The effect of recombinant human erythropoietin on hemostatic status in chronic uremic patients. The hemostatic effects of recombinant human erythropoietin (rHuEP) were investigated in 20 patients with end-stage renal disease (thirteen on hemodialysis, seven without dialysis) receiving this hormone. We studied their hemograms and coagulation profiles before and at 1 month after initiation of rHuEP therapy. One month after rHuEP administration, improvement in anemia (16/20, 80%) and shortening of bleeding time (17/19, 89.5%) were observed. Shortening or correction of bleeding time was achieved in three patients without any increase of the hemoglobin level. This means that factors other than the increased hematocrit level might contribute to shortening bleeding time in uremic patients receiving rHuEP treatment. The platelet count, prothrombin time, partial thromboplastin time, and fibrinogen level did not change over the course of rHuEP therapy. Thrombosis of vascular access was not observed, and heparin doses were not increased in this short-term period. A significant decrease was found in the plasminogen level, from 108.5% to 88.2% (p less than 0.05), in uremic patients on hemodialysis. The antithrombin III level also decreased, from 98.8% to 89.8% (p less than 0.05), and its level dropped to below normal ranges in six of thirteen patients (46%) on hemodialysis after treatment with rHuEP. No significant change was noted in the levels of antithrombin III, plasminogen, and alpha 2-antiplasmin in uremic patients not receiving dialysis. These results suggest that rHuEP administration induces increased extracorporeal dialyzer clotting and consumption coagulopathy, and that this extracorporeal consumption coagulopathy may play a role in the genesis of thrombotic complications.


Mullerian inhibiting substance binding and uptake. Mullerian inhibiting substance (MIS) is a 140,000 M(r) Sertoli cell derived glycoprotein with a critical regulatory role in the male fetus initiated presumably by ligand binding with receptor. To localize this binding species we performed time course incubations of cultured fetal rat lungs or control tissues with MIS, applied rabbit anti-MIS IgG, and fluorescein conjugated anti-rabbit IgG, and examined specimens with laser confocal microscopy. Punctate surface fluorescence followed by cytosolic and nuclear localization in lung consistent with specific adsorptive endocytosis was seen. Confocal imaging also detected MIS binding to the Mullerian duct in the urogenital ridge. Crosslinking of 125I-MIS with plasma membranes revealed a high molecular mass binder with signal displaceable by excess unlabeled ligand. These data support the hypothesis that a specific plasma membrane binding protein for MIS exists.


Effect of systemic calcium deficiency on the expression of transforming growth factor-beta in chick embryonic calvaria. The developmental process of intramembranous ossification involves bone formation directly from mesenchymal differentiation without a cartilage intermediate. We have previously observed that systemic calcium deficiency in the developing chick embryo, produced by long-term shell-less culture, results in the appearance of chondrocyte-like cells in the calvarium, a parietal bone which normally develops via intramembranous ossification. This investigation aims to analyze the mechanism underlying this calcium deficiency-related, aberrant appearance of cartilage phenotype in the chick embryonic calvarium. In view of the reported involvement of transforming growth factor beta (TGF-beta) in osteogenesis and chondrogenesis, we have examined and compared here the expression of TGF-beta in the chick embryonic calvaria of normal (in ovo development, NL), shell-less (SL), and calcium-supplemented SL (SL+Ca) embryos. TGF-beta expression was analyzed at the mRNA level by blot and in situ cDNA hybridization, and at the protein level by immunohistochemistry and immunoblotting. The results presented here indicate that: 1) TGF-beta is expressed in the chick embryonic calvarium by both periosteal cells and osteocytes, as revealed by in situ hybridization and immunohistochemistry; 2) TGF-beta expression is significantly increased in SL calvarium compared to NL calvarium, at both protein and mRNA levels; 3) the number of TGF-beta expressing cells increases in the SL calvarium, particularly along the central, subcambial core region of the bone; and 4) exogenous calcium repletion to the SL embryo affects the expression of TGF-beta such that the pattern approaches that in the NL embryo. Taken together, these results indicate that altered TGF-beta expression accompanies the aberrant appearance of cartilage phenotype caused by systemic calcium deficiency. We postulate that normal cellular differentiation along the osteogenic pathway during embryonic intramembranous ossification is crucially dependent on regulated TGF-beta expression.


Leg bud mesoderm retains morphogenetic potential to express limb-like characteristics ("limbness") in collagen gel culture. Recent in situ hybridization studies have correlated expression of potential regulatory genes with pattern formation in limb bud mesoderm (Tabin: Cell 66:199-217, 1991); however, the mechanism(s) controlling their expression in mesoderm and their relevance to the establishment of a limb morphogenetic pattern remain unknown. One likely candidate for regulating patterning events in limb mesoderm is the apical ectodermal ridge, as its removal in ovo results in a graded truncation of limb skeletal elements in the proximal-distal axis dependent upon the time of excision (Rowe and Fallon: J Embryol Exp Morph 68:1-7, 1982). In the present study, we investigate whether the hypothetical imprint of ridge ectoderm is retained in cultured mesoderm. Specifically, we sought to determine if a subpopulation of limb mesoderm that forms in collagen gel culture (Markwald et al: Anat Rec 226:91-107, 1990), retains any expression of "limbness" in the absence of limb ectoderm as characterized by the formation of a predictable number and distribution of limb-like chondrogenic elements in comparison to the temporal and spatial relationships of the in situ proximal, hindlimb skeletal structures. Accordingly, explants of undissociated mesoderm from stage 18-22 chicken leg buds were cultured without ectoderm on collagen gel lattices and the central subpopulation of mesoderm was examined morphologically. We show that this central subset of mesoderm will form chondrogenic cells which were not expressed uniformly throughout the subset, but rather distinct nodules or elements of cartilage were elaborated. Moreover, the number of elements expressed by the central subset increased with the age of the mesoderm at the time of explantation; spatially and temporally, the sequence of elements that formed always proceeded from the proximal, anterior margin of the subset to its distal, posterior border. The shapes of the initial elements (designated I and II) resembled the forms of in situ proximal skeletal structures (girdle and femur-like), whereas more distal elements (III-V) were often fused and without structural similarity to in situ skeletal structures. When cultures were established from the posterior mesoderm of stage 19/20 or 21 mesoblasts, the frequency of element I formation was reduced approximately one-half, whereas formation of more distal elements was unaffected. Conversely, element formation from the central subset established from isolated anterior mesoderm was virtually identical to intact mesoblasts, indicating a capacity to regulate for the loss of mesoderm as occurs in situ (Hampé: Archs Anat Microsc Morph Exp 48:345-378, 1959).(ABSTRACT TRUNCATED AT 400 WORDS)


Developmental expression of two antigens associated with mouse intestinal crypts. Two monoclonal antibodies were prepared against the duodenal mucosa of four-day-old mice (MIM-1/39 and MIM 1/130). The expression of the antigens was associated with the crypts of the small and large intestine in the fetus and adult. MIM-1/39 was present in epithelial cells of the intervillous areas in the small intestine at 17 and 18 days of gestation; afterwards its expression was detected only in crypt cells from birth to adulthood. Transition from the mouth of the crypts to intestinal villi was abrupt. Expression of MIM-1/39 was first detected at time of birth in the colon: In the adult, only crypt cells expressed the antigen and goblet cells were negative. Antigen MIM-1/130 was detected from 16 to 18 days of gestation in the small intestine, in the mesenchymal matrix lying under the intervillous epithelium. After birth, it was present in the pericryptal mesenchymal matrix. This antigen was also expressed at birth in the colon and remained in the pericryptal matrix in the adult. In vivo, multiple injections of an organic extract of rat amniotic fluid to mothers, starting at 14 days of gestation, induced a profound modulation in the pattern of expression of both antigens at 17 days of gestation: The pattern of expression was comparable to that observed at least 5 days after birth in untreated animals. The expression of both antigens before crypt appearance may reflect some molecular differentiation in preparation for the formation of crypts, while their association with differentiated crypts may indicate that they have a role in the maintenance of crypt functional and/or morphological integrity.(ABSTRACT TRUNCATED AT 250 WORDS)


Expression of alpha-cardiac and alpha-skeletal actin mRNAs in relation to innervation in regenerating and non-regenerating rat skeletal muscles. The expression of alpha-cardiac and alpha-skeletal actin mRNA in regenerating muscle was examined. Changes in mRNA levels were analyzed in autografted extensor digitorum longus (EDL) muscles in rats using alpha-isoform specific synthetic oligonucleotides and beta-actin cDNA as probes. After autografting, the expression of alpha-cardiac actin mRNA was induced; concomitantly that of alpha-skeletal actin mRNA was reduced. The pattern of alpha-actin mRNA expression appeared to be similar to that seen in embryonic skeletal muscle. In order to evaluate the effects of innervation on alpha-actin mRNA expression in regenerating muscle, nerveless, standard, and nerve-intact autografted muscles were examined. More complete innervation facilitated the recovery of alpha-skeletal actin mRNA to control levels, but had little effect on the amount of alpha-cardiac actin mRNA. We found that regenerating muscle shows that embryonic pattern of alpha-actin mRNAs in the early stage and concluded that the recovery of alpha-skeletal actin mRNA expression to the adult pattern is influenced by innervation, while alpha-cardiac actin mRNA expression is nerve independent.


Sense and antisense TGF beta 3 mRNA levels correlate with cardiac valve induction. The formation of the valves in the heart is a spatially and temporally controlled process. A tissue interaction between the endothelium and its adjacent myocardium initiates the transformation of the endothelium into the mesenchymal precursors of the heart valve. One or more of the molecules implicated as critical for valve formation are members of the transforming growth factor beta family of molecules. Presented here is a spatial and temporal analysis of TGF beta 2 and TGF beta 3 in the chick heart during valve formation. We show that TGF beta 3 mRNA is concentrated in AV canal tissue where valve formation will occur, consistent with previous observations that TGF beta 3 production is critical during valve formation. Additionally, an RNA complementary to TGF beta 3 encoding mRNA is present in the heart. The temporally controlled appearance of RNA complementary to TGF beta 3 suggests that this molecule may play a role in the regulation of TGF beta 3 production in the heart.


Differences in the histogenesis of EDL and diaphragm in rat. We have examined the histogenesis of the diaphragm and extensor digitorum muscle in rat embryos, with the aim of defining differences in developmental patterns that can be related to the functional requirements of these muscles during and after development. Patterns of interactions between myotubes and other cells, and frequency of gap junctions are quite different in the two muscles. In diaphragm, primary myotubes (at day 16 in utero) are closely associated with each other, forming parallel sheets or palisades and communicating by gap junctions. Secondary myotubes have formed by day 18, but are immature, and the frequency of gap junctions is lower. The arrangement in palisades is maintained even after fibers are separated from each other by their individual basal lamina. In EDL primary fibers at day 16 have fewer gap junctions, and the peak in communication occurs after the appearance of secondary myotubes (day 18 and 21). Secondary myotubes are more mature than in diaphragm at day 18.


Changes in dorsoventral but not rostrocaudal regionalization of the chick neural tube in the absence of cranial notochord, as revealed by expression of engrailed-2. Notochord has been implicated in previous studies in both the dorsoventral and rostrocaudal patterning of the developing neural tube. This possibility has been further explored by analyzing the expression of Engrailed-2 in chick embryos developing with cranial notochord defects. Control embryos containing intact notochords expressed Engrailed-2 protein within the neural tube and in a subset of the neural crest and overlying surface ectoderm at the future mesencephalon and cranial metencephalon levels. Within the neural tube, expression was confined to cell nuclei in the roof plate and lateral walls; floor plate nuclei directly overlying the notochord typically failed to show expression. After surgical removal of Hensen's node, the source of notochord precursor cells, embryos were cultured through neurulation and assayed for expression of Engrailed-2 protein. All embryos that partially or completely lacked cranial notochord expressed Engrailed-2 in a pattern similar to that of control embryos containing intact notochords, except that when notochord and floor plate were absent, Engrailed-2 was also expressed in the most ventral part of the neural tube. These results indicate that 1) Engrailed-2 expression is suppressed in the most ventral neural tube owing to induction of the floor plate by the notochord, and 2) that the presence of an underlying notochord is not required for correct rostrocaudal expression, suggesting that multiple pathways act in the patterning of the rudiment of the central nervous system.


Viruses and the blood.

Viruses and the blood. Haematological syndromes attributed to viruses demonstrate geographical variations in incidence and great dependence on host factors. Severe haematological disease is the exception rather than the rule in dengue virus infection, and probably depends at least in part on the host immune response to the virus. The increased incidence of hepatitis-associated aplasia in east Asia may reflect distribution of an infectious agent, an environmental toxin, or genetic predisposition, but probably represents some combination of these factors. Agents with apparently universal distribution, such as parvovirus B19 and Epstein-Barr virus, are associated with bone marrow failure only in a very narrow range of hosts. These examples teach us that viral causes cannot automatically be excluded from the differential diagnosis of syndromes whose occurrence is rare or apparently sporadic. Further investigation of these syndromes should include more detailed characterization of host factors, particularly immunological characteristics, and possible infectious and toxic cofactors which are associated with morbidity.


Anaemia of Plasmodium falciparum malaria. The pathophysiology of the anaemia of falciparum malaria is both complex and multifactorial, and results in a condition which is a major cause of mortality and morbidity in patients, especially children and pregnant women, living in malarial endemic areas. The importance of anaemia as a cause of death in malaria may well be underestimated because of difficulty in diagnosis, especially where parasitaemia may be low and the clinical picture may be confused with other causes of anaemia. Two clinical presentations predominate: severe acute malaria in which anaemia supervenes, and severe anaemia in patients in whom there have been repeated attacks of malaria. The major mechanisms are those of red cell destruction and decreased red cell production. Potential causes of haemolysis include loss of infected cells by rupture or phagocytosis, removal of uninfected cells due to antibody sensitization or other physicochemical membrane changes, and increased reticuloendothelial activity, particularly in organs such as the spleen. Decreased production results from marrow hypoplasia seen in acute infections, and dyserythropoiesis, a morphological appearance, which in functional terms results in ineffective erythropoiesis. The role of parvovirus B19 as a possible cause of bone marrow aplasia in a few cases is postulated. Finally, there is now evidence which points to genetic factors, HLA associated, which may protect against the development of malarial anaemia and which has become common in areas endemic for malaria.


Genetic epidemiology of the beta s gene. The beta s gene arose at least four times in Africa, with three of these mutations expanding through diverse ethnic groups, but limited to definite geographical areas: Atlantic west Africa for the Senegal haplotype linked beta s; central west Africa for the Benin haplotype; and equatorial, eastern and southern Africa for the Bantu haplotype. The fourth mutation (linked to the Cameroon haplotype) is restricted to a single ethnic group, the Eton of central Cameroon. The Benin haplotype linked beta s gene was spread by gene flow to the Mediterranean (north, south and east) and to the western portions of Saudi Arabia. An independent mutation linked to a fifth haplotype, Arab-India, is found among the tribals of India (independent from their geographical origin) and in the eastern oases of Saudi Arabia. It is also suspected of being associated with the beta s gene found in Afghanistan, Iran, Transcaucasia and central Asia. The selective force involved in the expansion of the gene was most likely P. falciparum malaria, and the time of the gene frequency increase was likely to have been during the expansion of agriculture about 4000 or more years ago in India and about 3000 years ago in Africa. The partial protection against severe and life-threatening malaria is through the limitation of P. falciparum parasitaemia. This is a complex process which involves at least two mechanisms: early intraerythrocyte parasite forms are in a suicidal position through increasing the tendency of HbAS cell to sickle and then be destroyed by the spleen; intraerythrocyte growth is inhibited during deep vascular schizogony. Although there is evidence that P. falciparum (and P. malariae) parasitaemias are limited in HbSS red cells, malaria is a major trigger to haemolytic and infarctive crises in sickle-cell disease, and a common cause of morbidity and mortality.


Glucose-6-phosphate dehydrogenase deficiency. Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzymopathy. Because its gene locus is on the X-chromosome it is more common in males than females in all populations. Prevalence rates vary from 62% among Kurdish Jews to the very low rates (0.1% or less in Japan, for example), which are compatible with sporadic cases arising from spontaneous mutations. However, there is at least one population in which G6PD deficiency has not been found, namely the indigenous (Amerindian) population of America. Approximately 400 variants have been described. Despite the clinical burden imposed by this enzymopathy, polymorphic frequencies have been reached in many populations. There is abundant epidemiological evidence that this has happened because of a biological advantage conferred on heterozygotes in falciparum malaria endemic areas. This advantage may apply to quartan malaria as well. Clinical severity varies, from the rare chronic nonspherocytic haemolytic anaemia to progressively milder forms like the Mediterranean and A- types. The other clinical syndromes, i.e. neonatal jaundice and haemolysis caused by infections, foods, drugs and chemicals, are not always predictable. This is because only a fraction of such enzymopathic persons develop these syndromes after exposure to the relevant stimulus. Modern techniques of molecular biology may elucidate why this is so. There is some emerging evidence that the genetic burden or survival value associated with G6PD deficiency may be relevant not only in tropical and infectious diseases, but also in their chemotherapy (e.g. malaria) as well as in the control of a long-recognized environmental pollutant such as lead.


Platelets and platelet disorders in Africa. Blood platelets, which are known to play important roles in normal vertebrate biology, are influenced by a variety of factors, the majority of which are acquired. In this chapter, attention is drawn to the need to re-examine one of the old accepted values in haematology, the normal platelet count. In addition, effects of some environmental factors, such as malarial parasitaemia, on both platelet count and platelet function are fully discussed. Similarly, the effects of HIV on platelets, as well as on other haematological parameters, in the African environment are described briefly. The urgent need is stressed for careful evaluation of the impact of HIV on the total disease burden in the population. In addition, the effect of environmental factors on the manifestations of some clinical features, e.g. splenomegaly and anaemia, in acute idiopathic thrombocytopenic purpura are highlighted. It is suggested that in such environments, the presence of these features do not constitute contraindications to making the diagnosis when other criteria are met.


Onyalai.

Onyalai. Onyalai is an acquired form of immune thrombocytopenia which differs clinically, epidemiologically and immunologically from idiopathic thrombocytopenic purpura (Table 4). The clinical hallmark is haemorrhagic bullae on the mucosa of the oronasopharynx. Haemorrhage from ruptured bullae, epistaxis or gastrointestinal bleeding is severe and may cause shock and death. The disease is limited to some black populations of central southern Africa, with a recorded incidence of one per 660 inhabitants per year in the Kavango territory of Namibia. The majority of patients demonstrate both IgG and IgM serum platelet antibodies and serum platelet glycoprotein IIb/IIIa autoantibodies. Chronic thrombocytopenia often ensues and recurrent episodes of clinical bleeding are common. Treatment directed at the prevention of haemorrhagic shock reduced the mortality rate in the acute phase from 9.8 to 2.8%. Standard dose prednisolone does not increase the platelet count. Vincristine sulphate may benefit some patients and splenectomy is indicated in patients with severe uncontrollable haemorrhage. High dose intravenous gammaglobulin may be followed by a rise in the platelet count and cessation of haemorrhage. The aetiology is unknown. The possible aetiological role of mycotoxins from contaminated millet, sorghum or maize requires further investigation.


Chromatin loops and transcriptional regulation. The existence of a supermolecular structure in the eukaryotic nucleus, involving loops of chromatin attached at irregularly spaced points to the nuclear matrix, is now well established. Quite a bit is known concerning the DNA sequences involved in the attachment. However, the function of this highly organized structure remains largely unknown. In this review, we attempt to provide an overview of present knowledge of nuclear loop structure, and to critically summarize recent studies that provide new insights as to function. We derive the conclusion that the loop structure is very likely one in which DNA topology can be locally regulated by topoisomerase activity, and is most probably modified during development.


c-myb and growth control.

c-myb and growth control. The available evidence indicates that c-myb plays an important role in the proliferation of hematopoietic cells and in those nonhematopoietic cell types in which c-myb is expressed. A critical aspect in the regulation of c-myb expression rests in the positive autoregulatory mechanism, which is dependent on the interaction of myb protein with the 5' flanking region of the human c-myb gene. The positive autoregulation of c-myb, in conjunction with tissue-specific mechanisms that most likely involve efficient transcription beyond the site of "transcriptional pause" in the c-myb first intron, might allow the generation of c-myb transcripts at levels sufficiently high for optimal biological activity (e.g., at the G1/S transition of the cell cycle). Other transactivating factors, such as the Jun family members, also appear to be involved in regulating c-myb expression. Such factors might act to increase basal levels of c-myb expression to allow activation of the autoregulatory mechanism, or might cooperate with myb in transcriptional regulation of c-myb expression. The function of c-myb is ultimately dependent on the genes that are regulated by the myb product. Preliminary evidence suggests that DNA polymerase-alpha and cdc2, two genes that are critical for DNA synthesis, contain myb binding sites in their promoter region that appear to be required for myb transactivation of their expression. The paradox of the generality of the mechanisms by which c-myb affects cell proliferation and the apparent tissue-specific expression of this gene might be resolved by the growing evidence that the tissue distribution of c-myb is more general than previously appreciated, and that many cell types with no detectable c-myb expression contain a functional equivalent of this gene. For example, B-myb a gene that is homologous to c-myb in the DNA binding and transactivating domains and appears to be ubiquitously expressed, is also required for cell proliferation and, like c-myb, appears to regulate the expression of cdc2, a gene required for cell cycle progression. Together, these findings indicate a general role of members of the myb family in regulation of cell proliferation.


Regulation of terminal deoxynucleotidyl transferase gene expression in mice and men. A nontemplate directed DNA polymerase, terminal deoxynucleotidyl transferase (terminal transferase) is expressed in a tissue-specific and development stage-specific manner. Its enzymatic properties and tissue localization have implicated the protein in development of normal immune function. Significant progress has been made in understanding the enzymology and important domains of this protein. More recently, studies have focused on regulation of the gene that codes for the protein in mice and humans. The murine gene has yielded to these studies more readily than the human gene. A murine basal promoter element has been identified along with several trans-acting protein factors that may regulate gene expression. In the human gene there is no evidence for a basal promoter element. Rather, the promoter exhibits tissue specific properties. The present article reviews recent developments in this field.


Cell cycle regulation and the p53 tumor suppressor protein. Somatic mutations of the p53 gene have been implicated as causal events in the formation of a large number of common human tumors. Several lines of evidence suggest that the nuclear phosphoprotein encoded for by the wild-type gene (wt-p53) plays a role in regulating cell proliferation. Wt-p53 protein encodes a potent negative growth regulatory function that is lacking in mutant forms of the protein found in human tumors. In this review, the relationship between the expression of wt-p53 protein and cell proliferation is examined with emphasis on recent studies that provide clues as to the possible role that p53 plays in cell cycle regulation. A model for the action of p53 in regulating cell proliferation is proposed in which wt-p53 acts as a "checkpoint" protein to control the transit of cells through the restriction point in late G1-phase. After cells pass this "checkpoint" they become committed to enter S-phase and initiate DNA replication. This checkpoint function may be defective in cells that lack p53, express mutant p53, or in which the antiproliferative form of the protein is functionally inactive. Under these conditions stringent control of the initiation of DNA replication may no longer be possible, providing an environment conducive to the emergence oncogenic clones.


Reference intervals and developmental changes in urinary prostanoid excretion in healthy newborns, infants and children. Urinary excretion of prostaglandins E2, F2 alpha, E-M (7 alpha-hydroxy-5, 11-diketotetranor-prosta-1, 16-dioic acid), 6-keto F1 alpha, 2,3-dinor-6-keto-F1 alpha, thromboxane B2, 2,3-dinor-thromboxane B2 and 11-dehydrothromboxane B2 was determined by gas chromatography-mass spectrometry in 83 healthy subjects aged one day to 37 years. The excretion rates of all prostanoids increased with advancing age. After correction for 1.73 m2 body surface area, only urinary excretion rates of prostaglandins E-M and 6-keto-prostaglandin F1 alpha depended on age. Reference intervals were calculated as the 10th and 90th percentiles for prostaglandins E2 (4-27 ng/h/1.73 m2), F2 alpha (23-87 ng/h/1.73 m2), 2,3-dinor-6-keto-F1 alpha (4-19 ng/h/1.73 m2), thromboxane B2 (1-21 ng/h/1.73 m2), 2,3-dinor-thromboxane B2 (8-36 ng/h/1.73 m2) and 11-dehydro-thromboxane B2 (15-87 ng/h/1.73 m2) in all subjects, and for prostaglandins E-M and 6-keto-prostaglandin F1 alpha in subjects aged 30 days or less (110-1140 ng/h/1.73 m2 and 7-23 ng/h/1.73 m2) and older than 30 days (62-482 ng/h/1.73 m2 and 2-12 ng/h/1.73 m2). High urinary excretion of prostaglandins E-M and 6-keto-F1 alpha during the newborn period and some distinct changes in urinary excretion of prostaglandin E2 and thromboxane B2 with advancing age suggest that these prostanoids might play a specific role during child development.


Psychological development in children born with very low birth weight after severe intrauterine growth retardation: a 10-year follow-up study. Children born small for gestational age (SGA) and children having very low birth weight, less than 1500 g, are claimed to be at risk of developmental problems, even when obvious pathology and disability are absent. In this study, sensorimotor and cognitive development of 14 medically healthy, very-low-birth-weight and small-for-gestational-age children were investigated. The children were born at the Karolinska Hospital between 1979 and 1981. At the time of the assessment, the children were aged 8.7-11.2 years. The assessment instruments included the Wechsler Intelligence Scale for Children, a modified version of the Bruininks-Oseretsky Test of Motor Proficiency, as well as selected subtests from the Halstead-Reitan Neuropsychological Battery and from the Southern California Tests of Sensory Integration. Information was also obtained from obstetric, neonatal and pediatric records, which included early developmental assessments. As a control group, 14 children were recruited and matched for age, sex and socio-economic background. The very-low-birth-weight-small-for-gestational-age group scored significantly lower on measures of visuospatial ability, non-verbal reasoning, strategy formation and gross-motor coordination. The group differences were largely attributable to the subnormal performance of eight of the very-low-birth-weight-small-for-gestational-age children. These children, who also tended to be born earliest (less than 33 weeks), had a high incidence of behavioral and educational problems. These findings are consistent with the view that the very preterm infant develops a different neurobehavioral organization than a full-term infant. Developmental deficits may become increasingly evident in the early school years.


Fluid restriction for preventing bronchopulmonary dysplasia? Reduced fluid intake during the first weeks of life improves the outcome of low-birth-weight infants. One hundred consecutive low-birth-weight infants (less than 1751 g) were randomized equally into "dry" and "control" groups. Fluid administration in the dry group followed the schema 50, 60, 70, 80, 90, 100 and 120 ml/kg/day during the first week and 150 ml/kg/day until aged four weeks and that of the control group 80, 100, 120 and 150 ml/kg/day during the first week and 200 ml/kg afterwards. The diagnosis of bronchopulmonary dysplasia was based on typical roentgenographic findings, clinical symptoms and a supplemental oxygen requirement at age one month. Mean weight loss was greater in the dry group than in the control group (8.8 vs 6.2%), but there was no difference in weight between the groups at the age of four weeks. Twelve of the patients died, one in the dry group and 11 in the control group (five died during the first 24 h and the remaining six between three days and three months). Twenty-seven subjects in the dry group and 15 in the control group were alive and had no signs of bronchopulmonary dysplasia at age 28 days (p less than 0.05), the numbers at term being 28 and 14, respectively (p less than 0.01). These results suggest that fluid restrictions can reduce the mortality and morbidity of low-birth-weight infants. Infants of this kind, with respiratory distress, show poor tolerance to fluids, especially during the first weeks of life.


Treatment with hypertonic dextrose and insulin in severe hyperkalaemia of immature infants. Over a three-year period, 12 infants received dextrose/insulin infusions for severe hyperkalaemia from a mean age of 24 h. The infants were born after 24 to 26 weeks of gestation and weighed 730 +/- 172 g (mean +/- SD) at birth. Serum potassium concentration ranged from 7.4 to 8.4 mmol/l (7.7 +/- 0.4 mmol/l; mean +/- SD). Four had cardiac arrhythmias. All infants showed an initial response, serum potassium concentration decreased below 6.5 mmol/l in 5 +/- 2 h. In two infants, rebound hyperkalaemia occurred and was resistant to treatment; both infants died, one during an exchange transfusion. In the other 10 infants, infusions were ceased at a mean postnatal age of 53 h. Hyperglycaemia was the major problem during infusion and was resistant to increases in insulin concentrations. Normoglycaemia was achieved in 10 infants. The hypertonic solution consisted of a dextrose/insulin ratio of 2.2 +/- 0.6 g/IU, which delivered glucose at a rate of 0.46 +/- 0.15 g/kg/h, in addition to the pre-existing stable maintenance glucose intake.


Pneumonia and pneumonitis in childhood malignancy. We conducted a survey over a 21-year period of the incidence and risk of occurrence of episodes of pneumonia and pneumonitis in children treated for solid tumours and leukaemia. One hundred episodes occurred amongst 219 patients, seven of which were associated with death. Focal opacification on the chest radiograph was more common than diffuse opacification. Patients with leukaemia had a significantly higher rate of occurrence of pneumonia and pneumonitis during the periods of induction and maintenance compared with the off-treatment period, and during the relapse period compared with the period of maintenance. Patients with solid tumours had a significantly higher rate of occurrence during treatment compared to the off-treatment period. The rate of occurrence on treatment was the same in patients with solid tumours and acute leukaemia. Children with malignancy have a high incidence of pneumonia and pneumonitis and death is rare if the patient does not have terminal malignant disease.


Modification of some markers of inflammation during treatment for acute respiratory exacerbation in cystic fibrosis. An objective approach for monitoring the treatment of acute pulmonary exacerbation in cystic fibrosis was evaluated. Eleven biochemical markers of inflammation (erythrocyte sedimentation rate, neutrophil count, C-reactive protein, alpha-1 antitrypsin, haptoglobin, ceruloplasmin, fibronectin, alpha-1 glycoprotein, alpha-2 macroglobulin, C3, granulocyte elastase and anti-Pseudomonas IgG) were measured in blood serum and plasma from 46 cystic fibrosis patients with chronic Pseudomonas aeruginosa colonization before and after treatment. The overall outcome in each patient was evaluated by means of a pondered sum of clinical, chest X-ray and lung function scores. Biochemical markers were related to the overall clinical improvement: haptoglobin, ceruloplasmin, fibronectin and alpha-1 glycoprotein showed a good sensitivity (64-70%), specificity (60-70%) and positive predictive value (86-89%). Granulocyte elastase showed a similar sensitivity (67%) and positive predictive value (85%) but a lower specificity (33%). The negative predictive value was generally poor (32-39%). Our data suggest that the combined measurement of some markers of inflammation and of conventional clinical parameters, may help in evaluating the efficacy of anti-infective treatment in cystic fibrosis.


Tracheostomy in pediatric patients.

Tracheostomy in pediatric patients. From 1978 to 1987, tracheostomy was performed on 33 children, 13 boys and 20 girls, with a male to female ratio of 0.65:1. The mean age at the time of tracheostomy was 726 days, 76% of the children being under the age of two years. The incidence of pediatric tracheostomy per hospital admissions was 0.05%. Subglottic stenosis (13 children) and respiratory distress syndrome with prolonged endotracheal ventilation (11 children) were the most common indications for tracheostomy. The mean duration of prolonged endotracheal intubation before tracheostomy was 64 days, and that of tracheostomy treatment 117 days. During the tracheostomy period, five children died, but only one death was related to tracheostomy. The total rate of complications was 30%. We emphasize the importance of strict indications for pediatric tracheostomy.


Late sequelae of gonadal, mediastinal and oral teratomas in childhood. Thirty-two patients with gonadal or non-sacrococcygeal extragonadal teratomas who underwent operation in childhood were evaluated for different late sequelae. Follow-up times ranged from 5.0 to 40 years. Physical abnormalities resulting from surgery were seen in eight patients who had had oral or mediastinal primary teratomas. Radiographs revealed multiple spina bifida occulta lesions in three patients. The males, but not the females, showed signs of gonadal dysfunction. Impaired semen quality or abnormal serum concentrations of testosterone or gonadotrophins were found in five of 10 adult men who had had testicular or mediastinal teratomas. Thus, some late sequelae may be predicted from the site of the primary teratoma, but gonadal dysfunction in males and vertebral anomalies may occur irrespective of where the teratoma is situated.


Metabolic control and diet in Finnish diabetic adolescents. The effects of dietary, clinical and demographic factors on metabolic control in 105 diabetic adolescents were studied. All patients had diabetes for longer than two years and a daily insulin dose greater than 0.5 IU/kg body weight. Low body mass index, high social class, high number of daily eating occasions, high day-to-day variation in energy intake, high number of urine tests and a long interval between insulin injection and eating were associated with good metabolic control. Many of these determinants reflect also the general compliance with the diabetic regimen. The results stress the importance of good coordination between insulin injections and eating habits.


A single pediatric center experience with 1025 children with hypertension. Between January 1982 and December 1989 1025 patients aged between one month and 18 years with increased blood pressure were referred for evaluation. Borderline hypertension was found in 389 children; 636 had sustained significant hypertension. In 351 patients, hypertension was secondary to a known disease. Renal parenchymal diseases were present in 68% of patients while renovascular and endocrine disorders were found in 10% and 11%, respectively. Of the 258 children aged less than 15 years, all but six children had known causes of hypertension, while 75% of adolescents had essential hypertension. In the 389 children with borderline hypertension, 65% developed fixed hypertension over a period of 2-3 years.


Doppler echocardiographic and morphologic evaluation of patients following operative repair of aortic coarctation. Sixty-six patients operated on previously for coarctation of the aorta were included in the study. There were 50 children and 16 adults. Invasive and non-invasive gradients were recorded. Anatomical obstruction was evaluated by angiography in 54 patients and the numbers and size of collaterals were quantified. The closest correlation was found between Doppler-estimated gradients and invasive peak-instantaneous gradients (r = 0.87). The relation between anatomical obstruction and invasive peak-to-peak gradients, Doppler gradients and diastolic flow ere discouraging (r = 0.64, 0.54 and 0.35). Liberal indications for angiography or magnetic resonance tomography are recommended.


Medical vocational guidance for adolescents--is it effective? The outcome of school medical vocational guidance was studied in all 1595 students enrolled in 1987 in grade 9 in the primary schools of Linköping. In grade 9, the school health service had listed 249 students with a chronic disorder. These were sent a questionnaire which was answered by 235 students. Only 5/235 stated that vocational information from physicians and school nurses had been important for their choice of career. No more than 22/235 had chosen a medically ill-suited secondary school study course. This figure is close to the number expected, assuming that all choices of study courses are at random. At follow-up in May 1989, no one said that they had changed study course or work due to their medical disorder. Thus, school medical vocational guidance seems to have little effect on the vocational choices of adolescents with chronic disorders.


Health examination of preschool children in relation to school adjustment in grade I. The results of a collaborative study on health examination in preschool children, using data from child health care, preschool establishments and a teacher evaluation carried out six months after the start of school were compared. The analysis showed that many children with difficulties in motor function, behaviour and/or learning in grade 1 had been identified at the health examination. The results emphasize the value of collaboration between preschool establishments, child health care centres and schools for identification of children at risk of developing early adjustment problems.


Thyroid dyshormonogenesis: severe hypothyroidism after normal neonatal thyroid stimulating hormone screening. We report four children who presented no evidence of primary hypothyroidism in the neonatal period, either clinically (normal growth velocity) or biochemically (normal plasma levels of thyroid stimulating hormone and/or thyroid hormone). However, in early childhood, these children developed severe hypothyroidism due to dyshormonogenesis. We conclude that apparently normal thyroid function in the neonatal period does not preclude the development of severe hypothyroidism due to thyroid dyshormonogenesis later in childhood.


The value of peritoneal dialysis in the treatment of severe ethanol intoxication in childhood revised. We treated a girl aged 3.5 years (15 kg) with ethanol intoxication, using peritoneal dialysis. The blood ethanol concentration was 6.4 g/l (640 mg/dl; 138.9 mmol/l). It was calculated that the child drank a total amount of 67.2 g of ethanol (4.5 g/kg). The spontaneous ethanol elimination rate before peritoneal dialysis was 0.27 g/l (5.86 mmol/l) per hour; during peritoneal dialysis we found an ethanol elimination rate of 0.32 g/l (6.94 mmol/l) per hour, which was lower than expected. In childhood the ethanol elimination rate with peritoneal dialysis is only slightly faster in comparison to the high spontaneous elimination rate. We conclude that treatment of severe ethanol intoxication should include mainly the maintenance of the vital functions and the meticulous control of blood sugar levels and acid-base disturbances, especially in children. Indications for dialysis are complications caused by ethanol and resistant to supportive therapy, such as seizures, metabolic disturbances, persistent hypoglycemia and the possibility of combined intoxication with other dialysable drugs.


[Therapeutic embolization of pulmonary arteriovenous malformations]. It is reported on the practicability and the results of therapeutic embolizations in 7 patients with pulmonary av-malformations. The embolizations were carried out by detachable balloons. In all cases there were immediate and well-defined occlusions. In direct fistulas the embolization represents a curative procedure. However, in cases of angiomatous malformations with slow flow character, only a reduction of shunt or decreasing of size is obtained.


[Internal carotid artery aneurysm in the petrous canal]. Internal carotid aneurysm in the petrous canal are rarely observed. According to our knowledge there are only 24 reported cases. We present the case of a 47-year old woman. The clinical, radiological and histological findings are discussed.


[Magnetic resonance tomography: value and limits in gastroenterologic diagnosis]. At present, the value and limitations of magnetic resonance imaging (MRI) as a diagnostic modality in gastroenterology cannot be conclusively determined. MRI has a high sensitivity in the detection of focal liver lesions. The signal intensity of focal liver lesions on plain and contrast-enhanced T1-weighted images and on plain T2-weighted scans narrows the differential diagnosis in many cases. Imaging of the gallbladder and pancreas presently is no indication for MRI. The high sensitivity of MRI in the detection of fistulae and abscesses in Crohn's disease and the differentiation from fibrosis represents an advantage over other modalities in imaging of inflammatory bowel disease. MRI is superior to CT in differentiating recurrent rectal cancer from fibrosis. Further development of MRI and the use of contrast media for both oral and intravenous administration may offer new perspectives of MRI in the diagnosis of diseases in gastroenterology.


[Controlled percutaneous puncture of gastrointestinal and hepatopancreatic lesions. An overview]. The cross sectional imaging of ultrasonography and computed tomography provides information for the puncture of focal lesions and fluid collections and additional drainage if necessary. A survey is given of the predispositions, technical aspects, results and complications of both methods.


[Endosonography in malignant extrahepatic obstructive jaundice]. 34 patients with obstructive jaundice were investigated consecutively by transcutaneous ultrasonography, endoscopic sonography and endoscopic retrograde cholangio-pancreatography (ERCP). In 20 cases obstruction was due to a tumor, whose location and extent could be visualized by endosonography and ERCP in all cases. By transcutaneous ultrasonography this was possible in only 14 (70%) and 17 (85%) cases respectively. In seven of eight cases the tumor size, in four of five cases metastatic involvement of lymph nodes and in all cases infiltration into neighboring organs or vessels were correctly predicted by endosonography. Both other methods failed in this regard. We therefore conclude that the diagnostic information obtained by endosonography in patients with obstructive jaundice due to malignancy is superior to conventional sonography and ERCP.


[Splenic size and duplex sonography determination of blood flow in the vena lienalis and vena portae in liver cirrhosis]. Splenomegaly is a common finding in patients with portal hypertension. In the present study the relation between spleen size and blood flow in the splenic and portal vein was evaluated in 33 patients with alcoholic liver cirrhosis and portal hypertension using pulsed Doppler sonography (Ultramark 9, ATL, Solingen, FRG). There was a significant positive correlation between hilar spleen diameter (HD) and splenic vein diameter (r = .73, p less than .001) as expected as the consequence of portal hypertension. However, a positive correlation between HD and splenic vein flow (SBF) was found (r = .67, p less than .001). Furthermore, there was no negative correlation between HD and flow velocity in the splenic vein (r = .01, n.s.). Portal blood flow (830 +/- 360 ml/min) was fairly constant in spite of considerable variations in SBF (range: 120 to 1200 ml/min). The data of the present study indicate that splenomegaly in patients with liver cirrhosis and portal hypertension is not simply the consequence of portal congestion resulting in decreased SBF. Rather, increased SBF serves to maintain portal blood flow and thereby contributes to portal hypertension. In few patients (15%) SBF increased to more than 11/min may be an important factor for the severity of portal hypertension. Surgical shunt treatment should be adjusted in these patients.


[Extracorporeal shock wave lithotripsy in patients with bile duct calculi]. 21 patients with choledocholithiasis, in whom methods such as endoscopy or mechanical lithotripsy were unsuccessful, extracorporeal shock wave lithotripsy was performed. Fragmentation could be achieved in 90%, complete stone clearance after endoscopic stone extraction occurred in 76%. Extracorporeal shock wave lithotripsy has proven highly effective with a low complication rate and zero lethality. Therefore, its role in the management of gall stone disease in the elderly patient at high perioperative risk cannot be overemphasized.


[Simultaneous sonographic study of postprandial gastric emptying and gallbladder contraction]. Postprandial gastric emptying and gallbladder contraction were assessed in 14 healthy subjects by means of ultrasonography after oral administration of a semi-liquid test meal (250 ml, 1450 kJ). For this purpose, cross-sectional areas of the gastric antrum and gallbladder volume were calculated and recorded over a period of 120 minutes using an annular-array-transducer. The semi-liquid test meal allowed suitable sonographic measurement of cross-sectional areas of the antrum in all 14 subjects. Mean half-time of gastric emptying was 47 minutes (range 17-72 minutes). Mean peak gallbladder contraction was 36% (range 17-60%) of initial volume and mean time to peak contraction was 65 minutes (range 20-120 minutes). The method described was found to be a practical and reliable procedure for the investigation of postprandial gastric emptying and gallbladder contraction. It is therefore of potential interest for application to a variety of clinical questions. A minor drawback involves the relative length and variability of time required for testing, owing to the wide range in the time-course of gastric emptying and gallbladder contraction across subjects. Normal ranges for defined test meals must be established in large control groups.


The importance of MRI (magnetic resonance imaging) for the diagnosis of brainstem infarction. As experience with it accumulates, high resolution MRI seems more capable of representing morphological alterations of the central nervous system than cranial computer tomography (CCT). The question thus arises whether the detection of brainstem infarctions with MRI might not be easier than with CCT and how the topography of brainstem lesions demonstrated with MRI correlate with clinical symptoms and signs. 16 patients with definite clinical symptoms of brainstem infarction were examined by MRI. The examination took place between the 10th and 14th day after the occurrence of the infarction. Using MRI, we found pathological results in only 8 of these cases. In only 5 of these cases a clear relationship between the neurological picture and the topography of the lesion could be shown. These findings demonstrate that despite the high quality of the MRI images, the clinical examination for symptoms and signs is more important than imaging in the brainstem infarction.


Comparison of MRI, CT and angiography in cerebral arteriovenous malformations. Computed tomography (CT), magnetic resonance imaging (MRI) and cerebral angiography were performed in 30 patients with cerebral arteriovenous malformations (AVMs). Clinical data and radiographic findings are presented. The diagnostic values of CT and MRI are compared. 25 of 30 AVMs were seen on CT-scans, 27 on MR-images, 26 by angiography. Typical CT-findings were irregularly hyperdense areas with spotlike contrast enhancement. 4 out of 5 AVMs not depicted by CT were diagnosed for tumor, in one case the malformation was hidden by a hematoma. MRI mostly showed the AVM as round or oval lesions with spongelike structure of low signal intensity in T1-weighted sequences or as single enlarged vessels. The 26 AVM revealed by angiography presented typically enlarged vessels; in 2 cases angiography was without any pathological findings, 2 cases showed signs of a space occupying lesion. The results confirm that MRI is superior to CT in detecting and recognizing AVM. Its strength lies in the possibility to detect abnormal vessels even in presence of fresh or older hematoma, and in the precise information on size and location of the AVM. In our series, MRI and CT-scans together showed 28 of 30 AVMs, the remaining 2 cases were not seen with angiography either but only proven histologically. Although today MR-angiography is able to provide three dimensional images of blood vessels, it cannot yet replace angiography in the exact study of the vessels which is required for surgical planning.


Expansion of the cortical representation of a specific skin field in primary somatosensory cortex by intracortical microstimulation. Intracortical microstimulation (ICMS) was applied to a single site in the middle cortical layers (III-IV) in the koniocortical somatosensory fields of sodium pentobarbital-anesthetized rats (Sml) and new world monkeys (area 3b). Low-threshold cutaneous receptive fields were defined in the cortical region surrounding the stimulation site prior to and following 2-6 hr of 5 microA ICMS stimulation. ICMS stimulation did not usually affect the receptive field location, size, or responsiveness to tactile stimulation of neurons at the stimulation site. However, the number of cortical neurons surrounding the stimulation site with a receptive field that overlapped with the ICMS-site receptive field increased in all studied animals, resulting in an enlarged cortical representation of a restricted skin region spanning several hundred microns. The mean size of receptive fields changed in some but not all cases. These results provide evidence that the responses of cortical neurons are subject to change by the introduction of locally coincident inputs into a single location, and demonstrate a capacity for representational plasticity in the neocortex in the absence of peripheral stimulation. These experimental observations are consistent with hypotheses that the cerebral cortex comprises radially oriented populations of neurons that share a common input, and that these inputs are shaped by coincident activity (see Edelman, 1978, 1987; Merzenich, 1987; Merzenich et al., 1990; von der Malsburg and Singer, 1988).


Characteristics of intracellularly injected infragranular pyramidal neurons in cat primary auditory cortex. Pyramidal neurons in layers V and VI of cat primary auditory cortex (AI) were intracellularly injected with biocytin after functional characterization according to a position relative to an anteroposterior sequence of best-frequency responses. A sample of 19 completely filled neurons was analyzed, and a preliminary classification was made on the basis of dendritic morphology and axon collateral distribution. Layer V cells could be divided into two types. Cells in the upper part of layer V and projecting toward the diencephalon had a large cell body and an apical dendrite with extensive branches in layer I. These cells had few recurrent axon collaterals, and no terminal axonal bushes were formed in the vicinity of the dendritic field. Long horizontal collaterals with many boutons, however, extended in various directions parallel to the cortical surface. By contrast, cells in the lower part of layer V and sending an axon into the putamen, or without an obvious subcortical axon, had a medium soma and an apical dendrite with few branches in layer I. These cells had a dense bush of recurrent collaterals extending into layers II and III and surrounding the dendritic field, but few or no horizontal collaterals. Layer VI injected neurons were more heterogeneous. All had a thin ascending dendrite with oblique branches both ending in layer III. Axon collateral distributions varied from cell to cell. Relatively small cells with an apical dendrite that branched frequently in layers III and IV had a dense network of recurrent collaterals in the dendritic field, but virtually no horizontal collaterals. This type projected toward the diencephalon. Cells with relatively long horizontal collaterals and a weak recurrent system confined to layers V and VI had a unique arborization pattern of basal dendrites. This type may have projected to the claustrum or other cortical areas. One cell with dendritic branches restricted to layer VI had horizontal collaterals predominantly in layer VI. This cell projected into the corpus callosum. The apparent close correlation between extrinsic projections of infragranular neurons and their dendritic morphology and intracortical collateral distributions suggests that differentially projecting cells may engage different elements of intracortical circuitry in AI.


Ideology, history and changing models of liver cirrhosis epidemiology. This analysis describes changes in assumptions about beverage alcohol use as a major cause of liver cirrhosis since the turn of the century. The findings reveal that social and political views shaped by the temperance movement, Repeal era and modern alcoholism movement have had a profound impact on medical and public health interpretations of the role of alcohol use in liver disease. Current clinical and epidemiological approaches that stress the importance of alcohol as a direct etiological agent for liver cirrhosis were popular during the 19th century in the wake of the temperance and prohibition movements, but were greatly modified to attribute a secondary role to alcohol use as a cause of cirrhosis during the Repeal and post-World War II eras. The changes during the Repeal and post-World War II eras coincided with the expansion of alcohol consumption and the liberalization of drinking norms in US society. As a result, the role of alcohol in causing cirrhosis among the general population was minimized and the disease was attributed to industrial toxins and air pollution. Contemporary research has affirmed the importance of beverage alcohol as a causal agent of liver cirrhosis using both clinical and epidemiological studies.


The spatial distribution of pulvinar neurons that project to two subregions of the inferior parietal lobule in the macaque. The distribution of pulvinar neurons that project to the lateral intraparietal area (LIP) and area 7a, two subregions of the inferior parietal lobule in monkeys, was determined using small injections of retrogradely transported fluorescent dyes. Both LIP and 7a received the majority of their thalamic input from contiguous but distinct zones within the medial pulvinar nucleus. Thalamocortical cell bodies that projected to LIP were arranged in a dense, horizontally oriented cluster that was sandwiched between two similar clusters of neurons that projected to 7a. There was minimal overlap of the respective clusters. LIP also received an appreciable input from the dorsal half of the lateral pulvinar nucleus, but tracer placements in 7a resulted in only occasional labeled neurons in the lateral pulvinar.


Classical conditioning and cognitive determinants of subjective craving for opiates: an investigation of their relative contributions. Subjective craving for drugs is frequently reported by detoxified addicts. Explanatory models emphasizing classical conditioning (CC) and cognitive mechanisms have been proposed, and the mechanisms postulated by the two approaches are discussed here. There are three major variants of the CC model, arguing that craving is associated with conditioned changes which are respectively drug-agonistic, opponent-process, or withdrawal reactions. The cognitive model highlights the role of 'outcome expectancies' as determinants of both appetitive and avoidant motivations. The present investigation has identified correlates of craving predicted by the above models, and tested them empirically. Detoxified opiate addicts participated in in a Craving Test requiring them to rate their craving, physical state, and emotions during exposures to both neutral and drug-related material. Other measures, including personality and outcome expectancies for drug use, were also taken, and interrelationships among the variables were explored statistically. Whilst some of the predictions of the CC formulations were borne out, the cognitive model received more consistent support. The theoretical and treatment implications of these data are considered.


Hypercortisolism after opioid discontinuation in rapid detoxification of heroin addicts. Long-term opioid consumption can induce hypoadrenalism through impairment of the hypothalamic-pituitary-adrenal axis. Results of the present study showed that, in heroin addicts, saliva cortisol concentrations varied according to the amount of recently consumed heroin and the time elapsed since the last self-administration. Hypercortisolism was observed either after abrupt withdrawal of heroin or the last dose of methadone. Post-detoxification hypercortisolism was still present on day 16 after the last opioid consumption, whereas it was not observed in abstinent addicts for a mean period of 4 months. During detoxification treatment, mean AUC8-24 cortisol in saliva of clonidine or guanfacine-treated patients was significantly higher than that in methadone-treated patients. It may be hypothesized that elevated cortisol levels may account for untoward effects of adrenergic agonist therapy which, in turn, may represent an added risk factor for relapse during detoxification. Further studies are necessary to correlate the severity of withdrawal symptoms to cortisol levels in opioid addicts detoxified with alpha 2-adrenergic agonist substitution.


Patterns of drug use in Castille and Leon (Spain). A total of 2500 individuals, aged 14-70 years and living in Castille and Leon (Spain), were surveyed in the spring of 1989 with regard to their drug consumption and patterns of use. 'Lifetime' drug users' rates were 30.0% for cannabis, 5.4% for cocaine, 5.2% for amphetamines, 2.4% for psychedelic drugs, 1.5% for opiates, 1.1% for tranquillizers, and 0.4% for inhalants. 'Regular' drug users were more common among cannabis--2.8% of the surveyed--and less frequent among cocaine, opiates and amphetamine users--0.3% of everyone of the substances mentioned above. 'Lifetime' drug users were more common among males than among females, among singles than among married, in those younger (18-29 years of age) and with work problems. The starting age of drug consumption ranged between the average of 15.6 years of age for those who consumed inhalants and 19.5 for those who consumed cocaine. The results allow a better understanding of the pattern of drug use in Castille and Leon (Spain), as well as the 'high' prevalence of drug use in our region.


The relationship between stressful working conditions and high alcohol consumption and severe alcohol problems in an urban general population. The relationship between 15 measures of stressful working conditions and high alcohol consumption (35 g 100% ethanol per day or more for men and 25 g or more for women) was studied, using cross-sectional data from a general population survey of 1344 males and 1494 females; the ages 25-64 years in metropolitan Stockholm in 1984. In a longitudinal component of the study, hospitalization and mortality with alcohol-related diagnosis was assessed during 1984-90, and also the association between previous experience of unemployment and high alcohol consumption. Some of the associations, expressed as age-adjusted odds ratios, were positive and some were negative when high alcohol consumption was the endpoint, but there was a clear variation by sex and social class. Generally the positive associations were stronger among male non-manual employees. Among males, there was a clear association between stressful working conditions and subsequent risk of severe medical alcohol-related problems, but the precision of the estimates was low due to low number of cases. The odds ratio was 6.18 (95% confidence interval 1.86, 20.61) for twisted working positions and 6.74 (95% confidence interval 1.67, 27.19). Previous unemployment among males was associated with increased risk for high alcohol consumption, with an odds ratio of 5.71 (95% CI 1.39, 15.97) among those who had been unemployed more than once, and 1.67 (95% CI 0.76, 3.64) among those who had been unemployed once during the previous 5 years. Those and other increased odds ratios were lower when subjects with an alcohol diagnosis at inpatient care during 1980-84 were excluded in the analyses. On the whole, our findings are not conclusive. The strong, but imprecise associations between stressful working conditions and severe alcohol problems, are however challenging, and warrants further studies, preferably with longitudinal design and repeated measurements of both working condition and alcohol habits.


Smoking habits in relapsed subjects from a smoking cessation trial after one year. Reports of smoking cessation studies often claim that many relapsed subjects reduce their smoking. We investigated the smoking habits of relapsers 1 year after quitting in a smoking cessation trial using nicotine or placebo patches. All 289 participants in that study were summoned to a 1-year follow-up visit--148 (57%) of 259 relapsers attended, as did all 30 sustained abstainers. Fewer than 1% of the subjects had quit spontaneously after the primary relapse. Daily cigarette consumption, standard nicotine yield per cigarette, saliva cotinine concentration, expired carbon monoxide level and two nicotine dependency scales were assessed at entry and at the 1-year follow-up. In five of these six smoking-related characteristics, there was a small but significant mean reduction of 7%-27%. A significant weight gain of 0.5 +/- 2.9 kg (mean +/- SD) was recorded in the relapsers compared with 4.8 +/- 4.2 kg for abstainers (p less than 0.001). It is concluded that smoking habits in relapsers are relatively unchanged, and thus the most important outcome measure in smoking cessation trials is abstinent subjects.


Fractionating language: different neural subsystems with different sensitive periods. Theoretical considerations and psycholinguistic studies have alternatively provided criticism and support for the proposal that semantic and grammatical functions are distinct subprocesses within the language domain. Neurobiological evidence concerning this hypothesis was sought by (1) comparing, in normal adults, event-related brain potentials (ERPs) elicited by words that provide primarily semantic information (open class) and grammatical information (closed class) and (2) comparing the effects of the altered early language experience of congenitally deaf subjects on ERPs to open and closed class words. In normal-hearing adults, the different word types elicited qualitatively different ERPs that were compatible with the hypothesized different roles of the word classes in language processing. In addition, whereas ERP indices of semantic processing were virtually identical in deaf and hearing subjects, those linked to grammatical processes were markedly different in deaf and hearing subjects. The results suggest that nonidentical neural systems with different developmental vulnerabilities mediate these different aspects of language. More generally, these results provide neurobiological support for the distinction between semantic and grammatical functions.


Can carrots be addictive? An extraordinary form of drug dependence. The paper describes three cases of dependence on carotenoids with typical symptoms of irritability and nervousness accompanying their abstinence, with a long-term dependence, and an inability to simply discontinue. Three patients (a man and two women) all being smokers, evaluated this dependence as very similar to that on tobacco. The limitation of further use had the same effect in both cases. The women evaluate the dependence as stronger than that on cigarettes, the man as somewhat weaker. The former patient--a woman--even relapsed, and recently found herself in danger of further relapse. This, however, she nipped in the bud. Laboratory enzyme examinations revealed a disorder in the enzymatic outfit affecting the auto-immunity and the neurovegetative system.


Entropy and cortical activity: information theory and PET findings. Functional segregation requires convergence and divergence of neuroanatomical connections. Furthermore, the nature of functional segregation suggests that (1) signals in convergent afferents are correlated and (2) signals in divergent efferents are uncorrelated. The aim of this article is to show that this arrangement can be predicted mathematically, using information theory and an idealized model of cortical processing. In theory, the existence of bifurcating axons limits the number of independent output channels from any small cortical region, relative to the number of inputs. An information theoretic analysis of this special (high input:output ratio) constraint indicates that the maximal transfer of information between inputs, to a cortical region, and its outputs will occur when (1) extrinsic connectivity to the area is organized such that the entropy of neural activity in afferents is optimally low and (2) connectivity intrinsic to the region is arranged to maximize the entropy measured at the initial segments of projection neurons. Under the constraints of the model, a low entropy is synonymous with high correlations between axonal firing rates (and vice versa). Consequently this antisymmetric arrangement of functional activity in convergent and divergent connections underlying functional segregation is exactly that predicted by the principle of maximum preservation of information, considered in the context of axonal bifurcation. The hypothesis that firing in convergent afferents is correlated (has low entropy) and spatially coherent was tested using positron emission tomographic measurements of cortical synaptic function in man. This hypothesis was confirmed.


Early onset of drinking as a risk factor for lifetime alcohol-related problems. Heavy drinking among students has been a major public health concern over the past decade. A nationally representative 1988 survey on drinking practices and related problems examined the effect of age of of onset of drinking on lifetime alcohol-related problems. Prevalence estimates were obtained for major demographic subgroups of the population. Results and implications are discussed in the context of minimum legal drinking age.


Dynamic properties of salmon calcitonin bound to sodium dodecyl sulfate micelles: a restrained molecular dynamics study from NMR data. We have investigated the conformational behaviour of salmon calcitonin bound to sodium dodecyl sulfate micelles by means of restrained molecular dynamics simulations with both 'static' and time-averaged NMR distance restraints. A more realistic picture of the inherent flexibility of the hormone is obtained when using time averaging. With this approach, long-range NOEs are interpreted better by considering a dynamical exchange among different conformations.


Spatial structure of (34-65)bacterioopsin polypeptide in SDS micelles determined from nuclear magnetic resonance data. The spatial structure of a synthetic 32-residue polypeptide, an analog of the membrane-spanning segment B (residues 34-65) of bacterioopsin of Halobacterium halobium, incorporated into perdeuterated sodium dodecyl sulfate micelles, was determined from 1H NMR data. The structure determination included the following steps: (1) local structure analysis; (2) structure calculations using the distance geometry program DIANA; (3) systematic search for energetically allowed side-chain rotamers consistent with NOESY cross-peak volumes; (4) random generation of peptide conformations in allowed conformational space. The obtained structure has a right-handed alpha-helical region from Lys41 to Leu62 with a kink of 27 degrees at Pro50. The C-cap Gly63 adopts a conformation with phi = 87 +/- 6 degrees, psi = 43 +/- 10 degrees typical to a left-handed helix. The N-terminal part (residues 34-40) is exposed to the aqueous phase and lacks an ordered conformation. The secondary structure of segment B in micelles is consistent with the high-resolution electron cryomicroscopy model of bacteriorhodopsin (Henderson et al. (1990) J. Mol. Biol., 213, 899-929).


A systematic comparison of three structure determination methods from NMR data: dependence upon quality and quantity of data. We have systematically examined how the quality of NMR protein structures depends on (1) the number of NOE distance constraints, (2) their assumed precision, (3) the method of structure calculation and (4) the size of the protein. The test sets of distance constraints have been derived from the crystal structures of crambin (5 kDa) and staphylococcal nuclease (17 kDa). Three methods of structure calculation have been compared: Distance Geometry (DGEOM), Restrained Molecular Dynamics (XPLOR) and the Double Iterated Kalman Filter (DIKF). All three methods can reproduce the general features of the starting structure under all conditions tested. In many instances the apparent precision of the calculated structure (as measured by the RMS dispersion from the average) is greater than its accuracy (as measured by the RMS deviation of the average structure from the starting crystal structure). The global RMS deviations from the reference structures decrease exponentially as the number of constraints is increased, and after using about 30% of all potential constraints, the errors asymptotically approach a limiting value. Increasing the assumed precision of the constraints has the same qualitative effect as increasing the number of constraints. For comparable numbers of constraints/residue, the precision of the calculated structure is less for the larger than for the smaller protein, regardless of the method of calculation. The accuracy of the average structure calculated by Restrained Molecular Dynamics is greater than that of structures obtained by purely geometric methods (DGEOM and DIKF).


Measurement of two-bond JCOH alpha coupling constants in proteins uniformly enriched with 13C. A simple E.COSY type technique is described for measurement of two-bond JCOH alpha coupling constants in proteins that are uniformly enriched with 13C. The method has been used to measure 2JCOH alpha for 132 residues in the proteins calmodulin and staphylococcal nuclease having non-overlapping H alpha-C alpha correlations. Measured 2JCOH alpha coupling constants fall in the 0 to -9.5 Hz range. A separate experiment, measuring the accuracy of these values, indicates a root-mean-square error of 1 Hz. Comparison of the J couplings with the dihedral backbone angles from crystallographic studies confirms a weak but statistically significant correlation between the dihedral angle psi and the magnitude of 2JCOH alpha, but also indicates that parameters other than psi have a significant effect on the value of the coupling.


Intracellular boron localization and uptake in cell cultures using imaging secondary ion mass spectrometry (ion microscopy) for neutron capture therapy for cancer. Quantitative ion microscopy of freeze-fractured, freeze-dried cultured cells is a technique for single cell and subcellular elemental analysis. This review describes the technique and its usefulness in determining the uptake and subcellular distribution of the boron from boron neutron capture therapy drugs.


The imaging and quantification of aluminium in the human brain using dynamic secondary ion mass spectrometry (SIMS). Dynamic secondary ion mass spectrometry (SIMS) has been utilised to study the post-mortem distribution of aluminium in air-dried frozen sections from unfixed, unstained human brain in order to minimise contamination of the tissue and avoid redistribution and extraction of endogenous tissue aluminium. Substrates, sputter-coated with silver, were found to be free of focal aluminum surface contamination and thus minimised substrate induced artefacts in the tissue aluminium ion image. SIMS imaging of aluminium secondary ions at a mass resolution that eliminated the major molecular interferences, combined with a photomontage technique provided a unique strategy for studying aluminium distribution in tissue unrivalled by other spatially resolved microanalytical techniques such as laser microprobe mass spectrometry or X-ray microanalysis. Using this strategy, high densities of focal aluminium accumulations have been demonstrated in the cerebral cortex of the majority of chronic renal dialysis patients studied. In contrast, such aluminium accumulations were absent in control patients. SIMS imaging of aluminium appeared to provide much better discrimination between the dialysis patient group and the control group than one of the most widely used techniques for measuring aluminium in bulk samples, graphite furnace atomic absorption spectrometry. Preliminary studies have shown the feasibility of quantifying focal aluminium SIMS images obtained from brain tissue using aluminium-loaded brain homogenates as reference standards.


Preservation of the diffusible cations for secondary ion mass spectrometry. II. Artefacts in material embedded in araldite or melamine. Flotation on hot water (about 60 degrees C) which is frequently employed to stretch semithin sections on substrates for SIMS (secondary ion mass spectrometry) microscopy, is the cause of numerous artefacts. In the case of epoxy resin-embedded tissue, one observes loss of potassium and sodium and accumulation of calcium. The relative contrast of cell nuclei in the ionic images, is rapidly affected by these ion migrations. After prolonged contact with hot water, tissue becomes uniformly emissive. In the case of hydrosoluble resin-embedded tissue, potassium and sodium do not appear to be affected by the action of water, which suggests that they are covalently bound with chelating sites buried beneath the layer of water bound to the surface of the macromolecules. Calcium accumulates, probably on widely exposed anionic sites. Moreover, the domains observed in hydrosoluble resin-embedded tissue shrink differently according to the proportion of water removed by melamine; this can provide interesting information on the initial equilibrium between water, ion sand macromolecules. Our results seem to support the assumption that bound water should play an important role in the preservation of both macromolecular architecture and ion distributions.


The role of secondary ion mass spectrometry (SIMS) in biological microanalysis: technique comparisons and prospects. The virtues and limitations of SIMS ion microscopy are compared with other spectroscopic techniques applicable to biological microanalysis, with a special emphasis on techniques for elemental localization in biological tissue (electron, X-ray, laser, nuclear, ion microprobes). Principal advantages of SIMS include high detection sensitivity, high depth resolution, isotope specificity, and possibilities for three-dimensional imaging. Current limitations, especially in comparison to X-ray microanalysis, center on lateral spatial resolution and quantification. Recent SIMS instrumentation advances involving field emission liquid metal ion sources and laser post-ionization will help to minimize these limitations in the future. The molecular surface analysis capabilities of static SIMS, especially with the new developments in commercial time-of-flight spectrometers, are promising for application to biomimetic, biomaterials, and biological tissue or cell surfaces. However, the direct microchemical imaging of biomolecules in tissue samples using SIMS will be hindered by limited concentrations, small analytical volumes, and the inefficiencies of converting surface molecules to structurally significant gas phase ions. Indirect detection using elemental or isotopically tagged molecules, however, shows considerable promise for molecular imaging studies using SIMS ion microscopy.


Preservation of the diffusible cations for SIMS microscopy. I. A problem related to the state of water in the cell. The notion of diffusible ions is reviewed in the light of recent knowledge on the stage of water in biological matrices. It appears that ion distributions would be little affected as long as water-macromolecular equilibrium is maintained, but they risk to be significantly modified during dehydration, because the transformation of bound water into highly solvating free water can produce ion displacements. In addition, differently hydrated areas may undergo unequal volume variations. The principal modes of preparing material for SIMS (secondary ion mass spectrometry) microscopy are envisaged from this viewpoint.


Sample preparation of animal tissues and cell cultures for secondary ion mass spectrometry (SIMS) microscopy. Sample preparation is a critical step in the elemental analysis of animal tissues and cell cultures with ion microscopy. Since live cells cannot be analyzed with ion microscopy, a careful sample fixation is necessary which preserves the native structural and chemical integrity of a specimen. The evaluation of morphological and chemical integrity of a fixed specimen is necessary before any physiological explanation of ion fluxes is interpreted based on ion microscopy. For diffusible ion localization studies, strict cryogenic procedures are recommended. Examples are shown for diffusible ion microanalysis in frozen-freeze-dried tissues and cell cultures. Ion microscopy studies of tightly bound elements/molecules may be conducted in chemically fixed and/or plastic embedded specimens. Since it is not generally known which elements/molecules are tightly bound to the tissue matrix, a confirmation of elemental distribution with cryogenic procedures is desirable. A recent approach of combining laser scanning confocal fluorescence microscopy and ion microscopy on the same frozen freeze-dried cell is also discussed for recognizing smaller cytoplasmic structures in ion microscopy images.


Scanning secondary ion analytical microscopy with parallel detection. The secondary ion microscope described here allows to obtain the simultaneous registration of chemical and isotopic distribution maps of several elements composing the sample. The instrument has been specially designed to optimize both sensitivity and selectivity; bombardment with primary Cs+ ions to increase the ionization yields of negative secondary ions, efficient collection of secondary ions at the target surface, matching of the secondary ion beam etendue with the acceptance of the mass spectrometer working at high mass resolution, spectrometer with parallel detection capabilities. The probe diameter can be made as low as 30 nm and ion induced electron images registered at the same time as ion images. Presently, four ion micrographs are obtained simultaneously over a field of view up to 20 x 20 micro m2 containing up to 512 x 512 pixels. Examples are shown with an ion probe diameter of 0.1 microm.


SIMS microscopy in the biomedical field. We attempted to indicate the requirements for biomedical applications of SIMS microscopy. Sample preparation methodology should preserve both the structural and the chemical integrity of the tissue. Furthermore, it is often necessary to correlate ionic and light microscope images. This implies a common methodological approach to sample preparation for both microscopes. The use of low or high mass resolution depends on the elements studied and their concentrations. To improve the acquisition and processing of images, digital imaging systems have to be designed and require both ionic and optical image superimposition. However, the images do not accurately reflect element concentration; a relative quantitative approach is possible by measuring secondary ion beam intensity. Using an internal reference element (carbon) and standard curves the results are expressed in micrograms/mg of tissue. Despite their limited lateral resolution (0.5 microns) the actual SIMS microscopes are very suitable for the resolution of biomedical problems posed by action modes and drug localization in human pathology. SIMS microscopy should provide a new tool for metabolic radiotherapy by facilitating dose evaluation. The advent of high lateral resolution SIMS imaging (less than 0.1 microns) should open up new fields in biomedical investigation.


Cytogenetic applications of high resolution secondary ion imaging microanalysis: detection and mapping of tracer isotopes in human chromosomes. Analytical imaging by secondary ion mass spectrometry (SIMS) using a state-of-the-art scanning ion microprobe enables the detection and mapping of tracer isotopes in human metaphase chromosomes. The stimulated mitosis of cells cultured in media containing labelled nucleosides, typically 14C-labelled thymidine or adenosine, and BrDU, yields chromosomes that have incorporated the labelled molecule in their constituent DNA. The label is subsequently detected and localized by SIMS imaging. The relative label signal intensities of sister chromatids can be quantified. The occurrence of sister chromatid exchanges (SCE) can be detected. The distribution of specific nucleosides can be directly mapped. This is non-uniform along the chromatids, giving rise to characteristic banding patterns (SIMS bands) that seem to correspond to the well known G- or Q-bands resulting from conventional staining methods. The study of a number of cytogenetic problems is expected to benefit from the use of this new method of approach, similar in principle, but potentially more sensitive and capable of higher spatial resolution than autoradiography.


Significance of SIMS microscopy for the radioiodine detection in animal and human thyroid tissue. We defined the SIMS conditions for radioiodine detection in animal and man thyroid follicles, in tissue sections (3 microns) chemically fixed and resin embedded. Two radioisotopes were tested: 125I and 129I, of high (14 mCi 125I micrograms-1) and low specific activity (1.07 10(-6) mCi 129I micrograms-1). In animal study, Wistar rats fed a normal iodine diet (10 micrograms 127I day-1) were injected ip 24 h before sacrifice either with 125I (7 10(-3) micrograms) or with 129I at a dose identical to iodine diet (10 micrograms) or 3 times higher (30 micrograms). No SIMS signal of 125I was obtained in vivo due to its too low concentration, while radioiodine distribution was evidenced with both doses of 129I. Local concentration of previously stored 127I in follicular lumen was not modified, when compared to control (4.14 +/- 0.03 micrograms/mg, m +/- SE), by 125I or 129I at a dose of 10 micrograms, but was nearly doubled with 129I at a dose of 30 micrograms, proof of a pharmacological effect on thyroid iodine regulation. In human study 129I was excluded due to its long half-life (1.6 10(7) years), and 125I was tested only in vitro on two surgical specimens of normal perinodular thyroid tissue maintained in mini-organ culture for 48 h in presence of 100 microCi/ml of 125I. The 125I was detectable, its concentration was 1,000-fold higher than that of 127I (1.5 +/- 0.004 micrograms/mg). For both in vivo and in vitro studies, a positive correlation exists between newly organified radioiodine (125I or 129I) and previously stored iodine (127I).(ABSTRACT TRUNCATED AT 250 WORDS)


Mapping the cellular distribution of labelled molecules by SIMS microscopy. We took advantage of one of the main possibilities of ion microscopy, ie isotopic analysis, to study the cellular distribution of molecules labelled either with carbon 14 or with stable isotopes of low natural abundance such as nitrogen 15 and deuterium. The surface of the sample is bombarded with an ion beam (O2+, Cs+ etc). Secondary ions emitted from the sample are filtered by a mass spectrometer and the distribution of the labelling isotope is recorded. In this way, we obtained images showing the characteristic distribution of 14C-thymidine and D-arginine in human fibroblasts, and of 15N-adenine in organotypic cultures of human breast cancer cells. The spatial resolution on the acquired images was close to 0.1 micron when using the UPS-ONERA ion microprobe. The sensitivity of the method for detecting carbon 14 is far greater than that of autoradiography and the technique is both fast and quantitative. On the other hand, the capacity of ion microscopy for studying the tissular distribution of molecules labelled with stable isotopes, opens the way for biological and pharmacological tracer studies of human diseases.


SIMS microscopy: a tool to measure the intracellular concentration of carbon 14-labelled molecules. Monolayer cultures of human fibroblasts were incubated for 24 h with 14C-arginine and observed by means of SIMS microscopy (ion microscopy). Carbon 14 imaging showed the intracellular distribution of labelled arginine which featured high nuclear incorporation. The local concentration of this amino acid in different cells and intracellular structures was assessed through local isotopic 14C/12C ratio measurement. This relates the signal intensity of the labelling isotope carbon 14 to that of the corresponding natural isotope (carbon 12) of known tissular concentration. Using this method we were able to measure minor variations in the molecular concentration of arginine (expressed as mumol/g of tissue) between different fibroblasts. Results of this study indicate that SIMS microscopy is well adapted to carbon 14 detection and can provide quantitative maps of the cellular and subcellular distribution of 14C-labelled molecules.


Mapping of intracellular halogenous molecules by low and high resolution SIMS microscopy. The subcellular distribution of halogenous molecules has been studied by SIMS microscopy in cultured cells of a human breast carcinoma (MCF-7 cell line). Two instruments of microanalysis were used. A low lateral resolution ion microscope (SMI 300 CAMECA) and a prototype scanning ion microscope equipped with a cesium gun that gives high lateral resolution images. This apparatus has been developed by G Slodzian, in Onera Laboratories (Office National d'Etudes et de Recherches Aérospatiales). Molecules studied by low lateral resolution ion microscope were halogenous steroids: fluorometholone, triamcinolone, bromocriptine and bromoandrosterone. Analytical images show that the first two compounds are mainly localized in the nuclear structure of MCF-7 cells whereas the last two molecules are localized in cytoplasm of these cells. Images were obtained with a resolution of 1 micron. With the scanning ion microscope, it is now possible to obtain images at the ultrastructural level. Four analytical images can be simultaneously obtained by a single scan of the imaged area, corresponding to a depth of erosion of the section of ten nm. The intranuclear distributions of three pyrimidine analogs, 5-bromo-2'-deoxyuridine, 5-iodo-2'-deoxyuridine and 5-fluorouracil have been studied in phase S and M of MCF-7 cells and these images have been compared to the distribution of sulfur, nitrogen and phosphorus. All these images have been obtained with a lateral resolution better than 100 nm.


Subcellular localization of two neurotropic drugs in three varieties of central nervous system cells by secondary ion mass spectroscopy (SIMS) microscopy. The intracellular localization of two neurotropic drugs, flunitrazepam (benzodiazepine) and triflupromazine (phenothiazine), was studied by secondary ion mass spectrometry microscopy (SIMS) in three varieties of cells. The images of the intracellular distributions of the two drugs are easily obtained by selecting the fluorine atom of the molecules. These images show that the drug from the benzodiazepine group is mainly located in the nuclei, whereas the phenothiazine is exclusively located inside the cytoplasm.


Marrow transplantation for Fanconi anemia with or without leukemic transformation: an update of the Seattle experience. Between March 1973 and August 1990, 17 patients with Fanconi anemia (FA) underwent bone marrow transplantation in Seattle. Marrow donors were HLA identical siblings (n = 14), phenotypically HLA identical parents (n = 2) and a one antigen mismatched parent (n = 1). Patients with no evidence of leukemic transformation (n = 12) were conditioned with 140-200 mg/kg cyclophosphamide (CY). Of five patients with leukemic transformation, four received CY (120 mg/kg) plus 12 Gy fractionated total body irradiation and one patient received busulfan (14 mg/kg) and CY (100 mg/kg). All patients engrafted; however, one patient whose sibling donor's cells showed variable results when assayed for chromosome instability required two additional marrow infusions. Toxicity associated with the conditioning regimen included severe oral mucositis (n = 14), hemorrhagic cystitis (n = 11) and diffuse erythroderma (n = 3). Seven of the 12 patients without leukemic transformation are surviving 1-17 years (median = 5 years) after transplant, with an estimated survival probability at 5 years of 65% (95% CI 0.31; 0.85). Two patients developed squamous cell carcinoma of the tongue greater than 10 years post-transplant. One of these patients died at 10.3 years as a result of the malignant process, and the other is disease free more than 12 years post-transplant. Of the five patients with leukemic transformation, one is alive at 8 years. These data demonstrate that marrow transplantation can offer long-term survival for patients with FA, engraftment can be achieved with reduced doses of CY in FA patients, and less toxic preparative regimens are needed for FA patients who have developed leukemic transformation.


Cytokine activity after allogeneic bone marrow transplantation. IV. Production of mRNA for IL-3 and GM-CSF by mitogen-stimulated circulating mononuclear cells. The ability of circulating mononuclear cells from recipients of HLA identical sibling marrow transplants to generate messenger (m) RNA for the haemopoietic growth factors interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) upon mitogen stimulation was investigated. The amount of IL-3 mRNA per ml of blood and IL-3 mRNA per 10(7) mononuclear cells as well as the amount of GM-CSF mRNA per ml of blood was significantly lower in transplant recipients than in normal volunteers. Lower values for mRNA expression for both IL-3 and GM-CSF were associated with the use of immunosuppressive therapy post transplant. No correlation was found between the expression of message for either cytokine, or the presence or absence of acute graft-versus-host disease at the time of testing. While there was no correlation between the quantity of GM-CSF message produced and time elapsed post transplant, IL-3 message expression increased slightly with increasing time post transplant. These defects of haemopoietic regulators constitute another parameter of impaired cellular immunity occurring after allogeneic marrow transplantation.


Sexual satisfaction in survivors of bone marrow transplantation. Of 157 (80%) adult survivors of bone marrow transplantation, 126 responded to a mailed survey 6-149 months after transplant concerning their ability to attain sexual satisfaction. Sixty-five percent of the subjects indicated some degree of sexual satisfaction (with 29% indicating a high degree of satisfaction). Thirteen percent were neutral and 22% were dissatisfied. Gonadal physiological disturbances were present in most subjects as a result of intensive treatments. Fifty-seven percent of women were receiving estrogen replacement, and only 40% had regular menses. Twenty-four percent of men had difficulty with erections and 13% had ejaculatory dysfunction. For men, difficulties with erection (r = 0.54, p less than 0.001) or ejaculation (r = 0.52, p less than 0.001) were associated with inability to attain sexual satisfaction. For women, loss of menses (r = 0.34, p = 0.008) was associated with inability to attain sexual satisfaction. Although few women had abnormally low serum estradiol (even with hormonal supplementation), and none of the 43 men tested had abnormally low serum testosterone, sexual satisfaction correlated with serum levels of estrogen in women (r = 0.42, p = 0.04) and serum testosterone in men (r = 0.28, p = 0.04). Forty-two subjects rated their level of sexual satisfaction before and after transplant. Post-transplant satisfaction was not associated with pretransplant satisfaction (r = 0.07, p = 0.67).(ABSTRACT TRUNCATED AT 250 WORDS)


Relapse of leukemia after bone marrow transplantation: effect of second myeloablative therapy. Twenty-three patients with recurrent leukemia after bone marrow transplantation (BMT) underwent a second myeloablative therapy followed by second transplant between June 1977 and April 1991. Patients had either acute lymphocytic leukemia (n = 4), acute myelogenous leukemia (n = 7) or chronic myelogenous leukemia (n = 12). The median age was 29 years and the median interval between first and second BMT was 22.6 months. The second preparative therapy consisted of cyclophosphamide (200 mg/kg) and total body irradiation (1200 cGy) in nine patients and busulfan (16 mg/kg) and cyclophosphamide (120-200 mg/kg) with or without etoposide (30-60 mg/kg) in 14 patients. The same sibling donor (three syngeneic, 20 allogeneic) was used for both transplants. All patients demonstrated prompt neutrophil recovery (median 21 days) with donor-derived hematopoiesis documented in 16 of 16 evaluable patients. With a median follow-up of 24 months after second BMT, the survival, event-free survival and probability of remaining in remission at 26 months are 47%, 38% and 76% respectively. Outcome was best in patients with chronic myelogenous leukemia (7/12 survivors) and worst in patients with acute leukemia (2/11 survivors). Thus, the data would suggest that (1) selected patients with recurrent leukemia after BMT can still be cured with myeloablative therapy and second BMT, and (2) further improvements in outcome will be dependent upon the reduction of regimen-related toxicity.