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PubMed2006

Targeted gene deletion demonstrates that the cell adhesion molecule ICAM-4 is critical for erythroblastic island formation.

Erythroid progenitors differentiate in erythroblastic islands, bone marrow niches composed of erythroblasts surrounding a central macrophage. Evidence suggests that within islands adhesive interactions regulate erythropoiesis and apoptosis. We are exploring whether erythroid intercellular adhesion molecule 4 (ICAM-4), an immunoglobulin superfamily member, participates in island formation. Earlier, we identified alpha(V) integrins as ICAM-4 counterreceptors. Because macrophages express alpha(V), ICAM-4 potentially mediates island attachments. To test this, we generated ICAM-4 knock-out mice and developed quantitative, live cell techniques for harvesting intact islands and for re-forming islands in vitro. We observed a 47% decrease in islands reconstituted from ICAM-4 null marrow compared to wild-type marrow. We also found a striking decrease in islands formed in vivo in knock-out mice. Further, peptides that block ICAM-4/alpha(V) adhesion produced a 53% to 57% decrease in reconstituted islands, strongly suggesting that ICAM-4 binding to macrophage alpha(V) functions in island integrity. Importantly, we documented that alpha(V) integrin is expressed in macrophages isolated from erythroblastic islands. Collectively, these data provide convincing evidence that ICAM-4 is critical in erythroblastic island formation via ICAM-4/alpha(V) adhesion and also demonstrate that the novel experimental strategies we developed will be valuable in exploring molecular mechanisms of erythroblastic island formation and their functional role in regulating erythropoiesis.

Animals↗
PubMed1996

Child neurology: past, present, and future.

The emergence of child neurology as an independent discipline was enhanced by pediatricians developing an organ system approach to the study of illnesses of children and indirectly by their interest in normal psychomotor development, a function of maturation of the nervous system. Technological innovations, advances in neuroimaging, and the development of therapeutic agents directly benefited patients with neurologic disorders and enhanced the role of the neurologist. The American Board of Psychiatry and Neurology was founded in 1934. It formally recognized child neurology as a special area of expertise in 1959 when Sidney Carter was appointed as a director and the clinical examination was changed to include an hour devoted specifically to child neurology. In 1969, special certificates in child neurology were issued. Thirty candidates, or more, have successfully passed the examination each year since then. As of Spring 1995, there had been 1007 certificates issued to child neurologists. In 1991 and 1992, alternative pathways to training in child neurology were developed. These substituted a year of internal medicine or a year of basic neuroscience research for 1 year of pediatrics. Relatively few child neurologists have chosen these alternative pathways. The performance of child neurologists, on Board examinations has generally been comparable to that of adult neurologists with the latter holding a slight edge. In 1990, a study of first-time takers of the certification examination was begun. In child neurology, 54% of the cohort passed both parts I and II on their first attempts, and as of July 1995, 81% of the original cohort was certified. In adult neurology, the comparable rates are 56% and 84%. Accreditation of child neurology training programs began in 1987. As of November 1995, there were 75 accredited child neurology training programs, which offered slightly over 200 total positions. Changes in health care delivery are currently taking place and have placed greater emphasis on primary care physicians. Proposals have been put forth that, if enacted, will reduce the current number of medical students and also reduce the total number of residents being trained. Reductions as great as 40% of current resident number have been proposed. Child neurology will need to reassess our current work force and training needs and re-evaluate our role in the training of primary care physicians. There appears to be a paradox. Explosions of information in both the basic and clinical neurosciences have directly benefited our patients, but yet our continued growth as a subspecialty is being challenged.

Accreditation↗
PubMed1989

Immunochemical analyses of C57BL/6J monoclonal anti-alpha (1----3) dextran antibodies.

Twelve C57BL/6J anti-B-1355S monoclonal antibodies (five IgM lambda and seven IgM kappa) were characterized immunochemically by binding and inhibition ELISA. All 12 were negative for the expression of the cross-reactive idiotype (IdX) of BALB/c mice, as expected from previous work; no kappa IdX+ antibodies have been reported and IdX- lambda class antibodies were observed in B-1355-Con A induced immune sera [Geckeler W., Blomberg B., dePreval D. and Cohn M. (1977) Cold Spring Harb. Symp. quant. Biol. 41, 743-748]. The antibodies studied bind to B-1355 coated plates and this binding is inhibited by B-1355 but not by dextrans B-512 (F) or B-742S; the latter two have no linear alpha (1----3; 1) linkages. The nomenclature of Jeanes is used [Jeanes A. (1986) Molec. Immun. 23, 999-1028]; alpha (1----3; 1) refers to glucosyl diose residues linked alpha (1----3) linearly. In the case of B-1355 these linear stretches alternate with alpha (1, 6) linkages and are non-contiguous; alpha (1----3; b) refers to the linkage at a branching residue, e.g., a 1,3,6 linked moiety. The IgM kappa class antibodies are not inhibited by nigerose or nigerantetraose, suggesting that they have binding site sizes which are unusually large for B-1355 specific antibodies. The five IgM lambda antibodies are inhibited identically by equimolar amounts of nigerose and nigerantetraose, suggesting that their binding sites accommodate a disaccharide epitope. These antibodies are also inhibited by the alpha (1----6), alpha (1----4) triose, panose. The kappa class antibodies do not bind to alpha (1----3)-diglucosyl-(nigerosyl; N)-BSA. Four of the five lambda class antibodies show weak binding to N-BSA, while the fifth binds N-BSA better but less well than MOPC 104E (the BALB/c myeloma protein). All 12 antibodies are unique when compared to BALB/c antibodies derived from B-1355 immunization. The primary response of 15 C57BL/6J mice to B-1355 was re-assessed for kappa and lambda class antibody contribution. A patchy lambda class response was observed suggesting that previous lambda class responses may have been overlooked.

Animals↗
PubMed1989

Discovery of the pre-erythrocytic stages of a saurian malaria parasite, hypnozoites, and a possible mechanism for the maintenance of chronic infections throughout the life of the host.

Re-examination of tissue sections from four Takydromus tachydromoides (Sauria: Lacertidae) naturally infected with Plasmodium sasai found liver parenchymal cells, containing uninucleate parasites which may correspond to the hypnozoite stage of primate malaria parasites, schizonts and segmenters in parenchymal cells, and hepatic macrophages which contained numerous schizonts. Following destaining of the original H&E and prolonged restaining with warm Giemsa stain, encysted schizonts, protected by a hyaline wall, were discovered in the connective tissue or capillary endothelium of lung, liver, brain, heart, pancreas, kidney, intestine wall, testis, and both intra- and intermuscularly in the femoral muscles. Unencysted schizonts in the pulmonary endothelium apparently represent the phanerozoic stages, which, following encystment in the various tissues, are recognized as a new stage in the life cycle of reptilian malarial parasites, the chronozoic schizonts. A hypothesis is presented to describe the life cycle of P. sasai, which may be characteristic of other saurian malaria parasites. It interprets the sequence of pre-erythrocytic stages found as follows: sporozoites enter hepatic parenchymal cells where some may become dormant as hypnozoites, and others form cryptozoic schizonts. The cryptozoites parasitize hepatic macrophages and form metacryptozoic schizonts. Metacryptozoites produce phanerozoic schizonts in the capillary endothelium and connective tissue of the lung and other organs. Phanerozoites and possibly metacryptozoites then invade the erythrocytes to begin the erythrocytic cycle. Some of the phanerozoites in endothelium, connective tissue and skeletal muscle become encysted as chronozoic schizonts, and their progeny, chronozoites, renew the erythrocytic cycle throughout the life of the host and produce seasonal relapses of gametocytemia, in spring, at the end of hibernation by the lizard.

Animals↗
PubMed2000

Mate fidelity and breeding site tenacity in a monogamous sandpiper, the black turnstone.

We examined the relationship between mate fidelity and breeding site tenacity during a 5-year study of the black turnstone, Arenaria melanocephala, a socially monogamous sandpiper breeding in subArctic Alaska. We tested the predictions of several hypotheses regarding the incidence of divorce and the benefits of fidelity to mate and breeding site. Interannual return rates to the breeding grounds (88% for males, 79% for females) were among the highest yet recorded for any scolopacid sandpiper, and 88% of returning birds nested on their previous year's territory. The annual divorce rate was only 11%, and mate fidelity was significantly linked to fidelity to territory but independent of sex and year. Males arrived in spring significantly earlier than their mates and interannual fidelity was influenced by the relative timing of arrival of pair members. Reunited pairs had significantly higher fledging success than new pairs formed after death or divorce. The incidence of divorce was unrelated to reproductive success the previous year, although birds nested significantly further away after failure than after a successful nesting attempt. Sightings of marked individuals suggested that members of pairs do not winter together, and breeding site tenacity provides a mechanism through which pair members can reunite. We reject the 'incompatibility' hypothesis for divorce in turnstones, and our data contradict predictions of the 'better option' hypothesis. Alternatively, we propose the 'bet-hedging' hypothesis to explain the occurrence of divorce, which transpires when an individual pairs with a new mate to avoid the cost of waiting for a previous mate to return. Such costs can include remaining unmated, if the former mate has died, or experiencing lower reproductive success because of delayed breeding. Copyright 2000 The Association for the Study of Animal Behaviour.

Journal Article↗
PubMed2004

Influence of fluoxetine on positive and negative affect in a clinic-based smoking cessation trial.

RATIONALE: Fluoxetine improves affect in clinical syndromes such as depression and premenstrual dysphoric disorder. Little is known about fluoxetine's influence on mood changes after quitting smoking, which often resemble sub-clinical depression. OBJECTIVES: The present study, a re-analysis of previously published data (Niaura et al. 2002), examined fluoxetine's effect on changes in negative and positive affect following quitting smoking. METHODS: Adult smokers (n=175) without clinically significant depression were randomized on a double-blind basis to receive fluoxetine hydrochloride (30 or 60 mg daily) or placebo for 10 weeks in combination with cognitive-behavioral therapy (CBT) for smoking cessation. We postulated that fluoxetine would beneficially influence post-cessation changes in positive and negative affect. RESULTS: Mood change across treatment was analyzed using mixed linear modeling controlling for initial level of nicotine dependence, plasma fluoxetine metabolites, and change in cotinine (a nicotine metabolite) at each visit. Relative to placebo, those on 60 mg fluoxetine experienced an elevation in positive affect that increased across time [t(526)=2.50, P=0.01], and a reduction in negative affect that returned to baseline across time [t(524)=2.26, P=0.02]. There were no differences between 30 mg and placebo on changes in positive or negative affect. CONCLUSIONS: Results indicate that 60 mg of fluoxetine improves both positive and negative mood states after quitting smoking and that diminished positive affect may be an overlooked affective response to smoking cessation.

Adult↗
PubMed2005

Cerebral palsy improvement achieved by coordination dynamics therapy.

Low-intensity coordination dynamics therapy, including crawling, treadmill walking, jumping on spring-board and exercising on a special coordination dynamics therapy device, was applied for 3 months (4 hours therapy per week) to 8 cerebral palsy patients (average age 15 years, range 7-27). All patients improved. The organization of the CNS, quantified by the low-load coordination dynamics between arm and leg turning movements, when exercising on the special coordination dynamics therapy device, improved by 46 +/- 17% (range 33-60%) for forward and by 48 +/- 15% (range 22-66%) for backward moving. Also improved the exercised crawling, jumping and walking, although not as much as the CNS organization quantified by coordination dynamics. The motor programs of the tibialis anterior, gastrocnemius, biceps brachii, and triceps brachii muscles, measured by surface electromyography (sEMG), improved only little. Evidence is provided for sEMG being a very suitable tool for optimizing the movement performance and the therapy since sEMG records show under what exercise conditions the recorded motor programs are best. However coordination dynamics, i.e. the integrative parameter for quantifying CNS organization, is better to show the progress in CNS functioning than movement and EMG improvements. When the patients stopped therapy, the value of their coordination dynamics, worsened 24% after 6 months. In one patient the coordination dynamics therapy was continued intensively for further 3 month, including 20 hours exercise per week. The value of the coordination dynamics even improved altogether by 85% and 82% for forward and backward turning movements respectively, and simultaneously movements, vegetative (sleep) and higher mental functions (aggressivity, learning capacity) showed strong improvements. The improvements of coordination dynamics for low-intensity therapy (46%, for forward movements) and additional high-intensity therapy (85%) lie within the recovery range for stroke (70%) and brain injury (69%) after 3 months of intensive coordination dynamics therapy. There is therefore indication that the CNS functioning in cerebral palsy patients can be improved by learning as much as the CNS functioning can be repaired by re-learning in stroke and brain injury. The improvement of the CNS functioning suggests that cerebral palsy can partly be cured if intensive coordination dynamics therapy is administered for 1 to 2 years. It is further suggested that inabilities, including mental inability, are diseases which can partly be cured rather than inabilities.

Adolescent↗
PubMed1997

Seasonal changes in spermatogenesis and testicular steroidogenesis in the male black bear Ursus americanus.

American black bears, Ursus americanus, are seasonal breeders with a mating season in late spring to early summer. The objectives of this study were to determine whether there are seasonal changes in spermatogenesis and immunolocalization of testicular steroidogenic enzymes, and to correlate these changes with peripheral steroid concentrations. Three captive mature bears were maintained in open cages during the summer season and provided with chambers for denning during the winter. Testicular biopsies and blood samples were obtained from anaesthetized bears on 12 March, 15 June, 12 October and 15 January. Steroidogenic enzymes were immunolocalized using polyclonal antisera raised against bovine adrenal cholesterol side-chain cleavage cytochrome P450 (P450scc), human placental 3 beta-hydroxysteroid dehydrogenase (3 beta HSD), porcine testicular 17 alpha-hydroxylase cytochrome P450 (P450c17) and human placental aromatase cytochrome P450 (P450arom). Spermatogenesis changed seasonally: spermatogonia and degenerating spermatocytes were observed in October; spermatogonia and primary spermatocytes were present in January; spermatogonia, spermatocytes and round spermatids were present in March; and spermatogonia through spermatozoa were present in June. P450scc and P450c17 were immunolocalized in spermatids and Leydig cells in June, whereas in October these enzymes were present only in Leydig cells. 3 beta HSD was localized in Leydig cells in June and October with more intense staining in June. Localization of P450arom changed seasonally: no immunostaining in October; positive immunostaining in Sertoli cells in January; more extensive immunostaining in Sertoli cells, peritubular-myoid cells and round spermatids in March; and strong immunostaining in Sertoli cells and round and elongating spermatids in June. Serum testosterone and oestradiol concentrations changed seasonally: testosterone and oestrogen were low in October and January, slightly higher in March, and high in June. The present study demonstrates that in the black bear seasonal changes in spermatogenesis are accompanied by changes in the immunolocalization of testicular steroidogenic enzymes that are correlated with changes in serum testosterone and oestradiol concentrations. The presence of P450arom in Sertoli cells at the beginning of testicular recrudescence suggests that aromatase and oestrogen may play a role in re-initiating spermatogenesis.

3-Hydroxysteroid Dehydrogenases↗
PubMed2004

Identification of extracellular enzyme producing thermophilic bacilli from Balcova (Agamemnon) geothermal site by ITS rDNA RFLP.

AIMS: Molecular characterization of extracellular enzyme producing thermophilic bacilli from Balcova geothermal site. METHODS AND RESULTS: Three types of geothermal samples were collected: mud, re-injection water, and samples from uncontrolled hydrothermal vents. Isolates grown at 55 degrees C in culture media prepared in sterilized re-injection water, were screened for extracellular enzyme activity by using eight different substrates: casein, carboxymethylcellulose, pectin, polygalacturonic acid (PGA), soluble starch, Tween 20 and 80, and xylan. In total, 109 thermoaerophilic isolates were selected. All of the isolates could hydrolyse Tween 20 (100%) but not Tween 80. Soluble starch was hydrolysed by 96%, casein by 55%, xylan and carboxymethylcellulose by 9%, and pectin and PGA by 2% of the isolates. The isolates were grouped into 14 different homology groups by the restriction pattern analysis of 16S-internal transcribed spacer (ITS) rDNA RFLP. Each of the RFLP groups was also studied by 16S rRNA gene partial sequence analysis. Plasmid DNA profiles revealed that 15 of the isolated strains contained small plasmid DNA molecules ranging in size from 12 000 to 35 000 bp. CONCLUSIONS: Combined analysis of 16S-ITS rDNA RFLP and 16S rRNA gene partial sequence results indicated the presence of novel or existing species of Anoxybacillus (nine species) and Geobacillus (three species). SIGNIFICANCE AND IMPACT OF THE STUDY: In this study 16S-ITS rDNA RFLP was applied for the first time to differentiate thermophilic bacilli. It was also the first study on thermophilic bacilli of Balcova geothermal site.

Bacillus↗
PubMedDate not supplied

[80 years' of internal medicine education at the medical school of the university in Belgrade (1922-2002)].

ORGANISATION OF TEACHING INTERNAL MEDICINE: The Department for Internal Medicine and Internal Clinics were founded in spring 1922. Dr. Radenko Stankovic and Dr. Dimitrije Antic were appointed as part-time Professors, while Dr. Aleksandar Ignjatovski, a former Full-time Professor of the Warsaw University, was appointed as professor under contract. A year later, Dr. Aleksandar Radosavljevic was appointed as Part-time Professor. In the General State Hospital and Military Hospital, certain wards were turned into clinics. II and III Internal Clinics were situated in the barracks, while the Propedeutic and I Internal Clinics were located in the Military Hospital. Upon the construction of the buildings of the Internal Clinic and General State Hospital, the Propedeutic and I Internal Clinics were permanently placed in the new building, and II and III Internal Clinics in the General State Hospital. Teaching of Internal Medicine started 31 October 1922. Dr. R. Stanko vic delivered a lecture in Propedeutics for students of the fifth term. This date marks the beginning of teaching internal medicine at the newly established School of Medicine, University of Belgrade. Dr. A. Ignjatovski started lecturing Internal medicine 23 March 1923, whereas Dr. D. Antic and and Dr. A. Radosavljevic also delivered lectures in the areas of Internal Medicine within their professional scope. At the beginning, the clinics belonged to the General State Hospital. It was impossible to teach successfully in hospital, therefore upon the professors' request, the clinics were separated and thus became the institutions belonging to the School of Medicine-educational institutions, while hospitals were health institutions. The rule was 'one professor--one clinic'. After the Second World War, teaching Internal Medicine was begun in demolished buildings in very difficult financial circumstances. The Propedeutic Internal clinic was renamed IV Internal Clinic, which continued dealing predominantly with cardiology. III and IV Internal Clinics were placed in the building of the Internal Clinic, II Internal Clinic remained in the General State Hospital and I Internal clinic was moved to the new building of the Institute for Oncology. REFORMS OF SCHOOL OF MEDICINE AND ITS CLINICS: Out of the School of Medicine, Veterinary Medicine, Pharmacy and Stomatology, the Grand Medical School was formed. After several years, it was closed down, and the schools were returned to the University. The most important alteration was made when, according to the Law, the Clinics were isolated from the School and proclaimed hospitals, health institutions and organised into the Clinical Centre of the School of Medicine, then University Clinical Centre, and finally the Clinical Centre of Serbia as the teaching base. Within city hospitals, later on Clinical Hospital Centres, the teaching bases for Internal Medicine and other subjects were established, with introducing self-management and the Law on Associated Labour with independent financing, there came hard times for the School of Medicine and Clinics. The Law had negative effects upon the Schools--University and it had to be abolished. The next step in the reform was merging of I and II Internal Clinics into the Internal B Clinic, within the Clinics, out of specialized departments for certain branches of Internal Medicine there were formed clinics (Clinic for Cardiology, Pulmonology, Endocrinology, Diabetes and Metabolic Diseases and so on). The Internal A Clinic was situated in the building of the General State Hospital and the Internal B Clinic in the building of the Internal Clinic, where there were also III and IV Internal Clinics. CONSTRUCTION OF CLINICAL CENTRE: After a long time, there were created conditions for constructing a new building for locating clinics--the Clinical Centre, the foundation stone was laid 20 October 1976, and the first phase including the polyclinics was completed 1 January 1983. The preliminary concept of the bed capacity of 30 to 50 at Internal Clinics was altered and the Institutes were established. Firstly, the Internal Clinics was altered and the Institutes were established. Firstly, the Internal A and B Clinics were joined into the Basic Organisation of Associated Labour Internal Medicine. Afterwards, certain specialized Internal Clinics with respective surgical branches were linked and the Institutes were established (Institute for Cardiovascular Diseases, Institute for Pulmonology and Tuberculosis, Institute for Endocrinology, Diabetes and Metabolic Diseases and so on). The Institutes were located at the former buildings of the clinics as well as the vacant buildings of the Military Medical Academy, which moved into the new modern building at Banjica. Thus, Internal Medicine was organisationally closed down, while assistants and professors were gathered at the Department for Internal Medicine which represented the organisation for teaching Internal Medicine. Professors and assistants from certain specialized Institutes taught certain disciplines, and at the Clinical Hospital Centre bases where teaching staff was deficient, the Clinical Centre professors were of assistance. A colossal Emergency Centre was established within the Clinical Centre, providing services for the whole of the Republic and beyond, which is completely illogical since it should be the Republic Health Institution rather than the part of the Clinical Centre. Also, the Clinical Centre as the health institution is enormous and represents the teaching base. The first directors were lawyers and, as of recently, professors of the School of Medicine. By separating Clinics from the School of Medicine, teaching and allied staff have a double working status as unique, so financially more potent health sector has gradually pushed aside the teaching one being rather underestimated in such organisation. Ministers for Health and Directors of the Clinical Centre great executive power for appointing directors. There started an uprecedented systematic degradation of teaching staff, since assistant professors or newly appointed associated professors were appointed as managers over their own teachers, full-time professors. It is unusual that those young people accepted their big, delicate roles to manage their professors in front of whom they had taken their board exams or defended their masters' and doctoral theses in not so distant past. There has only been one case of a junior professor refusing to be the director of his own professor. The stationary section of the Clinical Centre including the Internal Clinics was completely constructed but has not been equipped or occupied yet. Deans were authoriZed, by the University Law, to appoint professors and assistants. Thus, a large number of them were appointed without any high scientific and teaching quality standards, and afterwards promoted, while certain highly competent professors were dismissed from the University. By altering the system and passing the new University Law, the situation has improved and the University is returning to its roots. Today, such Clinical Centre as the teaching base of the School of Medicine and the School itself represent a big problem, primarily the financial one. Thus, it will be very hard to re-establish the right relationship between these two institutions.

Education, Medical↗