Nonallelic histone gene clusters of individual sea urchins (Lytechinus pictus): mapping of homologies in coding and spacer DNA. The linear arrangement and lengths of the spacers and coding regions in the two nonallelic histone gene variant clusters of L. pictus are remarkably homologous by R loop analysis and are similar in general topography to the histone gene repeat units of other sea urchins examined to date. No interventing sequences were detected. The coding regions of these two histone gene variants share considerable sequence homology; however, there are areas of nonhomology in every spacer region and the lengths of the nonhomologous spacers between the H2A and H1 genes are not the same for the two repeat unit classes (inter-gene heterogeneity). Combining length measurements obtained with both R loops and heteroduplexes suggests that the DNA sequences of the analogous leader regions for the two H1 mRNAs are nonhomologous. Similar observations were made for the H4 leader sequences, as well as the trailer region on H2B. S. purpuratus spacer DNA segments share little sequence homology with L. pictus; however, the analgous coding (and possibly flanking) regions have conserved their sequences. The various coding and spacer regions within a repeat unit do not share DNA sequences. Thus certain areas in the sea urchin histone gene repeat units have been highly conserved during evolution, while other areas have been allowed to undergo considerable sequence change not only between species but within a species.


The evolution and sequence comparison of two recently diverged mouse chromosomal beta--globin genes. We have determined the entire nucleotide sequence of a cloned mouse beta--globinminor gene and compared it to the closely related sequence of the betamajor gene. These two genes differ by nine amino acids and presumably evolved from a common ancestral gene as recently as 50 million years ago. Since these genes are closely linked and coordinately expressed, they provide an especially favorable opportunity to assess selection and mutation as these processes affect genes under similar constraints. We find that evolution has preserved these two genes in two short segments of DNA which include their immediately adjacent flanking regions. These regions presumably encode functions that are necessary for proper globin gene expression. In contrast, the more distal flanking sequences and major segments of the long intervening sequences have diverged much more sharply. The homology pattern in these genes also provides considerable insight into the mechanisms by which less constrained nucleotide sequences diverge rapidly. Change in such regions apparently occurs less by point mutation than by insertion, deletion and duplication of relatively short segments of the genome.


The complete sequence of a chromosomal mouse alpha--globin gene reveals elements conserved throughout vertebrate evolution. The mammalian alpha- and beta--globin genes are thought to have evolved from a common ancestral sequence by a duplication event that occurred over 500 million years ago. We have now determined the entire nucleotide sequence of a cloned mouse alpha--globin gene, including regions that flank and interrupt the coding sequence, and have compared this sequence with the sequences of the two mouse beta--globin genes (Konkel, Tilghman and Leder, 1978; Konkel, Maizel and Leder, 1979). Like the two beta genes, the alpha gene is interrupted by two intervening sequences at precisely homologous positions, suggesting that these interruptions were present and have been preserved throughout vertebrate evolution. While the alpha and beta genes conserve considerable (approximately 55%) sequence homology in their coding regions, this homology--with certain interesting exceptions--is lost in the highly divergent flanking and intervening sequences. These exceptions are short preserved sequences positioned in such a way that they might encode signals for transcriptional initiation, poly(A) addition and RNA splicing. Furthermore, a comparison of the recently divererged beta genes and the long separate alpha gene allows us to distinguish two clearly different modes of nucleotide sequence change in evolution: a fast mode which is characterized by drastic sequence alterations involving deletions and insertions, and a slow mode which preserves sequence homology to a large extent and involves mainly point mutations.


Patchwork structure of a bovine satellite DNA. According to a previous restriction nuclease analysis, bovine 1.706 satellite DNA (density 1.706 g/cm3 in CsCl) is organized in an unusual structure of superimposed long- and short-range repeats (Streeck and Zachau, 1978). We have now determined the nucleotide sequence of this satellite DNA in both cloned fragments and fragments from the total satellite DNA. Each long-range repeat unit (about 2350 bp) is divided into four segments. Each segment consists of different variants of a basic 23 bp sequence which is itself composed of a dodecanucleotide and a related undecanucleotide. A total of 2400 nucleotides have been sequenced. Detailed analysis of the sequence divergence reveals that both the overall extent of divergence and the frequency of base changes at individual positions of the 23 bp repeats are characteristically different in the various segments. Preferentially methylated sites and a high incidence of symmetry elements are found. In two of the four segments, 22 of 23 bp of the prototype sequence are included in six overlapping elements of dyad symmetry and in a palindrome. A scheme for the evolution of the satellite DNA from a basic dodecanucleotide is proposed which is based on the different degrees of divergence for the various repeats superimposed in this satellite DNA.


Nonrandom chromosome changes involving the Ig gene-carrying chromosomes 12 and 6 in pristane-induced mouse plasmacytomas. The karyotypes of pristane-induced mouse plasmacytomas were studied by G banding. Only primary tumors or early passage generations were analyzed. In contrast to murine T cell leukemias that showed a regular trisomy of chromosome 15, all plasmacytomas showed a consistent translocation of the distal part of chromosome 15 to either chromosome 6 [rcpT(6;15)] or 12 [T(12;15)]. The specific breakpoints were at 6C, 15D3/E ro D2/3 and 12F2. Early passage generations often showed a mixed population with two different translocations, suggesting polyclonal origin. Considered together with the known karyotypic features of murine and human lymphomas, these findings support the theory that the nonrandom chromosomal changes in lymphoproliferative malignancies are associated with the type of the target cell, rather than with the etiological agent. Moreover, the involvement of the chromosomes known to carry the heavy chain (12) and the light chain (6) determinants, respectively, raises the question of whether the translocations may be related to the DNA level rearrangements known to occur during the differentiation of normal plasma cells.


A study of protein phosphorylation in shape change and Ca++-dependent serotonin release by blood platelets. Upon treatment with agents such as thrombin, collagen or concanavalin A, blood platelets change shape, secrete serotonin and phosphorylate two proteins having molecular weights of approximately 20,000 and 40,000. We have analyzed the relationship of this protein phosphorylation to shape change and release aided by the fact that while shape change occurs independently of extracellular calcium, release of serotonin displays a rather strict calcium requirement. Under limited calcium conditions, where virtually no serotonin release occurs, (Con A)-stimulated phosphorylation is uninhibited. Divalent cations (Mg++, Co++ and Zn++) also inhibit release but not phosphorylation. The microtubule effectors colchicine and D2O show concomitant effects on release and phosphorylation, indicating a microtubule involvement prior to phosphorylation. Papaverine inhibits release and phosphorylation while not strongly influencing shape change, suggesting that shape change does not require phosphorylation. We therefore conclude that phosphorylation of these proteins takes place after shape change but prior to release, and although it may be required for secretion to occur, the two processes are easily separated. Thus phosphorylation of these proteins is not likely to be an integral component of the release mechanism.


A phase relationship associates tRNA structural gene sequences with nucleosome cores. DNA (760 bp) isolated from nucleosome tetramers of staphylococcal nuclease-digested chicken embryo chromatin was highly enriched for tRNA genes and subsequently cloned in E. coli chi 1776. The location of genes coding for chicken embryo tRNALys, tRNAPhe and tRNAiMet within the cloned nucleosome tetramer DNA was determined using restriction endonucleases for which single cleavage sites could be predicted from the respective tRNA base sequence. All our tRNA genes reside nonrandomly at four locations on nucleosome tetramer DNA. The spacing between the tRNA gene locations is approximately 190 bp, similar to the DNA repeat length of chicken embryo chromatin. The four tRNA gene locations were also defined in noncloned nucleosome tetramer DNA highly enriched for tRNA genes. The majority of genes coding for tRNALys, tRNAPhe and tRNAiMet, respectively, are located in equal proportion 40-45, 230, 420 and 610 bp distant from the 5' end of the tRNA-identical strand. Thus the tRNA structural gene sequences all appear to begin about 20 bp "inside" the nucleosome core. As observed with nucleosomal DNA not enriched for tRNA genes, the phase relationship between tRNA genes and nucleosome location is maintained over a distance of 4-6 subsequent nucleosomes. A cloned molecule of nucleosomal DNA containing both a tRNALys gene and a tRNAiMet gene in the same polarity reveals that a phase adjustment might be necessary for the nucleosomes between these two tRNA genes in chicken embryo chromatin.


The primary transcription product of a silkworm alanine tRNA gene: identification of in vitro sites of initiation, termination and processing. A 13.5 Kb fragment of Bombyx mori DNA containing a single tRNA2Ala gene has been cloned, and transcribed in vitro with Xenopus germinal vesicle extracts. The primary transcription product of the tRNA2Ala gene has been isolated and shown to possess an unprocessed triphosphorylated 5' terminus. Products resulting from processing of this transcript have also been isolated and characterized. Complete nucleotide sequence analysis of this cloned alanine tRNA gene and its primary transcript shows that transcription initiates three nucleotides away from the mature tRNA2Ala 5' end and terminates in a U cluster 22 nucleotides beyond the last encoded 3' nucleotide of the mature species. Sequence determination of the products of in vitro maturation shows that in contrast to the tRNA processing mechanism characteristic of procaryotes, the extra 3'-nucleotides in this silkworm tRNA precursor are removed by a single endonucleolytic cleavage.


Isolation of yeast histone genes H2A and H2B. Analysis of cloned sequences for yeast histone genes H2A and H2B reveals that there are only two copies of this pair of genes within the haploid yeast genome. Within each copy, the genes for H2A and H2B are separated by approximately 700 bp of spacer DNA. The two copies are separated from one another in the yeast genome by a minimum distance of 35-60 kb. Sequence homology between the two copies is restricted to the genes for H2A and H2B; the spacer DNA between the genes is nonhomologous. In both copies, the genes for H2A and H2B are divergently transcribed. In addition, both plasmids code for other nonhistone proteins. Sequences coding for histones H3 and H4 have not been detected in the immediate vicinity of the genes for H2A and H2B.


The linkage arrangement of four rabbit beta-like globin genes. Four different regions of rabbit beta-like globin gene sequences designated beta 1, beta 2, beta 3 and beta 4 were identified in a set of clones isolated from a bacteriophage lambda library of chromosomal DNA fragments (Maniatis et al., 1978). Restriction mapping and blot hybridization (Southern, 1975) studies indicate that a subset of these clones containing beta 1 and beta 2 hybridizes to an adult beta-globin cDNA clone (Maniatis et al., 1976) more efficiently than to a human gamma-globin cDNA clone (Wilson et al., 1978), while another subset containing beta 3 and beta 4 displays the converse hybridization specificity. beta 1 was identified as the adult beta-globin gene, while beta 2, beta 3 and beta 4 have not been identified with any known rabbit globin polypeptides. Cross-hybridization and transcriptional orientation experiments indicate that the set of beta-like gene clones contains overlapping restriction fragments encompassing 44 kb of rabbit chromosomal DNA. In addition, all four genes have the same transcriptional orientation and are arranged in the order 5'-beta 4-beta 3-beta 2-beta 1-3'.


The structure and transcription of four linked rabbit beta-like globin genes. Rabbit chromosomal DNA contains a cluster of four linked beta-like globin genes arranged in the orientation 5'-beta 4-(8kb)-beta 3-(5 kb)-beta 2-(7-kb)-beta 1-3'. Determination of the nucleotide sequence of gene beta 1 confirms that this gene corresponds to the second type of two common co-dominant alleles encoding the adult beta-globin chain. With the exception of two nucleotide substitutions in the large intervening sequence (intron), the intron and flanking sequences are identical with the nucleotide sequence of the first type determined by Weissmann et al. (1979). A 14S polyadenylated transcript containing large intron sequences (possibly a mRNA precursor) is detected in the bone marrow cells of anemic rabbits. Gene beta 2 has limited sequence homology to adult and embryonic beta-globin probes and lacks a detectable mRNA transcript in the erythropoietic tissues examined. It contains at least one intervening sequence analogous to the large intron in gene beta 1. Genes beta 3 and beta 4 both contain an intron of 0.8 kb. Partial DNA sequence analysis indicates that the large intron in beta 4 is located between codons for amino acids lysine and leucine in an analogous position to that of the large intron in beta 1. In addition, a second smaller intron interrupts the 5' coding sequences of gene beta 4. Both genes beta 3 and beta 4 are transcribed in embryonic globin-producing cells. Their DNA sequence homology is limited, however, to a segment of approximately 0.2 kb located on the 5' side of the large intron.


Use of a cDNA library for studies on evolution and developmental expression of the chorion multigene families. A cDNA library has been constructed from an RNA preparation highly enriched in silkmoth chorion mRNAs. Many distinct clones have been identified from this library using a stepwise procedure: scoring for infrequent hexanucleotide restriction enzyme recognition sequences; detailed characterization with restriction enzymes that recognize relatively frequent tetranucleotide sequences; probing the arrangement of the corresponding sequences in chromosomal DNA by the Southern procedure; and detailed cross-hybridization analysis. Unique clones, as well as two classes of distinct but related clones, were revealed by hybridization. The cross-hybridization analysis was greatly facilitated by a newly developed, semiquantitative dot hybridization procedure. The same procedure made it feasible to conveniently estimate the relative abundance of several different sequences in an mRNA mixture. Cloned sequences which scored as relatively abundant in total chorion mRNA were tested with stage-specific chorion mRNA at a very stringent criterion of hybridization. They were thus characterized as early, middle or late sequences with respect to development. The characterized cDNA clones can now be used as probes for studying the evolution, chromosomal organization and regulated developmental expression of the chorion multigene families.


Evolution of two major chorion multigene families as inferred from cloned cDNA and protein sequences. Complete or partial sequences are reported from six chorion cDNA clones of the silkmoth Antheraea polyphemus. The proteins encoded belong to the two major chorion protein classes, A and B, each of which is encoded by a multigene family. The sequence comparisons define some major features of the families and suggest how these genes may be evolving. Deletions and insertions might be involved in expanding or contracting internally repetitive regions. Sequence divergence is localized, thus defining sequence domains of distinct evolutionary properties and presumably distinct functions.


Complex population of nonpolyadenylated messenger RNA in mouse brain. The complexity of nonadenylated mRNA [poly(A)-mRNA] has been determined by hybridization with single-copy DNA (scDNA) and cDNA. Our results show that poly(A)- and poly(A)+ mRNA are essentially nonoverlapping (nonhomologous) sequence populations of similar complexity. The sum of the complexities of poly(A)+ mRNA and poly(A)- mRNA is equal to that of total polysomal RNA or total mRNA, or the equivalent of approximately 1.7 x 10(5) different sequences 1.5 kb in length. Poly(A)- mRNA, isolated from polysomal RNA by benzoylated cellulose chromatography, hybridized with 3.6% of the scDNA, corresponding to a complexity of 7.8 x 10(4) different 1.5 kb sequences. The equivalent of only one adenosine tract of approximately 20 nucleotides per 100 poly(A)- mRNA molecules 1.5 kb in size was observed by hybridization with poly(U). cDNA was transcribed from poly(A)- mRNA using random oligonucleotides as primers. Only 1-2% of the single-copy fraction of this cDNA was hybridized using poly(A)+ mRNA as a driver. These results show that poly(A)- mRNA shares few sequences with poly(A)+ mRNA and thus constitutes a separate, complex class of messenger RNA. These measurements preclude the presence of a complex class of bimorphic mRNAs [that is, species present in both poly(A)+ and poly(A)- forms] in brain polysomes.


A mutation which alters initiation of transcription by RNA polymerase III on the Ad5 chromosome. Mutant dl 309 is a viable Ad5 deletion mutant. Whereas wild-type Ad5-infected HeLa cells contain two VAI RNA species [VAI(A) and VAI(G)] which differ by three nucleotides at their 5' ends, dl 309-infected HeLa cells contain VAI(G) but no VAI(A) RNA. Nucleotide sequence analysis indicates that dl 309 lacks two base pairs which precede the 5' end of VAI(A) by 22 nucleotides. Since the 5' ends of VAI RNAs are not processed, the 309 deletion serves to identify a portion of the sequence required for RNA polymerase III initiation. Since dl 309 grows as well as wild-type Ad5 in HeLa cells, the VAI(A) species is not essential for viral growth in these cells.


International activities of the Division of Cancer Treatment, National Cancer Institute. International collaborative activities of the Division of Cancer Treatment, National Cancer Institute, U.S.A., have a long-standing history, and in recent years have expanded to such an extent that an Office of International Treatment Research has been created. The scope of these activities encompasses three areas: 1) participation in International Collaborative Agreements with official national research entities, 2)direct NCI-supported projects, and 3) miscellaneous, less formalized programs. These are described and specific recent contributions are summarized. Emphasis is placed on the importance of international collaboration and exchange of information in searching for new drugs and in maximizing progress in cancer treatment.


The dilemma of estrogen receptors and the response rate to cytotoxic chemotherapy. A problem of comparability analysis. The question of whether estrogen receptor assay predicts for chemotherapy response and, if so, how, is currently controversial. Two papers have recently appeared with widely differing results. Both papers use retrospective analysis and are so heterogeneous as regards assay criteria for positivity, prognostic variable and drugs used that they cannot in truth be meaningfully compared. Resolution of the question will probably have to await prospectively designed trials.


Measuring the quality of medical care: a beginning. Quality of medical care is fundamental; however, the recognition of quality is a complex task. The Patterns of Care Study is examining radiotherapeutic care in cancer throughout the United States, using a new methodology which can be applied to other medical disciplines. The system is described and illustrated as applied to the management of carcinoma of the cervix. Considerable variations in treatment have been identified. These variations can be studied and, where necessary, modified by appropriate educational programs; thus, the quality of medical care can be improved.


[A systematic approach to the study of violence in hockey]. All across Canada, numerous studies every year are being terminated prematurely or simply stored without recognition. Perceived non-significance and a silent and timid approach are among the factors which contribute to the lack of attention to the author's conclusions and recommendations. Therefore the "Comité d'étude sur la violence au hockey amateur au Québec" (CEVH) decided upon a systematic approach to get the interest of specialists and the general public during the entire duration of the investigation. As such, the recommendations were supported by scientific data as well as by public opinion. This approach (POLITICAL TACTIC) resulted in the implementation of more than 80% of the 78 recommendations in less than 6 months after the presentation of the Report. This very season in Quebec amateur hockey, fundamental changes occured.


The role and allergenic importance of storage mites in house dust and other environments. Skin tests were performed on 210 patients with house dust allergy and bronchial asthma or perennial rhinitis using extracts of Dermatophagoides pteronyssinus and of four storage mites most commonly found in house dust in the United Kingdom--Acarus siro, Tyrophagus putrescentiae, Lepidoglyphus (formerly Glycyphagus) destructor and Glycyphagus domesticus. The results of the skin tests were related to certain occupations and living conditions of the patients which could have exposed them to storage mites and some patients were included because their environment seemed especially likely to expose them to these species in order to assess the importance of these conditions. D. pteronyssinus was the most potent of the mite allergens and provoked the largest number of positive tests but a proportion of the storage mite species gave skin reactions which were larger or as large as those of D. pteronyssinus. No significant statistical correlation was found between reactions to D. pteronyssinus and any storage mite but highly significant correlations were found between some storage species. The frequency and strength of reactions to these species were unexpectedly high in view of their irregular occurrence and relative scarcity in house dust. It is suggested that sensitisation to these species occurs through exposure either to localised sources of infestation overlook during the random collection of floor or bedding dust or to infested materials encountered at work or other activities or to infested food or bedding of certain domestic pets. It is concluded that allergy to storage mites is more important and widespread that hitherto realised and is a considerable occupational hazard in farming communities and to those in occupations handling infested materials. Storage mites may also be important allergens for those living in very damp houses where the growth of moulds may encourage the development of Glycyphagus domesticus or other mites.


Comparison of histamine bronchial challenges with the Wright nebulizer and the dosimeter. In twenty adult asthmatics studied in a stable state, histamine bronchial challenges were carried out at one-week intervals with the Wright nebulizer and with the dosimeter-DeVilbiss apparatus. Dose-response curves were analysed for sensitivity, defined as the minimal histamine dose causing a 20% fall in FEV1 (PC20), and for reactivity, defined as the slope of the dose-response curve once the reaction starts to occur. A significant relationship (r=0.80) was found for the sensitivity obtained with the two nebulizers. PC20 was indeed reproducible within a two folds concentration of histamine in sixteen of the twenty patients. A significant difference (P less than 0.01) was found in the reactivity to the two different apparatus, reactivity being greater with the Wright nebulizer in twelve out of fifteen patients so tested. Sensitivity and reactivity showed a borderline relationship (r=0.47) with the Wright nebulizer but not with the dosimeter. A significant correlation (P less than 0.05) was found to exist between the initial FEV1 (in % of the predicted value) and the observed sensitivity and reactivity as assessed with the Wright nebulizer, but not as assessed with the dosimeter. We conclude that histamine bronchial challenges with the Wright nebulizer and with the dosimeter yield reproducible results if the threshold of a 20% fall in FEV1 is taken as positive. However, the two methods produce different results in terms of reactivity.


Acute bronchoconstriction caused by Beclovent and not Vanceril. A 24-year-old asthmatic patient reported acute wheezing immediately after using an aerosol of beclomethasone dipropionate (Beclovent). She was hospitalised for bronchial challenges. Immediate bronchoconstriction was shown after two puffs of this aerosol and after inhalation of the placebo containing all the products of the aerosol less the beclomethasone. Another brand of beclomethasone aerosol, Vanceril, did not produce any reaction. Since the contents of these two beclomethasone aerosols are similar, the authors suggest that rubber or plastic derivative(s) present in the metering valve might have been responsible for the reaction.


Activity units for allergen extracts.

Activity units for allergen extracts. The two present systems of units for the strength of allergen extracts, Noon and p.n.u. are both unsatisfactory. A new activity unit, based on skin prick in humans, is described, and its relationship to in vitro tests is explored. Potency limits for provocation materials used for skin prick tests are suggested.


IgG4: a possible mediator of anaphylaxis in a haemophiliac patient. Immunological investigations have been carried out on a haemophiliac patient who had developed severe anaphylaxis to bovine factor VII administration. Results have shown that the development of anaphylaxis in this patient was associated with a sudden increase in total and specific IgG4 antibodies. The significance of this observation is discussed in relation to current evidence suggesting the involvement of IgG4 antibodies in some anaphylactic conditions.


An examination of the role of vitamin E in glucose-6-phosphate dehydrogenase deficiency. The effect of vitamin E on erythrocyte glutathione stability was studied both in vitro and in vivo on subjects with glucose-6-phosphate dehydrogenase deficiency. The results suggest that lipid membrane peroxidation in glucose-6-phosphate dehydrogenase deficient erythrocytes, which has been postulated to occur under conditions of oxidative stress, does not result in significant depletion of erythrocyte reduced glutathione pools. Vitamin E supplementation in these individuals was shown to have little or no effect on the response of their erythrocytes to oxidative stress.


Methemoglobin reductase: inactivation and protection against inactivation during disc gel electrophoresis. Methemoglobin reductase was found to be inactivated during electrophoresis on ammonium persulfate polymerized polyacrylamide gels. Hemoglobin in various states of ligation offers protection from inactivation. Thiols such as dithiothreitol also offer protection but the effect due to hemoglobin is not provided by its sulfhydryl groups.


Laboratory diagnosis of HB CC-alpha-thalassemia. Routine laboratory investigations of hemoglobinopathies include Hb electrophoresis for abnormal hemoglobins, determination of Hb A2 (alpha 2 delta 2) for beta-thalassemia traits, staining for Hb H (beta 4) inclusions for alpha-thalassemia traits and estimation of Hb F (alpha 2 lambda 2) for the presence of hereditary persistence of fetal hemoglobin genes (HPFH). Frequently, analytical column chromatography and alpha/beta hemoglobin chain synthesis are used in the studies of more complicated hemoglobinopathies. This communication outlines the procedures used in this laboratory for the diagnosis of a case of Hb CC-alpha-thalassemia.


Evaluation of the Beckman Creatinine Analyzer 2. The Beckman Creatinine Analyzer 2 was evaluated in our laboratory. This new instrument determines creatinine by a Jaffe rate method and utilizes an optical detection system which measures the rate of change in absorbance at 520 nm 25.6 seconds after sample introduction. The linearity of the instrument was verified up to creatinine concentrations of 24 mg/dl. Recovery averaged 92% from serum and 97% from urine. Comparison studies using serum specimens were made against a manual, non-kinetic Jaffe method and a Technicon SMA 12/60. Urine comparisons were made against the manual method. All comparisons indicate the Beckman analyzer yields lower creatinine values due to the minimization of interferences from non-creatinine chromogens. No significant change in these correlations was observed with elevated bilirubin concentrations (greater than 10 mg/dl) or with lipemic specimens (triglycerides up to 3390 mg/dl). Both within-day and day-to-day precision were shown to be within the stated manufacturer's specifications for both sera and urine specimens.


Review of Metabolic Screening Program of Children's Hospital, Vancouver, British Columbia. 1971--1977. 1. Between 1971 to 1977, 74,521 urines, collected on filter paper and mailed in, were screened by the Metabolic Screening Program of the Children's Hospital. These represented 45.9% of live births in B.C. hospitals were the program has been available. The mean age of the infants was 4.4 weeks. Urines were examined by chromatography with ethyl acetate-pyridine-water for sugars. 1423 (2.13%) had an abnormal pattern necessitating a repeat urine card. A persistent abnormality was noted in 167 (0.22%) and from these a liquid urine sample was obtained for two dimensional amino acid chromatography and/or a repeat sugar chromatography. 2. In 47 (0.06%) of these a definite metabolic abnormality was confirmed. These included cases of Iminoglycinuria (8), Hartnup trait (4), Nonketotic hyperglycinemia (2), Histidinemia (1), Cystathioninuria (5), Argininosuccinic aciduria (1), Maple Syrup Urine Disease (1), Diabetes Mellitus (1), Renal glycosuria (1) and Persistent galactosuria (3). 201 infants had a slight increase of cystine and/or lysine, and 19 of these were documented to be heterozygous for cystinuria.


Observed range of assay values for plasma and cerebrospinal fluid amino acid levels in infants and children aged 3 months to 10 years. From data collected at the Children's Hospital, Vancouver, during a two year period, 1976 - 1977, we have determined a range of values for children aged 3 months to 6 years for 24 plasma amino acids and for children aged 3 months to 10 years for 18 cerebrospinal fluid amino acids. A total of 26 plasma and 21 cerebrospinal fluid samples were accepted for our reference population. Six of the children in the plasma group were also in the cerebrospinal fluid group, making a total of 39 children in the study.


Assay of erythrocyte glucose-6-phosphate dehydrogenase on the GEMSAEC analyser. A procedure for the assay of glucose-6-phosphate dehydrogenase (EC 1.1.1.49) in erythrocytes on the GEMSAEC centrifugal analyzer using a modification of Beutler's procedure is described. Glucose-6-phosphate dehydrogenase activity is corrected for the contributing activity of 6-phosphogluconate dehydrogenase (EC 1.1.1.44) by using a two-cuvet system in which the activity of the sample incubated with 6-phosphogluconic acid is subtracted from the activity produced in the presence of the combined substrates, glucose-6-phosphate and 6-phosphogluconic acid. An overlay program on the GEMSAEC computer for calculation of results, and the use of a yeast glucose-6-phosphate dehydrogenase control which is stable frozen in dilute solution are discussed.


The qualitative analysis of renal calculi with the polarising microscope. The crystallographic method of Prien and Frondel for the qualitative analysis of renal calculi has been employed for seventeen years. The method was supplemented by occasional chemical tests. Advantages of the crystallographic method over other procedures are the ability to identify specific minerals even in trace amounts, the capability to analyse very small stones, and the rapidity with which analyses can be performed. Data are presented for 1653 calculi.


Sensitive in vivo assay of the phenylalanine hydroxylating system with a small intravenous dose of heptadeutero L-phenylalanine using high pressure liquid chromatography and capillary gas chromatography/mass fragmentography. A method is described which allows the in vivo determination of the phenylalanine hydroxylating system in atypical and classical phenylketonuria. Phenylalanine-d7 is administered i.v. (0.030 g/kg body weight) within 10 min. Tyrosine-d6 in plasma is measured from 30 to 240 min post load by using a computerized capillary gas chromatography/mass fragmentography system. In two patients with hyperphenylalaninemia, the residual activity of the phenylalanine hydroxylating system was 15.7 and 3.7% of the normal, in two phenylketonurics 1.5 and 0.3% respectively. The in vivo figures correspond well to the in vitro assay of the residual activity of the phenylalanine hydroxylase in needle liver biopsy material.


The role of common urinary constituents in the precipitation of ammonium acid urate. A high proportion of the inhibitory activity shown by urine toward precipitation of ammonium acid urate is ultrafilterable and most of this can be accounted for by the common, low molecular weight components of urine. The individual inhibitory effects of sodium chloride, sodium sulphate, potassium chloride, calcium chloride, magnesium sulphate, sodium dihydrogen phosphate, sodium pyrophosphate, citric acid, hippuric acid, creatinine and urea upon the precipitation of ammonium acid urate have been quantified in an aqueous test system.


Halothane interference with pO2 measurements and a method of inhibiting its effects. The oxygen electrode of a blood-gas analyser (Instrumentation Laboratories Inc. Model 313) responded to halothane, the peak of the response being reached some 30-90 min after exposure. The effect was reduced about ten-fold by placing a silicone-rubber membrane over the polypropylene membrane of the oxygen electrode.


Identification of a collagen-like antigen different from C1Q component of complement in serum. The problem of the proportions of hydroxyproline proteins in serum linked to collagen and to the C1Q component of complement was examined. The C1Q component from human and bovine serum, and acid soluble calf skin collagen were purified. Antibodies to soluble collagen were prepared. With these antibodies we found by immunoelectrophoresis that in serum two distinct antigens exist: one reacts with antiserum to C1Q and the other reacts with antiserum to collagen. The collagen-like antigen may be precipitated by sodium chloride 4.3 mol/1.


Quantitation of organic acids in amniotic fluid by gas chromatography. The concentrations of 13 major organic acids in normal human amniotic fluid have been determined by gas chromatography. Each acid showed a large range and an irregular frequency distribution.


An easy procedure for determination of molar activity (or specific activity) of bile acids in bile. An easy procedure is described for determination of the molar activity of the major bile acids, labelled with 14C, in bile. The procedure involves initial enzymatic hydrolysis, by which the amino acid moieties are removed from the glycine and taurine conjugated bile acids by means of choloylglycine hydrolase, followed by thin-layer chromatographic separation of the unconjugated bile acids, cholic acid, chenodeoxycholic acid, and deoxycholic acid. Then the bile acids are eluted from the individual silica gel spots concerned. Finally, determination of the radioactivity by liquid scintillation counting and of the amount of substance by an enzymatic method using 3 alpha-hydroxysteroid dehydrogenase is performed in the eluates. The method requires only a small volume of sample and allows of separate determination of the molar activity of cholic acid and chenodeoxycholic acid, labelled with the same isotope, in the same sample.


Potential methodological errors in the application of equilibrium dialysis to the determination of "free" and "bound" plasma tryptophan. The application of equilibrium dialysis to the determination of the "free", i.e. non-albumin bound fraction of human plasma or serum tryptophan has been re-examined. A potentially serious error in the application of this method is outlined and the necessary correction and some precautionary measures are discussed.


Plasma normetanephrine measurements for detection of pheochromocytoma in patients with hypertension. We describe a simple and specific radioenzymatic assay for measurement of total plasma normetanephrine (NMN), which is an extension of a previously developed procedure for measuring of urinary NMN. Plasma NMN is deconjugated by acid hydrolysis at pH 1.0 and boiled for 20 min. The assay is based on the conversion of NMN to its N-methylated, tritiated derivative metanephrine (3H-MN), utilizing phenylethanolamine N-methyltransferase and S-adenosyl-[3H]methionine. The assay is rapid, sensitive and results can be obtained in less than 4 h. Many antihypertensive drugs tested did not interfere with the assay. This assay could be used for detection of pheochromocytomas in patients with hypertension.


Modification of the normal postural changes in plasma renin activity by the application of positive pressure to the legs. In six normal male subjects plasma renin activity (PRA) rose progressively on standing over a 2 hour period. PRA fell to basal levels after a further 2 hours of recumbency. The rate of rise of PRA on standing was largely abolised following the application of an anti-gravity ('G') suit. Continuous monitoring of blood pressure (BP) heart rate (HR) and packed cell volume (PCV) suggest that it is the fall in effective blood volume which elicits the stimuli to renin release during changes in posture.


Seminal plasma levels of testosterone and 5 alpha-dihydrotestosterone in azoospermic patients. Testosterone (T) and 5 alpha-dihydrotestosterone (DHT) have been measured in seminal plasma of sixteen azoospermic patients with tubular damage (germinal cell arrest or Sertoli cell only syndrome) and five patients affected by azoospermia of obstructive origin. In both groups T values were not significantly different from controls, while DHT was significantly lower in patients affected by azoospermia of obstructive origin.


In vitro biosynthesis of 18-hydroxy-11-deoxycorticosterone from deoxycorticosterone by human adrenal glands removed from patients with hypercorticism. The biosynthesis of 18-hydroxy-11-deoxycorticosterone (18-OH-DOC) was studied by in vitro incubations of human adrenals resected from patients with Cushing's syndrome, using deoxycorticosterone as precursor. 18-OH-DOC synthesis was similar in adrenals from normal subjects and in those derived from patients with different types of Cushing's syndrome (hyperplasia, adenoma, carcinoma). The in vitro addition of o,p'-DDD (5 x 10(-3) M) to an adrenocortical carcinoma incubation did not alter 18-OH-DOC biosynthesis. By contrast, treatment of the patient with o,p'-DDD prior to adrenalectomy resulted in a low production of 18-OH-DOC in vitro but only when cortisol synthesis was drastically decreased.


Ectopic pituitary adenomas.

Ectopic pituitary adenomas. We report here four cases representing different forms of ectopic pituitary adenomas localized: (1) in the nasal cavity, (2) in the sphenodial sinus, (3) on the sphenoidal wing and (4) on the petrous temporal bone. Ectopic pituitary adenomas may probably occur in extrasellar pituitary tissue deposited along the route of fetal development or as a result of intracranial dissemination of adenomatous cells during operation. It is assumed that in some cases ectopic pituitary tissue may retain endocrine function leading to a sustained pituitary function even after a radical hypophysectomy.


Metoclopramide induced growth hormone release in hypogonadal males. The growth hormone (GH) response to 10 mg intravenous metoclopramide (MCP) ('Maxolon'--Beecham Laboratories) was determined in ten hypogonadal adult males. All the subjects responded with a peak growth hormone level greater than 14 mU/l within 1 hour of administration of MCP. Seven normal adult males did not respond to MCP administration. It is concluded that MCP stimulation may be a useful test for both prolactin and growth hormone reserve in hypogonadal males.


Effects of 1,25-dihydroxycholecalciferol on 47calcium absorption in post-menopausal osteoporosis. Measurement of 47Calcium absorption was performed on eleven women with post-menopausal osteoporosis. The study was repeated after 10 days treatment with 1 microgram daily of 1,25(OH)2D3. These patients showed a statistically significant improvement of fractional calcium absorption that was inversely correlated to the basal values. The prompt improvement of the intestinal calcium transport in post-menopausal osteoporotic women, a few days after the administration of physiological doses of 1,25(OH)2D3, suggests that these patients synthesize inappropriately small amounts of 1,25(OH)2D3 because of their oestrogen deficiency. This could be an important pathogenetic factor in post-menopausal osteoporosis, as the efficiency of the adaptation of calcium absorption to low calcium intakes is dependent on 1,25(OH)2D3.


Immunoreactive somatomedin B in acromegaly and in Turner's syndrome. Serum somatomedin B was measured by radioimmunoassay in forty-seven normal subjects, twenty-nine patients with acromegaly before and twenty-four after treatment, and eighteen patients with Turner's syndrome. Somatomedin B levels were significantly elevated in untreated acromegaly and in Turner's syndrome compared with the control group; they decreased following treatment of acromegaly. Because of the overlap between the groups, little information could be obtained from single somatomedin B estimations, which could, therefore, not replace dynamic tests of growth hormone secretion. No correlation between growth hormone and somatomedin B in acromegaly was detected; however, somatomedin B appeared to be related to the insulin response during the oral glucose tolerance test. In Turner's syndrome, no relationship between somatomedin B and insulin production, urinary oestrogen excretion, growth hormone secretion, gonadotrophin levels, age or height was found. The reason for the raised somatomedin B levels in Turner's syndrome remains at present unknown.


Cimetidine induced prolactin (PRL) release in thyroid disease. Effects of acute intravenous administration of cimetidine on serum levels or PRL, TSH, LH and FSH were investigated in six healthy euthyroid, five hypothyroid and five hyperthyroid patients. Cimetidine failed to alter serum TSH levels in the hypothyroid patients. There was no change in LH and FSH levels except in two euthyroid subjects in whom significant rise in LH occurred. However, in all subjects, whether euthyroid, hypo- or hyperthyroid, the serum PRL concentration increased two to three fold above basal levels. The peak levels of serum PRL occurred at 15 min following the drug administration. The response was not blunted in the older age group. We conclude that unlike the TRH-mediated PRL release, the cimetidine-induced PRL release is independent of the levels of thyroid hormone.


Bromocriptine treatment of oligospermia: a double blind study. A double blind controlled study of bromocriptine treatment of oligospermia was carried out. Out of fifty-one men who originally volunteered to the study there were forty who took the drug for 12 weeks as requested. All the partners of these men had failed to conceive, and in each case the pretreatment sperm count had been below 40 million/ml on two or several occasions. The pretreatment serum prolactin concentrations were similar in patients given bromocriptine (N = 20) and placebo (N = 20). There were three men in either group whose pretreatment serum prolactin concentration was in excess of 30 micrograms/l, the highest value being 96 micrograms/l. While bromocriptine effectively decreased the serum prolactin concentration, it had no significant effect over placebo on sperm volume, motility and morphology. In the bromocriptine group, sperm count increased to or above 40 million/ml in five out of twenty men, while in the placebo group this occurred in nine out of twenty patients. The plasma testosterone and dihydrotestosterone levels increased slightly during treatment in both groups, but no significant difference was observed between bromocriptine and placebo treated patients. One wife of a bromocriptine-treated man and two wives of placebo-treated men became pregnant during treatment. In this study bromocriptine was no more effective than placebo in the treatment of oligospermia.


Heterogeneity in adrenal steroidogenesis in normal men and women. Adrenal steroidogenesis has been studied in vivo in normal men and women. Serum levels of nine steroids on the biosynthetic pathway (the delta 5 3-beta-hydroxysteroids, pregnenolone (Pe), 17 alpha-hydroxypregnenolone (17Pe), dehydroepiandrosterone (DHEA), androstenediol (Adiol), and their delta 4 3-keto counterparts, progesterone (Po), 17 alpha-hydroxyprogesterone (17Po), androstenedione (Adione), and testosterone (T)) as well as cortisol were measured during adrenal suppression and stimulation. This study demonstrates a marked heterogeneity in adrenal steroid responses between different subjects in the normal population. Thus, in three subjects ACTH stimulation from a dexamethasone-suppressed state resulted in a far greater increment of 17Po than in the other nineteen normal subjects. These three individuals (designated Type 2 responders) may have a partial deficiency of 21-hydroxylase activity. In the remaining nineteen subjects (designated as Type 1 responders) the women had a greater increment of Adiol (P less than 0.05) and a lower increment of Po (P less than 0.01) than the men, suggesting that adrenal 3-beta-hydroxysteroid dehydrogenase/isomerase activity may be slightly lower in women than men.


Glucagon metabolism in normal subjects and in cirrhotic patients before and after portasystemic venous shunt surgery. The effect of portasystemic shunt surgery on basal immunoreactive glucagon (IRG) levels, metabolic clearance rate (MCR) and t 1/2 for glucagon decay, and basal systemic delivery rate (BSDR) of glucagon was investigated in paired studies in ten cirrhotic subjects. The degree of hepatocellular dysfunction and extent of portasystemic venous shunting was also recorded. Basal IRG levels were highest in the post-shunt (mean +/- SEM, 382 +/- 73 pg/ml) as compared to the pre-shunt (213 +/- 27 pg/ml; P less than 0.05) cirrhotic and control (53 +/- 13 pg/ml; P less than 0.005) groups. The MCR of glucagon was similar in control (13.0 +/- 1.3 ml/kg/min) and pre-shunt cirrhotic patients (13.3 +/- 1.7 ml/kg/min) but was significantly (P less than 0.02) decreased in the post-shunt cirrhotics (7.6 +/- 1.3 ml/kg/min). The t 1/2 for glucagon decay was similar in the control and cirrhotic groups. The BSDR, an estimate of pancreatic A cell secretion, was increased four-fold (P less than 0.01) in the pre-shunt (3042 +/- 454 pg/kg/min) and post-shunt (2518 +/- 535 pg/kg/min) cirrhotic groups, as compared to controls (750 +/- 244 pg/kg/min). It is concluded that (a) in the presence of cirrhosis, the magnitude of portasystemic shunting is important in determining the degree of hyperglucagonaemia; (b) in preshunt cirrhotics raised basal IRG levels are principally due to A cell hypersecretion of glucagon whereas in post-shunt cirrhotics riased IRG levels reflect both A cell hypersecretion and delayed clearance of glucagon; and (c) acute shunting of splanchnic venous blood away from the liver reduces the clearance of glucagon, suggesting that, in man, the liver contributes to the clearance of circulating glucagon.


Twenty-four hour profiles of circulating androgens and oestrogens in male puberty with and without gynaecomastia. The possible mechanisms involved in the development of transient gynaecomastia during male puberty have been investigated by studying 24 h profiles of circulating androstenedione (Ao) and testosterone (T) and their oestrogen pairs oestrone (E1) and oestradiol(E2), in eight boys with simple delayed puberty, eleven boys with pubertal gynaecomastia (three of whom were re-tested after its spontaneous resolution), and two normal adult men. No differences were observed between the 24h T and Ao profiles of pubertal boys with or without gynaecomastia; we confirmed that the initial T rise was nocturnal, associated with sleep. Late in puberty daytime T levels also rise. A small rise in 24 h Ao was seen, but this was not closely related to the stage of puberty. The major new finding was that E2 and to a lesser extent E1 levels are high relative to T for prolonged periods of the afternoon and evening (when T levels are lowest) in male puberty. A frequent finding, seen only in boys with gynaecomastia and one who later developed it, was of elevated and markedly fluctuating levels of plasma E2, and an absolute increase in the area under the 24 h E2 profile and between the E2 and T profiles. These fell towards normal in three boys who were re-tested after resolution of gynaecomastia. In a minority of subjects T and E2 were quite closely correlated, suggesting that in them rapid aromatization of T was occurring within or outside the testis. We conclude that normal male puberty is associated with relative oestrogen dominance particulary in the daytime. In boys with gynaecomastia there is in addition often an absolute elevation of E2 with or without E1, while 24 h T levels are submaximal. Normal men probably require sustained adult circulating T levels to prevent their oestrogens from stimulating breast development.


Linkage analysis using heterozygote detection in phenylketonuria. This linkage investigation was undertaken utilizing an improved method for phenylketonuria (PKU) heterozygote detection. This method is based on studies of semi-fasting, noon-time, blood specimens obtained from 85 obligate heterozgotes and 45 controls who were neither pregnant nor on birth control medication. The best separation between heterozygotes and normals was achieved with a discriminant function involving the logarithms of the serum concentrations of phenylalanine, tyrosine and tryptophan. The theoretical overlap area between the distributions of heterozygotes and controls, based on the above function, was between the distributions of heterozygotes and controls, based on the above function, was 3.75%. In 19 obligate heterozygotes and 13 controls who were either pregnant or on birth control medication, the best separation was achieved with a discriminant function involving the logarithms of the serum concentrations of phenylalanine and tyrosine. The theoretical overlap area was 8.23%. These equations identified heterozygotes with sufficient accuracy to permit efficient genetic linkage analysis. We were unable to demonstrate genetic linkage between the PKU locus and 15 common blood, serum, and urinary markers. All but loose linkage (theta greater than 0.3) was excluded for Rh, ABO, Gc, Kidd, and AP. Moderate linkage exclusion (theta less than 0.2) was shown for PGM, Duffy,  Hp, MNS, HL--A, and Kell. Close linkage (theta less than 0.1) was excluded for Amy2, 6PGD, P, and ADA. We were unable to find linkage heterogeneity between the Amish and non-Amish populations.


Familial cardiomyopathy. Autosomally, dominantly inherited congestive cardiomyopathy with two cases of septal hypertrophy in one family. A family with inherited congestive cardiomyopathy is presented. The diagnosis is based on clinical, morphological and laboratory evaluations. The first observed sign of the disease is arrhythmia and/or conduction defects. The onset of symptoms of pump failure is in adult life, and affected persons die within several years. Three persons have died suddenly. Septal hypertrophy was present in two affected persons. The mode of transmission is probably autosomal dominant. The recognition of arrhythmia as an early sign of the disease offers the opportunity of an early diagnosis.


Familial congenital diaphragmatic hernia: prenatal diagnostic approach and analysis of twelve families. Congenital diaphragmatic hernia is generally recognized as a sporadic malformation with little or no risk of recurrence. A family with three affected individuals in two generations is presented. In addition, new prenatal diagnostic techniques including ultrasonography and amniography are discussed. A comparison of associated physical characteristics in isolated versus twelve familial cases of diaphragmatic hernia is presented. In the familial group, there was a higher incidence of affected males (M:F ratio = 2.1 versus 0.67), a higher incidence of bilateral defects (20% versus 3%) and a lower incidence of additional life-threatening malforamtions 3.6% versus 47%). Analysis of available pedigree data favors multifactorial inheritance with a high male: female sex ratio as the most probable mode of transmission.


A reciprocal translocation (X;11) in a female with gonadal dysgenesis. A 24-year-old female patient was referred for evaluation of primary amenorrhea. Endocrine studies showed elevated gonadotropins, consistent with gonadal failure. At laparoscopy, a normal nulligravid uterus, normal fallopian tubes, and bilateral streak gonads were observed. Histologic studies showed that the left gonad consisted entirely of fibrous tissue, confirming the presence of streak gonads. Chromosome banding studies of peripheral blood and cultures of tissue from the left gonad demonstrated a 46,X,rcp(X;11)(q22;q13) karyotype. A review of reports of X-autosome reciprocal translocations indicated that abnormal gonadal development is associated with break-points in the mid-region of the long arm of the X chromosome.


The Aarskog (facio-digito-genital) syndrome. A kindred with the Aarskog Syndrome (facio-digito-genital syndrome) is presented. The favorable outcome of this syndrome in terms of final height, intelligence and life expectancy is emphasized. It is pointed out that cleft lip and cleft pastic dermatoglyphic pattern includes simian creases and distal axial tiriradii. The pedigree analysis from 14 known kindreds strongly suggests an X-linked recessive mode of inheritance. It is noteworthy that the female carriers frequently showed varying and milder expression of the syndrome, especially in their facies and hands.


Retinal manifestations in familial juvenile nephronophthisis. Three families affected by juvenile nephronophthisis are described and genetically analyzed. The extrarenal manifestations of the disease include retinal findings such as congenital Leber amaurosis, retinitis pigmentosa and sector retinitis pigmentosa. Autosomal recessive transmission and the wide phenotypic spectrum are discussed. The variability in the retinal findings is explained by the presence of a pleiotropic gene with variable expressivity.


Healthy female carriers of a gene for the Alport syndrome: importance for genetic counseling. Of 305 female obligatory carriers of a gene for the Alport syndrome identified in the literature, 37 (12.1%) had no clinical or laboratory signs or symptoms (renal, audiometric, ophthalmological) of disease. The prevalence of unaffected female obligatory carriers appeared not to vary according to age or to the severity of the illness in male members of any pedigree. Normal clinical or laboratory data for any female liable for the Alport syndrome must thus be interpreted with caution.


Genetic basis of acquired C4 deficiency. A study of the family of a patient who had an SLE-like syndrome and an extremely low serum C4 revealed an inheritance of C4 types and HLA region markers which indicated that the patient had 60--70% of "normal" C4 level prior to the onset of disease. Thus the extremely low C4 level during her disease may result from a combination of genetically determined low normal C4 and increased consumption/hyposynthesis secondary to her SLE.


An autosomal dominantly inherited syndrome of facial asymmetry, esotropia, amblyopia, and submucous cleft palate (Bencze syndrome). This is the second report of a dominantly inherited syndrome of facial asymmetry, esotropia, and amblyopia (Bencze syndrome). The phenotypic spectrum is expanded to include submucous cleft palate. The observation for the first time of male-to-male transmission seems to confirm an autosomal dominant mode of inheritance. The facial asymmetry in this family was mild and did not require surgical intervention. With the exception of one patient who had other abnormalities, intelligence was normal.


Phenotypic variation in two patients with a ring chromosome 22. Two severely mentally retarded patients with a ring chromosome 22 presented with disparate phenotypes: one patient manifested only minimal dysmorphic features, whereas the other had a distinctive pattern of anomalies consisting of an abnormal skull configuration with mild maxillary hypoplasia, a large nose, thick full lips, a protruding tongue, lymphedema, hypotonia and an unsteady gait. The findings in these and previously reported patients indicate that a ring chromosome 22 is usually associated with moderate to severe mental retardation, with a range of dysplastic features from mild and nonspecific to more marked and distinctive.


Prenatal diagnosis of galactosaemia in six pregnancies -- possible complications with rare alleles of the galactose 1-phosphate uridyl transferase locus. We describe our experience in prenatal diagnosis of six foetuses at risk for galactosaemia. In one family the parents were both shown to be double heterozygotes at the galactose 1-phosphate uridyl transferase (Gal-PUT) locus, the mother having a Duarte/Los Angeles and the father a Duarte/galactosaemia genotype. The foetus (and an older brother previously thought to have classical galactosaemia) was also a Duarte/galactosaemia double heterozygote. In the other five families, the parents and three foetuses were heterozygous carriers of the galactosaemia gene, one of the foetuses had galactosaemia, and one was homozygous for the normal gene. It is concluded that by a combination of family studies and assay of cultured amniotic cell Gal-PUT, accurate prediction of the foetal Gal-PUT genotype is now possible.


Neuraminidase deficiency in the original patient with the Goldberg syndrome. Homogenates of cultured skin fibroblasts from a non-ambulatory, 20-year-old male with cherry-red spots, corneal clouding, seizures, mental retardation, dysostosis multiplex, dwarfism, coarse facies and loss of vision, originally described by Goldberg et al. (1971), have diminished neuraminidase activity and an excess of neuraminic acid-rich compounds. Specifically, these cells have 2-17% normal neuraminidase when measured with 2-(3' methoxyphenyl)-N-acetyl-alpha-neuraminic acid, N-acetyl-neuramin-lactose and fetuin. Activities of 12 other lysosomal enzymes were either at or above the range of normal control fibroblasts. Total neuraminic acid concentration was 44.3 nmol/mg protein versus an average control value of 14.2. It is concluded that the Goldberg syndrome should be considered, along with mucolipidosis I and the cherry-red spot -- myoclonus syndrome, as resulting from a primary neuraminidase deficiency.


Inheritance of microtia and aural atresia in a family with five affected members. Only a few reports of congenital meatal atresia and microtia have been published. Dominant as well as recessive inheritance has been suggested. We report a family with five affected members in two generations. An irregularly dominant transmission seems to be the most likely explanation.


A distinct variant of the Ehlers-Danlos syndrome. Two unrelated males presented a distinct syndrome, consisting mainly of mental retardation, short stature, wrinkled facies, curly and fine hair, scanty eyebrows and eyelashes, telecanthus, periodontitis, hypermobility of the joints, hyperextensibility and fragility of the skin, multiple nevi, papiraceous scars, bruisability, varicose veins, pectus excavatum, winged scapulae, pes planus and bilateral cryptorchidism. Since some features were typical of Ehlers-Danlos Syndrome (EDS), the clinical data were analyzed comparatively with the different types of EDS. The individualization of a distinct variant is concluded. Increased paternal age at the birth of both cases suggests a de novo dominant mutation.


Fanconi's anaemia associated with haemophilia A. Fanconi's anaemia and haemophilia A are born inherited diseases creating haemostatic defects. The association of these two rare diseases in one patient is described. The patient's haemophilia was studied with a newly developed immunological technique determining the plasma antigen associated with Factor VIII activity, and was found to be a genetic variant of moderately severe haemophilia A. It was not possible to demonstrate a common bone marrow defect or a common immunological or genetical background of the two diseases. The double haemostatic defect created, i.e. Factor VIII deficiency and thrombocytopenia, resulted in only a slight increase in bleeding tendency. A favourable result was obtained with corticosteroid and androgenic treatment.


Left atrial type B receptor response during hypothermia in dogs. 1. The response of left atrial type B receptors to hypothermia and its effect on the V-wave pressure-receptor activity relationship was studied in ten anaesthetized and thoracotomized dogs. Hypothermia was produced by surface cooling of the animal. 2. With the drop in the body temperature there was a corresponding fall in the peak frequency of discharge. Reduction in the number of impulses per cardiac cycle and the average resting activity of these receptors became significant only when body temperature fell below 31 degrees C. The Q10 for these receptors is about 2.5, i.e., similar to that for the other mechanoreceptors and consistent with the view that these receptors are in parellel with the muscle fibres. 3. The V-wave pressure-receptor activity relationships were not altered by acute hypothermia.


Properties of inactive renin in human plasma. 1. "Inactive" renin in human plasma can be revealed by pH 3.3- or cold-mediated activation and in normal plasma represented 76% of the "total" renin. 2. Pregnancy plasma contained considerably more "inactive" renin and consisted of 93% of "total" renin. 3. "Active" renin in normal plasma had an apparent molecular weight of 43,000 compared with 60,000 for "active" renin in pregnancy plasma by gel filtration. 4. "Inactive" renin in pregnancy plasma also had an apparent molecular weight of 60,000, while in normal plasma there were two peaks of inactive renin at 62,000 and 46,000. 5. After affinity chromatography of a protein preparation from pregnancy plasma on Concanavalin A-Sepharose activation by pH 3.3 could no longer be produced, suggesting that the activating factor had been removed, as would occur if it were not a glycoprotein. When pepsinogen, in a concentration similar to that found in plasma, was added prior to dialysis to pH 3.3 activation was restored. 6. Ion-exchange chromatography demonstrated that at pH 8.4 "inactive" renin bore slightly less negative charges than "active" renin. 7. "Inactive" renin in human plasma therefore appears to be a larger molecular weight species than the "active" renin in normal plasma and is capable of activation during treatment to pH 3.3 or cold with no apparent alteration in size. The results suggest an important role of pepsin (after conversion from pepsinogen) in the activation of "inactive" renin during dialysis at pH 3.3.


The isolated lung strip and single open tracheal ring: a convenient combination for characterizing Schultz Dale anaphylactic contractions in the peripheral and central airways of the guinea-pig. 1. The use of the isolated lung strip and single open tracheal ring for studying Schultz Dale anaphylactic contractions in both the peripheral and central airways is described. 2. This method is particularly relevant to studies of anaphylaxis because many preparations may be obtained from a single sensitized animal. 3. Isolated preparations from control guinea-pigs did not respond to ovalbumin, whereas the lung strip and trachea taken from guinea-pigs sensitized to ovalbumin, contracted markedly to that antigen. 4. The peripheral and central airways from sensitized animals responded in the same way to antigen challenge.


Dopamine and the enhanced renal response to aldosterone in the rat on a high potassium diet. 1. The enhanced renal response to aldosterone observed in rats on a high potassium diet is reduced by the administration of either haloperidol, a dopamine receptor blocker, or carbidopa, a peripheral blocker of dopa decarboxylase. This suggests that one peripheral action of dopamine is to modulate the action of aldosterone on the kidney. 2. No difference in urinary dopamine excretion after an 18 h fast was found in rats that had been either on a control diet or a high potassium diet. Both groups of rats responded by a similar elevation after an acute potassium load. This suggests that dopaminergic mediation of the enhanced response to aldosterone in rats on a chronic high potassium diet is not detectable by changes in urinary dopamine excretion.


Embryonic and early fetal loss.

Embryonic and early fetal loss. Embryonic and early fetal loss occurs in approximately one out of two pregnancies. Although this rate might appear unreasonably high, it is borne out by several types of studies and further confirmed by careful animal study. It now appears that karyotype deviation is common in early abortuses and probably underlies the high rate of gross abnormality seen in this group. In addition, environmental factors (radiation, smoking, anesthetics, and previous therapeutic abortion) may contribute to this wastage. Study and analysis of the abortus have allowed identification of a number of new embryonic and fetal syndromes. Careful description and classification combined with continued monitoring of abortuses shows promise both as an adjunct to reproductive counseling and as an important defense against teratogenic exposure. It is through this study that the etiology and ultimately the control of preventable pregnancy wastage will be understood and implemented.


Prenatal diagnosis of chromosomal abnormalities and neural tube defects. We have reviewed the current status of prenatal diagnosis for chromosomal indications and for neural tube defects. The number of pregnancies monitored will increase as greater resources become available and as public education about genetics increases. The methodology has proved to be a powerful means of preventing the birth of individuals with significant genetic defects, thereby sparing both parents and society from the burdens produced by such disorders. In the future, it is likely to be even more effective.


Dose-dependent elimination kinetics of theophylline. There are an increasing number of clinical and experimental studies appearing in the literature suggesting that the elimination kinetics of theophylline may be dose-dependent in man. Evidence of nonlinearity includes observations that steady-state serum theophylline concentrations may increase disproportionately with increases in dose and that the elimination half-life of theophylline after multiple doses is longer than after single doses. Theophylline is apparently eliminated by parallel Michaelis-Menten and first-order kinetics. Although no specific studies of theophylline metabolism have been carried out to determine the causes of nonlinearity, it would appear that 1-demethylation of theophylline is a potentially saturable metabolic pathway. Dietary methylxanthines or their metabolites may influence the rate of elimination of theophylline and contribute to the causes of nonlinearity. Until the elimination kinetics of theophylline have been studied in more detail as a function of dose, it would seem prudent to increase doses of theophylline cautiously to minimise the risks of adverse side effects which might be associated with unexpected accumulation of theophylline.


Hypothesis for the individualisation of drug dosage. Computer simulations based on the pharmacokinetics of chloramphenicol and theophylline in patients, indicate a very strong correlation (r = 0.988 for chloramphenicol and r = 0.971 for theophylline) between log maintenance dose required to achieve a desired average drug concentration in serum at steady-state, and the drug concentration in serum 6 hours after an initial test dose administered by constant rate intravenous infusion over 0.5h. Accordingly, we have developed a nomogram to predict individual daily dosing requirements for these drugs in uncomplicated patients from a single serum assay following an initial dose. Within defined limits, predictions made with the nomogram are essentially equivalent to those made by iraditional pharmacokinetic methods which require substantially more drug concentration-time data following a test dose. Predictions based on the nomogram are relatively unaffected by small but typical errors in magnitude of the test dose, infusion time, sampling time and assay. Protocols for the administration of the test dose other than described, e.g. administration of an oral theophylline solution, may be equally useful for dosage predictions. In principle, this approach should apply to other drugs.


Bacterial control in the burn wound.

Bacterial control in the burn wound. The goal in the treatment of septic burn wounds, as in any infection, is to reestablish the normal balance between the bacteria and the host defense. This is readily done by controlling the numerical level of bacteria while expending all efforts to remove irreversibly damaged tissue and obtaining a closed wound by autograft with the patient's own skin as rapidly as possible.


Infections following surgery for head and neck cancer. Cancer of the head and neck area disturbs the balance between the bacterial organisms of the area and the host-defense mechanisms. The very nature of treatment of this cancer predisposes patients to infection. An awareness of the multiplicity of factors that lead to these complications and the understanding of the various defense mechanisms of the host provide the basis for a rational approach to the successful treatment of these patients.


Difficult wounds: radiation wounds.

Difficult wounds: radiation wounds. In an era of modern radiotherapy, problems associated with the indiscriminate treatment of benign disease have largely disappeared. Skin sparing effects of super voltage radiation equipment make the problems previously seen with orthovoltage equipment less frequent. Vigilance on the part of the workers in the field, in general, protects from the disasters that befell Thomas Edison's laboratory assistant. Despite these modern advances, the reconstructive surgeon often faces problems of managing acute local radiation injury from accident following planned therapeutic radiation or the ulcerations and breakdowns seen months or years after radiation therapy. The single most serious hazard to surgery in radiated tissue is the lodgment of bacteria in this tissue rendered avascular by the radiation and secondary necrosis from the infection itself. The principles of management are no different from those used for other chronic granulating wounds: local wound care, appropriate topical antibacterial therapy, systemic antibiotics during the perioperative period and, most importantly, adequate soft tissue coverage.


The use of antimicrobial agents.

The use of antimicrobial agents. Antibiotics are potent preventive and therapeutic agents. However, they do not render humans germ-free. The incidence of infection in most operations has changed little in the decades since antibiotics have been available. These drugs should be used with therapeutic precision with recognition of their limitations and, as pointed out in this article, their dangers. "Shoot with a rifle--not a shotgun."


Maxillofacial infections.

Maxillofacial infections. Maxillofacial infections, either surgical or nonsurgical, are relatively infrequent, probably because of the excellent regional blood supply. Established infections are usually recognized early owing to the obvious signs of infection and the early symptoms. Fortunately, most infections respond to the penicillin group of antibiotics. However, spread of maxillofacial infections may be disastrous, not only locally and regionally but possibly even systemically. There is always a chance of a lethal complication from a maxillofacial infection.


Carbon dioxide equilibration in canine lungs during rebreathing and acute alkalosis. 1. The left lower lobe of the lungs of six anaesthetized dogs were isolated by the introduction of a bronchial cannula at thoracotomy. Catheters were introduced into the main pulmonary artery and a vein draining the isolated lobe. 2. Blood-gas pressures and pH were measured across the isolated lobe and compared with gas pressures in alveolar samples from the lobe. 3. When the isolated lobe was allowed to reach gaseous equilibrium with pulmonary arterial blood for 30 min, there was no significant difference between alveolar and pulmonary venous PCO2. Mean values of whole-blood base excess were similar in pulmonary arterial and pulmonary venous blood. 4. After injection of 20 ml of 8.4% sodium bicarbonate solution into a peripheral vein, PCO2, pH and plasma bicarbonate concentrations rose in the mixed venous blood. There was no change of whole-blood base excess across the lung, indicating that HCO3-, as distinct from dissolved CO2, did not enter lung tissue in measurable amounts. 5. No systematic alveolar-pulmonary venous PCO2 differences were demonstrated in this preparation other than those explicable by maldistribution of lobar blood flow.


Changes in arterial blood gases during and after a period of oxygen breathing in patients with chronic hypercapnic respiratory failure and in patients with asthma. 1. Ten patients with chronic hypercapnic respiratory failure (group 1) and eight patients with asthma (group 2) breathed pure O2 from an MC mask for 60 min. Blood gases were measured during this period and for the subsequent 45 min. 2. In mine of ten patients in group 1 and in all eight patients in group 2 arterial O2 tension (Pa,O2) fell to values lower than had been obtained before O2 was given. 3. These undershoots in Pa,O2 are unrelated to changing CO2 stores or to hypoventilation, and are more likely due to persistence of altered ventilation-perfusion ratios associated with O2 breathing. 4. Magnitude of the undershoots is usually small, and periods of less than 15 min of O2 are unlikely to be harmful.


Effect of isoprenaline on the plasma concentrations of angiotensin III in rats. 1. A new column-chromatographic method is described for the simple and reproducible determination of the concentration of [des-Asp1]angiotensin II (angiotensin III) in rat plasma. 2. The method uses the different abilities of the ion-exchange resins Dowex 1 (X8) and Bio Rex 70 to bind angiotensin II and angiotensin III. Under the conditions used, Bio Rex 70 binds only angiotensin III. Angiotensin II and its hexapeptide metabolite [des-Asp1,des-Arg2]angiotensin II pass the resin with the effluent. Dowex 1 (X8) binds angiotensin II and the hexapeptide metabolite, whereas it does not extract angiotensin III. It does not separate angiotensin II from the hexapeptide. Therefore the sum of both peptides is expressed as angiotensin II-like activity. The ratio of the concentrations, angiotensin II/hexapeptide, was 5:1. 3. In normal rats the plasma concentration of angiotensin III was 20 fmol/ml (SD 15; n = 8), and angiotensin II-like activity was 60 fmol/ml (SD 35; n = 8). 4. The beta-sympathomimetic amine isoprenaline caused a time- and dose-dependent increase in plasma angiotensin III and angiotensin II-like activities. 5. Under the conditions studied angiotensin III contributed approximately 25% to the total amount of angiotensins in plasma.


Dissociation in the excretion of different aldosterone metabolites and unmetabolized ('free') aldosterone in hypertension. 1. The determination of aldosterone-18-glucuronide (pH 1-labile aldosterone) was complemented by concomitant measurements of free urinary aldosterone and tetrahydroaldosterone in 307 patients, most of whom were hypertensive. In 38 cases (12.3%) the normal, aldosterone-18-glucuronide concentration was clinically misleading, but increased free aldosterone and/or tetrahydroaldosterone values suggested the presence of hyperaldosteronism, which in many of these cases was confirmed by elevated excretion of the possible major aldosterone precursor 18-hydroxycorticosterone (18-OH-B). 2. Of 224 patients with essential hypertension and normal or low plasma renin activity 18 had an elevated free aldosterone and/or tetrahydroaldosterone excretion without increased aldosterone-18-glucuronide. These cases may represent early or pre-symptomatic forms of primary hyperaldosteronism. In other cases, particularly when tetrahydroaldosterone was increased alone, abnormalities of aldosterone metabolism were suspected. 3. In two out of 15 patients with primary hyperaldosteronism, aldosterone-18-glucuronide values were frequently found to be normal, although elevations were noted in other variables. However, no relation to the morphological abnormality (adenoma versus hyperplasia) was seen.


Effect of ouabain in therapeutic concentrations on K+ exchange and contraction of human and rabbit myocardium. 1. Experiments were undertakn on strips of human atrial and ventricular muscle, on right ventricular rabbit papillary muscles and on the rabbit interventricular septum. 2. In all preparations an increase of developed tension after 30 min exposure to ouabain was apparent only at or above a concentration of 10(-7) mol/l (73 micrograms/l). 3. In the rabbit septum there was a net loss of potassium on exposure to ouabain if the concentration was greater than 10(-7) mol/l. There was no net loss, or gain, of potassium at lower concentrations. Loss of potassium, indicating inhibition of the sodium pump, could not be dissociated from the positive inotropic effect of ouabain. 4. The greater sensitivity to ouabain of human myocardium in vivo (10(-8) mol/l) than in vitro (10(-7) mol/l) may be due to cellular changes occurring in isolated preparations.


Are mineralocorticoid receptors present in human renal adenocarcinoma? 1. The presence of high-affinity sites for [3H]-aldosterone was shown in the normal human renal tissue. 2. [3H]Aldosterone and [3H]dexamethasone binding were studied in human renal adenocarcinoma and in uninvolved external cortex, in 22 patients undergoing nephrectomy for renal adenocarcinoma. Tissue incubations were performed with either [3H]aldosterone (5 x 10(-10) mol/l; 5 x 10(-9) mol/l in the presence of unlabelled glucocorticoids) or [3H]dexamethasone (5 x 10(-9) mol/l). 3. Cytosol [3H]aldosterone binding was six- to seven-fold lower (P less than 0.001) in neoplastic than normal tissue. [3H]Dexamethasone binding was about twofold higher in neoplastic than in normal tissue. This difference was not significant. 4. Nuclear uptake experiments showed that, both in cytosol fractions and nuclei, [3H]aldosterone binding was lower in adenocarcinoma than in normal cortex. 5. The very low binding of [3H]aldosterone suggests that mineralocorticoid receptors are absent in renal adenocarcinoma, an hypothesis in line with the proximal origin of these tumours.


Distribution of circulating immunoreactive components of parathyroid hormone in normal subjects and in patients with primary and secondary hyperparathyroidism: the role of the kidney and of the serum calcium concentration. 1. The distribution of intact parathyroid hormone-(1-84) [PTH-(1-84)] and of its COOH-terminal fragments was determined in human serum by column chromatography. In addition to PTH-(1-84) (peak I), COOH-terminal fragments having molecular weights of approximately 4000-7000 (peak II) and immunoreactive components co-eluting with human PTH-(1-12) (peak III) were observed. 2. Mean concentrations of intact PTH-(-84) and of its COOH-terminal fragments were significantly raised in chronic renal failure as compared with those of normal subjects. Mean amounts of peak II were higher in patients with chronic renal insufficiency than in nutritional vitamin D deficiency, in pseudohypoparathyroidism and in primary hyperparathyroidism, despite comparable amounts of PTH-(1-84). 3. In chronic renal failure as well as in a group of patients with vitamin D deficiency, pseudohypoparathyroidism and primary hyperparathyroidism and in controls, significant linear relations were found between the serum concentrations of calcium and log (peak II/peak I). Our findings suggest that the conversion of intact PTH-(1-84) into COOH-terminal fragments by the parathyroid glands (resulting in a raised secretion of fragments) and/or in peripheral organs may be directly related to the serum concentration of calcium. However, the degradation of the fragments may also be suppressed in a calcium-dependent manner.


Study of the oligosaccharide units from mucus glycoproteins of meconium from normal infants and from cases of cystic fibrosis with meconium ileus. 1. The mucus glycoproteins in meconium from normal infants and from infants having cystic fibrosis with meconium ileus have been studied. 2. Whereas normal meconium contained about 50% protein-bound carbohydrate, the meconium from cystic fibrosis contained only about 10%. 3. Glycopolypeptides were prepared from the mucus of glycoproteins. The oligosaccharide units from this material were released and fractionated. The fractions ranged widely in size and composition. 4. The fractions from cystic fibrosis specimens had a significantly higher content of fucose than those from normal specimens.


Lactoferrin turnover in man.

Lactoferrin turnover in man. 1. The turnover of 125I-labelled lactoferrin was measured in ten adults. Simultaneous organ radioactivity counting was performed. 2. Ferrokinetic studies were performed in three adults after the intravenous injection of [59Fe]lactoferrin. 3. Lactoferrin was rapidly eliminated from the plasma with a mean fractional catabolic rate of 5.7/day. Apolactoferrin (one subject) was eliminated at a slower rate (fractional catabolic rate 1.22/day). 4. Of the administered 125I label 99% was recovered in the urine, as free iodine, within the first 24 h. In the 59Fe studies no appreciable activity was found in the urine. 5. Organ radioactivity counting showed that lactoferrin was rapidly taken up by the liver and spleen. In the 125I studies the rapid excretion of free 125I suggested catabolism at these sites. In the 59Fe studies, the radioactivity persisted in the liver and spleen for several weeks and was slowly transferred to the bone marrow before appearing in circulating erythrocytes. 6. From the values of fractional catabolic rate, plasma lactoferrin, neutrophil lactoferrin and plasma volume, a 'derived neutrophil turnover' was calculated for each subject. The mean value was 8 x 10(8) neutrophils/day. This is about 1% the value obtained from the actual measurement of labelled cells. It is postulated that this 'derived value' represents only that portion of neutrophil turnover accounted for by intravascular senescence.


Studies on the renin-angiotensin-aldosterone system in elderly hypertensive patients with an angiotensin II antagonist. 1. To characterize the renin-angiotensin-aldosterone system in elderly hypertensive patients, an angiotensin II antagonist, [Sar1,Ile8]angiotensin II, was infused into individuals 60 years old and older with and without hypertension. 2. After infusion of [Sar1,Ile8]angiotensin II in all of the elderly patients and subjects an agonistic pressor response was observed that was greater than in middle-aged hypertensive patients. 3. Pre-infusion plasma renin activity and plasma aldosterone concentration in hypertensive and normotensive elderly groups were suppressed in comparison with those in middle-aged hypertensive subjects. The increased agonistic effects of [Sar1,Ile8]angiotensin II infusion on blood pressure in the elderly are presumably due to their hyporeninaemia. 4. The angiotensin-aldosterone system in elderly essential hypertensive patients is suppressed and is presumably not responsible for their elevated blood pressures.


Effect of propranolol on the ventilatory response to hypercapnia in man. 1. The effect of a single oral dose of propranolol on the ventilatory response to hypercapnia has been studied in five healthy subjects. 2. Propranolol produced a small but significant reduction in the slope of the ventilation/end-tidal PCO2 regression line. 3. Propranolol did not significantly change vital capacity or forced expiratory volume in 1 s.


Effect of general anaesthesia on renal haemodynamics in the rat. 1. The effect of sodium pentobarbital and Inactin anaesthesia on renal haemodynamics in the rat was evaluated with radioactive microspheres 15 micrometer in diameter. 2. Both anaesthetic agents caused substantial decrements in total renal blood flow (sodium pentobarbital, -34%; Inactin, -24%) compared with unanaesthetized animals. 3. Measurements of renal function obtained in rats anaesthetized with either of these anaesthetic agents should be interpreted with caution.


Mathematical analysis of the diagnostic relevance of clinical findings. 1. The process of diagnosis by elimination in terms of probabilities is analysed mathematically. 2. A piece of evidence which is only associated with a small number of differential diagnoses is of particular importance. 3. To deal with a differential diagnosis, a feature is required which occurs commonly in a postulated diagnosis but rarely in its rivals, irrespective of its frequency in those already discriminated against. 4. By using this analysis, inadequate evidence may be identified, new tests developed, and their effectiveness assessed in a logical and efficient way. 5. The approach is illustrated by considering the differential diagnosis of localized right lower quadrant abdominal pain.


Clonidine and sympathetic activity during sleep. 1. Blood pressure, heart rate, plasma noradrenaline and electroencephalographic variables were measured simultaneously during sleep in five healthy males before and after slow-release clonidine (300 micrograms orally) or placebo. 2. The blood pressure fall during sleep correlated significantly with the depth of sleep. 3. Clonidine significantly reduced systolic blood pressure during sleep and profoundly reduced total paradoxical sleep duration. 4. Plasma noradrenaline declined progressively during the sleep period and concentrations were significantly lower after clonidine compared with placebo.


Impairment of flow in routine gravity-fed intravenous infusions to surgical patients. 1. Gravity-fed intravenous infusions were studied in patients by using continuous flow recording and a new resistance-measuring method. 2. A common, though preventable, cause of reduced flow was the gradual deformation of the plastic tubing under the regulator clamp. 3. In many cases there was some constriction of the cannulated vein and in a few this constriction was severe enough to seriously impair the infusion. The severity of the constriction varied periodically. 4. There was no evidence of troble due to a generally raised pressure in the arm veins nor was clotting in the cannula a serious problem. 5. A simple test is described which may be found useful clinically for assessing the difficult infusion.


Release of dipeptide hydrolase activities from rat small intestine perfused in vitro and in vivo. 1. Hydrolase activities against three dipeptides were measured in mucosal cytoplasm in unperfused intestines and in mucosal cytoplasm, luminal effluents and serosal secretions after perfusion in vitro and in vivo for 1 h. Intestines in vitro were preparted both from anaesthetized rats and from freshly killed rats. 2. Only 0.6--1.9% of the initial cytoplasmic activity was recovered in the luminal effluent when intestines in vitro were prepared from anaesthetized rats. Recoveries in luminal effluents were similar (1.3--3.3%) during perfusion in vivo. 3. Losses of dipeptidases into the luminal effluent were four to eight times greater when intestines in vitro were prepared from freshly killed animals. 4. Similar losses of dipeptidases into the secretion on to the serosal surface were observed: they too were much greater when intestines were prepared from freshly killed animals. 5. Small losses of mucosal DNA during perfusion were also observed; however, losses of cytoplasmic peptidases were consistently slightly greater. 6. Enzyme loss therefore probably occurs both by sloughing of whole cells and by a more specific process which is greatly influenced by experimental procedure. Caution is necessary in the interpretation of peptide transport experiments in vitro, although the possibility that intraluminal hydrolysis is of physiological significance must not be excluded.


Plasma noradrenaline concentrations at different vascular sites during rest and isometric and dynamic exercise. 1. Blood samples were obtained simultaneously from femoral artery and vein, pulmonary artery and antecubital vein at rest and during isometric and dynamic exercise in six patients during cardiac catheterization and plasma noradrenaline was measured radioenzymatically. 2. Analysis of variance indicated that differences between vascular sites were not significant, whereas differences between patients (P less than 0.001) and differences between activities (P less than 0.001) were significant. 3. Mean systemic arterial concentrations tended to be lower than pulmonary arterial concentrations during each activity, although the difference was not significant. 4. We conclude that forearm venous plasma noradrenaline concentrations are representative of those from other sites.


CT scanning in the diagnosis of orbital disease. The application of CT scanning to the diagnosis of orbital disease is considered in the context of the general differential diagnosis of unilateral exophthalmos. The advantages of using a body scanner for orbital diagnosis are stressed, coupled with the high resolution facility available with the EMI CT 5005 machine. In addition to orbital space-occupying lesions and dysthyroid disease, secondary involvement of the orbit by paranasal sinus and middle fossa pathology is described, and the value of CT considered in respect to these lesions. The relationship of CT to other modalities of orbital investigation is discussed and their current place in orbital diagnosis evaluated.


Orbital syndromes--CT analysis of 100 cases. 100 cases of orbital syndromes, primary or secondary, are reviewed after CT analysis and compared with the results of plain and angiographic conventional examinations. First, the technique and normal results are described. Primary orbital syndromes (45 cases) originating in the orbit form three subgroups--tumors of the eyeball, tumors limited to the orbit and tumors of the orbit with bone lesions, with or without extra-orbital extension. Secondary orbital syndromes (55 cases) spreading to the orbit contain malignant tumors (36 cases) of which 25 are epitheliomas and benign tumors, the most frequent being meningiomas. For ocular and orbital tumors, CT allows the diagnosis of a mass and sometimes the pathological diagnosis (endocrine exophtalmos, inflammatory pseudotumors, varicose ophthalmic veins). For all other orbital tumors CT is important in determining the volume, relationships and extension, and does so better than carotid angiography or orbital phlebography. In secondary orbital syndromes, and particularly in facial malignant tumors and in meningiomas, CT is of great interest in the pretherapeutic evaluation of a tumor. It shows its volume and extension toward the face and orbital cavities, the pterygo-maxillary fossa (so important in determining the operability), the infra-temporal fossa and the endocranium, often without resorting to complementary investigational procedures which are much more aggressive. After histological diagnosis, CT allows the establishment of a therapeutic program.


Orbital trauma evaluation by computed tomography. Computed tomographic scans are useful for the evaluation of orbital trauma because they provide visualization of fractures, soft tissue damage, and embedded foreign bodies. Six cases are presented to demonstrate the usefulness of these studies in the evaluation of patients with a variety of injuries. By using both coronal and axial computed tomographic scans, orbital details can be studied with three-dimensional accuracy.


Advantages of a fourth generation CT scanner in the management of patients with orbital mass lesions. This presentation demonstrates the advantages of a fourth generation CT scanner in prognosis and localization of orbital space taking lesions, including its use as a localization guide for fine needle aspiration biopsy. It also provides an accurate means of follow-up of orbital lesions which have been either left in situ, partially excised, or excised. Six patients with difficult orbital problems of various types have been described to demonstrate how this high resolution CT scanner was utilized to improve the recognition and management of these problems.


Direct coronal computed tomography of the orbits employing a dedicated head scanner. A number of articles have been written on the topic of orbital computed tomography (CT), most of which have emphasized transaxial sections. Recently, especially since whole body scanners have become widely distributed, numerous articles have been written on coronal sections of the orbits. This article discusses the method of obtaining coronal CT sections of the orbits employing a dedicated head scanner and demonstrates the results that can be obtained employing this technique.


Computed tomography of the visual pathways. Many articles have discussed the anatomy and pathology of the orbits. Although this area is exquisitely visualized via computed tomography, the bulk of the visual apparatus lies behind the optic canal. A variety of pathologic conditions will manifest themselves by changes in the visual fields. This article demonstrates the normal anatomy of the visual pathways within the brain by CT scanning and correlates various changes that cause visual field defects.


Long-term survival of subacute cardiac rupture with tamponade in acute myocardial infarction, without surgical intervention (the role of pericardiocentesis). Four patients are described with cardiac tamponade caused by subacute cardiac rupture in acute myocardial infarction with long-term survival of two of them. Three patients were treated by pericardiocentesis. Clinical signs and symptoms and diagnostic problems are discussed stressing the need for a well-trained team of workers of the Coronary Care Units aware of the possibility of this event and ready to cope with its therapeutical demands--both surgical and conservative--by pericardiocentesis which is a small number of patients can be life-saving.


Changes in the pacemaker function of the sinus node after closure of atrial septal defect. Temporary and stable iatrogenic sick sinus syndrome. The alterations in the clinical symptoms and the pacemaker function of the sinus node following open heart operation were studied in 26 patients suffering from atrial septal defect. The most characteristic electrophysiological change was the prolongation of the sinus node recovery time as consequence of surgical repair of the atrial septal defect. The clinical symptoms of the sick sinus syndrome could be detected in all 26 cases after the open heart surgery. In 16 patients the symptoms of iatrogenic sick sinus syndrome (SSS) were only transitory, while in 10 patients a stable iatrogenic SSS developed after the operation. In the basis of these data the pathological mechanism of the iatrogenic SSS is discussed.


[A-wave in calibrated apex cardiogram in hypertension. Studies during rest and after exertion]. Apexcardiographic examinations were carried out of 86 hypertensive patients without clinically manifest heart failure. The amplitude of the wave a (or the per cent fraction of the wave a of the total amplitude) and the maximal rate of wave rise, df/dta were statistically significantly higher in the hypertensive patients than in normal subjects. The findings are interpreted as a manifestation of an increased left ventricular rigidity and of an increased atrial contractility in hypertensive patients. No correlation was found between the apexcardiographic indicators checked and the indicators of the cardiac pump function. In 46 hypertensive patients, in addition, a dynamic exercise test was performed (50 W for 6 min); 1 minute after cessation of exercise an apexcardiogram was recorded. An increase in the a/T quotient (greater than or equal to 5%) was found in 20; a decrease (=5%), in 10; and no change (less than +/- 5%), in 16 hypertensive patients. The postexercise apexcardiographic changes had no correlations with haemodynamic changes occurring during exercise.