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Biomedical subjects

A Graham

Publications and source records attributed to A Graham.

At least 19 recordsLinked to original sources
PubMed2001

Synthesis of beta-galactose-conjugated chlorins derived by enyne metathesis as galectin-specific photosensitizers for photodynamic therapy.

A first report on the synthesis and biological evaluation of the beta-galactose-conjugated purpurinimides (a class of chlorins containing a six-membered fused imide ring system) as Gal-1 (galectin-1) recognized photosensitizers, prepared from purpurin-N-propargylimide via enyne metathesis, is discussed. On the basis of examination of the available crystal structure of the galectin-1 N-acetyllactose amine complex, it was considered that the chlorin-based photosensitizers could be introduced into a carbohydrate skeleton to expand the repertoire of the galectin-1-specific ligands. Preliminary molecular modeling analysis utilizing the modeled photosensitizers and the available crystal structures of galectin-carbohydrate complexes indicated that addition of the photosensitizer to the carbohydrate moiety at an appropriate position does not interfere with the galectin-carbohydrate recognition. Under similar drug and light doses, compared to the free purpurinimide analogue, the purpurinimides conjugated either with galactose or with lactose (Gal(beta1-4)-Glc) produced a considerable increase in photosensitizing efficacy in vitro. This indicates the possibility for development of a new class of specific photosensitizers for photodynamic therapy (PDT) based on recognition of a cellular receptor.

Carbohydrates↗
PubMed2001

The ectodermal placodes: a dysfunctional family.

The ectodermal placodes are focal thickenings of the cranial embryonic ectoderm that contribute extensively to the cranial sensory systems of the vertebrates. The ectodermal placodes have long been thought of as representing a coherent group, which share a developmental and evolutionary history. However, it is now becoming clear that there are substantial differences between the placodes with respect to their early development, their induction and their evolution. Indeed, it is now hard to consider the ectodermal placodes as a single entity. Rather, they fall into a number of distinct classes and it is within each of these that the members share a common development and evolution.

Animals↗
PubMed2001

Integration between the epibranchial placodes and the hindbrain.

Developmental integration results from coordination among components of different embryonic fields to realize the later anatomical and functional relationships. We demonstrate that in the chick head, integration between the epibranchial placodes and the hindbrain is achieved as the neuroglial hindbrain crest cells guide the epibranchial neuronal cells inward to establish their central connections. This work defines a role for the neuroglial hindbrain crest in organizing the afferent innervation of the hindbrain.

Animals↗
PubMed2001

A population of caudally migrating cranial neural crest cells: functional and evolutionary implications.

The deployment of the cranial neural crest is central to the patterning of the skeletomuscular elements of the vertebrate head, with cranial muscles invariably attaching to skeletal elements formed by crest from the same axial level. Here we demonstrate, through gene expression analysis, ablation studies and fate-mapping, the existence of a population of caudally migrating cranial crest that arise from the postotic neural tube. As with the rest of the postotic crest, these cells express the transcription factor Mafb, and this marker can be used to highlight their posterior migration. They pass out between the anterior somite and the otic vesicle, before turning caudally and running along the base of the somites. With long-term fate mapping, we show that these cells migrate to the clavicle and settle at the site of formation of the attachment point for the cleidohyoid muscle. As such, the influence of the cranial neural crest in organising skeletomuscular connectivity seems to extend beyond the head into the trunk. These results are of further importance as they help explain how, even though the pectoral girdle and the skull became physically dissociated during tetrapod evolution, skeletomuscular connectivity has been maintained.

Animals↗
PubMed2001

Triacylglycerol-rich lipoproteins alter the secretion, and the cholesterol-effluxing function, of apolipoprotein E-containing lipoprotein particles from human (THP-1) macrophages.

Elevated plasma levels of triacylglycerol-rich lipoproteins (TGRLP) are associated with increased risk of atherogenesis and abnormal reverse cholesterol transport, as illustrated in Type II diabetes. Here we examine the effect of plasma triacylglycerol-rich or cholesteryl ester-rich lipoproteins on the secretion of nascent apolipoprotein E (apoE)-containing lipoprotein E (LpE) particles by human (THP-1) macrophages. As expected, preincubation with low-density lipoprotein (LDL) yielded small but significant increases in total cellular cholesterol content and also the secretion of apoE by macrophages. By contrast, preincubation with TGRLP resulted in higher, dose-dependent, increases in apoE secretion that reflected, but were not dependent on, cellular triacylglycerol accumulation. Secreted apoE was incorporated into a pre-beta migrating LpE fraction that differed in lipid composition and flotation density depending on preincubation conditions. Specifically, the LpE-containing lipoprotein fraction produced by macrophages preincubated with TGRLP was cholesterol-poor, markedly heterogeneous and of higher peak flotation density (d 1.14-1.18) when compared with particles produced after preincubation with LDL. Both the conditioned medium and the isolated (d<1.21) LpE-containing fraction, yielded by macrophages preincubated with TGRLP, seemed poorer at inducing cholesterol efflux than the equivalent fractions from cells preincubated with LDL, as judged by [(3)H]cholesterol efflux from untreated 'naïve' macrophages. Thus, although the interaction of TGRLP with macrophages can enhance apoE output from these cells, the LpE particles produced seem to be relatively inefficient mediators of cholesterol efflux. These factors might contribute to the increased risk of atherosclerosis in individuals with Type II diabetes.

Apolipoproteins E↗
PubMed2001

A simple and efficient approach for the synthesis of fluorinated and nonfluorinated octaethylporphyrin-based benzochlorins with variable lipophilicity, their in vivo tumor uptake, and the preliminary in vitro photosensitizing efficacy.

Starting from commercially available Ni(II)octaethylporphyrin (OEP), an efficient approach for the preparation of a series of fluorinated and nonfluorinated benzochlorins with variable lipophicity has been developed. Their spectroscopic properties, preliminary in vitro photosensitizing efficacy, and tumor selectivity were determined. Our methodology provides a facile approach for the preparation of the free-base and the related Zn(II) benzochlorins containing alkyl and alkyl ether side chains with variable carbon units. For the preparation of benzochlorins containing alkyl groups attached to the exocyclic phenyl ring, the Ni(II) meso-(2-formylvinyl)octaethyl porphyrin 2 was reacted with various reagents such as (trifluoromethyl)trimethylsilane (TMS-CF3) or the Grignard reagents of various fluorinated or nonfluorinated alkyl halides. The corresponding intermediates 3, 6a-6e, and 8 obtained via intramolecular cyclization under acidic conditions afforded the related benzochlorins 5, 7a-d, and 9 in good yields except for 7e which was obtained in poor yield (11.4%). The alcohol 10 obtained by reacting porphyrin 2 with ethynylmagnesium chloride did not produce the expected acetylenic benzochlorin; instead the corresponding acetyl derivative 11 was obtained as a major product, which under appropriate reaction conditions was converted into a series of alkyl ether derivatives 13a-13d. To obtain a benzochlorin bearing an ester functionality (15), porphyrin 2 was first reacted with ethyl acetate/LDA and the intermediate alcohol 14 was then cyclized with sulfuric acid. Unlike most of the natural and synthetic chlorins, the Zn(II) complexes of the benzochlorin analogues exhibited a significant bathochromic shift ( approximately 10 nm) in the electronic absorption spectra, and the long wavelength absorptions were observed in the range 671-677 nm (epsilon: 43270-50360). For investigating the in vitro efficacy of these analogues, Molt-4 cells were used. At a concentration of 2.5 microM, and a light dose of 4 J/cm2, all benzochlorins produced significant photosensitizing efficacy. The tumor (RIF) and muscle uptake in C3H mice of these photosensitizers was determined by in vivo reflectance spectroscopy. These results indicate that in this series increasing the length of the alkyl or alkyl ether carbon chains at the fused phenyl ring system produced a significant increase in tumor uptake.

Animals↗
PubMed2001

Restricting Bmp-4 mediated apoptosis in hindbrain neural crest.

The segregation of the rhombencephalic neural crest into three streams, destined for particular pharyngeal arches, is a prominent feature of the developing vertebrate head and is likely necessary for normal morphogenesis. At least in part, the segregation of the crest into these streams involves the focal depletion of neural crest from rhombomeres 3 and 5 through large-scale apoptosis, mediated by the induction of Bmp-4 expression in the crest primordia of these two rhombomeres. Previously we found that in contrast to rhombomeres 3 and 5, the intervening segment rhombomere 4 was not susceptible to Bmp-4-sponsored neural crest cell death. To analyse the reason for this difference, we have isolated clones for the necessary components of the Bmp-4 signal transduction apparatus. We find that the hindbrain neural crest generally is primed to respond to Bmp-4 and that in vitro, besides rhombomere 3 and 5, rhombomeres 2 and 6 are also sensitive to Bmp-4-sponsored death. As before, however, we find that rhombomere 4 will not respond to this factor. Our analysis of the Bmp-4 antagonists has uncovered a reason for this. Rhombomere 4 expresses elevated levels of the antagonist Noggin at its dorsal midline just as crest production from this segment commences and, as development proceeds, in the crest that migrates away from there. At these later stages, expression also becomes apparent in the migratory post-otic crest as it flows by the otic vesicle. An interesting feature of these domains of Noggin expression is that they lie between territories of Bmp-4 expression, suggesting that Noggin is acting to protect from Bmp-4-mediated cell death.

Amino Acid Sequence↗
PubMed2001

Patterning the pharyngeal arches.

The presence of a muscularised pharynx with skeletal support is a fundamental vertebrate characteristic. Developmentally, the pharynx arises from the pharyngeal arches on either side of the head of vertebrate embryos. The development of the pharyngeal arches is complex involving a number of disparate embryonic populations, ectoderm, endoderm, neural crest and mesoderm, which must be co-ordinated to generate the components and overall identity of each of the arches. Previous studies suggested that it is the neural crest that plays a pivotal role in patterning the pharyngeal arches. It is now also becoming clear, however, that there are crest-independent patterning mechanisms. Therefore, pharyngeal arch patterning is more complex than was previously believed and there must be an integration of crest-dependent and -independent patterning mechanisms. BioEssays 23:54-61, 2001.

Animals↗
PubMed2001

Early development of the mesencephalic trigeminal nucleus.

The cells of the mesencephalic trigeminal nucleus (MTN) are the proprioceptive sensory neurons that innervate the jaw muscles. Interestingly, their evolution is generally thought to have been concomitant with that of the jaws. They are also the first born neurons of the mesencephalon, and their axons pioneer some of the major tracts within the brain. The cells of the MTN are also paradoxical in being the only group of intramedullary primary sensory neurons in amniotes. However, we know little about the early development of these important neurons, and we have analysed this here. To study the earliest stages of MTN development, we have used a battery of neural crest markers to try and pinpoint the progenitors of the MTN. We find that, contrary to current perceptions, the progenitors of the MTN are not highlighted by these markers, suggesting that they are not neural crest derived. However, the cells of the MTN are marked by means of their expression of Brn-3a. This gene labels cells that arise either side of the dorsal midline, extending rostrally from the isthmus across the roof of the mesencephalon. We have further demonstrated that the MTN develops under the influence of the Fgf-8 secreted by the isthmus. Ectopic Fgf-8 application promotes MTN development, whereas inhibiting Fgf-8 function in vivo drastically affects MTN development.

Animals↗
PubMed2001

Rhombomere interactions control the segmental differentiation of hindbrain neurons.

The embryonic hindbrain is subdivided into a series of metameric units termed rhombomeres, which display features that strongly suggest they are autonomous developmental units. However, some aspects of their phenotype develop nonautonomously. Here we have analyzed the possibility that interrhombomere interactions generate the pattern of segmental neuronal differentiation. The differentiation of both projection interneurons and motor neurons in the hindbrain is retarded in rhombomeres 3 and 5. We demonstrate here that if either rhombomere 3 or 5 is isolated from the influence of their neighbours, either in vitro or in vivo, then these segments no longer display delayed neuronal diufferentiation. We further show that the retardation of motor neurons differentiation in rhombomeres 3 and 5 is, at least in part, mediated by Bmp-4. If this molecule is inhibited, by grafting cells expressing chordin, then the motor neurons of these rhombomeres develop ahead of their normal schedule.

Activated-Leukocyte Cell Adhesion Molecule↗
PubMed2001

MRI of perineural extramedullary granulocytic sarcoma.

Granulocytic sarcoma is an extramedullary solid tumour consisting of myelogenous leukaemic blast cells, usually seen in acute myeloid leukaemia and less commonly in patients with chronic myeloid leukaemia or myeloproliferative disorders. Blast cells have a predilection for periosteal and perineural regions and rarely precede evidence of systemic disease. We present two patients, aleukaemic on peripheral blood counts, both at presentation and during subsequent treatment. We present the MRI features of this rare but important condition.

Adolescent↗
PubMed2001

Inability of plasma high-density lipoproteins to inhibit cell adhesion molecule expression in human coronary artery endothelial cells.

High-density lipoproteins (HDL) have several antiatherogenic actions, including the ability to sequester cellular cholesterol, to protect low-density lipoproteins from oxidation and to inhibit platelet aggregation. An early event in atherogenesis is the adhesion and recruitment of blood monocytes, a process mediated by cell adhesion molecules (CAMs), including vascular cell adhesion molecule-1 (VCAM-1) which is rapidly synthesized by endothelial cells in response to cytokines. It has been reported that HDL limits CAM expression in cultured human umbilical vein endothelial cells (HUVECs), implying that HDL also protects at an early stage in lesion development. Here, we have studied HDL suppression of CAM induction in human coronary artery endothelial cells (HCAECs), a model directly relevant to blood vessels susceptible to atherosclerosis. Arterial endothelial cells were preincubated with increasing amounts of total HDL, or different subfractions, and then activated with the inflammatory cytokine, tumor necrosis factor-alpha (TNF-alpha). Flow cytometric analysis failed to detect any downregulation of VCAM-1 or E-selectin expression by HDL in this model of vascular endothelium. Moreover, we were unable to confirm that HDL could suppress CAM induction in well-characterized, low-passage HUVECs, even though positive controls, 17beta-estradiol or a nitric oxide donor, did cause downregulation and factors such as variability in donors and HDL preparation, or culture conditions, were excluded. We tentatively conclude that, as isolated HDL did not downregulate CAM expression in cultured HCAECs or HUVECs, attenuation of CAM induction in arterial endothelium is unlikely to contribute to HDL antiatherogenic actions in vivo.

Arteries↗
PubMed2001

Stiles-Crawford effect of the first kind: assessment of photoreceptor alignments following dark patching.

Properties of presumed mechanisms controlling photoreceptor alignments are partially defined. A phototropic mechanism normally dominates alignment, but do modest changes in orientations occur with dark patching? Here, new photopic Stiles-Crawford (SCE-I) determinations were made before patching (pre-patch), just after 8-days of dark-patching (post-patch), and 3 days after patch removal (recovery test). We tested at 0, 11 and 22 degrees in the temporal retina of both eyes. Ten eyes of adult subjects were tested. SCE-I peak positions and Stile's parameter 'rho' were assessed. Dark-patching effects were small. Observations revealed meaningful corrective alignment overshoots with recovery in the light. Results suggest (1) the presence of multiple weak mechanisms affecting receptor alignments in the dark; (2) the phototropic mechanism to be dominant in the light; (3) the need for multiple test loci to be sampled in such studies, and (4) small changes in the SCE-I in the pupil plane can reflect meaningful events occurring at the retina.

Adaptation, Ocular↗
PubMed2001

Nuclear factor kappa B is involved in lipopolysaccharide-stimulated induction of interferon regulatory factor-1 and GAS/GAF DNA-binding in human umbilical vein endothelial cells.

1. In this study we examined the signalling events that regulate lipopolysaccharide (LPS)-stimulated induction of interferon regulatory factor (IRF)-1 in human umbilical vein endothelial cells (HUVECs). 2. LPS stimulated a time- and concentration-dependent increase in IRF-1 protein expression, an effect that was mimicked by the cytokine, tumour necrosis factor (TNF)-alpha. 3. LPS stimulated a rapid increase in nuclear factor kappa B (NFkappaB) DNA-binding activity. Pre-incubation with the NFkappaB pathway inhibitors, N-alpha-tosyl-L-lysine chloromethyl ketone (TLCK) or pyrrolidine dithiocarbamate (PDTC), or infection with adenovirus encoding IkappaBalpha, blocked both IRF-1 induction and NFkappaB DNA-binding activity. 4. LPS and TNFalpha also stimulated a rapid activation of gamma interferon activation site/gamma interferon activation factor (GAS/GAF) DNA-binding in HUVECs. Preincubation with the Janus kinase (JAK)-2 inhibitor, AG490 blocked LPS-stimulated IRF-1 induction but did not affect GAS/GAF DNA-binding. 5. Preincubation with TLCK, PDTC or infection with IkappaBalpha adenovirus abolished LPS-stimulated GAS/GAF DNA-binding. 6. Incubation of nuclear extracts with antibodies to RelA/p50 supershifted GAS/GAF DNA-binding demonstrating the involvement of NFkappaB isoforms in the formation of the GAS/GAF complex. 7. These studies show that NFkappaB plays an important role in the regulation of IRF-1 induction in HUVECs. This is in part due to the interaction of NFkappaB isoforms with the GAS/GAF complex either directly or via an intermediate protein.

Adenoviridae↗
PubMed2000

Development. Hear, hear, for the inner ear.

Although the development of the inner ear has been a favorite subject for biologists to study, it is not yet clear exactly which molecules are involved in the induction of the otic placode, the plug of embryonic ectoderm that will become the inner ear. In his Perspective, Graham takes us on an inner ear odyssey, explaining how the signaling molecules FGF-19 and Wnt-8c cooperate to induce formation of the otic placode (Ladher et al.).

Animals↗