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Neuromotor deficits in children with the 22q11 deletion syndrome.

The 22q11 chromosomal deletion syndrome (22q11DS) is associated with a heterogeneous physical phenotype, neurocognitive deficits, and increased risk of later psychiatric illness. Sporadic clinical reports suggested motor differences, but quantitative studies of movement in children with 22q11DS are rare. If present in a majority of affected school-age children, characterization of neuromotor deficits may prove to be critical for intervention, neurocognitive test interpretation, and understanding etiology. We administered the Movement Assessment Battery for Children to 72 children ages 4.3 to 16.1, including 49 children confirmed positive for the 22q11 deletion and 23 control siblings. We predicted a higher frequency of global and domain impairment in manual dexterity, eye-hand coordination, and balance among affected children. Ninety-four percent of affected children had marked neuromotor deficits, and group scores differed broadly for both global and subarea measures. Secondary analyses showed no impairment differences between younger and older children with 22q11DS, and longitudinal trajectories for 12 affected children suggested stability of deficits over 3-year intervals. Neuromotor deficits in children with 22q11DS occur early in development, continue throughout the school-age years, should be considered in the interpretation of motor-based achievement and IQ tests, and require targeted and ongoing remediation throughout childhood and adolescence. Further studies examining the specificity of motor impairment to 22q11DS are needed.

Adolescent↗

Individual manifestations of Parkinson's disease after ten or more years of levodopa.

Twenty-five consecutive patients with Parkinson's disease, who had been on levodopa for 10 or more years, were studied. Over the 12.9 years of treatment, the average Northwestern Disability Score had increased from 9.6 to 18.9. By this measure, 24 of 25 patients were worse, and one was unchanged. The progression of disability did not involve all pretreatment parkinsonian features equally in any of the patients studied. Instead a distinctive pattern of deterioration was seen: postural reflexes worsened in 24/25, speech in 24/25, and gait in 22/25. In contrast, rigidity was improved or unchanged in 17/25, tremor was improved or unchanged in 17/17, while handwriting was improved or unchanged in 21/22. Finger dexterity which was improved in 5/25, unchanged in 15/25, and worse in 5/25 which seemed to be between these two extremes. These observations demonstrate that some signs of parkinsonism can remain quite responsive to levodopa for more than 10 years while at the same time other manifestations of the disease are no longer as responsive to this form of treatment.

Gait↗

Is focal hand dystonia associated with psychopathology?

The purpose of this study was to determine if patients with focal hand dystonia have any significant psychopathology. We studied 20 patients with hand cramps who were participating in a therapeutic trial of botulinum toxin injections. Patients were interviewed and administered the Minnesota Multiphasic Personality Inventory (MMPI). Beck Depression Inventory, Spielberger State-Trait Anxiety Scale, a finger tapping test, and a choice serial reaction time test. Behavioral ratings were also obtained. Group statistics indicated that all personality scale scores and performances on motor tasks were within normal limits. Four out of 20 patients demonstrated mild depression. Trait anxiety scores were higher than state anxiety scores, suggesting that receiving medical treatment had a beneficial effect on mood. The number of depressive symptoms endorsed on the MMPI was correlated with reaction time speed but not finger dexterity. None of the 20 patients reported a remarkable psychiatric history. These results indicate that hand cramps are not associated with serious psychopathology.

Adult↗

Early-onset Parkinson's disease.

The study was conducted on 120 patients (76 men and 44 women) affected by idiopathic Parkinson's disease (IPD) responsive to L-dopa and observed for many years. Sixty had clinical onset between the ages of 20-40, representing 10.2% of our PD population; in the others the symptoms began after the 40th birthday. The two groups were matched for sex and length of illness. In all patients a diagnosis of IPD depended on history and clinical and neuroradiological findings. Clinical, pharmacological, evolutive, and epidemiological data were collected on all patients. Thirty-six patients from each group performed motor dexterity tests (reaction time to expected and unexpected stimuli) and cognitive tests (Wechsler Adult Intelligence Scale. Benton, Short tale, and Zazzo's speed and accuracy test). To assess the prevalence of dementia and the severity of psychiatric side effects of L-dopa administration, the 60 patients with early-onset PD were compared with 134 consecutive unselected PD patients. Five percent of early-onset PD patients had a family history of the disorder. Our study showed that early-onset PD does not differ fundamentally from the late-onset form except that the former is characterized by a more rapid establishment of the full-blown parkinsonian clinical picture and deterioration of the therapeutic efficacy of L-dopa, with an earlier appearance of side effects. The results of our neuropsychological investigations suggest that early-onset PD may be a "pure" form of extrapyramidal compromise with exclusively motor manifestations.

Adult↗

Effect of adding the D1 agonist CY 208-243 to chronic bromocriptine treatment. I: Evaluation of motor parameters in relation to striatal catecholamine content and dopamine receptors.

A group of four cynomolgus monkeys previously rendered parkinsonian by the toxin 1-methyl-4-phenyl,1,2,3,6-tetrahydropyridine (MPTP) were observed in locomotion cages equipped with photocells during four periods of 7 days during which they received saline or two doses of the D1 agonist CY 208-243. The larger dose of 0.5 mg/kg produced a significant increase in locomotion in three of four animals. A second group of eight monkeys also previously rendered parkinsonian by MPTP and having received no other treatment were given a daily treatment of bromocriptine 1.66 mg/kg orally daily during 4 weeks. In four of the animals, after a week on bromocriptine alone, the D1 agonist CY 208-243 was added in increasing doses of 0.05, 0.1, and 0.5 mg/kg. The motor response as measured by locomotion, hand dexterity, and a disability score improved progressively at least in some of the animals on bromocriptine alone. The addition of CY 208-243 produced a more striking improvement of all three parameters, which appeared to be dose dependent. Biochemical analysis of the brain of these animals plus one control and one MPTP untreated monkey showed a > 90% loss of dopamine in the striatum in six of the eight treated monkeys. Both D2 and D1 dopamine receptors were increased in density by denervation, but both treatments abolished this increase for the D2 receptors while increasing the affinity of the D1 receptors.

3,4-Dihydroxyphenylacetic Acid↗

A multicenter double-blind placebo-controlled trial of pergolide as an adjunct to Sinemet in Parkinson's disease.

Three hundred and seventy-six subjects with advanced Parkinson's disease participated in a prospective, double-blind placebo-controlled study of the dopamine agonist pergolide mesylate as an adjunct to Sinemet. At 6 months, patients randomized to pergolide had a statistically significant improvement in total Parkinson's score, scores of activities of daily living, motor function, number of "off" hours, Hoehn and Yahr stage, and numerous parameters of parkinsonian function including bradykinesia, rigidity, gait, and dexterity. This benefit was obtained with the addition of a mean dose of 2.94 mg of pergolide, which permitted a 24.7% reduction in dose of levodopa. Adverse reactions were, for the most part, mild, reversible, and not of major clinical significance. No significant cardiac or electrocardiographic abnormalities were detected. This study demonstrates that pergolide mesylate, as an adjunct to levodopa, is an effective antiparkinsonian agent that provides clinical improvement while permitting a reduction in levodopa dose.

Activities of Daily Living↗

Microsurgical skills training: a new concept for simulation of vessel-wall suturing.

Initiation of training in basic microsurgical suturing technique usually involves placing sutures on incisions made on latex sheets, and then progressing to a live anastomosis in a rat model. A straight incision resembles a skin incision and does not mechanically simulate vessel wall suturing, and various modifications and innovative prosthetic models continue to be developed to bring the simulation closer to a live anastomosis. We developed two models which closely simulate vessel wall suturing and require the application of similar skills by the trainee. In the first model (the "I" model), the straight 1-cm incision is converted to an I-shaped incision which increases the instability of the edge to be sutured, depending on the length of the horizontal incision, putting greater demand on dexterity in suture placement similar to a vessel. The length (1 cm) of the edge does not constrain the spacing of the sutures and can be used for the beginner. The second model (the "double triangle" model) creates a narrow and unstable edge for placing sutures, and puts greater demand on bimanual coordination and instrument control for training in accuracy of suture placement as well as spacing between sutures. Both these models are cost-effective and simple to construct.

Anastomosis, Surgical↗

Altered regional brain glucose metabolism in Duchenne muscular dystrophy: a pet study.

The basis for cognitive impairment in Duchenne muscular dystrophy (DMD) is not well understood but may be related to abnormal expression of dystrophin in brain. The aim of this study was to determine whether regional brain glucose metabolism is altered in children with DMD and whether such metabolic disturbances are localized to regions shown to be normally rich in dystrophin expression. Ten boys (mean age, 11.8 years) with DMD and 17 normal adults as a control group (mean age, 27.6 years) underwent 2-deoxy-2[(18)F]fluoro-D-glucose positron emission tomography (PET) and neuropsychological evaluation. The PET data were analyzed by statistical parametric mapping (SPM). The SPM analysis showed five clusters of decreased glucose metabolism in children with DMD, including the medial temporal structures and cerebellum bilaterally and the sensorimotor and lateral temporal cortex on the right side. At the voxel level, significant glucose hypometabolism was found in the right postcentral and middle temporal gyri, uncus, and VIIIB cerebellar lobule, as well as in the left hippocampal gyrus and cerebellar lobule. The neuropsychological profile of the DMD group revealed borderline nonverbal intellectual functioning, impaired manual dexterity bilaterally, borderline cognitive functioning, and internalizing behavioral difficulties. Our findings demonstrate region-specific hypometabolism, as well as cognitive and behavioral deficits in DMD children. As the regions showing hypometabolism on PET include those normally rich in dystrophin expression, it will be important to determine whether the hypometabolic regions also show cytoarchitectural abnormalities related to the lack of dystrophin.

Adolescent↗

Zero-inflated models for regression analysis of count data: a study of growth and development.

Poisson regression is widely used in medical studies, and can be extended to negative binomial regression to allow for heterogeneity. When there is an excess number of zero counts, a useful approach is to used a mixture model with a proportion P of subjects not at risk, and a proportion of 1--P at-risk subjects who take on outcome values following a Poisson or negative binomial distribution. Covariate effects can be incorporated into both components of the models. In child assessment, fine motor development is often measured by test items that involve a process of imitation and a process of fine motor exercise. One such developmental milestone is 'building a tower of cubes'. This study analyses the impact of foetal growth and postnatal somatic growth on this milestone, operationalized as the number of cubes and measured around the age of 22 months. It is shown that the two aspects of early growth may have different implications for imitation and fine motor dexterity. The usual approach of recording and analysing the milestone as a binary outcome, such as whether the child can build a tower of three cubes, may leave out important information.

Binomial Distribution↗

Conditions that support long-term production of osteoclast progenitors in vitro.

To better understand the mechanisms of osteoclast precursor development from hematopoietic stem cells, we examined the conditions that support the production of osteoclast progenitors, osteoclast colony-forming units (CFU-O), from long-term bone marrow cultures established under myeloid (Dexter's) and lymphoid (Whitlock and Witte's) conditions. Nonadherent cells harvested weekly from myeloid or lymphoid long-term cultures were assayed for CFU-O-derived colony formation in agar in the presence of a murine osteoclast colony-stimulating factor. The myeloid system supported CFU-O production for weeks, but the system produced many other types of myeloid colonies and cells as well, and quantification of CFU-O-derived colonies was difficult. The lymphoid long-term culture system also produced CFU-O; however, CFU-O production in the lymphoid system appeared more selective than in the myeloid system, but was transient. Interestingly, the addition of medium containing G-CSF to these cultures greatly enhanced (> 200%) the CFU-O production. This enhanced CFU-O production was confirmed using bone marrow cultures established on a defined marrow stromal cell line under lymphoid conditions and supplemented with recombinant murine G-CSF. Thus, G-CSF facilitates the development of clonogenic osteoclast progenitors from hematopoietic stem cells in lymphoid long-term culture conditions. This culture system may serve as a useful model for ex vivo generation of osteoclast progenitors.

Animals↗

Similarities in long-term cultures of blood and bone marrow from patients with acute myelogenous leukemia.

Dexter-type long-term cultures (LTC) were initiated with peripheral blood (PB) and/or bone marrow cells from 11 patients with acute myelogenous leukemia (AML), and 2 with myelodysplastic syndrome in blastic transformation. Marrow and PB cells from normal subjects served as controls. Assessment of nucleated cells and clonogenic progenitors in the adherent and nonadherent fractions of LTC revealed active hemopoiesis for greater than 5 wks in 4 of 8 cultures of AML blood, and 4 of 7 of AML marrow. The morphology and kinetics of nucleated cells and progenitors with putative normal (granulocyte-macrophage colony-forming units or CFU-gm), and abnormal (blast) phenotype in LTC from AML blood were similar to those from AML marrow, and adherent cells positive for collagen I and III and vimentin were found in both types of LTC. Growth of CFU-gm colonies ceased by wk 5-8 in AML cultures, significantly earlier than in LTC of normal marrow cells (survival of greater than 10 wks), which may indicate derivation of the CFU-gm from a transformed clone or deficiency of stromal function in the leukemic state. In most AML blood and AML marrow LTCs, growth of abnormal (blast) colonies continued until wk 4-6. This study demonstrates certain similarities of morphology and function between LTC of AML blood and AML marrow cells. LTC may provide a useful model for further analysis of circulating primitive hemopoietic progenitor cells in leukemic states.

Blood Cells↗

Hematopoiesis on cellulose ester membranes (CEM). IX. Enrichment of membranes with adherent layers from long-term marrow culture from normal or drug-treated mice (hematopoietic microenvironment study II).

Cellulose ester membranes (Millipore) or polytetrafluoroethylene (Mitex) membranes were coated with adherent layers taken from Dexter-type long-term cultures, 4-5, 8 or 12 weeks after initiation of culture. The cultures were established with marrow taken from untreated mice or, in some cases, from mice treated with a single lethal dose (LD10) of carmustine (BCNU) or cyclophosphamide. In the studies using untreated mice, the cultures went for 8 or 12 weeks and in the drug studies, for 4-5 weeks. The 8 and 12 week cultures were reseeded at 4 weeks. The membranes were implanted into the peritoneal cavities of mice for 3-12 months after which they were removed, fixed, sectioned and stained for histologic study. After 6 months of implantation, about 40% of the membranes coated with cells from non-drug-treated mice and 60% of the membranes coated with cells from drug-treated mice contained hematopoietic elements; often there were foci of trilineal hematopoiesis. Hematopoiesis never occurred without bone formation, but the reverse was not true. Membranes coated with adherent layers established from marrow of mice treated with cyclophosphamide or BCNU showed two main characteristics: 1) they supported hematopoiesis normally, and 2) the regeneration of stroma and hematopoiesis occurred earlier than in membranes coated with stroma derived from normal mice, perhaps because the cells from the drug-treated mice spent a shorter time in culture. In vitro culture may damage cells required to condition the membrane for hematopoiesis.

Animals↗

Murine acute leukemia cell line with megakaryocytic differentiation (MK-8057) induced by whole-body irradiation in C3H/He mice: cytological properties and kinetics of its leukemic stem cells.

Five cases of murine leukemia with megakaryocytic differentiation were observed among the 417 cases of radiation-induced leukemias which developed in 30% of C3H/HeMs mice exposed at 8 to 10 weeks to 0.5 to 5 gy total body irradiation. Cells from individual leukemic colonies in the spleen of the irradiated mice, and cells from colonies in methylcellulose (MC) culture in vitro, derived from one of these leukemias, MK-8057, were injected into mice; both types of cells caused the deaths of the recipient mice by inducing the same type of leukemia. MK-8057 can be maintained in Dexter-type liquid culture with a feeder layer of irradiated bone marrow cells. There was a linear reciprocal relationship between the increasing number of MK-8057 cells injected versus the survival of the recipient mice. A reciprocal relationship also was seen between an increasing number of leukemic stem cells, corresponding to the number of MK-8057 cells, and the survival of mice injected with MK-8057. Giant nuclear megakaryocytes developed during the course of colony growth in the spleen as they did in the MC culture. Such megakaryocytes were acetylcholinesterase positive, whereas leukemic cells in the peripheral blood showed no sign of platelet production nor of a positive reaction to acetylcholinesterase. Cells maintained in culture were entirely positive in platelet glycoprotein IIb/IIIa when anti-human antibody was used. The larger cells in a splenic cell suspension derived from a moribund mouse were separated and enriched by velocity sedimentation using centrifugal elutriation (CE), and then subjected to flow cytometry using propidium iodide staining. Cells with up to 32N-DNA content were detected. After separating MK-8057 by counter-flow CE, the larger cell fraction (mode at 540 microns3) produced more leukemic colonies when injected into irradiated mice than did the small cell fraction (mode at 240 microns3). A higher percent of the larger cell fraction (61.9%) was killed by the addition of tritiated thymidine cytocide than in the smaller cell fraction (14.9%). Thus, the smaller cell fraction is considered to have more leukemic spleen colony-forming units (L-CFU-s) in the resting state.

Animals↗

Myelosuppressive effects of cytosine arabinoside (Ara-C) on growth factor-dependent human long-term bone marrow cultures (LTBMC).

Freshly isolated human mononuclear cells (5 x 10(6)) were incubated in a Dexter-type long-term bone marrow culture (LTBMC) system to study myelosuppressive effects of cytosine arabinoside (Ara-C) in combination with granulocyte-macrophage colony-stimulating factor (GM-CSF) or interleukin 3 (IL-3). Differential counts (dc) of the nonadherent cell (nac) populations, starting with culture initiation, were performed weekly. After one week of simultaneous incubation of LTBMCs with either cytokine (100 ng/ml) and Ara-C (1 microgram/ml), nac numbers were markedly reduced compared to controls. Dc after week 1 of culture demonstrated significant decreases of all myeloid cell fractions except for macrophages, which remained unaffected. Growth factor-dependent LTBMCs exposed to Ara-C showed recovery of promyelocytic, metamyelocytic, and polymorphonuclear cell numbers up to control values (cultures without Ara-C exposure) in weeks 3 to 6. Intriguingly, high-proliferative, early myeloid progenitor cells (myeloblasts) appeared at high rates only in IL-3-dependent LTBMCs with and without Ara-C exposure. Nac numbers in LTBMCs exposed to Ara-C alone declined rapidly; after two weeks of culture only negligible numbers of viable nac were maintained. Plating experiments of nac in the presence of GM-CSF were performed weekly. Granulocyte-macrophage colony-forming units (CFU-GM) yields for nac from IL-3 LTBMCs were consistently higher than those for nac from GM-CSF LTBMCs. Ara-C exposure reduced CFU-GM numbers generated with nac from GM-CSF LTBMCs to 10% of GM-CSF controls (week 1). However, CFU-GM numbers grown with nac from Ara-C exposed GM-CSF-dependent LTBMCs recovered above control levels after week 3.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Enhancement of cell production in long-term bone marrow culture.

In an effort to increase the long-term production of hematopoietic cells in vitro, Origen hybridoma cloning factor (HCF) was added at the initiation of Dexter type cultures, in which whole bone marrow (BM) was seeded into tissue culture flasks and formed an adherent stromal layer that supported the proliferation and differentiation of primitive cells. After about six weeks, all the cultures were fully established, and continuous production of nonadherent cells was maintained for at least 27 weeks. In the groups with 20% HCF, there was a significant (three- to fourfold) increase in the steady-state cell production of 106 +/- 17 x 10(4) cells/ml compared to 26 +/- 10 x 10(4) in controls. In some cases the ability of HCF to increase productivity was limited by the nutrients and metabolic products in the culture medium. Cell number varied inversely with glucose and pH. HCF increased the concentration and absolute number of myeloid progenitors (granulocyte-macrophage colony forming units and spleen colony forming units) in the nonadherent layer and shifted the differentiation of granulocyte-macrophage colony forming units toward the production of cells of the monocyte/macrophage lineage. Spleen colonies produced from 10(5) cells from cultures with HCF were more numerous (8 +/- 2 versus 4 +/- 2) and larger than those from control cultures (2.6 versus 0.2 mg/colony), but they contained the usual cell lineages (erythrocytic, granulocytic and megakaryocytic).

Animals↗

Can human hematopoietic stem cells be cultured ex vivo?

The factors that induce proliferation of the human hematopoietic stem cell are ill defined. Further characterization of such growth factors will be needed to develop ex vivo culture systems that induce prolonged proliferation and expansion of human hematopoietic stem cells. Human or murine hematopoietic progenitors that can initiate and sustain long-term culture systems (LTC-IC) represent a population of very primitive hematopoietic progenitors. When cultured in direct contact with stromal layers, we and others have demonstrated that a fraction of such LTC-IC can be maintained. In addition, stroma-free long-term cultures supplemented with two to nine cytokines can induce proliferation and differentiation of immature human hematopoietic progenitors. However, 70-90% of primitive LTC-IC are lost after five weeks in such cultures. We describe a "stroma-non-contact" culture system, in which progenitors are cultured separated from stroma by a 0.4 micron microporous membrane which prevents cell stroma contact but allows free passage of diffusible factors. Primitive progenitors in such cultures can not only differentiate into committed progenitors but also are maintained to a greater extent than in Dexter cultures. We will discuss the relative contribution of 1) direct contact between hematopoietic progenitors and bone marrow stroma, 2) soluble stroma-derived factors and 3) previously characterized growth promoting and presumed growth inhibitory cytokines in the in vitro maintenance and potential expansion of LTC-IC.

Animals↗

Interleukin 4 alters human bone marrow stroma and modulates its interaction with hematopoietic progenitors.

To investigate the functional activity of interleukin 4 (IL-4) on human marrow stroma formation, normal bone marrow (BM) samples were cultured in "Dexter-type" long-term cultures in the presence and absence of IL-4. IL-4 (0.001 to 1.0 micrograms/ml) added at the initiation of culture and once weekly when the cultures were fed effaced the culture architecture. In four-week old confluent cultures smooth muscle-like and endothelial-like cells were rare, the fibronectin network and cobblestone areas were absent, and a preponderance of monocyte-macrophages characterized the adherent layer. Exposure to IL-4 reduced the numbers of CD34+ cells, colony-forming unit granulocyte-macrophage (GFU-GM) cells and burst-forming unit-erythroid (BFU-E) cells in the adherent layer, and increased their numbers in the nonadherent layer. In five of eight IL-4-containing cultures the concentrations of macrophage colony-stimulating factor (M-CSF) were increased and in two of eight IL-4-treated cultures the concentrations of tumor necrosis factor-alpha (TNF-alpha) were significantly elevated as compared to those in control cultures, whereas there were no consistent differences in the levels of either IL-6 or transforming growth factor-beta (TGF-beta). IL-1 beta and granulocyte-macrophage CSF (GM-CSF) were not detected in any culture. These data suggest that IL-4 suppresses stroma formation and alters its structure and cellular composition.

Antigens, CD↗

Capuchin monkeys, Cebus apella fail to understand a cooperative task

We investigated whether capuchin monkeys cooperate to solve a task and to what extent they take into account the behaviour of another individual when cooperating. Two groups of capuchin monkeys (N=5 and 6) were tested in a task whose solution required simultaneous pulling of two handles which were too far from one another to be pulled by one monkey. Before carrying out the cooperation study, individual monkeys were trained to pull one handle (training phase 1) and to pull two handles simultaneously (training phase 2) for a food reward. Nine subjects were successful in training phase 1, and five in training phase 2. In the cooperation study seven subjects were successful, that is, pulled one handle while a companion pulled the other. Further analyses revealed that capuchins did not increase their pulling actions when a partner was close to or at the other handle, that is, when cooperation might occur. These data suggest that capuchin monkeys acted together at the task and got the reward without understanding the role of the partner and without taking its behaviour into consideration. Social tolerance, as well as their tendency to explore and their manual dexterity, were the major factors accounting for the capuchins' success.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour

Journal Article↗