Multiple molecular forms of acetylcholinesterase from human erythrocyte membranes. Interconversion and subunit composition of forms separated by density gradient centrifugation in a zonal rotor. Quantitative separation of eight multiple molecular forms of acetylcholinesterase isolated from human erythrocyte membranes was accomplished by sucrose density gradient centrifugation in a zonal rotor. Hydrodynamic properties of the four most abundant oligomers were investigated by analytical ultracentrifugation. The S20, w values obtained for these components ranged from 12.5 S to 19.0 S with corresponding molecular weights between 500 000 and 1 131 000. These oligomers are built up of 6-14 subunits. Analysis of each isolated homogenous form after storage for 6 months at 4 degrees C revealed interconversion into the original eight components. Upon addition of Triton X-100 the oligomers disaggregate to yield a single 7-S form, which reaggregates upon removal of the detergent. Such reaggregation can be prevented by chaotropic ions indicating that hydrophobic interactions are important in the formation of these oligomeric forms.


Interpretation of the Mössbauer spectra of the four-iron ferredoxin from Bacillus stearothermophilus. The Mössbauer spectra of both oxidized and reduced ferredoxin from Bacillus stearothermophilus have been analysed using computer fits to theoretical spectra obtained from a spin Hamiltonian. A consistent set of parameters was obtained from fits to spectra obtained over a wide range of temperature and magnetic field. These results are interpreted in terms of a model for the active centre which is consistent with its electronic and magnetic properties in both redox states. In the model for the oxidized centre all four iron atoms have essentially the same valence, intermediate between ferric and ferrous, with one pair spin-up and the other pair spin-down. On reduction the extra electron goes predominantly to one pair of iron atoms which become ferrous with the other pair remaining substantially unchanged. Using this model it is possible to obtain relationships between the spin Hamiltonian parameters for individual iron atoms and those for the coupled centre. This can give further insight into the relation between the observed electron paramagnetic resonance and Mössbauer spectra.


A proton nuclear-magnetic-resonance study of self-stacking in purine and pyrimidine nucleosides and nucleotides. The concentration dependence of the chemical shifts of the protons H-2, H-8 and H-1' of ATP4- and of Mg(ATP)2- , of all non-labile protons of adenosine, of H-5, H-6 and H-1' of TUP, and of H-5, H-6, H-1', and H-2' of uridine have been measured. The results for the purine derivatives are consistent with the isodesmic model of indefinite non-cooperative stacking; for adenosine K = 15 +/- 2M-1, for ATP K = 1.3 +/- 0.2 M-1 and for Mg(ATP)2-K = 3.6 +/- 0.3 M-1. For the pyrimidines, uridine and TUP, stacking is much weaker and the stability constant could only be estimated; for uridine k is less than or equal to 0.5 M-1, and for UTP K approximately 0.3 M-1.


Immunochemical characterization of cyanogen bromide degradation products of M and N blood-group glycopeptides. The major glycopeptides purified from the tryptic digests of M and N blood-group glycoproteins were degraded with cyanogen bromide into two fragments. The chemical composition and serological activities of the fragments obtained were determined. The results show that M and N blood-group determinants are located on the smaller N-terminal fragments, containing 8 amino acid residues and only alkali-labile oligosaccharide chains. Two of 8 amino acid residues are different in M-specific and N-specific glycopeptide. All glycopeptides obtained inhibited Vicia graminea anti-N lectin, but the N-terminal fragment of N-glycopeptide was a better inhibitor than others. Treatment with neuraminidase or acetylation of amino groups destroyed the M and N blood-group activity and increased the activity towards Vicia graminea anti-N lectin of all glycopeptides studied.


Isolation and characterization of the messenger RNAs for mouse embryonic globin chains. Messenger RNAs for mouse embryonic globins were purified from yolk sac erythroid cells by oligodeoxythymidilate-cellulose chromatography and sucrose density centrifugation. Full-sized complementary DNA copies of embryonic globin mRNAs were synthesized. Acrylamide gel electrophoresis of these RNAs indicate an average molecular weight of 220 000, including a polyadenylated sequence of about 35 residues, as determined by hybridization to [3H]polyuridylate. The wheat-germ translation products of mRNAs have the size and the ionic characteristics of the four embryonic globin chains alpha, x, y and z. Hybridization kinetics in vast RNA excess were performed and compared to standard r0t curves of adult globin messengers, demonstrating a total base sequence complexity of about 880 000 daltons, that is four different RNA sequences of 220 000 molecular weight. The titration of embryonic globin cDNAs with increasing amounts of their complementary RNA templates indicates that the embryonic globin messengers were isolated at a high degree of purity.


Further studies on the quaternary structure of D-ribulose-1, 5-bisphosphate carboxylase from Alcaligenes eutrophus. Homogeneous D-ribulose-1,5-bisphosphate carboxylase, isolated from the hydrogen bacterium Alcaligenes eutrophus, has been studied by analytical ultracentrifugation. Sedimentation equilibrium experiments showed the enzyme to have a molecular weight M c =0 r =534000. The sedimentation coefficient was S0(20); w = 14.1S. The two types of subunits constituting the ribulosebisphosphate carboxylase were separated by gel filtration in the presence of sodium dodecylsulphate and the amino acid compositions of the isolated large and small subunits were determined. Rabbit antibodies were developed against the ribulosebisphosphate carboxylase and its isolated subunits. The specific reactivity of the respective antibodies with their homologous antigens was proven by double immunodiffusion and quantitative immunoprecipitation analyses. Antibodies elicited against the whole enzyme also reacted with both the isolated large and small subunits as did the subunit-specific antibodies with the whole enzyme. There was no immunological correspondence between the large and the small subunits. The specific inhibition of the enzyme activity by antibodies directed against sites on the large subunit suggests that the catalytic function resides in the large subunit. Electron microscopic examination of antibody . carboxylase complexes formed upon mixing of the specific immunoglobulins G with the enzyme was used to verify the arrangement of the large and small subunits in our recently proposed structural model of the enzyme molecule. The results confirmed that the large subunits are located in the central two layers of the four-layered enzyme molecule, whereas the two outer layers consist of small subunits. The observations are discussed with respect to an alternative model for the quaternary structure of ribulosebisphosphate carboxylase from tobacco.


Treatment of seven cases of chronic granulomatous disease with sulfamethoxazole-trimethoprim (SMX-TMP). Seven male Japanese children with chronic granulomatous disease were given sulfamethoxazole-trimethoprim (SMX-TMP) for recurrent pyogenic infections, most of which had proved difficult to control with other antibiotics. With continous treatment the children remained free of infections severe enough to necessitate hospitalization, except on one occasion. Serious complications, including hematological disorders, never occurred during therapy and there were no changes in leukocyte function during therapy. These results indicate that SMX-TMP should be considered in the treatment of bacterial infections associated with chronic granulomatous disease.


Urinary excretion of aromatic acids in hyperphenylalaninemic states: response to a protein challenge. A protein challenge (3g/kg) was given to 15 hyperphenylalaninemic patients for 4 days and urinary excretion of aromatic phenylalanine metabolites (e.g. phenylpyruvic acid; mandelic acid; ortho-hydroxy-phenylacetic acid) was studied by quantitative gas chromatographic analysis. Two groups of patients, different in their serum phenylalanine pattern, could be distinguished from the pattern of urinary aromatic acid excretion. In the first group, the aromatic acid excretion declined towards the end of the challenge whereas aromatic acid excretion increased throughout the challenge in the second group. The differences could be ascertained by statistical analysis.


Hyperglycinuria with nephrolithiasis.

Hyperglycinuria with nephrolithiasis. The case of a seven and a half-year-old girl with hyperglycinuria, oxalate nephrolithiasis, and a normal plasma amino acid pattern is presented. Hyperglycinuria amounted to 400 mg of glycine in 24 h urine and the stone was composed of calcium oxalate dihydrate. The metabolic relationship between glycine and oxalate is discussed. It is possible that the association of nephrolithiasis and hyperglycinuria was coincidental, although the case of familial hyperglycinuria with nephrolithiasis reported by De Vries and collaborators and our case suggest the possibility of a relationship between the aforesaid compounds in vivo.


Combination chemotherapy and radiotherapy for Hodgkin's disease in children. 31 children with Hodgkin's disease were treated at the Children's Hospital, University of Munich, between 1963-1976. Before 1971 the diagnosis of abdominal involvement was based solely on physical findings and radiological studies. Treatment consisted of involved-field radiation; chemotherapy was added in selected cases. Of 15 children treated between 1963-1971, 5 are still in their first remission and 2 in their second remission. Since 1971 exploratory laparotomy and splenectomy have been performed. Radiation therapy was changed from involved- to extended-field therapy and combination chemotherapy was added in all the children. No patient from this group of 16 children relapsed during a median observation time of 24 months (range 12-55 months). The toxicity of the combined treatment does not seem to be prohibitive.


Femoral hypoplasia-unusual facies syndrome, from another viewpoint. A female infant with the "femoral hypoplasia-unusual facies" syndrome is presented. Most of the findings observed in this child have also been described in the "caudal regression syndrome". The similarity and probable identity between these two syndromes is discussed on the basis of our patient and others from the literature.


Multiple skeletal anomalies in the "13q-" syndrome. A patient with the "13q-" syndrome is reported. The typical association of congenital malformations was found. Multiple and unusual skeletal anomalies included absent thumbs, club-feet, coxa vara, diastasis of the pubic symphisis and extensive spina bifida occulta. These appear to be part of the multiple system involvement due to the chromosomal deletion. Chromosome analysis is indicated in patients with multiple skeletal anomalies, especially if the thumbs and radial axis are involved.


Endoscopy in pediatric gastroenterology. Seventy two endoscopies of the upper gastrointestinal tract and one hundred and fifteen colonoscopies were performed in children aged nine months to seventeen years. The experience gained with different types of endoscopes suggests that endoscopy of the upper gastrointestinal tract should be only performed with pediatric instruments. These are: GIF-P2(1) (max. outer diameter 9 mm) and TX-7(2) (max. outer diameter 10mm). For colonoscopy, the ideal pediatric instrument is not yet available. Sigmoidocoloscopes like the CFMB1 (length 1115 mm) and the F-91-S2 (length 650 mm) are suitable for older children.


Preliminary immunological studies in spinal muscular atrophy. Evidence of impaired cell-mediated immunity in children with spinal muscular atrophy was obtained using lymphocyte transformation with PHA (20 children) and skin tests with tuberculin and DNCB (35 children). Laryngological examination of 16 children demonstrated hypoplasia of lymphatic tissue in Waldeyer's ring and the cervical lymph nodes. An ectodermal defect involving both spinal cord and thymus is suggested.


Benign course of juvenile laryngeal papillomata. A 17 month-old child with laryngeal papillomata is presented. The symptoms were stridor and gradually developing dyspnea. He underwent two surgical procedures within 4 months, using microsurgical techniques, and eventually recovered. The etiological factors of this rare entity are discussed. The relatively benign course in our patient is stressed, in view of the usual protracted course which is reflected in the literature.


Spondylocostal dysostosis and urinary tract anomaly: definition and review of an entity. A two-year-old girl with spondylocostal dysostosis and urinary tract anomalies is described. Genetic counselling to the healthy non-consanguineous parents is based on the hypothesis of autosomal recessive inheritance. At birth this type of spondylocostal dysostosis can be differentiated radiographically from related conditions by the severe and generalized anomaly of the entire vertebral column, and by the typical configuration of the posteriorly-fused ribs. Only the cardinal clinical features of a short trunk and a short, immobile neck are consistently present in all patients. In one group of patients the severe thoracic malformation and lordosis contribute to cardiopulmonary problems with a fatal outcome in early infancy. The roentgenographic changes are less marked in other patients, whose subsequent clinical course is usually without major problems but remains at present insufficiently known.


Glucocorticosteroids and growth hormone secretion under physiological conditions and in states of steroid excess. Cortisol and growth hormone (GH) secretion (spontaneous variations at night and the release induced by insulin hypoglycaemia) were investigated in 69 children and adolescents. Statistical analysis of approximately 600 pairs of cortisol and GH values in this study demonstrated that physiological fluctuations of cortisol do not alter GH secretion. A review of the literature shows that GH secretion is consistently depressed in Cushing's disease of central origin and in Cushing's syndrome due to adrenal carcinoma. When acutely administered, doses higher than 100 mg of cortisol (or equivalent amounts of other steroids) per adult are necessary to block GH secretion and the hormones have to be given several hours previously. In long-term steroid treatment, suppression of GH is observed in only 1 out of 3 patients. The effect apparently does not persist beyond elimination of the last dose, i.e. generally not longer than 12 to 24 h. These data can be taken as a rationale for intermittent or alternating dosage schedules, and for the use of short acting derivatives if long-term, high-dose steroid treatment is necessary in children. It remains to be established whether growth deficiency in exogenous hypercortisolism is due to suppression of GH secretion, decreased production of somatomedins, direct antagonism of the action of somatomedins on growing cartilage, or a combination of these mechanisms.


Renal magnesium wasting, incomplete tubular acidosis, hypercalciuria and nephrocalcinosis in siblings. Polyuria, hyposthenuria, hypomagnesemia, hypercalciuria, advanced nephrocalcinosis, low citrate excretion and low glomerular filtration rates were observed in two female siblings who were followed over 10 years. Acid loading revealed incomplete distal tubular acidosis. Hypomagnesemia was due to renal magnesium wasting. It is suggested that the defect in tubular transport of magnesium is an important factor in the pathogenesis of this syndrome.


Sleep and aminophylline treatment of apnea in preterm infants. The influence of short-term aminophylline treatment on sleep behaviour was studied in six preterms infants with recurrent apnea. The incidence of apnea, respiratory pauses, and bradycardias which were closely related to the phases of active sleep, decreased during aminophylline treatment. However, the amount of active sleep remained unaffected. The mode of action of aminophylline is discussed in view of the previously proposed neurophysiological concept of apnea of prematurity.


Postmortem tissue and plasma concentrations of digoxin in newborns and infants. Postmortem tissue and plasma concentrations of digoxin were studied in 13 premature newborns, 6 mature newborns, and 5 older infants (age 1 to 14 months). The pertinent results of our study are as follows: The tissue digoxin concentrations tend to be higher in premature and mature newborns than in infants. This difference is statisitcally significant with respect to the concentration in myocardium and skeletal muscle. The renal digoxin concentration of premature newborns is significantly lower than that of mature newborns, the tissue concentrations in the other organs examined being essentially equal. In all age groups examined, skeletal muscle contains the greatest portion of digoxin, followed by the liver. The relation of myocardial to plasma digoxin concentration shows no significant difference between the various age groups. Within the groups, the variation is relatively large.


The effect of 5-hydroxytryptamine and epinephrine on newborn platelets. Blood levels of 5-hydroxytryptamine (5-HT) and epinephrine are increased in conditions such as severe hypoxia, and hypovolemic or septic shock. The effect of these agents, which influence the blood coagulation system, were studied in vitro on platelets from newborns. 5-HT was determined in platelets from 18 newborns and 28 adults. The 5-HT content was 0.305 +/- 0.144 microgram/10(9) platelets in newborns and 0.675 +/- 0.212 microgram/10(9) platelets in the adult controls, a difference which is statistically significant (P less than 0.001). The 5-HT content of newborn platelets proved to be higher than that reported in the literature. Further studies on platelet aggregation due to 5-HT and epinephrine were then carried out. These demonstrated first-phase aggregation in 40% of cases when both 5-HT and epinephrine were applied simultaneously, while in the majority of the newborns no reaction was encountered when each of these substances was applied separately. However, in 2 cases, simultaneous application of 5-HT and epinephrine produced both first- and second-phase platelet aggregation. The reduced response of the platelets in the newborn to these aggregating agents can be considered as a protection against disseminated intravascular coagulation (DIC), particularly in high-risk situations.


Clinical and aortographic assessment of the complications of arterial catheterization. Catheterization of the umbilical artery for the treatment of critically ill neonates provides a convenient method for monitoring blood gas tension and chemistry. The most important complications are thrombotic. Thirty eight aortographs were carried out in infants who underwent umbilical artery catheterization. 17/38 of the aortographs were pathological. Bacterial cultures were positive in 11/17, but only 4 coincided with pathological aortographs. Clinical signs indicating complications due to the presence of the catheter wwere observed in 10 cases. Post-mortem examination of eight subjects--three of whom had pathological aortographs--during the course of the investigation revealed only one case of thrombosis. This baby was considered to have died as a direct result of a thrombotic complication. In our experience the clinical signs of vascular complications and evaluation of the peripheral circulation in the ipsilateral leg remain the most important ways of assessing the indication for catheter-withdrawal.


Interrelation of maternal and foetal glucose and free fatty acids. The role of insulin and glucagon. At birth there is a statistically significant correlation between the maternal blood sugar and the umbilical venous and arterial blood glucose. The glucose concentration in the mother is higher than in the umbilical vein, and in the latter higher than in the umbilical artery. The free fatty acids also show a falling gradient from the mother to the umbilical vein and artery. This supports the assumption that glucose and FFA pass the placenta from the mother to the foetus. Insulin concentration is significantly higher in the mother than in the cord blood. There is however no difference between umbilical venous and arterial plasma insulin. A statistically significant correlation between glucose and insulin is only demonstrable in the maternal blood. Glucagon levels are about the same in maternal and umbilical arterial blood, but are significantly higher in umbilical venous blood. These results are consistent with the supposition that insulin and glucagon do not cross the placenta to a significant extent and can be synthesised by the foetus. In the first few hours after birth, a fall of glucose and a rise in FFA occur at the same time. Insulin tends to be lower than at birth and glucagon rises.


Excretion of 2-methyl-3-oxovaleric acid in propionic acidemia. A new case of propionic acidemia is presented, paying special attention to the early symptoms of this disease, such as increased drowsiness, muscular hypotonia, poor feeding, hypothermia, metabolic acidosis, ketonuria and vomiting. Investigation by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) revealed the excretion of fairly high amounts of 2-methyl-3-oxovaleric acid, a condensation product of two molecules of propionyl-CoA, as well as the known pathological metabolites such as propionic, 3-hydroxypropionic and methylcitric acids. Among the post mortem findings the histological studies of the liver were the most remarkable.


Recurrent aphasia with subclinical bioelectric status epilepticus during sleep. We present a six years follow up of a 6 1/2 year-old boy with recurrent aphasia, sporadic emotional regression and a convulsive disorder. The electroencephalogram reading during sleep showed continuous, generalized, hypersynchronous activity without clinical evidence of seizure ("subclinical bioelectric status epilepticus"). Clinical, laboratory and instrumental data revealed no evidence of a morphological lesion of the brain, either space-occupying, inflammatory, degenerative, or autoimmune. Remissions occured with and without anticonvulsant and steroid therapy. The hypothetical location, type, and etiology of the process are discussed. The syndrome appears to represent a functional disorganisation of speech-controlling areas of the brain.


The selenium state of children. II. Selenium content of serum, whole blood, hair and the activity of erythrocyte glutathione peroxidase in dietetically treated patients with phenylketonuria and maple-syrup-urine disease. The selenium content of serum, whole blood and hair was measured by neutron activation analysis in dietetically treated patients with phenylketonuria (PKU) and maple-syrup-urine disease (MSUD). Follow-up studies showed a decrease of the serum selenium content and the glutathione peroxidase activity of erythrocytes--a selenoenzyme--from normal values at the beginning of the diet to 20% (selenium) and 50% (gluthione peroxidase) of normal within 10--12 weeks of dietary treatment. In 36 patients the serum selenium content was lower at 6.7--28 X 10(-9) g/ml, independent of the age of the patients (0.5 to 10 years). The selenium content of whole blood was reduced: median = 98 X 10(-9) g/g dry weight; range 75 to 165 X 10(-9) g/g dry weitht (healthy children: median = 381 X 10(-9) g/g dry weight; range 245 to 588 X 10(-9) g/g dry weight). The selenium content of hair was markedly lower in the patients (median = 62 X 10(-9) g/g; range 13--140 X 10(-9) g/g) than in healthy children (median = 429; range 213 to 720 X 10(-9) g/g). The mean glutathione peroxidase activity of erythrocytes was reduced to 4.6 +/- 0.64 U37/g Hb, comparison to normal values (mean = 8.8 +/- 0.88 U37/g Hb).


Argininosuccinic aciduria: metabolic studies and effects of treatment with keto-analogues of essential amino acids. A 22 years old female with the late-onset of argininosuccinic aciduria was successfully treated for 5 months with a mixture of essential amino acids and their keto-analogues. There was a marked change in plasma ammonia, plasma amino acids and argininosuccinic acid excretion. A long term anabolic response was reflected by an increase of total serum proteins and serum albumin by about 1 g/dl during the first 5 months of treatment. There was a striking improvement in the patient's seizure disorder following institution of the keto-analogue therapy.


Cry analysis in infants with severe malnutrition. Sound spectrographic investigations of the cries of 5 infants, age 7 m to 2 y, with severe malnutrition (one with kwashiorkor and four with marasmus) were compared with the cries of 15 healthy children of corresponding age. The cry of the child with kwashiorkor resembled those of the normal infants. The cries of the marasmic children showed a significant increase in the minimum and maximum pitch, and the occurrence of biphonation and flat melody types. These features have also been found in the cries of children with brain damage. We therefore believe that cry and analysis can be an additional means of investigating to what degree the brain is affected in children with malnutrition.


Structure and closure mechanism of the human umbilical artery. The structure of the fully-patent umbical artery and rearrangement of its structural elements with postnatal closure were examined 10 centimeter long umbilical cord segments which were double-clamped at different time intervals after delivery. The fully-patent umbilical artery consists of two main layers: an outer layer of circularly arrange smooth muscle cells and an inner layer which shows rather irregularly and loosely arranged cells embedded in abundant metachromatic ground substance. No predominantly longitudinal arrangements of cells and fibers reported by earlier investigators could be identified in the inner layer. Closure of the umbilical arteries is initiated by numerous localized contractions which are mainly formed by muscle cells of the outer circular layer. Ultimate closure of larger segments of the umbilical arteries is also mainly produced by contractions of the outer layer. In contrast, the inner layer (which is rich in ground substance) seems to serve mainly as a plastic tissue which can easily be shifted in an axial direction and then folded into the narrowing lumen to complete closure. Electron microscopy reveals that the cells of this layer represent rather poorly differentiated smooth muscle cells which contain only a few tiny myofilaments and can therefore hardly contribute actively to the process of closure.


Attempted dietary treatment of a boy with hyperammonemia due to ornithine transferase deficiency. Dietary treatment of a male patient suffering from the delayed-onset type of OCT deficiency was attempted. Control of the hyperammonemia was attempted by restriction of protein intake, guided by monitoring the plasma ammonia and regular checking of the serum amino acid levels. The influence of supplementary citric acid or lactulose therapy on the plasma ammonia level was investigated and found to be negligible. The therapeutic effect of supplying ornithine and arginine (an essential amino acid in urea cycle disorders) is described. Despite intensive dietary treatment over two and a half years, a incorrigible hyperammonemic crisis resulted in the sudden death of our patient.


A biochemical basis for grouping of patients with urolithiasis. The urinary excretion of calcium, magnesium, oxalate, creatinine, phosphate and urate was investigated in patients with urolithiasis and in normal subjects. The excretion of oxalate and urate per mole creatinine and the quotients calcium/magnesium, calcium X oxalate/magnesium and calcium X oxalate/(magnesium X creatinine) were significantly higher in stone formers than in normal subjects. The mean creatinine-correlated urinary excretion of calcium was higher and of magnesium lower in patients with urolithiasis, but the differences were statistically not significant. The urine investigation was supplemented with analysis of calcium, magnesium, creatinine, urate, bicarbonate and chloride in serum and a qualitative analysis of stone composition. A simple schedule for a biochemical grouping of patients with urolithiasis is presented and on the basis of the analytical findings it was possible to classify 67% of patients with so-called 'idiopathic stone disease' according to these principles.


Active specific immunotherapy of advanced renal-cell carcinoma. The results of a specific active immunotherapy method following palliative nephrectomy in 31 patients suffering from advanced renal-cell carcinoma are presented. The control patient group comprised of 23 comparable cases, who did not receive immunotherapy. The minimum follow-up time for any patient is 4 years. The statistical 5-year survival rate of the immunotherapy group is 23.6% (SE = 7.8). 7 patients are still alive. The 5-year survival rate of the control group is 4.3% (SE = 4.3), without a single survival. The difference is statistically highly significant (p less than 0.001). No side effects were noted from this form of immunotherapy.


Diagnostic and therapeutic difficulties of leucoplakia of the renal pelvis. 2 cases of leucoplakia of the renal pelvis are reported. In the first one the diagnosis was established after nephrectomy and histological examination of the specimen. In the second one it was suspected on urine examination and was confirmed on histology. The current aspects about its cause and the difficulties concerning its diagnosis and treatment are discussed.


Treatment of renal tuberculosis. II. Microbiological study of 271 renal tuberculous cavities. 271 renal cavities from 94 nephrectomies were studied with fluorescence stain and Lowenstein-Jensen medium cultures. The degree of the sterilization of the lesions is demonstrated by the existence of Koch's bacilli into the cavities. The latter have been classified in different development stages according to their contents and the histological aspect of the wall. The results of this study showed that a reduced healing time allows a better conservation of the parenchyma. The in vivo rifampicin + isoniazid + ethambutol association produces the best and quickest stabilization of the cavities.


Eosinophilic cystitis.

Eosinophilic cystitis. 4 cases of eosinophilic granulomatous cystitis are reported. The clinical presentation of this rare entity is disucssed. It is suggested that the lesion is probably akin to eosinophilic enteritis or eosinophilic granuloma of the gastrointestinal tract and represents a local intense allergic reaction of eosinophilic type. The causative agents are yet to be identified. Management by an allergist is suggested as a first line of treatment as it appears that the bladder lesions are usually too extensive for local resection.


Retrocaval ureter.

Retrocaval ureter. 3 cases of retrocaval ureter (one of them presenting multiple urological anomalies) are discussed. In the presence of hydronephrosis, early operative reposition of the ureter is essential. Provided that the condition is diagnosed before extreme renal deterioration, the surgical repair can salvage remaining renal function.


Percutaneous nephrostomy. Aspects on applications and technique. Percutaneous nephrostomy was performed in 32 patients: in 8 patients for permanent and in 24 for temporary drainage. The most common indication was obstruction of the distal ureter due to malignancy. Puncture of the renal pelvis was performed with the aid of fluoroscopy and/or ultrasound. Change to larger catheters was achieved using the Seldinger technique. Complications in the form of bleeding occurred in 5 patients and were in conjunction with repuncture after the initial catheter had slipped out, or occured in connection with change to larger catheters. With the routine use of ultrasound and specially designed catheters the method should be an effective alternative to operative nephrostomy also in the nonemergency patient.


Analysis of urinary calculi formed by Nigerians. 50 stones formed by Nigerians were analysed using a combination of qualitative chemical analysis, and petrographic analysis with the Reichert Model NR 333 254, polarization microscope. The Bausch and Lomb binocular dissecting microscope was used for macroscopic examination of the specimens and for taking samples from the 'nucleus', inner and outer surfaces of the stones, for analysis. The results showed that calcium phosphate and magnesium ammonium phosphate accounted for 64% of the stones, calcium oxalate for 28% while uric acid accounted for 8%. This is in contrast to modern times Caucasian stones which are predominantly calcium oxalate. There appears to be a greater tendency for Nigerians to form large staghorn calculi than small stones that can passed spontaneously. The phosphate stones are known to be related to infection in the urinary tract. The findings on the composition of these calculi correlate with the previous findings of a high incidence of obstruction and infection in Nigerian patients with urinary calculi.


Air cystometry at different flow rates.

Air cystometry at different flow rates. Air cystometry was performed with 3 different flow rates in patients suffering from different bladder diseases. To demonstrate neurogenic bladder disorder a flow rate of 75 ml/min was recommended.


Growth retardation of calcium oxalate by sodium copper chlorophyllin. Earlier reports from our laboratories described the inhibitory effect of Mg++ and pyrophosphate on the dissolution and growth of calcium oxalate. In this report, growth of Ca oxalate in presence of very low concentration of chlorophyllin was followed by the Coulter counter and a multichannel analyser. The effect of chlorophyllin concentration on crystal formation was studied at different levels of supersaturation. A concentration of 0.1 microgram/ml produced a remarkable retardation of the induction period and the growth rate. The findings obtained in this study are discussed in the light of the crystal poisoning theory. Comparison was made between chlorophyllin and other growth inhibitors.


Balloon catheter dilatation of renal artery stenosis in the dog. A preliminary report. Renal artery stenosis was successfully created in 2 out 3 dogs using a nonsurgical, percutaneous and intra-arterial technique. In both instances, it was possible to reduce the stenosis by dilatation with a balloon catheter. The possible clinical implications of this technique are briefly discussed.


Recto-urethral fistula. Treatment by abdomino-perineal pull-through resection of the rectum. 4 patients with difficult and longlasting recto-urethral fistulas were treated by abdominoperineal pull-through resection of the rectum. Primary closure of the fistula occurred in all cases. We suggest that this operative technique should be taken into consideration in cases of difficult rectourthral fistulas.


Intrarenal teratoma in a newborn child.

Intrarenal teratoma in a newborn child. A case of intrarenal teratoma in a newborn child is reported. The scarcity as well as the clinical and radiological signs of this lesion are emphasized and the difficulties to distinguish it from a nephroblastoma are analyzed. Solid intrarenal tumours in the infant and the newborn child are in nearly all cases nephroblastomas. All other types of solid tumours (benign or malignant) are exceptional. This induces us to report the present case.


Successful surgical repair of solitus, dextrocardia, atrioventricular discordance, and double outlet right ventricle with l-malposition of the aorta. The authors report a rare case of situs solitus of viscera and atria, dextrocardia, atrioventricular discordance, atrial and ventricular septal defects, origin of both great arteries from the morphologic right ventricle, pulmonary valvar and subvalvar stenosis and l-malposition of the aorta which has been correctly diagnosed and succesfully operated on. The repair was accomplished by closing the ventricular septal defect through a ventriculotomy in the systemic venous ventricle (morphologic left), and placing a conduit between this ventricle and the pulmonary artery which had been ligated proximally. The conduit was a fresh segment of homograft valved ascending aorta fixed on a Dacron woven tube. The authors discuss: firstly the reasons for performing this type of repair instead of the intraventricular one recommended by the Mayo Clinic group in the past and secondly the peculiar location of the conducting tissue and the method of avoiding it when approaching the ventricular septal defect.


The effects of the d- and l-optical isomers of propranolol on the ventricular fibrillation threshold. The minimum current required to induce ventricular fibrillation was determined in 22 sodium pentobarbital anesthetized open chest dogs in the presence of the d- or l-isomers of propranolol at concentrations of 0.2, 0.5, and 1.0 mg/kg. The hearts were paced from the right atrium at a constant basic cycle length of 350 msec and the current was delivered during the vulnerable period to the right ventricle as a 120 msec train of 4 msec square wave pulses at 100 Hz. The l-isomer of propranolol increased the ventricular fibrillation threshold (VFT) above control values at all concentrations; the maximum increase ranged from 17.8 to greater than 420.8% change from control. In contrast, the d-isomer caused only a small increase in the ventricular fibrillation threshold with the maximum increase ranging between 14.0 and 65.2% change from control. The maximum increase in VFT for the racemic mixture of propranolol was 44.7 to greater than 500.0% above control and was similar to the l-isomer alone. Since the beta-blocking potency of the l-isomer is 100 times the potency of the d-isomer we conclude that propranolol's action in increasing the ventricular fibrillation threshold is dependent on its beta-blocking effect.


Cardiac filling pressures in acute inferior myocardial infarction with and without right ventricular involvement. Right heart and pulmonary artery pressures were measured in 28 patients with inferior acute myocardial infarction, 12 of whom also had ECG evidence of right ventricular involvement (RVI). Those with RVI had significantly higher mean right-sided filling pressures 9.3 mm Hg, SD +/- 4.5 than those without RVI, 4.3 mm Hg, SD +/- 1.9. A steeper relationship between right ventricular enddiastolic and mean pulmonary artery pressures was seen in patients with RVI in contrast to those without, where no marked rise in right-sided filling pressures with increasing mean pulmonary artery pressures was seen.


Right and left ventricular flow during mechanical alternans. Pulmonary and aortic blood flow simultaneous with right and left ventricular isometric tension were measured in anesthetized dogs during mechanical alternans induced by rapid atrial driving. It was found that any steady state of mechanical alternation can be altered to another steady state by a single premature or delayed beat. The relationship between the force of the alternating beats was not necessarily equal in the two ventricles. Pulmonary and aortic flow changed markedly with changing steady states of mechanical alternation, at the same heart rate, within wide range of flow levels. Moreover, inequality between the mechanical alternation of the two ventricles was at times associated with disparity of aortic and pulmonic flow. The possible mechanism and clinical significance have been mentioned.


The recurrence risk in congenital heart disease. We have studied 1148 'isolated" cases of congenital heart disease, taken from a general series of 1255 cases. The recurrence risk ranged from 0.9% for relatives of male cases of VSD and for female cases of aortic stenosis to 10.5% for relatives of female cases of atrioventricularis communis; mean value 2.5%. Heritability ranged from 50% to 95%, mean 65%. Full concordance was more frequent in 1st-degree relatives of Fallot's tetralogy patients, while discordance was common in more distant relatives. We propose a hypothesis of several overlapping polygenic systems for the causation of CHD.


Computer prediction of left ventricular complicance throughout diastole in normal patients. This study deals with the development of a computer program to predict instantaneous left ventricular complicance, as defined by the tangent modulus E, throughout diastole. Diastole is divided into discrete time intervals according to the major events which occur: the start of isovolumic relaxation (aortic valve closure), mitral valve opening, the point of minimum left ventricular pressure, the junction of the rapid and slow filling phases, the start of atrial systole, and the peak of the 'a' wave. Each interval is separated into subintervals. Over each subinterval two mechanisms are assumed to operate: myocardial relaxation or contraction producing a pressure change without an accompanying volume change, followed by explansion of the left ventricle at constant pressure. Although these mechanisms occur simultaneously in the intact heart, they are treated sequentially in a multistage computer program that employs the finite element technique to determine the displacements within a thick-walled ellipsoidal shell. The smaller the time interval between successive stages, the closer is the approximation to the actual continous process of myocardial relaxation, contraction, and distension. Diastolic determinants revealed in this investigation are the mechanical properties of the myocardium, the state variables of pressure and volume, and the control variables of wall thickness and cavity size. In isovolumic relaxation, the myocardium relaxes and the ventricular wall thickens to reduce intracavitary pressure. The relaxation process continues and intraventricular pressure falls to a minimum (0-point) while ventricular volume increases after mitral valve opening. In the succeeding phases, excluding atrial systole, ventricular filling pursues, the properties of the myocardium change, there is an increase in tone (possibly due to myocardial contraction), the wall thins and intraventricular pressure rises. Computer prediction shows that at the start of diastole the tangent modulus is approximately 6 times the enddiastolic value, and is nearly 0 at the onset of the slow filling phase. Tangant modulus is a useful index by which to distinguish normal from abnormal patients provided the characteristics of E as a function of time are recognized and compared throughout diastole.


The influence of acute alterations in coronary blood flow on left ventricular diastolic compliance and wall thickness. The extent to which alterations in the coronary blood flow influence left ventricular (LV) diastolic compliance and wall thickness was studied in isolated, isovolumic (balloon in LV), blood perfused dog hearts. Studies were performed at a ventricular volume which was associated with an LV systolic pressure of 100 mm Hg and changes in LV diastolic pressure (at this constant volume) were taken as changes in LV diastolic complicance. When coronary perfusion pressure was reduced to zero (5 min of global ischemia), LV diastolic wall thickness fell from 9.9 +/- 0.9 mm to 9.2 +/- 0.7 mm (P less than 0.01) and LV diastolic pressure fell only 1.3 mm Hg (from 5.5 +/- 1.0 mm Hg to 4.2 +/- 1.1 mm Hg P less than 0.01). In contrast to these findings in 'near normal' ventricles, global ischemia in severely injured hearts (with unphysiologically high diastolic pressures) resulted in marked reductions in diastolic pressure. Thus, moment to moment changes in coronary blood flow can result in alterations in the diastolic properties of the LV. However, these are extreme interventions, and in the clinical setting, changes in coronary dynamics are unlikely to cause significant changes in LV diastolic pressure.


Plasma digoxin levels in infants, children and adults, using a radioimmunoassay micromethod. The radioimmunoassay method for estimation of plasma digoxin concentration was modified to use samples of 0.1 ml plasma for use in infants and children. The optimal daily maintenance dose for neonates, infants, and children was found to be between 0.01 and 0.015 mg/kg. Despite high plasma digoxin concentration (3-4 ng/ml) in some infants and children, no evidence of toxicity was found on electrocardiogram. 3 patients with accidental overdose, however, had plasma digoxin levels above 5 ng/ml and 2 of these had atrioventricular block.


The ascending slope of the apexcardiogram. Its relationship with angiographic determination of left ventricular function. The relationship between the ascending slope of the apexcardiogram (ACG) and direct determination of left ventricular (LV) performance as assessed angiographically by measurement of the ejection fraction (EF), mean velocity of circumferential fiber shortening (mVcF), mean normalized systolic ejection rate (MNSER), and percentage of the systolic shortening of ventricular long axis (%L), were studied in 40 patients with a wide variety of cardiac diseases. The ascending slope correlated closely and significantly (P less than 0.05) with all of them: (1) EF (r = 0.774), (2) mVcF (r = 0.776), (3) MNSER (r = 0.767) and (4) %L (r = 0.668). In a control group of 10 normal subjects, phi was determined and compared with that obtained in patients with depressed LV function, the difference being statistically significant (P less than 0.01). Other noninvasive indices derived from simultaneous phono-mechano-cardiographic readings were also studied. The calibrated ACG did not correlate with any of the hemodynamic indices; however the calibrated carotidogram did correlate with the peak systolic aortic pressure (r = 0.503). The ratio preejection period/left ventricular ejection time correlated significantly, but less than previously reported with the EF (r = 0.574). We conclude that the value of phi leads to additional evidence supporting the use of quantitative ACG as a noninvasive measure of LV performance in patients with cardiac disease.


A mechanism for shifts in the diastolic, left ventricular, pressure-volume curve: the role of the pericardium. Attempts to explain acute shifts in the pressure-volume relationship by changes in myocardial stiffness have been unsupported by animal experimentation and discouraged by calculations showing that making half the ventricle rigid would hardly account for the clinically observed upward shifts in the curve. Data collected in open-chest anesthetized dogs during volume loading with the pericardium open and closed indicate that when the pericardium is closed, left ventricular enddiastolic pressure is better predicted by right ventricular pressure than by left ventricular dimensions. These data support the hypothesis that acute, upward shift in the pressure-volume curve may be caused by an increase in pericardial pressure, in turn caused by an increase in the volume of the intrapericardial contents. This follows from the fact that measured left ventricular diastolic pressure is equal to the sum of the pressure differences accross the myocardium and the pericardium. Thus, increases in pericardial pressure raise measured ventricular diastolic pressure without change in ventricular volume: hence, an upward shift in the pressure-volume curve.


Control of the frog heart relaxation by Na-Ca exchange. Frog heart relaxation was analysed under voltage clamp conditions as the tension decay observed after membrane potential had been stepped back to its resting value. It appeared mostly exponential with a time constant of about 190 msec. Relaxation rate decreased with the external Na concentration. It fell to 1/10 in a Na-free solution. Relaxation rate decreased also following an increase in the intracellular Na content by application of veratrine. When switching from a high to a low Na-containing solution the relaxation rate reached rapidly a value transiently higher than the steady-state's (and vice versa). In Na-free solution, adrenaline accelerated tension decay, an effect not noticeable on frog heart in Ringer solution. In frog myocardium, part of the intracellular Ca concentration decrease which occurs during each cardiac cycle directly results from a calcium extrusion carried out by the Na-Ca exchange. Such a Ca decrease, besides internal Ca storage, should play a primary role in the relaxation of contractile tissues whose cells have a large surface/volume ratio.


Left ventricular compliance in acute transmural myocardial infarction in man. 130 patients with recent transmural myocardial infarction were studied in order to evaluate changes in left ventricular compliance. Left heart catheterization and cineangiographic left ventriculography were performed. The modulus of chamber stiffness (K) was determined from the slope of the linear relation between volume stiffness (dP/dv) and pressure; the modulus of muscle stiffness was evaluated by the Laird index (asymptotic slope of logP vs log V). Patients were divided in 3 groups: Group A included 5 patients studied before and after myocardial infarction who experienced an increase in the modulus of chamber and muscle stiffness after infarction. Group B included 10 patients who were studied at the onset of infarct and 15 days later: this group demonstrated changes in LV compliance with, most often a rightwardshift of P-V curve in anterior infarct and inversely a leftward shift in posterior infarction. The 115 patients of Group B are roughly equally distributed in the areas of normal, increased or decreased compliance. Any correlation was found between the extent of asynergy and changes in modulus of chamber stiffness. However, ejection fraction or VCF were higher in patients with a reduction of compliance than in patients with an increase of compliance.


Observations on the effect of angina on the left ventricle, with special reference to diastolic behavior. A series of investigations on patients developing angina following pacing are described. Evidence suggesting tension prolongation in the ischaemic areas of the ventricle with reduction in segmental systolic inward movement and a decrease in segmental diastolic complicance is presented and discussed.


Regional abnormalities of left ventricular wall movement during isovolumic relaxation in patients with ischemic heart disease. In order to study events during isovolumic relaxation, left ventricular angiograms of 120 patients with ischemic heart disease were digitized frame by frame, and compared with those of 15 normal subjects. In patients with ischemic heart disease, abnormal inward movement of endocardium occurred in areas supplied by narrowed coronary arteries. When these involved the free wall, they were due to abnormal wall thickening rather than to inward movement of epicardium. Since the volume of the ventricle was constant, they were accompanied by compensatory outward movement of endocardium elsewhere, due to premature thinning. Identical abnormalities were demonstrated in 80 patients by M-mode echocardiography, and in individual patients, agreement with angiography was good. These abnormalities were aggravated by TNT administration, and were unaffected by isometric stress. In approximately half, they were associated with abnormalities of isovolumic contraction. They appear to represent the behavior of regions of the left ventricle with partial loss of function due to previous ischemic injury.


Calcium transients and relaxation in single muscle fibers. Muscle contraction is initiated by an elevation in intracellular calcium. The transient change in free calcium to a brief depolarization, the calcium transient, can be recorded using a calcium luminescent protein, aequorin. The calcium transient precedes force, peaking while force is rising and returning to the resting level as peak force is achieved. In single barnacle muscle fibers microinjected with aequorin, shortening the muscle during the declining phase of the calcium transient produces an addition light signal, indicating extra free calcium in the sarcoplasm. The amount of additional light is larger with larger length changes. It is also larger if the shortening occurs early in the calcium transient rather than later. The amount of this extra calcium correlates well with the instantaneous level of the calcium transient and not with the instantaneous force level. It is argued in a speculative manner that this extra calcium is coming from the myofilaments. This supports the hypothesis that calcium binding to the myofilaments is rapid and reversible, that reaccumulation of calcium into the sarcoplasmic reticulum (SR) could occur long before relaxation begins and that relaxation of tension could occur by some process other than the mere removal of calcium from the myofilaments.


Identification of ionic currents underlying the repolarization process in the frog auricle. Voltage clamp experiments were performed to analyse the voltage and time dependence of the secondary inward current in frog atrial preparations. The fibers were treated with tetrodotoxin and tetraethylammonium or cesium to suppress the fast inward current and reduce the outward current; Cl- was replaced by CH3SO-4 to reduce the background current. Evidence is obtained suggesting that the inactivation process of the calcium dependent inward current is relatively rapid in the frog atrium. The current-voltage relationship of this current system appears to be nonlinear. The calcium dependent inward current transient is followed by a subsidiary net inward current which is presumably mainly Na+ dependent and underlies the depolarization tail of the action potential.


The inotropic action of adrenaline on cardiac muscle: does it relax or potentiate tension? Adrenaline has been shown to increase twitch tension and enhance relaxation in cardiac muscle. In mammalian myocardium, a unitary mechanism, namely facilitated uptake of calcium by the sarcoplasmic reticulum, is proposed to increase the internal recirculating store of calcium (thereby potentiating twitch tension) and simultaneously enhance relaxation. In frog ventricular myocardium, where tension is directly controlled by membrane potential, adrenaline seems to produce its positive inotropic effect by increasing the duration and amplitude of cardiac action potential plateau. If adrenaline is prevented from changing the action potential, either by electrical or pharmacologic means, the relaxant effect of the drug is unmasked. The results suggest that in frog ventricle, unlike mammalian myocardium, adrenaline may not have a 'true' positive inotropic effect independent of membrane potential. The findings in frog ventricle, where there is little or no internal recirculation of calcium, are consistent with the  model proposed for the mammalian myocardium.


Sarcomere relaxation in intact cardiac muscle. The process of tension decay in the absence of sarcomere motion was studied in intact heart muscle isolated from rats. Sarcomere length, observed by an infrared light diffraction technique, was controlled and the effect on force dissipation measured. Distribution of the diffracted light suggested that sarcomere length was uniform immediately after contraction than prior to the next. When sarcomeres - as distinct from the total muscle length - are kept isometric, the rate of tension decay (a) reaches its maximum sooner, and (b) appears to be constant throughout relaxation. Slow stretch (about 3% sarcomere length) prolonged cardiac muscle tension during early relaxation without direct evidence of sarcomere 'yielding'. The dynamics of isometric relaxation and the effect of stretch were qualitatively the same at external calcium concentrations of 0.6 and 1.9 mM. For a specified sequence of sarcomere length changes during relaxation, the dynamics of tension decay were independent of preloaded sarcomere length when the sarcomeres are kept isometric. The data suggest that lowering contractile tension enhanced relative relaxation rate (sec-1) at a sarcomere length of 2.0 micron. The results clarify the physiological mechanism regulating the dynamics of myocardial fiber relaxation.


Antihypertensive effect of various doses of hydrochlorothiazide and its relation to the plasma level of the drug. Nine previously untreated hypertensive patients (WHO Stage I and II) were given hydrochlorothiazide (hct) 12.5, 25, 50  and 75 mg for two weeks after an initial four week period of placebo treatment. Blood pressure recordings were made casually and after 30 min rest in lying, sitting and standing positions. Plasma concentrations of the drug were measured by GLC. A significant decrease in BP was seen during treatment with hct 12.5 mg except in the casual standing position. Doubling of the dose twice produced very little further decrease in BP. The mean steady state concentration was 111 ng.ml-1 during treatment with hct 75 mg. A linear relationship was found between the plasma concentration of hct at 0 and 5 h and the various dose level of hct. No relation was found between plasma concentration and reduction in blood pressure.


Kinetics and efficacy of theophylline in the treatment of apnea in the premature newborn. Aminophylline (theophylline-ethylenediamine) was administered to 27 premature newborns to prevent apneic spells. Of the 22 patients monitored for theophylline concentration, a therapeutic blood level was reached in 19 in 1--2 days, and 3 stayed below it. "Toxic" blood levels (less than or equal to 20 microgram/ml) were reached in 3 cases, one of whom showed signs of toxicity. Theophylline treatment was not efficient in the prevention of apnea when a serious underlying disease was present. Theophylline blood half-life (mean:27.0 h) and clearance (mean 12.9 ml/h/kg) confirmed the slow elimination pattern of the drug in the premature infant.


Paracetamol (acetaminophen) kinetics in patients with Gilber's syndrome. The pahrmacokinetics of paracetamol after intravenous and oral administration has been studied in 6 patients with Gilbert's syndrome, and 6 healthy controls. Paracetamol clearance was significantly less in the patients (255 ml/min SE +/- 23 ml/min) than in the normal subjects (352 ml/min SE +/- 40 ml/min). Moreover, whilst paracetamol concentrations declined monoexponentially in the patients, the decline was biexponential in the controls. No difference in the bioavailability of 500 mg paracetamol given orally was observed between the two groups. The results suggest that not only is paracetamol elimination impaired in Gilbert's syndrome, but that its distribution kinetics are also abnormal. Both these findings could be attributed to a decrease in hepatic glucuronyl transferase activity.


Reduction of absorption of digoxin, phenytoin and aspirin by activated charcoal in man. The inhibitory effect of activated charcoal 50 g suspended in water on the absorption of digoxin, phenytoin and aspirin was studied in six healthy volunteers in a cross-over manner. The absorption of digoxin and phenytoin were almost completely prevented (about 98%) when activated charcoal was ingested immediately after the drug. The total absorption of aspirin was inhibited by 70%; with clear postponement of absorption and partial release of aspirin from the charcoal in the gut: The peak serum concentration of aspirin was reduced by 95% by charcoal. When activated charcoal was ingested 1 hour after the drugs the inhibition of absorption was considerably less. However, since the absorption of larger doses of the drugs is often slow, the administration of an adequate dose of activated charcoal will be of definite value in the treatment of acute intoxication, even if delayed for several hours.


Preliminiary data on the pharmacokinetics of glaziovine in man. The pharmacokinetic parameters of Glaziovine, a pro-aporphine alkaloid with neuropharmacological properties, were investigated in healthy human volunteers. Glaziovine-14C 20 mg was administered in capsules (oral route) and in vials (i.v. route). Total radioactivity was meausred in plasma, urine and faeces. When administered orally, peak plasma levels were encountered at 2 h. The cumulative urinary excretion of total radioactivity over a 24 h period was 38% after oral and 50% after i.v. administration. Investigation of metabolites in urine revealed Glaziovine glucuronide as the sole metabolite of the drug. By comparing the percentage of urinary excretion or the area under the plasma level-time curve (AUC) obtained in the first 24 hours after i.v. and oral administration, enteral absorption was found to range from 78 to 84%. Thus, glaziovine appears to show very high enteratral absorption.


The placebo dilemma.

The placebo dilemma. Placebos have been used since about 1950 in evaluation and interpretation of drug efficacy, especially of new drugs, and in comparison with existing compounds to establish their clinical therapeutic value. There are a number of problems connected with use of an "inert nothing" the informed consent, which is required in a human experiment, the doctor's consciousness of deceitful behaviour, the potential risk for the patient and the doctor, the high effect rate of placebos (+/- 35%), unexpected side-effects, dependency of patients on placebos, discovery by the patient of the placebo treatment, factors independent of the medication, such as the doctor's bias, the nurses influence, the significance of colour, the inconsistency of placebo-reactor or non-reactor types and the genetic state of the patient, as well as the problems associated with a double blind test, which can hardly be used when a patient has previously received a medicine, such as morphine, with its euphoric effect, and the colour change produced in urine by drugs like rifampicin or riboflavin. Only a well-trained and experienced clinical observer should employ a placebo in establishing the therapeutic value of drugs whilst avoiding undesirable pitfalls.


Haemodynamic effects of molsidomin.

Haemodynamic effects of molsidomin. Hemodynamic changes for 30 min after intravenous administration of molsidomin 4 mg were studied in 10 patients. Pulmonary artery pressure, pulmonary wedge pressure, right atrial pressure and aortic pressure decreased significantly after 15 min, and after 30 min they were still below the control level. Heart rate increased significantly (by 15%) 15 min after injection, and after 30 min it was still 8% above the control value. Cardiac index decreased significantly from 3.37 +/- 0.12 to 2.95 +/- 0.15 1/min/m2. Peripheral and pulmonary resistance showed no significant change. Both stroke work index and minute work decreased significantly 15 min after administration of molsidomin and after 30 min both were still below the control level. The results suggest that molsidomin decreases preload without influencing afterload. By decreasing preload, molsidomin improves the oxygen supply/demand ratio of the myocardium.


Effect of the menstrual cycle, oral contraception and pregnancy on forearm blood flow, venous distensibility and clotting factors. Forearm blood flow, venous distensibility and various clotting factors were determined in 118 apparently healthy women (mean age 26 years), either during the menstrual cycle, or while taking a combined contraceptive (A) with high progestin:estrogen ratio (d-norgestrel 0.125 mg + ethinylestradiol 0.03 mg), or a sequential contraceptive (B) with low progestin: estrogen ratio (megestrol 0.1/1 mg + ethinylestradiol 0.1 mg), or in the 1st, 2nd and 3rd trimester of pregnancy. Venous distensibility in women taking contraceptive A was higher than in other women during the follicular phase of a normal menstrual cycle. Venous distensibility was not affected by contraceptive B. Blood flow and blood pressure remained unchanged by contraceptives A and B. Fibrinogen concentration was increased by both contraceptives, factor VII was either decreased (A) or unchanged (B), and factor X was either unchanged (A) or increased (B). The oral contraceptive with the high progestagen component appeared to increase venous capacitance and may induce venous stasis, whereas coagulability was particularly enhanced by the estrogen-type contraceptive. Pregnant women differed from women on oral contraceptives in regard to peripheral circulation; they showed a tremendous increase of blood flow with secondary vasodilation.


Effect of intravenous infusion of amitriptyline on total blood serotonin content. 18 female patients with primary depression received a slow intravenous infusion of amitriptyline and a good therapeutic response was observed in 15 patients as early as after 6 days. A highly significant correlation was found between improvement scores following the first and last infusions, which suggests that intravenous infusion of amitriptyline may serve as a prognostic test for prediction of drug efficacy. Total blood serotonin content prior to treatment was significantly lower than in normal controls. Amitriptyline infusion caused variable changes in blood serotonin and the changes were inversely correlated with pretreatment levels. A series of six infusions caused a gradual increase in baseline levels, together with a rapid decrease in the magnitude of the amitriptyline-induced changes in serotonin content.


Effect of sulphinpyrazone (Anturan) on uric acid excretion and plasma uric acid concentration in healthy volunteers. Investigations in six healthy volunteers given single oral doses of sulphinpyrazone (Anturan) showed that the drug had a potent uricosuric effect, even if the concentration of uric acid in the plasma was normal. In the dose range tested of 50 to 800 mg the increase in uric acid excretion was dose-related. It reached a maximum within 2 h and was still detectable 6--8 h after the drug was taken. There was a corresponding dose-related reduction in the plasma concentration of uric acid, which reached its lowest level after 8--10 h. The administration of 300 mg twice daily for four days to a further five healthy volunteers reduced plasma uric acid from 5.06 to 1.8 mg%. The findings indicate that, even if plasma uric acid levels are unknown, it would be advisable to avoid the risk of formation of urinary calculi by administering sulphinpyrazone in gradually increasing doses, ensuring adequate fluid intake and alkalinizing the urine during the first few days of treatment.


Metabolism of methaqualone in geriatric patients. The urinary excretion of five C-monohydroxy metabolites and the N-oxide of methaqualone has been measured in a group of eleven geriatric patients aged 71--90 years. The total excretion of the six metabolites in 24 h after the oral administration of a single dose was approximately one-half of that in a group of young healthy adults. The relative importance of the six metabolites was 4'-hydroxy greater than N-oxide greater than 2'-hydroxymethyl = 3'-hydroxy greater than 6-hydroxy = 2-hydroxymethyl which was the same order as that in young adults. The ratio of C-to N-oxidation was also the same in the two groups. There was no impairment of conjugation of the C-hydroxy metabolites with glucuronic acid in the geriatric group but there was greater interindividual variation in metabolite excretion. There was also evidence for delayed metabolism in the geriatric patients.


Inter- and intra-individual variation in the metabolism of methaqualone in man after a single oral dose. The urinary excretion of five C-monohydroxy metabolites and the N-oxide metabolite of methaqualone in the 24 h period immediately after oral dosing with 250 mg methaqualone (Melsed) has been measured in ninteen healthy adults (13 male, 6 female) to assess interindividual variations and in five adults (3 male, 2 female) on five separate occasions to assess intraindividual variation. The overall importance of the six metabolites was 4'-hydroxy greater than N-oxide greater than 2'-hydroxymethyl greater than 3'-hydroxy greater than 6-hydroxy = 2-hydroxymethyl. Variations in this order both within the 24 h period and within each of the three eight-hour periods constituting the 24 hours were minor and variations in the absolute amount of each metabolie excreted ranged from two to three-fold. Intraindividual variations were generally smaller than interindividual variations and for each individual the pattern of metabolism was similar on the five occasions. There is evidence that the C-oxidation of methaqualone may be more sensitive to cyclical variations in hormone levels than is N-oxidation.


m-Trifluoromethyl-alpha-ethylbenzhydrol: a new enzyme inducer. The enzyme inducing effect of m-trifluoromethyl-alpha-ethylbenzhydrol (RGH-3332) has been studied in man, rat and mouse. In man, RGH-3332 decreased the half-life of antipyrine and tolbutamide and increased D-glucaric acid excretion. In rat and mouse the half-life of antipyrine was shortened. It was concluded that in all three species man, rat and mouse the liver microsomal drug metabolizing enzyme system was induced by RGH-3332. No side-effects were found on treatment with RGH-3332.


Pharmacokinetic approach to intravenous procainamide therapy. A pharmacokinetic approach was employed to design a dosing regimen for the i.v. use of procainamide (PA) which consisted of a loading infusion given over one hour followed by a maintenance infusion. Therapeutic serum concentrations of PA were achieved in less than 15 min, and toxic serum concentrations were avoided in 12 patients. A mean maximum serum concentration of PA of 5.78 mg/l was obtained with a loading infusion of 16.6 mg/min PA HCl. An average steady-state serum concentration of PA of 5.05 mg/l was obtained with a mean maintenance infusion of 222 mg/hour PA HCl. The total body clearance of PA in slow and fast acetylators averaged 31 and 43 l/h respectively. Use of PA in cardiac patients by i. v. infusion can be safe and effective therapy.


Protein binding of salicylate in cutaneous hepatic porphyria. (1) Plasma protein binding of salicylate was studied in 14 patients with cutaneous hepatic porphyria (CHP) and 11 normal subjects using ultrafiltration with centrifugation (membrane cones) and continuous ultrafiltrations. (2) Albumin and haemoglobin levels were significantly reduced in patients with CHP, and salicylate binding by ultrafiltration/centrifugation was 65% compared with 84% in normal subjects. (3) Plasma porphyrin levels were raised, but did not correlate with salicylate binding, and protoporphyrin or uroporphyrin added to plasma did not alter the amount of drug bound. (4) Palmitate added to plasma reduced salicylate binding by 9 to 20% but a crossover of patient and normal plasma proteins and ultrafiltrates confirmed that no other ultrafiltrable metabolites present in patient plasma appeared to cause decreased binding. (5) Scatchard plots obtained by continuous ultrafiltration for normal and patient plasma showed a reduction in the number of primary and secondary binding sites and an increase in the intrinsic association constants for both these sites. (6) It was concluded that the decreased salicylate binding in CHP was due to a reduced albumin concentration and altered salicylate albumin interaction.


Effect of diazoxide on capacitance vessels. Haemodynamic changes after an i.v. bolus of diazoxide 300 mg were studied in 11 hypertensive subjects. A hypotensive effect due to a fall in total peripheral vascular resistance was found in all of them, but changes in the peripheral circulation were less regular. A uniform change in forearm blood volume, blood flow, vascular resistance and venous distensibility was not found after diazoxide. This is interpreted as being due to a reflex increase in sympathetic activity, which counteracts the direct vasodilator action of the drug.


Oral antipyretic therapy: evaluation of the N-aryl-anthranilic acid derivatives mefenamic acid, tolfenamic acid and flufenamic acid. The antipyretic activity of three N-aryl-anthranilic acid derivatives, mefenamic acid, tolfenamic acid and flufenamic acid, was compared and their optimal antipyretic dose determined in a trial in 87 children (aged 5 months to 15 years), who suffered from infections and fever exceeding 38.5 degrees C. Tolfenamic acid proved to be the most potent antipyretic agent of the three drugs; it was eight times more powerful than mefenamic acid and three times more powerful than flufenamic acid. The optimal antipyretic doses were: mefenamic acid 4 mg/kg, tolfenamic acid 0.5 mg/kg and flufenamic acid 1.5 mg/kg. It is evident that the antipyretic activity of these anthranilic acid derivatives is even greater than their antirheumatic effect, the difference being most noticeable in the case of tolfenamic acid.


Effects of various cephalosporins on the proximal tubule of the human kidney. Cephamandol 6.0 g, cephazolin 6.0 g or cephacetrile or cephalothin 8.0 g were administered as short-term infusions on 3 consecutive days to informed volunteers, who had no history or evidence of impairment of renal function. There were 15 subjects in the cephamandol, cephacetrile and cephalothin groups and 14 subjects in the cephazolin group. Alanine-aminopeptidase, a characteristic tubule enzyme, was determined in a 24-hour urine 2 days before administration, during the 3 day administration and on the 4 subsequent days. In addition, alanine-aminopeptidase was also estimated immunologically in concentrated urine with the aid of an anti-brush border antibody. Cephamandol, cephazolin and cephalothin were completely without effect on the proximal tubule. Cephacetrile, on the other hand, showed clear reactions in 9 out of 15 subjects, in the form of elevated AAP activity in urine and in 6 of the cases membrane elimination was demonstrable immunologically. After withdrawal of the medication, the values of the responder group returned spontaneously to normal, i.e. no cumulative effect was detected. These investigations show that elimination of alanine-aminopeptidase in the urine is a very sensitive index of the action of cephalosporins on renal tubules.


Pharmacokinetics and metabolism of the anilide local anaesthetics in neonates. 11 Etidocaine. The urinary elimination of etidocaine and several of its metabolites was investigated in neonates whose mothers had received one or more doses of etidocaine during labour. The urine collection period ranged among the neonates from 21.4 to 47.0 h post-partum. The total amounts of etidocaine and its metabolites recovered in neonatal urine represented a mean of 0.12% of the maternal dose. Some differences in the pattern of urinary metabolites were observed between neonates and adults. Mean half-life of elimination of etidocaine calculated from sigma-minus plots of the neonatal urinary data was 6.42 h. This is greater than that previously reported following intravenous administration of etidocaine to adults (2.6 h). The slower rate of elimination in neonates is probably due to an increased neonatal volume of distribution since there is evidence to show that etidocaine is extensively metabolised by the neonate.


Apparent volumes of distribution and drug binding to plasma proteins and tissues. A pharmacokinetic model that incorporates linear binding of drug to plasma proteins and tissue indicates the same relationship between apparent volume of distribution and drug binding as that proposed by Gillette (1971) based on a simple distribution model. Apparent volume of distribution (V) is directly proportional to free fraction of drug in plasma (fp) and indirectly proportional to free fraction of drug in tissue (fT). In the case of a constant fT, a plot of V versus fp will be linear with an intercept equal to plasma volume (Vp). If fT changes with fp, an apparently linear plot may result but the intercept will exceed Vp. An approach to the calculation of fT, a composite binding parameter, is presented and illustrated by comparing the tissue binding of tolbutamide in patients during acute viral hepatitis and upon recovery.


Influence of bed rest on the pharmacokinetics of phenazone. The pharmacokinetics of a single dose of phenazone was studied in six objects while ambulant and during bed rest for 3 days. Elimination of the drug was followed for 12 h after oral and intravenous administration. The elimination rate constant and total body clearance were significantly increased during bed rest as compared to the ambulant period, but the differences were small. The apparent volume of distribution decreased significantly. No consistent change due to bed rest was found in the rate of absorption or bioavailability of the oral dose.


Enhancement of the gastrointestinal absorption of hydrochlorothiazide by propantheline. Hydrochlorothiazide 75 mg was given twice p.o. to fasting subjects. In the second study they had been pretreated with propantheline 60 mg. Plasma and urine concentrations of hydrochlorothiazide were determined by GLC. Pretreatment with propantheline on average delayed the maximal plasma level of hct from 2.4 to 4.8 h (p less than 0.05); and the total urinary recovery of hydrochlorothiazide by 48 h was increased from 49.3 mg to 66.9 mg (p less than 0.005). It was concluded that propantheline increased substantially the absorption of the diuretic.


Measurement of plasma glycoside level following pengitoxin administration. 16-Acetyl-gitoxin, the cardioactive metabolite of pengitoxin, was estimated by the 86Rb uptake technique. In 70 consecutive patients taking an oral maintenance dose of pengitoxin 0.4 mg (Pentagit), the mean plasma concentration was 20.8 +/- 6.7 ng/ml. Toxic signs were not observed up to plasma levels of 40 ng per ml.


Immunity to transplantable carcinogen-induced fibrosarcomas in B2/B2 chickens. III. Tumor growth inhibition by local delayed hypersensitivity reactions to unrelated antigens. Delayed hypersensitivity (DH) reaction to human Ig, corynebacterium parvum or allogeneic cells at the site of tumor cell injection, suppressed the fibrosarcoma (SCFS) growth in chickens. Spleen cells of SC chickens sensitized to human Ig or C. parvum suppressed SCFS growth when adoptively transferred with tumor cells, but only when the sensitizing antigen was present locally. This suppression did not occur in irradiated recipients, SCFS I cells injected into wattles of chickens immune to the tumor, provoked a local DH  reaction. Spleen cells from donors sensitized to SCFS I adoptively transferred immunity to both SCFS I and SCFS II when cells of the two tumors were mixed together before injection, but not when SCFS II cells were injected alone. Tumor-specific and nontumor-specific DH may be essential for local suppression of fibrosarcoma growth in chickens.


Evidence for independent genetic regulation of the expression of different antibody classes in anti-sheep red blood cell responses. Three levels of variation are described in the response to sheep red blood cells (SRBC). Inbred strains of mice are distinguishable in terms of early or late kinetics of IgM response, high or low overall IgG response, and the relative expression of IgG1, IgG2a and IgG2b antibody in their response to SRBC. Results using C57BL/10 hybrid progeny strongly suggest a genetic control of these different aspects of the anti-SRBC response. The IgM kinetic pattern and the quantitative IgG response are regulated by two independent multigenic systems. Evidence is also presented for a distinction between the genes controlling the quantitative IgG response and those which control the 7 S isotypic pattern. IgG2a antibody expression seems regulated by a single gene. Neither group of genes involved in these various types of regulation seems directly linked to the H-2 complex or to the CH allotype. The hypothesis that 4 different sets of genes might control IgM, IgG1, IgG2a and IgG2b expression is discussed.


Antibody diversity in amphibians: inheritance of isoelectric focusing antibody patterns in isogenic frogs. Anti-sheep red cell, anti-dinitrophenyl, anti-phosphorylcholine antibody responses have been followed in isogenic frogs of the genus Xenopus. Isoelectric focusing antibody patterns show a high degree of overlap for all antigens studied, and a heterogeneity that is lower than in mammals for the same antigens. Inheritance of antibody isoelectric focusing spectrotypes was demonstrated for sheep red cells and dinitrophenyl in two clones of isogenic animals. Outbred frogs show a higher frequency of spectrotype sharing than outbred mammals. It is therefore suggested that antibody diversity is lower in frogs than in mammals.


The depolarizing action of 5-HT on mammalian non-myelinated nerve fibres. The effects of 5-hydroxytryptamine (5-HT) on nerve fibres in the rabbit cervical vagus and on the sciatic nerve "in vitro" were studied by the single sucrose-gap technique. The addition of 5-HT to the Locke solution bathing the vagus nerve induced rapid depolarization and a fall in spike height at the threshold concentration of 1 X 10(-7) M. In most cases the depolarizing effect was completely reversed by washing 5--10 min while in about 40% of the preparations the action potential amplitude remained 10--30% below the control level. These effects were dose-related up to a maximum concentration of 3 X 10(-5) M. Tachyphylaxis was not observed when the drug was added at 12--15 min intervals. Depolarization was abolished by perfusing the nerve with sodium-free medium or by previous exposure to lidocaine (10(-3) M). External hyperpolarizing currents (10(-7) to 10(-6) A) were not able to restore the action potential amplitude. Cyproheptadine (50 micron), which was found to have a slight local anesthetic action, reduced the 5-HT-induced depolarization by 20--30%. Methysergide (50 micron), a more specific 5-HT antagonist, did not affect the action of 5-HT. 5-HT was inactive when applied to the myelinated fibres of the sciatic nerve. Our results indicate that 5-HT-induced depolarization appears to be related to an increase in the resting sodium permeability of nerve fibres.


Methadone inhibits serotonin and norephinephrine uptake into rat brain synaptosomes and synaptic vesicles in vitro but not in vivo. Methadone in vitro inhibited biogenic amine uptake into synaptosomes isolated from rat whole brain; the IC50 for inhibition of serotonin uptake was between 0.1 and 1 micron, and for norepinephrine uptake, 10 micron when the serotonin or norepinephrine concentration was 0.05 micron. Methadone in vitro also inhibited uptake of norepinephrine into rat brain synaptic vesicles to the same extent that it inhibited synaptosomal norepinephrine uptake. In all cases, all cases, morphine in vitro was much less effective in blocking uptake. However, in vivo acute or chronic administration of methadone to rats failed to cause inhibition of serotonin or norepinephrine uptakes in synaptosomes isolated from the brains of treated rats, nor was inhibition of vesicular norepinephrine uptake produced. Amine uptake in situ also was unaffected by methadone administration, as evidenced by normal incorporation of intracisternally administered 3H-serotonin into synaptic endings. These data show that inhibition of synaptosomal and vesicular biogenic amine uptake by methadone in vitro is probably not a major determinant of the alterations in biogenic amine turnover caused by methadone administration. Thus, results obtained with platelets and adrenomedullary chromaffin vesicles, in which methadone administration does cause effective inhibition of amine uptake in vivo, cannot be extended to the central nervous system.


Effects of phenytoin on sympathetic reflex responses in the cat. Phenytoin (DPH), in a low dose (2 mg/kg) was found to potentiate reflex elevations in blood pressure and responses of the nictitating membrane induced either by the electrical stimulation of the sciatic nerve or by intravenously applied pentetrazol in light anaesthetized cats. Pressor responses to carotid occlusion under chloralose anaesthesia were also potentiated. In contrast a high dose (20 mg/kg) of DPH exerted an inhibitory effect on these responses. These effects of DPH on sympathetic reflex responses proved to be long-lasting. Intravenously applied picrotoxin (0.05 mg/kg) or the same dose of ouabain, injected into the vertebral artery, partially reversed the inhibitory effect of DPH (20 mg/kg). Our data support the suggestion that DPH exerts its inhibitory action by effecting ionic fluxes in the CNS, mostly in inhibitory structures.


Indomethacin and diclofenac sodium increase sodium and water excretion after extracellular volume expansion in the rabbit. The renal effects of an acute extracellular fluid volume expansion (50 ml Ringer/kg body weight/60 min) were studied in aldosterone-treated (100 microgram/kg), anesthetized rabbits with and without pretreatment with either indomethacin (3.0 mg/kg) or diclofenac sodium (3.0 mg/kg), two different inhibitors of renal prostaglandin (PG) biosynthesis. In controls (n = 7), the volume expansion increased urine flow from 1.5 +/- 0.24 to 6.1 +/- 0.5 (S.E.) ml/min/100 g kidney weight and sodium excretion from 0.15 +/- 0.03 to 0.99 +/- 0.10 mmol/min/100 g. PAH and inulin clearance increased by 42 and 58%, respectively, while plasma renin activity and urinary excretion of PGF2 alpha-like immunoreactivity were reduced (P less than 0.05). In animals pretreated with indomethacin (n = 6) or diclofenac sodium (n = 6), the diuresis and the natriuresis following volume expansion were significantly increased about two-fold over controls, whereas PAH and inulin clearance, plasma renin activity and hematocrit did not differ from controls. Both drugs were found to reduce urinary excretion of PGF2 alpha-like immunoreactivity by 75--95% througout the experiment. The results indicate that diclofenac sodium, indomethacin and extracellular volume expansion enhance sodium and water excretion partly by suppression of a PG sensitive reabsorption process in the kidney.


Some effects of disopyramide and its N-dealkylated metabolite on isolated nerve and cardiac muscle. In animals and man the antidysrhythmic agent disopyramide in primarily metabolised by mono-N-dealkylation. The effects of disopyramide and its N-dealkylated metabolite (MIP) have been investigated using isolated cardiac and nervous tissue, and their effects have been compared with the effects of other antidysrhythmic agents. Disopyramide, d,l-propranolol and quinidine all decreased both maximum driving frequency and developed tension in electrically driven guinea pit atria. MIP and procaine amide also decreased maximum driving frequency, but had a positive intropic effect. MIP was only 4 times less active than disopyramide in decreasing maximum driving frequency. There was no evidence that either disopyramide or MIP possessed beta-adrenoceptor antagonist properties. In superfused rat sciatic nerves, it has been shown that neither disopyramide nor MIP possesses significant local anaesthetic properties. Procaine amide and lignocaine were highly active in this test. The possible contribution of MIP to the actions of disopyramide in vivo is discussed.


Comparison of the thermoregulatory responses to intracerebroventricularly injected dopamine, noradrenaline and 5-hydroxytryptamine in the goat. The thermoregulatory responses to dopamine (DA), noradrenaline (NA) and 5-hydroxytryptamine (5-HT), injected into the third cerebral ventricle of goats at 20 degrees C ambient temperature (Ta) or during cold exposure, were compared before and after pretreatment with the DA receptor blocker haloperidol or the 5-HT receptor blocker methysergide. At 20 degrees C Ta, intracerebroventricular (i.c.v.) injection of DA (800 microgram), NA (200 microgram) or 5-HT (800 microgram) induced a decrease in body temperature (Tb) and dilatation of the ear vessels. After DA and 5-HT, but not after NA, panting was observed. During cold exposure both DA and NA caused a suppression of shivering and a fall in Tb. Pretreatment with haloperidol (400 microgram, i.c.v.) attenuated the thermoregulatory effects of i.c.v. DA other than the dilatation of the ear vessels at 20 degrees C Ta. Haloperidol did not influence the responses after i.c.v. NA or 5-HT. Methysergide (1.0 mg, i.c.v.) blocked panting and attenuated the decrease in Tb caused by i.c.v. DA and 5-HT at 20 degrees C Ta and blocked the peripheral vasodilatation caused by 5-HT but not the dilatation caused by DA. During cold exposure methysergide antagonized the thermoregulatory effects of DA, but not those of NA. Ic.v. injection of haloperidol (400 microgram) or methysergide (1.0 mg) during heat exposure induced in panting and a rise in body temperature, which suggests that both DA and 5-HT have a physiological role in the mediation of heat loss in the goat. We conclude that in the central thermoregulatory system of the goat are excitatory DA receptors in the pathway from heat sensors to heat effectors. Activation of these receptors by i.c.v. DA or by heat exposure results in a secondaary release of 5-HT. Since the vasodilating effect of DA was not influenced by haloperidol or methysergide, this effect could be mediated by haloperidol-insensitive (possibly inhibitory) DA receptors on the pathway controlling peripheral vasomotor tone. The thermoregulatory effects of i.c.v. NA are probably mediated by inhibitory NA receptors on the pathway from cold sensors to heat production effectors.


Successful oxime therapy one hour after soman intoxication in the rat. The bisquaternary mono-oxime HI-6, and to a lesser extent HS-6, caused functional recovery of neuromuscular transmission in vivo and in vitro when given 60 min after soman, i.e. when the soman-cholinesterase (AChE) complex is said to be fully "aged". Atropinised rats, with the tracheas intubated, received 4 X LD50 soman i.v. and were kept alive by artificial respiration. 60 min later HI-6 was administered and after an additional 15 min the tracheal tube was removed. Nearly all animals survived for 24 h. After 6 X LD50 soman followed by HI-6, HI-6, respiratory failure was delayed for hours but almost all animals died within 24 h. Against equal doses of soman, HS-6 was less effective. In experiments with isolated rat phrenic nerve-diaphragm preparations, HI-6 given 60 min after soman produced functional recovery which could be abolished by a second dose of soman, suggesting that HI-6 had reactivated the AChE and that this enzyme was then reinhibited by the second dose of soman. HI-6 reactivates purified bovine erythrocyte AChE when added immediately after inhibition by soman, but does not reactivate tabun-inhibited AChE. Accordingly, no functional recovery of neuromuscular transmission was found in rat diaphragm when HI-6 was administered 60 min after tabun. Furthermore, functional recovery was not obtained with HI-6 after exposure diaphragms to S 27, which carries a hydroxyl group instead of an alkoxy group--i.e. it is "pre-aged"--and instantaneously forms an inhibitor--enzyme complex identical to the "age" soman--enzyme complex. These results exclude the possibility that the functional recovery was caused by a direct pharmacological effect of the oxime. The functional recovery of diaphragms treated with II-6 60 min after exposure to soman was not accompanied by a return of histochemically detectable AChE activity. The capacity of the isolated diaphragm to hydrolyze (3H)-acetylcholine, however, seemed to be reactivated to a very small (1--2%) extent by HI-6, 60 min after exposure to soman. It is concluded that soman-inhibited cholinesterase in intact rat tissue "ages" much more slowly than does soman-inhibited purified cholinesterase, so that even after 60 min enough "non-aged" inhibited AChE is still susceptible to reactivation to be lifesaving.


Studies on mast cells of Rana tigrina.

Studies on mast cells of Rana tigrina. Mast cell degranulators cause vasoconstriction in the frog. A comparative study of the mesenteric mast cells of the frog and rat showed that the frog mast cells differed from those of the rat in having a dense perigastric instead of a sparse ideal distribution, and in responding to compound 48/80 by a rapid and marked disappearance and/or fragmented appearance instead of classical degranulation. The vasoconstriction and mast cell response after in vivo compound 48/80 appeared related since they showed a parallel increase when the dose was increased.


Disappearance of cortical granules in rat ovum in vivo following fertilization. Electron microscopical studies of the rat ova were carried out to clarify the pattern of disappearance of cortical granules following fertilization. When the posterior cap of the head of a spermatozoon was attahced to the vitelline membrane, cortical granules located beneath this membrane fused with this membrane to be decomposed or broken. Then their contents were discharged into the perivitelline space. The disappearance of cortical granules seemed to have started in an area around the site of the vitelline membrane to which spermatozoon was attached and spread soon all over the vitellus.


[Comparative studies on lactate dehydrogenase in serum and plasma of rats (author's transl)]. Values of serum and plasma LDH in rats were comparatively studied, and the following results were obtained: 1) The activity of LDH increased in serum with time during clotting, but no changes of LDH activity were found in plasma. 2) When platelet rich plasma (PRP) was recalcified and allowed to clot, LDH-release from platelets with a corresponding increase of serum LDH was observed, but addition of ADP or thrombin to PRP did not have an effect on LDH-release. 3) LDH-release from platelets by calcium was not inhibited by aspirin, and it was influenced by the quality of the test tube. 4) Values of serum and plasma LDH on experimentally induced liver-damaged or kidney-damaged rats and tumor-bearing rats were examined in relation to their tissue damages, revealing that plasma LDH activity represents the condition of a disease better than serum LDH activity.


[Three cases of hemolytic Streptococcus infection in guinea-pigs (author's transl)]. Three cases of naturally occurring hemolytic streptococcus infection were detected in guinea-pigs which had been delivered from different sources to the Division of Animal Research, Faculty of Medicine, University of Tokyo, Tokyo, during the period from November, 1976, till February, 1977. The clinical manifestations of the infection were differentiated into two types; ie, the acute type with sepsis and pneumonia as main pathologic changes, and the chronic type with abscess formation of submaxillary lymph node. Streptococcus zooepidemicus was isolated from the conjunctiva of about 56% of these guinea-pigs. Healthy guinea-pigs housed with spontaneously infected ones in the same cage suffered from the infection, showing manifestation and pathologic changes similar to the spontaneous cases. Some of them, however, remained apparently healthy for about 2 months harboring the organisms in the conjunctiva or nasal cavity. It is presumed that such carriers become a source of the infection in nature.


[The reliable diagnosis for an early stage of hereditary muscular dystrophy of mice (author's transl)]. The methods to find hereditary muscular dystrophy mice (dy-mice) were described. One was to observe whether or not mice dragged their hind limbs immediately after they were dropped onto a table following their suspension by their tails. The other was to observe dragging their hind  limbs immediately after they were restored to a normal position from a supine one. This limbs-dragging induced by these methods were characteristic of dy-mice. Among 437 mice checked by these methods, 76 showed the drag during 14-23 days after birth (18.3 days on the average) and later they were confirmed to be dy-mice without exception. Therefore, it was concluded that these methods were useful for the accurate diagnosis of dy-mice at an early stage.


The effect of Rauscher murine leukemia virus infection on the hemopoietic system of BALB/c mice. Cell proliferation and cell loss. Cell proliferation was investigated in normal and Rauscher Leukemia Virus-infected BALB/c mice. Five days after inoculation, islands of leukemic blasts arose in the red pulp, and proliferated as shown by autoradiographic analysis after a pulse of 3H-Thymidine. These cells subsequently infiltrated the whole spleen and 3 weeks after infection about 60% of the spleen consisted of large immature erythroblast-like cells. Repeated injections of 3H-Thymidine led to uniform labeling of 85% of the spleen cells. Cell cycle analysis showed that for bone marrow as well as for spleen cells the total duration of the cell cycle did not differ from the cell cycle times of normal erythroblasts. From the difference between the actual doubling time and the potential doubling time (estimated on the basis of the cell cycle time) it can be calculated that considerable cell loss must occur. This cell loss is only to a minor extent due to the release of blasts into the peripheral blood. Probably cell death and extrusion of nuclei during erythroid differentiation are the main factors involved.


Improved technique for increased granulocyte recovery from canine whole blood samples by counterflow centrifugation-elutriation. I. In vitro analysis. Granulocytes were isolated from 120 ml of canine whole blood by a modification in counterflow centrifugation-elutriation technique. The leukocytes were concentrated in a buffy coat fraction and diluted to a fixed RBC density prior to entry into the rotor. Overall recovery of granulocytes from canine whole blood averaged 82% with 96% purity from contaminating mononuclear leukocytes. The purified cellular fraction was assayed in vitro to monitor O2 consumption, chemotaxis, bacterial growth inhibition, and enzyme activities. Cellular integrity was monitored by scanning and transmission electron microscopy as well as by cell volume analysis. The data suggest that the granulocytes isolated by counterflow centrifugation-elutriation suffered no noticeable morphological damage or loss of function in terms of recognition of a toxic agent, migration, phagocytosis, or bactericidal capacity. Granulocytes isolated by counterflow centrifugation-elutriation may be physiologically more active than cells obtained by filtration leukapheresis, continuous-flow centrifugal leukapheresis or discontinuous flow centrifugation both in vitro and in vivo.