Ultrasonography of the medial palmar intercarpal ligament in the Thoroughbred: technique and normal appearance.

Reasons For Performing Study: Tearing of the medial palmar intercarpal ligament (MPICL) has been recognised as a cause of lameness in the Thoroughbred, but diagnosis is difficult due to the nonspecific clinical signs, and can be achieved only by performing arthroscopy on the mid carpal joint (MCJ). It would be beneficial to be able to image the MPICL using ultrasonography to determine whether pathology is present in the ligament in order to aid diagnosis and prognosis.

Objectives: To determine whether the MPICL could be imaged using ultrasound from the dorsal aspect of the MCJ, and to describe the technique and normal ultrasonographic appearance of the ligament.

Inducible short-term and stable long-term cell culture systems reveal that the PAX3-FKHR fusion oncoprotein regulates CXCR4, PAX3, and PAX7 expression.

In the pediatric cancer alveolar rhabdomyosarcoma (ARMS), the 2;13 chromosomal translocation juxtaposes the PAX3 and FKHR genes to generate a chimeric transcription factor. To explore molecular pathways altered by this oncoprotein, we generated an inducible form by fusing PAX3-FKHR to a modified estrogen receptor ligand-binding domain and expressed this construct in the RD embryonal rhabdomyosarcoma cell line. This inducible system permits short-term evaluation of downstream expression targets of PAX3-FKHR and complements a panel of stable long-term RD subclones constitutively expressing PAX3-FKHR.

Alpine pathways of membrane traffic.

Workshop on Protein Sorting in the Secretory Pathway

I-proteins - a proposed switch in myotubularin function.

In this article, we consider the functions of the myotubularins - a large family of phosphoinositide 3-phosphatases. By analogy with the phosphatidylinositol 3-phosphatase PTEN (phosphatase and tensin homolog deleted in chromosome ten) and many protein phosphatases, it has been proposed that the primary function of this protein family is to regulate substrate levels, in this case phosphatidylinositol (3)-phosphate. We propose an alternate, or additional, function that is analogous to the G-protein family of phosphatases, which use nucleotide-dependent conformational changes to transduce signals or do mechanical work.

Radiographic progression of osteoarthritis of the canine stifle joint: a prospective study.

A radiographic scoring system for features of osteoarthritis of the canine stifle joint was devised. Intra- and inter-rater agreement was assessed by calculation of unweighted kappa statistics. "Global score," "effusion," "osteophytosis," and "intra-articular mineralization" were all features that had acceptable reliability, but "subchondral sclerosis" did not.

Golgi inheritance: shaken but not stirred.

Our view of what happens to the Golgi and ER during mitosis in mammalian cells has been shaken once more. Rather than the Golgi contents being recycled through, or mixed with the ER, two recent studies taking complementary approaches, find that the contents of these organelles remain separate throughout mitosis.

Environmentally determined genetic expression: clinical correlates with molecular variants of carbamyl phosphate synthetase I.

Carbamyl phosphate synthetase I (CPSI) determines the rate-limiting entry of free ammonia into the urea cycle. Disruption of CPSI affects the liver's ability to remove waste nitrogen and produce arginine, citrulline, and urea. Arginine is the necessary precursor for the critical biomolecule, nitric oxide (NO).

Thoracic high-resolution computed tomography in the diagnosis of metastatic carcinoma.

Three dogs were presented for investigation of recurrent pyrexia of unknown origin, chronic vomiting and respiratory distress, respectively. One dog was markedly underweight and the other two were cachexic. Physical examination and initial diagnostic tests failed to establish the underlying cause of the presenting signs.

Dynamics of Golgi matrix proteins after the blockage of ER to Golgi transport.

When the ER to Golgi transport is blocked by a GTP-restricted mutant of Sar1p (H79G) in NRK-52E cells, most Golgi resident proteins are transported back into the ER. In contrast, the cis-Golgi matrix proteins GM130 and GRASP65 are retained in punctate cytoplasmic structures, namely Golgi remnants. Significant amounts of the medial-Golgi matrix proteins golgin-45, GRASP55 and giantin are retained in the Golgi remnants, but a fraction of these proteins relocates to the ER.

YSK1 is activated by the Golgi matrix protein GM130 and plays a role in cell migration through its substrate 14-3-3zeta.

The Golgi apparatus has long been suggested to be important for directing secretion to specific sites on the plasma membrane in response to extracellular signaling events. However, the mechanisms by which signaling events are coordinated with Golgi apparatus function remain poorly understood. Here, we identify a scaffolding function for the Golgi matrix protein GM130 that sheds light on how such signaling events may be regulated.

Membrane fusion: caught in a trap.

SNAREs are small coiled-coil proteins required for specific membrane fusion events in eukaryotic cells. Recent evidence points to the existence of an inhibitory class of SNAREs, i-SNAREs, which prevent incorrect fusions from occurring, adding a further layer of regulation to the process of membrane docking and fusion.

Sex hormone patterns and serum retinol concentrations in adolescent girls.

Objective: To describe the pattern and relation of sex serum hormones and retinol concentrations over 1 menstrual cycle in adolescent girls living in a resource-poor setting.

Study Design: Venous blood samples were collected on alternate days of the cycle, and estrogen, progesterone, luteinizing hormone (LH), follicle stimulating hormone and serum retinol were measured. A linear random effects model was used to examine the relationship between sex hormones and serum retinol.

Computerized physician order entry and medication errors in a pediatric critical care unit.

Objective: Medication errors are a major concern of health care professionals and medical institutions, especially errors involving children. Studies in adults have shown that computerized physician order entry (CPOE) systems reduce medication errors and adverse drug events (ADEs). The effect of CPOE implementation in a pediatric population has not been reported.

Fatal pulmonary hypertension associated with pertussis in infants: does extracorporeal membrane oxygenation have a role?

Objective: The deaths of 4 infants who had confirmed pertussis infection at our hospital during the past year motivated us to review their cases as well as the use of extracorporeal membrane oxygenation (ECMO) in respiratory failure caused by pertussis.

Methods: Retrospective chart reviews of the 4 infants who had pertussis and died at Vanderbilt University Medical Center (VUMC) from May 2001 to May 2002 were conducted. The Extracorporeal Life Support Organization (ELSO) database is an international voluntary registry established in 1986 to compile passive reports of adult and pediatric patients placed on ECMO.

The UIM domain of Hrs couples receptor sorting to vesicle formation.

Hepatocyte growth factor regulated tyrosine kinase substrate (Hrs), a main component of the 'bilayered' clathrin coat on sorting endosomes, was originally identified as a substrate of activated tyrosine kinase receptors. We have analysed Hrs phosphorylation in response to epidermal growth factor (EGF) stimulation and show that the evolutionary conserved tyrosines Y329 and Y334 provide the principal phosphorylation sites. Hrs is proposed to concentrate ubiquitinated receptors within clathrin-coated regions via direct interaction with its UIM (ubiquitin interaction motif) domain.

Phosphorylation of mitotic kinesin-like protein 2 by polo-like kinase 1 is required for cytokinesis.

We have investigated the function of mitotic kinesin-like protein (MKlp) 2, a kinesin localized to the central spindle, and demonstrate that its depletion results in a failure of cleavage furrow ingression and cytokinesis, and disrupts localization of polo-like kinase 1 (Plk1). MKlp2 is a target for Plk1, and phosphorylated MKlp2 binds to the polo box domain of Plk1. Plk1 also binds directly to microtubules and targets to the central spindle via its polo box domain, and this interaction controls the activity of Plk1 toward MKlp2.

Golgins in the structure and dynamics of the Golgi apparatus.

Golgins are a family of coiled-coil proteins associated with the Golgi apparatus necessary for tethering events in membrane fusion and as structural supports for Golgi cisternae. Recent work has shown that golgins such as GM130, golgin-45 and p115 bind to Rab GTPases via their coiled-coil domains, and that GM130, rather than being part of a static structural matrix, is in dynamic exchange between the membrane surface and the cytoplasm. Golgins such as bicaudal-D1 and -D2 bind to Rab6, but, rather than tethering membranes together, link vesicles to the cytoskeleton, thus adding a new function for this class of proteins.

Arfophilins are dual Arf/Rab 11 binding proteins that regulate recycling endosome distribution and are related to Drosophila nuclear fallout.

Arfophilin is an ADP ribosylation factor (Arf) binding protein of unknown function. It is identical to the Rab11 binding protein eferin/Rab11-FIP3, and we show it binds both Arf5 and Rab11. We describe a related protein, arfophilin-2, that interacts with Arf5 in a nucleotide-dependent manner, but not Arf1, 4, or 6 and also binds Rab11.

Membrane traffic: a glitch in the Golgi matrix.

Golgins are coiled-coil proteins thought to form a matrix important for shaping and organising Golgi cisternae and directing long-range recognition events in vesicular transport. This model is brought into question by new evidence that two golgins, GM130 and golgin-84, contribute to but are not essential for protein transport and Golgi structure.

Phosphatidylinositol-5-phosphate activation and conserved substrate specificity of the myotubularin phosphatidylinositol 3-phosphatases.

Phosphoinositides control many different processes required for normal cellular function. Myotubularins are a family of Phosphatidylinositol 3-phosphate (PtdIns3P) phosphatases identified by the positional cloning of the MTM1 gene in patients suffering from X-linked myotubular myopathy and the MTMR2 gene in patients suffering from the demyelinating neuropathy Charcot-Marie-Tooth disease type 4B. MTM1 is a phosphatidylinositol phosphatase with reported specificity toward PtdIns3P, while the related proteins MTMR2 and MTMR3 hydrolyze both PtdIns3P and PtdIns(3,5)P2.

Immunophenotype of desmoplastic small round cell tumors as detected in cases with EWS-WT1 gene fusion product.

Desmoplastic small round cell tumor is a rare tumor typically involving peritoneum. Although the histogenesis of desmoplastic small round cell tumor has yet to be elucidated, immunophenotypical and morphological analysis shows a characteristic divergent phenotype overlapping with other round cell tumors such as Ewing's sarcoma/primitive neuroectodermal tumor, rhabdomyosarcoma, small cell mesothelioma, and carcinoma. Detection of the EWS-WT1 gene fusion is characteristic of desmoplastic small round cell tumor and has been used reliably in tumor diagnosis.

Effect of cardiopulmonary bypass on urea cycle intermediates and nitric oxide levels after congenital heart surgery.

Objective: To test the hypothesis that cardiopulmonary bypass used for repair of ventricular septal defects and atrioventricular septal defects would decrease availability of urea cycle intermediates including arginine and subsequent nitric oxide availability.

Study Design: Consecutive infants (n = 26) undergoing cardiopulmonary bypass for repair of an unrestrictive ventricular septal defect or atrioventricular septal defect were studied. Blood samples were collected immediately before surgery, immediately after surgery, and 12 hours, 24 hours, and 48 hours after surgery.

Ethnic diversity in a critical gene responsible for glutathione synthesis.

The tripeptide glutathione is an important biomolecule that acts as a scavenger of free radicals and plays a role in a number of other cellular processes. A number of diseases, including Parkinson's disease, cancer, sickle cell anemia, and HIV infection, are thought to involve oxidative stress and depletion of glutathione. The heterodimeric enzyme glutamate cysteine ligase catalyzes the first, rate-limiting step in the de novo synthesis of glutathione.

Vesicular transport.

The minimal requirements for COP I and COP II vesicle formation from liposomes are the small GTPases Arf1 and Sar1, GTP and the coat complexes themselves. Coat-nucleating factors including specific GTP-GDP exchange factors, SNAREs and other cargo molecules are likely to determine the site of vesicle formation. Tethering mediated by long rod-like coiled-coil proteins precedes docking of vesicles with their target membrane and contributes to specific recognition of this membrane.