Prospective evaluation of periodic breast examination programs. A mathematical model is developed to obtain prospective estimates of average tumor sizes and the expected proportion of positive regional lymph node cases for periodic breast cancer examination programs. Semiannual, annual, and biannual programs, with or without mammography, are evaluated and compared with results from the National Breast Poject. Calculations indicate that a semiannual clinical-mammographic program can be expected to result in about 37% fewer positive node cases than those observed in the National Breast Project and substantially small tumors. The expected reduction in positive node cases from a similar annual program is about 30%, and from a biannual program, even without mammography, about 20%. Calculations also indicate the important role of diligent patient self-examination in such programs, especially for the detection of fast-growing tumors.


Effect of hyperthermia on the radiosensitivity of normal and malignant cells in mice. The effect of systemic hyperthermia on the in vivo radiation response of normal and malignant mouse cells was evaluated. X-irradiation of L1210 cells and Ehrlich ascites cells at body temperatures above 41 degrees C resulted in strongly enhanced tumor cell death. The magnitude of this thermal effect increased with increasing temperatures. Hypoxic tumor cells were particularly sensitive to combined heat-radiation treatment. L1210 leukemia cells did not become resistant to the sensitizing effects of hyperthermia even after repeated heat exposures over several transplant generations.  The sensitizing action of hyperthermia varied with different heating strategies. Heating before or during irradiation did not materially alter the radiation response of tumor cells. Maximal potentiation of radiation damage was achieved only when the tumorous mice were subjected to at least 20 minutes heat incubation after irradiation. LD studies on ICR mice revealed that moderate hyperthermia (41.5 degrees C) does not alter  the radiation response of normal body tissues. These findings indicate that it is possible to devise hyperthermic treatment regimens that drastically enhance radiation-induced tumor cell death in vivo without reducing the radioresistance of normal tissues.


Survival and complications of radiotherapy following involved and extended field therapy of Hodgkin's disease, stages I and II. A collaborative study. A collaborative trial with random assignment to involved field or extended field radiotherapy for localized Hodgkin's disease was begun in 1967. Case accession was completed in 1973, with 224 eligible patients assigned to involved field and 243 to extended field. With a median follow-up time of 27 months no significant survival difference is found between involved and extended field regimens for the total patient group or for most subgroups defined by age, sex, histology, stage, class, and use of laparotomy for staging. A single exception is an improved survival in the involved field group for female patients. Survival free of distant extension shows a similar lack of treatment effect, but survival free of any extension, local or distant, shows a significant benefit from extended field treatment. Complications of radiotherapy are significantly increased following extended field treatment. Survival following local extension is similar to survival following periods free of any extension.


Fibroelastic hamartoma (fibroma) of the heart. Approximately 70 cardiac fibromas (fibroelastic hamartomas) have been reported in the literature and at least 15 have been successfully excised. However there is no well-documented ultrastructural study of these lesions. A successfully excised cardiac fibroma (FEH) was studied by light and elctron microscopy. The tumor was composed mainly of fibroblasts admixed with bundles of collagen and elastic fibers. No muscle fibers were demonstrated in the central region of the tumor either by light or electron microscopy. Mast cells were occasionally present. The lack of encapsulation enabled fibroblasts with collagen and elastic fibers to extend between groups of myocardial fibers at the periphery of the tumor. Within these myocardial fibers ultrastructural changes were limited to the mitochondria and myofibrillar structure.


Morbidity and mortality among recipients of blood from preleukemic and prelymphomatous donors. The present investigation was designed to test the admittedly speculative hypothesis that a factor involved in the etiology of human lymphatic and hematopoietic neoplasms may be transmitted by blood transfusion prior to the clinical onset of illness in the donor. One hundred and five New York State residents, who received blood from donors who subsequently developed neoplasms of the lymphatic or hematopoietic tissues, were identified and followed for an average period of 7.05 years. No recipient was found to have developed a leukemia or lymphoma following receipt of blood from a preleukemic or prelymphomatous donor. The results of this study should be considered priliminary because the small size of the recipient group might mask even a large increase in risk among the recipients.


Modes of invasion of cancer of the larynx. A statistical, histological, and radioclinical analysis of 120 cases. A statistical comparison between histologic and radioclinical data was performed on a series of 120 cases of total laryngectomies in order to study the modes of invasion of laryngeal cancer and to quantify the differences of results as evaluated by pathological and by clinical examinations. The first part of this study deals with the spread of cancer depending on its site of origin; the second part examines spread regardless of the primary location of the laryngeal tumor. The comparative results show that it was difficult to determine the deep invasion of structures such as the preepiglottic space and the cartilages from the radioclinical examination, especially in cases of cancer developed on a laryngocele. This study emphasizes the importance of the histological examination of laryngeal specimens as described here in order to codify the modes of invasion and, as a result, to help the otorhinolarynglologist in the choice of the best treatment.


Ectopic thyroid tissue in the neck. Benign or malignant? This is a report of ectopic thyroid tissue in the neck, associated with a nodular colloid goiter, which recurred at least three times, beginning at age 24 years, in a woman in 12 years. The ectopic tissue appeared histologically benign and was identical to that found in the thyroid gland. Scintiscans of the neck and thyroid suppression tests showed that the tissue was initially unsuppressible and presumably autonomous in its function. Our conclusion is that the most reasonable explanation for this phenomenon is the intraoperative transmission of thyroid cells, probably benign and autonomous in function, to other sites in the neck.


Medullary carcinoma of the thyroid. Diagnostic problems. Light microscopy is usually considered sufficient for the diagnosis of medullary carcinoma of the thyroid (MCT). As stromal amyloid is not always present and the tumor may exhibit great variation in growth pattern, light microscopy alone, however, may lead to misinterpretations. Of 1670 thyroid carcinomas registered durging a 15-year period in Norway, 42 were originally interpreted as MCT. The slides were reviewed and the diagnosis maintained in 33 cases only. Twenty-four additional cases were found by reviewing histopathologic slides from neoplasms originally registered as other types of thyroid tumors. Of 57 cases of MCT, stromal amyloid was demonstrated in 43. Calcitonin measurements and electron microscopy, even on formalin-fixed material, were valuable aids in establishing the correct diagnosis, though none of these methods are unequivocal. Different aspects of the problems concerning the diagnosis of MCT are discussed through the detailed presentation of five patients.


Abdominoperineal resection following anterior resection. A series of 11 patients undergoing abdoninoperineal resection for "suture line recurrence" following anterior resection is presented. Five-year survival is 10%. Technically, the procedure is difficult and major problems are encountered, including large blood loss and ureteral complications. These patients had an inadequate distal margin of resection at the time of anterior resection. The survival of this group of patients underscores the importance of making the correct judgment about anterior or abdominoperineal resection at the time of the initial presentation of the patient. The phrase "suture line recurrence" is a misnomer; all of these patients had advanced pelvic malignancy. If the adequacy of the distal margin is questionable or a distal margin of 5 cm cannot be obtained safely at the time of anterior resection, abdominoperineal resection should be performed, as the opportunity for cure of a recurrence should this rule be compromised is limited.


College athletics, body size, and cancer mortality. Data are presented on mortality from neoplasms as determined from death certificates in a cohort of 8393 college men, according to athletic status in college. Major athletes (lettermen) died significantly more often from neoplasms than nonathletes. Mean age at death from neoplasms (underlying cause) was significantly lower in major athletes than in both minor athletes and nonathletes. After matching major athletes with nonathletes of comparable body size (height and weight), differences in proportional mortality and mean age at death from neoplasms persisted, although not statistically significant for the smaller samples. Correlation coefficients (Pearson r) and partial r's between weight in college and age at death from neoplasms were negative but of low magnitude. Some possible explanations for the differences between major athletes and nonathletes are discussed.


The effect of measuring error on the results of therapeutic trials in advanced cancer. In this study, 16 experienced oncologists each measured 12 simulated tumor masses employing their usual clinical methods. Unknown to the oncologists, two pairs of these tumors were identical in size. This permitted a total of 64 measurement comparisons of the same investigator measuring the same size mass and 1920 comparisons of different investigators measuring the same size mass. If a 50% reduction in the product of perpendicular diameters is accepted as a criterion, the objective response rate due to measuring error alone was 7.8% by the same investigator and 6.8% by different investigators. If a 25% reduction criterion is used, the respective "placebo" response rates were 19% and 25%. In the clinical setting it is recommended that the 50% reduction criterion be employed and that the investigator should anticipate an objective response rate of 5 to 10% due to human error in tumor measurement.


Combinaed chemotherapy with cyclophosphamide, vinblastine, procarbazine, and prednisone (CVPP) for patients with advanced Hodgkin's disease. An alternative program to MOPP. Thirty-eight patients with advanced Hodgkin's disease were treated with a combination of cyclophosphamide, vinblastine, procarbazine, and prednisone (CVPP). Complete remissions were obtained in 74% of patients. Response rates were similar for previously untreated and previously extensively irradiated patients. Fifty-six percent of patients older than 50 and 50% of patients with bone marrow involvement responded. All patients with mixed cellularity Hodgkin's disease responded compared with 74% of patients with nodular sclerosis. At the  present time the median duration of complete remission is 27+ months. However, of the responders, 75% (21 of 28) remain in continous complete remission for 14+ to 49+ months. Only five patients have relapsed but two have died from apparent sepsis without evidence of recurrent disease. The complete remission rate for CVPP is comparable of that for MOPP, CVPP is an easily administered, well-tolerated alternative to MOPP chemotherapy for patients with advanced Hodgkin's disease.


Further clinical studies with megestrol acetate in advanced breast cancer. A previous study showed activity of megestrol acetate in advanced breast cancer; as a result the study was expanded. Of 101 patients treated with this compound, 26 (26%) met our criteria of improvement. The drug was well tolerated and produced no toxicity and, as opposed to androgens and estrogens, no endocrine effects were observed. Prior hormonal or cytotoxic compounds, including alkylating agents, did not appear to reduce the subsequent responsiveness to this potent progestational compound. We now include it, often as the initial hormonal trial in postmenopausal patients, in our sequential treatment of disseminated breast cancer because of its favorable therapeutic ratio.


Repeat lymphography in children with Hodgkin's disease. Repeat lymphography was performed in 37 children less than 16 years  old with histologically proven Hodgkin's disease. The repeat studies had the same lymphatic cannulation success rates as the first lymphogram; one patient had a bilaterally unsuccessful repeat study, which left 36 for analysis. Both first and repeat lymphograms were negative in 33% of children, whereas a negative lymphogram became positive for tumor in 39%. The remaining 28% had a positive first lymphogram, with an equal incidence of positive and negative repeat studies. Reactive hyperplasia was seen in 16.7%, and post-therapeutic lymph node alteration in 22%, of all cases. Of the 22 children studied for routine opacification or because of suspected but unproved relapse, 10 (45%) had positve repeat studies. The repeat lymphogram in these patients was of utmost importance in initiating therapy. Nine of 14 patients studied for documented clinical relapse had positive repeat studies.


Relation of certain infrared bands to conformational changes of cellulose and cellulose oligosaccharides. The i.r. spectra of D-glucose and cellulose oligosaccharides up to cellopentaose have been compared with those of cellulose at various temperatures between that of liguid nitrogen and approximately 250 degrees. Significant changes in frequency and intensity of the bands at approximately 3400 cm -1 were observed. The a 1372 cm -1/a2900 cm -1 ratio for each carbohydrate studied decreased gradually as the temperature was increased above ambient. The change of the band intensities at 1429 and 893 cm -1 with temperature was also investigated. The observed spectral changes are assumed to be associated with changes of hydrogen bonding.


An endo-(1 lead to 6) -beta-D-glucanase from Mucor hiemalis. An endo-(1 lead to 6)-beta-D-glucanase (EC 3.2.1), isolated from the culture filtrate of Mucor hiemalis, was purified by ammonium sulphate fractionation and gel filtration. The homogeneity of the enzyme was confirmed by disc electrophoresis. The enzyme had a wide range of temperature and pH stability, high substrate specificity, and an action pattern of the endo-type.


Methylation and acetolysis of extracellular D-mannans from yeast. Methylation-fragmentation analyses were conducted on a series of extra-cellular, yeast alpha-D-linked mannans representing six different structural types. D-Mannans of low degree of branching were produced by Hansenula capsulata strains and by species related to H. holstii, The former consisted primarily of (1 leads to 2)- and (1 leads to 6)-linked D-mannosyl residues; the latter, of (1 leads to 2)- and (1 leads to 3)-linked D-mannosyl residues. Although the remaining structural types were highly branched, each gave distinct methylation-patterns indicative of (1 leads to 6)-linked backbones to which are appended non-(1 leads to 6)-linked side-chains. Acetolysis studies were correlated with the methylation analyses, and the correlation demonstrated that each branched polymer possesses side chains of heterogeneous length.


alpha-D-Glucopyranosyl fluoride as a D-glucopyranosyl donor for a glycosyltransferase complex from Streptococcus mutans FA1. alpha-D-glucopyranosyl fluoride can serve as the D-glucopyranosyl donor for the glycosyltransferase from Streptococcus mutans. The products of the reaction are a D-glucan of high molecular weight and fluoride ion. The rate of reaction was measured by an electrode specific for the fluoride ion. The reaction was inhibited by the substrate at concentrations greater than 30 mM, but was not affected by fluoride ion. There was little inhibition of the reaction by a series of monosaccharides, except for 2-amino-2-deoxy-D-glucose, D-mannitol, and 3-deoxy-D-ribo-hexose. Maltose, isomaltose, isomaltotriose, and D-fructose all stimulated the release of fluoride from alpha-D-glucopyranosyl fluoride.


Purification of malted-barley endo-beta-D-glucanases by ion-exchange chromatography: some properties of an endo-barley-beta-D-glucanase. Two endo-beta-D-glucanases which act, respectively, on (1 leads to 3)-beta-D-glucans and barley beta-D-glucan have been isolated from malted barley, and purified by ion-exchange chromatography. The latter enzyme is highly specific for barley beta-D-glucan, and has no action on either (1 leads to 3)- or (1 leads to 4)-beta-D-glucans. It will also act on dyed barley-beta-D-glucan. Certain group-specific reagents inhibit the endo-barley-beta-D-glucanase and the endo-(1 leads to 3)-beta-D-glucanase to similar extents.


Absence of messenger RNA and gene DNA for beta-globin chains in hereditary persistence of fetal hemoglobin. The relative amounts of alpha-amd beta-globin mRNA and globin gene DNA were measured in reticulocyte RNA and lymphocyte DNA of an individual with homozygous hereditary persistence of fetal hemoglobin whose red blood cells contain 100% fetal hemoglobin (hb F: alpha2gamma2.) Molecular hybridization assays used as probes full-length DNA copies of human alpha- and beta-globin messenger RNA. The results of these hybridization assays demonstrated the expected amounts of alpha-globin mRNA and gene DNA, but absence of beta-globin mRNA and absence of beta-globin gene DNA. In the individual studied, hereditary persistence of fetal hemoglobin is associated with total deletion of the beta-globin structural gene.


The blocked and methylated 5' terminal sequence of a specific cellular messenger: the mRNA for silk fibroin of Bombyx mori. The messenger RNA for silf fibroin, laveled with 32PO4 and methyl-3H L-methionine, was purified to near homogeneity from the posterior silk gland of the sildworm Bombyx mori, and the sequence of a methylated, RNAase T2-resistant structure was determined. This sequence is similar structurally to 5' terminal blocked and methylated sequences found on the total populations of polyadenylated eucaryotic cellular and certain viral mRNAs. The RNAase T2-resistant oligomer from fibroin mRNA was cleaved by nucleases P1 into three components: a blocked and methylated sequence containing three phosphates; a 2'-0-methyl UMP residue (pUm), and an unmethylated CMP (pC). The blocked and methylated sequence comigrated in three chromatographic systems with the blocked and methylated terminus of silkworm cytoplasmic polyhedrosis virus mRNA, which has the structure m7GpppAm. The fibroin mRNA cap was cleaved by nucleotide pyrophosphatase to yeild 7-methyl GMP Tpm7G) and 2'-0-methyl AMP (pAm). This sequence also appeared to be terminally located, with the m7G joined by a 5'-5' pyrophosphate linkage to the Am. It was concluded that the 5' terminal sequence of fibroin mRNA molecules is m7G(5')ppp(5')AmpUmpCp. The regulation of expression of the highly specialized gene for fibroin is discussed in light of this finding.


Yeast suppressors of UAA and UAG nonsense codons work efficiently in vitro via tRNA. A cell-free protein-synthesizing system, containing an S-100 fraction from yeast, ribosomal subunits from Krebs ascites cells, and ribosome initiation factors from rabbit reticulocytes, translates yeast, adenovirus, and rabbit globin messenger RNAs and the RNA from bacteriophage Qbeta. An amber mutation in the Qbeta synthetase gene is suppressed in vitro if the S-100 fraction s from yeast strains carrying amber suppressor mutations. Suppressor SUP6-2 gives 16% suppression, and the recessive lethal suppressor RL-1 gives 50% suppression. Extracts from strain FM6, which has the ochre suppressor SUP4-1, give a longer protein product from the normal synthetase gene at Qbeta with an efficiency of 63%. This implies that UAA is the terminator for the synthetase gene, and that synthesis of this read through protein can be used as an assay for ochre suppression. Suppression in each of these cases is mediated by tRNA, since pufified tRNA is the only fraction from suppressing strains that is required in an otherwise nonsuppressing cell-free system.


Elevated dihydrofolate reductase messenger RNA levels in methotrexate-resistant BHK cells. Polysomal RNA from cultured sublines of baby hamster kidney (BHK) cells directed protein synthesis in an in vitro system derived from wheat germ extract. One product of the in vitro synthesis was dihydrofolate reductase (DHFR), as confirmed by methotrexate-substituted Sepharose affinity chromatography followed by SDS-polyacrylamide slab gel electrophoresis and autoradiography of the proteins labeled with 35S-methionine. The DHFR synthesized in vitro comigrates in the gel with authentic BHK DHFR, indicating that the molecular weights and structures  of the in vivo and in vitro enzymes are probably the same. Polysomal RNA obtained from the methotrexate-resistant BHK subline (A5), which possesses some 140 times higher DHFR levels than the methotrexate-sensitive parents subline (B1), directed the synthesis of approximately 70 times more DHFR per unit of total in vitro synthesized protein than did B1 polysomal RNA. Assuming then that the rates of translation of A5 and B1 DHFR mRNAs in the wheat germ cell-free system are the same, our results show that a major part of the high DHFR levels observed in A5 cells is due to the presence of elevated quantities of DHFR mRNA.


Effects of protease treatment on growth, morphology, adhesion, and cell surface proteins of secondary chick embryo fibroblasts. Several proteolytic enzymes have been studied with regard to their ability to induce DNA synthesis and cell proliferation in resting chick embryo fibroblasts. Of the enzymes examined, thrombin, bromelin, and trypsin exhibit potent mitogenic activity, elastase has significant but less marked activity, whereas thermolysin, papain, and alpha-protease are inactive. The enzymes were also tested for their ability to induce morphological change or to remove two iodinatable proteins of 250,000 and 205,000 daltons. Although the larger protein is removed by some but not all of the proteases examined, every protease tested removed the smaller cell surface proteins; however, loss of the smaller protein does correlate with the reduction of both cytoplasmic spreading and cell-cell interactions observed after protease treatment. A secondary, later event of migration of cells into clumps is observed in those instances when protease treatment did not result in a loss of the 250k protein. Arole for each of these proteins in the processes of cellular adhesion is discussed.


Feminization of hepatic S metabolism in male rats with a transplanted (MtT/F4). After transplantation of MtT/F4 pituitary tumor cells to male rats of the Fisher strain, the masculine type of hepatic steroid metabolism was changed into a feminine pattern of enzyme activities. Liver metabolism of steroid hormones in female rats was relatively unaffected following transplantation of pituitary tumor cells. Futhermore, extract from MtT/F4 tumors and "autonomous" pituitary tissue increased the 5alpha-reductase activity of hepatoma cells in the culture (HTC cells) at subsaturation concentrations of the substrate 4-androstene-3, 17-dione by decreasing the apparent Km of the enzyme. It is concluded that the pituitary tumor (in accord with the secretion from an "autonomous" pituitary gland) secretes "feminotropin," a novel hypophyseal principle that probably is an important regulator of hepatic steriod metabolism. It is suggested that pituitary tumor tissue of the MtT/F4 type could be used as source of feminotropin in purification studies.


A unique pattern of integrated viral genes in hamster cells transformed by highly oncogenic human adenovirus 12. We have analyzed the viral DNA sequences persisting in HE C19, a clonal line of hamster embryo cells transformed in vitro by human adenovirus 12 (Ad 12), a member of highly oncogenic group A. Saturation-hybridization of in vitro labeled Ad 12 DNA (2-4 x 10(8) cpm/mug) with HE C19 DNA showed that 93-100% of viral DNA sequences are present in the transformed cell. Reassociation of Ad 12 32P-DNA in the presence of HE C19 DNA detected 8 viral genome equivalents per diploid quantity of cell DNA. REannealing each of the six Ad 12 EcoRl restriction DNA fragments with HE C19 DNA indicated that the sequences of each fragment are present in near equimolar quantities, 8-10.5 copies per cell. HE C19 DNA "networks" were prepared and shown to contain the same viral DNA sequences and in the same amounts as present in unfractionated transformed cell DNA; these data provide the first direct evidence that human adenovirus genes are covalently linked to cellular DNA. The presence of most, if not all, of the viral genes was demonstrated also in the Ad 12 hamster tumor cell line, HT-2. The pattern of integrated viral genes in these nonpermissive hamster cells transformed by Ad 12 is different from that described for cells transformed by group C and B human adenoviruses and for SV40.


Amino acid starvation affects the initiation frequency of nucleolar RNA polymerase. The synthesis of ribosomal precursor RNA in mouse ascites nucleoli derived from cells starved for amino acids is compared with the activity of nucleoli from control cells cultivated in the presence of all amino acids. It is shown that deprivation of a single essential amino acid from the culture medium results in a drastic decrease of the RNA-forming capacity of the isolated nucleoli by a factor of 2-3. This switchoff in rRNA synthesis is a very fast process. Half-maximal inactivation occurs after only 30 min. Addition of amino acids to starved cells leads to a rapid recovery, which is reflected by a sharp increase in the RNA polymerase activity of the isolated nucleoli. Studies on the molecular mechanism of this amino acid-mediated control of rRNA synthesis indicate that this effect is not caused by different growth rates of the RNA chains, but rather by an altered initiation frequency of the RNA polymerase in vivo. Whereas in nucleoli derived from cells grown in full medium almost all the polymerase is tightly bound in a transcriptional complex, a high amount of "free" polymerase which becomes active after addition of exogenous template is present in nucleoli from starved cells.


Control of nucleolar RNA synthesis by the intracellular pool sizes of ATP and GTP. The influence of amino acid starvation on both the pool sizes of nucleoside triphosphates and the rRNA synthetic capacity of Ehrlich ascites cells was studied. The results indicate that under shiftdown conditions, an immediate shrinkage of the cellular ATP and GTP levels occurs. Concomitant with this, protein and rRNA syntheisis are markedly inhibited. If the pool sizes of purine nucleaside triphosphates are expanded by adding adenosine or guanosine to cells cultured in histidine-free medium, the nucleolar RNA synthesis is fully restored, while protein synthesis remains inhibited. The results suggest that the rate of pre-rRNA transcription may be controlled by the actual nucleoside triphosphate levels of the cells rather than by short-lived protein(s), as has been previously postulated.


Platelet aggregation in partially obstructed vessels and its elimination with aspirin. In 35 open chest anesthetized dogs coronary and aortic blood flow were measured with electromagnetic flowmeters while aortic and distal coronary blood pressure and an epicardial ECG were recorded. A fixed amount of stenosis (60-80%) was produced in the coronary artery by an externally applied plastic cylinder. In 24 of the 35 dogs the coronary blood flow showed cyclical reductions to near zero, with a sudden spontaneous return to near control levels. During reduced flow the epicardial ECG showed ST-segment depression suggestive of ischemia, and ventricular premature beats were often noted. Six animals died acutely during episodes of reduced flow. After 35 mg/kg of aspirin were given intravenously the cyclical reductions in coronary blood flow were abolished and the in vitro platelet aggregations were reduced from a control of 62.1 +/- 15 units (Born technique) to an average of 23.7 +/- 12 units. Histologic sections of the narrowed coronary artery obtained when coronary flow was reduced show an amorphous mass in the lumen which was thought to be a platelet aggregate. Perhaps a similar process of platelet aggregation occurs in the stenosed coronary arteries in man, producing acute coronary obstruction, ischemia, and sudden death.


False positive diagnostic tests and coronary angiographic findings in 105 presumably healthy males. Among 2014 presumably healthy males aged 40-59 years coronary heart disease (CHD) was suggested in 115 in the presence of one or more of the following criteria: 1) a WHO-questionnaire on angina pectoris positive on interview, 2) typical angina during a near maximal bicycle exercise test, 3) a positive exercise ECG during and/or post exercise, 4) a Minnesota Code 1.1 on a resting ECG. Diagnostic coronary angiography was offered to all 115 CHD-suspect cases. Six refused angiography and four others were excluded. Of the remaining 105, thirty-six had less than 50% obstruction of any major coronary artery (34.3%). Eighteen (17.1%) had single, 25 (23.8%) had double and 26 (24.8%) had triple vessel disease. In 62 of the 69 with pathologic angiograms at least one obstruction greater than or equal to 75% was found. Eighty percent of the cases with proven CHD were greater than or equal to 50 years. All CHD-suggestive criteria had approximately the same diagnostic performance regardless of age, i.e., approximately one false positive/two true positives. Except for one retroperitoneal hematoma no complications to angiography occurred.


Differences in muscle blood flow in upper and lower extremities of patients after correction of coarctation of the aorta. Using the method 133Xe clearance we investigated blood flow and calculated vascular resistances simultaneously in the muscles of the upper and lower extremities in 58 patients following successful surgical correction of aortic coarctation carried out at age 11.5 (+/- 2.9) years. The interval from operation to investigation was 11.5 (+/- 4.5) years. Resting and maximal ischemic exercise blood flows in the upper extremity were decreased and the duration of maximal blood flow was shortened. Values recorded from the lower extremities did not differ from normal controls. The difference between upper and lower extremities was statistically significant. Vascular resistance during maximal blood flow was higher in the upper extremities than in the lower. Differences between upper and lower extremities did not change after vasodilation elicited by amyl nitrite. The degree of differences was not dependent upon the age at operation, the age of the patients at investigation, or on the time interval between operation and investigation.


Pathogenesis of paradoxical hypertension after coarctation resection. The pathogenesis of paradoxical hypertension after resection of coarctation of the aorta was investigated by comparing the course of seven children undergoing repair of coarctation with five acyanotic children undergoing elective cardiovascular surgery. During the first 24 hours after surgery, all coarctation patients demonstrated a rise in systolic blood pressure (35 +/- 15.5 mm Hg; P less than 0.001), a significant depression in cold pressor test response, and only a slight elevation in plasma renin activity. In the next 24-72 hours, coarctation patients developed a rise in diastolic blood pressure (26.8 +/- 10.6 mm Hg; P less than 0.001), plasma renin activity (22.9 +/- 10.2/ml/hr; P less than 0.001) and fluid retention. By contrast, control patients had no significant postoperative changes. Abdominal pain occurred in five coarctation patients during the period of maximal plasma renin activity. The data suggest that the sympathetic nervous system may be responsible for the initial phase of hypertension after coarctation resection and that the renin angiotension system plays a major role in the second phase of hypertension and in the pathogenesis of mesenteric arteritis.


Quantitative measurement of left ventricular volumes in man from radiopaque epicardial markers. The distances between three or four radiopaque markers located on the left ventricular epicardial surface at the apex and in the region of the minor axes in 22 subjects showed close correlations with left ventricular chamber radii, length, volume and wall thickness determined from biplane angiograms over the range of stroke volume. The markers were previously placed during heart surgery. Regression equations relating the distances between epicardial markers and chamber volumes were used to predict volumes for other beats. There was close agreement of end-diastolic, end-systolic, and stroke volumes as determined by the angiographic and epicardial marker methods for premature atrial contraction beats and post premature contraction beats, and in studies performed during rest and exercise. Time-volume curves determined by the epicardial marker and angiographic methods were similar. It is concluded that epicardial marker measurements can be used to quantitate beat-to-beat left ventricular chamber volume and dimension changes.


Potential role of QT interval prolongation in sudden infant death syndrome. To investigate the possibility that a genetically transmitted cardiac abnormality is involved in the genesis of the sudden infant death syndrome (SIDS), 42 sets of parents who had at least one infant with SIDS were studied by electrocardiography. Prolongation of the QT interval was present in at least one member of 11 (26%) sets of parents. In families in which QT interval prolongation was found in a parent, prolonged QT interval was also present in 39% of the siblings of infants with SIDS, suggesting an autosomal dominant pattern of inheritance. In addition, an infant with "near-miss" SIDS showed marked prolongation of the QT interval. Thus, our data suggest that prolonged QT interval may play a role in a considerable proportion of sudden and unexpected infant deaths. However, definitive confirmation of the relation between QT interval prolongation and SIDS will require large prospective investigations.


Analysis of secondary pauses following termination of rapid atrial pacing in man. The first ten cycles following cessation of atrial pacing were evaluated in 44 control subjects (mean age 52.9 +/- 14.88 yr) and 39 patients (mean age 62.9 +/- 15.41 yr) suspected of having sinus node dysfunction (SND). The maximal cycle length for each postpacing cycle following several pacing periods in each control subject was determined, and was normalized by dividing it by the subject's mean spontaneous control cycle length (SCL). Using the control group, a normalized maximal post-pacing response pattern (mean and SD) was derived. For each SND patient, a composite SD for each post-pacing cycle was calculated by adding the patient's SCL variance to the variance determined for each post-pacing cycle in the control group. Two composite SD above the mean value for each post-pacing cycle was selected as the upper limit of the normal recovery response and used to identify abnormal post-pacing responses in the 39 SND patients. Abnormally prolonged cycle lengths subsequent to the first escape cycle (secondary pauses) were found in 16/39 (41.0%) patients, of whom 11/39 (28.2%) had a prolonged SNRTmax. Of importance, 11/12 (91;7%) patients with documented SA block or sinus pauses prior to electrophysiologic study, demonstrated secondary pauses, while only 7/12 (58.3%) had a prolonged SNRTmax. Criteria are derived for the identification of secondary pauses during the postpacing period, and a close association between secondary pauses and the presence of spontaneous SA block or sinus pauses prior to electrophysiologic study is demonstrated.


Electrophysiologic effects of propranolol on sinus node function in patients with sinus node dysfunction. The electrophysiologic effects of intravenously administered propranolol (0.1 mg/kg) on three parameters of sinus node function were examined in ten symptomatic patients with sinus node dysfunction. The patients ranged in age from 26 to 79 years. Symptoms ranged from fatigue to frank syncope. Sinoatrial (SA) block and sinus pauses were observed in one patient; sinus pauses alone were observed in three patients. Five (5/10) patients had intraatrial block; three (3/10) patients had atrioventricular block; four (4/10) patients had an intraventricular conduction disturbance. At the time of electrophysiologic study, two patients had a control spontaneous sinus cycle length that exceeded 1000 msec. Following propranolol, the mean spontaneous cycle length increased by 17.4% (924 to 1085 msec, P less than 0.005) and spontaneous second degree SA block reappeared in the one patient. The maximum escape cycle ranged from 116% to 229% of the prepacing spontaneous cycle length and was considered to be prolonged in two of ten patients. Propranolol had no significant effect on the maximum escape cycle/prepacing cycle length X 100 (%). The estimated sinoatrial conduction time (SACT) was determined in seven patients and ranged in value from 120 to 238 mes. Propranolol increased the mean value of the estimated SACT from 179 to 213 msec, P less than 0.025. Propranolol may cause marked bradyarrhythmias in some patients with sinus node dysfunction, and should be used with caution in these patients.


The conduction system in double outlet right ventricle with subpulmonic ventricular septal defect and related hearts (the Taussig-Bing group). The Taussig-Bind group of hearts is that group in which the aorta emerges completely from the right ventricle, unrelated to the position of the ventricular septal defect (VSD), while the pulmonary trunk emerges at varying degrees from the right ventricle related to the location of the VSD. Several types of surgical procedures involve the VSD in these hearts. The conduction systems in six of these hearts were therefore studied. The atrioventricular (A-V) bundle lies in the inferior (posterior) wall of the defect on the left side beneath the summit of the ventricular septum. In some cases, the bundle may be proximal to the defect, thus lying intramuscularly, and only the right and left bundle branches are related to the defect. The pattern od development of the atrioventricular (A-V) node, bundle and bundle branches is related primarily to the development of the endocardial cushions and posterior ventricular septum, and not to that of the absorption of the bulbus.


Nitroprusside after open-heart surgery.

Nitroprusside after open-heart surgery. The effects of intravenous infusion of sodium nitroprusside were studied in 11 children immediately after open-heart surgery for congenital heart disease. The patients were selected because, following bypass, their cardiac index was below 2,0 L/min/m2 and their systemic vascular resistance exceeded 30 units. In order to eliminate the effects of preload, mean left atrial pressure was maintained at a constant level by blood transfusion. During infusion of nitroprusside the mean decrease of mean arterial pressure was 18.6%, of systemic vascular resistance was 53.7%, and the increase in cardiac index was 76.9%. All children recovered.


Hemodynamic effects of vasodilator agents in dogs with experimental ventricular septal defects. The ratio of pulmonary to systemic vascular resistance (Rp/Rs) largely determines the amount of left-to-right shunting and pulmonary to systemic flow rat (Qp/Qs) in the presence of a large isolated ventricular septal defect. The possibility that pharmacologic reduction of systemic vascular resistance with alpha-adrenergic receptor blockade or beta-adrenergic receptor stimulation would increase the ratio Rp/Rs, and therefore reduce the ratio Qp/Qs, was studied in dogs in which ventricular septal defects had been surgically created. Administration of phentolamine and phenoxybenzamine caused a 42% reduction in Rs and no reduction in Rp. Qs was unchanged and Qp declined by 24% and the ratio Qp/Qs fell by 32%. Infusion of the beta-adrenergic receptor stimulant isoproterenol also reduced Qp/Qs. However, this was accomplished as a result of an increase in Qs and at the expense of an increase in heart rate. As a decline in the ratio Qp/Qs has been shown to be beneficial to patients with large left-to-right shunts, pharmacologic reduction of systemic vascular resistance may prove to be helpful in treating congestive heart failure in those patients with large left-to-right shunts at the ventricular level who are refractory to the usual decongestive measures.


Impaired forearm oxygen consumption during static exercise in patients with congestive heart failure. In this study, the effects of forearm static exercise were determined on local blood flow and oxygen consumption in 15 normal individuals (NL) and their responses were compared with ten patients in congestive heart failure (CHF). Forearm blood flow was determined by a plethysmographic technique before and during 15% of maximum voluntary contraction of the forearm. Regional arterial and venous oxygen contents were sampled and forearm oxygen consumption calculated by the Fick principle. At rest, forearm blood flow was less in patients with heart failure than in normal individuals;  however, this was compensated for by an increased oxygen extraction, thus maintaining forearm oxygen consumption at a normal level. In contrast, during static exercise, forearm blood flow failed to rise normally with heart failure (NL 9.31; CHF 4.35 ml/min-100 ml, P less than 0.001) and the increased oxygen extraction was not sufficient to maintain a normal forearm oxygen consumption (NL .82; CHF .44 ml/min-100 ml, P less than 0.01). Therefore, patients with congestive heart failure demonstrate regional circulatory and metabolic abnormalities during static exercise that are comparable to those present during dynamic exercise. Because of a limited ability of their skeletal muscle resistance vessels to respond to dilator stimuli, they have an attenuation of their exercise hyperemia which leads to an earlier shift to anaerobic metabolism.


Mitral valve area in combined mitral stenosis and regurgitation. Eight patients with mixed mitral stenosis and regurgitation underwent hemodynamic and angiographic study prior to mitral valve replacement. The stenotic orifice of the mitral valve was calculated employing the total left ventricular stroke volume by cineangiography as the numerator of the Gorlin Formula. Excellent agreement with the measured orifice of the mitral valve was obtained using a value of 37.9 (0.85 X 44.5) for the constant in the Gorlin formula as recommended by Cohen and Gorlin. Recalculation of this constant independently by our data yielded a value that was almost identical. Regurgitant flows and orifice sizes were calculated for each patient using the same constant as for calculation of the stenotic orifices.


Retrograde coronary artery flow in aortic valve disease. Retrograde coronary artery flow was observed angiographically in 43 patients with aortic stenosis and/or regurgitation. In the 24 patients with pure or predominant aortic stenosis, retrograde flow was seen in all 24 during end-systole. In the eight patients with pure aortic regurgitation, retrograde flow was seen mainly during end-diastole (6/8). Among the 11 patients with stenosis and regurgitation, retrograde flow was both end-systolic and enddiastolic. Dominant left coronary arteries were seen in 13 patients; 13 showed retrograde flow in the dominant arteries. Dominant right coronary arteries were seen in 25 patients: all 25 showed retrograde flow equally in the right and left coronary. Five of the 43 patients could not be evaluated for dominance because of coronary artery occlusions. The severity of retrograde flow did not correlate with usual clinical, hemodynamic or tension-stress parameters: angina, electrocardiographic abnormality, end-diastolic pressure or volume, end-systolic pressure or volume, ejection fraction, severity of aortic regurgitation, peak or mean valve gradient, aortic valve area, myocardial tension and stress calculations, or DPTI:SPTI. In summary, retrograde coronary artery flow was seen in all 43 patients with severe aortic valve disease. The time in the cardiac cycle when retrograde flow occurred was related to the type of valve disease. Retrograde flow was seen mainly in the coronary arteries supplying the left ventricle and may result from increased regional myocardial stresses.


Echocardiography of cardiac valves in pericardial effusion. Nine adult patients with large pericardial effusions (PE) demonstrated echocardiographic motion abnormalites of cardiac valves in systole. In four cases (Group 1), the abnormal findings consisted of prominent systolic anterior movements of the mitral valve resembling those seen in idiopathic hypertrophic subaortic stenosis. In Group 2(five cases), typical mitral valve prolapse patterns with large posterior midsystolic displacements well below the C point were observed. Additional abnormalities in Group 2 included tricuspid valve prolapse patterns (four cases), early systolic movement of the aortic valve toward closure (three cases), midsystolic notching of the pulmonary valve (two cases) and abnormal or attenuated motion of the aortic root in all patients. Marked decrease or resolution of PE resulted in complete disappearance of all the observed abnormalities. These findings appeared to be related to large fluid collections behind the left atrium and abnormal movement of the heart in the pericardial space. In the presence of PE, therefore, the echocardiographic observation of abnormal valve motion may not be clinically significant.


Echocardiographic assessment of the level of cardiac compensation in valvular heart disease. The level of cardiac compensation in valvular disease was studied by relating echocardiographic and cardiac catheterization measurements. Three groups -- compensated, intermediately compensated, and decompensated -- were defined according to the left ventricular angiographic pattern and cardiac output. The echocardiographic ejection indices, percent left ventricular minor diameter shortening, ejection fraction, and fiber shortening rate were significantly higher than normal in compensated mitral regurgitation, lower then normal in compensated aortic stenosis, and within normal limits in compensated aortic insufficiency. In the decomposed state these indices were depressed. Intermediate compensation was best recognized by combining several echocardiographic variables into an echocardiographic score based on multivariate discriminant function analysis. Thus, the compensated volume overload states (aortic and mitral regurgitation) and pressure overload state (aortic stenosis) have separate sets of "normal" echocardiographic values; low ejection indices characterize the decompensated group, while recognition of intermediate compensation requires analysis of multiple echocardiographic variables.


Coronary atherosclerosis in subjects with mitral stenosis. The coronary arteries were examined in 60 specimens from patients with mitral stenosis. In three, localized obstruction was nonatherosclerotic in nature (in one, arterial dysplasia; in two, embolic). In 18 of the remaining 57 cases (31.5%), significantly obstructive atherosclerosis in one or more segments of the coronary arterial system was found. This represented 46% of the males and 27% of the females. The incidence of involvement of three or more arteries by significantly obstructive atherosclerosis was 39%, while in a cited series of subjects with angina pectoris three or more vessels were involved in 79% of the cases. It may be concluded that, on the average, the distribution of lesions in patients with mitral stenosis and significant coronary atherosclerosis is less wide than in subjects with clinical coronary disease.


Studies on porphyrin biosynthesis in lead-intoxicated rabbits. The present report deals with studies on porphyrins and porphyrinogen carboxy-lyase of red cells and urinary porphyrins from lead-intoxicated rabbits. It was shown that the free erythrocyte porphyrins are mixture of protoporphyrin 9, the main component, and minor proportions of Coproporphyrin, Uroporphyrin III and Phyriaporphyrin. Analysis of the urinary porphyrins deomonstrates the presence of Coproporphyrins III as the major component, together with 15-20% of other porphyrins: 10-14% 5-COOH, 1-2% 6-COOH, 2-3% 7-COOH porphyrin and 1-2% Uroporphyrin III. We have not been able to detect an increase of Uroporphyrin I. Assays of porphyrinogen carboxy-lyase activity in hemolysate supernatant using Uroporphyrinogen III and Phyriaporphyrinogen (Phyria'gen) III as substrates, showed the existence of a slight decrease of both decarboxylase activities, being more affected during the second stage, the Phyria'gen decarboxylation. A possible regulation mechanism responsible for the porphyrin picture is discussed.


Quantitative determination of granulocytic amino acids in healthy men and women. The concentration of the amino acids has been determined in granulocytes from healthy men and women by chromatographic analysis on ion-exchanging resins. Except taurine, ornithine and aspartic acid, the concentrations of all the amino acids are greater in the cells isolated from women. The most variable amino acids are alanine, valine, metionine, isoleucine, leucine, tyrosine, phenylalanine, lysine and histidine. Taurine represents 50% of the pool determined from men, whereas for women it represents only 25%.


Solid-phase, magnetic particle radioimmunoassay. A solid-phase radioimmunoassay system has been developed based on the use of antibodies covalently linked to polymer-coated iron oxide (EnzacrylR). An electro-magnet is employed both to mix the particles during incubation (by switching the field on and off) and to separate the antibody-bound and free fractions. This obviates the need for vertical rotation and for the time-consuming, multiple centrifugations required with conventional solid phase procedures. The system is universally applicable and methods have been established for the assay of thyroxine, human placental lactogen and digoxin. The thyroxine assay was employed as a model and it was shown that the results obtained for serum samples correlated closely with those using a routine liquid-phase radioimmunoassay. The applicability of employing a second antibody linked to the iron oxide particles was also studied.


The relationship between erythrocyte superoxide dismutase activity and erythrocyte copper levels in normal subjects and in patients with rheumatoid arthritis. A quantitative assay for erythrocyte superoxide dismutase activity using the xanthine-xanthine oxidase-nitro blue tetrazolium system which is applicable to clinical material is described. The in-batch precision of the method is 3.5% and the between-batch precision is 8.8%. Employing this assay, erthrocyte superoxide dismutase activities were measured in 50 normal subjects and in 50 patients with rheumatoid arthritis. There was no significant correlatiion between superoxide dismutase activity and erthrocyte copper concentration. Erythrocyte copper was lower in female rheumatoid patients than in normals. This difference was not accompanied by a difference in superoxide dismutase activity.


A rapid and sensitive spectrophotometric procedure for the micro determination of carbamazepine in blood. An ultraviolet spectrophotometric procedure for the micro determination of carbamazepine in blood is described which is based on the original 9-methyl-acridine method proposed by Beyer, K.H. and Klinge, K. (1969) (Arzneim-Forsch. 19, 1759--1760). Carbamazepine is extracted from blood with dichloromethane, which is then washed with alkali and acid. An aliquot of the extractant is evaporated to dryness and the residue heated briefly with hydrochloric acid at 150 degrees C. Following removal of non-specific interference with n-heptane, the absorbance of the acid catalysed rearrangement product (9-methylacridine) is determined at 258 nm. The resulting procedure is rapid, reliable, sensitive and specific. It requires 100-200 mul sample for a single estimation and has a detection threshold of less than 0.1 mg/100 ml. It is concluded that the method is suitable for routine clinical use.


A rapid spectrophotometric procedure for the simultaneous micro determination of carbamazepine and 5, 5-diphenyl-hydantoin in blood. Carbamazepine and 5,5-diphenylhydantoin are simultaneously extracted from 100-200 mul blood with 1,2-dichloroethane. 5,5-Diphenylhydantoin is removed by a one-step wash into alkali. The dichloroethane is further washed with acid and then evaporated to dryness. 5,5-Diphenylhydantoin is determined in the alkali washing by a benzophenone procedure; carbamazepine is determined in the dried residue by the 9-methylacridine procedure earlier described. The combined method is rapid, reliable and has a detection threshold of less than 0.1 mg/100 ml for each drug.


Resting muscle levels and the influence of exercise on serum amino acid-arylpeptidase activity in man. Activities of amino acid arylamidases in skeletal muscle and serum of healthy male subjects were investigated using various substrates (Ala-, Leu-, Phe- and S-benxylcysteine-4-nitroanilides). At rest, there is no agreement between muscle and serum activity patterns. After physical exercise, the serum increase of Ala-, Leu- and Phe-AP activities is closely correleated to their respective activities in resting muscles. Since there is no relationship of the postexercise increase of amino acid arylpeptidase activitiy and exercise-induced intrasvascular haemolysis, it appears highly probable that the origin of the serum activity increase after physical work is skeletal muscle.


Cerebrospinal fluid and plasma glutamine elevation by anticonvulsant drugs: a potential diagnostic and therapeutic trap. Administration of phenobarbitone or primidone can produce an elevation of glutamine and ornithine concentrations, together with a reduction of urea concentrations, in the fasting plasma and CSF of some infants, and in the CSF of some older epileptic patients. These two anticonvulsant drugs may interfere with the metabolic conversion of ammonia to urea, possibly by enzyme inhibition at the step of carbomoyl phosphate synthesis. The elevation of glutamine in plasma and CSF can mimic values found in some hyperammonemias. Recognition of this drug reaction can prevent erroneous diagnoses of genetically-determined urea cycle disorders and subsequent inappropriate therapy.


[A modification of Laurell's one-dimension electrophoresis using polyvalent antisera (author's transl)]. A modification of the Laurell method is described which makes it possible to measure very small amounts of proteins. The use of the intermediate gel technique allows the simultaneous determination of two or more proteins by one-dimension electrophoresis. The method is useful for differentiation of organ-specific proteins from serum proteins, for identification of identical and not-identical antibodies in antisera and for checking protein products and antisera for their purity.


Quantitative determination of unconjugated pterins in urine by gas chromatography/mass fragmentography. A gas chromatographic/mass fragmentographic method is described which permits the determination of unconjugated pterins in urine. After the addition of 6,7-dimethylpterin as an internal standard, the acidified urine samples are purified by liquid chromatography on Dowex-50 and Dowex-1 columns. The pterins are then converted to their corresponding trimethylsilyl derivatives and the base peaks of biopterin (m/e 409), neopterin (m/e 409) and 6,7-dimethylpterin (m/e 320) are determined. The method is sensitive and specific and permits the processing of large numbers of samples. By means of this method, the urinary excretion of biopterin and neopterin from 9 healthy subjects has been determined.


Lipid composition of bile in morbid obesity before and after jejunoileal bypass surgery. Bile cholesterol, phospholipids, total bile acids, individual bile acids, and fatty acid compositions of bile neutral lipids and phospholipids were analyzed before, at one month and at six months following jejunoileal bypass surgery in a series of morbidly obese patients. Preoperative mole percentages of cholesterol and lithogenic indices were high, indicating that biles were supersaturated with cholesterol and outside the micellar solubility zone when plotted on triangular coordinates. At the one month post-operative period percentages of cholesterol and lithogenic indices were significantly increased as compared to the pre-operative state. At six months post-operatively these values had decreased to approximately the pre-operative levels. No changes were observed in percentages of lithocholic acid, but deoxycholic acid decreased to markedly low levels at one month and remained low at the six month post-operative interval. Relative proportions of cholic acid increased, and the ratio of cholic to chenodeoxycholic acid was significantly increased at both post-operative intervals. No significant changes were noted in bile neutral lipid or phospholipid fatty acid composition, indicating that no depletion of essential fatty acids had occurred.


A modified ultraviolet spectrophotometric method for the determination of theophylline in serum in the presence of barbiturates. Ultraviolet spectrophotometry is the most commonly used technique for the determination of theophylline levels for therapeutic monitoring. Common interferences in most methods are barbiturates and xanthines. A modified method which eliminates interferences from barbiturates and most xanthines is presented. Theophylline is extracted from serum with chloroform/isopropanol at pH 7.4 back extracted into dilute hydrochloric acid and then the solution is made alkaline with sodium hydroxide. Barbiturate interferences are eliminated. Interference from caffeine, uric acid, 7-(2,3-dihydroxypropyl)theophylline, xanthine, and hypoxanthine are not observed. Theobromine, and the metabolite 3-methylxanthine interfere. Interference is not observed from quinidine, diazepam, salicylate, glutethimide, methylprylon, propranolol, methaqualone, dilatin and ethchlorvynol. Sulfanilamide, procainamide and chlordiazepoxide interfere.


Radioimmunoassay for 3,3',5'-triiodothyronine (reverse T3, R-T3) in unextracted human serum. Highly specific antibodies against 3,3',5'-triiodothyronine (reverse T3, R-T3) have been produced in rabbits. The crossreaction with T4 is about 0.05%. A radioimmunoassay for R-T3 in unextracted serum was developed. ANS is used for blocking the binding of tracer and endogenous R-T3 to TBG. The sensitivity to the assay is 0.06 ng/ml plasma. The mean normal R-T3 concentration is 0.20 ng/ml. Thyrotoxic patients show elevated levels; in most hypothyroid patients R-T3 concentrations are below the detection limit.


The use of electrolyte measurements in the detection of cystic fibrosis. Six tests recommended for use in the diagnosis of cystic fibrosis (CF) have been compared in the same subjects. The tests were carried out on 165 normal persons, 64 known cases of CF, their 67 parents and 18 sibs. The tests measured sodium in fingernails by activation, sodium in saliva and chloride in saliva and in thermal and pilocarpine-induced sweat by means of ion-specific electrodes, and chloride in pilocarpine-sweat by the standard titrimetric method. The tests ranged in complexity from simple screening methods to individual clinical procedures. None of the simpler tests matched the standard pilocarpine method in diagnostic efficiency, whether they were used singly or in combination.


The simultaneous determination of lidocaine and procainamide in serum by use of high pressure liquid chromatography. This report describes an accurate, sensitive rapid procedure for the determination of lidocaine and procainamide at therapeutic concentrations in serum. The drugs and an added internal standard (procaine) are extracted from serum using charcoal adsorption. The analysis is carried out by high pressure liquid chromatography on a reverse-phase column using buffered aqueous acetonitrile as the mobile phase. Chromatography is complete in 10 min. C.V. values of 10% and 6% for concentrations of 1 mg/l and 20 mg/l, respectively, are attainable routinely. Total analysis time is 15 min.


Ganglioside pattern and neuraminic acid content of human gastric and colonic carcinoma. Twenty-five human gastric and 11 human colonic adenocarcinomas were analysed for their ganglioside pattern and for their content of lipid-bound and protein-bound neuraminic acid. In most carcinomas the content of both lipid-bound and protein-bound neuraminic acid was increased by an average of four- and two-fold, respectively. The ganglioside pattern of all the carcinomas resembled that of normal tissue. In six gastric carcinomas the content of lipid-bound neuraminic acid and the ratio of lipid-bound neuraminic acid to protein-bound neuraminic acid (L/P ratio) were lower than those of normal gastric mucosa. These carcinomas were significantly larger than the rest of the tumours.


The spectrophotometric determination of uroporphyrinogen I synthetase activity. A simple spectrophotometric method for uroporphyrinogen I synthetase in erythrocytes is described. Results obtained on intermittent acute porphyria patients and carriers are similar to the results obtained with fluorimetric methods. Reproducibility, relationship between enzyme activity and enzyme concentration, and effect of time on enzymatic activity are described.


Limits of the estimate of serum folate-binding protein by radioassay. Various factors that influence the measurement of serum folic acid-binding protein (FABP) by radioassay were examined. Varying the protein concentration and the composition of the charcoal suspension can increase or reduce the value of the blank, i.e. the estimate of FABP. Moreover, it was found that the value of the FABP depends on the quantity of folic acid in the incubation medium. Reproducible results can only be obtained with rigorously defined conditions of concentration of the different reactants.


Zinc and copper in human semen.

Zinc and copper in human semen. Investigations have been performed on the possible effect of zinc and copper on the fertility of man. Semen was obtained from 67 men and the zinc concentration was found to be decreasing with a drop of the activity of the semen. Copper was found in semen in microgram amounts which do not change with the activity of the semen. Results suggest that the zinc concentration may serve as evidence in the evaluation of semen activity.


Comparison between human 125I-labelled TSH labelled with lactoperoxidase or chloramine-T; advantages in the use of enzymically labelled antigen in a radioimmunoassay for human TSH. 1. Human thyroid stimulating hormone (human TSH) was iodinated by both chloramine-T and lactoperoxidase methods. 2. Human 125I-labelled TSH prepared by the enzymic method showed less damage on storage, an increased shelf-life, and improved binding at different antiserum dilutions. 3. A comparison of the values of the bound counts on the addition of human TSH, expressed as a percentage of the zero standard bound counts (B/BO values) in the standard curves, demonstrated that lactoperoxidase-labelled human TSH gave increased sensitivity.


Quality control in haemoglobinometry with special reference to the stability of haemiglobincyanide reference solutions. In haemoglobinometry grave errors are still being made even though an internationally accepted standardized method is available for the determination of the haemoglobin content of blood. Up to the present only haemiglobincyanide reference solutions have been available on a wide scale to check the measuring stage of the standardized haemiglobincyanide method. These reference solutions are shown to remain stable, under proper storage conditions, for more than 10 years. Concentrated haemoglobin solutions have become available recently, offering the possibility to control the dilution and conversion steps of the haemiglobincyanide method. Such a solution is shown to remain stable, under proper storage conditions, for at least a year. Using both haemiglobincyanide reference solutions and concentrated haemoglobin solutions, as well as having the possibility of checking the cyanide content of the reagent used, an acceptable intra-laboratory control program may now be set up.


The occurrence and identification of o-hydroxyhippuric acid (salicyluric acid) in the urine of sick children. O-Hydroxyhippuric acid has been identified in the urine of a number of sick children who were not receiving salicylate-containing drugs. Characterisation was effected by thin-layer and gas-liquid chromatography and by gas-liquid chromatography-mass spectrometry. An association between gastro-intestinal dysfunction and the excretion of o-hydroxyhippuric acid was observed in approximately 50% of the patients who excreted this substance.


Biological and analytical variation of commonly determined blood constituents in healthy blood donors. The relative contributions of analytical error and non-systematic biological variation to the reference range for 19 commonly-determined blood chemistry values have been estimated during the course of studying 1000 healthy blood donors. Estimates of non-systematic biological variations are arrived at by determining the variation found in donors matched for factors known to affect blood chemistry such as age and sex, and deducting the within-batch analytical error which applied. Comparison of the within-batch and between-batch analytical precision with the biological variation showed that the percentage increases in the normal range attributable to between-batch analytical error varies between 1 and 29%. This contribution is particularly large in the case of the electrolytes and amounts to no less than 29% in the case of calcium. Apart from well-known relationships a remarkable lack of correlation between the concentration of the different constituents in health is demonstrated.


A method for the estimation of 2-phenylethylamine in human urine by gas chromatography. A gas chromatographic procedure with flame ionisation detection has been developed to measure 2-phenylethylamine in human urine as the trifluoroacetyl derivative. The amine is purified by an ether extraction procedure and derivatised with trifluoroacetic anhydride. Quantification is obtained by comparing the peak height with that for an internal standard added to the urine before extraction. The normal 24-h excretion value for 2-phenylethylamine in human urine is found to be 6.8 +/- 2.9 mug (mean +/- S.D.).


A simple and rapid thyroxine radioimmunoassy (T4-RIA) in unextracted human serum; a comparison of T4-RIA and T4 displacement assay, T4(D), in normal and pathologic sera. A simple, rapid and accurate thyroxine radioimmunoassay (T4-RIA) in unextracted serum or plasma has been described, and for comparison T4 determinations have also been made by a T4(D) procedure using Abbott Tetrasorb kits. T4-RIA procedure basically involved denaturation of serum to dissociate T4-protein bond, and T4 released was allowed to react with [125I]T4-labeled T4 antiserum elicited by immunizing rabbits against bovine thyroglobulin. The displaced unbound [125I]T4 was rapidly taken up by an anionic resin sponge within 15 min and this sponge [125I]T4 uptake was linearly related to T4 present in standards or serum. The denaturation of serum effected by trichloroacetic acid-sodium hydroxide permitted virtually 100% T4 extraction recovery in normal, pregnancy, hypo- and hyperthyroid sera whereas 72.9-87.6% T4 recovery from normal serum (and with large individual differences) was noted with lower alcohols in T4(D) procedure. Cumbersome and/or tedious steps such as pre-extraction, centrifugation, time consuming bound and unbound hormone separation procedures, etc. are obviated in T4-RIA and the entire assay can be completed in the same tube in approximately an hour. These attributes along with increased sensitivity and specificity and the need for only microamounts of test sera (25-50 mul) in T4-RIA offer distinct advantages over T4(D) procedures, and in simplicity excel even other T4-RIAs. T4-RIA values in physiological and pathological states were highly correlated (r = 0.97) with T4(D) measurements and no differences between these two techniques were found. The reported discrepancies between T4-RIA and T4(D) measurements in human sera and some of the reasons for attributing these inconsistencies to probable methodological errors and variations are discussed.


A simple and sensitive assay of total serum bile acids. A simple and sensitive method was developed for the quantification of serum total 3alpha-hydroxy bile acids. 0.1 ml of serum was mixed with tris(hydroxymethyl) aminomethane hydrochloric acid buffer and heated at 67 degrees C for 30 min. To the solution were added 3alpha-hydroxysteroid : oxidoreductase (EC 1.1.1.50; 3alpha-HSD), NAD, diaphorase (EC 1.6.4.3) and resazurin. The mixture was incubated at 20 degrees C for 1 h. The resultant fluorescence of resorfin was measured at 580 nm with the excitation at 560 nm. The blank value was obtained after the same treatment of another 0.1 ml of the same serum without 3alpha-HSD. A linear relationship was obtained between the amount of bile acids and the fluorescence intensities in the range of 1 to 150 mumol/1. The recovery of bile acids added to the serum was 81.4 +/- 2.5 (S.D.)% for cholate, chenodeoxycholate and deoxycholate. The bile acid content in the serum was 48.8 mumol/1 with a standard deviation of +/- 0.42 and a coefficient of variation of +/- 0.87% in 10 replicate determinations. The mean bile acid content of normal fasting male sera was 8.0 mumol/1 (3.6-12.6 mumol/1, n = 12) and of female sera 6.8 mumol/1 (3.2-12.7 mumol/1, n = 13).


A simple radioassay for 25-hydroxycholecalciferol without chromatography. A simple method to assay 25-hydroxycholecalciferol in human plasma without chromatography is described. The method includes ammonium sulphate precipitation of 25-hydroxycholecalciferol bound to plasma-transport globulins as a purification step, extraction with toluene and saturation analysis utilizing human osteomalacic plasma for competitive protein-binding assay. The criteria of reliability of the method are reported.


A routine assay procedure for monoamine oxidase and its application to human blood platelets. In the radiometric assay procedure for monoamine oxidase separation of the reaction products by ion-exchange column chromatography was optimized. A device was constructed that allowed the separation of 48 samples at the same time. This device can be applied for a great variety of analytical methods. With a 0.1 mM substrate concentration in the incubation mixture, reaction products could be determined with a reproducibility of 2-5%, where the standard deviation depends on the substrate. Using beta-phenylethylamine as substrate 20 mug of platelet protein was sufficient for monoamine oxidase activity determinations. The dependence of human blood platelet monoamine oxidase on age and sex were studied using the method.


A method for galactose-1-phosphate uridyltransferase assay and the separation of its isozymes by DEAE-cellulose column chromatography. A simplified radioactive assay for galactose-1-phosphate uridyltransferase (EC 2.7.7.12) using a small DEAE-cellulose column for the identification of the endproduct (uridine diphosphate galactose) is described. The enzyme activities in red blood cell hemolysates of normal subjects are in the range of 24 to 33 (average 30.3) mumol UDPgalactose produced per g hemoglobin per h and in fibroblasts 0.39 and 1.39 (average 0.71) nmol per mg protein per min. Furthermore, different isozymes of red blood cell galactose-1-phosphate uridyltransferase were separated on DEAE-cellulose columns. In the case of a normal genotype, most of the enzyme activity is eluted at the earlier fractions with the low molar phosphate buffer, whereas the Duarte variant appeared at later fractions with higher molar phosphate buffer.


Electrophoretic modifications of three enzymes in extracts of human and bovine lens. Posttranslational "aging" of lens enzymes. Electrophoretic modifications have been found in extracts from human and bovine lenses for three enzymes: glucose-6-phosphate dehydrogenase, triosephosphate isomerase and nucleoside phosphorylase. Increased anodic mobility is observed in all cases. It is more pronounced than in red cell lysates, also more evident in lenses from adult than from young animals. These results give evidence of posttranslational "aging" of enzyme molecules in lenses.


Determination of zinc in fingernails by non-flame atomic absorption spectroscopy. The determination of Zn in fingernails directly using the graphite furnace presented certain difficulties due to the anomalous behavior of the analyte in the furnace. The appearance of two peaks which were due to Zn and not to any background interference was noted. The Zn value obtained by adding the area of these two peaks compared fairly well with Zn levels determined by wet ashing and subsequent determination either in the furnace or flame. Wet ashed samples gave only a single peak. It was possible to produce a model of the phenomenon with various Zn salts in a non aqueous matrix. Under these conditions ZnSO4 and ZnO gave a discretely different peak from ZnC12 or metallic Zn. Several tissues such as serum, whole blood, cuticle, and hair were examined for multiple peak formation. Direct determination of Zn in fingernails with the graphite furnace is possible for certain applications such as the determination of Zn levels of white spots in fingernails. For this purpose it is possible to use a sample size as small as 20 mug using the 2138 Zn line. This allows one to run several determinations on a single white spot. However, where sample size is not a limitation, wet ash digestion prior to determination in the furnace is probably the preferred procedure.


3-Hydroxy-3-methylglutaric aciduria, combined with 3-methylglutaconic aciduria. 1. A baby with severe metabolic acidosis was found to excrete abnormal amounts of 3-methylcrotonic acid, 3-methylglutaconic acid and 3-hydroxy-3-methylglutaric acid in urine. 2. Several other abnormal constitutents appear to be products of side-reactions and include 3-hydroxy-3-methylbutyric acid and 3-methylglutaric acid. 3. The profile of acids in urine indicates a blockage in the sixth step of leucine catabolism, the cleavage of 3-hydroxy-3-methylglutaryl-CoA to acetoacetic acid and acetyl-CoA.


Identification of beta-aminoisobutyric acid in uremic serum. An unidentified ninhydrin-positive substance found in uremic sera but not found in normal sera was isolated by gel-filtration through Sephadex G-75 followed by high voltage paper electrophoresis (pH 3.5), and identified as beta-aminoisobutyric acid using paper chromatography and automated amino acid analyzer. The quantitative determination of beta-aminoisobutyric acid in serum revealed that the level of beta-aminoisobutyric acid in uremic sera was much higher than that of normal sera. Gas chromatographic determination of the enantiomorphs of beta-aminoisobutyric acid showed that uremic sera contain R- and S-isomers of the amino acid, but with the R-isomer as the dominating form.


Suberylglycine excretion in the urine from a patient with dicarboxylic aciduria. Suberylglycine (HOOC(CH2)6CONHCH2COOH) was found in the urine from a patient with C6-C10-omega-dicarboxylic aciduria and unexplained episodes of lethargy and unconsciousness. The total excretion of adipic, suberic and sebacic acid ranged from 0.77 to 1.3 mg/mg creatinine after episodes of acute attack of the disease. Suberylglycine, identified by gas chromatography/mass spectrometry, was repeatedly found in the urine samples. The amount of this conjugate ranged from 0.2 to 0.5 mg/mg creatinine. The precursors of the dicarboxylic acids are suggested to be long chain monocarboxylic acids, oxidized through omega- and beta-oxidation to adipic, suberic and sebacic acid. Suberylglycine is subsequently formed by glycine-N-acylase catalyzed conjugation.


Factors involved in the determination of triglycerides in serum: an international study. Ten laboratories analysed five different specimens in duplicate on ten separate occasions by one, or several, of five common triglyceride methods. Simple statistical data are presented and, as far as possible, these are interpreted in the light of the methods used and the results of chemical analyses of the materials. Great variability was found between the results of the participating laboratories. The major factor involved seems to be the material specific nature of the methods under study. At least one method in common use is contraindicated.


Phosphatidylcholine content and fatty acid composition of tracheal and gastric liquids from premature and full-term newborn infants. The phosphatidylcholine content and fatty acid composition of tracheal and gastric liquids from newborn full-term and premature (30-36 weeks) infants were studied. Phosphatidylcholine accounted for about 70% of the total phospholipids in both liquids from the full-term infants but only 55-57% of those from the prematures. There was a significant correlation between tracheal and gastric liquids in the fatty acid composition of phosphatidylcholine. There were significant differences in fatty acid composition between the full-term and premature liquids. These differenences were most apparent in the gastric liquid. It is suggested that phospholipid or fatty acid analysis of tracheal or gastric liquids from newborn infants can be used in the assessment of pulmonary maturity.


Plasma dopamine beta-hydroxylase in a noradrenalin-secreting pheochromocytoma. Circulating dopamine beta-hydroxylase activity was studied in a case of a hypertensive patient bearing a pheochromocytoma. The tumour secreted noradrenalin. The enzyme activity was found to decrease gradually after removal of the tumour, and to reach a stable value, in correlation with the decrease of urinary catecholamines. It is concluded that some pheochromocytoma tumours are able to secrete dopamine beta-hydroxylase.


Isoelectric focusing of serum in cystic fibrosis: failure to distinguish between homozygote and heterozygote sera. Thin-layer isoelectric focusing was performed on samples of sera from patients with Cystic Fibrosis, siblings and obligate heterozygotes (parents), and children without cystic fibrosis (controls). The protein band with an isoelectric point of pH 5.48, previously reported to be absent in homozygote cystic fibrosis sera, was found to have a pI of 5.25. It was present in approximately one-half of the homozygote and heterozygote sera tested, and absent from 18 percent of control sera. The presence of this band is not therefore a reliable marker for the normal gene, and cannot be used to identify the heterozygous carrier for cystic fibrosis.


Selective inhibition of lymphocyte responsiveness to phytohaemagglutinin in patients with Reiter's syndrome. The influence of synovial fluid from four patients with Reiter's syndrome on lymphocyte responsiveness was studied. On synovial fluid was found which specifically depressed the responsiveness of lymphocytes from patients with Reiter's syndrome to the non-specific mitogen phytohaemagglutinin (PHA). The ratio of the responsiveness of lymphocytes cultured in the presence of foetal calf serum (FCS), compared to those incubated in the Reiter's synovial fluid, was used as a measure of the depression induced by the Reiter's synovial fluid. The mean ratio for eight normals stimulated with PHA was 0-70 (range 0-35-0-96), while for eight patients with Reiter's syndrome, it was 0-13 (range 0-07-0-19). Similar studies done with concanavalin A (con A) showed no difference between lymphocytes from normals (0-73) or patients with Reiter's syndrome (0-67). Chromatography of the Reiter's synovial fluid on a Sepharose 4-B column resulted in the separation of three major fractions, one of which exhibited the inhibitory activity. When this active fraction was absorbed with Reiter's lymphocytes, a loss of the inhibitory activity of the fraction was seen. A similar absorption with normal lymphocytes had no effect. These studies demonstrate that a factor present in the synovial fluid of a patient with Reiter's syndrome reacted specifically with lymphocytes from patients with Reiter's disease and not with lymphocytes from normals. The interaction of this factor with lymphocytes from patients with Reiter's syndrome inhibited the responsiveness of these lymphocytes to PHA but not to con A.


Reconstitution of defective cellular immunity with foetal thymus and dialysable transfer factor. Long-term studies in a patient with chronic mucocutaneous candidiasis. Extensive studies of a 9-year-old boy with recurrent pulmonary infections and chronic mucocutaneous candidiasis disclosed a severe defect in cell-mediated immunity but normal humoral immune responses. These immunological defects were not improved by initial treatment with transfer factor. After receiving a foetal thymus transplant the patient developed positive delayed-type skin tests, could be sensitized with chlorodinitrobenzene, and showed progressive improvement of in vitro lymphocyte functions including spontaneous formation of rosettes with sheep erythrocytes and positive responses to phytohaemagglutinin, concanavalin A and allogeneic leucocytes. Moreover, lymph node cellularity increased, especially in the thymus-dependent zones. Though the in vitro responses persisted for over 1 year, skin tests became unreactive at 38 weeks. However, in contrast to the pre-transplant experience transfer factor was now effective in inducing positive skin tests. These studies provide a chronological account of the effect of the thymus on expression of lymphocyte-mediated immune responses in man and suggest that thymus-derived cells are required for acquisition of transfer factor-induced cellular immunity.


Transient appearance of oligoclonal immunoglobulins and measles virus antibodies in the cerebrospinal fluid in a case of acute measles encephalitis. Acute measles encephalitis with severe sequelae in a 25-year-old man was studied. A transient appearance of oligoclonal IgG in cerebrospinal fluid and of intrathecally produced measles antibodies was found during 2 months after the onset of the disease. On the basis of this finding of local hyperimmunization it is proposed that in the case studied the measles virus infection may have been directly responsible for the disease process in the central nervous system.


Detection of receptors for immunoglobulin on human placenta by EA rosette formation. Direct evidence for the existence of Fc receptors on the surface of first trimester and term human placental cells has been obtained by the use of an antibody-coated red cell (EA) rosette assay. The modification of a Ficoll density gradient separation procedure for placental cell population in conjunction with dye uptake experiments, cytocentrifuge preparations and cytological analysis has enabled an identification of the rosette-forming cells in the mature placenta as predominantly, if not entirely, syncytiotrophoblastic. The significance of these findings, together with those demonstrating the presence of cell-surface Fc receptors on the chorionic membrane, are considered in relation to the transmission of immunoglobulin from mother to foetus and to the protection of the foetus as an intra-uterine allograft.


Cell-mediated anti-embryo cytotoxicity in human pregnancy. The microcytotoxicity assay has been used to study human maternal cell-mediated immunity against foetal lung cells derived from foetal autopsies and amnion cells from full-term deliveries. Cytotoxicity against semi-allogeneic foetal lung cells but not against adult skin fibroblasts was detected in fourteen cases. No cytotoxicity was detected in six mothers tested during the first 15 weeks of gestation whereas 58% of mothers in the 15-17 weeks of gestation (twelve cases) and 88% with more advanced pregnancies (eight cases) were specifically cytotoxic. Cross-reactions against allogeneic foetal lung cells were mainly produced by effector cells from patients with 16-17-week-old pregnancies. No cytotoxicity was detected against semi-allogeneic amnion cells by effector cells from mothers after full-term deliveries. Cytotoxicity was blocked by autologous post-abortum sera in two out of seven cases. The effect of autologous post-abortum sera on non-cytotoxic effector cells was nonspecifically arming in four out of ten cases.


Specific blastogenesis and lymphokine production in DNCB-sensitive human leucocyte cultures stimulated with soluble and particulate DNP-containing antigens. A soluble hapten (dinitrobenzene sulphonic acid (DNBSO3)) and particulate antigens consisting of DNCB, DNFB or DNBSO3 complexed with erythrocytes, all induce a specific blastogenic response and lymphokine production in leucocyte cultures from human subjects topically sensitized to dinitrochlorobenzene (DNCB). While low concentrations (50-100 mug/ml) of DNBSO3 could be left in human leucocyte cultures the entire 4 or 5 days of culture and result in reasonable levels of blastogenesis, it was found that consistently higher degrees of blast transformation resulted when DNCB-sensitive leucocytes were exposed to high concentrations of DNBSO3 (500 mug/ml) for a short period (2 hr). Cell-free supernatants from DNCB-sensitive leucocyte cultures harvested after 48 hr induced blastogenesis and DNA synthesis in secondary target leucocyte cultures from subjects not sensitized to DNCB. Such a blastogenic factor or lymphokine appeared to stimulate even in the absence of any residual antigen, since DNCB complexed to erythrocytes was removed by simple filtration through a 0-45 mum Millipore filter. In contrast to DNCB complexes, the antigenic activity of DNBSO3 complexed with erythrocytes was not removed by such filtration. Thus, several DNP-containing haptens (DNCB, DNFB, DNBSO3) induce specific lymphocyte transformation and lymphokine production when exposed in several different manners to leucocytes from humans sensitized to DNCB. The ability to use either a particulate or soluble stimulant in vitro offers a versatile system for studying cell-mediated immunity in humans with a broad range of potential applicability in both investigate and clinical medicine.


Primary immune response to Helix pomatia haemocyanin in malignant melanoma. Relationship between 19S and 7S antibody response and in vitro lymphocyte transformation. The primary immune response to Helix pamatia haemocyanin (HPH) was investigated in sixty-one patients with various clinical signs of malignant melanoma and compared to that of controls. Anti-HPH antibody response was only found abnormal in patients with widespread disease and visceral metastases, apparent from decreased 7S and increased 19S antibody levels. In vitro HPH-induced lymphocyte transformation, in contrast, was not only decreased in this late stage, but also in part of the patients in the beginning of the disease and was then correlated with subsequent tumour recurrence within 6 months. Generally, patients with positive lymphocyte transformation showed significantly higher 7S anti-HPH titres (mean 3-3 +/- 2-4 than patients without (1-7 +/- 1-6). In contrast to the lymphocyte transformation reaction, anti-HPH antibody response was not correlated with subsequent tumour recurrence. The same phenomenon of decreased 7S and increased 19S antibody response as in the melanoma patients with far advanced disease was observed to a less extent in controls with ageing. Anamnestic antibody responses to diphtheria and tetanus toxoid and serum IgG and IgA levels were found normal. Elevated serum IgM levels were more frequently observed in patients with localized (19/31) and disseminated disease (11/29) than in controls (2/31). These results suggest that melanoma patients develop impaired T-lymphocyte functions in final stages with visceral metastases. A previous study showed that minor defects in lymphocyte function in an early stage as measured by HPH-induced lymphocyte transformation have predictive value for subsequent tumour recurrence.


The effect of amino acid restricted diets on the clearance of 125I-labelled polyvinyl pyrrolidone in mice. Impaired overall phagocytic function, as measured by clearance of radio-labelled polyvinyl pyrrolidone from the blood (KPVP), has been demonstrated in mice whose dietary intake of the essential amino acids phenylalanine and tryptophan was reduced to less than a quarter of that in a standard diet which supported optimal growth. This was associated with loss in weight of body, liver, spleen and thymus and an increase in the phagocytic index, alphaPVP.


Plasma binding and the affinity of propranolol for a beta receptor in man. The effects of plasma drug binding on the relationship between propranolol concentration and the antagonism of isoproterenol tachycardia have been investigated in 8 normal subjects and 8 hypertensive patients. During chronic intravenous infusion of propranolol giving a narrow range of total plasma concentrations (22.5 to 50 ng/ml), there was, at best, a poor correlation with effects. On the other hand, there was excellent correlation between efficacy and free drug concentration, which fitted the predictions of the receptor theory of competitive antagonism. The true affinity constant for the binding of propranolol to its receptor can be calculated in terms of free drug concentration (KAfree) and was found to vary 2-fold compared to the affinity constant in terms of total plasma concentration (KAtotal) which varied 4-fold, the greater variation being due to plasma binding differences. Compared to normal subjects, KAfree and KAtotal were smaller in hypertensive subjects, implying lesser sensitivity to the drug, and plasma propranolol binding was greater. There was no difference in KAfree between high- and low-renin essential hypertensives, but KAtotal was smaller in the high-renin group due to increased plasma binding which did not reach statistical significance. It is concluded that the effect of propranolol on heart rate is a predictable function of free drug concentration in man and that the contribution of individual variation in receptor sensitivity to differences in oral dosage requirement is minor compared to that of variations in bioavailability.


Studies on digitalis. VII. Influence of nephrotic syndrome on protein binding, pharmacokinetics, and renal excretion of digitoxin and cardioactive metabolites. Serum protein binding of digitoxin was lower (p less than 0.05) in 7 patients with nephrotic syndrome (96.2%, SD 1.4) than in 51 control patients (97.3%, SD 0.5). Urine protein binding of digitoxin was 60.1% in the 6 nephrotic patients in whom it was determined. Simultaneous serum and urine measurements of digitoxin and cardioactive metabolites were performed in 5 patients after a single intravenous dose of 0.6 mg digitoxin. A modified 86Rb method was used. Mean T/2 of serum elimation was 4.8 days and 8.1 days in 5 control subjects (p less than 0.05). Serum concentrations 24 hr after the dose were lower in the nephrotic group (p less than 0.0025). The urine concentration T/2 with a mean value of 5.0 days was not significantly different from controls (7.2 days). The cumulative renal exeretion was higher in the nephrotic group (23.2% of dose) than in controls (15.8%) for 8 days. The excretion during one serum T/2 was the same in the two groups. Increased renal excretion thus explains the shortened serum T/2 in nephrotic patients. Preliminary data on the metabolic pattern of digitoxin and cardioactive metabolites in serum and urine suggested that drug metabolism may be changed in patients with nephrotic syndrome. As renal excretion is enhanced, patients with nephrotic syndrome will require higher doses of digitoxin. They should be maintained at lower than usual serum levels of total drug due apparent increased volume of distribution and hypoalbuminemia with consequent increased free drug fraction.


Effect of diazepam on ventilatory responses. To investigate the effects of diazepam on ventilatroy control, hypoxic and hypercapnic ventilatory responses were studied in 8 normal subjects before and after 10 mg of intramuscular diazepam. There was no significant change in either resting minute ventilation or resting end-tidal CO2 tension, but depression of hypoxic ventilatory response was observed 15 (60% of control) and 30 min (53% of control) after diazepam (p less than 0.05). No significant depression of hypercapnic ventilatory response was noted 70 to 130 min after diazepam. In view of the depression of hypoxic ventilatory response by diazepam in normal subjects, adverse responses along these lines should be considered in patients with impaired ventilatory function, such as chronic airways obstruction, and in those encountering acute hypoxemia.


Subjective and physiologic effects of morphine, pentobarbital, and meprobamate. These studies extend previous observations on the effects of pentobarbital on subjective states and postrotational nystagmus in postaddict subjects. Pentobarbital (150 mg) induced a degree of liking and an elevation of the morphine-benzedrine group (MBG) scale score equivalent to 24 mg of morphine. The effects of pentobarbital and meprobamate on postrotational nystagmus were studied using electro-oculography. Both drugs increased the frequency and prolonged the duration of postrotational nystagmus in a dose-related manner. Meprobamate was about 1/15 as potent as pentobarbital in enhancing postrotational nystagmus and producing signs of sedation.


Antipyrine: radioimmunoassay in plasma and saliva following administration of a high dose and a low dose. A simple and senstitive radioimmunoassay has been developed for the determination of antipyrine levels in plasma and saliva of man. Antiserum to antipyrine was obtained from rabbits immunized with an immunogen prepared by covalently coupling N-(4-antipyrinyl)-succinamic acid to bovine serum albumin (BSA). The radioimmunoassay can detect antipyrine levels as low as 10 ng/ml of plasma or saliva, using a 0.1-ml sample. This contrasts with the sensitivity of a commonly used spectrophotometric method that can measure about 4,000 ng/ml using a 2-ml plasma sample. Agreement between the radioimmunoassay and spectrophotometric assay of antipyrine was excellent for plasma (r = 0.98) and salvia (r =0.97) when samples were analyzed from 6 subjects receiving 18 mg/kg of antipyrine. The correlation between plasma and saliva antipyrine half-lives using the radioimmunoassay and an 18 mg/kg dose of antipyrine was r = 0.90 (p less than 0.005). After a dose of 1.8 mg/kg of antipyrine, the drug disappeared monoexponentially from plasma and saliva for at least 51 hr, and the correlation between plasma half-life and saliva half-life was r = 0.97 (p less than 0.001) in the 6 subjects. Excellent agreement was also observed between half-lives after the high and low doses of antipyrine (r = 0.99, p less than 0.001 for plasma and r = 0.98, p less than 0.001 for saliva).


Enzyme induction by oral testosterone.

Enzyme induction by oral testosterone. Six normal male volunteers ingested a dose of 400 mg free testosterone daily as tablets over 21 days. By the end of treatment intravenous antipyrine half-life had decreased significantly from 8.0 +/- 2.7 to 5.7 +/- 2.6 hr. The subjects eliminated testosterone from serum more rapidly on the twenty-first day of testosterone ingestion than on the first day. Serum albumin, bilirubin, prothrombin, alanine-amino-transferase, and alkaline phosphatases were unchanged during the experiment. It is concluded that oral testosterone treatment induces the hepatic drug-metabolizing system including that of testosterone.


Prevalence of ischaemic heart disease, arterial hypertension and intermittent claudication, and distribution of risk factors among middle-aged men in Moscow and Berlin. The initial examinations of prospective epidemiological studies of the same kind performed in random samples of 50-54 years old men from Moscow and Berlin showed the following prevalence rates (each first figure for Moscow, second for Berlin): ischaemic heart disease 14.4/13.6%, arterial hypertension 18.0/19.0%, intermittent claudication 6.9/3.4%, cigarette smoking 46.5/51.4%, overweight 22.9/21.9%, impaired glucose tolerance 36.6/42.5%. Hypercholesterolaemia (24.6/8.1%) was not comparable because different determination methods were used. In both population samples almost a half of IHD and 1/3-1/2 of hypertension cases were newly detected only by the study examinations. More than 80% of the 50-54 years old men showed the presence of one or more risk factors. The following significant relationships between disease groups and risk factors were found: for IHD, with hypertension, impaired glucose tolerance, and hypercholesterolaemia (Moscow only); for hypertension, with overweight, impaired glucose tolerance, and hypercholesterolaemia (Moscow only); for intermittent claudication, with cigarette smoking, impaired glucose tolerance, and hypercholesterolaemia (Moscow only). These comparative studies demonstrate the benefit and need of further common research and efforts for prevention and control of cardiovascular diseases.


Problems in prevention of ischaemic heart disease. The relevance is discussed of the presence of risk factors and their combinations for the prevalence of ischaemic heart disease in a male urban population. The increasing number of risk factors leads to an increase both in the prevalence and in the incidence of ischaemic heart disease. Hypercholesterolaemia, elevated blood pressure, and tobacco smoking have to be considered the most relevant risk factors. Simultaneous presence of 2-3 of these factors was detected in 36% of the men examined. The authors point out the importance of starting a preventive intervention programme in the Czechoslovak population.