Effects of cis-platinum(II) diamminedichloride on survival and the rate of DNA synthesis in synchronously growing Chinese hamster V79-379A cells in the absence and presence of caffeine inhibited post-replication repair; evidence for an inducible repair mechanism. The mechanism by which cis-Platinum(II)diamminedichloride, (cis-Pt(II)), inhibited DNA synthesis was investigated using synchronously growing Chinese hamster V79-379A cells. The progression of G1 phase treated cells into S phase was not inhibited. However the rate of DNA synthesis throughout the S phase was depressed in a dose dependent manner compared to a control. The peak rate of DNA synthesis was delayed and the duration of the S phase was extended resulting in a dose dependent delay in the timing of mitosis in the first cell cycle after treatment. The delay in the timing of mitosis after the second S phase in treated cells was comparable to the delay observed after the first cycle. Post treatment incubation of cells in the presence of a non toxic concentration of caffeine resulted in an alteration in the pattern of DNA synthesis, leading to a reversal of the dose dependent delay in the timing of the peak rate of DNA synthesis to a position corresponding to the control. Caffeine potentiated the toxicity of cis-Pt(II) only if it was added during the first cell cycle after treatment. The patterns of DNA synthesis in synchronous cultures of Chinese hamster V79-379A cells after cis-Pt(II) treatment appeared to be similar to those previously demonstrated for difunctional alkylating agents. The results were discussed in relation to an existing model in which it was postulated that a DNA replication dependent repair mechanism enables cells to replicate DNA using a cis-Pt(II) damaged DNA template. This mechanism has been shown in Chinese hamster V79-379A cells to be sensitive to non toxic concentrations of caffeine resulting in an inhibition of the rate of ligation of nascent DNA which is followed by an increase in chromosome aberrations and cell death. The results presented support the proposed model and emphasise the importance of this replication repair mechanism in enabling cells to tolerate unexcised damage to their genome. In addition we propose that cis-Pt(II) damage stimulates the induction of a system which is involved in enhancing so called post replication repair in Chinese hamster V79 cells and that this induction might be inhibited by the presence of caffeine.


Characterization of aminoalkylphosphonyl cerebrosides in muscle tissue of Turbo cornutus. From muscle tissues of the marine snail (Turbo cornutus) aminoalkylphosphonyl cerebrosides, which had been shown to be present in visceral parts were isolated. Their structure was determined by degradative methods and by characterization of components by gas chromatography-mass spectrometry. The aminoalkylphosphonyl cerebroside fraction consisted of a major portion of 1-O-[6'-O-(N-methylaminoethylphosphonyl) galactosyl] ceramide and a minor portion of a novel lipid, 1-O-[6'-O-(aminoethylphosphonyl)galactosyl] ceramide. The fatty acids of the fraction were mainly palmitic (53.3%) and 2-hydroxy palmitic acid (14.6%). The long chain bases were mainly dihydroxy C22: 2(36.6%), C18:1 (14.6%) and trihydroxy bases were also found as minor components.


[Percutaneous transhepatic biliary drainage]. The authors describe their experience in 23 cases of biliary tract drainage by the transhepatic-percutaneous approach in the course of obstructive jaundice of diversified origin. This can be done for essentially three reasons, namely to alleviate jaundice preoperatively, to provide permanent bile drainage in patients not amenable to surgery, and to relieve excess pressure in surgical anastomoses of the biliary passages. On the basis of biological considerations (relationship between severity and duration of cholestasis on the one hand and postoperative mortality and morbidity on the other), and in light of their own results, the authors argue in favor of this procedure, explaining that it is only mildly traumatic to the patient, easy to perform, attended by a low quota of complications, and above all effective as a drainage; also, it does not unduly prolong the preoperative period for patients scheduled for further and major surgery. Also in view of the current role of PTC in the diagnosis of obstructive jaundice, they submit that transhepatic-percutaneous drainage should be done right next to recognition of dilatation of the intrahepatic bile passages by CAT or echotomography.


[Surgical treatment of caval obstructions]. The vena cava is the seat of obstructive disorders for which direct reparative surgery may sometimes become necessary. S.C.S. is mainly caused by neoplastic invasions or, in the case of the inferior vena cava, by progression of phlebothrombosis of the lower limbs. Prosthesis or autologous substitution and thrombointimectomy are specifically indicated in the two cases respectively. The two methods are described in detail and exemplified through the authors' personal cases consisting in an inferior caval thrombointimectomy and four superior caval by passes. It is concluded that surgical procedures are effective when the primitive disorder can be dominated and the thrombosis is not excessively extensive.


[Incidence of recurrent nerve paralysis in personal cases of thyroid surgery]. The authors review their series of thyroid operations, totaling 383 cases in the last seven years. In that series the incidence of recurrent nerve lesions leading to permanent sequels was 1.07% (all treated without surgical preparation of the nerve); an additional 0.53% of the patients developed transient paralysis of the recurrent nerve--of which the authors discuss the possible etiology. The authors conclude by stating the need for careful recognition of the recurrent nerve in all surgical procedures involving the thyroid gland, except some enucleo-resections; they confirm the importance of branches of the inferior thyroid artery and the inferior horn of the thyroid cartilage as reference points.


[Current views concerning the etiopathogenesis of reflux esophagitis in children]. Esophagitis is caused by a gastro-esophageal reflux sometimes reflecting anatomical inadequacy of structures that secure the relationships between the cardia and the diaphragm, fundus of the stomach, and other surrounding viscera, but more often due to functional deficiency of lower esophageal sphincter (LES), this representing the main factor for maintenance of the mechanism of cardial continence. Manometric studies of intraluminal pressures in the esophagus have revealed the presence of a high-pressure segment in the distal esophagus, where a mean pressure of 12-13 mm Hg obtains over a length of 3 or 4 cm. In the first two weeks of extrauterine life the LES is between 0.5 and 1 cm long, with a pressure of 3 mm Hg; pressure values in the LES equal adult values at about one month of age, showing that neuromuscular control of the sphincter has been achieved. Gastro-esophageal reflux, however, is very common in the newborn and not enough by itself to cause peptic disease of the esophagus; this requires the concomitance of morphological defects of the esophageal mucosa and/or dynamic-functional disturbances. This, according to the authors, is why esophagitis actually occurs in only about 60% of patients with radiologically and pHmetrically demonstrated gastro-esophageal reflux.


[Clinical picture and diagnosis of reflux esophagitis in children]. Whereas a burning retrosternal pain irradiating upward is the main symptom of reflux esophagitis in the adult, this symptom is usually not extant in children. Rather, the child will show unaccountable vomiting, stunted growth, anemia of unknown etiology, and respiratory disturbances. The pathologic condition more often associated with reflux esophagitis is hypertrophic stenosis of the pylorus, which is in fact regarded as one of the causes of the esophageal disorder. Reflux esophagitis may lead to ulceration and hemorrhage, and may evolve into cicatricial retraction and esophageal stenosis, the latter sometimes quite tight. The condition is diagnosed in light of radiological examination and endoscopy. Whit older children, one may add the Bernstein test and pressure and pH readings. The paper concludes by pointing out the different diagnostic approach in the newborn and small baby on the one hand and in older children on the other.


[Current trends in the treatment of reflux esophagitis in childhood]. A problem of some complexity is the management of pathological gastro-esophageal reflux due to the presence of cardio-hiatal malformation, malposition of the cardia and gastric tuberosity, a congenitally short esophagus, and the like. Sometimes, however, there is no demonstrable morphological aleration and the condition, characterized by primitive hypotonia of the LES, goes under the name of infant chalasia of the esophagus. The general policy is to try first a conservative treatment consisting of dietary and postural measures and the administration of metoclopramide and d;ugs that protect the esophageal mucosa; this stage, however, should not be prolonged beyond 6 weeks. The presence of gastro-esophageal reflux associated with hiatal hernia; the presence of severe esophagitis or peptic stenosis of the esophagus and the persistence of symptoms after an adequate period of conservative therapy constitute as many indications for surgical correction. Good results can be obtained by restoring or strengthening the failing function of the LES, as is done quite successfully with funduplication after Nissen or with method of Belsey-Mark IV.


[Personal experience with anterior resection of the rectum]. Authors experience in the Dixon's operation regarding the surgical treatment of cancer of the rectum and recto-sigmoid junction is reviewed. It consists in 224 cases of malignant lesions which were treated by Miles operation (106 cases--47%), Dixon's operation (49 cases--21%), palliation anterior resection (16 cases--7%), other operations (53 cases--24%). After a brief discussion on anatomo-pathological conditions and tecniques, the Authors consider their results: operatory mortality 4%; complications: total anastomosis dehiscence 1 case, partial anastomosis dehiscence 14%; local repetitions 10%; hepatic metastasis within 5 ys 40%; survival at 5 ys 46%. Functional results were always very good. The Authors conclude that, even considering the large rate (56%) of wide-spread cancers treated, Dixon's operation, at least in their experience, doesn't seem to satisfy those needs of radically requested by a correct surgical therapy, which only Miles' operation guarantees.


[Current criteria of treatment of angiomas]. The authors set out their therapeutic trend in the treatment of angioma and the criteria governing the use of the various therapies according to the type of dysplasia and its clinical course; they then report on the results obtained in a series of 400 cases. Only 22 cases with unfavourable trend (evolution of lesion) were observed, whereas the others exhibited spontaneous regression (85 cases), arrest of evolution (50 cases) or regression or cure (243 cases).


[Behavior of mast cells in experimental burns in rats]. The Authors, using a personal dyeing method, specific for mast cells, that they described in a former work, (a modification of May-Gruenwald-Giemsa dye) observed crowd of mast cells in the subcutaneous tissue, below the site of thermal skin burns, in rats. They clearly demonstrate the phenomenon of mast cells degranulation. The granules are released in the subcutaneous tissue, then they go away along the limph vessels, and it is possible to follow them until the limph nodes. There is also crowd of limphocytes and plasma cells; nevertheless, there is evidence that mast cells are the early cells which react to burns in rats. Perhaps it should be possible that mast cells have some immunological function, owing to the contact their granules take with lymphocytes and plasma cells.


[Cavernous hemangioma of the mediastinum]. The authors present a rare case of cavernous hemangioma of the mediastinum. After a critical review of existing literature on the subject, they discuss the symptoms of this vascular malformation with special emphasis on the difficulties of differential diagnosis versus other space-occupying lesions of the mediastinum. They further discuss the characteristic histopathologic features of cavernous hemangioma, and state that ablative surgery is the only possible therapeutic measure.


[Surgical treatment of rheumatoid arthritis]. The Authors presents a review of the orthopedic surgery in patient with rheumatoid arthritis. The aims of surgical treatment in this patients are: an improvement of the function, relief of pain and improvement in appearance. In this study are presented the major types of operation currently used: synoviectomy, arthroplasty, repair of tendons, osteotomy and arthrodesis.


[A fairly common complication of acute appendicitis (a propos of 42 cases of abscess of Douglas' pouch)]. After a brief review of the literature, 42 cases of Douglas-pouch peritonitis are described, with particular regard to those cases arosen as a complication of acute appendicitis. Symptomatology and anatomo-pathological pictures are carefully described and the most likely pathogenetic causes are discussed. Particular attention is paid to the surgical techniques used.


Role of the renin-angiotensin system in the systemic vasoconstriction of chronic congestive heart failure. In 15 patients with severe chronic left ventricular failure, plasma renin activity (PRA) ranged widely, from 0.2--39 ng/ml/hr. The level of PRA was unrelated to cardiac output (CO) or pulmonary artery wedge pressure (PWP), but was slightly negatively correlated with mean arterial pressure (MAP) (r = -0.45) and systemic vascular resistance (SVR) (r = -0.40). After infusion of the angiotensin converting enzyme inhibitor teprotide (SQ 20,881) PWP fell from 26.3 +/- 1.3 (SEM) to 20.3 +/- 1.4 mm Hg (P less than 0.001), CO rose from 3.94 +/- 0.23 to 4.75 +/- 0.31 l/min (P less than 0.001), MAP fell from 87.5 +/- 3.8 to 77.9 +/- 4.1 mm Hg (P less than 0.001) and SVR from 1619 +/- 148 to 1252 +/- 137 dyne-sec-cm-5 (P less than 0.001). The fall in MAP and in SVR was significantly correlated with control PRA (r = 0.68 and r = 0.58, respectively). When subjects were divided on the basis of control PRA the hemodynamic response to teprotide was greatest in the high renin group. PRA rose after teprotide (8.7 +/- 3.4 to 37.9 +/- 7.7 ng/ml/hr, P less than 0.05) but plasma norepinephrine fell (619.1 +/- 103.6 to 449.7 +/- 75.7, P less than 0.05). The renin-angiotensin system thus appears to have an important role in the elevated SVR in some patients with heart failure. Chronic inhibition of converting enzyme should be explored as a possible therapeutic approach.


Angiotensin converting enzyme inhibition in patients with congestive heart failure. The etiology of afterload elevation in congestive cardiac failure is unclear, but experimental evidence suggests a role for the renin-angiotensin system in maintaining elevated peripheral vascular resistance. The angiotensin converting enzyme inhibitor SQ20,881 was administered to eight patients with congestive cardiac failure (four hypertensives, four normotensives) during or one day after diagnostic cardiac catheterization. Various hemodynamic measurements performed before and during blockade indicate that this agent caused improvement in cardiac function in all patients by decreasing afterload. This improvement correlated with the decrease in total vascular resistance but was independent of the baseline blood pressure and plasma renin activity. These results suggest that inhibition of angiotensin converting enzyme is a worthwhile approach to the treatment of congestive heart failure, although its exact mechanism of action remains unclear.


Comparison of single-dose and double-dose thallium-201 myocardial perfusion scintigraphy for the detection of coronary artery disease and prior myocardial infarction. Thallium-201 myocardial perfusion scintigraphy was performed after exercise, 4 hours after exercise (redistribution) and after a separate rest injection in 87 patients undergoing coronary arteriography. Significant coronary lesions were present in 62 of the patients. Interpretation of the rest and redistribution scintiscans was the same in 69 patients, 45 of whom had coronary artery disease (CAD). In 16 of the 17 patients with CAD and differing interpretations, defects were present on redistribution scintiscans but not on rest scintiscans; 11 of these patients had evidence of prior transmural myocardial infarction and the other five had an occluded coronary artery supplying the region of the defect. Redistribution scintiscans were more sensitive than rest scintiscans for the detection of prior myocardial infarction (93% vs 54%; P less than 0.01). The increased sensitivity was confined to the detection of prior inferior myocardial infarctions. In 36 of 38 patients with persistent perfusion defects on 4-hour redistribution scintiscans, either a prior infarction or an occluded coronary vessel was present. Exercise scintiscans were compared with rest scintiscans or with redistribution scintiscans for the detection of CAD. The sensitivity was not significantly different with either technique (90% and 89%, respectively), but both scintigraphic techniques were more sensitive than exercise electrocardiography (66%, P less than 0.01). These data demonstrate that redistribution thallium-201 scintiscans may be substituted for conventional rest scintiscans, resulting in reduced cost and radiation exposure to the patient.


Assessment of cardiac performance with quantitative radionuclide angiocardiography. Effects of oral propranolol on global and regional left ventricular function in coronary artery disease. The effect of incremental dosages of oral propranolol (mean peak dose of 165 +/- 13 mg/day) of left ventricular ejection fraction, ejection rate and regional wall motion was studied sequentially in 22 stable, resting patients with coronary artery disease using a geometry-independent first-pass radionuclide angiocardiographic technique. All patients improved clinically, in association with a fall in heart rate and therapeutic serum propranolol levels. No significant changes were noted in ejection fraction, ejection rate or regional wall motion. No patient developed a new regional wall motion disturbance. Thus, oral propranolol administered at clinically effective antianginal dosages in patients with stable coronary artery disease does not appear to have significant deleterious effects on resting left ventricular performance.


Staged surgical management of tetralogy of Fallot in infants. A cohort of 61 consecutive patients 24 months of age of younger had palliative shunts for symptoms of tetralogy of Fallot during a 12-year period. Thirty-six of these patients have been followed through definitive intracardiac repair or to death. For analysis palliative operations were separated into two six-year periods, 1965--1970. During the first period seven of 30 infants operated on died; all 31 infants operated on during the second period survived. The Waterston anastomosis was performed most frequently (67%) during the first period; the Blalock-Taussig anastomosis was performed in 68% of infants during the second period. Of 54 hospital survivors, three died before definitive intracardiac repair. Two of the three interim deaths were related to heart disease. Twenty-six of the remaining 51 patients have had definitive intracardiac repair with two deaths (8%). Twenty-four in this group had intracardiac repair since 1973 with one hospital death (4%). The cumulative mortality for the entire cohort is 25%, but more recent experience (1971--77) indicates a cumulative mortality near 5%. The recent mortality rate for staged management is less than the 14% rate reported by others for primary intracardiac repair of tetralogy of Fallot in 205 infants. We conclude that primary intracardiac repair has important advantages for infants with tetralogy of Fallot who have favorable anatomic features and no other associated cardiac lesions or medical problems. Staged management of tetralogy of Fallot is still recommended for infants with unfavorable anatomy, additional lesions or associated medical problems.


Noninvasive assessment of left ventricular function from the mitral valve echogram. Relation of final anterior mitral leaflet closing velocity to peak dp/dt and aortic velocity. Since final mitral valve (MV) closure and aortic ejection velocity are mediated by the same forces in early left ventricular (LV) contraction, the rate of final MV closure (BC slope) should reflect LV performance. We first verified whether peak final closing velocity (ds/dt) of the anterior MV leaflet (AMVL) is related to peak aortic ejection velocity (V) and LV dp/dt in 18 open-chest dogs. We then checked the validity of the relations in man. Our approach was to measure peak ds/dt, peak aortic acceleration (dV/dt) and peak LV dp/dt using electronic differentiation of analog signals of the AMVL echogram, V and LV pressure. In dogs, resting ds/dt averaged 26.9 +/- 9.0 (SD) cm/sec and changed significantly (P less than 0.001) after isoproterenol, propranolol, coronary ligation and aortic cross-clamping. We found good (P less than 0.001) correlations between ds/dt and V (r = 0.82), dV/dt ( r = 0.67) and dp/dt (r = 0.73). In man, resting ds/dt averaged 25.5 +/- 1.6 cm/sec in six normals. In 40 patients with coronary artery disease, restind ds/dt was lower (15.7 +/- 4.4 cm/sec; P less than 0.001) in the 19 with resting LV end-diastolic pressure (LVEDP) greater than 12 mm Hg. Resting ds/dt correlated closely with V (r = 0.82, N = 10), dp/dt (r = 0.93, N = 6), resting LVEDP (r = -0.67, N = 40), angiographic ejection fractions (r = 0.62, N = 40) as well as manually obtained BC slopes (r = 0.93, N = 40). Thus, final MV closing velocity provides  a useful and simple means for the objective noninvasive assessment of LV performance.


Echocardiographic features of constrictive pericarditis. The most characteristic echocardiographic features of 12 patients with constrictive pericarditis were compared with the findings in 10 normal volunteers. Left ventricular posterior wall (LVPW) "flatness" was quantified by measuring the diastolic change in distance from the crystal artifact to the LVPW endocardium. In 11 of 12 patients the net diastolic LVPW endocardial movement was less than 1 mm. In 10 normal volunteers LVPW endocardium moved posteriorly in diastole from 1.5 to 4 mm (mean 2.2 +/- 0.8). Abnormal septal motion was present in five of 12 patients with constriction. Pericardial thickness measured using standard damping techniques for both cnstriction and normal population did not distinguish the two groups. The "flattening" of the left ventricular endocardium as quantified above verifies earlier qualitative observations and was the most consistent finding in this series of patients with constrictive pericarditis.


Aortic infarction following dissecting aortic aneurysm. Aortic infarction was observed in 21 of 34 cases of dissecting aortic aneurysm. This lesion occurred as a central zone of necrosis with preserved elastic laminae, sparing media adjacent to the true and false lumens. In cases where the false lumen was occluded, the central infarction extended to this lumen. The infarction followed rather than preceded dissection, took approximately 48 hours to develop, and did not organize with time. The lesion occurred exclusively in the thoracic aorta, and bore no relationship to medial cystic necrosis. Present surgical therapy does not extirpate these areas, and the implication of these lesions in terms of management remains to be determined.


ST segment changes post-infarction: predictive value for multivessel coronary disease and left ventricular aneurysm. To ascertain whether exercise testing might predict multivessel coronary disease and left ventricular aneurysm after a myocardial infarction, 154 patients with a single documented myocardial infarction who had both exercise testing and coronary angiography were grouped according to whether they had greater than or equal to 1 mm ST depression, greater than or equal to 1 mm ST elevation, or neither during exercise testing: 83 patients developed ST depression alone (group 1); 22 patients had ST elevation with concomitant ST depression in other leads (group 2); 19 patients had ST elevation alone (group 3); and 30 patients had no ST changes (group 4). Multivessel disease, defined as greater than or equal to 70% luminal narrowing in two or more coronary vessels, was present in 76% (63 of 83) of group 1, 91% (20 of 22) of group 2, 21% (four of 19) of group 3, and 13% (four of 30) of group 4. A left ventricular aneurysm was present in 31% (26 of 83) of group 1, 68% (15 of 22) of group 2, 79% (15 of 19) of group 3, and 40% (12 to 30) of group 1. We conclude that ST changes during exercise testing in patients after a myocardial infarction can reliably predict the extent of coronary disease and the presence of a left ventricular aneurysm; ST depression with or without ST elevation predicts multivessel disease; ST elevation alone or a negative exercise test suggests single vessel involvement; and ST elevation with or without ST depression predicts left ventricular aneurysm.


Platelet aggregation in aortic and coronary venous blood in patients with and without coronary disease. 3. Role of tachycardia stress and propranolol. We studied 16 patients with coronary artery disease (CAD) to evaluate platelet aggregation in blood samples withdrawn simultaneously from the aorta and coronary sinus. At rest, mean platelet aggregation in coronary venous blood was significantly lower than that in aortic blood. Platelet counts in coronary venous blood were also lower than in the aortic blood in each of the six CAD patients in whom counts were done. Platelet aggregation was lower in seven patients who were taking propranolol than in the remaining nine who were not taking propranolol. During tachycardia stress, platelet aggregation increased in all patients, but the magnitude of increase was greater in patients not taking propranolol. In four other patients without CAD, platelet aggregation and counts were also studied in the same fashion and were similar in both the aortic and coronary venous blood. These data suggest that in certain CAD patients, platelet consumption or destruction within atherosclerotic vasculature may occur. Propranolol may reduce platelet aggregation at rest and modify excessive aggregation during tachycardia stress in certain CAD patients.


Electrocardiographic precordial mapping in anterior myocardial infarction. The critical period for interventions as exemplified by methylprednisolone. Serial 72-point precordial mapping of ECG has been recorded to describe the natural history of changes in the precordial areas of ST segment elevation and the development of Q waves in 51 patients with acute uncomplicated anterior myocardial infarction. Eight patients have been studied in the same way but received 25 mg/kg of methylprednisolone sodium succinate as a single intravenous injection within 6 hours from the onset of chest pain. There was a linear relationship between the stable precordial area of Q waves at 24 hours and the rapidly changing precordial areas of ST segment elevation at 2--3 hours, 5--6 hours and 12 hours after the onset of pain in the untreated patients. When methylprednisolone was given, the treated patients developed a smaller precordial area of Q waves at 24 hours than was predicted from the precordial area of ST elevation recorded before the drug was given. This study has introduced a technique that can provide a qualitative assessment of the relationship between ECG evidence of ischemia and infarction in each patient.


Impedance plethysmography: the relationship between venous filling and sensitivity and specificity for proximal vein thrombosis. We investigated the hypothesis that the diagnostic accuracy of impedance plethysmography (IPG) for thrombosis of the popliteal or more proximal veins increases with enhanced venous filling. Venous filling was increased by prolonging cuff occlusion and by sequential testing. IPG and vengography were performed on 169 legs with and 317 legs without proximal vein thrombosis. The sensitivity and specificity of IPG rose significantly with increased venous filling. Changes in venous filling were associated with corresponding changes in emptying in normal legs, but not those with proximal vein thrombosis, so that the regression lines relating venous filling and emptying in normal and abnormal legs diverged significantly (P less than 0.001). If the IPG sequence had been terminated after only a single 45 second occlusion time test, sensitivity would have deteriorated by 10% and specificity by 20%. These observations indicate that the accuracy of IPG can be significantly enhanced if optimal venous filling is obtained.


Arterial insufficiency of the hand evaluated by digital blood pressure and arteriographic findings. In 80 hands with arterial occlusive disease, systolic blood pressure at the wrist and in all fingers was measured by photoelectric plethysmography. The correlation of such pressures with angiographic evidence of organic obstruction or the development of digital ischemic signs was studied. Digital blood pressure was normal in all 184 fingers in which there was at least one obstruction-free arterial path to and through the finger. Ischemic signs developed in only one. Of 203 fingers with occlusion in vessels in or leading to both sides of the finger, a decreased pressure was seen in 173 and a normal pressure in 30. Ischemic signs were observed in 132 of 173 fingers with a decreased pressure, and in two of 30 with a normal pressure. The clinical significance of measurement of digital blood pressure in arterial occlusive disease is discussed.


Stability of blood pressure rank and urinary kallikrein concentration in childhood: an eight-year follow-up. Previous studies in a population of 721 children aged 2--14 years demonstrated the familial aggregation of blood pressure in children, and a significant regression coefficient (b = 0.25) of follow-up on initial blood pressures over a four-year period. Urinary kallikrein concentration (UKal) also aggregated in families, was lower in black than in white children and was inversely related to blood pressure. Further studies in the same cohort have been conducted. These variables were again measured in 484 children in 129 families seven to eight years after the initial blood pressure and three to four years after the original UKal measurements were made. Familial aggregation again was found for blood pressure and urinary kallikrein. Blood pressure tracking was demonstrated by the finding that blood pressure scores at the third survey were related significantly to those at both previous surveys. Kallikrein concentrations in casual urines at Survey 3 were related to those obtained at Survey 2 (r = 0.499), and were again significantly lower in black than in white children (log = 3.84 +/- 0.8 vs 4.37 +/- 0.7; P less than 0.001). These were significant inverse relations between UKal/creatinine concentration and blood pressure in both white and black children. Thus, familial aggregation of blood pressure, blood pressure rank and concentration of kallikrein in casual urine specimes are relatively stable in children over an eight-year period of observation. This study demonstrates in a free living population of normal children, a stable relation between blood pressure and an enzyme which is involved in the production of potent vasodilator peptides and is related to hypertension in adults.


Cardiac manifestations of malignant hyperthermia susceptibility. Malignant hyperthermia is a disease resulting from defective cellular membranes, usually presenting as drug-induced pyrexic crises. We describe four patients with life threatening ventricular arrhythmias or chest pain in the absence of pyrexic crises. Three presented with life threatening arrhythmias and the fourth with severe atypical chest pain. Two patients had a family history of multiple sudden deaths. Resting CKs were elevated in three patients while CK-MB was elevated in one. Resting ECGs were abnormal in three. Three patients had recurrent ventricular tachycardia, two had recurrent ventricular fibrillation and multiple cardiac arrests. Cardiac catheterization showed abnormal left ventricular wall motion in two and minimal mitral valve prolapse in one while all had normal coronary arteries. Thallium-201 myocardial imaging demonstrated large perfusion defects in the patient with electrocardiographic Q waves and normal coronary arteries. Myocardial involvement has been demonstrated by clinical, electrocardiographic, hemodynamic, angiographic and myocardial imaging abnormalities. Malignant arrhythmias occurred in these patients in the absence of pyrexic crises or drug admininstration. Abnormal calcium release in the myocardium, as documented in skeletal muscle membranes, may be a unifying concept for the various manifestations described.


Radiation exposure to the operator performing cardiac angiography with U-arm systems. We measured the radiation exposure received by a group of operators performing 700 coronary angiograms. All studies were performed using the brachial artery approach and the Philips Cardio Diagnost. Nineteen sites were monitored on each operator, using lithium fluoride thermoluminescent dosimeters. Four hundred examinations were performed with a table-mounted protective shield in place. Three hundred were performed without the shield. The average exposures (in mR per study) with and without the shield were 1.9/6 for the eyes and 1.4/8.3 for the thyroid. The resulting operator exposure with the shield in place is low enough so that an operator performing 25 procedures per week on a continuous basis will not exceed the recommendations of the National Commission on Radiological Protection and Units. We therefore strongly recommend the use of properly designed and appropriately positioned shield with all U-arm systems.


Temporary, catheter-induced block in accessory pathways. Catheter-induced conduction delay or block in the accessory atrioventricular pathway of four patients with the preexcitation syndrome was observed. Block in the accessory pathway occurred during routine catheter placement and lasted from 90 seconds--14 hours. All of the pathways were in locations readily accessible to catheter trauma, located in the right anterior septal/paraseptal area in three patients, and in the posterior septum near the orifice of the coronary sinus in the fourth. The location of the acessory pathway was confirmed at surgery by endocardial mapping in three of the four patients. The fourth patient did not undergo survey. These findings suggest the need for caution in performing and interpreting electrode catheter studies which are undertaken to document the presence of accessory pathways.


Electrophysiological demonstration of concealed conduction in the human atrium. In the course of electrophysiological evaluation of six patients with sick sinus syndrome, two patients with chronic conduction system disease and four patients with paroxysmal supraventricular tachycardia or atrial flutter-fibrillation, the phenomenon of concealed conduction within the atrium (At) was repeatedly observed. One pair of intracardiac electrodes was used to deliver electrical stimulus (St) to the high right At and two additional pairs of electrodes were utilized to record high right and low septal right atrial electrograms. The interelectrode distances were 10 mm apart. In all 12 patients, high right atrial capture could be accomplished at a pacing rate of greater than or equal to 200 beats/min. Concealed intra-atrial conduction was evident when the intra-atrial conduction time of the propagated St during 2:1 St-At block was more than 40 msec longer than that during 1:1 St-At conduction at half the St frequency. This indicated that the nonpropagated St during 2:1 St-At block partially penetrated the At and in turn, delayed conduction of the subsequently propagated St. Futher observations revealed that a gradual increase in the St frequency resulted in a progressive prolongation of the intra-atrial conduction time and a shifting of 2:1 to 3:1 St-At block (alternating Wenckebach periodicity in the At) in five patients. These findings clearly demonstrate the occurrence of concealed conduction in the human At.


Fibrolipoma of the mitral valve in a child. Clinical and echocardiographic features. Echocardiographic examination, performed in a 12-year-old boy who had signs of mitral regulation, showed the presence of an abnormal mass of echoes in the left atrium and mitral orifice. These were initially interpreted as representing an atrial myxoma. Surgical exploration showed that the tumor originated from the posterior leaflet of the mitral valve, and, microscopically, the lesion was diagnosed as a fibrolipoma. The clinical and echocardiographic features of this unique type of cardiac tumor are discussed.


Automated metabolic profiling of organic acids in human urine. I. Description of methods. We describe a complete procedure for separation and mechanized analysis of organic acids in human urine. The acid fraction of urine is separated by anion-exchange chromatography on diethylaminoethyl-Sephadex. Individual acids are identified and measured by use of a gas-chromatography/mass spectrometer/computer system that can clearly distinguish contributions from at least 150 substances in a single sample. We discuss analytical recoveries, contributions from the sample separation process, stability of stored samples before and after processing, and reproducibility of the extraction procedure.


Automated metabolic profiling of organic acids in human urine. II. Analysis of urine samples from "healthy" adults, sick children, and children with neuroblastoma. Normalized median, minimum, and maximum values (analytical concentration factors) are given for 134 organic acids in urine of nine adult control subjects, five juvenile control subjects, and five children with neuroblastoma. The organic acids, separated by anion-exchange chromatography, were analyzed by a gas chromatograph-mass spectrometer-computer system. Sixty substances in this fraction are positively identified, and, of these, mean absolute concentrations are listed for 20. An additional 81 substances, sought but not found by this method, and 16 other substances found in a subset of these urines by another analytical method, are also listed. Measured retention indices on 5% OV-17 and a selected discriminating ion are given for each of the total of 231 compounds. Results are compared for the three groups of subjects, and the value of normalizing the data is discussed.


Simple method for estimating glycosylated hemoglobins, and its application to evaluation of diabetic patients. Recent reports have suggested that determination of glycosylated hemoglobin may serve as a clinical aid for long-term blood glucose control in diabetes mellitus. We describe a simple procedure for measuring it by ion-exchange chromatography. Hemolysates were subjected to Bio-Rex 70 chromatographic separation on small columns. Percentages in the normal group ranged from 4.7 to 8.8% of total hemoglobin; the mean +/- standard error was 6.61 +/- 0.31%. Values in the diabetic group ranged from 6.9 to 17.4%; the mean was 10.83 +/- 0.34. Plasma glucose concentrations after fasting, plotted vs. the percent of glycosylated hemoglobin, revealed a linear relationship at normal or moderately high glucose concentrations. However, the values for glycosylated hemolgobin approached a plateau with grossly higher plasma glucose concentrations after fasting. Our results support the view that, due to its long half-life, the estimation of glycoylated hemoglobin reflects the integrated glucose concentrations to which the erythrocytes have been previously exposed.


Evaluation of a glassy-carbon electrode for amperometric detection of selected methylxanthines in serum after their separation by reversed-phase chromatography. We examined the relative merits of direct-current, pulse, and differential pulse amperometry at a flow-through, glassy-carbon electrode used in monitoring column chromatography. We give a rapid, simple procedure for determining theophylline (1,3-dimethylxanthine) in 20--100 microliter of blood serum. Interference by 1,7-dimethylxanthine, a metabolite of caffeine, is compared for photometric and amperometric detection, and some advantages of combined photometric and amperometric detection are discussed.


Negative-ion mass spectrometry of amphetamine congeners. Negative-ion chemical ionization mass spectrometry with a conventional combined gas chromatograph-mass spectrometer has been used for the analysis of amphetamine-like compounds. Ammonia was used as the reagent gas, which gives rise to few but specific sample-fragment ions, such as (M--1)- as the base peak, m/z 91, and a smaller peak corresponding to the end part of the side chain. Five reference compounds and a urine sample from an overdose case were analyzed. Comparative positive-ion chemical ionization reference spectra were also recorded.


Enzymic fluorometric continuous-flow assays for blood glucose, lactate, pyruvate, alanine, glycerol, and 3-hydroxybutyrate. We describe enzymic fluorometric methods of automated analysis for glucose, lactate, pyruvate, 3-hydroxybutyrate, glycerol, and alanine in perchloric acid extracts of blood. Unmodified Technicon AutoAnalyzer II apparatus is used. The usual concentrations of all these metabolites can be measured in as little as 0.1 ml of blood from a fasting subject. Within-batch and between-batch coefficients of variation ranged from 0.4 to 4.4% for all metabolites except 3-hydroxybutyrate, for which CV's were higher for low concentrations. Analytical recovery of added metabolites ranged from 92 to 98%. Glucose, lactate, alanine, and 3-hydroxybutyrate are stable in perchloric acid extracts for at least 13 days at room temperature, and a year at -20 degrees C; pyruvate shows a 6--8% loss after 3 days and 52% by one year at -20 degrees C; glycerol concentrations were stable at -20 degrees C for at least 13 days. Blank fluorescence is found in perchloric acid extracts of blood, necessitating blank runs for pyruvate, 3-hydroxybutyrate, glycerol, and alanine. The systems are simple to use, relatively inexpensive to operate, and are recommended for any laboratory with high throughput of samples.


Enzymic micro-assay for blood glutamine. Blood glutamine is hydrolyzed by glutaminase; the resulting ammonia is dialyzed out and measured by use of a continuous-flow colorimetric microassay. Concentrations in the range of 10 to 1300 micromol of glutamine per liter can be determined in a 40-microliter sample. With slight modifications, higher concentrations, up to 5 mmol/liter, can be measured. Assay conditions and the qualities of the microtechnique were made optimal. Usual values obtained with this method for arterial and venous blood in man are, respectively, 443 +/- 140 and 466 +/- 142 micromol/liter (mean +/- 2 SD). Corresponding values for rats are 320 +/- 92 and 340 +/- 108 micromol/liter. Fasting causes a decreased blood glutamine.


Measurement of three aminoglycosiee antibiotics with a single radioimmunoassay system. A versatile radioimmunoassay procedure has been developed to specifically assay gentamicin, tobramycin, or amikacin. Iodinated derivatives of these three antibiotics were prepared by a modified Bolton and Hunter procedure. A single reagent system composed of a pool of three very specific antisera was used in the procedure. The antisera combination did not alter the specificity of the procedure. Validation of the assay of the three antibiotics included studies of parallelism, accuracy, precision, reproducibility, and method comparison with other radioimmunoassay procedures involving single antiserum systems. We discuss the application of this multiple assay capability to other procedures involving use of immunochemicals.


Thyroid hormone contamination in crystalline diiodotyrosine. Crystalline preparations of diiodotyrosine contain trace quantities of thyroxine that can be detected by several competitive radioassays and by chromatography on Sephadex. Spontaneous coupling of diiodotyrosine to thyroxine during iodination of tyrosine and coprecipitation of hormone with diiodotyrosine crystals are likely explanations for the trace contamination. We discuss the significance of these findings in relation to studies on the biosynthesis of thyroxine.


High-performance liquid-chromatographic determination of free nicotinic acid and its metabolite, nicotinuric acid, in plasma and urine. We report a liquid-chromatographic procedure for determining free nicotinic acid and a metabolite, nicotinuric acid, in plasma and urine. Five-tenths milliliter of urine or deproteinized plasma is evaporated and the residue analyzed isocratically by reversed-phase ion-pair chromatography, with measurement of the eluted nicotinic acid and nicotinuric acid at 254 nm. Nicotinic acid, nicotinuric acid, and the internal standard (isonicotinic acid) have retention times of 7.8, 8.4, and 6.8 min, respectively, in plasma, and 12.3, 13.1, and 10.8 min in urine, because of double column length. Day-to-day reproducibilities (CV) for nicotinic acid and nicotinuric acid within 7.5% are attainable for the concentration ranges 0.1--20 mg/liter, equivalent to 0.81--162 micromol of nicotinic acid and 0.55--11 micromol of nicotinuric acid per liter for plasma; in urine for the range 0.5--100 mg/liter, equivalent to 4--810 micromol of nicotinic acid and 2.8--555 micromol of nicotinuric acid per liter. Metabolites of nicotinic acid such as nicotinamide, N-methylnicotinamide, 2-hydroxypyridine-5-carboxylic acid, and other structurally related substances do not interfere.


Determination of a trivariate reference region for free thyroxine index, free triiodothyronine index, and thyrotropin from results obtained in a health survey of middle-aged women. We measured free thyroxine index, free triiodothyronine index, and thyrotropin in serum in a community survey of the female population 39--60 years old. A statistical method using Mahalanobis' distance was applied to the data, to identify and eliminate highly abnormal values, "outliers." There was a small but statistically significant increase in each of the three hormones with age. After correction for age dependency, significant but rather small correlations were found between the hormones. The material was used to calculate univariate reference intervals and a trivariate 0.95-tolerance region.


Modified erythrocyte uroporphyrinogen I synthase assay, and its clinical interpretation. Assay of erythrocyte uroporphyrinogen I synthase is an accepted diagnostic test for acute intermittent porphyria, particularly in those individuals who are asymptomatic or in whom the disease is not biochemically manifested by excretion of excess porphyrin precursor. The assay described is based upon a coupled-enzyme procedure in which added delta-aminolevulinic acid and its dehydratase present in erythrocytes are used to generate porphobilinogen as substrate for uroporphyrinogen synthase. Zinc and dithiothreitol are added with preincubation to give maximum activity and reproducibility. These agents also prevent inhibition by lead. Healthy young women had a mean activity of 40 nmol of porphyrin formed per milliliter of erythrocytes per hour, men and activity of 38 nmol/ml/h. Preparation of control specimens is described. This assay gave within-day CVs ranging from 1.9 to 2.8%. Precautions in interpretation of results are discussed.


Neonatal thyroxine screening by use of a single-tube solid-phase radioimmunoassay. Filter paper discs saturated with dried blood can be used in the Immunotube solid-phase thyroxine radioimmunoassay. This assay utilizes polypropylene tubes to which antibody to thyroxine is covalently bound. The filter paper standards and samples are placed in the tubes, followed by an assay buffer that contains I125-labeled thyroxine and compounds to displace thyroxine from its binding proteins. After incubation, bound and free thyroxine are separated by aspirating or decanting the disc and buffer from the tube. The test can be used with 0.32 cm (1/8 inch) or 0.64 cm(1/4 inch) discs, and gives quantitative results that correlate well with those for serum samples. The intra-assay coefficient of variation is less than 10%. The assay may readily be mechanized with existing disc-punching equipment, and results of its use in mass screening programs are described.


Increased D-glucaric acid excretion by jaundiced patients. The urinary excretion of D-glucaric acid, a catabolite of glucuronic acid, is considered to be a reliable index of the state of hepatic microsomal enzyme activity. Because enzyme activity may be altered in liver disease, we examined the effect of liver disease on the excretion of this metabolite and its correlation with liver function tests. We studied 89 patients with nonhemolytic jaundice, 39 with viral hepatitis, 33 with obstructive jaundice, six with cirrhosis, and 11 patients with jaundice of mixed etiology. Glucaric acid excretion was significantly increased in all these patients as compared to controls, most pronounced in the obstructive jaundice group. No correlation was found between glucaric acid excretion and concentrations of bilirubin, albumin, globulin, aspartate aminotransferase, alkaline phosphatase, cholesterol, or gamma-glutamyltransferase in serum, even though the concentrations of these analytes did vary with the type of liver disease. We suggest that this increase in glucaric acid excretion is an indication of normal or even increased glucuronidation (UDP-glucuronosyltransferase activity), which occurs in liver disease.


Is serum cholinesterase activity a predictor of succinyl choline sensitivity? An assessment of four methods. Four methods for measuring serum cholinesterase activity have been applied to sera of normal individuals and of patients shown to be sensitive to short-acting muscle relaxants of the succinyldicholine type. They have been assessed according to their ability to differentiate between sensitive and insensitive individuals on the basis of enzyme activity measurements alone. The method described, based upon that of Dietz et al. [Clin. Chem. 19, 1309 (1973)], in which propionylthiocholine is used as substrate, is best for this purpose, being capable of identifying over 90% of affected individuals with no false positives. Acetylcholine and butyrylthiocholine are slightly inferior substrates in this respect, and benzoylcholine gives little useful information.


Use of polyethylene glycol in radioimmunoassay of human placental lactogen. We describe a rapid radioimmunoassay for human placental lactogen in biological fluids, with use of polyethylene glycol to separate free from antibody-bound placental lactogen. The standard curve obtained, easily fitted to a logit-log transformation, is useful over a concentration range of 6 to 400 ug/liter. Within-assay variability is 1.97%, between-assay variability 2.20%. The relation is linear when lactogen added is plotted against that accounted for analytically, the actual recovery being 100.26%.


Improved micromethod for determination of underivatized clonazepam in serum by gas chromatography. We describe a gas-chromatographic micromethod, with use of a solid injection system and an electron capture detector, for determination of underivatized clonazepam in serum. Vigorous mixing of 100 microliter of serum with 20 microliter of borate buffer (pH 9.0) and 100 microliter of a cyclohexane/dichloromethane mixture (4/6 by vol) containing methyl nitrazepam as internal standard suffices to extract 86% of the clonazepam into the organic layer. We obtained a linear response curve for clonazepam in serum in the concentration range 5 to 150 microgram/liter. The lowest detectable concentration by our method is about 1 microgram/liter of serum. Interference by several anticonvulsant drugs, metabolites, and related compounds was investigated. The assay is simple, rapid, and suitable for clinical routine determination of clonazepam in serum in therapeutic concentrations. We illustrate the steady-state concentration of clonazepam in serum during chronic oral administration to 28 patients.


Effects of bilirubin on detection of hydrogen peroxide by use of peroxidase. Bilitubin interferes with the quantitation of hydrogen peroxide in reagent systems in which peroxidase is used. Difference spectra suggest that this interference is a combination of chemical and spectral effects. The data presented are most consistent with the following interpretation: (a) bilirubin destroys part of the reactive intermediate formed in the peroxidase reaction and thus decreases the amount of chromophore formed and (b) the spectra for bilirubin and the chromaphore overlap, which also affects the results. The relevance of these results to reagent design and laboratory quality control are discussed.


A method of assessing serum triiodothyronine concentrations that is independent of subject's age and variations in concentrations of binding proteins in serum. Pairs of values for serum triiodothyronine and serum triiodothyronine binding capacity were represented as maps of points for euthyroid populations (including subjects who were pregnant or were receiving estrogens) in the age groups 18-50, 51-64, 65-79, and greater than 80 years (504 subjects) and thyrotoxic populations in the same age groups (207 subjects). For each age group the two populations were effectively separated bythe technique. Statistical analysis by the method of principal components justified the combination of all the age-related maps into a single map, allowing the assessment of serum triiodothyronine regardless of age or concentration of binding proteins in serum. The overlap of the combined populations was less than 1%. The method proposed is superior to a calculation of free triiodothyronine index in several respects.


Immobilized-enzyme nylon-tube reactor for routine determination of uric acid in serum. We report here the preparation of an immobilized-enzyme nylon-tube reactor containing uricase (EC 1.7.3.3) and the assembly of a flow-through system (Technicon AutoAnalyzer II) for the routine determination of uric acid in serum. Results of these uric acid analyses by use of immobilized uricase, in conjunction with peroxidase (EC 1.11.1.7) and aminophenazone-dichlorophenol in solution, are compared with those obtained with the same enzyme in solution by use of the uricase-PAP (peroxidase, 4-aminophenazone, dichlorophenol) method. Clinical trials carried out routinely with the uricase reactor give reliable and reproducible results with high precision at an appreciably lower cost. The reactors are stable to continued or intermittent use for at least three months or for 4000 tests.


Increased creatine kinase isoenzyme MB values in patients without myocardial infarct. Six of 13 randomly selected patients in a medical intensive-care unit with above-normal creatine kinase MB activities had diagnoses other than myocardial infarction. These data, which indicate the need for further study, were obtained during evaluation of a commercially available column procedure (Biodynamics/bmc).


Gas--liquid chromatographic microdetermination of underivatized ethosuximide (alpha-ethyl-alpha-methyl succinimide) in plasma or serum. A gas-liquid chromatographic procedure for the microdetermination of ethosuximide is described. Ethosuximide is extracted from acidified plasma or serum into chloroform containing an internal standard alpha,alpha-dimethyl-beta-methyl succinimide. Part of the initial chloroform extract is gently evaporated, the residue redissolved in n-heptane at 60 degrees C, and an aliquot analyzed by gas-liquid chromatography. The procedure is rapid, reliable, sensitive, and specific. It requires a 25--50 microliter sample for a single estimation, has a detection threshold of less than 10 micromol/liter, and is suitable for routine clinical use.


Distribution of thioridazine and its metabolites in human tissues and fluids obtained postmortem. Thioridazine and metabolites were measured in brain, liver, and kidney specimens, obtained postmortem from two subjects whose deaths were related to acute intoxication with thioridazine, by gas-liquid chromatography. Although the absolute concentration measured for thioridazine and metabolites differed in the two cases, the metabolic pattern for each tissue, expressed in terms of the percentage of total drug in each tissue, was quite similar. The brain, liver, and kidney metabolic patterns, however, are in sharp contrast to the plasma metabolite patterns observed for subjects on a therapeutic regimen of thioridazine. As this example demonstrates, postmortem specimens are a valuable (but seldom used) source of human pharmacological data.


Plasma estriol vs. estrogen assays in 24-h urines as an index to fetal status. Conventionally, urinary estrogens are measured to monitor fetal well-being in the third trimester of pregnancy. There are many theoretical and practical advantages to the use of recently introduced plasma estriol assays; consequently, we undertook to compare the results of a spectrophotometric assay for total urinary estrogen with values for plasma total and free (unconjugated) estriol obtained by radioimmunoassay with commercially available kits. The data correlate well, r = 0.70 for plasma total estriol with urinary estrogen (expressed as milligrams of estrogens per gram of creatinine [E/C]), r = 0.79 for plasma free estriol with E/C, and r = 0.71 between the two plasma values. Minute-to-minute fluctuations during 90 min and day-to-day fluctuations followed during five days suggest that fluctuations of up to +/- 45% are within the normal range (means +/- SD of the day-to-day fluctuations were 16.9 +/- 7.7% and 15.6+/- 8.2%). Turn-around time is a minimum of 29 h for the urinary estrogen assay, 4.5 h for the plasma total estriol, and 3 h for the plasma free estriol.


Reduced-volume radioimmunoassays for parathyrin and calcitonin in serum, for use in pediatric and small-animal studies. Radioimmunoassays for circulating parathyrin and calcitonin used in most laboratories require 0.6--0.7 ml of serum or plasma, or more, for assay of one hormone. Such volumes are often difficult to obtain safely or repeatedly from pediatric patients or in small-animal studies. We modified our existing procedures to markedly decrease sample and reagent requirements. All reagent volumes were proportionately reduced to one-fourth the usual (from 500 microliter final incubation volume to 125 microliter), and serum or plasma volumes from 200, 100, and 50 microliter to 50, 25, and 10 microliter. We used smaller (10.3 X 50 mm) tubes and slightly modified the separation with charcoal. Results, validated by studies in rats, showed the sensitivity to match that of the usual assays; inter- and intra-assay variance was less than 20%. Simultaneous regular- and reduced-volume assays of parathyrin in sera from 19 children gave almost identical results (r = 0.9987). Both hormones can be assayed in less than 400 microliter of serum.


Alternative mechanized method for analysis for inorganic phosphate without reduction. The mechanized measurement of inorganic phosphate in plasma can be achieved without reduction by only minor alterations to an existing continuous-flow reduction procedure. The method is based on the enhancement of the extinction of the phosphomolybdate complex by the detergent Triton X-100. Results obtained agree well with those obtained by the standard reduction (Fiske-SubbaRow) procedure. The correlation coefficient between the two methods was 0.996 and statistical analysis of the data confirmed very good agreement between the two sets of values.


Modified Jendrassik--Grof method for bilirubins adapted to the Abbott Bichromatic Analyzer. I describe a semi-micro method for use with a discrete analyzer, the Abbott ABA-100, requiring 75 microliter of sample for determination of both the total and direct bilirubins. This method utilizes a serum blank. It is simple and reasonably rapid. The azobilirubin formed by both the total and the direct bilirubins produces a reddish-violet color, which is measured at 550 nm. Values obtained are comparable to those by the automated Jendrassik and Grof procedure adapted by Gambino and Schreiber (total r = 0.999, direct r = 0.990). The method is linear to 300 mg/liter. Day-to-day precision (CV) for the total bilirubin was 8.8% for a 8 mg/liter sample (n = 45) and 4.7% for a 57 mg/liter sample (n = 46). The absorbance of hemoglobin and its derivatives (540 nm) caused a false decrease in the measured bilirubin content, the direct being more affected than the total. This problem was alleviated by the use of caffeine and acid blanks.


Interlaboratory comparison of lead and cadmium in blood, urine, and aqueous solutions. Analysis for lead and cadmium in biological liquids (blood and urine) is difficult. Results of such analyses from five laboratories are compared for samples with known additions of lead and cadmium. The data, evaluated in terms of inter- and intralaboratory reproducibility and accuracy, suggest that laboratories should voluntarily participate in quality control programs. Users of routine laboratories are advised to use their own quality control program.


Evaluation of the discriminative power of lipidophoresis by discriminant analysis using a simple quantitative method of agarose lipidophoresis. A simple, reproducible and quantitative method for separation of lipoproteins in agarose is presented. The validity of the results is shown by comparison with gravimetric analysis of fractions separated by ultracentrifugation in a discontinuous NaCl-KBr gradient. Results for a large group of normals and of patients with various lipid disorders are presented, and discriminant analysis is used to evaluate objectively the potential of lipidophoresis in the classification of hyperlipoproteinemias. It is shown that quantitative lipidophoresis gives as much information as the determination of cholesterol and triglycerides, for classification purposes.


A simplified assay for methotrexate in biological fluids using a crude yeast lysate. Routine pharmacokinetic monitoring of methotrexate levels in biological fluids appears to be important for clinical applications involving large doses of methotrexate with leucovorin rescue. Recently several laboratories have independently described a competitive radiobinding assay that has the necessary sensitivity, precision and simplicity for this purpose. This study describes a modification of this assay method using a lysate of dry, active yeast as a source of binding activity. The assay is sensitive at blood concentrations of about 1 nM. No significant interference can be shown with naturally occurring folates. Specificity has also been shown by recovery experiments from serum and urine. Thus this modified assay matches the previously described radiobinding assays but with the advantage of using a methotrexate binding reagent that is readily available and quite inexpensive.


Rapid colorimetric methods for quantitative lipoprotein-X determination without electrophoresis. Two procedures for quantitative determination of the cholestasis-characterizing lipoprotein-X are described. Both procedures are based on ether-extract-ability of phospholipids which are relatively loosely bound in lipoprotein-X. The presence of cetyltrimethylammonium bromide at adequate concentration allowed complete ether extraction of phospholipids from lipoprotein-X even at low concentration, without affecting the phospholipids of normal lipoproteins. In one modification phospholipids were extracted into ether ion-paired with a bilirubin conjugate (e.g. Woodward's K conjugate) the concentration of which was then estimated in the ether extract. In the second modification phospholipids were directly determined in the ether extract. Calibration was done either with a lipoprotein-X concentrate or with a clear sonicated lecithin sol added to normal serum. The results obtained by both methods are in a good agreement with the results by agar gel electrophoresis.


Metal-binding studies of metallothioneins in Menkes kinky hair disease. Two species of metallothioneins were isolated from both normal and Menkes kinky hair disease (MKHD) patient livers. Atomic absorption determination of metals indicated that the patient liver metallothioneins had lower copper and cadmium content than normals. Isotope exchange studies, carried out by incubating native metallothioneins with copper-64 or cadmium-109 demonstrated a decreased affinity for copper and an increased affinity for cadmium in both MKHD metallothioneins. An hypothesis is proposed in which metallothionein functions as an intracellular copper carrier and is responsible for the transport of copper between the cells and the surrounding. Change in the copper affinity of the metallothioneins was suggested to be the major abnormality in MKHD.


Use of a chromogenic substrate for the diagnosis of Krabbe's disease, with special reference to its application in prenatal diagnosis. A chromogenic substrate, 2-hexadecanoylamino-4-nitrophenyl-beta-D-galactopyranoside, has recently been described for the diagnosis of Krabbe's disease. Hydrolysis of this substrate by extracts of cultured cells and tissues was compared with the activities of lactocerebrosidase I and non-specific beta-galactosidase. Under appropriate conditions, hydrolysis of the chromogenic analogue was markedly reduced in extracts of cultured amniotic fluid cells and skin fibroblasts derived from cases of Krabbe's disease. Activity was also markedly deficient in extracts of Krabbe's brain, although only a partial reduction was measured in liver extracts. Generally activities were higher in tissues of fetal origion. Unfortunately, the new analogue proved less specific and less sensitive than the natural substrates used to diagnose Krabbe's disease. Consequently, the analogue does not provide a satisfactory alternative substrate for the prenatal diagnosis of Krabbe's disease.


Human hexosaminidase isozymes. Assay of platelet activity for heterozygote identification during pregnancy. Identification of carriers of the Tay-Sachs gene during pregnancy is difficult because of the increase in serum of a heat stable hexosaminidase isozyme I (or P) as well as changes in the relative and absolute activities of the various molecular forms of the enzyme with advancing pregnancy. In contrast, isolation of blood platelets followed by ion exchange chromatographic separation and assay of the hexosaminidase isozymes in platelet extracts by an automated method provides a sensitive and reliable method for heterozygote identification during pregnancy. This method appears superior to procedures involving thermal inactivation of extracts of peripheral blood leukocytes because of significant differences in the content of the hexosaminidase isozymes in granulocytes, lymphocytes and other cell types, as well as variations in the proportion of these cell types in samples of peripheral blood. It also alleviates the problem inherent in any method involving thermal inactivation of hexosaminidase A by avoiding possible interconversion of the various molecular forms of the enzyme associated with heating.


Studies on glutaryl-CoA dehydrogenase in leucocytes, fibroblasts and amniotic fluid cells. The normal enzyme and the mutant form in patients with glutaric aciduria. Three patients with glutaric aciduria have been shown to possess a partial but severe defect of the enzyme glutaryl-CoA dehydrogenase in isolated leucocytes and cultured skin fibroblasts. They could readily be distinguished from heterozygotes by measuring the activity of this enzyme, as shown in a study of the two families involved. The activity of glutaryl-CoA dehydrogenase in normal cultured amniotic fluid cells was comparable to the activity in normal cultured skin fibroblasts indicating the possibility of prenatal diagnosis. Without flavin adenine dinucleotide added to the assay mixture, the activity of glutaryl-CoA dehydrogenase in fibroblasts from normal individuals was very much reduced and similar to the activity in the patients, but after addition of flavin adenine dinucleotide to saturation the activity increased 20-fold in normal subjects while only a very slight increase could be demonstrated in the patients. The Michaelis constant for the substrate glutaryl-CoA was similar for both normal and patient cell lines. The optimum assay conditions for the enzyme in cultured fibroblasts from normal individuals have been established. In contrast to our patients, we found no activity in a fibroblast cell line from a patient with glutaric aciduria diagnosed elsewhere.


Serum methionine: its rapid determination by continuous-flow colorimetry. A rapid automated method for the determination of serum methionine is presented for use with the Technicon AutoAnalyzer system. It is based on the decolorisation of halide platinates by organic sulphides, including methionine. The 5--95% reference range from 249 sera from hospital patients (excluding those with liver and renal diseases and diabetes) was 20--60 mumol/l, mean 35 mumol/l. The S.D. of the method for 40 duplicate samples was 2.2 mumol/l. Recoveries ranged from 101--105%. This measurement facilitates the clinical evaluation of liver disease, especially acute liver failure, kidney disease and diabetes mellitus.


Correlations between branched-chain amino acids and branched-chain alpha-keto acids in blood in maple syrup urine disease. In 62 blood samples from 3 patients with classical maple syrup urine disease and from one patient with a variant form, a close linear correlation was found between levels of branched chain amino acids and their corresponding alpha-keto acids. Keto acids were determined as O-trimethylsilyl quinoxalinols by gas chromatography with a nitrogen-selective detector.


Gas chromatographic determination of hydroxyproline in urine hydrolysates. A simple and specific method for the determination of hydroxyproline in urine hydrolysates has been described. Hydroxyproline was converted into its N-isobutyloxycarbonyl methyl ester derivative without elaborate cleanup, which was analyzed by gas chromatography. Hydroxyproline was clearly separated from other urinary constituents on a 0.60% FFAP on dimethyldichlorosilane-treated Gas-Chrom P column. Kainic acid was used as the most convenient internal standard available. The relative standard deviations of peak height ratios were 1.15--2.51% at the 10--150 microgram levels. Percent recoveries of hydroxyproline added to urine hydrolysates ranged from 98.8 to 107.3%.


A placental type of membrane-bound arylamidase in serum and ascites from a patient with renal cell carcinoma. A placental type of membrane-bound arylamidase was detected in serum and ascites from a patient with renal cell carcinoma by isoelectric focusing. The enzyme had the same molecular weight, electrophoretic mobility on polyacrylamide gel, isoelectric point and KM value as placental membrane-bound arylamidase. The new type of membrane-bound arylamidase was named Shiba-isoenzyme after the patient's name.


Serum monoamine oxidase assayed with a new synthetic benzylamine derivative as substrate. Its clinical significance with special reference to hepatic fibrosis. Serum levels of monoamine oxidase were determined in 171 healthy adults and in 77 patients with hepatic diseases by use of a new synthetic derivative of benzylamine, 1-[(4-aminomethylphenyl)azo]-2-naphthol hydrochloride, as substrate. This new compound is more effectively oxidized by serum monoamine oxidase than by benzylamine, currently the most widely used substrate. Diagnosis of hepatic disease was confirmed in all patients by liver histology. The results suggest that the level of this enzyme in serum increases in parallel with the development of hepatic fibrosis.


A rapid method for the determination of volatile fatty acids in biological fluids. A rapid and simple method for the determination of volatile fatty acids in plasma and urine without any pretreatment is described. The gas chromatographic method reported here allows us to detect and quantitate in approx. 20 min biological VFA characteristics of various metabolic diseases. Conditions used are very mild so to avoid as much as possible any thermal decomposition of biological compounds.


[Potentiometric determination of hepatitis B surface antigen in biological fluids (author's transl)]. The present report describes a sensitive and reliable method for potentiometric determination of hepatitis B surface antigen in biological fluids, as an application of enzyme-linked immunoassay technique by a "sandwich" procedure. Specific antibodies are immobilized on a gelatine membrane which covers the sensor of an iodide-sensitive electrode. After immersion of the modified electrode in a dilute solution of antigen, the enzymatic activity is evaluated in a solution of peroxidase-labelled antibodies, in the presence of substrate and iodides. The electrode potential is a function of antigen concentration in the solution. This new procedure should find its application in the determination of substances present in very low concentration in biological fluids.


Effect of snap freezing, dithiothreitol and storage on estimations of estrogen receptor sites. Estrogen receptor assays were performed on human myometrium in order to determine optimal conditions for certain steps of the assay. The findings indicate that: (1) tissue should be snap-frozen in liquid nitrogen prior to assay, (2) dithiothreitol does not increase measurable binding site numbers, (3) estrogen receptors in the cytosol fraction are unstable and should be stored in excess of 1 day; (4) if storage prior to receptor assay is unavoidable, tissue should be stored at -20 degrees C or -70 degrees C for no longer than 1 week.


A practical proposal for arginine-vasopressin radioimmunoassay. This work proposes the use of (a) a commercially available homologous system AVP antibody-AVP (Arginine Vasopressin) standard and (b) new acquisitions for the improvement of sensitivity for AVP radioimmunoassay by separate or simultaneous use of two-phase sequential incubation and epsilon-aminocaproic acid (EACA). The antiserum used in the system described is very specific since none even cross-reacted with lysine vasopressin (LVP) and has an apparent affinity constant (K) of 0.909 +/- 0.047 X 10(12) l/mol. This is sufficiently high to detect 0.5 +/- 0.2 pg/tube, which is theoretically expected of biological AVP. The total assay time is less than 48 h.


Human diploid lung fibroblast cell lines WI 26 and WI 38 exhibit isozyme shift of alkaline phosphatase after viral transformation. The testing of the type of membrane associated alkaline phosphatase (EC 3.1.3.1) by immunotitration has revealed that there is a shift from a liver-like type of alkaline phosphatase in normal cell cultures of the human diploid fibroblast cell strains WI 26 and WI 38 to a placenta-like variant in cultures of the same cell strains after the transformation by the DNA-tumor virus SV 40, the WI 26 SV 40 and the WI 38 SV 40 cell lines. The immunologically detectable switch-over has been confirmed by measuring the apparent Michaelis constant and the heat stability of the AP from normal and transformed cells. Liver-like AP is heat labile and has an apparent Michaelis constant for p-nitrophenylphosphate (about 4.0 X 10(-4) M). The placenta-like AP shows heat stability and a lower apparent KM (about 2.2 X 10(-4) M). The appearance of the so-called Regan enzyme of AP in some human tumors in vivo is discussed in this connection.


Improved determination of beta-glucuronidase in serum. With phenolphthalein beta-glucuronide as the substrate in the serum beta-glucuronidase assay, centrifugation at high speed after addition of alkali was required to minimize blanks. Optimal substrate concentration was 3--6 mM. Excess substrate was inhibitory.


The quantitative determination of 5-S-cysteinyldopa and dopa in normal serum and in serum from patients with malignant melanoma by means of high-pressure liquid chromatography. A method is described for quantitative determination of 5-S-cysteinyldopa and Dopa in serum involving high-pressure liquid chromatographic analysis (HPLC) and electrochemical detection. The chromatographic system allows estimation of injected amounts corresponding to 25 pg of 5-S-cysteinyldopa and Dopa. In normal subjects the mean serum 5-S-cysteinyldopa concentration was 2.8 ng/ml (range 0.4--12 ng/ml) and the mean serum Dopa 6.3 ng/ml (range 4--10 ng/ml). Patients with melanoma metastases showed increased serum concentrations of 5-S-cysteinyldopa.


A granulocyte test for detection of cattle heterozygous for mannosidosis. alpha-Mannosidase and beta-N-acetylglucosaminidase activities were found to differ between lymphocytes and granulocytes isolated from bovine blood. Activities of both enzymes in granulocyte preparations were found to be related to the eosinophil content of the preparations. A procedure is described that allows definition of the mannosidosis genotype of adult cattle by giving consideration to the influence of eosinophil content of granulocyte preparations upon the activity of alpha-mannosidase relative to that of beta-N-acetylglucosaminidase.


Protein kinase and adenylate cyclase of erythrocyte membrane from patients with Duchenne muscular dystrophy. In freshly prepared erythrocyte membranes from normal individuals and from patients with Duchenne progressive muscular dystrophy the endogenous protein kinase and the cAMP stimulated phosphorylation was identical for the three main 32P proteins including spectrin (protein band II). Another enzyme, adenylate cyclase, was found unchanged. Altered protein kinase and adenylate cyclase has been reported in this disorder. We have no explanation for these discrepancies.


Isoelectric focusing of acid hydrolases in human liver and serum. Findings in sera from one patient with I-cell disease phenotype. Isoelectric focusing was performed with six acid hydrolases in serum and liver tissue. Neuraminidase-treated serum and liver tissue were also examined. In general, acid hydrolases in sera seemed to be more sialylated than those in liver tissue. Serum from one patient with I-cell disease with increased activity of some acid hydrolases was found to have a normal isoelectric focusing pattern. These findings are discussed with respect to the uptake mechanism of acid hydrolases in serum.


The effect of lipoprotein lipase and hepatic lipase on the electrophoretic mobility of lipoprotein-X. Lipoprotein-X containing plasma from a patient with familial lecithin:cholesterol acyltransferase (LCAT) deficiency, was used as substrate and incubated with postheparin plasma or partly purified lipases. LP-X could not be demonstrated by agar gel electrophoresis after incubation with postheparin plasma from a healthy subject, from a patient with chronic active hepatitis deficient in hepatic lipase, or with partly purified lipoprotein lipase. After incubation a marked increase in free fatty acids (FFA) was observed. In contrast LP-X was still present after incubation when postheparin plasma deficient in lipoprotein lipase or partly purified hepatic lipase was added to the substrate. Only minor changes in the concentration of FFA occurred. After addition of oleic acid to the substrate LP-X could not be demonstrated by agar gel electrophoresis. However, in the isolated low density lipoproteins, LP-X like particles were still present as viewed by electron microscopy. Our results strongly suggest that the change in electrophoretic mobility of LP-X was induced by the release of FFA. This was achieved by lipoprotein lipase, but not by hepatic lipase.


Cell culture studies of Menkes kinky hair disease. The dose response as well as kinetics of uptake and retention of copper and cadmium of normal and Menkes kinky hair disease (MKHD) cultured fibroblasts are described. In basal culture medium, intracellular copper concentration in MKHD fibroblasts was approximately 3 times that of control cultures. The intracellular copper concentration of MKHD cells was significantly higher than that of normal fibroblasts at medium copper concentrations below 20 microgram/ml. Death of MKHD cells occurred at medium copper concentrations between 15 and 20 microgram/ml with an intracellular copper level 3 times that at basal medium. Normal cells died at medium copper concentration above 30 microgram/ml with an intracellular copper concentration 19 times that at basal medium. These observations suggested the existence of a regulatory mechanism for maintenance and control of intracellular copper in normal fibroblasts which is effective at medium copper concentrations below 30 microgram/ml. This system is defective in MKHD fibroblasts. In basal medium MKHD and normal fibroblasts had similar intracellular cadmium concentrations; however, at higher medium cadmium concentrations MKHD cells had increased intracellular cadmium levels. The uptake of both 64Cu and 109Cd was significantly higher in MKHD cells than in normal cells, indicating that the uptake of 64Cu and 109Cd is not impaired in MKHD cells. A higher retention of 64Cu was observed in MKHD cells at both 37 degrees C and 4 degrees C. No obvious trend, however, was observed in the difference of retention of 109Cd between MKHD and normal cells. An impairment of egress of copper in MKHD cells is implicated by these results.


The incidence and likely origins of serum particulate alkaline phosphatase and lipoprotein-X in liver disease. Both serum particulate alkaline phosphatase and serum lipoprotein-X have been proposed as diagnostic markers for obstructive liver diseases. In this study their diagnostic efficiencies have been compared with other biochemical indicators of liver function and the relative incidence of these two markers has been determined in 241 patients with well defined liver disease. Although these markers appeared together most frequently in obstructive liver disease they were both present in some patients with liver disorders where obstruction was unlikely. One marker was present, independently of the other in 35% of the patients studied, mainly those without apparent cholestasis. The reasons for this are discussed in terms of the likely origins of the two markers and the probability that serum particulate alkaline phosphatase exists in two forms, as part of a multienzyme plasma membrane complex and as soluble liver enzyme associated with lipoprotein-X.


Reduced level of non-esterified fatty acids in sera from patients with infectious respiratory disease. Sera from 103 fasting individuals 3 to 76 years of age and free of clinical infectious disease and sera from 183 patients with infectious disease were assayed for serum total non-esterfied fatty acids (tNEFA) and compared. Data were also separated into five groups according to age of donor: 3--7, 8--19, 20--35, 36--60, and 61--76 years. The mean group serum levels of tNEFA increased with age. Among patients with infectious diseases sixty-five were diagnosed as having hepatitis, 41 with infectious mononucleosis, 18 with cellulitis, 12 with pulmonary tuberculosis, 11 with non-pneumococcal pneumonia, 9 with pneumococcal pneumonia, 8 with pharyngitis, 6 with pyelonephritis, 6 with aseptic meningitis, 4 with Gram-negative sepsis, and 3 with encephalitis. The sera from 23 non-fasting patients with gonorrhea were also tested. The serum tNEFA levels were found to be altered, in fact depressed from normal group values, only in patients with pneumonia or tuberculosis. This depression may be related to aberrant pulmonary metabolism during pneumonia.


A simple and rapid method for the estimation of serum haemoglobin binding capacity on Ultrogel AcA 44. A method is described for the estimation of serum haemoglobin binding capacity. The method is based on the separation of the high molecular weight haemoglobin . haptoglobin complex formed after addition of haemoglobin in excess to serum from uncomplexed haemoglobin by gel filtration through Ultrogel AcA 44. Agar gel electrophoresis and benzidine staining proved that the first fraction was exclusively the haemoglobin . haptoglobin complex. The second peak consisted of free haemoglobin. In this method the amount of haemoglobin bound to haptoglobin is calculated from the absorbance of the chromatographed complex at 412 nm. The results obtained are reproducible and comparable with those of an electrophoretic reference method, but less precise. The method is simple, and accurate, and is faster than gel filtration methods described earlier.


Plasma and urine amino acid changes in rats treated with hypoglycin. Hypoglycin A is a toxin which causes Jamaican vomiting sickness. In rats treated with this compound, we observed significant increases in a number of amino acids in the plasma and the urine, and detected several unidentified amino compounds in the urine. These changes suggest that hypoglycin, in addition to its known inhibitory effect on acyl-CoA dehydrogenases interferes with some steps in nitrogen metabolism as well. The amino acid abnormalities seen in hypoglycin-treated rats are compared with those seen in Reye's syndrome.


Improved and more specific bromcresol green methods for the manual and automatic determination of serum albumin. The bromcresol green (BCG) method for determination of serum albumin has been modified and the specificity improved. A manual and a continuous flow method is described. Both methods are suitable for routine use and the results agree well with those obtained by an immunonephelometric method.


A simple modification to an LKB 8600 reaction rate analyser for rapid measurement of enzymes showing lag phases in a coupled system. An enzyme assay which utilizes a coupled enzyme system may show a lag phase. To help to overcome problems associated with the kinetic measurement of these enzymes a modification to an LKB 8600 reaction rate analyser is described. It has been applied successfully to a kinetic method for the measurement of 5'-nucleotidase activity in serum.


Interference in estimation of serum copper concentration resulting from use of silicone-coated tubes for collection of blood. Prolonged contact between silicone-treated stoppers in evacuated tubes and serum was found to interfere in estimation of serum copper concentration using atomic absorption spectrophotometry.


Bromocriptine: lack of effect on the angiotensin II and aldosterone responses to sodium deprivation. Five normal subjects took a low sodium diet for four days on two occasions, one with and one without added bromocriptine 2.5 mg three times a day by mouth. Daily measurements of urinary electrolytes and the concentrations of plasma renin (PRC), angiotensin II (AII), aldosterone, 18-hydroxycorticosterone, cortisol, electrolytes and prolactin were made in both phases of the study. Deprivation of sodium without bromocriptine resulted in progressive and highly significant increases in the plasma concentration of aldosterone from 230 +/- 50 to 418 +/- 44 (SEM) pmol/l, 18-hydroxycorticosterone from 627 +/- 138 to 1420 +/- 478 pmol/l, PRC from 108 +/- 38 to 166 +/- 14 muU/ml and AII from 16 +/- 3 to 29 +/- 4 pmol/l. Similar changes were found during bromocriptine administration despite suppression of prolactin secretion. Sodium deprivation together with bromocriptine resulted in increases in the plasma concentrations of aldosterone from 230 +/- 47 to 416 +/- 72 pmol/l, 18-hydroxycorticosterone from 630 +/- 99 to 1629 +/- 552 pmol/l, PRC from 105 +/- 12 muU/ml and AII from 14 +/- 3 to 26 +/- 5 pmol/l. Plasma cortisol did not change either in response to sodium deprivation or bromocriptine. Mean cumulative negative sodium balance was 101 +/- 14 mmol on bromocriptine and 118 +/- 14 mmol in the control period. We conclude that prolactin is not necessary for the steroidogenic response to sodium deprivation in man.


Tissue growth hormone release inhibiting hormone-like immunoreactivity in experimental hypothyroidism and hypopituitarism. Hypothyroidism in rats was associated with an increase in immuno-reactive GH-RIH in brain, pancreas and gut, although release from the latter may be diminished as portal GH-RIH-like immunoreactivity was lower than control values. Hypophysectomy resulted in a depletion of immunoreactive GH-RIH in the septum and preoptic area of the brain and gastric antrum, but an increase in pancreas; portal venous GH-RIH-like immunoreactivity was not different from control concentrations, possibly reflecting both elevated and lowered immunol-reactive GH-RIH in different regions of tissue subserved by the portal vein. Inferior vena caval GH-RIH-like immunoreactivity was always lower than in the portal vein and was not influenced by tissue pertubations in hypothyroidism and hypopituitarism which made regional blood sampling of great important in evaluating tissue changes.


Hazards of amniocentesis: an unidentifiable fragment. A 38-year-old woman had amniocentesis performed for advanced maternal age. The karyotype of the cultured amniotic fluid cells showed a minute fragment in all cells analyzed, 47,XY+Fg. The fragment could not be identified. For medical reasons the pregnancy had to be terminated at 34 weeks. Both skin and blood cultures from the aborted fetus showed a 47,XY,21+ karyotype.


Ultrasonography for guidance of amniocentesis in genetic counseling. The value of preliminary ultrasonography as a guide for amniocentesis in the early second trimester of pregnancy was prospectively evaluated. One hundred and fifty patients were alternately assigned to a control group or to an ultrasound group who underwent examination with a real-time scanner immediately prior to amniocentesis. All amniotic fluid samples were assessed as to the presence of blood by: (1) visual observation during the amniocentesis; (2) appearance of the centrifugate; and (3) microscopic analysis. Ultrasonography did not reduce the failure rate, the incidence of multiple needle insertions, or the proportion of amniotic fluid samples containing blood.


Partial tetrasomy 9 in a liveborn infant. An unusual rearrangement of chromosome 9 was identified in a male infant with multiple congenital malformations. The rearrangement appeared as a fusion of two number 9 chromosomes with similar long-arm breakpoints. Since the infant also possessed two normal 9's, the presence of the additional chromosome resulted in partial tetrasomy; 47,XY, + tdic(9;9)(q22;q22). Clinical and autopsy examinations revealed many features reminiscent of trisomy 13. The tdic was functionally monocentric, although some evidence of activity at the second centromere was observed. Both parents had normal karyotypes, and C-banding demonstrated that at least one of the 9h regions on the tdic was likely to be of maternal origin.


Dominant inheritance of multiple epiphyseal dysplasia, myopia and deafness. An Afrikaner kindred in South Africa had a dominantly inherited skeletal dysplasia-blindness-deafness syndrome. The affected individuals had reduced stature and a round, flattened facies. The bone changes resembled multiple epiphyseal dysplasia with additional minor abnormalities in the phalanges, femoral heads and spine. Progressive myopia, retinal thinning and crenated cataracts led to visual disturbance, while conductive deafness represented the third component of the triad. The syndromic significance of associated asteroid hyalosis is uncertain.


The cloverleaf skull.

The cloverleaf skull. A patient presented with the cloverleaf skull, or Kleeblattschädel abnormality. He had a typical trilobed skull with synostosis and a honeycomb appearance on roentgenograms. He had no extracranial skeletal anomalies. The infant died at 1 month of age.


Partial trisomy of the short arm of chromosome 7 due to a familial translocation rcp(7;14)(p11;p11). A boy with trisomy of the short arm of chromosome 7 is reported. He presented several congenital malformations, such as: dolicocephaly; wide fontanelle; hypertelorism; antimongoloid slanting of the palpebral fissures; small, carp-shaped mouth; thin neck; extrinsic compression of the oesophagus; congenital dislocation of the shoulders and hips; and a possible cardiac defect. The proband's mother had a balanced translocation involving the short arm of chromosomes 7 and 14. In this family the translocation has been transmitted through three generations.


The Lp lipoprotein in Japanese.

The Lp lipoprotein in Japanese. The Lp gene frequency is estimated to be 0.083 in Japanese, but with our method of assay the evidence favoring a major locus is marginal. The sinking pre-beta band is significantly associated with elevated cholesterol and depressed triglyceride. The biological significance of these observations is discussed.


Low arylsulphatase A activity in a family without metachromatic leukodystrophy. A low arylsulphatase A activity was noted in the leukocytes and cultured skin fibroblasts of a child without any other symptoms of metachromatic leukodystrophy. Although the mother had a level of arylsulphatase  commensurate with heterozygosity for the classical metachromatic leukodystrophy gene, the father had a variant gene giving an unusually low in vitro level of this enzyme. In combination (the proband), these two genes gave rise to a very low in vitro activity without any apparent disease. In screening for metachromatic leukodystrophy, a low arylsulphatase A level is not necessarily indicative of this disease, if a clinically normal parent shows an unusually low level of this enzyme.


Familial ovarian carcinoma: a problem in genetic counselling. Ovarian papillary adenocarcinoma is reported in two families. In one family, this tumor was detected in four females, of whom two had had mammary cancer below the development of the ovarian tumor. In the second family, a mother and daughter died of ovarian adenocarcinoma. The mode of inheritance in both families fits the pattern of autosomal dominant transmission of a single gene. Both are Jewish families from Germany, presently residing in Israel. The two families are presented as examples illustrating the difficult question of genetic counselling in families with recurrent cases of ovarian carcinoma. The difficulty arises because this tumor is relatively frequent but is usually observed in isolated cases without systematic follow-up in the patients' families. Only a few cases, where more than one person in a family is affected, have been selectively reported in the literature.


Partial trisomy 15 and temporal lobe syndrome in a retarded girl without gross malformations. A supernumerary small bisatellited chromosome was found in a girl with stunted growth and psychomotor retardation. The extra chromosome was identified as a deleted 15, del(15)(q21), with C-band positive heterochromatin and satellite-like appendages to the distal end of the long arm. This chromosome was the product of a translocation between a chromosome 15 and some other acrocentric chromosome, as shown by G-, C- and Q-banding and silver staining of the nucleolus organizer regions. The proposita had no gross phenotypic malformations. She had a small head, a high forehead, oblique palpebral fissures, bilateral enophthalmus, clinodactyly and simple dermatoglyphic patterns. She was autistic and suffered from epileptic seizures and expressive aphasia. The waking electroencephalogram revealed diffuse abnormalities; sleep recording showed focal spikes and sharp waves anteriorly on the left side. The pneumo-encephalogram showed microventriculy, an enlarged left temporal horn and some enlarged sulci in the right frontotemporal cortex. The prenatal influence of the chromosome anomaly is interpreted as being the primary cause of these disorders, neonatal asphyxia being a secondary contributing factor.


An XYY boy with short stature and a case of Klinefelter's syndrome (XXY) in a family with inversion 9. A patient with Klinefelter's syndrome and a boy with XYY sex chromosomes were both found to have a pericentric inversion of chromosome 9. An unusual feature of the XYY patient was that he presented because of short stature and disturbed behaviour. A family study showed that the patients were related and that there was an excess of males in the pedigree. Another member of the family was found to have some XYY cells in the blood.


Peripheral muscle effects of levodopa in the anesthetized cat. 1. Levodopa, dopamine, noraderanaline and adrenaline (in increasing order of potency) depressed the tension and degree of fusion of incomplete tetanic contractions of the slow-contracting soleus muscle in chloralose-anaesthetized cats. 2. The effects of all compounds were antagonized by propranolol (50-20 microgram/kg), but not practolol (1.0-5.0 mg/kg). This indicates that effects are mediated by beta2-adrenoceptor stimulation. 3. The effect of levodopa, but not of the catecholamines, was antagonized by prior administration of the dopa decarboxylase inhibitior benserazide. This indicates that levodopa itself is inactive, whereas  its decarbodylated metabolites are active. 4. The depressant action of beta-adrenoceptor agonists on incomplete tetanic contractions of the cat soleus muscle, which are exerted directly on the muscle fibres, is a model for effects exerted on slow-contracting units of human muscles; the latter effects probably underlie the tremor observed after beta-adrenoceptor agonist administration. 5. These results therefore suggest that levodopa, via its decarboxylated metabolites, dopamine, noradrenaline and adrenaline, may produce or exacerabate tremor in man. Thus in Parkinsonian patients any centrally induced relief of tremor that levodopa may produce may be masked by tremorogenic effects of such metabolites exerted in the periphery.