Evidence for spatial regularity among retinal ganglion cells that project to the accessory optic system in a frog, a reptile, a bird, and a mammal.

The vertebrate retina contains only five major neuronal classes but these embrace a great diversity of discrete types, many of them hard to define by classical methods. Consideration of their spatial distributions (mosaics) has allowed new types, including large ganglion cells, to be resolved across a wide range of vertebrates. However, one category of large ganglion cells has seemed refractory to mosaic analysis: those that project to the accessory optic system (AOS) and serve vestibulocerebellar mechanisms of motion detection and image stabilization. Whenever AOS-projecting cells have been analyzed by nearest-neighbor methods, their distribution has appeared almost random. This is puzzling, because most aspects of visual processing require the visual scene to be sampled regularly. Here, spatial correlogram methods are applied to distributions of large ganglion cells, labeled retrogradely from the AOS in frogs, turtles, and rats, and to the AOS-projecting displaced ganglion cells of chickens. These methods reveal hidden spatial order among AOS-projecting populations, of a form that can be simulated either by superimposing a single regular mosaic on a random population or, more interestingly, by overlapping three or more regular, similar but spatially independent mosaics. The rabbit is known to have direction-selective ganglion cells (not, however, AOS projecting) that can be subdivided into functionally distinct, regular mosaics by their tracer-coupling patterns even though they are morphologically homogeneous. The present results imply that the direction-selective AOS-projecting ganglion cells of all vertebrates may, likewise, be subdivided into regular, independent mosaics.

Pre-clinical and clinical study of QC12, a water-soluble, pro-drug of quercetin.

BACKGROUND: Quercetin is a naturally occurring flavonoid with many biological activities including inhibition of a number of tyrosine kinases. A phase I, dose-escalation trial of quercetin defined the maximum tolerated dose (MTD) as 1700 mg/m2 three weekly, but the vehicle, dimethyl sulphoxide (DMSO) is unsuitable for further clinical development of quercetin. PATIENTS AND METHODS: A water-soluble, pro-drug of quercetin (3'(N-carboxymethyl)carbomyl-3,4',5,7-tetrahydroxyflavone), QC12 has been synthesised. Six cancer patients received 400 mg of QC12 (equivalent to 298 mg of quercetin), orally on day 1 and intravenously (i.v.) in normal saline on day 14. RESULTS: Following oral administration of QC12 we were unable to detect QC12 or quercetin in plasma. After i.v. administration, we detected peak plasma concentrations of QC12 of 108.7 +/- 41.67 microMolar (microM). A two-compartment model with mean t(1/2)alpha of 0.31 +/- 0.27 hours and mean t(1/2)beta of 0.86 +/- 0.78 hours best described the concentration-time curves for QC12. The mean AUC was 44.54 +/- 13.0 microM.hour and mean volume of distribution (Vd) of 10.0 +/- 6.2 litres (l). Quercetin was found in all patients following i.v. infusion of QC12, with peak levels of quercetin 19.9 +/- 11.8 microM. The relative bioavailability of quercetin was estimated to be 20%-25% quercetin released from QC12. CONCLUSIONS: QC12 is not orally bioavailable. This water-soluble pro-drug warrants further clinical investigation; starting with a formal phase I, IV, dose-escalation study.

Analysis of lymphocyte diversity in the elderly: heteroduplex analysis and alternative techniques.

Heteroduplex analysis allows global analysis of T cell receptor clonality. This review outlines the method, compares it to other available techniques for the study of clonality and reviews current literature on how these are being used to investigate alterations in the T cell repertoire within elderly individuals.

Retinal mosaics: new insights into an old concept.

It has been known since the middle of the 19th century that different neuronal types are distributed across the retinal surface in non-random arrays: indeed, these arrays, called 'mosaics', have long been considered to be a fundamental feature of retinal organization. However, until recently, little was known about how such mosaics are established during development. In the hope of stimulating further research, this article reviews the current status of three very different approaches to this intriguing general problem. The first postulates arrays of molecular markers, which are produced by specific cell types shortly after their final mitotic divisions and could be influential in the differentiation of other cell types. The second invokes a tangential dispersion of differentiating cells to generate spatial order, either while these cells are still migrating or soon after they reach their laminar destinations. The third involves the elimination of wrongly positioned cells through the process of naturally occurring cell death.

Large retinal ganglion cells that form independent, regular mosaics in the bufonoid frogs Bufo marinus and Litoria moorei.

Population-based methods were used to study labeled retinal ganglion cells from the cane toad Bufo marinus and the treefrog Litoria moorei, two visually competent bufonoid neobatrachians with contrasting habitats. In both, cells with large somata and thick dendrites formed distinct types with independent mosaics. The alpha(a), alpha(ab), and alpha(c) mosaics of Bufo in all major respects resembled those of ranids, studied previously, and could be provisionally matched to the same functional classes. As in other frogs, some alpha(a) cells were displaced and many alpha-cells of all types were asymmetric, but within each type all variants belonged to one mosaic. Nearest-neighbor analyses and spatial correlograms confirmed that all three mosaics were regular and independent. In Litoria, monostratified alpha(a) cells were not found. Instead, two bistratified types were present, distinguished individually by soma size and dendritic caliber and collectively by membership of independent mosaics: the larger (approximately 0.8% of all ganglion cells) was termed alpha1(ab) and the smaller (approximately 2.2%) alpha2ab. An alpha(c) cell type was also present, although too inconstantly labeled for mosaic analysis. Nearest-neighbor analyses and spatial correlograms confirmed that the two alpha(ab) mosaics were regular and independent. Densities, proportions, soma sizes, and mosaic statistics are tabulated for each species. The emergence of a consensus pattern of alpha-cell types in fishes and frogs, from which this treefrog partly diverges, offers new possibilities for studying correlations between function, phylogeny, ecology, and neuronal form.

Mitomycin, ifosfamide, and cisplatin in unresectable non-small-cell lung cancer: effects on survival and quality of life.

PURPOSE: Chemotherapy for non-small-cell lung cancer (NSCLC) remains controversial. We describe the two largest reported, randomized, parallel trials designed to determine whether the addition of chemotherapy influences duration and quality of life in localized, unresectable (mitomycin, ifosfamide, cisplatin [MIC]1 trial) and extensive (MIC2 trial) disease. PATIENTS AND METHODS: Ambulatory patients with NSCLC, aged 75 years or younger, with localized disease, were randomized in MIC1 to receive up to four cycles of chemotherapy (CT: mitomycin 6 mg/m(2), ifosfamide 3 g/m(2), and cisplatin 50 mg/m(2)) every 21 days, followed by radical radiotherapy (CT + RT) or radiotherapy (RT) alone. Extensive-stage patients were randomized in MIC2 to identical chemotherapy plus palliative care (CT + PC) or palliative care (PC) alone. Short-term change in quality of life (QOL) was assessed in a subgroup of patients. Data from the two trials were combined to allow multivariate and stratified survival analyses. RESULTS: Seven hundred ninety-seven eligible patients were randomized, 446 in MIC1 and 351 in MIC2. MIC CT improved survival in both trials (significantly in MIC2). The median survival time in MIC1 was 11.7 months (CT + RT) versus 9.7 months (RT alone) (P =.14); whereas in MIC2, median survival time was 6.7 months (CT + PC) compared with 4. 8 months (PC alone) (P =.03). QOL, assessed in 134 patients from start of trial to week 6, showed improvement with chemotherapy and deterioration with standard treatment. In the combined analysis of 797 randomized patients, the positive effect of MIC on survival was significant overall (P =.01) and after adjusting for prognostic factors (P =.01). CONCLUSION: MIC chemotherapy prolongs survival in unresectable NSCLC without compromising QOL.

Neuronal and glial cell types revealed by NADPH-diaphorase histochemistry in the retina of a teleost fish, the grass goby (Zosterisessor ophiocephalus, Perciformes, Gobiidae).

The grass goby is a mud-burrowing fish with a rich retinal vasculature appropriate to its hypoxic habitat. NADPH-diaphorase histochemistry was performed on retinal sections and wholemounts to reveal cells that contain nitric oxide synthase and so may be presumed to synthesise nitric oxide, a gaseous intercellular messenger with many roles including vasodilation. Structures that were consistently stained by this method included cone ellipsoids, horizontal cells, Müller cells and their processes, large displaced ganglion cells in the inner nuclear layer (identified by their axons), large interstitial ganglion cells in the inner plexiform layer, and capillary endothelial cells. In wholemounts, horizontal cells were seen to form a regular pattern, contacting each other at their dendritic terminals. Some cells in the ganglion cell layer were weakly stained, but stained bipolar and amacrine cells were not seen. The diaphorase-positive large ganglion cells all formed large, sparsely branched dendritic trees, arborizing near the scleral border of the inner plexiform layer. The displaced and interstitial cells seemed to belong to distinct morphological types, the interstitial cells having smaller somata and trees. Analysis of their spatial distributions in one representative retina confirmed this: the displaced cells formed a highly regular mosaic with a mean spacing (nearest-neighbour distance) of 303 microm, whereas the interstitial cells formed a separate mosaic, almost as regular but with a smaller mean spacing of 193 microm, rising to 217 microm in a sample that excluded the area retinae temporalis. Spatial correlogram analysis showed that these two mosaics were spatially independent. Nitric oxide probably has many roles in the retina. The presence of its synthetic enzyme in Müller cells, which communicate with retinal blood vessels, is consistent with a role in the control of retinal blood flow. Its function in large, mosaic-forming retinal ganglion cells is unknown.

Species-dependent variation in the dendritic stratification of apparently homologous retinal ganglion cell mosaics in two neoteleost fishes.

Large retinal ganglion cells of the marine neoteleost Bathymaster derjugini were labeled with horseradish peroxidase and studied in flatmounts. Four types formed regular, independent mosaics, of which three (biplexiform, alpha-a, alpha-c) resembled those in several other teleosts. The fourth (alpha-ab) appeared novel in one significant respect. Whereas we originally described similar cells in another neoteleost, Oreochromis spilurus, as monostratified in sublamina b of the inner plexiform layer, these were very clearly bistratified in a and b. Detailed re-analysis of our Oreochromis flatmounts showed that the difference is of one degree only: many Oreochromis cells do send fine dendrites into a. These observations strengthen the evidence that all four mosaics are homologous across a wide range of fishes, and clear away an obstacle to our earlier proposals that the alpha-a, alpha-ab and alpha-c mosaics of fishes, frogs, and perhaps other nonmammalian jawed vertebrates too, may all be homologous.

Somatic and dendritic mosaics formed by large ganglion cells in the retina of the common house gecko (Hemidactylus frenatus).

Recent studies of large ganglion cells in fishes and frogs have identified a shared inventory of three basic types, with characteristic forms and spatially independent mosaic distributions. These anamniote types and mosaics are hard to match to the large ganglion cell types and mosaics of mammals, implying that the underlying developmental programmes have diverged during evolution. Reptiles and mammals both belong to the amniote lineage, so the point of divergence can be investigated by comparing the large ganglion cells of reptiles with those of mammals, taking fishes and frogs as outgroups. With this aim, ganglion cells of the common house gecko, Hemidactylus frenatus, were labelled with horse-radish peroxidase by an in vitro method and studied in retinal flatmounts. Two prominent, regular, spatially independent mosaics were consistently present. One (alpha a) was characterized by somata displaced into the inner nuclear layer and dendrites forming planar trees in sublamina a; the other (alpha ab) comprised large orthotopic somata and distinctive, bistratified dendrites that formed discrete planar subtrees in sublaminae a and b. These subtrees were joined by up to 40 vertical link segments, whose distribution was found to correlate with the underlying photoreceptor mosaic. Some specimens also contained patches of a third mosaic (alpha c), characterized by large orthotopic somata and very large flat trees in sublamina c, but the labelling of this type was inconsistent. These reptilian mosaics share several distinctive characters with anamniote alpha-cell mosaics but differ markedly from the ganglion cell mosaics of any known mammal. The most parsimonious conclusion is that those mosaic features that are shared by the ganglion cells of all nonmammals are homologous and primitive (symplesiomorphic), while those that are shared by all therian mammals are homologous and derived (synapomorphic). This is consistent with other differences between mammalian and nonmammalian eyes. Mosaic formation itself, however, seems to be a universal characteristic of large ganglion cells.

Expression of major gap junction connexin types in the working myocardium of eight chordates.

The alpha1 connexin (connexin43) is regarded as the major gap junction protein of the myocardium because it predominates there in mammals. Here, we show that it is not the major connexin of the working myocardium in non-mammalian vertebrates, which instead express beta1-like connexins homologous to mammalian connexin32. A phylogenetic series of hearts was immunostained with seven antibodies raised against peptide sequences specific for three distinct members of the gap junction connexin family: alpha1, beta1 and alpha5 (mammalian connexin40/avian connexin42). Working myocardium from two ascidian chordates (Ciona and Mogula), a teleost (Carassius), a frog (Xenopus) and two reptiles (Anolis and Alligator) was found to express a beta1-like connexin, rather than an alpha1-like connexin. An alpha1-like connexin was nevertheless often detected in other cardiac tissues. In the chicken (by ancestry a reptile), the developing myocardium expressed a beta1-like connexin strongly on embryonic day 6 but less strongly at hatching, and minimally in the adult. Myocardial expression of alpha5 connexin increased during development, but remained strongest in the coronary vascular endothelial and cardiac conduction tissues. The arteriolar smooth muscle of the chicken expressed alpha1 connexin throughout development, but its myocardium did not. In contrast, the working myocardium of a marsupial mammal (the opossum Trichosurus) strongly expressed an alpha1 connexin just like placental mammals. These results imply that a shift from beta1 to alpha1 connexin expression in the heart occurred prior to the evolution of the opossums. The beta and alpha connexin subfamilies have different permeabilities and gating properties, and we discuss factors that might have made this shift beneficial.

[Protein-energy malnutrition and hypobaric stress effects on ultrastructure of the heart muscle in rats]. / Efecto de la desnutrición calórico-proteica y del estrés hipóxico sobre la ultraestructura del músculo cardíaco de rata.

A model that simulated nutritional and hypoxic stress is discussed. Wistar male adult rats were used in four groups: A1, A2, B1 and B2. A1 and A2 groups were fed with a diet whose composition was: 16% protein, 43% carbohydrate, and 6% lipids. Group B1 and B2 received the same type of diet but was restricted to 70% the amount consumed by groups A1 and A2. All groups were fed for 15 days. After these 15 days, the half of both groups were introduced into an hypobaric chamber simulating an altitude of 3,500 m for two hours. Pieces of the rats hearts were used for biochemical and ultrastructural studies. In group B a dear decrease in the body and the heart weight was observed. Also a decrement of the heart proteins concentration was noticed. On the contrary the mitochondrial proteins increased. These biochemical changes correlate with the ultrastructural changes observed in the same group in which a decrease of the myofibrilles and abnormal configurations of the mitochondria were found. The undemourishment and the hypobaric stress produced an evident alteration in the ultrastructure of the heart.

Large retinal ganglion cells in the pipid frog Xenopus laevis form independent, regular mosaics resembling those of teleost fishes.

Population-based studies of retinal neurons have helped to reveal their natural types in mammals and teleost fishes. In this, the first such study in a frog, labeled ganglion cells of the mesobatrachian Xenopus laevis were examined in flatmounts. Cells with large somata and thick dendrites could be divided into three mosaic-forming types, each with its own characteristic stratification pattern. These are named alpha a, alpha ab, and alpha c, following a scheme recently used for teleosts. Cells of the alpha a mosaic (approximately 0.4% of all ganglion cells) had very large somata and trees, arborizing diffusely within sublamina a (the most sclerad). Their distal dendrites were sparsely branched but achieved consistent coverage by intersecting those of their neighbors. Displaced and orthotopic cells belonged to the same mosaic, as did cells with symmetric and asymmetric trees. Cells of the alpha ab mosaic (approximately 1.2%) had large somata, somewhat smaller trees that appeared bistratified at low magnification, and dendrites that branched extensively. Their distal dendrites arborized throughout sublamina b and the vitread part of a, tessellating with their neighbors. All were orthotopic; most were symmetric. Cells of the alpha c mosaic (approximately 0.5%) had large somata and very large, sparse, flat, overlapping trees, predominantly in sublamina c. All were orthotopic; some were asymmetric. Nearest-neighbor analyses and spatial correlograms confirmed that each mosaic was regular and independent, and that spacings were reduced in juvenile frogs. Densities, proportions, sizes, and mosaic statistics are tabulated for all three types, which are compared with types defined previously by size and symmetry in Xenopus and potentially homologous mosaic-forming types in teleosts. Our results reveal strong organizational similarities between the large ganglion cells of teleosts and frogs. They also demonstrate the value of introducing mosaic analysis at an early stage to help identify characters that are useful markers for natural types and that distinguish between within-type and between-type variation in neuronal populations.

Bone mineral changes during tibial fracture healing.

The traditional assessment of fracture healing by manipulation and viewing of radiographs is subjective and qualitative. Dual energy x-ray absorptiometry by contrast provides an accurate, precise, and minimally invasive quantitative measure of bone mineral density, a property that shows strong correlations with various mechanical properties of bone. Fourteen patients with unilateral tibial shaft fractures stabilized by external fixation were monitored with dual energy x-ray absorptiometry at monthly intervals after fracture. Fractured and contralateral unfractured bones (controls) were scanned on each occasion. Changes in mineralization with time over the whole length of the fractured bone could be seen. The most pronounced effects were visible in the area of the fracture, with a minimum recorded fracture site bone mineral density of 38 +/- 13% of contralateral values, but often more long term alterations in bone mineral density affected regions at some distance from this zone. Significantly, in four patients who had scans 5 or more months after fracture, the mineralization at the fracture site had returned to control levels, whereas bone mineral density in a region proximal to the fracture showed evidence of persisting posttraumatic osteoporosis.

TGF-beta differentially modulates epidermal growth factor-mediated increases in leukemia-inhibitory factor, IL-6, IL-1 alpha, and IL-1 beta in human thymic epithelial cells.

The regulation of cytokine production by thymic epithelial cells (TEC) in the thymus is under coordinated and temporal control and is important for the development of T cells. Human TEC express TGF-beta R and epidermal growth factor (EGF) receptor, and produce TGF-beta 3 in vitro and in vivo. Furthermore, EGF has been shown to increase IL-1 alpha, IL-1 beta, IL-6 mRNA and protein levels in human TEC. Since EGF has been shown to modulate TGF-beta effector functions, we determined whether TGF-beta can modulate EGF-mediated increases in cytokine gene expression in human TEC. We established that a single TEC expresses both EGF receptor and TGF-beta R. TGF-beta plus EGF synergistically increased leukemia-inhibitory factor (LIF), additively increased IL-6, but had little effect on IL-1 alpha and IL-1 beta mRNA levels. In contrast, TGF-beta alone increased LIF and IL-6, had little effect on IL-1 alpha, and slightly decreased IL-1 beta mRNA levels. The increases in LIF and IL-6 mRNA levels by TGF-beta plus EGF correlate with the increases in LIF and IL-6 concentrations in TEC culture supernatants as detected by ELISA. We also determined the mechanism responsible for the increases in cytokine mRNA levels. TGF-beta plus EGF did not affect transcription of LIF and IL-6 genes; this suggests that the increases in the steady state levels of cytokine mRNA were mediated post-transcriptionally, most likely at the level of mRNA stability. Our data demonstrate that TGF-beta modulates TEC cytokine production. We speculate that TGF-beta produced in situ plays a role in thymocyte development by directly affecting thymocyte differentiation and by indirectly modulating TEC cytokine production.

Large retinal ganglion cells that form independent, regular mosaics in the ranid frogs Rana esculenta and Rana pipiens.

Population-based studies of ganglion cells in retinal flatmounts have helped to reveal some of their natural types in mammals, teleost fish and, recently, the aquatic mesobatrachian frog Xenopus laevis. Here, ganglion cells of the semiterrestrial neobatrachian frogs Rana esculenta and Rana pipiens have been studied similarly. Ganglion cells with large somata and thick dendrites could again be divided into three mosaic-forming types with distinctive stratification patterns. Cell dimensions correlated inversely with density, being smallest in the visual streak. Cells of the alpha a mosaic (< 0.2% of all ganglion cells) had the largest somata at each location (often displaced) and their trees were confined to one shallow plane within sublamina a of the inner plexiform layer. In regions of high regularity, many trees were symmetric. Elsewhere, asymmetric, irregular trees predominated and their dendrites, although sparsely branched, achieved consistent coverage by intersecting in complex ways. Cells of the alpha ab mosaic were more numerous (approximately 0.7%) and had large somata, smaller (but still large) trees, and dendrites that branched extensively in two separate shallow planes in sublaminae a and b. The subtrees did not always match in symmetry, and each subtree tessellated independently with its neighbors. Cells of the alpha c mosaic (approximately 0.1%) had large, orthotopic somata and large, sparse trees (often asymmetric and irregular) close to the ganglion cell layer. Nearest-neighbor analyses and spatial correlograms confirmed that each mosaic was regular and independent. Densities, proportions, sizes, and mosaic statistics are tabulated for all three types, which are compared with types defined by size and symmetry in R. pipiens, by discriminant analysis in R. temporaria, by physiological response in both, and by mosaic analysis in Xenopus and several teleosts. The variable stratification of these otherwise similar types across species is consistent with other evidence that stratification may be determined, in part, by functional interactions.

Recombinant mitotoxin basic fibroblast growth factor-saporin reduces venous anastomotic intimal hyperplasia in the arteriovenous graft.

BACKGROUND: The plant cytotoxin saporin (SAP) is a potent ribosome-inactivating protein. When conjugated to basic fibroblast growth factor (FGF2), it selectively kills proliferating cells that have upregulated FGF receptors. In this study, we evaluated the effect of the recombinant chimeric mitotoxin rFGF2-SAP on venous anastomotic intimal hyperplasia, a major cause of failure of arteriovenous (AV) grafts. METHODS AND RESULTS: Recently designed expanded polytet-rafluoroethylene-based local infusion devices were implanted bilaterally as femoral AV conduits in six dogs. The venous anastomoses were the sites of continuous delivery of rFGF2-SAP (2.7 micrograms.kg-1.d-1) to one side and vehicle (4.6 microL.kg-1.d-1) as control to the contralateral side for 14 days. All animals survived, and all grafts were patent. Liver enzyme levels and histological analyses of liver, kidneys, and brain were normal, indicating the absence of systemic toxicity. Morphometric measurements and measurements of cell proliferation by bromodeoxyuridine index analysis were performed at both arterial and venous anastomoses. There were no significant differences between the treated grafts and the control grafts in intimal hyperplasia and intimal cell proliferation at the arterial anastomoses. In contrast, rFGF2-SAP reduced intimal thickness by 32%, intimal area by 40%, and cell proliferation index by 33% at the treated venous anastomoses compared with the control venous anastomoses (P < .05). CONCLUSIONS: These data demonstrate that local infusion of rFGF2-SAP significantly reduces venous anastomotic intimal hyperplasia and cell proliferation without systemic toxicity. This study suggests a new strategy for reducing intimal hyperplasia by the selective killing of proliferating smooth muscle cells with a potent chimeric mitotoxin through a novel local infusion device.

Biplexiform ganglion cells, characterized by dendrites in both outer and inner plexiform layers, are regular, mosaic-forming elements of teleost fish retinae.

Biplexiform ganglion cells were labelled by retrograde transport of HRP in five species of marine fish from the neoteleost acanthopterygian orders Perciformes and Scorpaeniformes. Their forms and spatial distributions were studied in retinal flatmounts and thick sections. Biplexiform ganglion cells possessed sparsely branched, often varicose, dendrites that ramified through the inner nuclear layer (INL) to reach the outer plexiform layer (OPL), as well as conventional arborizations in the most sclerad part of the inner plexiform layer (IPL). Their somata were of above-average size and were displaced into the vitread border of the INL. Mean soma areas ranged from 99 +/- 6 microns2 in Bathymaster derjugini (Perciformes) to 241 +/- 12 microns2 in Hexagrammos stelleri (Scorpaeniformes), but were similar in each species to those of the outer-stratified alpha-like ganglion cells, whose dendritic trees occupied the same IPL sublamina. In the best-labelled specimens, biplexiform cells formed clear mosaics with spacings and degrees of regularity much like those of other large ganglion cells, but spatially independent of them. Biplexiform mosaics were plotted in three species, and analyzed by nearest-neighbor distance and spatial correlogram methods. The exclusion radius, an estimate of minimum mosaic spacing, ranged from 113 microns in Hexagrammos stelleri, through 150 microns in Ernogrammus hexagrammus (Perciformes), to 240 microns in Myoxocephalus stelleri (Scorpaeniformes). A spatial cross-correlogram analysis of the distributions of biplexiform and outer-stratified alpha-like cells in Hexagrammos demonstrated the spatial independence of their mosaics. Similar cells were previously observed not only in the freshwater cichlid Oreochromis spilurus (Perciformes) but also in the goldfish Carassius auratus (Cypriniformes) which, being an ostariophysan teleost, is only distantly related. Thus, biplexiform ganglion cells may be regular elements of all teleost fish retinae. Their functional role remains unknown.

Local infusion of FGF-saporin reduces intimal hyperplasia.

The recent conjugation of the potent ribosome-inactivating protein saporin (SAP) with basic fibroblast growth factor (FGF2) to form recombinant (r)FGF2-SAP permits increased selectivity of this mitoxin for cells exhibiting upregulated FGF receptors. Systemic administration of rFGF-SAP in therapeutic doses, however, may be associated with significant liver toxicity. In this blinded study, we used a local boundary layer infusion approach to increase local drug concentration while minimizing the risk of side effects. Six dogs underwent bilateral carotid endarterectomies. Expanded polytetrafluoroethylene infusion devices, blindly primed with rFGF2-SAP to one artery or vehicle to the contralateral vessel, were anastomosed proximal to the injured segments so that each animal served as its own control. rFGF2-SAP (2 microgram/kg/day) or vehicle (5 microl/hr) was continuously delivered for 14 days from an osmotic reservoir, through the wall of the graft infusion device. Euthanasia was carried out at 14 days and the processed arteries were blindly analyzed for intimal thickening and cellular proliferation. All dogs survived until sacrifice with no clinical side effects. Liver function tests at euthanasia were not significantly altered when compared to baseline values. Intimal area in rFGF2-SAP-treated vessels averaged 0.31 +/- 0.10 mm2 versus 0.57 +/- 0.24 mm2 in the control segments (P = 0.02), a relative reduction of 46%. Cell proliferation, however, was not significantly different at 14 days postendarterectomy (2.40 +/- 1.31% vs 2.39 +/- 0.45%). From this study it can be concluded that locally delivered rFGF2-SAP reduces intimal hyperplasia and that the boundary layer infusion strategy is an effective means for delivering high local drug concentration while minimizing systemic drug effects.

Spatial properties of retinal mosaics: an empirical evaluation of some existing measures.

Mosaics of neurons are usually quantified by methods based on nearest-neighbor distance (NND). The commonest indicator of regularity has been the ratio of the mean NND to the standard deviation, here termed the 'conformity ratio.' However, an accurate baseline value of this ratio for bounded random samples has never been determined; nor was its sampling distribution known, making it impossible to test its significance. Instead, significance was assessed from goodness-of-fit to a Rayleigh distribution, or from another ratio, that of the observed mean NND to an expected mean predicted by theory, termed the dispersion index. Neither approach allows for boundary effects that are common in experimental mosaics. Equally common are 'missing' neurons, whose effects on the statistics have not been studied. To address these deficiencies, random patterns and real neuronal mosaics were analyzed statistically. Ns independent random-point samples of size Np were generated for 13 Np values between 25 and 6400, where Ns x Np > or = 144,000. Samples were generated with rectangular boundaries of aspect ratio 1:1, 1:5, and 1:10 to examine the influence of sample geometry. NND distributions, conformity ratios, and dispersion indices were computed for the resulting 45,997 independent random patterns. From these, empirical sampling distributions and critical values were determined. NND distributions for small-to-medium, bounded, random populations were shown to differ significantly from Rayleigh distributions. Thus, goodness-of-fit tests are invalid for most experimental mosaics. Charts are presented from which the significance of conformity ratios or dispersion indices can be read directly. The conformity ratio reacts conservatively to extremes of sample geometry, and provides a useful and safe test. The dispersion index is nonconservative, making its use problematic. Tests based on the theoretical distribution of the dispersion index are unreliable for all but the largest samples. Random deletions were also made from 33 real retinal ganglion cell mosaics. The mean NND, conformity ratio, and dispersion index were determined for each original mosaic and 36 independent samples at each of nine sampling levels, retaining between 90% and 10% of the original population. An exclusion radius, based on a spatial autocorrelogram, was also calculated for each of these 10,725 mosaic samples. The mean NND was moderately insensitive to undersampling, rising smoothly. The exclusion radius was remarkably insensitive. The conformity ratio and dispersion index fell steeply, sometimes failing to reach significance while half of the cells still remained. For the same 33 original mosaics, linear regression showed the exclusion radius to be 62 +/- 3% of the mean NND.

Large retinal ganglion cells in the channel catfish (Ictalurus punctatus): three types with distinct dendritic stratification patterns form similar but independent mosaics.

Retinal ganglion cells in the channel catfish (Ictalurus punctatus) were retrogradely labelled, and those with the largest somata and thickest primary dendrites were categorized by their levels of dendritic stratification. Three types were found, each forming a mosaic making up approximately 1% of the ganglion cell population. Using a system based on established sublaminar terminology, we call these the alpha-a (alpha a), alpha-b (alpha b), and alpha-c (alpha c) ganglion cell mosaics. Cells of the alpha a mosaic had large, sparsely branched trees in sublamina a at 10-30% of the depth of the inner plexiform layer (IPL), sclerad to those of all other large ganglion cells. Some alpha a somata were displaced into the IPL or inner nuclear layer (INL) but belonged to the same mosaic as their orthotopic counterparts. Cells of the alpha b mosaic had dendrites that branched a little more and arborized in sublamina b at 50-60% of the IPL depth. Many also sent fine branches into sublamina a, and some were fully bistratified in a and b. The alpha c cells arborized in the most vitread sublamina, sublamina c, at 80-95% of the IPL depth. The soma areas of the three types in the largest retina studied ranged between 139 microns 2 and 670 microns 2 with significant differences in the order alpha a > alpha c > or = alpha b. Analyses based on nearest-neighbour distance (NND) and on spatial auto- and cross-correlograms showed that each mosaic was statistically regular and independent of the others. Mosaic spacings were similar for each type, giving mean NNDs of 242-279 microns in the largest retina and 153-159 microns in a smaller one. Correspondences between these mosaics, previously defined large ganglion cell types in catfish, and other mosaic-forming large ganglion cells in fish and frogs are discussed along with their implications for neuronal classification, function, development, and evolution.