Proliferation dynamics of acute myeloid leukaemia and haematopoietic progenitors competing for bone marrow space.

Leukaemia progressively invades bone marrow (BM), outcompeting healthy haematopoiesis by mechanisms that are not fully understood. Combining cell number measurements with a short-timescale dual pulse labelling method, we simultaneously determine the proliferation dynamics of primitive haematopoietic compartments and acute myeloid leukaemia (AML). We observe an unchanging proportion of AML cells entering S phase per hour throughout disease progression, with substantial BM egress at high levels of infiltration. For healthy haematopoiesis, we find haematopoietic stem cells (HSCs) make a significant contribution to cell production, but we phenotypically identify a quiescent subpopulation with enhanced engraftment ability. During AML progression, we observe that multipotent progenitors maintain a constant proportion entering S phase per hour, despite a dramatic decrease in the overall population size. Primitive populations are lost from BM with kinetics that are consistent with ousting irrespective of cell cycle state, with the exception of the quiescent HSC subpopulation, which is more resistant to elimination.

T-cell stimuli independently sum to regulate an inherited clonal division fate.

In the presence of antigen and costimulation, T cells undergo a characteristic response of expansion, cessation and contraction. Previous studies have revealed that population-level reproducibility is a consequence of multiple clones exhibiting considerable disparity in burst size, highlighting the requirement for single-cell information in understanding T-cell fate regulation. Here we show that individual T-cell clones resulting from controlled stimulation in vitro are strongly lineage imprinted with highly correlated expansion fates. Progeny from clonal families cease dividing in the same or adjacent generations, with inter-clonal variation producing burst-size diversity. The effects of costimulatory signals on individual clones sum together with stochastic independence; therefore, the net effect across multiple clones produces consistent, but heterogeneous population responses. These data demonstrate that substantial clonal heterogeneity arises through differences in experience of clonal progenitors, either through stochastic antigen interaction or by differences in initial receptor sensitivities.

Development of cytochrome oxidase blobs in visual cortex of normal and visually deprived cats.

Cytochrome oxidase (CO) blobs are central to our understanding of the columnar organization and parallel pathways in primate and cat visual cortex. In primates, development of blobs and their relationship with other columnar features of the visual cortex begins pre-natally, before visual experience. In kittens, the supragranular layers differentiate post-natally, after eye opening, raising the possibility that visual experience may influence the development of blobs in cat V1. We have examined the development of blobs in unfolded and flattened sections through the visual cortex of normally reared, dark-reared, monocularly deprived and binocularly deprived kittens. Blobs were found in superficial layers of V1 of normally reared kittens as early as 2 weeks of age, although at this age the overall CO staining in V1 was lighter than in V2. By 6 weeks of age the blobs were adult-like. A patchy pattern of CO staining was also found in V2 of young kittens but not in adults. Visual experience was not necessary for expression of the blobs and monocularly deprived kittens had well developed blobs, indicating that strong Y cell drive is not necessary for the development of blobs in cat V1. CO blobs appear in kitten V1 very early in post-natal development and their expression is independent of visual experience, suggesting that they may be an intrinsic feature of V1 organization.

Experience-dependent development of NMDAR1 subunit expression in the lateral geniculate nucleus.

Monocular deprivation early in postnatal development leads to anatomical and physiological changes in the lateral geniculate nucleus (LGN) and visual cortex. Many of these changes are dependent upon activation of the NMDA receptor. We have examined the role of visual experience in modifying NMDAR1 subunit expression in the LGN of animals reared with various forms of visual deprivation. Following monocular deprivation initiated either at eye opening or at the peak of the critical period, there were approximately 20% fewer NMDAR1-immunopositive neurons in the deprived laminae of the LGN. The loss of NMDAR1-immunopositive neurons was found throughout both the binocular and monocular segments of the LGN and after monocular deprivation until just 3 weeks of age. These results indicate that the loss of NMDAR1 in the LGN following monocular deprivation does not simply reflect changes in the visual cortex. The loss of NMDAR1 expression was not necessarily permanent. Initiation of binocular vision at the peak of the critical period ameliorated the effect of monocular deprivation and the introduction of a period of reverse occlusion led to a complete reversal. Taken together, the results show that the expression of the NMDAR1 subunit in the LGN can be modified by the pattern of visual experience during postnatal development.

Analysis of the postnatal growth of visual cortex.

Development and growth of V1 begins during embryogenesis and continues postnatally. The growth of V1 has direct implications on the organization of features such as the retinotopic map and the pattern of visual cortical columns. We have examined the postnatal growth and two-dimensional shape of V1 in macaque monkeys, cats, and rats. The perimeter, area, and anterior-posterior length of V1 were measured from unfolded and flattened sections from neonatal and adult animals from each of these species. Although there were substantial differences in the overall amount of postnatal growth, from 18% in macaque monkeys to more than 100% in cats, in all three species the shape of V1 did not change during development. Thus, growth of the mammalian visual cortex is well described as an isotropic expansion, so the layout of the global features, such as the arrangement of ocular dominance columns and the retinotopic map, does not need to change during development. Furthermore, quantification of the shape confirms the observations that there is a similar, egg-like oval shape to the visual cortex of these mammalian species.

Patchy distribution of NMDAR1 subunit immunoreactivity in developing visual cortex.

Development of ocular dominance columns is dependent on patterned retinal activity, and yet patterned activity alone cannot explain all aspects of cortical column development. Features intrinsic to the cortex have been proposed to interact with activity to guide the patterning of cortical columns (), and the NMDA receptor, because of its role in experience-dependent plasticity, is an obvious candidate. Using immunohistochemical techniques, we found a transiently patchy distribution of the NMDA receptor 1 (NMDAR1) subunit in kitten visual cortex. Regularly spaced patches of NMDAR1-immunoreactive neurons were found at the top of the cortical plate in the developing visual cortex at 2 weeks of age. At 4-5 weeks of age, the radial extent of the NMDAR1 patches spanned the supragranular layers, and by 12 weeks of age, this nonuniform pattern of NMDAR1 immunostaining was no longer apparent. Monocular visual experience prevented the expression of the NMDAR1 patches, but just 4 d of subsequent binocular visual experience was sufficient to promote expression of the patches. Furthermore, the NMDAR1 patches tended to be associated with the borders of ocular dominance columns. These results suggest that the degree of plasticity associated with NMDA-mediated mechanisms is elevated in local regions across the tangential extent of the visual cortex and that the NMDAR1 patches may participate in sculpting the overall arrangement of visual cortical columns.

Secretory and nonsecretory pituitary adenomas are distinguishable by 1/T1 magnetic relaxation rates at very low magnetic fields in vitro.

RATIONALE AND OBJECTIVES: The authors investigated whether hormonally active and inactive pituitary adenomas can be discriminated in vitro by magnetic resonance (MR) imaging-related data. METHODS: 1/T1 nuclear magnetic relaxation dispersion profiles were measured for 39 fresh surgical specimens of secreting and nonsecreting adenomas, classified using clinical criteria or preoperative serum hormone levels. Nonsecreting adenomas were subdivided into hormone-producing and nonhormone-producing by immunostains. At five fields (0.00024 to 1.2 tesla [T]), mean 1/T1 was analyzed for statistically significant differences among these three tumor categories. RESULTS: Mean 1/T1 was significantly higher (P < 0.02) for hormone-secreting than for nonsecreting adenomas at fields below 0.24 T; no significant difference existed at typical MR imaging fields (0.5 to 1.5 T). Mean 1/T1 for hormone-producing and nonhormone-producing, nonsecreting adenomas were not significantly different at any field. CONCLUSIONS: Because 1/T1 at low fields is related to 1/T2 at imaging fields, it may be possible to detect hormone secretion of pituitary adenomas noninvasively by MR imaging.

Relaxometry of noncalcified human meningiomas. Correlation with histology and solids content.

RATIONALE AND OBJECTIVES: Resected meningiomas were examined by relaxometry and light microscopy to evaluate the potential of magnetic resonance imaging (MRI) for identifying histologic subtypes and for discriminating among benign, radiation therapy-induced, and malignant meningiomas. METHODS: The magnetic field dependence of 1/T1 of water protons (nuclear magnetic relaxation dispersion [NMRD] profile) and the water content (dry weight) were measured for 67 specimens, and the data were compared with histology. Only noncalcified, nonhemorrhagic meningiomas are reported. RESULTS: No correlations were found between NMRD profiles, dry weight, and any histologic subtype, in contrast to an analogous study of astrocytomas. Rather, meningiomas have a broader variability of dry weight and 1/T1 than related parenchyma but a much narrower range than all grades of astrocytomas. The mean value of 1/T1, at all fields, is slightly higher in meningiomas--and the mean water content about the same--as adult cortical gray matter. CONCLUSION: Meningiomas are frequently isointense with cortex, and histologic subtypes cannot be differentiated at any magnetic field strength by MRI using only T1- or proton density-weighted MRI.

Multiple spinal intradural schwannomas in the absence of von Recklinghausen's disease.

A case of multiple spinal schwannomas is reported without any evidence of other manifestations of neurofibromatosis. Although cases of multiple schwannomas have been described in the setting of "Schwannomatosis," this case is unique in that all tumors were within the spinal canal. The presence of a schwannoma should prompt a complete investigation for other tumors of the nervous system preferably with magnetic resonance imaging.

Intramedullary spinal cord abscess.

Viral myelitis and bacterial epidural infections are common in intravenous drug abusers, but primary infections of the spinal cord are extremely rare. We report a 50-year-old active intravenous drug user who developed tetraplegia from an intramedullary abscess caused by Pseudomonas cepacia. Despite neurosurgical drainage and appropriate antibiotic therapy, no improvement was seen. Earlier intervention and a high index of suspicion is required in patients with a history of intravenous drug abuse and spinal cord symptoms.

Human blood-brain barrier insulin-like growth factor receptor.

Insulin-like growth factor (IGF)-1 and IGF-2, may be important regulatory molecules in the CNS. Possible origins of IGFs in brain include either de novo synthesis or transport of circulating IGFs from blood into brain via receptor mediated transcytosis mechanisms at the brain capillary endothelial wall, ie, the blood-brain barrier (BBB). In the present studies, isolated human brain capillaries are used as an in vitro model system of the human BBB and the characteristics of IGF-1 or IGF-2 binding to this preparation were assessed. The total binding of IGF-2 at 37 degrees C exceeded 130% per mg protein and was threefold greater than the total binding for IGF-1. However, at 37 degrees C nonsaturable binding equaled total binding, suggesting that endocytosis is rate limiting at physiologic temperatures. Binding studies performed at 4 degrees C slowed endocytosis to a greater extent than membrane binding, and specific binding of either IGF-1 or IGF-2 was detectable. Scatchard plots for either peptide were linear and the molar dissociation constant of IGF-1 and IGF-2 binding was 2.1 +/- 0.4 and 1.1 +/- 0.1 nmol/L, respectively. Superphysiologic concentrations of porcine insulin inhibited the binding of both IGF-1 (ED50 = 2 micrograms/mL) and IGF-2 (ED50 = 0.5 microgram/mL). Affinity cross linking of 125I-IGF-1, 125I-IGF-2, and 125I-insulin to isolated human brain capillaries was performed using disuccinimidylsuberate (DSS). These studies revealed a 141 kd binding site for both IGF-1 and IGF-2, and a 133 kd binding site for insulin.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood-brain barrier transcytosis of insulin in developing rabbits.

Previous studies with isolated brain microvessels have suggested that blood insulin is selectively transported through the brain capillary, i.e. the blood-brain barrier (BBB), by receptor-mediated transcytosis. The purpose of the present study is to demonstrate in vivo the uptake of circulating 125I-insulin by brain using thaw-mount autoradiography. However, metabolism of systemic 125I-insulin to 125I-tyrosine would allow for brain uptake of 125I-tyrosine and this would preclude interpretation of the autoradiogram. Therefore, the present studies were performed in developing rabbits, since plasma protein degradation of peptides is greatly reduced in developing animals. 125I-insulin was infused via the carotid artery at a rate of 0.25 ml/min for 1, 5, or 10 min, and the mean brain uptake, relative to a [3H]albumin reference, was 99.3 +/- 5.5%, 110.1 +/- 4.3%, and 143.6 +/- 7.9%, respectively. This uptake was saturable by simultaneously infusing unlabeled insulin. Thaw-mount autoradiography of rabbit brain after a 10-min infusion of 125I-insulin revealed silver grains in the pericapillary space and well within the brain parenchyma. HPLC analysis of acid-ethanol extracts of rabbit blood after a 10-min infusion showed virtually all of the 125I-radioactivity co-migrated with a known insulin standard on a reverse-phase column, indicating minimal degradation of infused 125I-insulin. HPLC analysis of brain radioactivity showed the major peak co-migrated with 125I-insulin and this peak was precipitated by an anti-insulin antiserum. The correlation of the transport data, the autoradiography, and the HPLC analysis support the model that brain insulin originates from blood via receptor-mediated transport of the peptide at the BBB.

LES pressure response to pentagastrin: effect of cholinergic augmentation and inhibition.

Studies were performed on the anesthetized adult opossum to examine a) the pressure response of the lower esophageal sphincter (LES) to both bolus injections and continuous infusions of pentagastrin (PG), and b) how this response is affected by either cholinergic antagonism with atropine or cholinergic enhancement with edrophonium, an anticholinesterase. Both the bolus injection and the continuous infusion of PG produced dose-dependent rises in LES pressure. The peak pressure responses occurred at 1 microgram/kg and 32 microgram/kg-h, respectively. Atropine (100 microgram/kg), in a dose that significantly diminished the LES response to exogenous acetylcholine, had no significant effect on PG-induced increases in LES pressure. Similarly, edrophonium (100 microgram/kg) had no effect on the response of the LES to PG stimulation. The Sphincter's response to exogenous acetylcholine was significantly enhanced after edrophonium pretreatment. From these studied, we conclude that PG-induced increases in the LES pressure are due primarily to the direct stimulation of sphincter smooth muscle rather than to activation of excitatory cholinergic neurons.