[Vitreo-corneal adhesions. A clinico-anatomical and ultrastructural study (author's transl].

Study of six cases of vitreous touch syndrom with histopathology of five buttoms of kératoplasty. Light and electron-microscopy demonstrate many alterations of Descemet's membrane, which is thickened, devided in sheets. In one specimen there was a retrocorneal fibrous membrane and "fibroblast like cells". These findings support the mechanism of endothelial fibrous metaplasia as the origin of the retrocorneal fibrous membrane during vitreous touch syndrom.


[Treatment of alternating squints: method of localisation by prismatic hypercorrection and with penalisation (author's transl)].

An analysis of the results of treatment of 50 cases of alternating squint by prismatic hypercorrection with penalisation. The authors have considered the decrease in the deviation, the creation of binocular vision and the durability of the effects after one year. These results equally take into account the age at which the squint appeared.


[The lashes. Physiology, structure, pathology (author's transl)].

In spite of the similarity between hairs and eyelashes the later shows several physiological and anatomical particularities. We performed an ultrastructural study of eyelashes and reviewed their pathology.


[Aetiological aspects of uveitis (author's transl)].

An analysis of the probable aetiology of 222 cases of uveitis. In order of frequency the aetiology was: infectious focis 31%, rheumatism 28%, tuberculosis 7.6%, streptococci 4%, viral 3.6%, rickettsia 1.3%, syphilis, brucellosis in 0.5%. In 22.1% of cases the aetiology was not established. In young subjects the majority consisted of bacterial and viral infections particularly streptococcal. In adults the most frequent aetiology was rheumatic and in the elderly foci of infection.


[François syndrome: two observations (author's transl)].

Two cases of "Francois Syndrome" were observed at the C.H.U. in Rabat. All the signs defined by Francois himself were found in these two cases which also demonstrated certain aspects which raised the aetiopathpological problem of the condition (the hereditary of embryopathic theory).


Osmotic adaptation to rigid contact lenses.

Using a precision thermocouple hygrometer, requiring just 5 microliters of sample, the tear osmotic pressures of 6 subjects were monitored throughout their adaptation to rigid contact lenses. Relative to the prefitting baselines established for each previous to this study, these subjects showed a range of hypotonic shifts on initially receiving their lenses, but this imbalance was rectified by the end of the first wearing week and a new relatively hypertonic level of tear osmotic pressure was commonly established. This new baseline was reflected in the pre-insertion samples of several of the subjects as well once adaptation was advanced.


[The action of instilled physostigmine on the acetylcholine content of the bovine retina].

Kymographic measurements of acetylcholine (ACh) in bovine retinal tissue carried out on guinea-pig ileum do not indicate a significant elevated concentration of ACh in the eye instilled with physostingmine salicylate (1% aqueous solution, 10 applications of three drops at intervals of 3 minutes). The eyes were enucleated one hour after the last application.


Effect of some Pteridine compounds on the Na+ + K+)-ATPase and on the cardiac glycoside receptor of human heart).

Pteridine compounds are known to block Na+-reabsorption and K+-secretion in epithelial cells (salivary duct of the rat), which actively transport Na+ and K+ against an electrochemical gradient. Furthermore, there have been reports on antagonistic effects of these substances in digitalis induced arrhythmias. Therefore the actions of triamterene (Jatropur, Dyrenium), the sulfuric acid ester and the methylether of p-hydroxytriamterene (OH-triamterene) and OH-triamterene on specific [3H] g-strophanthin (ouabain) binding and Na+ + K+)-ATPase activity of isolated human cardiac cell membranes were investigated.  Triamterene, the sulfuric acid ester and the methylether of OH-triamterene inhibit (Na+ + K+)-ATPase activity only at very high concentrations (10(-5)--10(-4) M). OH-Triamterene does not inhibit this enzyme at concentrations lower than 10(-3) M. The specific binding of [3H] g-strophanthin to human cardiac cell membranes is inhibited half maximally at relatively high concentrations, too (10(-5)--10(-4) M). These results are rather indicative of unspecific effects due to membrane sites of action other than the (Na+ + K+)-ATPase or the cardiac glycoside receptor.


Electrocardiographic criteria of left ventricular hypertrophy in left bundle-branch block.

In order to determine whether the electrocardiographic criteria of left ventricular hypertrophy apply in the presence of left bundle-branch block we studied 79 cases of intermittent left bundle-branch block and compared the QRS voltage and axis before and after its onset. Cases of incomplete left bundle-branch block were excluded. There was a statistically significant correlation between pre- and post-left bundle-branch block values of R or S wave voltage in leads I, V1, V2, V5, and V6, the Sokolow index (R V5 or V6 + S V1), and the QRS axis. There was a statistically significant reduction in R wave voltage in leads I, V5, and V6, an increase in S wave voltage in V1 and V2, and leftward shift of QRS axis, but the Sokolow index remained unchanged, after the onset of left bundle-branch block. The Sokolow criteria for left ventricular hypertrophy apply satisfactorily even in the presence of left bundle-branch block, though specificity is low, but QRS axis is unhelpful.


Pressure-volume relationships in the lung during laparoscopy.

Pressure-volume relationships for the total respiratory system and for the lung were recorded in anaesthetized and paralysed patients, during deflation from an airway pressure of 3 kPa to FRC at a rate of 2 litre min-1. Pleural pressure was estimated by means of an oesophageal balloon. A group of nine female patients (mean age 32.7 yr) about to undergo laparoscopy were each studied in four successive states: supine, 15 degrees head down tilt, tilt and lithotomy position, and again in this position after abdominal inflation with nitrous oxide to a pressure of 0.8(-1) kPa. Compliance values were calculated from the curves. Mean total compliance was increased significantly by moving to the lithotomy position, and reduced markedly after inflation of the abdomen, because of a large reduction in thoracic compliance. Mean lung compliance was unaltered, except for a slight but statistically significant increase on moving from the supine to the Trendelenburg position. Measurement of FRC by helium dilution in a group of seven patients showed that abdominal inflation caused a mean decrease of 19%. Airway closure manoeuvres were carried out using a helium bolus technique from FRC in five patients, but closing volume could be measured in only one patient, in the supine position. The absence of an inflexion in the slope of the pressure-volume curves for the other patients supported this negative finding.


HLA C and D antigens associated with psoriasis.

The frequency of two newly defined HLA antigens, HLA CT7 and DMA, was found to be greatly increased in patients with psoriasis. These two antigens and those previously found to be associated with the disease, B13, BW16 and BW17, frequently occurred together. The disease may be primarily associated with HLA DMA, or with the HLA haplotypes CT7--B13/W16/W17--DMA.


Occupational allergic contact dermatitis due to leatherleaf fern Arachniodes adiantiformis (Forst) Tindale.

Leatherleaf fern has been imported from the United States, Honduras and Puerto Rico to the Federal Republic of Germany since 1966 and cultivated in Germany since 1970/71. It is slowly displacing the floral arrangements with asparagus used until now as filler and background material. This change is due primarily to its better keeping quality. Continuous contact with the leatherleaf fern induced an allergic contact dermatitis of the palms and fingers in a female florist. Patch tests with the leaf revealed the allergic origin of the dermatitis. Five fractions of a chromatographically separated extract of the fern were tested on sensitized guinea-pigs and on the patient. Only the first fraction gave a positive patch test response. By further separation colourless crystals of the contact allergen could be obtained. Elucidation of the structure is in progress. The sensitizer could only be detected during sporogenesis of the fern, that means when sporecapsules are present on the surface of the leaves.


Evidence for a catalytic function of the coupling factor 1 protein reconstituted with chloroplast thylakoid membranes.

The effects of tentoxin on the ATPase activities of coupling factor 1 proteins (CF1) and photophosphorylation with isolated chloroplasts and chloroplasts reconstituted with coupling factor proteins have been examined. 1. The calcium-dependent ATPase activities of coupling factors isolated from spinach, lettuce and Nicotiana otophora are completely inhibited by tentoxin. The ATPase activities of coupling factors isolated from Nicotiana tabacum and Nicotiana knightiana are not affected by tentoxin. 2. Phenazine methosulfate-catalyzed cyclic photophosphorylation with chloroplasts isolated from spinach, lettuce and N. otophora is completely inhibited by tentoxin, whereas chloroplasts isolated from N. knightiana and N. tabacum are relatively insensitive to tentoxin. 3. Spinach chloroplasts, partially depleted in CF1, can be reconstituted with coupling factors isolated from a wide variety of plants including lettuce, radish, N. tabacum, N. knightiana and N. otophora. 4. Spinach chloroplasts reconstituted with spinach, lettuce and N. otophora CF1 retain their sensitivity to tentoxin; however, when reconstituted with N. knightiana and N. tabacum coupling factor proteins, a significant fraction of the reconstituted rate remains tentoxin insensitive. These data are interpreted as evidence that coupling factors that reconstitute with spinach thylakoid membranes have both a catalytic and structural function.


The initial stages of photophosphorylation. Studies using excitation by saturating, short flashes of light.

1. Photophosphorylation was studied in spinach chloroplasts on illumination, from the dark state, with saturating short ("single turnover") flashes of light. 2. At rapid flash rates (100 Hz), phosphorylation began within the first five flashes. The ATPase inhibitor protein appeared to be displaced from its inhibitory site on the ATPase also within five flashes, as deduced from the flash-induced ATPase activity. 3. At slower flash rates, or if the rate of electron transfer were reduced with 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU), phosphorylation began only after a larger number (50--60) of flashes. The displacement of the ATPase inhibitior protein was similarly delayed. 4. Partial displacement of the inhibitor protein from its inhibitory site on the ATPase (by pretreatment with dithioerythritol) allowed phosphorylation to proceed without a perceptible lag, even in the presence of DCMU. It was concluded that the ATPase inhibitor protein must be displaced on the ATPase before phosphorylation can begin, and that this process is energy dependent. 5. During the flash regime used, release of inhibitor from its inhibitory site seemed to be governed largely by the membrane potential. The light-induced pH gradient seemed to have little effect under these conditions. Our results are not compatible with a direct conformational interaction between the electron transfer chain and the ATPase causing displacement of the inhibitor. 6. The maximal rate of photophosphorylation induced by less than 200 flashes was 0.12--0.15 mol ATP made/mol ATPase per flash. This rate seemed to be limited not be the supply of energy to the ATPase molecules, nor by the maximal turnover capacity of the ATP synthesising system, but by the number of ATPase molecules which were active in synthesis, i.e., which lacked the inhibitor protein. 7. The bound nucleotides of the coupling ATPase exchanged with added nucleotides during single turnover flashes. At high flash rates, exchange began within 5 flashes. The average amount of nucleotide exchanged per flash over 100 flashes was about one tenth the amount of ATP synthesised in each flash. 8. It is concluded that, during phosphorylation, a steady state level of active coupling ATPases is set up. The energy-dependent displacement of the inhibitor protein, and its (energy-independent) relaxation back on to the inhibitory site are the two opposing factors involved in this steady state.


Membrane (Na+ + K+)-ATPase of canine brain, heart and kidney. Tissue-dependent differences in kinetic properties and the influence of purification procedures.

Effects of commonly used purification procedures on the yield and specific activity of (Na+ + K+)-ATPase (Mg2+-dependent, Na+ + K+-activated ATP phosphohydrolase, EC 3.6.1.3), the turnover number of the enzyme, and the kinetic parameters for the ATP-dependent ouabain-enzyme interaction were compared in canine brain, heart and kidney. Kinetic parameters were estimated using a graphical analysis of non-steady state kinetics. The protein recovery and the degree of increase in specific activity of (Na+ + K+)-ATPase and the ratio between (Na+ + K+)-ATPase and Mg2+-ATPase activities during the successive treatments with deoxycholate, sodium iodide and glycerol were dependent on the source of the enzyme. A method which yields highly active (Na+ + K+)-ATPase preparations from the cardiac tissue was not suitable for obtaining highly active enzyme preparations from other tissues. Apparent turnover numbers of the brain (Na+ + K+)-ATPase preparations were not significantly affected by the sodium iodide treatment, but markedly decreased by deoxycholate or glycerol treatments. Similar glycerol treatment, however, failed to affect the apparent turnover number of cardiac enzymes preparations. Cerebral and cardiac enzyme preparations obtained by deoxycholate, sodium iodide and glycerol treatments had lower affinity for ouabain than renal enzyme preparations, primarily due to higher dissociation rate constants for the ouabain.enzyme complex. This tissue-dependent difference in ouabain sensitivity seems to be an artifact of the purification procedure, since less purified cerebral or cardiac preparations had lower dissociation rate constants. Changes in apparent association rate constants were minimal during the purfication procedure. These results indicate that the presentyl used purification procedures may alter the properties of membrane (Na+ + K+)-ATPase and affect the interaction between cardiac glycosides and the enzyme. The effect of a given treatment depends on the source of the enzyme. For the in vitro studies involving purified (Na+ + K+)-ATPase preparations, the influence of the methods used to obtain the enzyme preparation should be carefully evaluated.


Role of negatively charged phospholipids in highly purified (Na+ + K+)-ATPase from rabbit kidney outer medulla studies on (Na+ + K+)-activated ATPase, XXXIX.

1. The requirement for specific polar head groups of phospholipids for activity of purified (Na+ + K+)ATPase from rabbit kidney outer medulla has been investigated. 2. Comparison of content and composition of phospholipids in microsomes and the purified enzyme indicates that purification leads to an increase in the phospholipid/protein ratio and in phosphatidylserine content. 3. The purified preparation contains 267 molecules phospholipid per molecule (Na+ + K+)-ATPase, viz. 95 phosphatidylcholine, 74 phosphatidylethanolamine, 48 spingomyelin, 35 phosphatidylserine and 15 phosphatidylinositol. 4. Complete conversion of phosphatidylserine into phosphatidylethanolamine by the enzyme phosphatidylserine decarboxylase has no effect on the (Na+ + K+)-ATPase activity of the purified preparation. 5. Complete hydrolysis of phosphatidylinositol by a phospholipase C from Staphylococcus aureus, which is specific for this phospholipid, has no effect on the (Na+ + K+)-ATPase activity. 6. Hydrolysis of 95% of the phosphatidylcholine and 60--70% of the spingomyelin and phosphatidylethanolamine by another phospholipase C (Clostridium welchii) lowers the (Na+ + K+)-ATPase activity by about 20%. 7. Combination of the phospholipid-converting enzymes has the same effect as can be calculated from the effects of the enzymes separately. Only complete conversion of both phosphatidylserine and phosphatidylinositol results in a loss of 44% of the (NA+ + K+)-ATPase activity and 36% of the potassium 4-nitrophenylphosphatase activity. 8. These experiments indicate that there is no absolute requirement for one of the polar head groups, although in the absence of negative charges the activity is lower than in their presence.


Phosphatidylinositol as the endogenous activator of the (Na+ + K+)-ATPase in microsomes of rabbit kidney.

Incubation of rabbit kidney microsomes with pig pancreatic phospholipase A2 produced residual membrane preparations with very low (Na+ + K+)-ATPase activity. The activity could be restored by recombination with lipid vesicles of negatively-charged glycerophospholipids. Vesicles of pure phosphatidylcholine and phosphatidylethanolamine were virtually inactive in this respect, but could reactivate in the presence of cholate. Incubation of the microsomes with a combination of phospholipase C (Bacillus cereus) and spingomyelinase C (Staphylococcus aureus) resulted in 90--95% release of the phospholipids. The residual membrane contained only phosphatidylinositol and still showed 50--100% of the (Na+ + K+)-ATPase activity.


Purification from brain of an intrinsic membrane protein fraction enriched in (Na+ + K+)-ATPase.

A microsomal fraction from canine brain gray matter has been extracted with the detergent sodium dodecyl sulfate to partially purify the membrane-bound (Na+ + K+)-stimulated adenosine triphosphatase. Phospholipid, glycolipid, and a family of other glycoproteins are also enriched by the procedure; it is proposed that the product is an intrinsic membrane protein fraction. 6--8-fold purification of (Na+ + K+)-ATPase is obtained without solubilizing the enzyme and without irreversibly altering its turnover number. Final specific activities are 350--400 mumol of ATP hydrolyzed/h per mg protein. The stimulation and reversible inactivation of the (Na+ + K+)-ATPase by dodecyl sulfate were examined for information relevant to the mechanism of action of the detergent.


Is hexokinase present in the basal lateral membranes of rat kidney proximal tubular epithelial cells?

The possible presence of hexokinase in basal lateral membranes from rat kidney proximal tubules was investigated. Basal lateral membranes were obtained from homogenates of rat kidney cortex by differential centrifugation and free flow electrophoresis. They were further purified by density gradient centrifugation. Hexokinase activity was measured as the phosphorylation of D-[U14C]glucose. Throughout the purification of the membranes, the specific activity of hexokinase decreased while that of (Na+ + K+)-ATPase increased. Hexokinase activity in all fractions could be quantitatively accounted for in terms of cytosolic and mitochondrial enzyme contributions. It is concluded that there is no hexokinase activity in basal lateral membranes from rat kidney.


The turnover of lactate dehydrogenase in skeletal muscle of the mouse. Influence of fibre type and exercise regimen.

1. The influence of fibre-type and an exercise regimen on the turnover characteristics of proteins in mouse skeletal muscle has been studied. 2. Exercise induced a weight increase in most individual muscles and a decrease in lactate dehydrogenase activity which was more marked in white muscle than red. The coincident decrease in M-subunit activity was more noticeable in fast-twitch muscles than in slow-twitch types. 3. Exercise caused a considerable and consistent increase in the rate of degradation of total soluble proteins in individual muscles, with turnover being higher in muscles with a high red fibre content. 4. The response of lactate dehydrogenase to exercise varied considerably between individual muscles as indicated by the comparative rate constants for synthesis and degradation. In general, degradation was greater in muscle with a high slow-twitch or red fibre content. 5. Overall, a considerable redistribution of protein resources was observed during this physiological perturbation (exercise) with the main emphasis away from the pool of total soluble protein towards the myofibrillar constituents. Highly individualistic responses in relation to rates of synthesis and degradation were observed for the specific protein, lactate dehydrogenase, in the separate muscle of this animal.


Comparison of the (Ca2+ + Mg2+)-ATPase proteins from normal and dystrophic chicken sarcoplasmic reticulum.

The involvement of membrane protein in dystrophic chicken fragmented sarcoplasmic reticulum alterations has been examined. A purified preparation of the (Ca2+ + Mg2+)-ATPase protein from dystrophic fragmented sarcoplasmic reticulum was found to have a reduced calcium-sensitive ATPase activity and phosphoenzyme level, in agreement with alterations found in dystrophic chicken fragmented sarcoplasmic reticulum. An amino acid analysis of the ATPase preparations showed no difference in the normal and dystrophic (Ca2+ + Mg2+)-ATPase. The (Ca2+ + Mg2+)-ATPase was investigated further by isoelectric focusing and proteolytic digestion of the fragmented sarcoplasmic reticulum. Neither of these methods indicated any alteration in the composition of the dystrophic (Ca2+ + Mg2+)-ATPase. We have concluded that the alterations observed in dystrophic fragmented sarcoplasmic reticulum are not due to increased amounts of non-(Ca2+ + Mg2+)-ATPase protein, and that the normal and dystrophic (Ca2+ + Mg2+)-ATPase protein are not detectably different.


The regulation of the decay of nitrate reductase. Evidence for the existence of at least two mechanisms of decay.

There seem to be at least two different mechanisms of decay of nitrate reductase in Neurospora in vivo: one which is very sensitive to EDTA and cycloheximide, decreases with mycelial age and is not increased by an increase in temperature from 27 to 37 degrees C, the other which is relatively insensitive to EDTA and cycloheximide, increases with the age of the mycelium and with the above temperature shift.


Demonstration in vitro of two intracellular inactivators of nitrate reductase from Neurospora.

Two different inactivators of nitrate reductase have been found in cell free preparations of Neurospora. The first (Inactivator I) is very active at pH 9, is inhibited by disodium ethylene diamine tetraacetate (EDTA) and is present in all mycelia incubated under all conditions tested; the second (Inactivator II) is very active at pH 5, is repressed by ammonia or by a metabolic product of ammonia and derepressed by nitrogen starvation, cannot be derepressed by nitrogen starvation in strain nit-2, in which a number of "ammonia-represible" enzymes are permanently repressed, and is sensitive to phenyl methyl sulfonyl fluoride. Crude extracts of mycelia contain inhibitor(s) of both inactivators.


Kinetics of the oxidation of Rhus vernicifera stellacyanin by the Co(EDTA)-- ion.

A kinetic study of the oxidation of the copper(I) form of the blue copper protein stellacyanin (St(I) by Co(EDTA)-- has been performed. Observed rate constants approach a saturation limit with increasing [Co(EDTA)--] at pH 7, consistent with a mechanism involving rapid pre-equilibrium oxidant-protein complex formation followed by rate-limiting intramolecular Cu(I) to Co(III) electron transfer: Co(EDTA)-- + St(i Qp in equilibrium Co(EDTA)-- ---St(I) Co(EDTA)-- ---St(I) k2 leads to Co(EDTA)2-- ---St(II) (Qp = 149 M--1, k2 = 0.169 sec--1; 25.1 degrees, pH 7.0 mu 0.5 M (phosphate)). Activation parameters based on k2 (deltaH not equal to = 1.8 kcal/mol, deltaS not equal to = --56 cal/mol-deg) indicate that the electron transfer process is substantially nondiabatic, in marked contrast with results obtained for Co(phen) 3 3+ as the oxidant. Linear kobsd VS. [Co(EDTA)--] plots are reported for the Co(EDTA)-- oxidation of cuprous stellacyanin at pH 10 (k = 8.9 M--1 sec--1; 25.0, pH 10, mu 0.5 M (carbonate); DELTaH not equal to 11.3 kcal/mol, deltaS not equal to = -16 cal/mol-deg) and at pH 7 in the presence of excess EDTA (k = 21.2 M--1 sec--1; 25.1 degree, pH 7.0, mu 0.5 M (phosphate), [EDTA] tot = 5 X 10(--4) M; deltaH not equal to = 5.9 kcal/mol, delta S not equal to = --33 cal/mol-deg). It is concluded that Co(EDTA)-- adopts an electron transfer mechanism similar to that preferred by Co(phen)33+ under conditions where the oxidant is prevented from binding strongly to reduced stellacyanin.


Plasma amino acid levels in Huntington's chorea.

Concentrations of plasma amino acids in nine patients with Huntington's chorea and nine control patients were studied while diet and drug therapy were controlled. Significantly low values for threonine, alanine, isoleucine,  leucine, lysine and histidine were found in the Huntington's chorea patients. However, since different investigators have failed to establish a consistent pattern of abnormality, it is considered that the findings are probably due to non-specific factors.


Evaluation of the thrombogenic potential of three types of arterial graft studied in an artificial circulation.

The thrombogenic potential of woven Dacron, expanded polytetrafluoroethylene (Gore-Tex) and gluteraldehyde-treated human femoral artery (HFA) grafts was evaluated using an artificial circulation. The differences in the platelet parameters (platelet count, adhesion and aggregation) and scanning electron microscopy (SEM) appearances before and after perfusion were used as indices of thrombogenicity. Platelet counts fell significantly with woven Dacron, moderately with HFA and no fall was observed with Gore-Tex. Platelet aggregation was unaffected by any graft, but platelet retention was significantly marked in woven dacron. SEM studies revealed minimal adherence of platelets to Gore-Tex and HFA endothelium, but marked adherences to woven Dacron, accompanied by morphological changes of the adhered platelets. In an artificial circulation, of the three types of arterial graft, woven Dacron has the greatest influence on platelet function, indicating significant thrombogenicity, whereas HFA is intermediate in its effect and Gore-Tex appears to have the lowest thrombogenic potential.


The use of peritoneoscopy in the detection of liver metastases.

Peritoneoscopy was carried out in 352 cancer patients with clinical suspicion of liver involvement in most cases. Principally because of patient discomfort, adequate liver biopsy was obtained in only 66% of 240 patients who underwent peritoneoscopy under local anesthesia while, under general anesthesia, biopsies could be taken in 90% of 112 patients. When the liver was macroscopically free of disease, the yield of positive peritoneoscopy was minimal regardless of the number of blind deep biopsies. Peritoneoscopy provided histologic demonstration of hepatic invasion in a total of 55 patients. Seven false-negative examinations out of 19 negative peritoneoscopies (36%) were identified by subsequent laparotomy or autopsy within 2 months. These preliminary data, although difficult to interpret in terms of accuracy of the method, point to the possible contributions of peritoneoscopy in detecting liver metastases.


Drug-induced changes in DNA fluorescence intensity detected by flow microfluorometry and their implications for analysis of DNA content distributions.

Chicken erythrocytes, which contain less DNA than mammalian diploid cells, were used as an internal standard to control instrumental and staining variables during flow microfluorometric analysis. With the DNA stain, mithramycin, and with an EPICS II flow microfluorometer, ratios between the modal G1 fluorescence of experimental cells and that of chicken erythrocytes were determined. The results indicate that unperturbed cell populations of L1210 and HeLa cells in vitro and L1210 ascites cells in vivo have relatively stable fluorescence ratios, although there is a significant difference between the ratios of one L1210 cell line in vitro and another in vivo. In contrast, L1210 ascites treated in vivo with different schedules of cyclophosphamide and Adriamycin showed wide fluctuations in the fluorescence intensity ratios for 96 hr after treatment. Also, differences in the fluorescence ratios were observed between less advanced and more advanced L1210 ascites after treatment with the same schedule. These effects indicate an alteration in DNA staining with mithramycin, brought about by drug treatment that could seriously affect the interpretation of DNA histogram data. Nevertheless, changes in mithramycin staining may prove to be a very important probe to detect persistent drug effects.


Biological and biochemical properties of the 2-hydroxyl metabolites of 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea.

The lethal and bone marrow toxicity and antitumor activity of the cis- and trans-2-hydroxylated metabolites of 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU) have been correlated with their relative in vitro alkylating and carbamoylating activities. Both the isomers have considerably greater alkylating activity and shorter chemical half-lives than the parent compound and on a molar basis have greater antitumor activity against i.p. L1210 leukemia. However, in terms of molar doses resulting in the death of 10% of normal mice, the cis- and trans-2 isomers were 2- and 3-fold more toxic than was CCNU in normal mice. In comparing the antitumor activity produced by a maximum nonlethal dose for each compound, we found that the trans isomer had activity identical to that of CCNU (413 and 410% increased life span compared to control), and the cis isomer had considerably less (152%). Like chlorozotocin, both isomers possess low carbamoylating activity and increased water solubility, two features that have been considered possible contributors to the bone marrow-sparing character of chlorozotocin. However, in the murine model the human bone marrow colony formation (CFU-C) assay neither hydroxylated metabolite of CCNU was associated with reduced myelotoxicity.


Activation of suppressor cells by syngeneic tumor transplants in mice.

Spleen cells from mice with syngeneic tumor transplants are hyporesponsive in mixed-lymphocyte culture. The hyporesponsiveness is due to the activation of suppressor cells. Spleen from tumor-bearing mice, treated with mitomycin and added to normal mixed lymphocyte culture (with responding cells syngeneic to the added cells), inhibits proliferation of the responding cells. Suppressor activity in the spleen cells can be detected as early as 5 days after s.c. transplantation of P-815 mastocytoma into DBA/2 mice. Tumor cells placed in cell-impermeable i.p. diffusion chambers can also activate splenic suppressor cells. Suppressor cells can be activated in syngeneic mice by (DBA/2) P-815 cells, by (C3H) L25-cells, or by recent (C57BL/6) methylcholanthrene-induced tumors. The latter tumors retain their ability to activate suppressor cells after passaging in syngeneic mice. Only one tumor, induced with methylcholanthrene in DBA/2 mice, failed to activate suppressor cells. Suppressor activity in the spleen cells from mice with 20-day s.c. tumor transplants is not reduced after removal of glass-adherent cells. Suppressor activity is significantly decreased after removal of thymus-derived cells with anti-theta treatment and complement.


Experimental right ventricular hypertrophy and failure in swine.

Following pulmonary arterial constriction, 22 pigs developed right ventricular failure and 10 right ventricular hypertrophy. Comparing results with those obtained in 21 normal pigs (no pulmonary artery constriction), it was found the right ventricular peak systolic and end-diastolic pressures, cardiac output, and right ventricular weight distinguished hypertrophied and failing hearts from normal ones. Cardiac output was lower in failing hearts than in hypertrophied hearts and was the only variable which significantly differentiated between these two groups. Left ventricular pressures and left ventricular and right ventricular dP/dtmax, and maximum [(dP/dt)/P] did not distinguish failing from hypertrophied hearts. Left ventricular pressure, maximum [(dP/dt)/P] and left ventricular and right ventricular dP/dtmax were not significantly different in the three groups.


Non-invasive electromagnetic measurement of the peripheral pulsatile blood flow: experimental study and clinical applications.

The non-invasive electromagnetic blood flowmeter described in this paper allows us to measure pulsatile flow through a limb. The limb is placed in a magnetic field and the blood flow rate induces electromagnetic forces which are detected at the skin surface with ECG electrodes (Faraday's law). A special computer technique is necessary to isolate the signal from artefacts (local ECG, BCG, EMG). In vitro calibration is performed using a circulatory model and in vivo using mongrel dogs. Its validity is assessed by comparing the results with the responses obtained from the invasive electromagnetic flowmeter. Sources of error in the measurement such as blood composition (Na+, K+), haematocrit (45% to 29%), and venous flow are reported here. The results indicate that the method is reliable, easy to utilise and offers a unique non-invasive way of measuring true pulsatile blood flow rate in humans. Various clinical applications are discussed for possible use of the device.


Prediction of multivessel disease after inferior myocardial infarction.

We correlated clinical parameters with angiographic findings in 108 men with a previous isolated inferior myocardial infarction, to determine if these parameters could predict accurately which patients had multivessel disease. Of 71 men in angina class 2-3, 42 had three vessel disease versus only seven of the 37 who were either asymptomatic or angina class 1 (P less than 0.001). Multivessel disease was present in 35 of the 36 who had anterior ST-T abnormalities at rest (P less than 0.001) and 16 of the 17 with cardiomegaly. Among men 55 years and older, the incidence of multivessel disease was 94% compared to 70% in men less than 55 (P less than 0.03). We conclude that functional angina class, age, and the presence of resting anterior ST and T abnormalities are highly predictive of associated left system disease in survivors of inferior infarction.


Echocardiographic diagnosis of left ventricular hypertrophy.

Echocardiograms were obtained on 27 adults with electrocardiographic criteria of left ventricular hypertrophy (LVH) to determine how echocardiograms might best identify LVH. Both the left ventricular (LV) posterior wall thickness and interventricular septal thickness were found by echocardiography to be increased (greater than or equal to 12 mm) in only 13 of 27 patients (48%) with LVH. The LV was dilated (greater than or equal to 58 mm) in the absence of posterior wall thickening in 9 of 27 patients (33%). The LV mass, estimated from standardly measured dimensions, was increased (greater than 200 g) in 21 of 27 patients (78%) and when measurements were made by the Penn method, mas was increased in all patients. These observations indicate that the echocardiographic estimation of LV mass is a more sensitive indicator of LVH than LV posterior wall and septal thickness. Since LVH is defined as an increased mass of LV muscle, these observations are consistent with this fundamental definition of left ventricular hypertrophy.


Endocrine studies in male pseudohermaphroditism in childhood and adolescence.

The clinical and endocrine features of fifty cases of male pseudohermaphroditism and XY karyotype aged from 6 months to 20 years have been studied. Thirty-two subjects were pre-pubertal and eighteen, of whom ten developed gynaecomastia, were pubertal. A definite aetiology was established in 12%. 4% had deficient testosterone biosynthesis and 8% mixed gonadal dysgenesis. In the remaining 88% an aetiology of androgen unresponsiveness at the target areas is suggested. Hormonal investigations in these subjects showed that before puberty basal plasma testosterone and oestradiol were slightly but significantly elevated, whereas testosterone response to stimulation with human chorionic gonadotrophin was significantly diminished. In the pubertal subjects basal plasma testosterone, oestradiol, serum binding capacity of testosterone-oestradiol binding globulin and basal plasma LH were significantly elevated. These hormonal features in pubertal or post-pubertal male pseudohermaphrodites appear to be characteristic of androgen unresponsiveness. The presence in addition of elevated basal plasma testosterone and oestradiol in the pre-pubertal subjects suggest that some of these findings may be presented from early childhood.


Sanfilippo A syndrome in the fetus.

A family is reported in which Sanfilippo A syndrome affected three siblings: the proband and twin premature infants. The feasibility of intrauterine diagnosis of mucopolysaccharidoses (MPS) Type IIIA, was demonstrated by the excessive accumulation of 35SO4-mucopolysaccharides in fibroblasts cultured from amniotic fluid obtained by amniocentesis Cross-correction studies and enzymatic analysis of cultured skin fibroblasts from the proband and the infants revealed the absence of the MPS IIIA correction factor, heparan sulfate sulfatase. However, when the premature infants expired shortly after birth, no central nervous system histopathology or ultrastructural abnormalities were found. From these observations it would appear the the third trimester fetus with MPS type IIIA has little CNS involvement.


Evidence of both anti-GBM and immune complex mediated pathogenesis in the initial phase of Goodpasture's syndrome.

A 19 year old women presented with pulmonary hemorrhage, rapidly progressive glomerulonephritis and antiglomerular basement membrane antibody in her serum. In the initial phase of the disease a double layered linear deposit of IgG was seen on the glomerular capillary basement membrane together with a finely granular or focal and lumpy single layer deposit of C3 and numerous small subepithelial electron dense deposits. These findings are suggestive of the simultaneous occurrence of anti-GBM and immune complex glomerulonephritis.


Structural determinants of disease and their contribution to clinical and scientific progress.

The recognition of structural correlates with disease provided a release from the Hippocratic humors which dominated the approach to medicine for more than a thousand years. During recent decades an increasingly reductionist approach to structure has enabled such correlations to extent from those easily visible to the naked eye to those who resolution requires sophisticated electron microscopy. Although recognition of clinical--pathological correlations has evident advantages, there are instances, such as diabetes mellitus, where such recognition appeared to lull investigators into believing that the pathogenic mechanisms underlying the disease had been found. Moreover, the recognition of a structural abnormality, such as the extra chromosome in Down syndrome, may sometimes have delayed rather than accelerated research on metabolic aspects of the disease. The advances in medical understanding that have proceeded from a correlation of structure with function have enabled diseases to be investigated at different levels. This has led to a more sophisticated redefinition of the meaning of clinical--pathological correlations and to the surprising finding that immense structural variability can be tolerated by the human organism without its function being recognizably perturbed. Indeed, some molecular variations of structure lead to a distinct biological advantage under singular environmental conditions. Finally, where molecular structure is known in its most intimate detail, the distinction between structure and function becomes less easy to discern.


Human parturition.

The roles of fetal adrenal hormones, of maternal oestrogen and progesterone concentrations, and of uterine prostaglandin synthesis are considered in relation to the onset of human labour. Evidence that the fetal adrenal may be involved in the onset of labour is discussed with particular reference to both cortisol and dehydroepiandrosterone sulphate. Towards the end of pregnancy circulating concentrations of both unconjugated and conjugated oestrogens increase, and evidence for the presence of arylsulphatase (EC 3.1.6.1) in various intrauterine tissues suggests that the conjugates are potentially active. The rise in oestrogens and a concomitant drop in progesterone during the last few weeks of pregnancy may play a facilitatory role in the onset of labour. The changes in steroid concentrations are considered in relation to the promotion of prostaglandin synthesis leading to labour. Concentrations of prostaglandins in amniotic fluid show only minor changes before the onset of labour compared to those found with the onset of and during labour. Evidence for a local control of prostaglandin synthesis within the uterus is presented.


[Serum IgE in skin diseases of children (author's transl)].

Using the paper-radioimmunosorbens test (PRIST) normal values were obtained from 200 non-allergic children. The IgE level was aged-dependent, in part with significant differences between various age groups. These normal values were compared with those in children with seborrhoeic dermitis, constitutional neurodermatitis, parasitoses, urticaria, Quincke oedema, Schönlein-Henoch purpura, pityriasis rosea, multiform exudative erythema, erythema nodosum and infantile papular acrodermatitis. Half-life of IgE in neonates was 16.2 hours.


Luteinized unruptured follicle syndrome: a subtle cause of infertility.

A clinical description of luteinized unruptured follicles is presented. This abnormality in ovulation is characterized by normal endocrinologic presumptive signs of ovulation: biphasic basal body temperature curves, secretory endometrium, and laboratory evidence of progesterone production by elevated urinary pregnanediol or plasma progesterone levels. In a group of 102 such infertile women, laparoscopy performed 3 to 5 days after apparent ovulation revealed the absence of a corpus hemorrhagicum in 30 women, and the absence of a sigma on a corpus hemorrhagicum in an additional 32 women. These findings were evidence that a follicle had not ruptured and an ovum had not escaped. Of 28 patients undergoing follicular stimulation with clomiphene citrate or human menopausal gonadotropin after this diagnosis, 15 conceived.


Progressive symmetric erythrokeratodermia.

Progressive symmetric erythrokeratodermia is a rare disorder of keratinization manifested by hyperkeratotic plaques on an erythematous base. Familial cases have not been reported. The following is, to our knowledge, the first report of a case from America, as well as the first involving a black child.


Induction of abortion by intrauterine administration of prostaglandin via laparoscopy with concurrent sterilization.

This is a preliminary report of a method of inducing abortion at 12 to 15 weeks' gestation, a period when none ot the currently available methods are wholly satisfactory. The authors investigated both prostaglandin F2ALpha (PGF2alpha) and 15-methyl PGF2alpha injected intramyometrially and intraamniotically under laparoscopic visualization prior to sterlization as a means of inducing abortion. They conclude that the results from this small series (13 patients) indicate that this is a practicable method for abortion combined with sterilization for patients at 10 to 15 weeks' gestation and recommend that further comparative studies be undertaken.


Laparoscopic sterilization after "spontaneous" abortion.

One hundred eighty-nine patients were evaluated to assess the safety of laparoscopic sterilization after completion of a "spontaneous" abortion by curettage. Some of the "spontaneous" abortions had probably been illegally initiated outside the hospital and were septic. The laparoscopic sterilization procedures used tubal rings to achieve tubal occlusion. They were performed 4 to 48 hours after completion of the abortion. Patients were discharged from the hospital on the day the procedure was performed. Surgical difficulties and complications associated with the laparoscopy were infrequent. Undergoing a curettage prior to sterilization with tubal rings by laparoscopy did not appear to result in increased rates of complications.


[A preliminary study on parietal stress decay in patients with left ventricular hypertrophy (author's transl)].

In order to determine equatorial and radial stresses at various depths of left ventricular wall, left ventricular pressures and echocardiographic dimensions were elaborated using the formula proposed by Timoshenko. Patients with concentric left ventricular hypertrophy (LVH) have minor end-diastolic and end-systolic decay of radial and equatorial stresses, compared to patients with eccentric LVH. The total stress affecting left ventricular wall is significantly minor in telediastole than in telesystole.


[Masked depressions in obstetrics and gynaecology (author's transl)].

The development of psychosomatic symptoms can not exclusively by explained by fear and rejection of fear. We showed in another paper that other emotions but fear can lead to functionae symptoms in obstetrics and gynaecology. The recognition of depressions and masked depressions as the basis for many psychosomatic symptoms in gynaecology can open therapeutic possibilities for many cases which were refractory to treatment. The prescription of antidepressant medication often results in rapid relief of the symptoms. Psychotherapy for the discovery and for restructuring becomes possible in many cases after a successful course of anti-depressant medication.


Genetic and physiological characteristics of a slow-growing circadian clock mutant of Neurospora crassa.

A circadian clock mutant of Neurospora crassa with a period length of about 25.8 hours (4 hr longer than wild type) has been isolated after mutagenesis of the band strain. This mutant, called frq-5, segregates as a single nuclear gene, maps near the centromere on linkage group III, and is unlinked to four previously described clock mutants clustered on linkage group VII R (Feldman and Hoyle 1973, 1976). frq-5 differs from the other clock mutants in at least two other respects: (1) it is recessive in heterokaryons, and (2) it grows at about 60% the rate of the parent band strain on both minimal and complete media. Double mutants between frq-5 and each of the other clock mutants show additivity of period length--two long period mutants produce a double mutant whose period length is longer than either of the two single mutants, while a long and a short period double mutant has an intermediate period length. Although slow growth and long periodicity of frq-5 have segregated together among more than 300 progeny, slow growth per se is not responsible for the long period, since all the double mutants have the slow growth characteristic of frq-5, but have period lengths both shorter and longer than wild type.


[Dosage effect of the cytoplasmic superoxide dismutase (SOD-1) gene in the erythrocytes of Down's syndrome patients].

The activity of cytoplasmic superoxydase (SOD-1) was studied in erthrocytes of 17 patients affected with Down's syndrome (trisomy 21) and in 26 healthy persons. A 1.56-fold increase of the enzyme activity was observed in the group of patients as compared with the control group. This could be explained as the dosage effect of the corresponding gene located in the chromosome 21.


[Geno- and phenotypic distribution by the ABO and Rhesus systems in the population of Minsk].

The distribution of blood groups of the ABO system is different in the rhesus-positive and rhesus-negative subpopulations. An increasing frequency of the phenotype A2 in the rhesus-negative subpopulation is observed. The calculation of the gene frequency reveals a deficiency of genes A1 and O, and the increasing frequency of genes A2 and B in the rhesus-negative part of the population. On the whole for the inhabitants of Minsk the ratio of the phenotypes A2/A1 differs significantly from the corresponding index for the republic as a whole, which is the evidence of a significant migration of the population in the capital of Byelorussia. The observed correlative dependences in the distribution of the genes of ABO system and of the rhesus system permit to recommend the accomplishment of a scientific search between the hereditary pathology and the susceptibility to different diseases and blood groups taking into consideration the rhesus appurtenance of the populations studied.


Degradation of human fibrinogen by plasmin: isolation and partial characterization of an early degradation product.

The incubation of human fibrinogen with plasmin gives rise to an early degradation product with a molecular weight of 63,000, calculated by polyacrylamide-gel electrophoresis sodium dodecyl sulfate (PAGE-SDS); this product is resistant to the action of the plasmin for short incubation times and represents 1 +/- 0.2% of the original quantity of fibrinogen. The fragment was isolated from the incubated mixture by gel filtration and has a no-identity reaction with fibrinogen fragment D against fibrinogen fragments D antiserum. The reduction gives rise to three polypeptidic chains with molecular weights of 36,000, 17,000 and 10,000 (calculated by PAGE-SDS). Study of the carbohydrate content of these polypeptidic chains from the reduced 63,000 MW fragment indicates that there is only one electrophoretic region which contains periodic acid-Schiff (PAS) positive material. As the 63,000 MW fragment has a reaction against fibrinogen fragment D antiserum and also contains PAS positive material, according to Pepper and co-workers, it can be taken to arise from the HOOC-terminal region of fibrinogen.


Experimental defibrination and bothropase: a study on the fibrinolytic mechanism in vivo.

Fibrinogen was depleted in dogs injected with a single dose of bothropase even if pretreatment followed by a continuous infusion of antifibrinolytic drugs was performed during defibrination. The activation of the fibrinolytic system as a secondary effect of the defibrination syndrome induced by bothropase injection was blocked completely by aprotinin (Trasylol) but not by EACA. Plasmin activity in spite of the inhibition of plasminogen activator suggests that, either an excess of activator is released in circulation or a plasmin-antiplasmin complex is dissociated by the circulating fibrin, according to the hypothesis of Ambrus and Markus [1], and Back et al. [4] for the mechanism of fibrinolysis in vivo. An experimental model is suggested for the study of the fibrinolytic mechanism in vivo, by the association of defibrinating agents, antivenom and antifibrinolytic drugs.


[Fibrous dysplasia of the skull--a radiological, computer tomographic and scintigraphic study (author's transl)].

The diagnosis of fibrous dysplasia (Jaffé-Lichtenstein) of the skull can generally be made by plain films, occasionally supplemented by tomography. The differential diagnosis from sclerosing meningiomas in the anterior fossa can usually be made by cranial computer tomography. Cerebral scintigraphy does not play an important part in the differentiation from a meningioma. Total body scintigraphy, on the other hand, is useful, particularly in distinguishing between monostotic and polyostotic bone conditions. Invasive neurological methods, such as cerebral angiography, are in our opinion unnecessary unless there are definite clinical focal cerebral symptoms in the presence of a normal computer tomogram.


A new finding in bone scintigram 3 cases of defect findings.

Three cases in which accumulation in the lesion was less than that in the normal area by bone scintigraphy using 99m TC pyrophosphate or methylenediphosphate were reported.


Anatomy and radiology of a bone island.

Anatomoradiological study, on an amputation specimen for osteosarcoma from a 13 year old girl, of a bone island in the distal femoral epiphysis. It consists of a mature bone core caused by localized remodeling of the spongeous trabeculae, old but still slightly active, clearly different from a bone infarct, a tumoral or infectious process. Discussed the interest of a precise radiological study for making the diagnosis (particularly xerography), the risks of error involved in scintigraphy and the utility of a block excision if a biopsy is performed.


[Band-like osteoporosis (author's transl)].

Atrophy of the skeleton due to inactivity may have two types of radiological change: diffuse decalcification and band-like porosis in the zones involved by previous growth. The type of decalcification is evidently determined by the vascular anatomy of the bones. This results in localised differences which are independent of age, the type of trauma or the duration of immobilisation.


[Intradiploic leptomeningeal bone cysts (author's transl)].

A temporo-occipital lepromeningeal cyst was observed in a 70-year old man; it presented as a translucency with serpigenous outline and a sclerotic margin. There was intra- and extra-cranial bulging of the calvarium. The extremely long latent period was unusual, since the original injury had occurred at the age of four years and the probable first clinical symptoms (giddiness) had manifested themselves at the age of 65 years. Since the correct diagnosis had been made at the age of 70, there was a 66 year interval. Vertebral angiography confirmed the intracranial expansion. From the history, clinical findings and extensive radiological investigations, an epidermoid or an intradiploic leptomeningeal cyst was considered in the differential diagnosis. At surgery the latter was confirmed.


A variation of scimitar syndrome.

Variations of the scimitar syndrome have been reported previously, the most common being the absence of systemic arterial supply to the right lung and varying degrees of lung hypoplasia. Anomalous scimitar-like veins draining the entire right lung and connecting normally to the left atrium have been reported. The case presented is unique in that there is marked hypoplasia of the right bronchial tree and a large bronchogenic pulmonary cyst without dextroposition of the heart and anomalous systemic arterial supply to the right lung.


[The investigation of the renal veins in the nephrotic syndrome in adults (author's transl)].

The problems in the radiological investigation of adults with the nephrotic syndrome are illustrated by three patients. In one it was due to spontaneous thrombosis of the cava, in the second there was renal vein thrombosis from self-administered overdose and the third patient had glomerulonephritis without renal vein thrombosis. In view of the therapeutic implications, radiological investigation of the renal veins is essential and should be carried out in every patient with the nephrotic syndrome.


[Morphological changes following arterial catheter embolisation of the kidney (author's transl)].

Arterial catheter embolisation of the kidney is used as a form of treatment for uncontrollable haematuria due to inoperable tumours of the kidney or bladder; it can also be used as a preoperative measure for operable renal tumours in order to make surgery easier. Time-related morphological examinations are not yet available in the literature. The histological changes in the tumour-containing kidney are described in four patients at two, six, twelve and thirteen days after Histo-acryl embolisation. After embolisation there are wedge-shaped anaemic infarcts and extensive necrosis in tumour and normal kidney with leukocytic demarkation and reabsorption of cellular inflammatory material. Complete necrosis of the tumour was not found. There is therefore not complete destruction of the tumour as a result of arterial catheter embolisation, even when the veins are thrombosed by tumour.


[Concentration of antibiotics in long term embolized kidneys (author's transl)].

In 7 dogs renal arterial embolization was performed using Seldinger technique and inflatable Swan Ganz balloon catheter by injection of methyl methacrylate/butyl 2 cyano acrylat. Six months later tissue concentration of cefazolin was measured in the embolized and kontralateral kidneys and in serum two hours after a short infusion of 1 g of Cefazolin. The concentrations in serum and homogenized kidney tissue were determined by means of the agar well diffusion method. Angiography studies showed no vessels in the embolized kidneys. Histologically there was total atrophy of the tissue especially the tubular system. The mean tissue level of Cefazolin in the embolized kidney was 70% of serum. The tissue concentration was always above 10 microgram/g. This concentration is high enough to inhibit nearly all strains of E. coli, Klebsiella, Salmonella, Shigella and a large part of Proteus mirabilis, whereas nearly all gram-positive bacteria are inhibit at a lower concentration. Value and necessity of antibiotic therapy in embolized kidney cancer is discussed.


[The jet phenomenon in the bladder (author's transl)].

The jet phenomenon seen during entry of urine into the bladder is illustrated by the urograms of a number of patients. Its diagnostic significance is discussed. Experimental models and theoretical considerations have shown that the jet is determined by the kinetic impulse; for a 1:10 mixture of contrastmedium and water, the jet is visible at all jet diameters and jet speeds at the orifice provided flow at the centre of the jet has fallen to about 0.073 m/s. Transferring these findings to man, and bearing in mind the functional and anatomical peculiarities and pressure differences between the lower ureter and the bladder, it becomes apparent that this phenomen can be produced under physiological conditions.


[Phaeochromocytomas in neurofibromatosis (author's transl)].

Neurofibromas and phaeochromocytomas derive ultimately from ectodermal tissue. It is therefore not surprising that there is an increase in the incidence of phaeochromocytomas in Recklinghausen's neurofibromatosis. When investigating hypertension in patients with neurofibromatosis one must therefore consider the possibility of a phaeochromocytoma as well as vascular involvement by neurofibromatosis. Two patients with neurofibromatosis and phaeochromocytomas are described.


[Exogenous allergic alveolitis during childhood (author's transl)].

Four children aged 31/2 to 13 years, who developed allergic alveolitis as a result of antigenic exposure in a domestic situation are described. The clinical symptoms consisted of gradual onset of increasing dyspnoea, chronic cough and weight loss. Lung function tests showed characteristic reduction in vital and diffusion capacity. Serologically, antibodies against animal antigens were demonstrated in all patients. Radiologically there were pronounced reticular changes in both lungs affecting mostly the middle and lower zones. Occasionally there were miliary and sometimes confluent shadows. In differential diagnosis, virus infections, pneumocystis carinii, fungus infections, dermatomyositis, sarcoidosis and tuberculosis had to be considered. Apart from a short course of steroid treatment, it is necessary to make certain that the children are no longer exposed to the causal antigen. Under these conditions the prognosis is good, but otherwise the disease, as in adults who are continuously exposed to the antigen, results in pulmonary fibrosis.


[Disseminated branching pulmonary ossification combined with heterotopic bone formation in the muscles (author's transl)].

Idiopathic disseminated pulmonary ossification is a rarely recognised condition in which small pieces of marrow containing bone are found in the interstitial tissues of the lungs. The case reported here is unique in showing combination with myositis ossificans circumscripta. The extent of the lung changes was also unusual. The clinical, radiological and histological findings are compared.


[Contrast extravasation from the pelvicalycine system during infusion pyelography (author's transl)].

Rupture of the pelvicalycine systems is occasionally observed during infusion pyelograms. Twenty such cases are reported. The usual cause was the presence of stones in the ureter leading to urinary tract obstruction and acute pressure rise in the pelvicalycine system. The anatomical and physiological basis for the rupture is outlined. Subsequently our own cases are described. In 13 patients rupture occurred without previous renal abnormality; in these there was always spontaneous healing once the obstruction had been removed. In five patients rupture occurred in a previously damaged kidney; the changes at the point of rupture and the subsequent possible complications are described. Two cases should be mentioned in detail; in these, rupture persisted and lead to the development of a retroperitoneal pseudocyst. Only one similar case is reported in the literature since 1966.


[An evaluation of the risks of pulmonary scintigraphy (author's transl)].

Scintipulmography is a method for testing cardiopulmonary function in diagnosis and control. Standardized X-ray examinations are preferable from a radio-hygienic view. In perfusion scintigraphy microspheres are better for examining terminal vessels and are of lower risk. Short-lived radionuclides, e.g., 99mTc or 113mIn, should be used as markers. Recent physiologic findings indicate injection by bolus during the middle phase of inspiration while the patient holds his breath. General and individual risks and the diagnostic advantages of this method are discussed. Normally, the advantages outweigh the risks, as demonstrated in a case of artificial pulmonary embolism. For high-risk patients, however, inhalation scintigraphy should be made available as a standard radiologic method.


[A new method for direct measurement of the aortofemoral pressuregradient in connection with translumbar aortography (author's transl)].

A new method to measure simultaneously and directly the aorto-femoral pressure-gradient is described. The study can be performed in connection with translumbar aortography. In addition of being easy to perform the following advantages can be obtained by using this method: 1. Additional angiographic projections are unnecessary. 2. Informations regarding the absolute value of the aortofemoro pressure-gradient at rest and after maximal vasodilatation is obtained.


[Intravenous arteriography with xerography: a non-invasive method for the demonstration of peripheral vessels (author's transl)].

Acceptable arteriograms of the extremities can be obtained by rapid manual injection of contrast into a cubital vein, using a xeroradiographic technique. The indications and value of the method have been demonstrated in 62 examinations. The new technique constitutes a non-invasive arteriographic method suitable for demonstrating limited vascular lesions in the extremities.


[Arteriography in old age--an assessment of the risks and technique (author's transl)].

Of 1,263 arteriograms performed over a period of two and a half years, there were 166 patients aged 70 years or more. The indications, angiographic findings and results have been examined retrospectively. In seven patients only was it impossible to carry out the examination because of advanced vascular disease in the pelvis. Most examinations could be performed as planned. In three patients there were minor complications without consequence. In one patient the external iliac artery was occluded following catheter angiography, but this was disobliterated during a subsequent operation which would have been carried out in case. The severity of vascular calcification as seen on plain films gives no indication regarding the difficulties during catheterisation. If aortography by catheter is impossible, we demonstrate the aorta retrogradely by a counter-current technique, using the Hettler instrument. The Hettler technique does not increase the risk and has certain advantages for the examination of older patients.


[The post-traumatic arteriovenous symptom complex (author's transl)].

The post-traumatic arteriovenous symptom complex is an atypical manifestation of saccular arterial and arteriovenous aneurysms. Prominent in this condition is venous obstruction accompanied by abnormal arterial flow. Treatment for this condition is unsatisfactory, and the complex consists of a trio of venous and arterial obstruction and resistance to therapy. The complex can be recognised by a typical pulse form and the diagnosis can be confirmed by angiography of the affected limb.


[Radiological studies of the anatomy of the vena semicircularis and the Giacomini vein (author's transl)].

The course and variations of the vena semicircularis and Giacomini veins has been studied. It appears that, despite the large variations encountered, three major types can be identified.


[The use of metrizamide for peripheral angiography. A comparative investigation (author's transl)].

In peripheral arteriography in 20 patients the non-ionic contrast medium metrizamide (amipaque) and the ionic megluminamidotrizoat (angiografin) was injection. Following amipaque no pain reactions were observed but angiografin caused severe pain in 11 patients and light pain in 3 patients. The diagnostic information of the two contrast media was the same.


[Thoracic myelography with water-soluble contrast medium (metrizamide) (author's transl)].

The use of water-soluble contrast media for the thoracic myelography has become possible with the introduction of Metrizamide (Amipaque). A modification of the technique used with oily contrast media results in excellent demonstration of detail of intraspinal structures and of pathological changes. The medium is well tolerated and does not cause irritation of the spinal cord; the results must be regarded as excellent.


[Selective spinal arteriography in vertebral and paravertebral disease processes (author's transl)].

Selective arteriography was performed on 15 patients with vertebral and paravertebral diseases. This procedure is particularly justified if the bone changes are of possible vascular origin, if the presence of arterial involvement by neoplasm is to be determined, or if one wishes to judge the extent of intraspinal or paravertebral spread of a spinal tumour. Up to a point, angiography may indicate the histology and malignancy of a mass. In the diagnosis of vertebral and paravertebral disease, selective spinal arteriography would come into competition with computed tomography. Either procedure should only be used if conventional methods are incapable of solving the diagnostic problem.


[Standard report forms in radiological diagnosis (author's transl)].

Standard report forms have been designed for excretion urography, gastro-intestinal examinations, cholecystography, myelography, trauma and surgical emergencies; the findings, which are printed on the form, are marked and the conclusion is then written by hand. If used for a limited number of examinations with recurring problems, these report forms results in a considerable reduction in effort and simplifies routine diagnosis.


Rare-earth and calcium tungstate intensifying screens, a comparative study of relative speed, radiation doses and resolving power.

Using calcium tungstate (CaWO4) and rare-earth intensitying screens, their relative speeds were compared in exposures of a pelvis phantom, and radiation doses were measured at several kilovoltages. Using a slit pattern, the resolving power was assessed by a modified contrast-frequency response and by direct viewing of the radiographs. In the comparison of the rare-earth and CaWO4 screens the following differences were found: (1) A great reduction in the radiation dose can be achieved by the use of the fast screen types, yet at the cost of some loss of resolving power; (2) the film contrast can be improved by a reduction in the kilovoltage without an increase in the radiation dose; (3) the short exposure times and the characteristic course of the dose curve necessitate an adjustment of the phototimer or a change of the control unit; and (4) with the slower screen type tested, the resolving power can be improved without increasing the radiation dose.


[Diaphragmatic metastases in ovarian neoplasms].

Laparoscopic examination is of considerable importance for the prognosis of ovarian carcinoma, especially the detection of metastases in the diaphragm and other abdominal areas. 65 patients with ovarian carcinoma underwent laparoscopy; in 31 cases metastases in various abdominal areas were found, in 25 cases also the diaphragm was involved. This high incidence seems to be remarkable.


[Tryptophan metabolism and oligophrenia (author's transl)].

After dealing with the biochemistry of tryptophan metabolism the most important results obtained in humans are presented. Special emphasis is given to the hereditary defects of tryptophan metabolism associated with mental retardation and convulsions due to lack of pyridoxine. The author's findings demonstrate the existence of a hereditary disturbance of the tryptophan metabolism via kynurenine in a certain part of oligophrenic patients. This metabolic defect can be controlled by high doses of vitamin B6. Furthermore investigations conducted with a view to interpreting these results are discussed, especially the determination of kynureninase activity, serotonin blood levels and pyridine nucleotide synthesis.


Compliance with 504 regulations need not require costly renovation.

By June 3, 1980, all hospitals receiving federal funds must have completed plans to make facilities accessible to the handicapped. In many instances, doing so will require hospitals to renovate existing facilities. However, through careful interpretation of the law and creative planning, hospitals can avoid making expensive, unnecessary building modifications.


Rehab Act calls for new approach to interviewing job applicants.

Compliance with regulations implementing section 504 of the Rehabilitation Act will require hospitals to revise hiring practices. Personnel administrators will need to examine employment screening and selection devices, especially, to ensure that they do not discriminate against handicapped applicants.


Dietary enhancement of CNS neurotransmitters.

Feeding choline, a precursor of the neurotransmitter acetylcholine, has been shown experimentally to increase brain acetylcholine release and enhance cholinergic function A subsequent trial of oral choline ameliorated symptoms in patients with tardive dyskinesia, which has been linked to cholinergic dysfunction. The findings suggest a role for dietary precursors in treating diseases associated with neurotransmitter abnormality.


Kinetics of 21-trisomic lymphocytes. I. In vitro response of 21-trisomic lymphocytes to PHA.

Data presented here on the quantitative 3H-thymidine incorporation into DNA, after PHA mitogenic stimulation, show that 21-trisomic lymphocytes are low-responders to PHA compared with the normal-diploid ones. Their responsiveness seems to decrease with the donor's age. Auto-radiographic studies clearly demonstrate that the fraction of labeled cells at the 72nd h of incubation is significantly smaller in the 21-trisomic lymphocyte population. The comparison of labeling indexes at different times of incubation (24, 48, 72 h) also indicate, in the same population, a slower increment of the portion of DNA-synthesizing cells. Discussing these data in the light of other's observations and recent progress in the knowledge of factors and mechanisms involved in the lymphocyte response to lectin mitogenic stimulus, it is suggested that differential distribution of T- and B- and/or T-cell subpopulations and a retarded cell induction time to proliferate may be two important factors negatively influencing the responsiveness of 21-trisomic lymphocyte population.


Evidence for the inheritance of silver-stained nucleolus organizer regions.

The inheritance of nucleolus organizer regions (NORs) was investigated by examining the degree of silver-staining in individual acrocentric chromosomes in two successive generations. The study was undertaken in six Down's syndrome children and their respective parents. Quinacrine fluorescent polymorphisms were used to identify individual acrocentrics and to determine which of the child's acrocentrics were informative as to parental homologue of origin. Of the 66 acrocentrics in the six children, 31 were informative. The correlation between the degree of silver=staining in the child's chromosomes and the respective parental chromosomes of origin was highly significant (P less than 0.001), with a correlation coefficient of 0.90. The results suggest that the degree of Ag-AS staining is characteristic for a particular chromosome and that this characteristic is an inherited property.


Regulation of lymphocyte proliferation by soluble products released by stimulated human lymphocytes.

Human lymphocytes pulsed with phytothaemagglutinin (PHA) release mitogenic factors and other lymphokines upon subsequent incubation in fresh medium. Such active supernatants (SUPs) were assayed for their capacity to modify the DNA synthetic response of human lymphocytes exposed to various mitogenic stimuli. Both non-fractionated peripheral lymphocytes and preparations of fractionated T cells were stimulated by active SUPs to DNA synthesis. In contrast, active SUPs reduced the responses of lymphocytes to allogeneic cells, PPD tuberculin and PHA. These reductions were most evident at the end of the culture periods. The results suggest that soluble products released by stimulated lymphocytes activate suppressor lymphocytes which inhibit proliferative responses of lymphocytes.


Behaviour of sheep-immunoglobulin-bearing and non-immunoglobulin-bearing lymphocytes isolated by nylon wool columns.

The technique of separating lymphocytes on nylon wool columns has been applied to sheep lymphocytes obtained from efferent lymph. The non-immunoglobulin-bearing (sIg-) lymphocytes which pass through the column were MLR-reactive, responsive only to T cell mitogens, and migrated in vivo like T lymphocytes described in rodents. Immunoglobulin-bearing (sIg+) lymphocytes were MLR-non-reactive, responsive to lipopolysaccharide and migrated in vivo like B lymphocytes in rodents. It is considered that the majority of sIg- lymphocytes obtained in this way are T lymphocytes and the majority of sIg+ lymphocytes are B lymphocytes.


Urethral tube graft in dogs: prosthesis of dacron lined silicone.

Silicone urethral prostheses lined with dacron velour were used for urethral replacement in eight mongrel dogs. In five of these prostheses the silicone was lined on the outside, and in three on the inside and outside. Seven dogs survived, one of which is still under observation at 10 months. Findings of excretory urography at the time of sacrifice were normal. In four cases urethrography showed significant flow around as well as through the prosthesis. In all instances study of urethral histologic sections showed complete encasement of the prosthesis by regenerated epithelium. A dacron lined silicone prosthesis can thus serve as an effective total urethral replacement in animals and may be used clinically in the future.


[Occupational dermatitis due to a yellow quinophthalone dye (solvent yellow 33: C.I. 47 000)].

An occupational allergic contact dermatitis caused by a yellow quinophthalone dye which is used for colored smokes in detonators was the reason for further studies of this case of allergy. Solvent Yellow 33 (German: Rauchgelb) belongs to the groups of quinaldine dyes which are produced by condensing quinaldine and phthalic anhydride. The dye itself is a mixture of 67% unsubstituted quinophthalone and 33% 6-methylquinophthalone (Colour Index 47 000). Quinophthalone derivatives are widespread in technology and industry and are used as yellow dyes in food (pudding), plastics (PVC), textile goods (nylon) and cosmetics (lipsticks, make-up). Cases of contact sensitization due to quinophthalone dyes are seldom recognized. Sensitizing experiments in guinea pigs were unsuccessful. Therefore Solvent Yellow 33 must be considered a weak sensitizer.


Histochemical definition of muscle fibre types in the trunk musculature of a teleost fish (cod, Gadus morhua, L.).

Cryostat sections incubated for myofibrillar ATPase, SDH, LDH, and alpha-GPDH as well as p-phenylene-diamine stained semithin sections were used to define muscle fibre types in the trunk musculature of the cod (Gadus morhua, L.). Three zones (superficial, intermediate, deep) containing different muscle fibre types are present within both epaxial and hypaxial parts of each myomere subjacent to the lateral line. Atypical relations concerning myofibrillar ATPase activity probably reflects instability of myosin during storage of frozen tissue. The histochemical reaction does not distinguish between myofibrillar and mitochondrial ATPase in cod muscle. Based on ATPase and SDH activities, seven different histochemical profiles of muscle fibres can be identified in trunk musculature of this teleost fish. Attempts to homologize these fibre types with those in cyclostomes or those in higher animals proved futile. The higher number of histochemically defined muscle fibre types in cod might be explained by developmental processes and an admixture of immature fibres throughout life.


"Red" fibres of pigeon pectoralis major muscle are "type II red".

On the basis of the histochemical activity of succinic dehydrogenase, only two fibre-types are distinguished in pigeon pectoralis major muscle. These are narrow "Red" and broad "White". The histochemical activity of myofibrillar ATPase was studied in these two distinct fibre-types. Both fibre-types showed high activity for the ATPase. "Red" fibres of pigeon pectoralis were not alkali-labile, at incubation pH 9.4, as were the "Type I" fibres of both avian and mammalian muscles. Again unlike "Type I" fibres, the "Red" fibres of pigeon pectoralis lacked the characteristic activation of acid-preincubated ATPase reaction. Pigeon pectoralis "Red" fibres are known to possess some characteristics of fast-twitch fibres (e.g. high fat, considerable phosphorylase, fibrillenstruktur myofibrillar arrangement, focal "en plaque" pattern of nerve endings). It is emphasized, therefore, that the pigeon pectoralis "Red" fibres are not equivalent to "Type I or slow-twitch", muscle fibres, but they are possibly "fast-twitch fatigue resistent or Type II Red" muscle fibres.


Cyanide- and hydroxamate-resistant respiration in Neurospora crassa.

Strain inl-89601 of Neurospora crassa respires exclusively by means of the mitochondrial cytochrome chain. The respiration of this strain is entirely inhibited by cyanide or antimycin A, the classical inhibitors of cytochrome chain respiration. When this strain was grown in the presence of chloramphenicol, however, two additional terminal oxidases were detected. One of these oxidases is inhibited by substituted hydroxamic acids and has been described previously. The second oxidase was not inhibited by cyanide or hydroxamic acid but was inhibited by azide in the presence of both cyanide and hydroxamic acid. This azide-sensitive respiration was due to a single respiratory pathway with a Ki for azide of 200 micrometer. A small amount of azide-sensitive respiration was detected in mitochondrial fractions obtained from chloramphenicol-treated cells, and it is likely that the azide-sensitive oxidase is localized in the mitochondrion. The determinants for the azide-sensitive and hydroxamate-sensitive oxidases segregate in a Mendelian manner in crosses and are either unlinked or not closely linked to each other.


Biological characteristics of a type I restriction-modification system in Staphylococcus aureus.

Two restriction-modification systems, S1 and S2, are present in Staphylococcus aureus RN450 (S. Iordanescu and M. Surdeanu, J. Gen. Microbiol., 96:277-281, 1976). System S2 affects phage multiplication after both infection and transfection. Unmodified plasmid and chromosomal DNAs are also not expressed following transduction and transformation into a restrictive host. Restricted phages are, however, capable of conferring phage-mediated competence, although the state of competence does not affect the restriction-modification system. The restricting activity of system S2 is inactivated by heat treatment of the cells. An enzymatic activity that restricts unmodified phage DNA in the presence of ATP, Mg2+, and S-adenosylmethionine was recovered from cell-free extracts of a strain RN450 derivative.


Effect of myosin DTNB light chain on the actin-myosin interaction in the presence of ATP.

The influence of the DTNB light chain of myosin on its enzymatic activities was examined by studying the superprecipitation of actomyosin and the actin-activated ATPase of heavy meromyosin (HMM) [EC 3.6.1.3]. Although the Ca2+-, Mg2+-, and EDTA-ATPase activities of control and DTNB myosin were practically the same, the superprecipitation of actomyosin prepared from actin and DTNB myosin occurred more slowly than that of control myosin. The apparent binding constant obtained from double-reciprocal plots of actin-activated ATPase of DTNB HMM was lower than that of control HMM. Recombination of DTNB myosin and HMM with DTNB light chains restored the original properties of myosin and HMM. The removal of DTNB light chain from myosin had no effect on the formation of the rigor complex between actin and myosin. These results suggest that the DTNB light chain participates in the interaction of myosin with actin in the presence of ATP.


The identification of sulphatides in human erythrocyte membrane and their relation to sodium-potassium dependent adenosine triphosphatase.

Sulphatides (ceramide galactose-3-sulphate) were isolated from human erythrocyte membranes. The amount obtained was 3.3 mg from 6.7 kg of wet cells, or 1.5 X 10(-9) mol per g dry cells. The polar part was shown to be galactose-3-sulphate by chromatographic analysis, infrared spectrometry, and mass spectrometry after solvolytic desulphation. The ceramide part consisted of three major molecular species, sphingosine-palmitic acid, sphingosine-2-hydroxypalmitic acid, and phytosphingosine-2-hydroxypalmitic acid, as shown by thin-layer chromatography, mass spectrometry of galactosylceramides after desulphation, and gas chromatography of components after hydrolysis. The composition differed from other human erythrocyte sphingolipids. Although the amount of sulphatides is very low for erythrocyte, the ratio of sulphatide concentration and Na+-K+-ATPase activity [EC 3.6.1.3] is similar to the situation found for several animal tissues with an increased level of Na+ transport. This finding is discussed in relation to a recent model of sulphatide function in a transport unit for Na+ and K+ (cofactor site model).


Adenosine triphosphatase activity and "thick filament" formation of chicken gizzard myosin in low salt media.

The ATPase activity of chicken gizzard myosin was studied by varying the KCl concentration in the reaction medium. The following was thus found: (a) A sharp depression of the activity occurred when the KCl concentration was reduced to less than 0.3 M, showing the minimum activity around 0.15 M KCl. (b) The activity depression was removed by addition of urea or bay papain-digestion, but not by addition of p-chloromercuribenzoate. (c) In the KCl concentration where the activity depression occurred, the ATPase reaction proceeded in two distinct phases; the activity was relatively high in the early phase of the reaction and declined into the later phase where the steady state reaction took place. (d) In the KCl concentrations higher than that particular concentration or in the presence of urea, the ATPase reaction proceeded in one phase. (e) The temperature dependence of the ATPase activity in the early phase was of an ordinary magnitude being approximately equal to that of the ATPase activity in 0.6 M KCl. In contrast, the temperature dependence of the activity in the later phase was unusually small. Gizzard myosin in various concentrations of KCl was also examined by measuring the turbidity and the light-scattering intensity, and by observation under an electron microscope. The following was thus found: (a) In the KCl concentration where the activity depression occurred, there was a stagnation in the turbidity decrease as the KCl concentration was gradually increased and also the formation of "thick filaments," each of which was approximately 0.6-0.9 micron in length and 20-30 nm in diameter with no central "bare zone." (b) Addition of ATP induced dissociation of the thick filaments, and the dissociation occurred during the early phase of the ATPaseeaction. (c) Moreover, the temperature dependence of the ATP-induced dissociation rate was approximately equal to that of the ATPase activity in the early phase. On the basis of the findings mentioned above, it is concluded that the activity depression results from the ATP-induced dissociation of myosin filaments. Moreover, since high concentrations of KCl or urea also caused dissociation of myosin filaments and yet did not produce the activity depression, it was strongly suggested that gizzard myosin in the ATP-dissociated form must be different from that in the urea- or KCl-dissociated form, probably in the physical state of some myosin aggregates which were not detectable by the physical methods we used. As a side-observation, gizzard myosin filaments formed in the presence of ADP were found to be unusually long (longer than 2 micron), and they looked very similar to the particular filaments of skeletal myosin that were reported, by Moos, to be formed in the absence of the C protein.


Isolation and composition of the subunits of spinach chloroplast coupling factor protein.

A more convenient method for preparing large amounts of spinach chloroplast coupling factor is described, in which centrifugation of the EDTA-extracted chloroplasts is replaced by batchwise adsorption on DEAE-cellulose followed by filtration through Miracloth. Methods have been developed to purify the subunits from coupling factor dissociated by sodium dodecyl sulfate, involving hydroxylapatite chromatography followed by gel filtration with the detergent still present. The amino acid composition of the subunits purified by these methods was determined, with some differences noted in values for cysteine, tyrosine, phenylalanine, and methionine compared to previously published values. The stoichiometry of the subunits was estimated as 2:2:1:1:2 from their relative adsorption of dye after gel electrophoresis, compared to dye adsorbed by known amounts of the purified subunits. Estimates of subunit stoichiometry are rounded off to nearest whole numbers; actual preparations of coupling factor usually show less than complete amounts of the two smallest subunits.


The epsilon subunit of Escherichia coli coupling factor 1 is required for its binding to the cytoplasmic membrane.

The coupling factor, F1-ATPase of Escherichia coli (ECF1) contains five different subunits, alpha, beta, gamma, delta, and epsilon. Properties of delta-deficient ECF1 have previously been described. F1-ATPase containing only the alpha, beta, and gamma subunits was prepared from E. coli by passage of delta-deficient ECF1 through an affinity column containing immobilized antibodies to the epsilon subunit. The delta, epsilon-deficient enzyme has normal ATPase activity but cannot bind to ECF1-depleted membrane vesicles. Both the delta and epsilon subunits are required for the binding of delta, epsilon-deficient ECF1 to membranes and the restoration of oxidative phosphorylation. Either delta or epsilon will bind to the deficient enzyme to form a four-subunit complex. Neither four-subunit enzyme binds to depleted membranes. The epsilon subunit, does, however, slightly improve the binding affinity between delta and delta-deficient enzyme suggesting a possible interaction between the two subunits. Neither subunit binds to trypsin-treated ECF1, which contains only the alpha and beta subunits. A role for gamma in the binding of epsilon to F1 is suggested. epsilon does not bind to ECF1-depleted membranes. Therefore, the in vitro reconstitution of depleted membranes requires an initial complex formation between epsilon and the rest of ECF1 prior to membrane attachment. Reconstitution experiments indicate that only one epsilon is required per functional ECF1 molecule.


Purification of the rep protein of Escherichia coli. An ATPase which separates duplex DNA strands in advance of replication.

The product of the rep gene of Escherichia coli catalytically separates phiX174 duplex DNA strands in advance of their replication, utilizing ATP in the process (Scott, J. F., Eisenberg, S., Bertsch, L. L., and Kornberg, A. (1977) Proc. Natl. Acad. Sci. U. S. A. 74, 193-197). The enzyme has now been purified to near-homogeneity. Relatively large quantities were obtained from ColE1-plasmid-containing cells in which the enzyme level was 7 to 10 times above wild type. The assay for rep protein was based on its essential role, with phage-induced cistron A protein, in enzymatic synthesis of phage phiX174 (+) strands, using duplex circular DNA as template. The protein exhibits a molecular weight of 65,000 under denaturing and reducing conditions. The turnover number of the enzyme is approximately 6800 ATP molecules/min in strand separation as measured by extent of replication, or in an uncoupled reaction using single-stranded DNA effector.


ATP utilization by rep protein in the catalytic separation of DNA strands at a replicating fork.

Hydrolysis of ATP by rep protein proceeds in the presence of a single-stranded region of DNA 4 residues long, but the true effector for rep ATPase appears to be a replicating fork rather than a random coil. At or near a fork in duplex DNA, rep ATPase action is different from what it is on DNA lacking secondary structure (single-stranded): (i) Km for ATP is lower, (ii) specificity is for ATP and dATP with no action on other nucleoside triphosphates, (iii) sensitivity to certain ATP analogs is reduced, (iv) presence of a DNA-nicking enzyme (e.g. cistron A protein induced by phiX174) is required, and (v) Escherichia coli DNA binding protein facilitates rather than inhibits. During the separation of strands accompanying replication, 2 molecules of nucleoside triphosphate (ATP or dATP) are hydrolyzed for every nucleotide polymerized. Utilization of ATP by rep protein may provide energy for catalytic strand separation at a fork in advance of replication.


Thymidylate synthetase and dihydrofolic acid reductase in the stimulated human lymphocyte.

The phytohemagglutinin (PHA)-stimulated human lymphocytes demonstrated trace or no activity (dihydro) folate acid reductase using three methods including a radioassay, but demonstrated ample activity of thymidylate synthetase. This was true regardless of the day of harvest, (first through seventh) of the stimulated lymphocyte. The lymphocyte extracts revealed no inhibitor to the reductase enzyme when these extracts were added before the liver extracts to the assay system. When methotrexate (MTX) was added to the culture media of the lymphocytes, there was, as expected, an increase in the synthetase activity, but the expected rise in  the reductase activity did not occur, it remained nil. On the other hand, MTX did influence the incorporation of nucleosides by the stimulated lymphocytes in a fashion similar to its action on the incorporation of the same nucleosides by other cells.


A suppressor T cell of the mixed lymphocyte reaction specific for the HLA-D region in man.

The mixed lymphocyte reaction (MLR) is the proliferative response of one individual's lymphocytes cultured in the presence of another individual's lymphocytes. In man, the MLR is elicited by cell surface antigens coded for by the HLA-D gene locus. This locus is among a cluster of genes which are located on the sixth chromosome and which include genes coding for the major histocompatibility antigens HLA-A, B, and C as well as HLA-D. If the stimulator cell possesses D locus antigens not present in the responder, the lymphocytes of the latter will undergo blast transformation resulting in DNA synthesis which can be measured. A vigorous response in the MLR to allogeneic cells is the rule among healthy individuals. We describe studies of a 23-yr-old man whose lymphocytes respond normally to mitogens and soluble antigens but fail to respond to allogeneic cells in the MLR. His medical history is unremarkable except that he received thymic irradiation as an infant. HLA typing revealed that he is homozygous for HLA-A2, B12, and Cw5 as well as for the D locus antigen Dw4. When his lymphocytes were added to the responder lymphocytes of other persons homozygous for the same HLA antigens, their responses to allogeneic cells but not mitogens were suppressed by 50-95%. Their responses to a soluble antigen, tetanus toxoid, were suppressed to a lesser degree. These inhibitory effects were mediated by a relatively radioresistant thymus-derived (T) lymphocyte. Further studies of the requirements for MLR suppression revealed that only persons heterozygous or homozygous for the Dw4 antigen were inhibited by the suppressor T cell. This effect was not altered by differences in the HLA-A, B, or C antigens between the suppressor and responder. It is concluded that genes in or near the HLA-D locus code not only for antigens (primarily on bone marrow-derived (B) cells), that elicit the MLR, but also for structures on T cells, or possibly macrophages, which are recognized by MLR suppressor T cells.


Effect of aminonucleoside nephrosis on immune complex localization in autologous immune complex nephropathy in rats.

The effect of increased capillary permeability on glomerular immune complex localization was studied in rats immunized with proximal tubular antigen (Fx1A) to induce autologous immune complex nephropathy (AICN). AICN rats were made proteinuric by injection or unilateral renal perfusion with aminonucleoside of puromycin (PA) before developing subepithelial complex deposits. Control AICN kidneys developed diffuse granular deposits of IgG and Fx1A on the subepithelial surface of the glomerular basement membrane (GBM) at 3 wk by immunofluorescence and electron microscopy, and deposits increased in subsequent weekly biopsies. In contrast, PA-nephrotic AICN kidneys developed few or no GBM deposits and a significant increase in mesangial localization of IgG and Fx1A during the period of PA-induced proteinuria. These alterations in complex localization were documented both in rats with PA nephrosis and in unilaterally PA-nephrotic kidneys compared with contralateral controls in the same animals, thus excluding any effect of PA on the immunopathogenetic mechanism in AICN as an explanation for these findings. The absence of GBM deposits closely correlated with reduced staining for polyanionic glomerular sialoprotein in proteinuric kidneys, since PA-perfused kidneys studied 2 wk after resolution of proteinuria demonstrated return of normal staining for sialoprotein and development of subepithelial complex deposits similar to those in contralateral control kidneys. These studies demonstrate that properties of the glomerulus itself play an important role in determining the site of complex deposition in experimental AICN and suggest that electrophysical characteristics of the glomerular capillary wall may influence complex localization on the GBM.


The use of the low back and the dorsal scales in the identification of functional low back patients.

Scores from the Low Back (Lb) and the Dorsal (DOR) scales of 20 patients with functional low back pain, 20 patients with functional gastrointestinal pain, and 20 psychoneurotic patients were compared. Among the various proposed cut-off scores, the Lb score of 11 yielded the highest rate (75%) of correct identification of Low Back patients and a hit rate similar to those reported in previous studies. However, it was noted that this 75% hit rate did not achieve statistical significance. Furthermore, the data indicated that neither the Lb nor the DOR scales could differentiate the Low Back patients from other psychosomatic and psychiatric patients and suggested that these two scales should be used with extreme caution by clinicians.


Personality characteristics of physically impaired adolescents.

Investigated MMPI profile differences between disabled and nondisabled (normals). Comparison of mean raw scores on the MMPI was made by t-tests. A preliminary analysis that compared adolescents with congenital versus traumatic impairment yielded only one significant finding: Scale O (Si) (p is less than .05) for females with congenital impairment. Subsequently, after the congenital and traumatic groups for each sex were combined, t-tests were computed on mean raw scores of the MMPI for each scale between the disabled and nondisabled adolescent groups. The profile patterns of the disabled groups for both sexes were similar; however, the male disabled adolescents had significantly higher scores on Scales 1, 2, 5, 8 and 9, while the disabled female adolescents had significantly higher scores on Scales F, 1, 6, 7, 8 and 9. Inspection of the profiles revealed minimal sex differences among the disabled groups. It was suggested that the common factor of moderate to severe chronic physical impairment may serve to reduce normative sex differences in personality development at least as reflected by the MMPI.


Kinetics of CTL induction by concanavalin A.

Induction of maximal CTL activity was achieved within 12 hr of exposure to Con A in vitro in various mouse lymphoid cell populations. These included spleen cells from normal unsensitized mice, spleen cells from mice previously immunized with alloantigen, and mouse spleen cells exposed to alloantigen in long-term mixed leukocyte culture (LTMLC). Although induction of maximal incorporation of tritiated thymidine was accomplished within this same period in the cells obtained from LTMLC, a much longer period of Con A exposure (greater than 24 hr) was required for freshly prepared spleen cells from normal or previously immunized mice. These findings indicate that the increased tritiated thymidine uptake induced in freshly prepared spleen cells on continued exposure to Con A beyond 12 hr is not associated with the development of cytolytic activity, and that it probably represents stimulation of subpopulations no longer present in the LTMLC population where positive selection for cells responsive to cellular alloantigens has taken place.