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J J Walsh

Publications and source records attributed to J J Walsh.

At least 19 recordsLinked to original sources
PubMed2001

Induction of the mammalian node requires Arkadia function in the extraembryonic lineages.

The early mammalian embryo is patterned by signals emanating from extraembryonic and embryonic signalling centres, most notably the anterior visceral endoderm (AVE) and the node, respectively. The AVE is responsible for anterior development, whereas further axis specification depends on the node, the equivalent of Spemann's organizer. Formation of the node, at the anterior primitive streak, depends on expression of the transcription factor HNF3beta (ref. 4). However, both the source and the nature of the signals responsible for inducing the node have been unknown. Here we describe a recessive lethal mutation, arkadia, generated using gene-trap mutagenesis. Mutant embryos establish an AVE but fail to maintain anterior embryonic structures and lack a node. The mutation has disrupted the Arkadia gene, which encodes a putative intracellular protein containing a RING domain. Arkadia is essential for HNF3beta expression in the anterior primitive streak. Analysis with chimaeras, however, shows that Arkadia functions within extraembryonic tissues, revealing that these are required to induce the node. Furthermore, our experiments show that Arkadia interacts genetically with the transforming growth factor (TGF)beta-like factor Nodal, implying that Nodal mediates the function of Arkadia in node induction.

Amino Acid Sequence↗
PubMed2001

Arkadia enhances nodal-related signalling to induce mesendoderm.

Nodal-related members of the transforming growth factor (TGF)-beta family regulate the induction of mesoderm, endoderm, and mesendoderm, a tissue specific to the Spemann organizer. How these different tissues form in response to the same signalling molecules is not completely understood. It has been suggested that concentration-dependent effects, mediated by extracellular cofactors and antagonists, are responsible for the differences. Here we show that the nuclear protein Arkadia specifically potentiates the mesendoderm-inducing activity of a subset of TGF-beta family members. The combined activities of Arkadia and Xenopus nodal-related-1 are sufficient to induce mesendoderm and suppress mesoderm. Arkadia dorsalizes ventral tissues, resulting in the induction of organizer-specific gene expression. Blocking nodal signalling extracellularly inhibits these effects. Arkadia influences nodal activity when co-expressed and can function in cells adjacent to those producing the nodal signal. Our findings, together with the observation that Arkadia mutant mice lack a node and node-derived mesendoderm, identify Arkadia as an essential modulator of the nodal signalling cascade that leads to induction of Spemann's organizer.

Animals↗
PubMed2000

Diagnosis and management of hamate hook fractures.

Hamate hook fractures, although uncommon, are now recognized with increasing frequency because of the increase in popularity of racket sports and golf. Careful clinical evaluation and adjunctive radiologic investigation help establish the diagnosis. Acute nondisplaced fractures can heal with immobilization, while displaced fractures and nonunions typically require open reduction and internal fixation (acute fractures) or hook excision.

Athletic Injuries↗
PubMed1998

Binding of the alpha 2 integrin I domain to extracellular matrix ligands: structural and mechanistic differences between collagen and laminin binding.

The alpha 2 beta 1 integrin functions as a cell surface receptor for collagen on some cells and as both a collagen and laminin receptor on a more restricted subset of cell types including endothelial and epithelial cells. The alpha 2 integrin subunit I domain binds collagen in a divalent cation-dependent manner. In contrast, I domain binding to laminin occurs via both divalent cation-dependent and -independent mechanisms. Saturable binding was observed in the presence of either Mn2+ or EDTA, although the extent of binding in Mn2+ was twice that observed in EDTA. Half-maximal binding occurred at about 22 nM I domain in either case. Whereas laminin binding was significantly enhanced by Mn2+, with half-maximal binding occurring at 1.9 mM Mn2+, Mg2+ was much less effective. Deletion of the N-terminal 35 residues of the I domain, including the DXSXS portion of the MIDAS motif, caused a significant diminution of laminin binding activity. Laminin binding by the I domain was significantly inhibited by the alpha 2 beta 1 function-blocking antibody 6F1 in the presence of either EDTA or Mn2+. The non-function-blocking antibody 12F1 had no effect. In contrast to the binding of the alpha 2 integrin I domain to collagen, the laminin binding activity of the I domain was not enhanced by the addition of the first EF hand motif of the integrin.

Antibodies, Monoclonal↗
PubMed1997

Contributions of the I and EF hand domains to the divalent cation-dependent collagen binding activity of the alpha2beta1 integrin.

The alpha2beta1 integrin binds collagen in a Mg2+-dependent manner that is inhibited by Ca2+. Like the intact integrin, purified recombinant proteins containing the alpha2 integrin I domain, either alone or with variable numbers of alpha2 integrin EF hand metal binding sites, bound collagen in a Mg2+-dependent manner, and Ca2+ did not support binding. However, unlike the intact integrin, Ca2+ did not inhibit the Mg2+-dependent binding of any of the fusion proteins to collagen. Binding to collagen was saturable and blocked by the alpha2beta1 function blocking antibody 6F1. Deletional analysis demonstrated that residues present within the amino-terminal 35 amino acids contribute to the 6F1 epitope and are required for Mg2+-dependent collagen binding. The results indicate that the I domain contains a Mg2+ binding site that is essential for collagen binding and that the I domain alone is sufficient for collagen binding. Binding is markedly enhanced in a divalent cation-dependent manner by the addition of the first EF hand motif. Mutation of the EF hand to an inactive form completely abrogated the effect. The sites necessary for Ca2+ inhibition are not present within the I domain or the adjacent region containing the three EF hand sites.

Binding Sites↗
PubMed1997

The alpha 2 beta 1 integrin binds to the CB4 peptide of the alpha 2(I) collagen chain.

The adhesion of platelets and other cells to type I collagen is mediated by the alpha 2 beta 1 integrin. A binding site for the alpha 2 beta 1 integrin within the alpha 1(I) collagen chain has previously been localized to the cyanogen bromide fragment alpha 1(I)-CB3. We noe show by use of inhibitory monoclonal antibodies against the alpha 2 beta 1 integrin, that platelets also adhere to purified alpha 2(I) collagen chains by a mechanism mediated by the alpha 2 beta 1 integrin. Moreover, following isolation of cyanogen bromide fragments of the alpha 2(I) collagen chain by HPLC, we demonstrate that alpha 2 beta 1 integrin-mediated adhesion is restricted to the CB4 fragment of the alpha 2(I) collagen polypeptide. These findings indicate the presence of at least two spatially distinct binding sites for the alpha 2 beta 1 integrin on the native type I collagen triple helix.

Animals↗
PubMed1993

Platelet interactions with fibronectin: divalent cation-independent platelet adhesion to the gelatin-binding domain of fibronectin.

Divalent cation-dependent platelet adhesion to fibronectin (FN) is mediated by the integrin receptors alpha 5 beta 1 (GP Ic-IIa) and alpha IIb beta 3 (GP IIb-IIIa), which recognize the RGD (Arg-Gly-Asp) sequence in the cell-binding domain. However, FN can also support divalent cation-independent platelet adhesion. To determine which domain of FN mediates divalent cation-independent adhesion, proteolysis with thermolysin and affinity chromatography were used to isolate the cell-binding, gelatin-binding, and heparin-binding domains of FN. Unactivated and thrombin-activated platelets adhered to intact FN and the 45-Kd gelatin-binding domain in the presence of either Ca2+ or EDTA. Platelet spreading was mediated only by the 105-Kd cell-binding domain and required divalent cations. The heparin-binding domains did not support platelet adhesion. Reduction of intrachain disulfide bonds or removal of carbohydrate side chains on the gelatin-binding domain did not alter the ability to support platelet adhesion. Divalent cation-independent adhesion to the 45-Kd gelatin-binding domain was not inhibited by RGDS (Arg-Gly-Asp-Ser) synthetic peptides or monoclonal antibodies (MoAbs) directed against known platelet receptors. We conclude that platelets can adhere but not spread on the gelatin-binding domain of FN by a novel divalent cation-independent mechanism.

Amino Acid Sequence↗
PubMed1993

Auto-induction of transforming growth factor-beta in human lung fibroblasts.

The type beta transforming growth factors (TGF-beta s) are a family of potent cytokines with diverse effects on proliferation, differentiation, turnover of extracellular matrix components, oncogene expression, and other aspects of cellular phenotype. Unlike lung fibroblasts of certain species, unstimulated human lung fibroblast lines produce little or no TGF-beta in culture. However, TGF-beta has been reported to autoregulate its own production in certain human tumor cells and in rodent cell lines. To test whether this phenomenon is operative in fibroblasts from normal human lung tissue, confluent cultures of IMR90 normal fetal lung fibroblasts were exposed to TGF-beta. Cultures were exposed briefly to purified TGF-beta 1 under serum-free conditions and secretion of newly synthesized TGF-beta over the ensuing 72 h was determined by immunoblotting and bioassays made specific with the use of neutralizing antibodies. Steady-state levels of mRNA for TGF-beta 1 were detected by Northern and slot blot hybridization analysis of total cellular RNA. The 2.5 kb TGF-beta 1 mRNA species rose within 1.5 h of exposure of IMR90 cells to TGF-beta 1 and reached maximal levels after 16 h. Increased levels of TGF-beta were detected in conditioned medium 9 h after the start of the exposure. Thereafter, TGF-beta continued to accumulate at an elevated rate (90 +/- 7 versus < or = 15 pg/10(6) cells/h in uninduced cells) for up to 72 h. As little as 1 ng/ml TGF-beta 1 auto-induced TGF-beta secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Distinct determinants on collagen support alpha 2 beta 1 integrin-mediated platelet adhesion and platelet activation.

Recent studies have revealed that the sequence of amino acids asp-gly-glu-ala represents an essential determinant of the site within the alpha 1(I)-CB3 fragment of collagen recognized by the alpha 2 beta 1 integrin cell surface collagen receptor (Staatz et al., 1991). Studies employing chemical modifications of collagen amino acid side chains confirm both the essential nature of the acidic side chains of aspartic acid and glutamic acid residues and the nonessentiality of lysine epsilon-amino groups in supporting adhesion mediated by the alpha 2 beta 1 integrin. The approach also indicates the presence of a distinct determinant on collagen separate from the alpha 2 beta 1 recognition site that contains essential lysine side chains and that is necessary for subsequent interactions with the platelet surface that give rise to collagen-induced platelet activation and secretion. The two-step, two-site model for cellular signaling involving both an integrin and a signal-transducing coreceptor suggested by these data may be common to other integrin-mediated processes.

Acetylation↗
PubMed1991

Plasma cell pneumonia induced by Legionella pneumophila.

We report a case of severe Legionella pneumonia in which the histopathologic features of an open lung biopsy specimen were characterized by a monotypic plasma cell infiltrate and absence of granulocytes or other acute inflammatory cells. The clinical course was otherwise typical for a severe case of Legionella pneumonia with complete functional and roentgenographic recovery following a prolonged convalescence. We speculate on possible reasons for this patient's unusual histopathologic features.

Female↗
PubMed1990

The alpha 2 beta 1 integrin cell surface collagen receptor binds to the alpha 1 (I)-CB3 peptide of collagen.

We have previously shown that platelets adhere to collagen substrates via a Mg2(+)-dependent mechanism mediated by the surface glycoprotein Ia-IIa (human leukocyte very late activation protein 2, alpha 2 beta 1 integrin) complex. The adhesion is specific for collagen and is supported by collagen types I, II, III, IV, and VI. Several other members of the integrin family of adhesive protein receptors recognize discrete linear amino acid sequences within their adhesive glycoprotein ligands. Experiments with both intact platelets and with liposomes containing the purified receptor complex indicated that the alpha 2 beta 1 receptor recognized denatured type I collagen in a Mg2(+)-dependent manner. To further localize the binding site, the alpha 1 and alpha 2 chains of type I collagen were purified by gel filtration and ion exchange chromatography and tested as adhesive substrates. Both the alpha 1(I) and alpha 2(I) chains effectively supported Mg2(+)-dependent platelet adhesion. The purified alpha 1(I) collagen chain was then subjected to cleavage with cyanogen bromide, and the resultant peptides were separated by chromatography on carboxymethylcellulose. Only the alpha 1(I)-CB3 fragment supported Mg2(+)-dependent platelet adhesion. The monoclonal antibody P1H5 which recognizes an epitope on the alpha 2 subunit of the integrin receptor and which inhibits the adhesion of both intact platelets and liposomes bearing the purified receptor to collagen also inhibited platelet adhesion to the alpha 1(I)-CB3 fragment. These results indicate that the alpha 2 beta 1 receptor recognizes a sequence of amino acids present in the alpha 1(I)-CB3 fragment of type I collagen. An identical or similar sequence likely mediates binding of the receptor to other collagen polypeptides.

Antibodies, Monoclonal↗
PubMed1989

Freedom Set II--a Y-set CAPD system with a novel design.

The Freedom Set II is designed with a three-way valve to control the direction of dialysate flow thus simplifying the procedure and decreasing training time. Twenty-four patients from two centers were trained on the Freedom Set II and were observed for ninety patient months. Five patients who had been on other bagless systems expressed their preference for the Freedom Set II. The peritonitis rate for the observation period was one episode every 21 patient months. We conclude that the Freedom Set II provides safe, cost-effective dialysis and simplifies the CAPD procedure.

Consumer Behavior↗
PubMed1988

Long-term results of infragenicular bypasses with autogenous vein originating from the distal superficial femoral and popliteal arteries.

Forty-nine bypasses originating from the distal superficial femoral artery or popliteal artery in 46 patients were reviewed to examine late patency, limb salvage, and factors leading to graft failure. Operations were performed because of tissue loss in 86%, rest pain in 12%, and limiting claudication in 2% of limbs. Proximal anastomosis was from the distal superficial femoral artery in 12% and the popliteal artery in 88%. Distal anastomosis was to the below-knee popliteal artery or proximal tibial vessels in 20% and the distal tibial vessels in 80%. Life-table analysis showed a primary patency rate of 83%, 62%, and 41%, at 1, 3, and 5 years, respectively. The rate of limb salvage at 6 years for all grafts was 69%. Cox proportional hazards analysis determined that stenosis of 20% or greater in the proximal superficial femoral artery before bypass was a significant risk factor for graft failure (p = 0.02) despite the presence of normal intra-arterial pressure at the site of the proximal anastomosis at the time of bypass. Long-term survival in these patients was low, with a 6-year survival rate of only 24%. Infragenicular bypasses originating from the distal superficial femoral artery or the popliteal artery can be performed with patency and limb salvage rates comparable to bypasses originating from the common femoral artery. These bypasses are more likely to fail when performed in the presence of a stenosis 20% or greater in the superficial femoral or popliteal artery proximal to the graft origin.

Aged↗
PubMed1988

Vein compression by arterial aneurysms.

This study was done to emphasize the importance of early, accurate diagnosis of arterial aneurysms that show the symptoms of venous obstruction. Fourteen patients were identified as having atherosclerotic aneurysms producing venous compression. Nine patients had popliteal aneurysms, causing popliteal vein thrombosis in three patients and vein compression without thrombosis in six patients. Five patients had iliac artery aneurysms, producing left iliac vein thrombosis in one patient and venous compression without thrombosis in four patients. In 10 patients the cause of the venous compression symptoms was correctly identified and appropriate revascularization was performed with successful results. In four patients, two with iliac artery aneurysms and two with popliteal artery aneurysms, the associated aneurysm was not identified. One patient died of a ruptured aneurysm and three patients had below-knee amputations because of untreatable distal ischemia. Inappropriate treatment of patients with venous obstruction from unrecognized arterial aneurysms is associated with unacceptable morbidity and mortality. Accurate diagnosis with timely aneurysm repair eliminates the risk of aneurysm rupture or thrombosis and simultaneously alleviates venous compression symptoms.

Aged↗
PubMed1982

Blood levels of caffeine and results of fetal examination after oral administration of caffeine to pregnant rats.

Pregnant FDA-strain Osborne-Mendel rats were administered repeated doses of caffeine by oral intubation (gavage) and by administration in the drinking water (ad libitum sipping). When [1-methyl-14C]caffeine was administered at a dosage of 80 mg per kg per day by ad libitum sipping on days 12 to 15 of gestation, the amounts of radioactivity in blood were variable; the highest level on day 12 was 0.2% of the dose per ml of blood. The highest blood level of caffeine observed during a 24-h sampling period averaged 5.7 micrograms ml-1. When [14C]caffeine was administered by gavage at a dosage of 80 mg kg-1 on day 12, the blood level of radioactivity reached a peak of 0.4% of the dose per ml of blood and declined rapidly thereafter. The highest amount of caffeine observed in blood averaged 63.1 micrograms ml-1, 1 h after gavage. The overall blood elimination half-life of radioactivity in pregnant rats treated by gavage was 2.6 h, and the half-life of caffeine in blood was 1.7 h. The levels of radioactivity in the fetus and maternal muscle per unit weight were comparable after each method of administration. A comparison of autopsy results from both groups indicated that resorptions were increased when compared with rats that did not receive caffeine; this effect was more marked in the gavage group than in the ad libitum sipping group. Ectrodactyly was observed only in offspring of the gavage group. The incidences of ectrodactyly or resorptions did not appear to be directly related to nutrition or fluid intake.

Administration, Oral↗