Comparative investigation of disposition of 3,4-(methylenedioxy)methamphetamine (MDMA) in the rat and the mouse by a capillary gas chromatography-mass spectrometry assay based on perfluorotributylamine-enhanced ammonia positive ion chemical ionization. A gas chromatography-mass spectrometry assay based on perfluorotributylamine-enhanced ammonia positive ion chemical ionization has been developed for MDMA and three of its primary metabolites in biological specimens; the assay is linear from 2 to 1000 ng ml-1. Quantitatively, more of an administered dose of 10 mg kg-1 MDMA was excreted by the mouse (72%) than by the rat (35%); most in both species was excreted in urine and within 24 h. The difference in per cent excretion is entirely due to proportionally greater excretion of the parent drug by the mouse. 4-Hydroxy-3-methoxymethamphetamine (HMM) is the major urinary metabolite in both species. HMM and another primary metabolite, 4-hydroxy-3-methoxyamphetamine (HMA), were excreted mainly as glucuronide and sulphate conjugates (> 85%).


Papillary thyroid carcinoma: a multivariate analysis of prognostic factors including an evaluation of the p-TNM staging system. To analyse the prognostic factors in papillary thyroid carcinoma, and in particular to evaluate the accuracy of the pathological tumour, nodes, metastases (p-TNM) staging. Retrospective univariate and multivariate analysis. University hospital in Norway. 167 patients who were operated on for papillary thyroid carcinoma between 1971 and 1985. Main outcome measures--Death of papillary thyroid carcinoma, and length of recurrence free survival. Male sex, increasing age, larger tumours, and spread of growth beyond the thyroid all independently increased the risk of dying of papillary thyroid carcinoma, whereas the period of recurrence free survival was influenced only by the presence of regional metastases and the patient's age. The age related p-TNM staging is suitable for predicting the likelihood of death, but is less accurate in the prediction of recurrence free survival. The age of 45 years is too low to be useful in predicting survival, especially in women. The identification of sex in the multivariate analysis as a strong independent predictor of death of papillary thyroid cancer suggests that the prognostic value of the age related p-TNM staging system could be improved if sex was adjusted for, and if a different age was used for men and women.


Herniorrhaphy in patients aged 80 years or more. A prospective analysis of morbidity and mortality. To find out the morbidity and mortality after repair of groin hernias in patients aged 80 years or more, and to identify factors that add to the risk of hernia repair. Prospective open study. All general surgical departments in Ringkøbing County, Denmark. All 39 patients aged 80 years and over who were admitted with hernias during a one year period (1990). Morbidity and mortality. Three patients refused operation, and of the remaining 36, 15 (42%) were admitted as emergencies (5 of whom were already waiting for elective repair of their hernias). The median age was 84 years (range 80-90) and 23 (64%) were men, 31 patients had inguinal hernias, 4 had femoral hernias, and one an obturator hernia. There were six major and two minor complications after 14 emergency operations (57%), and one minor complication after 22 elective operations (5%, p = 0.0007). Two patients died, both after emergency operations (14%). Elective hernia repair can be carried out safely even in the presence of serious coexisting disease, and emergency hernia repair carries a high risk of complications even in the absence of coexisting disease.


Effects of jejunoileal bypass on oxalate and mineral metabolism in rats. To reassess the effects of jejunoileal bypass on the gastrointestinal absorption and bone metabolism of certain minerals in rats, and to see if jejunoileal bypass in rats was a suitable model in which to study formation of calcium oxalate renal stones. Controlled study. Division of Experimental Surgery, University of Erlangen, Germany. 43 male Sprague-Dawley rats. 23 rate underwent jejunoileal bypass, and 20 laparotomy, with transsection and anastomosis of the jejunum and ileotomy and suture (sham operation). Rats that had undergone jejunoileal bypass ate less and gained less weight than those that had had sham operations. Absorption of calcium and phosphorus from the intestine was impaired, but that of magnesium was unchanged. Absorption of oxalate from the small intestine was unchanged, but that from the colon was increased. There were no signs of hyperoxaluria or urolithiasis. Serum mineral homeostasis was not affected by jejunoileal bypass nor were bone volume, density, or mineral concentrations. Serum concentrations of parathyroid hormone and 1,25-dihydroxycholecalciferol remained low, suggesting that jejunoileal bypass might have induced some calcium flux towards the vascular space. Jejunoileal bypass halts weight increase in rats; the model may be helpful in elucidating associations between enteric factors and calciotropic hormones, and several metabolic features that are altered by jejunoileal bypass in man are not altered in rats.


Acute appendicitis in pregnancy: complications and subsequent management. To establish guidelines for the management of a pregnancy that is complicated by acute appendicitis. Retrospective study. University Hospital, Copenhagen, Denmark. 16 patients operated on for symptoms of acute appendicitis during the 15 year period 1974-1988. In 12 patients (75%) the diagnosis was confirmed histologically. The signs and symptoms were classic, and three patients had contractions. One fetus died, in a patient with appendicitis complicated by intraperitoneal abscess. In all uncomplicated cases the pregnancy proceeded to term and the deliveries were normal. Pregnancy should not deter a surgeon from removing an appendix, once the diagnosis is suspected; no pregnancy was affected by removal of a normal appendix. We recommend that prophylactic antibiotics and tocolytic drugs be given in all cases. Simultaneous caesarean section should be done only if there are obstetric indications.


Hemodynamic observation and treatment approach for patients with angina decubitus. In order to investigate the mechanism and treatment of angina decubitus, 20 patients (18 men and 2 women aged 36-70 years) were studied during hospitalization. All patients were found to have an increased heart rate x systolic blood pressure product before the onset of angina decubitus, indicating that this type of angina pectoris belongs to the category of effort angina. Of the 11 patients investigated by continuous hemodynamic monitoring, 3 had significant progressive increases in pulmonary artery systolic pressure (PASP) and pulmonary artery diastolic pressure (PADP) before onset: their episodes of angina could not be completely controlled by digoxin and diuretics, but quickly subsided after beta blockers were added. Among the other 8 patients, PADP increased slightly in 5 and remained unchanged in 3 cases before onset: these patients had no manifestations of LV dysfunction, and beta blockers combined with coronary vasodilators produced satisfactory effects. These results indicate that LV failure is not a major factor in the pathogenesis of angina decubitus. The LV diastolic dysfunction seen in 8/11 cases may have been related to LV hypertrophy caused by long-term hypertension or chronic persistent ischemia.


Colonic volvulus. Diagnosis and results of treatment in 82 patients. To test the accuracy of initial diagnosis of colonic volvulus and the results of different treatment regimens. Retrospective population based study. Tampere University Hospital (major referral center). All patients who presented with colonic volvulus from 1973-1990, 58 patients had sigmoid, 23 caecal and one had transverse colonic volvulus. Findings of endoscopic or operative treatment compared with the clinical diagnosis and plain abdominal radiographs. Association between treatment and risk factors. Diagnosis was difficult, despite some differences in clinical presentation. Gangrenous bowel was diagnosed only at operation, although caecal volvulus with gangrenous bowel was associated with a high white cell count. In 23 patients with caecal volvulus both right hemicolectomy (n = 11) and tube caecostomy (n = 7) were successful with one death after each procedure and no recurrences. In sigmoid volvulus, resection (n = 19) and detorsion with or without sigmoidopexy (n = 21) resulted in similar numbers of complications and deaths (6 and 4, and 5 and 3, respectively), though recurrences were more common after detorsion (1 (5%) compared with 5 (24%)). Endoscopic decompression was tried in 30 and was successful in 26 cases; it was the only treatment in 17/58 patients, with two deaths (12%) and five recurrences (29%). The overall mortality was 15%, but this was associated more with neuropsychiatric diseases, old age, and residence in mental or nursing homes than with gangrene of the bowel. Poor diagnostic accuracy is a problem. Caecal volvulus can be safely treated by resection or tube caecostomy. Sigmoid volvulus is best treated by endoscopic detorsion followed by operation in otherwise fit patients. Mortality is associated with neuropsychiatric diseases and old age.


Regulation of anterior cell-specific mec-3 expression during asymmetric cell division in C. elegans. The homeobox-containing mec-3 gene of C. elegans is expressed in 10 mechanosensory neurons and is necessary for these cells to acquire their fate. All the mec-3-expressing cells are anterior daughters from an asymmetric cell division. In this paper, we examine the expression of a mec-3--lacZ fusion in the presence of mutations that may disrupt asymmetric cell division or anterior-posterior positional information, as well as mutations that may specifically alter mec-3 expression. A mutation in lin-17 causes production of additional mec-3-expressing cells and can have its effect on the cell division that produces a mec-3-expressing cell. In a lin-5 mutant, in which postembryonic blast cells do not complete cell division and become polyploid, blast cells that would give rise to mec-3-expressing daughters instead express mec-3 themselves. In a lin-12 glp-1 double mutant, which is disrupted for many cell interactions in which two cells compete for the same fate, mec-3 expression is unaffected. These results are consistent with a model for asymmetric cell division in which the mec-3-expressing cell and its sister are different immediately upon cell division, rather than acquiring differences through later interaction with each other or their surroundings. lin-17 mutant animals also show defects in the position of the PVM cell and the PLM axons. Animals mutant in unc-73 and unc-40, known to have axon outgrowth defects, also show errors in PVM position and a low frequency of additional mec-3-expressing cells, as well as occasional secondary vulval protrusions, a common phenotype of lin-17 animals. Many other mutations have either no effect on mec-3 expression or an effect that can be largely predicted from previously known phenotypes: these include mab-5, mig-1, unc-53, egl-5, lin-32, and egl-27. unc-11 shows an unexpected and specific defect in mec-3 expression in the PVD neurons, but not in the other mec-3-expressing cells. Two mutations that suppress the egg-laying defect of unc-86 have no effect on the mec-3 expression defect in an unc-86 mutant.


Pericholangitis with ulcerative colitis following autoimmune hepatitis over 12 years. A 35-year-old woman was diagnosed as autoimmune hepatitis 12 years ago by abnormal findings of liver tests including lupus erythematosus (LE) cell phenomenon and liver biopsy. She was admitted in May 1990 with a history of lower gastrointestinal bleeding. Colonoscopy with biopsy and barium enema revealed chronic ulcerative colitis along the entire colon. Since liver tests did not respond well to prednisolone treatment, liver biopsy was again performed and it revealed periductal inflammation with small duct proliferation, a finding compatible with pericholangitis. We herein report this patient who was initially diagnosed as autoimmune hepatitis and thereafter found to be pericholangitis associated with ulcerative colitis.


Dapiprazole: will it affect the standard of care for pupillary dilation? One of the most controversial clinical issues in optometry is the use of "routine" pupillary dilation. Many optometrists are still reluctant to employ routine dilation because of concerns regarding patient inconvenience, angle-closure glaucoma, or systemic side effects. Dapiprazole, a new alpha-adrenergic blocking agent, is now available to reverse the effects of tropicamide- or phenylephrine-induced mydriasis. This article discusses the advantages, clinical uses, limitations, and legal aspects of this mydriatic antagonist in optometric practice.


Drug therapy for ocular allergy.

Drug therapy for ocular allergy. Ocular allergy is a common problem for the primary care practitioner. A range of ocular decongestants, antihistamines, mast cell stabilizers, and corticosteroids are available for use, and practitioners must be familiar with the clinical effectiveness, treatment regimens, and side effects of these drugs. This paper reviews the drugs, both prescription and nonprescription, appropriate for management of ocular allergy and provides guidelines for their use.


Ocular side effects of selected systemic drugs. Numerous systemic drugs produce adverse effects that involve the eye. Pigmentary inclusions of the lids or conjunctivae or both may be caused by a variety of drugs, including amiodarone, chlorpromazine, and gold salts, while conjunctivitis and blepharoconjunctivitis have been associated with isotretinoin, sulfonamides, salicylates, and antineoplastic agents. Dry eye complaints may be caused by antihistamines, beta-receptor blocking agents prescribed for cardiovascular problems, antianxiety agents, and tricyclic antidepressants. Several drugs have been well documented as causes of keratopathies and/or lenticular deposits, including chloroquine and hydroxychloroquine, chlorpromazine, gold salts, systemic corticosteroids, nonsteroidal antiinflammatory drugs, and the antiarrhythmic agent amiodarone. Visual acuity may be decreased by transient changes in refractive error caused by sulfonamides, the antifungal agent metronidazole, thiazide diuretics, and carbonic anhydrase inhibitors. Dilation of the pupil may be caused by anticholinergic drugs, antihistamines, antidepressant agents, and central nervous system stimulants such as cocaine, methylphenidate, and amphetamines. Nystagmus, diplopia, and extraocular muscle palsies have been associated with central nervous system depressants, antihistamines, barbiturates, and elevated blood ethanol concentrations. Intraocular pressure can be elevated in susceptible individuals by long-term use of topical or systemic corticosteroids. Numerous drugs have been associated with retinal toxicity, including chloroquine and hydroxychloroquine, thioridazine, tamoxifen, and talc, which may embolize to the retinal circulation when administered by long-term drug abusers. The antituberculosis agents ethambutol and isoniazid have been implicated as causes of reduced acuity, visual field defects, and disturbances of color vision. Optic neuritis and retrobulbar neuritis may result from the use of chloramphenicol. This paper describes these and other adverse ocular effects that may be encountered when examining patients who are taking systemic drugs.


Measurement of thyroid-stimulating immunoglobulins by incorporation of tritiated-adenine into intact FRTL-5 cells: a viable alternative to radioimmunoassay for the measurement of cAMP. To evaluate the utility of 3H-adenine incorporation into intact rat thyroid epithelial cells (FRTL-5) as an alternative to radioimmunoassay for the measurement of cAMP following stimulation of these cells by serum thyroid-stimulating immunoglobulins from patients with Graves' disease. We determined the cAMP produced by FRTL-5 cells following incubation with serum from patients with a spectrum of autoimmune thyroid and other diseases using the 3H-adenine assay. We studied 27 patients with untreated Graves' disease, 10 with Graves' disease complicated by ophthalmopathy (all on antithyroid medication), 11 with Hashimoto's thyroiditis, five with multinodular goitre, one with thyroid carcinoma, 23 with type 1 diabetes mellitus, 19 with other autoimmune diseases and 10 controls. The 3H-cAMP produced in cells incubated with either bovine TSH (bTSH) or polyethylene glycol-precipitated serum immunoglobulins, was separated by sequential chromatography on Dowex and alumina columns, and counted. The thyroid-stimulating immunoglobulins index (3H-cAMP patient immunoglobulins/3H-cAMP control immunoglobulins) was calculated for each serum and considered positive if greater than 1.5 (+ve thyroid-stimulating immunoglobulins index, i.e. > 2 standard deviations above control). The thyroid-stimulating immunoglobulins index was correlated with measurement of thyrotrophin binding inhibitory immunoglobulins (TBII) by radioreceptor assay. The 3H-adenine assay has a sensitivity of 10(-11) M bTSH with maximal stimulation at 10(-9) M bTSH (30-fold). Twenty-five of 27 patients (92%) with untreated Graves' disease and four of 10 patients with Graves' disease complicated by ophthalmopathy had +ve thyroid-stimulating immunoglobulin indices. The thyroid-stimulating immunoglobulins index in patients with untreated Graves' disease correlated with their TBII assay result (r = 0.63, P < 0.001). In addition, the index was negative in patients with Hashimoto's thyroiditis, multinodular goitre, thyroid carcinoma, and type 1 diabetes mellitus. Of the patients with other autoimmune diseases only one (a patient with systemic lupus erythematosis) had a +ve thyroid-stimulating immunoglobulin index. Direct comparison of cAMP measurement by 3H-adenine incorporation and commercial radioimmunoassay showed an equal sensitivity to both bTSH and Graves' immunoglobulins. After cell preparation, results are obtained more quickly with the 3H-adenine assay than with a cAMP radioimmunoassay (5 hours compared to 2 days), and far more cheaply than by commercial radioimmunoassays. Measurement of thyroid-stimulating immunoglobulins using the incorporation of 3H-adenine into cAMP in FRTL-5 cells is sensitive, reproducible, rapid and specific. These features make this assay a viable alternative to RIA for the measurement of thyroid-stimulating immunoglobulins in patients with Graves's disease.


Why we have (only) five fingers per hand: hox genes and the evolution of paired limbs. Limb development has long been a model system for studying vertebrate pattern formation. The advent of molecular biology has allowed the identification of some of the key genes that regulate limb morphogenesis. One important class of such genes are the homeobox-containing, or Hox genes. Understanding of the roles these genes play in development additionally provides insights into the evolution of limb pattern. Hox gene expression patterns divide the embryonic limb bud into five sectors along the anterior/posterior axis. The expression of specific Hox genes in each domain specifies the developmental fate of that region. Because there are only five distinct Hox-encoded domains across the limb bud there is a developmental constraint prohibiting the evolution of more than five different types of digits. The expression patterns of Hox genes in modern embryonic limb buds also gives clues to the shape of the ancestral fin field from which the limb evolved, hence elucidating the evolution of the tetrapod limb.


Clox, a mammalian homeobox gene related to Drosophila cut, encodes DNA-binding regulatory proteins differentially expressed during development. We report the isolation of a cDNA encoding a mammalian homeoprotein related to the Drosophila cut gene product, called Clox, for Cut like homeobox. In addition to the homeodomain, three 73-amino acid repeats, the so-called cut repeats, are also conserved between Cut and the mammalian counterpart described here. This conservation suggests that the cut repeat motif may define a new class of homeoproteins. Both cloned and endogenous Clox proteins are nuclear DNA-binding proteins with very similar sequence specificity. Western blot analysis revealed several distinct Clox protein species in a variety of tissues and cell types. The relative abundance of these proteins is regulated during mouse development and cell differentiation in culture. Interestingly, approximately 180-190 x 10(3) M(r) Clox proteins predominate in early embryos and are upregulated in committed myoblasts and chondrocytes, but downregulated upon terminal differentiation. Clox DNA-binding activity is correlated with the abundance of these proteins. In contrast, larger Clox protein species (approximately 230-250 x 10(3) M(r)) are detected mainly in adult tissues and in terminally differentiated cells. Cotransfection experiments show that Clox proteins can function as repressors of tissue-specific gene transcription. Thus, Clox, like their Drosophila counterparts, are candidate regulators of cell-fate specification in diverse differentiation programs.


Identifying targets of the rough homeobox gene of Drosophila: evidence that rhomboid functions in eye development. In order to identify potential target genes of the rough homeodomain protein, which is known to specify some aspects of the R2/R5 photoreceptor subtype in the Drosophila eye, we have carried out a search for enhancer trap lines whose expression is rough-dependent. We crossed 101 enhancer traps that are expressed in the developing eye into a rough mutant background, and have identified seven lines that have altered expression patterns. One of these putative rough target genes is rhomboid, a gene known to be required for dorsoventral patterning and development of some of the nervous system in the embryo. We have examined the role of rhomboid in eye development and find that, while mutant clones have only a subtle phenotype, ectopic expression of the gene causes the non-neuronal mystery cells to be transformed into photoreceptors. We propose that rhomboid is a part of a partially redundant network of genes that specify photoreceptor cell fate.


Long-term effects of befunolol on the corneal endothelium and the consensual ophthalmotonic reaction. In a double-blind study, 30 patients with ocular hypertension were randomly assigned to receive one drop of 0.5% befunolol or placebo (the drug vehicle) in each eye twice daily for 120 days. The befunolol used contained no preservatives. Before and after treatment the number and shape of the endothelial cells of the five corneal sectors were evaluated. After 120 days of twice-daily application of befunolol or placebo, no changes in the shape or density of the corneal cells were found in any patient. No differences in the patients receiving befunolol or placebo were noted. The consensual ophthalmotonic reaction was studied in 15 of the patients before and after befunolol administration. A significant decrease in intraocular pressure was noted at 30 minutes and at 1, 2, 3, 4, 5, and 6 hours after befunolol. Intraocular pressure also decreased significantly at 1 to 4 hours after placebo, and then returned to pretreatment levels at 6 hours.


Exogenous retinoic acid rapidly induces anterior ectopic expression of murine Hox-2 genes in vivo. Exogenous retinoic acid (RA) has teratogenic effects on vertebrate embryos and alters Hox-C gene expression in vivo and in vitro. We wish to examine whether RA has a role in the normal regulation of Hox-C genes, and whether altered Hox-C gene expression in response to RA leads to abnormal morphology. The expression of 3' Hox-2 genes (Hox-2.9, Hox-2.8, Hox-2.6 and Hox-2.1) and a 5' gene (Hox-2.5) were examined by whole-mount in situ hybridization on embryos 4 hours after maternal administration of teratogenic doses of RA on embryonic day 7 to 9. The expression of the 3' Hox-2 genes was found to be ectopically induced in anterior regions in a stage-specific manner. The Hox-2.9 and Hox-2.8 genes were induced anteriorly in the neurectoderm in response to RA on day 7 but not at later stages. Expression of Hox-2.6 and Hox-2.1 was ectopically induced anteriorly in neurectoderm in response to RA on day 8. Hox-2.1 remained responsive on day 9, whereas Hox-2.6 was no longer responsive at this stage. The expression of the 5' gene Hox-2.5 was not detectably altered at any of these stages by RA treatments. We also examined the response of other genes whose expression is spatially regulated in early embryos. The expression of En-2 and Wnt-7b was not detectably altered by RA, whereas RAR beta expression was induced anteriorly by RA on day 7 and 8. Krox-20 expression was reduced in a stage- and region-specific manner by RA. The ectopic anterior expression of Hox-2.8 and Hox-2.9 induced by RA on day 7 was persistent to day 8, as was the altered expression of Krox-20. The altered pattern of expression of these genes in response to RA treatment on day 7 may be indicative of a transformation of anterior hindbrain to posterior hindbrain, specifically, a transformation of rhombomeres 1 to 3 towards rhombomere 4 identity with an anterior expansion of rhombomere 5. The ectopic expression of the 3' Hox-2 genes in response to RA is consistent with a role for these genes in mediating the teratogenic effects of RA; the rapid response of the Hox-C genes to RA is consistent with a role for endogenous RA in refining 3' Hox-C gene expression boundaries early in development.


Induction of labial expression in the Drosophila endoderm: response elements for dpp signalling and for autoregulation. Extracellular signal proteins induce the homeotic gene labial (lab) to high levels of localised expression in the endoderm of Drosophila embryos. We aimed to identify cis-regulatory elements within the lab gene that respond to this induction by analysing the activity of stably integrated reporter gene constructs. Dissection of lab 5' flanking sequences reveals two types of response elements. One of these mediates lab dependent activity, providing evidence that lab induction in the endoderm is autoregulatory. The other element, to a large extent independent of lab function, responds to decapentaplegic (dpp), a signal molecule related to mammalian TGF-beta. Our evidence suggests that lab induction in the endoderm reflects coordinate action of two distinct factors one of which may be lab protein itself, and another whose localised activity or expression in the midgut depends on the dpp signal.


Cell signals allow the expression of a pre-existent neural pattern in C. elegans. In C. elegans, a simple pattern develops within a row of epidermal precursor cells, V1-V6. One cell, V5, gives rise to a neuroblast called the postdeirid neuroblast, while the other V cells produce epidermal cells instead. Here we describe experiments suggesting that in order for V5 to produce the postdeirid it must be in close or direct contact with neighboring V cells. Signaling between V cells is required for the formation of the neuroblast; however, which of the V cells can make a postdeirid is not determined by these signals but rather by the action of the homeotic lin-22 and pal-1 genes. These genes prevent V cells in specific body regions from responding to intercellular signals and producing postdeirids. This is a clear example of cell signals playing a permissive rather than an instructive role in neuroblast induction.


Expression of the homeotic gene mab-5 during Caenorhabditis elegans embryogenesis. mab-5 is a member of a complex of homeobox-containing genes evolutionarily related to the Antennapedia and bithorax complexes of Drosophila melanogaster. Like the homeotic genes in Drosophila, mab-5 is required in a particular region along the anterior-posterior body axis, and acts during postembryonic development to give cells in this region their characteristic identities. We have used a mab-5-lacZ fusion integrated into the C. elegans genome to study the posterior-specific expression of mab-5 during embryogenesis. The mab-5-lacZ fusion was expressed in the posterior of the embryo by 180 minutes after the first cleavage, indicating that the mechanisms responsible for the position-specific expression of mab-5-lacZ act at a relatively early stage of embryogenesis. In embryos homozygous for mutations in the par genes, which disrupt segregation of factors during early cleavages, expression of mab-5-lacZ was no longer localized to the posterior. This suggests that posterior-specific expression of mab-5 depends on the appropriate segregation of developmental factors during early embryogenesis. After extrusion of any blastomere of the four-cell embryo, descendants of the remaining three cells could still express the mab-5-lacZ fusion. In these partial embryos, however, the fusion was often expressed in cells scattered throughout the embryo, suggesting that cell-cell interactions and/or proper positioning of early blastomeres are required for mab-5 expression to be localized to the posterior.


Sequence and embryonic expression of the murine Hox-3.5 gene. The murine Hox-3.5 gene has been mapped and linked genomically to a position 18 kb 3' of its most 5' locus neighbour, Hox-3.4, on chromosome 15. The sequence of the Hox-3.5 cDNA, together with the position of the gene within the locus, show it to be a paralogue of Hox-2.6, Hox-1.4 and Hox-4.2. The patterns of embryonic expression for the Hox-3.5 gene are examined in terms of three rules, proposed to relate a Hox gene's expression pattern to its position within the locus. The anterior boundaries of Hox-3.5 expression in the hindbrain and prevertebral column lie anterior to those of Hox-3.4 and all other, more 5'-located Hox-3 genes. Within the hindbrain, the Hox-3.5 boundary is seen to lie posterior to that of its paralogue, Hox-2.6, by a distance equal to about the length of one rhombomere. Patterns of Hox-3.5 expression within the oesophagus and spinal cord, but not the testis, are similar to those of other Hox-3 genes, Hox-3.3 and Hox-3.4.


Review of the Fogarty International Center Conference, NIH: peptides in health and disease. Taken together, the lectures presented at this conference have amply demonstrated the ever growing impact of peptide research on health, in part by increasing our understanding of disease. This Fogarty International Conference has been invaluable because of the diverse backgrounds of the distinguished participants which it attracted. The lectures and the ensuing discussions justify the greatest expectations for the contribution which peptide research will make to human and animal health during the remainder of this century and beyond.


Secondary prevention of coronary heart disease in patients after acute myocardial infarction at the Soroka Medical Center. The records of 214 patients with acute myocardial infarction (AMI) admitted to Soroka Medical Center in 1989 were reviewed to assess the quality of medical treatment after AMI. Medical treatment was reviewed with respect to administration, indications, and contraindications of specific drugs. Sixty-nine percent of patients received suboptimal treatment with respect to aspirin and beta blockers, which was almost always due to undertreatment rather than overprescription. Only 23% of patients with mural thrombi, as shown by echocardiography and without contraindications to anticoagulants, received coumadin. Patients hospitalized in the Intensive Coronary Care Unit received somewhat better medical care with regard to aspirin, beta blockers and coumadin. Appropriate decisions whether to give or withhold thrombolytic therapy were made in 81% of the cases and it was given to 29% of the patients. We conclude that during the period of this study there was a considerable gap between: a) recommendations in the medical literature, and b) the quality of care provided to AMI patients with regard to medications prescribed by physicians for preventive medical therapy.


[Hormones and the levels of humoral regulation]. The problems on the place of hormones secreted by "classical" endocrine glands, on their relationship with other compounds that possess physiological activity, criteria that determine the definition "hormone" are considered in this article. The conception about the levels of the humoral regulatory systems that are organized and formed during phylogenesis and ontogenesis and provide a consecutive increase in their complexity and mobility of adaptation to changes of environment and internal conditions are substantiated on the basis of numerous data. The metabolites that are products of nonspecific activity of any cell of the multicellular organism form the first and simplest level of humoral regulatory organization. The next (second) level of humoral organization is also formed by chemically simple substances. However, these substances are specialized products of the secretory activity of cells and exert potent influence on the physiological processes. Neuroamines and regulatory peptides are applied to these agents, in the first place. They arise simultaneously and jointly at the first stage of ontogenesis. The distinctive characters of the third level of the humoral regulation are increased and complication of the regulatory activity conditioned by cooperative influences of humoral agents produced by single secretory elements situated outside the classical endocrine glands. The chemically and originally different substances causing predominantly local effects are attributed to these physiologically active substances. Their participation in general adaptive reactions as well as inclusion of classical hormones into hierarchy of humoral regulation signify the formation of the forth regulatory level that provides realization of general homeostatic reactions peculiar to the whole organism.(ABSTRACT TRUNCATED AT 250 WORDS)


A rapid and sensitive method for HLA-DRB1 typing by acridinium-ester-labeled DNA probes. A rapid and accurate detection of HLA class II antigen is essential for transplantation and for the understanding of disease susceptibility. Recent molecular genetic studies have revealed that the number of class II loci and the number of alleles at these loci are greater than had been previously detected. It is, therefore, of great importance to detect these extensive polymorphisms. A great deal of effort has been made on identification of individual HLA class II specificities at the DNA level, called "DNA typing." What seems to be lacking, however, is handling simplicity. Here we accomplished a simple method for HLA-DRB1 typing based on hybridization of acridinium-ester (AE)-labeled DNA probes to amplified DNA. This method is called hybridization protection assay (HPA). By using 13 AE-labeled probes, 20 homozygous B-cell lines and leukocytes from 80 healthy individuals were typed by HPA. The results were completely consistent with those obtained by polymerase chain reaction--restriction fragment length polymorphism. This method is suitable for mass screening because of its procedural simplicity and swiftness.


Peripheral blood stem cell mobilization by cytokines. Hematopoietic stem cells circulate in the peripheral blood. These cells can be collected by apheresis techniques either in the unperturbed state, after mobilization following the administration of cytokines like G-CSF or GM-CSF, or during the phase of early blood count recovery following chemotherapy-induced myelosuppression. The number of cells collected following mobilization is greater than that obtained after apheresis in the unperturbed state. There are, however, qualitative differences between unperturbed and mobilized cells. Chemotherapy related mobilization can be potentially dangerous in that severe myelosuppression necessary to achieve mobilization can have serious consequences. There are no controlled studies that evaluate the relative merits of each method of collection. Regardless of the techniques employed peripheral blood stem cells can reliably accelerate hematologic recovery after potentially myeloblative therapy and provide an alternative to bone marrow support.


Hematopoietic stem cell processing and cryopreservation. Either bone marrow or peripheral blood may be harvested to provide hematopoietic stem cells (HSC) for autologous transplantation. Both, however, comprise heterogeneous cell populations. The HSC necessary for successful engraftment constitute a very small fraction of the cells harvested. After collection, the harvested cells usually undergo several processing steps to reduce the product volume, remove cells (such as mature blood cells or tumor cells), or to cryopreserve the cells for later reinfusion. Granulocytes and red blood cells, for example, survive cryopreservation poorly using freezing techniques designed for HSC. Therefore, bone marrows being cryopreserved must be depleted of mature blood cells to avoid toxicity from infusion of damaged mature blood cells. Mature blood cells may also impede the variety of tumor cell purging techniques currently being studied. These processings are designed to minimize the loss of HSC while achieving an appropriate HSC product for the individual patient. A number of apheresis devices and cell washers simplify the enrichment of HSC in the harvested cell products. In contrast, tumor cell purging techniques are not standardized between the various transplant centers.


Quality assurance and standards in hematopoietic progenitor processing. Bone marrow transplantation is an increasingly important therapeutic procedure. As more laboratories have become involved in the processing of hematopoietic progenitor cells from marrow or blood, it has been recognized that standards are required for hematopoietic progenitor processing, storage, and handling. Quality assurance is the process of monitoring whether laboratory procedures, equipment, and personnel fulfill their expected functions, and the aim of quality assurance is to ensure compliance with standards. Some standards for hematopoietic progenitor processing have recently been issued by professional organizations. Although these standards are not comprehensive, where applicable they should be met or exceeded. In the absence of published standards, principles of good laboratory practice should guide quality assurance programs. This article presents concepts of quality assurance in hematopoietic progenitor processing, based on standard laboratory practice and published standards.


Genetic and immunologic analysis of a family containing five patients with common-variable immune deficiency or selective IgA deficiency. A family with 13 members included 2 subjects with selective IgA deficiency (IgA-D) and 3 subjects with common-variable immune deficiency (CVID), diseases which usually occur sporadically. Reciprocal combinations of B and T cells in vitro between one normal and two immune-deficient family members and normal subjects revealed that defective Ig synthesis was determined by the B cells, while the patient T cells functioned normally. Normal T helper and suppressor function was demonstrated even in one patient with CVID who developed a T-cell lymphoproliferative disorder associated with elevated IgM; this patient's B cells made only IgM in vitro. Immune deficiencies were inherited in this family in a pattern consistent with an autosomal dominant trait with incomplete penetrance. All the immune-deficient patients in this family possessed at least one copy of an MHC haplotype previously shown to be abnormally frequent in IgA-D and CVID: HLA-DQB1*0201, HLA-DR3, C4B-Sf, C4A-deleted, G11-15, Bf-0.4, C2-a, HSP70-7.5, TNF alpha-5, HLA-B8, and HLA-A1. The patient who developed the lymphoproliferative disorder was homozygous for this haplotype. Four immunologically normal members, one of whom was 80 years old, also possessed this MHC haplotype, indicating that its presence is not sufficient for disease expression. A small segment of another MHC haplotype associated with Ig deficiency in the population also occurred in this family, but it was not associated with immune deficiency.(ABSTRACT TRUNCATED AT 250 WORDS)


Preferential expression and activity of multidrug resistance gene 1 product (P-glycoprotein), a functionally active efflux pump, in human CD8+ T cells: a role in cytotoxic effector function. The multidrug resistance gene 1 (mdr 1) product, the P-glycoprotein (Pgp), is a 170-kD transmembrane transport protein, whose overexpression is associated with multidrug resistance in cancer cells and in chloroquine-resistant Plasmodium falciparum infection. In this study we show that normal freshly isolated human lymphocytes express low levels of mdr 1 mRNA and membrane Pgp. Although Pgp is expressed in both CD4+ and CD8+ T cells, it is preferentially expressed in CD8+ T cells. Activation of T lymphocytes with phytohemagglutinin leads to an amplification of both mdr 1 mRNA and membrane Pgp in T cells. P-glycoprotein in T cells is a functionally active efflux pump as demonstrated by decreased retention of rhodamine-123 and its increased accumulation by cyclosporin A, an inhibitor of Pgp function. In addition, MRK-16 antibody increased accumulation of Rh123 in CD8+ T cells. Furthermore, MRK16 anti-P-glycoprotein monoclonal antibody, in a concentration-dependent manner, inhibited T lymphocyte-mediated cytotoxicity. These data suggest a physiologic role of P-glycoprotein in cytotoxic T-lymphocyte effector function.


Endophthalmitis at the Bristol Eye Hospital: an 11-year review of 47 patients. We reviewed data from 47 patients who were treated for endophthalmitis at our hospital during the 11-year period 1980-90. The most common clinical features were hypopyon (75%), diminished vision (72%), ocular pain (68%), discharge (57%), corneal oedema (51%), conjunctival injection (49%), abnormal red reflex (34%), corneal ulcer (32%) and corneal perforation (6%). A total of 54 isolates were obtained from 41 (87%) of the 47 patients. Gram-positive bacteria were more common (72%), than Gram-negative organisms (22%). Two cases were due to fungi, and herpes simplex virus was isolated from one case. The two most common Gram-positive organisms were coagulase-negative staphylococci (25%), and Staphylococcus aureus (11%), while Pseudomonas aeruginosa predominated among the Gram-negative bacteria isolated (15%). Mixed bacterial species were obtained from 29% of the infected patients, including one from whom Vibrio fluvialis was isolated. Predisposing factors included ocular surgery (60%)--mostly for cataract extraction (47%), penetrating trauma (15%) and periocular (15%) or systemic (11%) infections. All patients received antibiotics (generally chloramphenicol and/or a beta-lactamase-stable penicillin plus an aminoglycoside) prior to culture, when treatment was adjusted according to specific aetiological agents. Seventy-nine per cent of patients received topical or systemic steroids. Vitrectomy (diagnostic and therapeutic) was performed on 21% of patients. Sixty-three per cent of culture-positive patients lost vision (no perception of light) in the affected eye, compared to 17% of culture-negative cases (P < 0.05 Fisher exact test). Similarly, a better visual outcome (acuity of 6/12 or better) was associated with coagulase-negative staphylococcal infection than with streptococcal or fungal infections.(ABSTRACT TRUNCATED AT 250 WORDS)


The mechanisms and risks of surgical glove perforation. Intact surgical gloves are a barrier to hepatitis B virus and human immunodeficiency virus (HIV) but once perforated during surgery they cannot sustain adequate defence. This study examines the rate of glove perforations during surgery at a District General Hospital. In total, 275 pairs of gloves were collected from 100 consecutive operations. In the 43% of gloves that had been damaged 200 perforations were recorded. The mean rate per operation in the surgeon's gloves was 1.18. Injuries to the non-dominant index finger were significantly higher than injuries to other parts of the hand. Injuries occurred particularly during manipulation of the needles and at wound closure. Consultants were more likely to have glove perforation than juniors. Operations requiring manipulation of instruments deep within the wound had a higher rate than those on the surface. The results of the study indicate that a surgeon risks more than one hepatitis B infection per lifetime and that at least one in 1500 surgeons is likely to be infected by HIV during the next 35 years.


Colonization with coagulase-negative staphylococci in two neonatal units. Episodes of septicaemia due to coagulase-negative staphylococci (CNS) were more frequent in a level III than in a level II neonatal unit in Stockholm, Sweden. Colonization with CNS during the first 2 weeks of life was investigated in 10 infants from each unit. As the use of antibiotics differed between the two units, the aim was to correlate colonization and antimicrobial resistance patterns to antibiotic usage. Antimicrobial susceptibility of CNS to isoxazolylpenicillins, co-trimoxazole, erythromycin, clindamycin, chloramphenicol and gentamicin was determined. Selected isolates were typed with restriction endonuclease analysis of plasmid DNA and of genomic DNA. Infants were frequently colonized with multiple strains and species of CNS, and transmission of strains from patient to patient occurred within the unit. Qualitative and quantitative differences in antibiotic use were not correlated with colonization. The prevalence of resistant isolates, mostly of Staphylococcus haemolyticus, was higher in the level II unit with lower use of antibiotics. Staphylococcus epidermidis, which is generally more virulent, prevailed in the level III unit, where there were more severely ill children and invasive procedures were more frequently performed.


An outbreak of infection due to Staphylococcus aureus phage type 52 in a neonatal intensive care unit. An outbreak of infection due to Staphylococcus aureus phage-type 52, resistant only to penicillin, occurred in children's wards primarily in a neonatal intensive care unit. The outbreak involved 86 infants; the majority presented with conjunctivitis, wound infections, boils, omphalitis and otitis externa. Seven per cent of these infants (six of 86) also had septicaemia. In addition, 6% (five of 86) were colonized by this phage type and the most common site of carriage was the umbilicus. The outbreak was contained by eradication of nasal carriage among the staff members and also by standard infection control measures.


Bacteraemia caused by non-aeruginosa Pseudomonas species in a cancer centre. Fifty-one episodes of bacteraemia due to Pseudomonas species other than Pseudomonas aeruginosa occurring between 1980 and 1990 in a Belgian cancer centre were reviewed. This corresponded to an incidence of 0.62/1000 admissions, or 1.5% of all bacteraemic episodes. Twenty-nine episodes, each with several positive blood culture sets were considered clinically significant, including six patients belonging to a well-documented outbreak of pseudobacteraemia with Xanthomonas maltophilia and associated with contaminated blood sampling tubes. The respiratory tract was the source in six (20.7%), an infected intravenous catheter in 10 (34.5%) and the source was unknown in seven (24.1%). Seven patients died from infection (24.1%). Twenty-three episodes with a single positive blood culture set were considered clinically not significant, although four of them were considered significant by the Centers for Disease Control (CDC) criteria because of the presence of symptoms and specific antibiotic treatment being administered. None of the patients with a single isolate died from infection despite the fact that 17 of 22 did not receive an effective antimicrobial agent. All isolates were susceptible to co-trimoxazole.


Asymptomatic Salmonella senftenberg carriage in a neonatal ward. During a 23-day period in April 1991, nine infants in a neonatal ward were found to be colonized with Salmonella senftenberg. All were asymptomatic on detection and all except one (who developed septic ileus) remained asymptomatic on follow-up. The affected babies were isolated and subsequently discharged from the ward as soon as possible. These measures, along with emphasis on handwashing and intensification of cleaning and disinfection, were able to prevent spread to other babies. Despite extensive sampling of the environment, mothers and staff, the source of the organism could not be identified.


Efficient and reproducible new semimicromethod for the detection and titration of HIV in human plasma. A semimicromethod was established for isolating human immunodeficiency virus (HIV) in plasma using 48-well plates and a pool of peripheral blood mononuclear cells (PBMC) from several donors as targets for infection, which increases the efficiency of isolation by reducing the effect of variability due to diverse donor cell susceptibility to HIV infection. The addition of H9 cells to the PBMC cultures did not affect measurable titers. Nevertheless, it potentiated strongly virus replication in terms of p24 production in the supernatant of the wells with HIV isolates, thus facilitating interpretation of the results. The titration of a virus strain of a known titre and reverse transcriptase activity in parallel provided a constant parameter of efficiency and reproducibility within each experiment, permitting comparison with results from other laboratories. The reproducibility of the method was highly significant (r = 0.97, P < 0.001); 68% of the 22 plasma samples from HIV-infected individuals tested by this method were positive. The presence of plasma HIV titer correlated well (P < 0.02) with the low count of CD4+ cells of less than 300/mm3, but not with the presence of the p24 antigen in the serum.


Increases in urinary enzyme excretion in rats depleted of glutathione inhibited by scavenger of oxygen free radicals. Urinary excretion of enzymes by rats was assessed after glutathione (GSH) was depleted by treatment with a mixture of the GSH depletors D,L-buthionine-S,R-sulfoximine (BSO) and diethylmaleate (DEM). Renal GSH was low 2 h after treatment and later returned to the control level. The urinary excretion of gamma-glutamyltranspeptidase (gamma-GTP) and N-acetyl-beta-D-glucosaminidase (NAG) remained high for at least 3 d after the injection of BSO (100 mg/kg) and DEM (0.5 ml/kg), with no effect on the blood urea nitrogen level. N,N'-Dimethylthiourea (DMTU), a scavenger of oxygen free radicals, inhibited this increase in the urinary excretion of gamma-GTP. DMTU also inhibited the increase in cisplatin-induced NAG excretion caused by the GSH depletors. These results suggested that the urinary excretion of these enzymes is an index of renal tubular injury caused by short-term depletion of renal GSH, and that the generation of free radicals may be involved in renal tubular injury during GSH depletion or caused by cisplatin together with GSH depletors.


[Irritable bowel syndrome in adolescence]. We studied seventy patients, 23 males and 47 females with irritable bowel syndrome in adolescence aged 13-19 yrs, who visited the department of psychosomatic medicine in Takano Hospital during about six year period of April, 1986-July, 1992. Takano Hospital is a coloproctological center in Kumamoto. In the clinical pattern of adolescent patients with irritable bowel syndrome the "gas" pattern was dominant (51.4%). Patients with the gas pattern have severe symptoms of flatus, fullness, rumbling sound and abdominal pain as well as bowel dysfunction, constipation and diarrhea in a classroom. Next, the diarrheal pattern occurred in 20.0%. Diarrheal patients complained of frequent bowel movements and retention feelings before attending school. Recurrent abdominal pain-like pattern was found in 7.1% patients. Clinical symptoms in the adolescent patients seem to derived from a mental tension and stress in a close classroom or before attending school. Many adolescenct patients (67.1%) with irritable bowel syndrome are embarrassed in school-maladjustment; leaving class early, late coming, a long absence, and a withdrawal.


[The general aspects of the treatment of irritable bowel syndrome]. Irritable bowel syndrome (I.B.S.) is treated with medication, change of life style, various psychotherapies, oriental medicine, and etc., But no specific therapy is recognized useful for all the symptoms derived from I.B.S.. In fact the combination of these therapies are useful. When the therapeutic schedule is planned for individuals, we must keep it in mind that the relief from the symptoms is most important for patients.


[Pharmaceutical treatment of irritable bowel syndrome]. In the treatment of IBS best results could be obtained by implementing a comprehensive program for the patients. This might include a through examination, an explanation of the condition to the patients, psychologic managements, and correction of any bad habits, as well as drug therapy. The aim of drug therapy of IBS is the relief of the symptoms: such as abdominal pain, disturbed bowel function, anxiety or depression. As there is no drug which is effective in relieving the entire range of symptoms, drug should be chosen according to specific symptoms. Tranquilizers and antispasmodics may be the most commonly used drugs, however their efficacy is limited. To postprandial pain antispasmodics or trimebutine are most effective when prescribed before meal. Antidepressant are beneficial for the depressive state. Bulking agents are preferable mainly in relieving constipation, and loperamide is effective in treating diarrhea.


[Synaptic plasticity and gene products]. Synaptic plasticity is thought to be the basic mechanism underlying learning and memory. The cellular mechanisms underlying synaptic plasticity have been extensively investigated in invertebrates and in vertebrates. What is the nature of synaptic plasticity? Can genes and gene products regulate plasticity? If so, how? The behavioral sensitization of the gill-and-siphon-withdrawal reflex of Aplysia is a simple model of plasticity in invertebrates, and can be examined in dissociated cell culture. Using a model of plasticity in cell culture, the molecular cascades of both short-term and long-term sensitization have been investigated, and characterized. Both gene transcription and protein synthesis were shown to contribute to the long-term sensitization. Long-term potentiation (LTP) is a well-characterized model for synaptic plasticity in vertebrates. Many possible cascades have been proposed, but it has not yet been settled whether an increase of transmitter release from presynaptic terminal, an increase of synaptic current, is responsible for the maintenance of LTP. Inhibition of protein synthesis resulted in a failure to maintain LTP over 3-4 hours. Thus, new protein synthesis may be needed for the maintenance of LTP. Induction of so-called immediate early genes that are induced immediately upon depolarization or neurotransmitter stimulation of the neuron has been studied as a possible mechanism underlying LTP. However, there is no good evidence yet implicating gene regulation to be involved in plasticity in vertebrates.


Identification and characterization of the alpha 2D-adrenoceptor subtype in single cells prepared from guinea pig tracheal smooth muscles. Single cells were prepared from guinea pig tracheal smooth muscle and used in binding studies of [3H]p-aminoclonidine. Specific binding of [3H]p-aminoclonidine was saturable to a single class of receptors, with an equilibrium dissociation constant (KD) of 1.62 +/- 0.17 nM and a density (Bmax) of 6.20 +/- 0.78 x 10(4) sites/cell. Competition studies were carried out with several adrenergic antagonists. The rank order of potency was idazoxan = phentolamine > yohimbine > prazosin = corynanthine. These results suggest that the alpha 2-adrenoceptor in guinea pig tracheal smooth muscle represents a single pharmacological subtype, which is designated as alpha 2D.


[Changes in the renin-angiotensin system and the effectiveness of the treatment of hypertension with calcium antagonists, beta-adrenoblockers and diuretics]. Renin response to nifedipine, verapamil, propranolol and hypothiazide was studied in 133 patients with Stage II hypertensive disease. The common mechanism of action in all these drugs was an increase in baseline plasma renin activity (PRA). In patients with a baseline high RPA, propranolol and nifedipine lowered it, whereas verapamil and hypothiazide unchanged it. Group analysis indicated that enhanced PRA during therapy was unassociated with diminished antihypertensive effects of an agent as compared with patients in whom RPA showed no increase. Propranolol was more effective in patients with high PRA than the other agents. The best nifedipine tolerance was recorded in patients with lower PRA.


A hyperpolarizing response induced by glutamate in mouse cerebellar Purkinje cells. In the vertebrate nervous system, glutamate (Glu) receptors are generally known to cause depolarizing responses. We report here a novel type of Glu response in Purkinje neurons of mouse cerebellar slices, namely glutamate-induced hyperpolarization (GH). This response is not due to activation of inhibitory interneurons, because application of tetrodotoxin (TTX), bicuculline, or strychnine did not abolish GH. In addition, GH persisted in a Ca(2+)-free or a low-Cl- solution, which rules out the involvement of gK(Ca) or GABAA mechanisms. Quisqualate (Quis) and trans-1-amino-1,3-cyclopentanedicarboxylic acid (tACPD), which are potent and selective agonists, respectively, for the metabotropic Glu receptor (mGluR), failed to induce GH. L-2-Amino-4-phosphonobutyric acid (L-AP4) was also ineffective. Simultaneous recording of electrical activity and intracellular Ca2+ concentration ([Ca2+]i) showed that GH was not accompanied by [Ca2+]i changes. Voltage clamp experiments showed that GH is due to reduction of a tonically active conductance with a reversal potential around 0 mV. Two possible mechanisms are suggested for GH: (1) changes in the desensitized steady state of ionotropic Glu receptors, or (2) a novel Glu-mediated mechanism.


Truffle melanogenesis: correlation with reproductive differentiation and ascocarp ripening. The present work uses histochemical techniques to investigate the correlation between reproductive differentiation and age of truffles (Tuber aestivum and Tuber melanosporum) with melanin synthesis. The dopa oxidase and tyrosine hydroxylase activities of tyrosinase have been localized within the ascocarp and melanin localization was performed by the Schmorl's reaction. A true tyrosinase is present in truffles, able to oxidize both l-tyrosine and l-dopa. The tyrosinase activity is on in the young ascocarps (in the peridium, hypothecium, and fertile veins) and off in the ripe ones, thus it appears correlated with the age and differentiation of the sporogenic hyphae that arise from the hypothecium; a similar correlation has been previously described in Neurospora crassa, which is an ascomycete as well as the truffles.


Intrinsic pigment-cell stimulating activity in the catfish integument. In keeping with the concept that local factors in the vertebrate integument affect the expression of pigment cells, the present study was directed toward demonstrating the existence of such factors in the skin of the channel catfish, Ictalurus punctatus. This species has a dark dorsal surface in marked contrast to an almost white midventral surface. Pieces of skin from these two surfaces were used to condition culture media, which were in turn bioassayed using the Xenopus neural tube explant system (Fukuzawa and Ide, 1988, Dev. Biol. 129:25). A certain number of neural crest cells grow out from the explant, and many of these are melanized in a culture medium of Steinberg's basic salt solution (BSS). When the BSS was conditioned with either dorsal or ventral skin, a profound increase in both the number of crest cells emigrated from the neural tubes and the percentage of melanized cells was observed. The effects of dorsal skin were stronger than those of ventral skin and were evident on a dose/response basis. Initial fractionation of conditioned BSS with DEAE ion exchange chromatography produced fractions of particular potency in the stimulation of melanogenesis. A similarly conditioned medium based upon Leibovitz's L-15 was used in the primary culture of mature chromatophores, namely, melanophores, iridophores, and xanthophores from tadpoles of Rana pipiens. Both dorsal and ventral conditioned media stimulated iridophores and xanthophores, but seemed to have little or no effect on tadpole melanophores. A melanization inhibiting factor (MIF) from the ventral surface of adult frogs has been suggested as the basis for the light colored ventrum of amphibians, and although the present experiments were not designed to study catfish MIF, the possible existence of such a factor in this species was supported by the results. The total results of this investigation are discussed in the light of the possible presence of a melanization inhibiting factor (MIF) of greater prevalence in the ventrum and a melanization stimulatory factor (MSF) of greater prevalence in the dorsal integument. It is suggested that the light-colored ventral surface of the catfish and other poikilotherms may result from the presence of higher levels of MIF than MSF. Thus, the expression of melanophores is inhibited while that of iridophores is enhanced. In contrast, higher levels of MSF over MIF in the dark dorsal surface would result in melanophore stimulation and inhibition of iridophore expression.


The effect of CGP 37849 and CGP 39551, competitive NMDA receptor antagonists, in the forced swimming test. The present study examined the effects of CGP 37849 and CGP 39551, competitive NMDA receptor antagonists, in the forced swimming test in rats and mice. Administered in a single dose or three times both examined compounds reduced the immobility time in rats. Active doses used in that test either did not change the locomotor activity or decreased it. A similar effect in both tests was shown by active (R)-enantiomers CGP 40116 and CGP 43487. Reduction of the immobility time induced by CGP 37849 and CGP 39551 in the forced swimming test in rats was antagonized by haloperidol and (+/-)-sulpiride, but not by SCH 23390 or prazosin. CGP 37849, but not CGP 39551, also reduced the immobility time in the forced swimming test in mice. The results obtained indicate that CGP 37849 and CGP 39551 induce an antidepressant-like effect in the forced swimming test, probably via an indirect dopamine activation resulting from blockade of NMDA receptors.


Hormonal and neurochemical correlates of various forms of animal "aggression". The majority of studies attempting to evaluate the roles of hormones and neurochemicals in "aggression" concern laboratory rodents, notably rats and mice, with fewer investigations on infrahuman primates. Studies suggest that situations used to assess aggression (e.g., social conflict tests, parental attack, predatory behavior, use of unavoidable electroshock) actually tap a diverse range of motivations whose functions include offense, defense and predation. It is also apparent that ethoexperimental techniques, i.e., those applying ethological methodologies and concepts to laboratory situations, have advantages in assessing the direct and indirect consequences of chemical treatments. In this review, the impacts of hormonal manipulation (by surgery and/or application) and varying neurotransmitters (studied in terms of regional changes and as consequences of drug treatments) on a variety of forms of behavior are assessed. Different tests do show varying responses to common treatments, confirming the heterogeneity of the available paradigms. A brief discussion is provided of which tests are likely to prove most relevant to clinical studies.


Abnormal growth hormone and cortisol, but not thyroid-stimulating hormone, responses to an intravenous glucose tolerance test in normal-weight, bulimic women. Abnormal growth hormone (GH) and adrenocorticotropic hormone (ACTH)/cortisol secretory patterns in response to a glucose load have been observed in underweight anorectic women. The present study was performed in an attempt to establish whether changes in the hypothalamic/pituitary sensitivity to hyperglycemia occur in bulimia in the absence of weight disturbance. Therefore, serum GH, plasma cortisol, and plasma insulin concentrations were measured in eight women with normal weight bulimia and in eight normal women during an intravenous glucose (0.33 g/kg as an IV bolus) tolerance test (IGTT). In addition, since abnormal pituitary hormone responses to a glucose load might reflect alterations in somatostatin (SRIH) release, TSH secretion also was measured, in view of its sensitivity to SRIH inhibition. Both GH and cortisol levels progressively and significantly declined during IGTT in the normal subjects. In the bulimic women, cortisol levels remained unchanged, whereas GH concentrations rose significantly after glucose injection. Plasma cortisol and serum GH levels were significantly higher in the bulimic than in the control subjects. No significant differences between groups were observed in hyperglycemia-induced insulin increments or in TSH decrements. These data indicate that an altered sensitivity to hyperglycemia affects the hypothalamic/pituitary centers controlling the secretion of the counterregulatory hormones GH and ACTH/cortisol in bulimia nervosa. The lack of a simultaneous change in the TSH secretory pattern argues against a possible involvement of SRIH in the pathophysiology of this disorder.


The effect of imipramine after single and repeated administration on the apomorphine response in the acoustic startle reflex in rats. The paper presents the results of studies into the effects of single and repeated (2 or 10 mg/kg po, twice daily for 14 days) administration of imipramine (IMI) on the behavior of male Wistar rats in an acoustic startle response test. Statistically significant attenuation of the animal reactivity was observed after 8 or 14 days administration of IMI in a dose of 10 mg/kg. Apomorphine APO (1.0 mg/kg, ip) enhanced the animals reactivity after repeated (but not single) administration of IMI in a dose of 2 mg/kg. The obtained results confirm the hypothesis about an increase of the dopaminergic system reactivity under the influence of repeated administration of tricyclic antidepressants.


Early response gene induction in astrocytes as a mechanism for encoding and integrating neuronal signals. Astrocytes in vitro readily respond to a wide variety of neuronal and systemic signals by inducing a complex pattern of early response genes (ERGs). The level of complexity is evident in both the ligand-associated expression kinetics and levels of message accumulation as well as in the heterogeneity of response within a population of astrocytes. Ligand-restricted expression of ERG mRNAs suggest that all astrocytes in culture are not alike. Although the ability of glial cells to express ERGs appears to be highly restricted in vivo, one important exception is the category of glial response to injury. Long-term expression of multiple ERG proteins in glial cells stimulated by neuropathological conditions may play an important role in the outcome of brain injury and neurodegenerative disease. The extensive and staggered expression of ERG proteins acting as transcription factors may represent a mechanism for temporally coordinating the genomic program of large sets of genes associated with glial cell response to neuronal dysfunction. As part of the neuronal-glial interdependency, glial ERG expression may encode and integrate the environmental signals associated with neuronal damage and promote the proper restoration of neuronal function. For example, ligand-induced ERG expression regulates the transcription of the nerve growth factor (NGF) gene in glia which could have important functional consequences on neuronal survival and process outgrowth. Future studies will clarify the mechanisms that lead to the expression of ERGs and the subsequent complex, temporally ordered combinatorial consequence of numerous ERG proteins acting as transcription factors impinging upon target gene promoters. Such studies will enrich our understanding of astrocyte-neuronal interaction, clarify how distinct sets of genes in glial cells contribute to the problem and/or solution of neuropathological situations and guide our efforts to diagnose and treat neuropathological conditions.


Partial purification of a receptor for the adipokinetic hormones from Musca autumnalis face flies. Partial purification of the receptors for the neurohormones, diptera corpora cardiaca factors 1 and 2 (DCC1 and DCC2) was achieved. Receptor proteins were obtained from the abdomens of face fly, Musca autumnalis De Geer. Purification methods included detergent solubilization, affinity chromatography, and polyacrylamide gel electrophoresis. Analysis by gel electrophoresis has identified two proteins from this partial purification with relative molecular weights of 45 and 90 kD. A crude receptor preparation was used to develop a ligand binding assay with radiolabeled (tritiated and iodinated) DCC1. Ligand binding was inhibited by 90% when excess unlabeled DCC1 was added to the assay mixture. Ligand binding was optimum at pH 7.5. Binding saturation occurred at approximately 12 picomole radiolabeled ligand concentration. Because DCC1 and DCC2 have been shown to effect the lipid and trehalose levels in the insect an understanding of the neuropeptide-receptor interaction is important for the development of new methods of control of dairy and poultry muscoid flies.


[Therapeutic use of somatostatin analogues in endocrinology]. The recent availability of the long-acting somatostatin analogue, octreotide, has allowed its therapeutical use in a wide variety of human diseases, including some digestive, neoplastic and autoimmune disorders. This review focuses on the treatment of some endocrine disorders with octreotide. Evidence is accumulating that octreotide treatment is effective in improving the cure rate of pituitary surgery in acromegaly by shrinking the tumour size, and in lowering GH and IGF-I levels in the vaste majority of patients. Octreotide is also effective in ameliorating TSH-induced hyperthyroidism in patients with TSH-secreting adenomas. Moreover, octreotide has proved useful in the management of endocrine tumours of the gastroenteropancreatic tract (vipomas, glucagonomas, gastrinomas, insulinomas, and carcinoids) by reducing hormone levels and in some instances the size of the primary and/or metastatic lesions. Besides the above well-established indications there are some other potential indications (non-secreting pituitary tumours, medullary thyroid carcinoma, ectopic Cushing's syndrome, diabete mellitus, Graves' ophthalmopathy, tall children and polycystic ovary syndrome) that still await further investigation. Side-effects of octreotide, particularly the formation of gallstones, should be carefully monitored.


Respiratory burst of neutrophils enhances in hemorrhagic fever with renal syndrome (HFRS). Respiratory burst of neutrophils was studied in 40 serologically verified patients with hemorrhagic fever with renal syndrome (HFRS) using luminol-dependent chemiluminescence (CL). Markedly increased CL has been demonstrated since the first days of the illness in all investigated patients. The level of CL was much enhanced during initial, oliguric, polyuric stages and convalescence. In a case of HFRS complicated by shock CL was approximately 100-fold higher than in healthy controls. It is suggested that neutrophils have an important role in the pathogenesis of HFRS.


Propionate metabolism in cultured human cells after overexpression of recombinant methylmalonyl CoA mutase: implications for somatic gene therapy. Strategies for somatic gene therapy must consider the metabolic consequences of expressing the recombinant gene product in addition to methods for gene transfer and expression. We describe studies of propionate metabolism in cultured cells transfected with methylmalonyl CoA mutase (MCM), the enzyme deficient in mut methylmalonic acidemia. Transfection of MCM into mut fibroblasts restores propionate metabolism to normal levels in a dose-dependent manner. Overexpression of MCM, or the addition of excess propionate, carnitine, or cobalamin, does not increase propionate metabolism in normal human fibroblasts, lymphoblasts, or hepatoma cells, although hepatic cells exhibit > 10-fold higher levels of propionate metabolism. Significantly, the restoration of propionate metabolism in mut fibroblasts is disproportionately greater than the efficiency of transfection, suggesting the presence of a cooperative phenomenon between cells. Intercellular participation in propionate metabolism is evident in cocultures of MCM-deficient and propionyl CoA carboxylase-deficient cells. We conclude that the liver is the preferred target for gene therapy of MCM deficiency because of its greater capacity for propionate metabolism and that cooperation between cells could enhance the biological effect of a subpopulation of cells transformed with recombinant MCM.


Correction of methylmalonyl-CoA mutase deficiency in Mut0 fibroblasts and constitution of gene expression in primary human hepatocytes by retroviral-mediated gene transfer. Methylmalonic acidemia is an often fatal inborn error of organic acid metabolism due to deficiency of methylmalonyl-CoA mutase. The cloning of genes encoding this enzyme and the advent of technologies for gene transfer have introduced the possibility of somatic gene therapy for this disorder. Gene therapy may require replacement of the defective enzyme in hepatocytes, which have a greater capacity for propionate metabolism than other somatic cells and represent the principle physiological site of propionate metabolism. We describe construction of an amphotropic retroviral vector containing the human methylmalonyl-CoA mutase cDNA. This vector is shown to transduce primary MCM-deficient fibroblasts and restore levels of [14C]propionate metabolism by cultures of nonselected cells to normal. This vector will transduce primary human hepatocytes and direct transcription of recombinant human MCM from the integrated provirus. This work demonstrates the feasibility of retroviral-mediated gene transfer of methylmalonyl-CoA mutase into primary human cells, including hepatocytes which represent a difficult, but potentially necessary, target for gene therapy of methylmalonic acidemia.


[Treatments of multiple myeloma in 1992]. In 1992, the Melphalan-Prednisone (M. P.) protocol remains a standard treatment of multiple myeloma even if a lot of new ways have been investigated during the last years. Polychemotherapies may appear better than M.P. for high tumor mass myeloma. Interferon is useful as maintenance treatment after chemotherapy. Combinations of interferon and chemotherapy, during the induction phase, are under evaluation. Because of their toxicity, heavier treatments, with stem cells reinfusion, are being developed mainly with younger patients. Thanks to these approaches the response's rate is increasing but any improvement of survival is still to be demonstrated. Other recent investigations have concerned diphosphonates and immunoregulators. Larger use of these new treatments requires more informations about prognostic factors and their integration in therapeutic strategy of myeloma.


Identification of kinetochores and DNA synthesis in micronuclei induced by mitomycin C and colchicine in Chinese hamster ovary cells. The presence of kinetochore and DNA synthesis in micronuclei (MN) induced in Chinese hamster ovary (CHO) cells by clastogenic and aneuploidogenic substances such as mitomycin C (MMC) and colchicine was determined by immunofluorescence technique using CREST antikinetochore antibodies and anti-bromodeoxyuridine (BrdUrd) antibodies. A cytofluorimetric analysis was also performed. Colchicine significantly increased micronucleated cells at least up to 96 h from the end of treatment. As expected, among colchicine-induced micronucleated cells the majority contained at least one CREST + MN. MMC induced a significant increase in micronucleated cells up to 120 h from the end of treatment and the great majority of MN lacked kinetochore fluorescence, indicating that MMC-induced MN were derived from acentric fragments. However, colchicine and MMC at 48 and 72 h from the end of treatment, induced a significant increase of CREST- and CREST + MN, respectively, suggesting an induction of clastogenicity by colchicine and aneuploidy by MMC. The clastogenic effect of colchicine after 48 h was also confirmed by the presence of chromatid fragments in metaphase cells. A cytofluorimetric analysis indicated that, as expected, colchicine and MMC interfere with the G2/M and S phases, respectively; however, a slight interference of colchicine with the S phase was also observed. DNA synthesis was present in MN and it was in most cases synchronous with synthesis in the main nucleus. The frequency of cells with MN in S phase observed in untreated or MMC-treated cells is in agreement with the proportion of cells without MN showing DNA synthesis. On the contrary, the frequency of cells with MN in S phase observed in colchicine-treated cells was significantly lower than that observed in control and MMC-treated cells.


Tumor formation and sister chromatid exchange induction by ethyl carbamate: relationships among non-pregnant murine females, gravid dams, and transplacentally exposed offspring. Sister-chromatid exchange (SCE) induction and cell cycle kinetics alterations by ethyl carbamate in bone marrow of non-gravid murine Swiss Webster, ICR/Jcl, and C57Bl/6J dams were evaluated, and data from non-gravid females were compared with those previously reported for pregnant dams of the same strains. In addition, lung adenoma induction by ethyl carbamate in gravid Swiss Webster dams, their offspring, and in non-pregnant Swiss Webster females was also determined. Relative cytogenetic and tumor responses in non-gravid and gravid Swiss Webster females and their offspring were compared. In contrast to the increased sensitivity reported for gravid Swiss Webster dams versus ICR/Jcl and C57Bl/6J dams, SCE responses to 1.1, 2.2, or 3.3 mmol/kg of ethyl carbamate in non-gravid females were approximately equivalent among strains. In Swiss Webster and C57Bl/6J (but not ICR/Jcl) strains, SCE responses in non-gravid females at 2.2 and 3.3 were significantly lower than those of their pregnant counterparts. Tumor induction by 3.3 mmol/kg ethyl carbamate paralleled relative SCE induction with Swiss Webster dams, demonstrating a 5-fold increase in the number of tumors relative to their offspring and a 4-fold enhancement of tumor induction relative to their non-pregnant counterparts.


Effects of various bile acids and their sodium salts on development of pepsinogen-altered pyloric glands in rats. Effects of dietary bile acids and their sodium salts on the development of pepsinogen-altered pyloric glands (PAPG) were examined in male WKY/N Crj rats initially given a single dose of 160 mg/kg body weight of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) by gastric intubation. From week 3 the animals were administered basal diet containing 0.5% supplements of cholic acid (CA), deoxycholic acid (DCA), chenodeoxycholic acid (CDCA) or their sodium salts (Na-C, Na-DC and Na-CDC), or 5% ascorbic acid (ASA) or its salt (Na-AS) for 18 weeks. The concentration of DCA and Na-DC was reduced to 0.3% from week 12. At week 20, animals were killed and the numbers of immunohistochemically-demonstrated PAPG were determined. Values were significantly higher with Na-C and Na-CDC than with the corresponding parent acids, and in the Na-C case PAPG development was greater than with MNNG alone. In addition, Na-CDC itself induced the numbers of PAPG significantly. These results suggest that bile salts are possible intrinsic promoters of gastric carcinogenesis. They were without effect, however, on forestomach lesions.


The modulatory effects of tryptamine and tyramine on the S9-mediated mutagenesis of IQ and MeIQ in Salmonella strain TA98. The S9-mediated mutagenesis of IQ and MeIQ in Salmonella strain TA98 was modulated by introduction to the assay of tryptamine or tyramine. Both biogenic amines inhibited or enhanced the mutagenic response as a function of amine concentration, strain of rat used as the S9 source, and the IQ-type mutagen tested. Enhancement of IQ mutagenesis by tryptamine (10-80 microM) was observed in the presence of S9 preparations derived from Aroclor 1254-pretreated Fischer rats; the enhancing effect ceased at tryptamine concentrations > 160 microM. When Sprague-Dawley-S9 or Wistar-S9 were used for activation, the enhancement of IQ mutagenesis by tryptamine shifted to inhibition at tryptamine concentrations > 40 microM, with Sprague-Dawley-S9, and > 20 microM, with Wistar-S9. By contrast, MeIQ-mutagenesis was enhanced by tryptamine (10-160 microM), regardless of the rat strain used as S9 source. Tyramine was a weaker enhancer of MeIQ mutagenesis than was tryptamine and, unlike tryptamine, its inhibitory effects on IQ mutagenesis were observed only with Wistar-S9. Tryptamine (10-80 microM) inhibited cytochromes P450IA1 and P450IA2 activities, monitored by the O-deethylation of ethoxyresorufin and Glu-P-1 mutagenesis in TA98, respectively. These data suggest that the effects of biogenic amines on IQ and MeIQ bioactivation are complex. Furthermore, this study demonstrates that tryptamine and tyramine act both as enhancers (comutagens) and as inhibitors (antimutagens) of IQ and MeIQ mutagenesis, depending on the testing conditions.


The influence of low protein diet on the testicular toxicity of di(2-ethylhexyl)phthalate. Oral administration of di(2-ethylhexyl)phthalate (DEHP) at 1000 mg/kg body weight to adult male albino rats maintained on low protein (LP) diet for 15 d resulted in a greater decrease in absolute and relative weights of the testis and in epididymal sperm count than in those rats maintained on a normal protein (NP) diet. A marked increase in the activity of testicular beta-glucuronidase and gamma-glutamyl transpeptidase (GGT) in the LP-fed animals suggested that LP diet enhanced the vulnerability of Sertoli cells towards DEHP. A greater decrease in the activity of testicular acid phosphatase, lactate dehydrogenase isoenzyme-X (LDH-X) and sorbitol dehydrogenase (SDH) in the LP-fed animals occurred in comparison to NP-fed animals. Degeneration of mature germinal cells in the LP-fed animals on exposure to DEHP suggested that LP diets enhance the susceptibility of the testis towards DEHP.


The relationship between anti-human eye muscle antibodies and thyroid function, anti-TSH receptor antibodies and eye symptoms in Graves' ophthalmopathy. Circulating IgG, IgA, and IgM antibodies to human eye muscle cytosol antigens were studied in 60 patients with Graves' ophthalmopathy using the indirect ELISA method. There was a significant difference in the levels of both IgG and IgA antibodies between the patients with Graves' ophthalmopathy and a control group (p < 0.001). IgA antibodies to eye muscle cytosol antigens were raised in 20 out of 29 patients with proptosis (class 3 ophthalmopathy), in comparison with 31 patients out of the total group of 60 with Graves' ophthalmopathy (p < 0.02). Anti-TSH receptor antibodies (TRAK) were not present in over half of the 31 patients with raised IgA antibodies to eye muscle antigens. However, a significant difference was found between the levels of IgG and IgA antibodies in the TRAK-negative patients (p < 0.05). These findings suggests that both IgG and IgA antibodies to eye muscle antigens might be important in the development of ophthalmopathy.


The prevalence of arboviral, rickettsial, and Hantaan-like viral antibody among schoolchildren in the Nile river delta of Egypt. A serosurvey was conducted during October and November 1989 to estimate the prevalence of selected arboviral, rickettsial, and Hantaan viral antibody among a sample of schoolchildren from 4 villages in the Bilbeis area of the Nile river delta, Egypt. Blood specimens were obtained from subjects aged 8 to 14 years. Enzyme immunoassay testing of the sera indicated that the prevalence of antibody was 9% (21/223) for Sicilian sandfly fever, 4% (8/223) for Rift Valley fever, 3% (15/437) for West Nile virus and 9% (28/315) for Hantaan (HTN) virus. Antibody was demonstrated among 22% (93/418) of the same study subjects against Coxiella burnetti, 53% (199/373) against Rickettsia typhi, and 37% (137/371) against R. conorii.


Light and electron microscopic immunocytochemical localization of glutamine synthetase in the superficial pineal gland of the rat. Glutamine synthetase (L-glutamate:ammonia ligase; EC 6.3.1.2), an enzyme catalysing the ATP-dependent formation of glutamine from glutamate and ammonia, was detected immunocytochemically only in glial (interstitial) cells of the superficial pineal gland of the rat. The results show the important role of pineal glial cells in the metabolism of the presumptive neurotransmitters, glutamate and gamma-aminobutyric acid (GABA) as well as in detoxification of ammonia.


Radioimmunoassay for a new phenothiazine derivative and its application. Fluphenazine-4-chlorophenoxy-isobutyrate ester, a new phenothiazine derivative was synthesized in the Institute for Drug Research Budapest. Radioimmunoassay was developed for the therapeutic monitoring of the drug level after intramuscular depot injection. The fluphenazine hapten was coupled to BSA by mixed-anhydride method. Antisera were produced to this conjugation in New-Zealand white rabbits and were tested for the antibody-titer. The specificity was tested by the cross-reaction with phenothiazine-analogues and other psychotropics. Strong cross-reaction was found with compounds possessing piperazine in side chain (trifluoperazine, perphenazine), but other psychotropic drugs did not react. Tritium-labelled trifluoperazine (spec. activity: 3.5 TBq/mmol) was used as a tracer in the radioimmunoassay. The detection limit was 75 pg with a CV of < 5% in 50 microliters plasma sample (equivalent to 1.5 ng/ml concentration) and a standard curve in the 3 ng/ml-50 ng/ml GYKI-22441 concentration range showed a CV of < 10%. Preliminary pharmacokinetic study was performed in Beagle dogs after intramuscular depot injection with GYKI-22441 in sesame oil in a dose of 0.1 mg/kg. The GYKI-22441 concentration of the plasma samples were measured by the RIA method during a 28-day interval after the treatment and was evaluated by the MultiCalc Immunoassay Data Management program (Pharmacia).


Development of phenylethanolamine N-methyltransferase (PNMT) in cultures of dissociated embryonic rat medulla oblongata. The adrenergic phenotypic marker, phenylethanolamine N-methyltransferase (PNMT) is expressed in a subgroup of catecholaminergic neurons in the brain, as well as in the chromaffin cells of the adrenal medulla. Although PNMT in the rat adrenal is regulated by glucocorticoids, PNMT in the rat brainstem appears not to be regulated by glucocorticoids. Furthermore, little is known about factors required for the differentiation of this specific class of central neuron. The identification of such factors has been hampered not only by the heterogeneity of cell types in the brainstem, of which only a smaller number express PNMT, but also by the lack of a well characterized in vitro system in which the development of these neurons can be studied under defined conditions. The present study addresses this issue by establishing and characterizing a culture system for the study of adrenergic neurons. Dissociated cultures were prepared from embryonic rat medulla oblongata and the expression and development of PNMT was studied using immunocytochemistry and radioisotopic assay of PNMT activity. The survival of PNMT-immunoreactive (IR) neurons in vitro was found to be critically dependent on embryonic age. Numerous PNMT-IR neurons were observed in cultures prepared only from embryos of 46-51 somites (embryonic day E13-13.5). In contrast, cultures containing numerous neurons immunoreactive for tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis, could be successfully established from medulla oblongata of any age between E13 and E16.(ABSTRACT TRUNCATED AT 250 WORDS)


Neuron-enriched cultures derived from spinal cord of 10-day-old chick embryos: influence of neuropeptides on neuronal survival, proliferation and cholinergic expression. The developmental regulation of cell proliferation, survival and cholinergic expression by growth hormone-releasing hormone (GHRH) and somatostatin (SRIF) was investigated in neuron-enriched cultures derived from 10-day-old embryonic chick spinal cord. In this study, 3H-thymidine in corporation into DNA was assessed, using two different applications, in order to determine both cellular proliferation and survival. The rate of neuroblast proliferation in both control and neuropeptide-treated cultures increased or remained the same up to day 6. However, in neuropeptide-treated cultures the magnitude of cell proliferation remained at levels higher than those observed in controls through day 6 and was most significant in SRIF-treated cultures at C4. In all groups, proliferation markedly declined by day 8. Survival of neuronal cells labelled at C4 remained high up to day 12 in all three groups, then drastically declined by day 17. Neuronal survival in the neuropeptide-treated cultures was also higher than in controls. Cholinergic expression, as assessed by activity of choline acetyltransferase (ChAT), responded differentially to neuropeptide treatment. Cultures treated with GHRH (100 nM) exhibited a long term significant enhancement in ChAT activity throughout the culture period, whereas those treated with SRIF (50 nM) expressed a transient decline in ChAT activity. Videometric analysis showed that both neuropeptides enhanced neuronal aggregation, neuritic arborization and neuritic length. These findings lead us to suggest that GHRH and SRIF may provide neurotrophic signals important not only for neuronal proliferation and survival but also for cholinergic neuronal expression. Furthermore, we propose that GHRH possesses specific cholinotrophic properties, whereas SRIF may act as a general neurotrophic factor.


[Currents aspects of H2 receptor antagonists in the treatment of ulcers]. Inhibition of H2 receptors has been the first fully evaluated treatment of peptic ulcer and remains the most widely used mode of therapy. In this review, we summarize the current data on clinical pharmacology, therapeutic indications and results of the four currently used drugs: cimetidine - ranitidine - pepticidine - nizatidine. Their similarities are stressed. Recent data are underlined. The superiority or necessity of a single evening dose is questioned, as well as the clinical importance of tolerance and rebound. The effect on gastric alcohol dehydrogenase is mentioned pending further work on the clinical importance of this discovery. In the acute treatment, the antisecretory potency is of major importance in duodenal ulcer, the duration of treatment in gastric ulcer. Maintenance treatment prevents complication as well as recurrence. H2 receptor antagonists remain a primary treatment of peptic ulcer.


[Therapeutic efficacy of 5-ASA molecules in idiopathic intestinal inflammatory diseases: critical review]. Sulfasalazine has been the most widely prescribed drug for patients with inflammatory bowel disease. Clinical trials have established its usefulness in treating patients with active ulcerative colitis and Crohn colitis and its important role in maintaining remissions in patients with ulcerative colitis. Despite its widespread acceptance, the usefulness of sulfasalazine has been limited by the occurrence of adverse reactions in about 10 to 20% of the patients. Now the aminosalicylates are emerging as a treatment for both ulcerative colitis and Crohn disease. We have critically reviewed the clinical trials assessing the efficacy of 5-ASA molecules. Therapeutic efficacy of 5-ASA appears to be as good as sulfasalazine but causing less adverse effects. In mild to moderate ulcerative colitis relapse, 2g 5-ASA is active while 1 g 5-ASA seems equivalent to 2g sulfasalazine for maintaining remission. 5-ASA enema in the treatment of distal ulcerative colitis is helpful and can replace topical cortisone administration. Administration of 1g 5-ASA enema a day seems to be the best regimen. In case of Crohn's disease, preliminary studies are encouraging but more date are required to define the indications as well as the regimen.


Montenegro skin tests for American cutaneous leishmaniasis carried out on school children in Rio de Janeiro, Brazil: an indicator of transmission risk. Montenegro skin tests were carried out in 1985 and 1987 on two groups of school children in the city of Rio de Janeiro. Group A consisted of 449 children residing in the Jacarepagua district, in areas where transmission of human and canine American cutaneous leishmaniasis (ACL) has been high; this group was considered to be the one at greatest risk of acquiring the infection through L. braziliensis. Group B consisted of 282 children from Bonsucesso, a suburb of Rio de Janeiro that is at a considerable distance from any area where ACL is endemic; this was a lowest risk group, and was thus used as a control. Analysis of the cutaneous test results showed that in Group A, 8.9% of the sample tested positive, whereas in Group B the result was only 2.1%. In group A there were no significant differences in the proportion of positives when analysed according to sex, age and the year when the tests were carried out. A study of the effects associated with place of residence together with other variables was carried out using log-linear regression analysis. It showed that effects arising from place of residence were maintained; that this was the only significant effect; and that it was independent of other variables. The testing of school children using the Montenegro intradermal test was shown to be a useful procedure in the characterization of localities in which there is a risk of ACL infection.


The efficacy of ethion EC (1010 g/l) against cattle ticks in Morogoro, Tanzania. Ethion, an organophosphorus insecticide/acaricide, was found to be effective against the various cattle ticks present in Morogoro Tanzania after it was sprayed on cattle twice weekly for 1 year. The pre- and post-replenishment concentrations of ethion ranged between 0.04 and 0.05%. Ethion was found to be stable in the spraywash. The good efficacy makes ethion a good alternative to dioxathion in tick control in Tanzania.


Fibrinolytic activity in women on oral contraceptive pills: variation due to haemoglobin genotype. Plasma fibrinogen levels and euglobulin lysis time (ELT) were determined in 84 women with haemoglobin genotype AA (HbAA) and in 38 with haemoglobin genotype AS (HbAS), aged 17-35 years, who were on oral contraceptive pills (OCP). The control group included 23 HbAA and 27 HbAS age-matched, apparently healthy women, who had regular menstruation and no history of OCP usage. The controls showed statistically significant elevations in fibrinogen levels in women with genotype HbAS (+13%; P < 0.05) compared with women with genotype HbAA. Among OCP users, the difference in fibrinogen levels (+5%) between HbAS and HbAA women was not statistically significant. The elevation in fibrinogen levels which was restricted to the HbAA women, was probably caused by OCP use, and may be dependent on the Hb genotype. In contrast, the observed elevations in euglobulin lysis time among OCP users (P < 0.005) were independent of Hb genotype. Thus, while OCP may constitute a risk factor for the development of thromboembolism in women, the S-gene may confer partial protection against this development in women who have the HbAS genotype.


Antibody patterns in rabbits showing different levels of susceptibility to an experimental Trypanosoma evansi infection. Using SDS-PAGE and Western blotting, the antibody spectrum of rabbits infected with Trypanosoma evansi to homologous T. evansi antigens was monitored. Animals that developed parasitaemia later or had lower levels of parasitaemia as the infection progressed were considered to have a degree of resistance to the infection. Sera of these resistant animals recognised the T. evansi antigens earlier and subsequently identified more antigens than their susceptible counterparts. The susceptible animals developed patent parasitaemia earlier and had higher parasite counts as the infection progressed, and their sera recognised T. evansi antigens later with fewer parasite components labelled during the course of the infection. These observations demonstrate clear differences between animals in response to T. evansi infections. Selection of T. evansi-tolerant animals on an individual basis may be possible as has been suggested for other trypanosome species.


Non-involvement of nulliparous females in the transmission of bancroftian filariasis. The possible involvement of nulliparous females of Culex quinquefasciatus in the transmission of bancroftian filariasis under field conditions was examined in Pondicherry, South India. Biting nulliparous females that had previously ingested partial blood meals were found infected with microfilariae/L1 stage larvae. None of them harboured infective-stage larvae. These findings suggest that nulliparous females are not involved in the transmission of filariasis. Therefore, their inclusion for estimating the transmission parameters is questionable. However, nulliparous females should also be dissected in order to determine the vector infection rates accurately. Infective larvae were encountered in females of all other age groups.