to measure the cell-mediated immune response using the correlate of delayed-type hypersensitivity skin reactions. Following intradermal inoculation, rickettsiae replicate locally and then spread to the draining lymph nodes and subsequently cause systemic infection. Spread to draining lymph nodes occurred before the appearance of circulating antibody, whereas systemic infection occurred afterwards. Two distinct patterns of acquired resistance developed. The first was marked by a cessation of rickettsial growth within a given organ and the second by a clearance of rickettsiae. The duration of each of these phases differed markedly from one organ to another. Delayed-type hypersensitivity was not demonstrated by skin testing.


Some ultrastructural effects of persistent infections by the rickettsia Coxiella burnetii in mouse L cells and green monkey kidney (Vero) cells. Mouse fibroblasts (L-929) and Vero (green monkey kidney) cells were infected with the rickettsia Coxiella burnetti, and persistent infections developed and were studied over a 6- to 10-month period. Ultrastructural comparisons were made between the two infected cell types, and both were tested cytochemically for the presence of acid phosphatase, a marker enzyme of lysozymes. Rickettsiae were always observed within vacuoles, and some infected L cells showed flattened endoplasmic reticulum as compared with uninfected cells. Rickettsiae in Vero cells were most often seen in vacuoles containing whorls of membranes ("myelin configurations") which were also seen in uninfected cells. Rickettsiae in Vero cells were pleomorphic, with acid phosphatase reaction product in their periplasmic space. This suggests either rickettsial degradation by lysosomal enzymes which penetrated the cell envelope or a penetration after the rickettsiae were dead. Vacuoles of infected Vero cells showed much more reaction product than that in infected L cells, and most rickettsiae in L cells had a normal appearance and showed no reaction product in their periplasmic space.


Serological cross-reactivity among Sporothrix schenckii, Ceratocystis, Europhium, and Graphium species. Ethanol-precipitable culture filtrate antigens of 100 strains of 75 species of the Sporothrix-Ceratocystis-Europhium-Graphium complex and 1 species of Botrytis were examined for neutral sugar components and for serological cross-reactivity with S. schenckii rabbit antiserum and human sporotrichosis sera by capillary precipitin and double immunodiffusion assay. Results revealed that cross-reactive species (60 of 77, ca. 80%) produced exoconidial forms and rhamnose- and mannose-containing polysaccharides and included Ceratocystis, the three known Europhium, and several Graphium-form species. Endoconidial-form Ceratocystis species did not cross-react.


Incomplete inhibition of the immunoglobulin G-binding behavior of nitrated protein A. Nitration of protein A with tetranitromethane incompletely blocks its Fc-binding ability for immunoglobulin G when measured by more sensitive methods of immunoprecipitation and when tested in the enzyme-linked immunosorbent assay. In addition, binding of protein A and blockage by nitration is heterogeneous and depends on the species origin of the immunoglobulin G.


Some aspects of pharmacokinetic and biotransformation differences in humans and mammal animals. The erroneous tendency to set pharmacokinetics and biotransformation of drugs in man against results obtained in animals as a different homogenous group is discussed. Species differences have mainly a quantitative character, such as the body weight to surface ratio, organ weight to body weight ratio, presence of differing fat deposits, etc. Qualitative differences are not so common. The great importance of a better knowledge of these problems for the prediction of drug effects in man from results obtained in animals is emphasized.


A zinc-deficiency dermatitis in patients on total parenteral nutrition. Three patients on home total parenteral nutrition utilizing a synthetic amino acid solutions developed a dermatitis consistent with acrodermatitis enteropathica after 30, 43 and 62 days of therapy. The dermatitis resolved after changing to casein hydrolysate in 2 patients and after full oral alimentation in the third. Although measured serum zinc levels were repeatedly normal to elevated in all patients, sufficient retrospective data analysis combined with literature review on this subject, clearly implicates zinc deficiency as the etiology for their dermatitis. Adequate zinc supplementation of synthetic amino acid solutions is essential in order to avoid this complication.


Studies on the duration of local anesthesia. Effects of volume and concentration of a local anesthetic solution on the duration of dental infiltration anesthesia. In animals and humans the rate of resorption and the duration of the anesthesia have been investigated, using mepivacaine in different concentrations and volumes without addition of a vasoconstrictor. A linear relationship between duration of anesthesia and the logarithm to the concentration of mepivacaine was found.


Myoxomas of the jaws.

Myoxomas of the jaws. A retrospective study on 15 patients with myxomas of the jaws was carried out. Follow-up information was obtained in 10 patients. Clinical, radiologic, and microscopic features were examined and the results of extensive resection versus conservative surgery were evaluated. The results seem to be in favor of the latter. Four cases are presented in detail.


Conservative surgical treatment of mandibular ameloblastomas. The results of a follow-up investigation of 12 patients with ameloblastomas of the mandible, treated by conservative surgical means, are presented. The age of the patients at the time of operation ranged from 8 to 77 years, the average age being 41 years. Seven of the tumors had a unilocular and five a multilocular appearance. The operation technique varied somewhat with the individual case. An attempt was made in all cases to remove the tumor in toto. In cases with multilocular appearance or where the tumor was in connection with the surface epithelium, a resection of the roof of the lesion was made. Microscopic examination of the tumors revealed that they all could be classified as plexiform or folicular but with various degrees of differentiation. Seven of the patients were observed for more than 5 years, the others for less. Three instances of recurrence were diagnosed and were in all cases less than 1 cm in diameter. The recurrences were found 1, 3, and 20 years after the primary treatment. Although the number of patients and the observation period are limited, the preliminary results seem encouraging.


Lipomas of the oral cavity.

Lipomas of the oral cavity. The clinical and histopathologic characteristics of oral lipomas are reviewed. A series of six patients undergoing surgical treatment for oral lipomas at the University of Bergen, Department of Oral Surgery during the period 1969--1976 is presented. The characteristics of the six tumors are described and the findings compared with those of previously reported cases.


Resorption of the crown in impacted maxillary canine. A clinical, radiographic and histologic study. Thirty-six maxillary impacted canines which showed coronary resorption were selected out of 252 impacted canines. Clinical, radiographic and histologic observations were performed. In the majority of the cases (31) a local inflammatory factor was present and only in five cases was the resorption idiopathic. The incidence of resorption was highest in the later decades of life. In all cases bone replacement was found.


Sialochemistry of KCS patients.

Sialochemistry of KCS patients. Sialochemistry was used to detect salivary gland involvement in KCS patients. Significantly elevated IgA was found in 52% of the patients, potassium in 48% and sodium in 32%. No correlation between the grade of Schirmer's test and the extent of changes in salivary composition was observed. The results indicate that salivary IgA, K and Na concentrations can be used for detection of salivary gland involvement of KCS patients.


Ameloblastoma with gingival ulcer.

Ameloblastoma with gingival ulcer. Ameloblastoma usually presents as a painless swelling of the jaw, but the chief complaint is rarely of ulceration. A case of ameloblastoma with a complaint of gingival ulcer in a 75-year-old Japanese male who was treated by an en bloc removal of the tumor with gingival ulcer is described and discussed. There is no evidence of recurrence, and the postoperative course was uneventful.


Metastatic choriocarcinoma involving the gingiva. A case of metastatic choriocarcinoma involving the mandible is reported. This case is analyzed for the potentiality of immunologic defense by utilizing tuberculin test and scoring the rate of blast formation of peripheral blood lymphocytes by phytohemagglutinin, resulting in a greatly decreased ability of cellular immunity.


Occlusal concepts for the orthognathic surgeon. A review of modern occlusal concepts is of great value to the orthognathic surgeon in treating dentofacial deformities. A thorough knowledge of occlusion is essential in the correct diagnosis and treatment of patients requiring surgical alterations in their occlusal relationships. Following these surgical alterations, a critical postoperative evaluation is important in determining the timing and need for occlusal equilibration and/or restorative measures.


Dental management of the irradiated patient. There is an increasing number of patients receiving radiation therapy for oral malignancies. In many malignant tumors, radiation is often the treatment of choice, while in others it may be used in conjunction with surgery or chemotherapy. There are inherent dental and oral problems associated with radiation therapy. It is the purpose of this paper to deal briefly with the physical principles and the biological basis of radiation therapy. In addition, the specific radiation effects on oral mucous membranes, taste buds, salivary glands, bone, the periodontium and teeth will be discussed. Radiation complications in edentulous patients, and in particular the problems of wearing dentures in such patients will be evaluated. An approach to the problem of dental extractions and other dental treatments in the pre- and post-irradiated patient is suggested.


[Role of the digestive flora in adaptation to lactose consumption in rats]. Part of the digestive flora in the adaptation to lactose consumption. Lactase activity was determined with adult Wistar rats. Some of which were accustomed to lactose since weaning. For this purpose, the tissue of jejunum, the flora of the ileum, the caecum and the large intestine were examined. When the rats received a high lactose diet, the lactase of the jejunum is more active because of both the intestinal tissue development and a higher production of enzyme by protein unit. But the main source of lactase is the digestive flora of the animal: a lactase flora develops in the ileum, the large intestine and mainly the caecum. The caecum lactase represents about half of the total lactase activity. The flora of the animal which has not consumed lactose since weaning can develop a noticeable lactase activity after 7 hr of incubation in presence of lactose. It is also the caecum flora which shows the greatest capacity of adaptation to lactose.


Leu-enkephalin-like material in nerves and enterochromaffin cells in the gut. An immunohistochemical study. The distribution and cellular localization of leu-enkephalin in the gut and pancreas was studied by immunohistochemistry using two different antisera, one specifically directed against leu-enkephalin and the other cross reacting with met-enkephalin. The results were identical with both antisera. In all species examined, enkephalin-immunoreactive material was found in nerves of the smooth muscle, particularly numerous in the myenteric plexus. Here, immunoreactive nerve cell bodies were observed occasionally. In addition, enkephalin-immunoreactive material was demonstrated in gut endocrine cells of chicken, mouse, rat, pig and monkey but not of guinea pig, cat and man. Enkephalin cells were detected also in the exocrine parenchyma of the porcine pancreas. They were rare in the gut of mouse, rat and monkey but numerous in the antrum and duodenum of pig where they were identified as 5-hydroxytryptamine-storing enterochromaffin cells. The enkephalin-containing cells of the porcine antrum and duodenum were defined ultrastructurally by the consecutive semithin/ultrathin section technique. The ultrastructural features were typical of enterochromaffin cells, the most characteristic ones being the irregular shape and high electron density of the cytoplasmic granules. The immunoreactive material was confined to the cytoplasmic granules.


Effect of rapid decompression and associated hypoxic phenomena in euthanasia of animals: a review. Documentation in the literature indicates that death is as painless following the induction of hypoxia by rapid decompression as by other methods that lead to hypoxia, such as exposure to high altitude, carbon monoxide, and inert gases (nitrogen, xenon, and krypton). Many of the signs and symptoms of hypoxia are the same as those for alcoholic intoxication and inert gas narcosis. Moreover, there is good evidence that analogous relationships or mechanisms may exist for hypoxia, inert gas narcosis, and anesthesia.


A maturation change detected in the semilunar cartilages with the scanning electron microscope. The surface of rabbit, cat, monkey and human semilunar cartilages was examined with the scanning electron microscope. A common feature was the occurrence of numerous ridges, undulations and furrows on the surface, but this was thought to be due to marked shrinkage and distortion of cartilage not firmly attached to bone. Humps were seen on the semilunar cartilages of young animals, but pits occurred in adults. This is thought to reflect a maturation change. Humps were seen in a young human semilunar cartilage, but pits were not seen in adult specimens. It is not clear whether pits are truly absent or just masked by the severe ridging produced during the preparation of large human specimens.


Alkalimetric microassay of cephalosporins. Alkalimetric pH-stat titrations of cephalosporin C, cephacetril and their deacetyl derivatives using an acetyl esterase and beta-lactamase are described. The esterase was used to assay highly purified samples of cephalosporin C and cephacetril, and also to prepare analytically defined solutions of the deacetyl cephalosporins. Lactamase-catalyzed hydrolysis of the parent compounds was then found to generate exactly 2 equivalents of acid per mole; that of the deacetyl derivatives exactly 1 equivalent.


Lipid and protein composition of membranes of Bacillus megaterium variants in the temperature range 5 to 70 degrees C. Membranes were prepared from four temperature range variants of Bacillus megaterium: one obligate thermophile, one facultative thermophile, one mesophile, and one facultative psychrophile, covering the temperature interval between 5 and 70 degrees C. The following changes in membrane composition were apparent with increasing growth temperatures: (i) the relative amount of iso fatty acids increased and that of anteiso acids decreased, the ratio of iso acids to anteiso acids being 0.34 at 5 degrees C and 3.95 at 70 degrees C, and the pair iso/anteiso acids thus seemed to parallel the pair saturated/unsaturated acids in their ability to regulate membrane fluidity; (ii) the relative/unsaturated acids in their ability to regulate membrane fluidity; (ii) the relative amount of long-chain acids (C16 to C18) increased fivefold over that of short-chain acids (C14 and C15) between 5 and 70 degrees C; (iii) the relative amount of phosphatidylethanolamine increased, and this phospholipid accordingly dominated in the thermophilic strains, whereas diphosphatidylglycerol was predominant in the two other strains; and (iv) the ratio of micromoles of phospholipid to milligrams of membrane protein increased three-fold between 5 and 70 degrees C. Moreover, a quantitative variation in membrane proteins was evident between the different strains. Briefly, membrane phospholipids with higher melting points and packing densities appeared to be synthesized at elevated growth temperatures.


Bacillus subtilis spore coats: complexity and purification of a unique polypeptide component. Extensively washed, dormant spores of Bacillus subtilis were disrupted with glass beads in buffer at pH 7 in the presence of protease inhibitors. Approximately 31% of the total spore protein was soluble, and another 14% was removed from the insoluble fraction by hydrolysis with lysozyme and washing with 1 M KCl and 0.1% sodium dodecyl sulfate. The residual spore integuments comprised 55% of the total spore proteins and consisted of coats and residual membrane components. Treatment of integuments with sodium dodecyl sulfate and reducing agents at pH 10 solubilized 40% of the total spore protein. Seven low-molecular-weight polypeptide components of this solubilized fraction comprised 27% of the total spore protein. They are not normal membrane components and reassociated to form fibrillar structures resembling spore coat fragments. The residual insoluble material (15% of the total spore protein) was rich in cysteine and was probably also derived from the spore coats. A solubilized coat polypeptide of molecular weight 12,200 has been purified in good yield (4 to 5% of the total spore protein). Five amino acids account for 92% of its total amino acid residues: glycine, 19%; tyrosine, 31%; proline, 23%; arginine, 13%; and phenylalanine, 6%.


Genetic and biochemical characterization of kirromycin resistance mutations in Bacillus subtilis. Spontaneous mutations causing resistance to the EF-Tu-specific antibiotic kirromycin have been isolated and mapped in Bacillus subtilis. Three-factor transductional and transformational crosses have placed the kir locus proximal to ery-1 and distal to strA (rpsL) and several mutations affecting elongation factors EF-G and EF-Tu, in the order: cysA strA [fus-1/ts-6(EF-G)] [ts-5(EF-Tu)] kir ery-1 spcA. Purified EF-Tu from mutant strains is more resistant to kirromycin as measured by in vitro protein synthesis and also shows a more acidic isoelectric point than wild-type EF-Tu. This indicates that the kir locus is the genetic determinant (tuf) for EF-Tu and that there is a single active gene for this enzyme in B. subtilis.


Phycobilisomes in spheroplasts of Anacystis nidulans. Phycobilisomes in Anacystis nidulans can be seen more readily in spheroplasts than in cells with intact walls.


Kasugamycin-resistant mutants of Bacillus subtilis. Kasugamycin-resistant mutants of Bacillus subtilis were isolated and classified into two groups, one of which had resistance to kasugamycin in in vitro protein synthesis and mapped in the ribosomal region. The other group had no resistance to kasugamycin in in vitro protein synthesis and had weak cross-resistance to gentamicin and kanamycin. Neither group could sporulate in the presence of kasugamycin.


Comparative study of DNA methylation in three unicellular eucaryotes. We have analyzed the nature/content of methylated bases in the nuclear DNA of three unicellular eucaryotes. The pattern of methylation was different for each of the three organisms studied: Saccharomyces cerevisiae contained only 5-methylcytosine; Tetrahymena pyriformis contained only N6-methyladenine; and Chlamydomonas reinhardi contained both modified bases.


DNA synthesis in competent Bacillus subtilis cells. Competent cells of Bacillus subtilis incorporate degradation products from transfecting DNA into their chromosomal DNA. The sensitivity of this incorporation to inhibitors of bacterial DNA synthesis [phage infection or 6-(p-hydroxyphenylazo)-uracil] suggests that semiconservative DNA synthesis can occur in competent cells.


Early stages in Bacillus subtilis transformation: association between homologous DNA and surface structures. The addition of ethylenediaminetetraacetate to competent cultures of Bacillus subtilis irreversibly inhibited the transformability as well as the cellular binding of DNA. Our results show that the inhibition of DNA binding by ethylenediaminetetraacetate in whole cells, protoplasts, and membrane vesicles is mainly due to a permanent alteration of the DNA receptors. Transformation absolutely requires free magnesium ions, whereas DNA binding is a magnesium-independent step. In contrast to ethylenediaminetetraacetate, the absence of Mg2+ does not irreversibly affect the capacity of the competent cells to be transformed DNA-binding receptors located at the cell surface remain associated with the plasma membrane after protoplasting and after isolation of membrane vesicles. A Mg2+-dependent endonucleolytic activity associated with the membrane appears to be responsible for the lower levels of binding by protoplasts in the presence of this ion.


Comparison of spontaneous, UV-induced, and nitrosoguanidine-induced mutability to drug resistance in myxobacteria. The UV survival curves of different strains of myxobacteria exhibited shoulders; in the case of Polyangium luteum, an unusual double shoulder appeared. Repair inhibitors like acriflavine, caffeine, and coumarin reduced the survival of UV-irradiated cells if the drugs were incorporated in the post-irradiation plating medium. The shoulders were reduced, but the final inactivation slopes were not affected by the repair inhibitors. Those strains that were resistant to UV were also more resistant to being killed by nitrosoguanidine. A variety of drug-resistant mutants occurred. The spontaneous mutation frequencies to drug resistance varied with the drug and the strain used. Drug-resistant mutants were inducible by UV irradiation and nitrosoguanidine. The UV mutability of Myxococcus xanthus was high compared to Cystobacter sp. However, the nitrosoguanidine mutability of M. xanthus was low compared to the other strains.


In vivo titration of araC protein.

In vivo titration of araC protein. The requirement for araC protein in the induction of the araBAD operon was investigated. Strains of Escherichia coli carrying an araC(Am) mutation and temperature-sensitive amber suppressors were used to vary the intracellular level of araC protein. The levels of araC protein studied ranged from 0.007 to 1.8 times the normal amount. The results indicate that the normal level of araC protein is just sufficient to provide maximal expression of the araBAD operon.


Isolation and characterization of Bacillus megaterium mutants containing decreased levels of spore protease. A proteolytic activity present in spores of Bacillus megaterium has previously been implicated in the initiation of hydrolysis of the A, B, and C proteins which are degraded during spore germination. Four mutants of B. megaterium containing 20 to 30% of the normal level of spore proteolytic activity have been isolated. Partial purification of the protease from wild-type spores by a reviewed procedure resulted in the resolution of spore protease activity on the A, B, and C proteins into two peaks--a major one (protease II) and a minor one (protease I). The protease mutants tested lacked active protease II. All of the mutants exhibited a decreased rate of degradation of the A, B, and C proteins during spore germination at 30 degrees C, but degradation of the proteins did occur. Degradation of the A, B, and C proteins during germination of the mutant spores was decreased neither by blockade of ATP production nor by germination at 44 degrees C. Initiation of spore germination was normal in all four mutants, and all four mutants went through outgrowth, grew, and sporulated normally in rich medium. Similarly, outgrowth of spores of two of the four mutants was normal in minimal medium at 30 degrees C. In the two mutants studied, the kinetics of loss of spore heat resistance and spore UV light resistance during germination were identical to those of wild-type spores. This indicates that the A, B, and C proteins alone are not sufficient to account for the heat or UV light resistance of the dormant spore.


Transport by the lactose permease of Escherichia coli as the basis of lactose killing. Lactose killing is a peculiar phenomenon in which 80 to 98% of the Escherichia coli cells taken from a lactose-limited chemostat die when plated on standard lactose minimal media. This unique form of suicide is caused by the action of the lactose permease. Since uptake of either lactose or galactose by the lactose permease caused death, the action of rapid transport across the membrane must be the cause of the phenomenon. Alternative causes of lactose killing, such as accumulation of toxic metabolic intermediates or action of the beta-galactosidase, have been eliminated. It is proposed that rapid uptake of sugars by the lactose permease disrupts membrane function, perhaps causing collapse of the membrane potential.


Dark hexose metabolism by photoautotrophically and heterotrophically grown cells of the blue-green alga (Cyanobacterium) Nostoc sp. strain Mac. Photoautotrophically grown cells of the blue-green alga (cyanobacterium) Nostoc sp. strain Mac assimilated and oxidized both glucose and fructose in the dark at different rates. The rate of fructose metabolism in these cells could be stimulated by casein hydrolysate, the effect being most pronounced at low sugar concentrations. This stimulation was not seen in cells grown heterotrophically in the dark, suggesting that it is a transitory phenomenon which disappears during the autotrophy-heterotrophy growth transition. The stimulation of fructose assimilation by casein hydrolysate was abolished by chloramphenicol or streptomycin, suggesting there are rate-limiting steps in protein biosynthesis in the dark that ultimately lead to inhibition of fructose uptake. Glucose metabolism did not show these phenomena, indicating there are differences in the metabolism of the two sugars.


Transfer-deficient mutants of the narrow-host-range plasmid R91-5 of Pseudomonas aeruginosa. Three methods have been successful in the isolation of transfer-deficient mutants of the narrow-host-range R plasmid R91-5 of Pseudomonas aeruginosa: (i) selection for donor-specific phage resistance; (ii) direct screening after mutagenic treatment with either ethyl methane sulfonate or N-methyl-N'-nitro-N-nitrosoguanidine; (iii) in vitro mutagenesis of plasmid DNA by hydroxylamine followed by transformation and direct screening. The majority of transfer-deficient mutants were donor-specific phage resistant, supporting the view that sex pili and other surface components are essential for conjugal transfer (since the phages PRD1 and PR4 adsorb to these sites). Some of the transfer-deficient mutants were also unable to inhibit the replication of phage G101 or lost entry exclusion or both phenotypes. The ability to revert these pleiotropic mutants to wild type implicates the latter two functions in R91-5 transfer. Suppressor mutations in P. aeruginosa enabled the detection of suppressor-sensitive, transfer-deficient mutants. Such mutants should prove useful in conjugational complementation tests for the identification of the transfer cistrons of R91-5.


Aspartate transcarbamylase synthesis ceases prior to inactivation of the enzyme in Bacillus subtilis. Aspartate transcarbamylase is synthesized during exponential growth of Bacillus subtilis and is inactivated when the cells enter the stationary phase. This work is a study of the regulation of aspartate transcarbamylase synthesis during growth and the stationary phase. Using specific immunoprecipitation of aspartate transcarbamylase from extracts of cells pulse-labeled with tritiated leucine, we showed that the synthesis of the enzyme decreased very rapidly at the end of exponential growth and was barely detectable during inactivation of the enzyme. Synthesis of most cell proteins continued during this time. When the cells ceased growing because of pyrimidine starvation of a uracil auxotroph, however, synthesis and inactivation occurred simultaneously. Measurement of pools of pyrimidine nucleotides and guanosine tetra- and pentaphosphate demonstrated that failure to synthesize aspartate transcarbamylase in the stationary phase was not explained by simple repression by these compounds. The cessation of aspartate transcarbamylase synthesis may reflect the shutting off of a "vegetative gene" as part of the program of differential gene expression during sporulation. However, aspartate transcarbamylase synthesis decreased normally at the end of exponential growth at the nonpermissive temperature in a mutant strain that is temperature-sensitive in sporulation and RNA polymerase function. Cessation of aspartate transcarbamylase synthesis appeared to be normal in three other temperature-sensitive RNA polymerase mutants and in several classes of spo0 mutants.


Spore coat protein synthesis in cell-free systems from sporulating cells of Bacillus subtilis. Cell-free systems for protein synthesis were prepared from Bacillus subtilis 168 cells at several stages of sporulation. Immunological methods were used to determine whether spore coat protein could be synthesized in the cell-free systems prepared from sporulating cells. Spore coat protein synthesis first occurred in extracts from stage t2 cells. The proportion of spore coat protein to total proteins synthesized in the cell-free systems was 2.4 and 3.9% at stages t2 and t4, respectively. The sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis patterns of immunoprecipitates from the cell-free systems showed the complete synthesis of an apparent spore coat protein precursor (molecular weight, 25,000). A polypeptide of this weight was previously identified in studies in vivo (L.E. Munoz, Y. Sadaie, and R.H. Doi, J. Biol. Chem., in press). The synthesis in vitro of polysome-associated nascent spore coat polypeptides with varying molecular weights up to 23,000 was also detected. These results indicate that the spore coat protein may be synthesized as a precursor protein. The removal of proteases in the crude extracts by treatment with hemoglobin-Sepharose affinity techniques may be preventing the conversion of the large 25,000-dalton precursor to the 12,500-dalton mature spore coat protein.


Supercoiled DNA folded by nonhistone proteins in cultured mouse carcinoma cells. Upon gentle lysis of exponentially growing mouse carcinoma cells FM3A by sodium dodecyl sulfate, DNA was released as a "DNA-protein complex" in a folded conformation. No histones could be detected in the DNA-protein complex. The proteins bound to DNA were found to be composed of several kinds of nonhistone proteins with a molecular weight range of 50,000 to 60,000; they appear to play a key role in stabilizing and maintaining the compact and folded structure of the complex. Removal of the proteins by Pronase or 2-mercaptoethanol produced a more relaxed structure sedimenting about half as fast as the original complex in a neutral sucrose gradient. DNA in the folded complex is supercoiled, as indicated by the characteristic biphasic response of its sedimentation rate to increasing concentration of various intercalating agents, actinomycin D, ethidium bromide and acriflavine, with which the cells were treated before lysis. Pronase- or 2-mercaptoethanol-treated relaxed DNA still possessed the characteristic of closed-circular structure as judged from its response to intercalating agents. Nicking with gamma-ray or 4NQO broke these superhelical turns and relaxed the folded complex to slower sedimenting forms equivalent to the relaxed DNA obtained on treatment with Pronase or 2-mercaptoethanol. Viscometric observations of DNA-protein complex were consistent with the above results. A tentative model for the structure of this DNA-protein complex is proposed in which supercoiled DNA is folded into loops by several kinds of nonhistone proteins. Autoradiographic examination of the complex appeared to support this model.


Separation and characterization of the outer membrane of Pseudomonas aeruginosa. A method is described for the preparation of outer and cytoplasmic membranes of Pseudomonas aeruginosa, and the outer membrane proteins characterized. Isolated outer and cytoplasmic membranes differed markedly in the content of 2-keto-3-deoxyoctonate (lipopolysaccharide) and phospholipid as well as in the localization of certain enzymes (NADH oxidase, succinate dehydrogenase, D-lactate dehydrogenase, malate dehydrogenase, and phospholipase), and also in the microscopic morphology. The outer membrane preparation showed activity neutralizing a certain bacteriocin or bacteriophages, whereas the cytoplasmic membrane preparation showed no neutralizing activity. The protein composition of membrane preparations from five different strains of P. aeruginosa [P14, M92 (PAO1), PAC1, P15, and M2008 (PAT)] were determined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. More than 50 protein bands were detected in the cytoplasmic membrane preparation. The protein compositions of outer membranes from the five different strains were very similar: at least 6 major bands were found (apparent molecular weights: Band D, 50,000; band E, 45,000; band F, 33,000; bands G and H, 21,000; and band I, 8,000). The protein composition of outer membranes was affected by some physiological growth conditions. Some features of major outer membrane proteins were also studied. Band F showed anomalous migration on SDS polyacrylamide gel electrophoresis depending on the solubilizing conditions or pretreatment with TCA. Band I seemed to be a protein analogous to the lipoprotein which had been found in the outer membrane of Escherichia coli.


Biogenesis of endoplasmic reticulum membrane in rat liver cells. III. Biosynthesis of NADPH-cytochrome c reductase and cytochrome b5 by loosely-bound ribosomes. Membrane-bound ribosomes were separated into two distinct classes (loosely-bound and tightly-bound ribosomes) by treatment with 0.6 M KCl, 1 mM puromycin, 0.05% DOC, or 10 mM EDTA. It was also confirmed that any one of these reagents except for EDTA dissociated the same class of ribosomes from the membrane. A population of lighter microsomal vesicles was formed from rough microsomes upon the dissociation of loosely-bound ribosomes by treatment with these chemicals. Rough microsomes were subfractionated into lighter and heavier fractions, L-rMs and H-rMs, by centrifugation using a discontinuous gradient of sucrose consisting of 1.3 M, 1.5 M, and 2.1 M solutions. It was found that L-rMs was rich in loosely-bound ribosomes, whereas H-rMs contained a high proportion of tightly-bound ribosomes. It is likely that loosely-bound and tightly-bound ribosomes are heterogeneously distributed among rough microsomal vesicles. Loosely-bound ribosomes and tightly-bound ribosomes synthesize different kinds of proteins. Two microsomal membrane proteins, NADPH-cytochrome c reductase and cytochrome b5, were exclusively synthesized by loosely-bound ribosomes, whereas serum albumin, which is a major component of the secretory proteins of hepatocytes, was synthesized only by tightly-bound ribosomes. Since the nascent peptides of NADPH-cytochrome c reductase and cytochrome b5 are released from bound ribosomes to the cytoplasmic surface of endoplasmic reticulum, while those of secretory proteins are discharged into the lumen across the membrane, the strength of the association between ribosomes and microsomal membrane seems to be correlated with the direction of release of nascent peptides.


Spore coat protein of Bacillus subtilis. Structure and precursor synthesis. The coat protein of Bacillus subtilis spores comprises about 10% of the total dry weight of spores and 25% of the total spore protein. One protein with a molecular weight of 13,000 to 15,000 comprises a major portion of the spore coat. This mature spore coat protein has histidine at its NH2 terminus and is relatively rich in hydrophobic amino acids. Netropsin, and antibiotic which binds to A-T-rich regions of DNA and inhibits sporulation, but not growth, decreased the synthesis of this spore coat protein by 75%. A precursor spore coat protein with a molecular weight of 25,000 is made initially at t1 of sporulation and is converted to the mature spore coat protein with a molecular weight of 13,500 at t2 - t3. These data indicate that the spore coat protein gene is expressed very early in sporulation prior to the modifications of RNA polymerase which have been noted.


Preparation of glycopeptides and oligosaccharides from thyroxine-binding globulin. Over 99% of thyroxine (T4), the major form of thyroid hormone in plasma, is bound to the plasma glycoprotein thyroxine-binding globulin (TBG). The carbohydrate composition of TBG (14.6% by weight) consists of mannose, galactose, N-acetylglucosamine, and N-acetylneuraminic acid in the molar ratios of 11:9:16:10 per mol of glycoprotein. No fucose or N-acetylgalactosamine were detected. Amino acid analyses were performed. Glycopeptides, prepared by exhaustive pronase treatment of the glycoprotein, were separated by gel filtration and ion exchange chromatography. All glycopeptides contained the four sugars present in the native glycoprotein. One-fourth of the glycopeptide fraction was resolved into a discrete component, glycopeptide I. The remaining glycopeptides were a mixture termed glycopeptides II and III. Glycopeptides II and III were resolved into two discrete carbohydrate units, termed oligosaccharides A and B, by alkaline-borohydride treatment and DEAE-cellulose chromatography. We propose that TBG contains four oligosaccharide chains as calculated from the molecular weights of the glycopeptides and from compositional data assuming 1 asparagine residue/glycopeptide. The carbohydrate structures of the glycopeptides and relative affinities of TBG, glycopeptides and oligosaccharides for hepatocyte plasma membrane binding are presented in the accompanying paper (Zinn, A.B., Marshall, J.S., and Carlson, D.M. (1978) J. Biol. Chem. 253, 6768-6773.


Fluctuations in the production of specific cellular peptides during the growth of animal cells. Patterns of newly synthesized proteins of Vero cells in different growth states were obtained using two-dimensional gel electrophoresis. The 240 most prevalent peptide spots were then compared. Cells in exponential growth and in the stationary phase were found to have patterns of peptide spots characteristic of their state of growth. The transition between these patterns is progressive, and the cells acquire a pattern characteristic of quiescent cells by the late exponential phase. These observations suggest that a series of modulations in gene expression occurs during the transition of growth states in animal cells that leads to the specific appearance or disappearance of certain cellular peptides.


Outer membrane protein antigens in an enzyme-linked immunosorbent assay for Salmonella enteric fever and meningococcal meningitis. Outer membrane protein preparations were obtained from strains of Salmonella and Neisseria meningitidis. Solubilized cell envelope (CE) fractions from S. typhi and Salmonella groups A, B, C, and E had very similar electrophoretic mobilities on polyacrylamide gel, and common antigens were demonstrated by immunodiffusion. CE appeared to be a more satisfactory antigen than the more purified preparation (T/TEI) in the enzyme-linked immunosorbent assay (ELISA) with sera from typhoid and paratyphoid patients. With either antigen, however, the presence of antibodies was demonstrated in acute- and vonvalescent-phase sera. In the case of N. meningitidis infections, the crude (STA) and the more purified antigens (T/TEI) were equally satisfactory, and a rise in antibody titer could easily be demonstrated with paired acute- and convalescent-phase sera. The ELISA appears to be a simple but highly sensitive test for the detection of antibodies by using outer membrane protein antigens.


Piliation and colonial morphology among laboratory strains of meningococci. Colonial morphology and piliation were studied on twelve strains from various serogroups of Neisseria meningitidis. Six different colony types (M1 to M6) were identified. Most strains elaborated only an M1 colonial type, which is similar to gonococcus T4. Several combinations of piliation and colonial morphology were observed: (i) colonial variation in which neither parent nor variant were piliated; (ii) colonial variation involving piliated and nonpiliated cells; (iii) dissociation of piliated from nonpiliated cells with no colonial change; and (iv) colonial variation in which both variants were piliated but with distinctly different pili. Results of this study demonstrate that correlations between piliation and colony morphology within N. meningitidis are exceptions rather than the rule.


Plaque formation by strains of spotted fever rickettsiae in monolayer cultures of various cell types. Some strains of spotted fever rickettsiae could be distinguished by their ability or inability to form plaques in monolayer cultures of various mammalian and avian cell types.


The effect of acetazolamide on cerebral blood flow and oxygen utilization in the rhesus monkey. The brain is critically dependent for its moment to moment function and survival on an adequate supply of oxygen. The enzyme carbonic anhydrase (EC 4.2.1.1) may play an important role in oxygen delivery to brain tissue by facilitating the hydration of metabolically produced carbon dioxide in erythrocytes in brain capillaries, thus permitting the Bohr effect to occur. We examined the effect of 30 mg/kg i.v. acetazolamide, a potent inhibitor of carbonic anhydrase, upon cerebral blood flow and oxygen consumption in lightly anesthetized, passively ventilated rhesus monkeys. Cerebral blood flow and oxygen consumption were measured with oxygen-15-labeled water and oxygen-15-labeled oxyhemoglobin, respectively, injected into the internal carotid artery and monitored externally. Acetazolamide produced an immediate and significant increase in cerebral blood flow (from a mean of 64.7 to 83.8 ml/100 g per min), an increase in arterial carbon dioxide tension (from a mean of 40.7 to 47.5 torr), and a decrease in cerebral oxygen consumption (from a mean of 4.16 to 2.82 ml/100 g per min). Because the change in cerebral oxygen consumption occurred within minutes of the administration of acetazolamide, we believe that this effect probably was not due to a direct action on brain cells but was achieved by an interference with oxygen unloading in brain capillaries. A resultant tissue hypoxia might well explain part of the observed increase in cerebral blood flow.


Inactive form of erythrocyte carbonic anhydrase B in patients with primary renal tubular acidosis. Evidence was found for an inactive form of carbonic anhydrase type B in the erythrocytes of two children with primary renal tubular acidosis. The addition of zinc chloride to hemolysates from these patients resulted in a marked increase in the activity of this enzyme. No such effect was noted with hemolysates of control subjects. No significant differences were observed in the zinc levels of hemolysates of these patients and of normal individuals. However, the level of zinc in the carbonic anhydrase B isolated from one of these patients was low, suggesting a modified form of the enzyme. The restoration of activity upon the addition of zinc was reversed by ethylenediamine tetraacetate, but no such effects were noted for the carbonic anhydrase B of normal individuals. Thus the abnormal carbonic anhydrase B has decreased zinc binding. The ultraviolet difference spectrum of the carbonic anhydrase B of normal individuals and that of a patient showed a peak at 305 nm which decreased upon the addition of zinc. The abnormal form of carbonic anhydrase B was not distinguishable from that of normal individuals by either immunological or electrophoretic criteria.


Evaluation of some methods for the laboratory examination of sputum. Methods for the quantitation of leucocytes, squamous epithelial cells, and potential pathogens in sputa are described. Microscopic examination showed that 58% of sputa tested (554/957) were purulent or moderately purulent and 48% were moderately or heavily contaminated by squamous epithelial cells. The presence of squamous epithelial cells indicated oropharyngeal contamination. A simple dilution technique was chosen to compare the isolation of potential pathogens from direct cultures and from dilutions of sputa (10(-7) per ml original sputum). The dilution technique permitted easier reading of sputum cultures and avoided the possible over-reporting of enterobacteria and Pseudomonas aeruginosa, which were frequently found on direct examination. Enterobacteria and Pseudomonas aeruginosa were more likely to be isolated from sputa moderately or heavily contaminated with squamous epithelial cells. The use of counter-current immunoelectrophoresis (CIE) and co-agglutination tests to detect pneumococcal antigen in sputa is described. The presence of antigen in sputum was a more reliable index of lower respiratory tract infection than a positive culture. Co-agglutination tests were simpler to perform and used smaller amounts of expensive antiserum than CIE.


Intracortical connectivity of architectonic fields in the somatic sensory, motor and parietal cortex of monkeys. Anterograde and retrograde transport methods were used to study the corticocortical connectivity of areas 3a, 3b, 1, 2, 5, 4 and 6 of the monkey cerebral cortex. Fields were identified by cytoarchitectonic features and by thalamic connectivity in the same brains. Area 3a was identified by first recording a short latency group I afferent evoked potential. Attempts were made to analyze the data in terms of: (1) routes whereby somatic sensory input might influence the performance of motor cortex neurons; (2) possible multiple representations of the body surface in the component fields of the first somatic sensory area (SI). Apart from vertical interlaminar connections, two types of intracortical connectivity are recognized. The first, regarded as "non-specific," consists of axons spreading out in layers I, III and V-VI from all sides of an injection of isotope; these cross architectonic borders indiscrimininately. They are not unique to the regions studied. The second is formed by axons entering the white matter and re-entering other fields. In these, they terminate in layers I-IV in one or more mediolaterally oriented strips of fairly constant width (0.5--1 mm) and separated by gaps of comparable size. Though there is a broadly systematic topography in these projections, the strips are probably best regarded as representing some feature other than receptive field position. Separate representations are nevertheless implied in area 3b, in areas 1 and 2 (together), in areas 3a and 4 (together) and in area 5; with, in each case, the representations of the digits pointed at the central sulcus. Area 3b is not connected with areas 3a or 4, but projects to a combined areas 1 and 2. Area 1 is reciprocally connected with area 3a and area 2 reciprocally with area 4. The connectivity of area 3a, as conventionally identified, is such that it is probably best regarded not as an entity, but as a part of area 4. Areas identified by others as area 3a should probably be regraded as parts of area 3b. Parts of area 5 that should be more properly considered as area 2, and other parts that receive thalamic input not from the ventrobasal complex but from the lateral nuclear complex and anterior pulvinar, are also interconnected with area 4. More posterior parts of area 5 are connected with laterally placed parts of area 6. A more medial part of area 6, the supplementary motor area, occupies a pivotal position in the sensory-motor cortex, for it receives fibers from areas 3a, 4, 1, 2 and 5 (all parts), and projects back to areas 3a, 4 and 5.


The hypothalamic magnocellular system of the rhesus monkey: an immunocytochemical study. The hypothalamic magnocellular system of the adult female rhesus monkey is studied here, using immunoperoxidase technique and antisera to estrogen stimulated neurophysin (ESN), nicotine stimulated neurophysin (NSN), oxytocin (OT) and vasopressin (VP). These observations confirm and enlarge previous descriptions by others using Gomori techniques. It is apparent from this study that the magnocellular system spreads through a broader area than is generally accepted. A group of cells ventral to the head of the caudate nucleus and medial to the internal capsule is described. The general orientation of the nuclei and their tracts can only be appreciated when coronal, horizontal and sagittal sections are compared. Our observations suggest that the supraoptic nucleus is made up of a single group of cells that straddles the optic pathways, and is not divided in three segments, as it is generally described. It is also shown that the rostral extensions of the paraventricular and supraoptic nuclei merge above the optic chiasm. Cells containing ESN/OT and NSN/VP are evenly dispersed in the paraventricular nucleus but a topographical arrangement is present in the supraoptic nucleus. The magnocellular nuclei project to the organum vasculosum of the lamina terminalis, zona externa of the median eminence and pars nervosa of the pituitary gland. Reactive fibers were also seen within islets of cells from the pars intermedia located inside the pars nervosa. A globular structure containing small blood vessels surrounded by positive fibers was noticed protruding into the floor of the third ventricle, at the level of the median eminence.


A comparative study of the neurons of origin of the spinocerebellar afferents in the rat, cat and squirrel monkey based on the retrograde transport of horseradish peroxidase. The cell bodies of the neurons of the spinocerebellar pathways were examined using large injections of horseradish peroxidose into the cerebellum. Sections of each spinal segment were examined with both the DAB and the de Olmos O-dianisidine techniques. Results common to all three species were found. In Clarke's nucleus, the central cervical nucleus, and the spinal border cells there were many heavily labeled cells. Clarke's nucleus was found to project primarily ipsilaterally; the spinal border cells primarily contralaterally; and the central cervical nucleus bilaterally. In addition to these aggregates of spinocerebellar neurons there were numerous labeled neurons scattered throughout the spinal grey. Labeled neurons were found in all portions of the spinal grey except the substantia gelatinosa and lateral cervical nucleus and occurred in all spinal segments. They varied in morphology from large multipolar neurons, found predominantly in the ventral horn to small globular and fusiform neurons that were most abundant in the dorsal horn. These cells were found to project both ipsilaterally and contralaterally. Results common to only two of the species examined were also found. In the dquirrel monkey and the cat, but not the rat, the marginal layer of the dorsal horn in all segments of the spinal contained numerous labeled neurons. These marginal neurons were especially numerous in the squirrel monkey, where as many as 13 to 16 labeled neurons per section of the dorsal horn were found. In the rat and the squirrel monkey but not the cat, some intensely labeled large multipolar neurons were found in the sacral and caudal segments. These are the cells of Stilling's nucleus, a column of cells similar in position and orientation to that of Clarke's column but different in its projections and details of cytoarchitecture. Thus we have not only confirmed that Clarke's nucleus, the central cervical nucleus, and the spinal border cells project to the cerebellum but we have also found several new sources of spinocerebellar afferents.


Aflatoxin production on high moisture corn and sorghum with a limited incubation. Samples of ground, cracked, or whole kernels of sorghum or corn were inoculated with Aspergillus flavus no. 15546. The samples were incubated at 25 or 30 C and 90% relative humidity for 48 or 72 h. In all treatments, the 72-h samples contained more aflatoxins B1 and B2 than the 48-h samples. Cracked sorghum at 30  C for 72 h and whole sorghum at 25 C for 72 h contained more aflatoxins than any other treatment. Even in these short incubation periods, enough aflatoxin could be produced to be harmful to livestock.


Serum thyrotropin, thyroxine, and tri-iodothyronine in dairy cows fed varying amounts of iodine. The objective was to determine whether dairy cows fed supplemental iodine daily at 2.5 or 5.0 mg per kg body weight for 49 wk had altered thyroid status relative to controls fed no supplemental iodine. Average daily dose of iodine was 1.6 and 3.3 g. Iodine as ethylene diamine dihydriodide was fed to Holstein cows beginning at 8 to 10 wk of lactation, for the remainder of that lactation, through the dry period, and into the next lactation. On day 341 after onset of iodine feeding, each cow was injected intravenously with 15 microgram thyrotropin releasing hormone per 100 kg body weight. Thyrotropin in jugular blood collected before injection of releasing hormone averaged 7.6, 6.1, and 8.2 ng/ml for cows given 0, 2.5, and 5.0 mg iodine/kg body weight; there were no differences between means. Releasing hormone increased thyrotropin, thyroxine, and tri-iodothyronine in serum of all cows, but increases were not affected by quantity of dietary iodine.


Surgical repair of lateral ankle instability by modification of traditional approaches. Recurrent instability following ankle sprains which fail to respond adequately to conservative treatment has been the focal point of discussion among physicians in recent years. A variety of surgical regimens have been implemented primarily over the past 40 years in an attempt to control this problem. The authors discuss these modalities and present their successful approach in treating lateral ankle instability.


Acetylation of chromosome squashes of Drosophila melanogaster decreases the background in autoradiographs from hybridization with [125I]-labeled RNA. DNA in prepared chromosomes from the larval salivary glands of Drosophila melanogaster was hybridized with [125I]-labeled 5S and tRNA from the same organism. Autoradiography revealed that radioactivity was frequently bound to all regions of the slides, masking labeling of the chromosomes. Acetylation of the preparations before hybridization prevented the formation of this background and revealed the specific chromosomal sites.


Stress and menstrual distress.

Stress and menstrual distress. The relationship between stress and menstrual distress is considered in response to three questions: (1) How stressful are menstruation and its associated events? (2) What factors serve to make menstruation stressful? (3) What are the effects of stress upon the menstrual cycle? Included in the analysis are estimates of the incidence of negative symptoms associated with the menstrual cycle, other indicators of the stressfulness of the premenstrual and menstrual phases, adrenal factors, animal studies, and suggested directions for research and practice.


Waterborne infectious disease in Britain. The incidence of waterborne typhoid fever and other intestinal infections in Britain since 1937 is reviewed. In the light of the current very low incidence of indigenous typhoid fever, it is suggested that the examination of relevant waterworks staff and prospective employees for carriage of the agents of potentially waterborne disease should be done selectively by appropriate laboratory tests only when indications for them are revealed by medical assessment.


The relative effectiveness of commonly used disinfectants in inactivation of echovirus 11. Echovirus 11 in the presence of fetal calf serum was exposed to six commonly used disinfectants for times of 10, 20 and 30 s. At the end of such exposure times, skim milk neutralized disinfectant activity and residual virus was assayed using the plaque technique. The six disinfectants studied were Javex, sodium hydroxide, ethanol, Wescodyne, One Stroke Ves-Phene, and Sonacide. Although 0.25% (w/v) sodium hydroxide and 95% (v/v) ethanol were equally virucidal and significantly more so than the other four disinfectants, causing 10(6) reduction in 20 s, they may not be practical to use in many instances. Javex at a dilution of 1/50 (1200 parts/10(6) chlorine) proved to be virucidal causing 10(3.5) reduction of echovirus 11 in 30 s. Wescodyne (1/50) and undiluted Sonacide were relatively ineffective causing 10 reduction or less of echovirus 11 in 30 s. One Stroke Ves-Phene (1/50) was ineffective causing no significant inactivation in 30 s.


Studies on the transfer of protective immunity with lymphoid cells from mice immune to malaria sporozoites. In an effort to understand the mechanisms involved in the protective immunity to malarial sporozoites, an A/J mouse/Plasmodium berghei model was studied. Protective immunity could consistently be adoptively transferred only by using sublethal irradiation of recipients (500 R); a spleen equivalent (100 X 10(6))of donor cells from immune syngeneic mice; and a small booster immunization (1 X 10(4)) of recipients with irradiation-attenuated sporozoites. Recipient animals treated in this manner were protected from lethal challenge with 1 X 10(4) nonattenuated sporozoites. Immune and nonimmune serum and spleen cells from nonimmune animals did not protect recipient mice. Fewer immune spleen cells (50 X 10(6)) protected some recipients. In vitro treatment of immune spleen cells with anti-theta sera and complement abolished their ability to transfer protection. This preliminary study suggests that protective sporozoite immunity can be transferred with cells, and that it is T cell dependent.


Antibodies to guinea pig lymphokines. VII. Reactivity with products of lymphoid and nonlymphoid cells. It was shown previously, that an antiserum directed against highly purified fractions of migration inhibitory factor inhibits delayed hypersensitivity reactions in vivo and in vitro. Using radiolabeling techniques we determined that the anti-lymphokine serum reacted primarily with three lymphocyte activation products (m.w. 60,000, 45,000, and 30,000) all of which had a similar isoelectric point of 5.2. The cellular origin of this material was investigated. It was found that activated B cells, B leukemia cells (L2C), and growing fibroblasts produced material of a similar m.w. as analyzed on SDS-PAGE. No cross-reaction was found with radiolabeled products of activated murine and human lymph node cells and of SV 40-infected African green monkey kidney cells. The isoelectric point of the reactive material from B cells, leukemia cells, and fibroblasts was determined at 5.2. In addition to material with pI 5.2, lymph node cells also produced material with pI 3.5 to 4.5, which focused at pH 5.0 to 5.4. After neuraminidase treatment macrophage migration inhibitory activity in fibroblast culture supernatants could be absorbed specifically to insolubilized anti-lymphokine antibody. These findings suggests that lymphoid and nonlymphoid cells are capable of producing molecules whose physicochemical and functional properties appear to be identical.


Inhibition of migration of tumor cells in vitro by lymphokine-containing supernatants. Lymphokine-containing supernatants derived from seven different human lymphoid cell lines and lymphokine-containing supernatants from concanavalin A-stimulated murine lymphocytes were found to be capable of reversibly inhibiting the migration of tumor cells in vitro. The tumor cell lines used in these studies were the P815 mastocytoma, Ehrlich ascites, Walker carcinosarcoma, Hepatoma 129, and Sarcoma 37. Preliminary physiochemical evidence suggests that the mediator, here termed TMIF, is distinct from MIF. In any case, these results suggest the possibility that lymphokines other than lymphotoxin or macrophage-activating factors may play a role in tumor immunity.


Genetic control of immune response to sperm whale myoglobin in mice. I. T lymphocyte proliferative response under H-2-linked Ir gene control. Studies on the genetic control of immune response to sperm whale myoglobin were initiated. As demonstrated in this paper, the T lymphocyte proliferative response to whale myoglobin is under H-2-linked Ir gene control. Mice of H-2d, H-2f, and H-2s haplotypes were high responders to the myoglobin, whereas haplotypes H-2b, H-2k, H-2p, H-2q, and H-2r were low responders. The Ir gene(s) was localized between H-2K and H2D regions, since the recombinant strain A.TL (KsIkSkDd) was a low responder and A.TH (KsIsSsDd) was a high responder. Further studies with recombinant strains revealed that the expression of the high-responder I-Ad or Ias alleles was sufficient to give a good response, since strains D2.GD (d d b b b b b b) and B10.HTT (s s s s k k k d) were high responders. The expression of the I-Cd allele in strains B10.A (k k k k k d d d) and B10.A(5R) (b b b k k d d d) also gave high response, and thus suggested a second Ir gene, derived from the H-2d haplotype. The finding that expression of the I-Cs allele in B10.S(8R) (k k ? ? s s s s) did not result in high response suggests the lack of the second Ir gene in the high-responder H-2s haplotype.


Structural studies on induced antibodies with defined idiotypic specificities. VI. Amino terminal sequences of the heavy and light chain variable regions of anti-p-azophenylarsonate antibodies from A/J mice suppressed for a cross-reactive idiotype. Previous studies in this series have been directed toward the elucidation of the heavy and light chain variable region structures of antibodies raised in A/J mice to the p-azophenylarsonate (Ar) hapten, certain of which bear a cross-reacting idiotype. The present study concerns an analysis of anti-Ar antibodies that arise in A/J mice suppresed for a cross-reacting idiotype. The results indicate that when an idiotype is suppressed and the animal subsequently hyperimmunized, the resultant antibodies are "deviated" into different V-region subgroups, both in the heavy and light polypeptide chains. The study presents the first primary structural analysis of a humoral immune response that has been manipulated by an idiotypic reagent.


Estimation of antibodies specific for dextran. Methods are described for the isolation and characterization of picogram quantities of anti-dextran antibodies. 14C-dextrans produced by using the dextransucrases of Leuconostoc mesenteroides strains B1355 and B512 were used in a radioimmunoassay. The specificity of this assay was verified by using cell cytoplasmic lysates from mouse plasmacytomas, J558 (anti-alpha 1 leads to 3 dextran) and W3129 (anti-alpha 1 leads to 6 dextran). Dextran produced by strain B1355 and insolubilized with epichlorohydrin was used as an immunoabsorbent.


Identification of tyrosinase inhibitors in cultures of Pityrosporum. Lipid fractions capable of inhibiting the dopa-tyrosinase reaction in vitro were isolated by thin-layer chromatography from submerged cultures of Pityrosporum supplemented with oleic acid or vaccenic acid. Analysis of these fractions by gas chromatography-mass spectrometry revealed the presence mainly of C(9) and C(11) dicarboxylic acids. Standard dicarboxylic acids from C(8) to C(13) were capable of inhibiting tyrosinase in vitro to varying extents. Enzymatic kinetic studies showed that they act as competitive inhibitors of tyrosinase. These observations suggest that dicarboxylic acids could be used in the treatment of people with hyperpigmentary disorders.