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Targeted mutation of Cyln2 in the Williams syndrome critical region links CLIP-115 haploinsufficiency to neurodevelopmental abnormalities in mice.

Williams syndrome is a neurodevelopmental disorder caused by the hemizygous deletion of 1.6 Mb on human chromosome 7q11.23. This region comprises the gene CYLN2, encoding CLIP-115, a microtubule-binding protein of 115 kD. Using a gene-targeting approach, we provide evidence that mice with haploinsufficiency for Cyln2 have features reminiscent of Williams syndrome, including mild growth deficiency, brain abnormalities, hippocampal dysfunction and particular deficits in motor coordination. Absence of CLIP-115 also leads to increased levels of CLIP-170 (a closely related cytoplasmic linker protein) and dynactin at the tips of growing microtubules. This protein redistribution may affect dynein motor regulation and, together with the loss of CLIP-115-specific functions, underlie neurological alterations in Williams syndrome.

Animals↗

The mechanism of renomedullary antihypertensive action: haemodynamic studies in hydronephrotic rats with one-kidney renal-clip hypertension.

1. The protective action of the renal medulla was studied in one-kidney renal-clip hypertension in rats with unilateral hereditary hydronephrosis and almost complete atrophy of the medulla of the affected kidney. 2. Rats were unilaterally nephrectomized. The first group had a normal kidney remaining, and the animals from the second and third groups were left with a hydronephrotic kidney and received renomedullary and renocortical autotransplants respectively. Two weeks later all rats were made hypertensive by placing a silver clip (0.2 mm) on the renal artery. 3. From the fourth day after clipping until the end of the experiment blood pressure was found to be significantly (P less than 0.01) lower in rats with medullary transplants than in the other groups. No differences in renal excretory function, plasma volume and plasma renin activity were found between the groups either before or during development of hypertension (5 and 21 days after clipping). Early in the course of hypertension (5 days) cardiac output was significantly (P less than 0.05) lower in the rats with medullary transplants than in the other groups, although an increase in plasma volume was noted in all three groups. At that time no difference in total peripheral resistance was found between the groups. 4. The results are consistent with the hypothesis that the renomedullary antihypertensive substance(s) mitigates hypertension by preventing a hypertensive haemodynamic response to sodium/volume overload.

Animals↗

Retention and postinsertion maintenance of bar-clip, ball and magnet attachments in mandibular implant overdenture treatment: an in vivo comparison after 3 months of function.

It could be hypothesised that attachments, which provide more retention against vertical and horizontal dislodgement, will be associated with more favourable parameters of oral function. This in vivo study is designed to provide data regarding initial retention force, loss of retention force after 3 months of function and postinsertion maintenance and complications associated with the use of magnet, bar-clip and ball attachments in mandibular overdenture treatment. Eighteen edentulous subjects received two permucosal implants in the inter-foramina region of the mandible, a new denture and three successive suprastructure modalities (magnet-, bar-clip and ball attachments). The retention force of the attachments at baseline and after 3 months was measured in a standardised way. The amount and type of postinsertion maintenance that was related to the attachment were evaluated. No differences in retention force at baseline and after 3 months of loading were observed for all three attachment types. The mean retention forces of magnet attachments, bar-clip attachments and ball attachments were 8.1, 31.3 and 29.7 N respectively. Functional maintenance complications related to the attachments were predominantly observed in 11/36 magnet attachments. Functional problems in the ball attachment group were relatively rare, easily manageable and seen in 4/36 attachments. The bar-clip attachments exhibited no maintenance problems at all.

Adult↗

Use of endoscopic mucosal clips in radiotherapy planning for oesophageal carcinoma: a series of three cases.

This paper describes the technique of placing endoscopic mucosal clips to localize oesophageal carcinoma and hence facilitate radiotherapy planning. This technique has been used on three patients in our centre. One was treated radically with external beam radiotherapy and two were treated palliatively (retreatment) with intraluminal brachytherapy. Mucosal hemoclips were placed at the time of endoscopy to indicate the superior and inferior extent of the tumour. The clips provided a radiologically-recognisable marker of the tumour extent and were visible on simulation films or planning CT scans. The radiation portal included the tumour as demarcated by the clips with an adequate margin. There were no complications related to the placement of the clips. All patients completed the radiotherapy course as planned.

Aged↗

Chronic anti-Thy-1 nephritis is aggravated in the nonclipped but not in the clipped kidney of Goldblatt hypertensive rats.

BACKGROUND: We have previously shown that renovascular hypertension does not inhibit healing of the acute Thy-1 nephritis. To test whether a chronic model of the Thy-1 nephritis is more susceptible to high blood pressure, the repetitive hit model was evaluated in rats with 2-kidney, 1-clip Goldblatt hypertension. METHODS: Six weeks after initiation of 2-kidney, 1-clip hypertension, chronic Thy-1 glomerulonephritis was induced in hypertensive rats by four consecutive injections of rabbit antiserum in weekly intervals. Renal structure and function were examined two weeks after the last injection. Glomerular binding of rabbit IgG as well as expression of transforming growth factor-beta (TGF-beta), alpha-smooth muscle actin (alpha-SMA) and cyclooxygenase (COX)-1 and -2 were evaluated by Western blotting. RESULTS: Similar glomerular deposition of rabbit IgG was detected in normotensive rats and in both kidneys of Goldblatt hypertensive rats indicating similar delivery and binding of the heterologous antibody. Induction of the repetitive Thy-1 model significantly enhanced glomerular damage in the nonclipped kidney and increased albuminuria. Surprisingly, no glomerular damage developed in the clipped kidney of nephritic hypertensive rats. In contrast, increased glomerular volume and increased expression of TGF-beta, alpha-SMA as well as COX-1 and COX-2 were found in normotensive nephritic rats and in both kidneys of nephritic hypertensive rats. CONCLUSION: Glomerular and tubulointerstitial damage of the chronic Thy-1 model is dramatically enhanced in the nonclipped kidneys of Goldblatt hypertensive rats. In contrast, the clipped kidney is completely protected from this immunological injury despite similar activation of glomerular cells, induction of TGF-beta, COX-1 and COX-2 and glomerular hypertrophy.

Albuminuria↗

Non-penetrating vascular clips anastomosis inhibited intimal thickening under poor runoff conditions in canine autogenous vein grafts.

OBJECTIVE: Late graft failure is still a significant problem, particularly in cases with poor runoff vessels. The main cause of late graft failure is intimal thickening of the anastomotic region. Vascular closure system (VCS) clips may provide ideal anastomosis, since they do not penetrate the wall. Therefore, we examined whether the VCS clips affect intimal thickening under poor runoff conditions in the canine autogenous vein grafts. METHODS: A canine poor runoff model was prepared at both femoral veins. Four weeks after the first surgical procedure, two groups were established according to the two different methods of anastomosis employed. The right femoral vein graft was performed using polypropylene sutures, conventional surgical anastomosis (control group), while the left femoral vein graft was performed using VCS clips anastomosis (VCS group). Four weeks after grafting, the vein grafts were removed and the intimal thickening of proximal, distal anastomosis and midportion of the vein grafts were examined histologically. RESULTS: In the control group, flow rate and variation were 26+/-8 ml/min and 51+/-10 dynes/cm(2), respectively. In the VCS group, the flow rate and variation were 23+/-11 ml/min and 44+/-14 dynes/cm(2), respectively. There were no significant differences between the two groups. The average value of intimal thickening of both the anastomotic region and the midportion of the vein graft in the VCS group was significantly inhibited compared to that of the control group. The number of positive cells of masson trichrome stain in the VCS group was significantly less than that of the control group. CONCLUSIONS: These experiments indicate that VCS clips significantly inhibit intimal thickening under poor runoff conditions in canine autogenous vein grafts to a greater extent compared to suture-constructed anastomosis. One mechanism that may account for the decreased intimal thickening is the inhibition of the expression of transforming growth factor-beta (TGF-beta), because the number of positive cells of masson trichrome stain in the VCS group was significantly less than that of the control group.

Animals↗

An inexpensive self fabricated pressure clip for the ear lobe.

Pressure therapy is an integral part of keloid treatment. There are certain areas of the body which are prone to keloid formation but are not amenable to pressure therapy. The ear lobe is one such area. A simple, self fabricated and inexpensive pressure clip has been used for applying postoperative pressure to the ear lobe. This is fabricated with cold-cure poly methyl methacrylate and orthodontic wire. This has been used in 41 ear lobe keloids in 26 patients along with postexcisional triamcinolone acetonide therapy, 5 ear lobe reconstructions and 2 cases of ear lobe clefts over a period of 8 years. Although these clips are not very aesthetic, still the acceptability is very high as the patients were well motivated. A total of 27 ear lobes in 18 patients of ear lobe keloids have undergone ear boring after 6-18 months of pressure therapy. The only complication noticed is in the form of pressure ulcer due to continuous pressure in two patients at the beginning of our experience with these clips. These healed well after temporarily stopping the use of the clip.

Ear, External↗

Thromboxane-B2, prostaglandin-E2 and hypertension in the rat 2-kidney 1-clip model: a possible mechanism of the garlic induced hypotension.

Serum collected from unilaterally clipped and unclipped rats before and after treatment with water, garlic or cilazapril and subsequent to measuring blood pressure was assayed for thromboxane-B2 and prostaglandin-E2. The unclipped rats' thromboxane-B2 and prostaglandin-E2 levels were about 23 ng/ml and 2 ng/ml, respectively, and blood pressure was 126+/-3 mmHg. These values were not affected by either water or garlic administration. The clipped rats' thromboxane-B2 and prostaglandin-E2 concentrations were close to 34 ng/ml and 4 ng/ml, respectively, and declined only in response to garlic (by 15 ng/ml and 3 ng/ml) and cilazapril (by 12 ng/ml and 1.5 ng/ml). The blood pressure of these rats was 196+/-7 mmHg and again was reduced only by garlic to 169+/-14 mmHg and cilazapril to 137+/-5 mmHg. The no-treatment and water-treatment readings were significantly higher in the clipped rats. The data suggest that prostanoid system activity in the 2-kidney 1-clip rat is enhanced and mostly toward maintaining the hypertension. Furthermore, the blood pressure lowering effects of garlic and cilazapril might have been induced partially by a greater reduction in the synthesis of vasoconstrictor prostanoids.

Animals↗

End-to-side anastomoses in small arteries with partial lateral clipping. An experimental study.

The problem of total therapeutic occlusion of the cranial arteries remains a relatively important area of concern for neurosurgery, neurology, neuroradiology and neuroanesthesia as well as for the patients. Cerebral revascularization procedures require total occlusion of the cranial arteries for a while. End-to-side anastomoses with partial lateral clipping were done in 20 cadaver arteries and then, in one rat common carotid artery. The results of the study were compared with the current cerebrovascular bypass procedures on a theoretical basis and possible clinical implications of the method were suggested. All the anastomoses were found to be open. As conclusions; 1) End-to-side anastomosis with partial lateral clipping and along with this technique the maintenance of blood flow in the target vessel during the anastomosis procedure are experimentally possible. 2) Although the available aneurysm clips may be used in the bigger vessels (> 2.5 mm), currently, the exclusive clips for the technique are not available commercially. 3) Further clinical implications of the technique may be investigated.

Anastomosis, Surgical↗

[Spiral CT angiography in demonstration of cerebral aneurysms before and after treatment with titanium clips].

PURPOSE: To determine the suitability of spiral CT angiography (SCTA) in patients with intracerebral titanium aneurysm clips. MATERIAL AND METHOD: The section parameters were optimised using a phantom. 16 patients were examined preoperatively and 18 following treatment of aneurysms with clips. RESULTS: The resolution, sensitivity and specificity for demonstrating cerebral aneurysms measuring > or = 3 mm was 91.3% and 75% respectively. In 15 patients with intracerebral clips there were no or only minor artefacts; three examinations were of no value because of major metal artefacts. Amongst six aneurysms demonstrated angiographically, two, which measured more than 3 mm, could be shown by SCTA. CONCLUSION: SCTA was able to demonstrate intracerebral aneurysms measuring > or = 3 mm. Titanium clips did not usually affect image quality significantly.

Adolescent↗

[Artefacts in the CT scan caused by surgical clips and central venous catheters and the use of new materials].

When metal clips and venous catheters have been used the computerized tomography shows artifacts, which make it computerized tomography shows artifacts, which make it difficult to have a reliable and accurate diagnosis, because the X- raying of the metal clips and the venous catheter gives radiating lines, so called streaking. We have tested new materials for the metal clips and a new polymer to replace the substance of the venous catheter with the intention that the materials should have the same density as the normal tissues. With a new metal for the clips and a new polymer for the venous catheters, mixed with metal ions, the computerized tomography does not show streaking.

Brain↗

The Filshie clip for female sterilization: a review of world experience.

Laparoscopic tubal electrocoagulation continues to be widely practiced; however, mechanical devices such as the Yoon band, the Hulka clip, and, most recently, the Filshie clip are becoming more popular because of the avoidance of accidental electrical burns, the diminished likelihood of subsequent ectopic pregnancy, and, in the case of the clips, the minimal degree of tubal destruction, thus allowing for maximum reversibility. This survey of worldwide reports from 1981 to the present reveals a high level of acceptance of the Filshie clip because of its effective design and ease of application.

Adult↗

Early experience of coronary artery bypass grafting with a new self-closing clip device.

OBJECTIVE: We are in the midst of development of several new anastomotic devices for use in coronary artery bypass grafting surgery. This study was designed to examine one of these devices (a new self-closing clip) for left internal thoracic artery-left anterior descending coronary artery anastomosis. Its feasibility and the quality of anastomosis were evaluated. METHODS: Fourteen patients who underwent first-time elective coronary artery bypass surgery were enrolled between July and December 2000. The left internal thoracic artery was anastomosed to the left anterior descending coronary artery in an interrupted manner with Coalescent U-clips (Coalescent Surgical, Inc, Sunnyvale, Calif). Immediate patency was checked with a transit-time flowmeter. Selective angiography was performed 6 months after surgical intervention. RESULTS: Five patients underwent on-pump coronary bypass grafting, 9 on the beating heart. One patient was excluded from the study intraoperatively because of a poor target site necessitating a 2-cm-long anastomosis. Left internal thoracic artery-left anterior descending artery anastomoses were created with an average of 11.8 clips in 15.9 minutes. Mean graft flow was 45.6 mL/min. Neither conversion to standard suture technique nor revision of anastomosis was necessary. There tended to be a learning curve in the anastomosis on the beating heart. Postoperative lengths of stay in the intensive care unit and the hospital were 20.7 hours and 3.9 days, respectively. Neither death nor major complication was seen, except for temporary atrial fibrillation in 2 patients. Graft patency at 6 months was 100% (FitzGibbon grade A). CONCLUSION: Left internal thoracic artery-left anterior descending artery anastomoses can be created safely and effectively with new self-closing clips on the beating, as well as the arrested, heart. Midterm patency was shown to be perfect by means of angiography.

Adult↗

Role of neuropeptide Y in the development of two-kidney, one-clip renovascular hypertension in the rat.

OBJECTIVE: Along with the renin-angiotensin system, sympathetic stimulation may contribute to renovascular hypertension. The vasoactive peptide neuropeptide Y (NPY) is co-released with and potentiates the pressor effects of norepinephrine through the Y-1 receptor. NPY, by exaggerating sympathetic activity, may contribute to renovascular hypertension, possibly by augmenting adrenergic-mediated renin release. This was studied by determining the effect of continuous Y-1 blockade on the development of two-kidney, one-clip renovascular hypertension and the effect of NPY on in vitro renin release. METHODS: Mean arterial pressure and renal blood flow responses to NPY (10 microg/kg, administered intravenously) were measured in five anesthetized Sprague-Dawley rats before and after BIBO3304TF administration to test the Y-1 antagonist BIBO3304TF. In hypertension studies, 28 rats underwent left renal artery clipping. Of these, 13 were implanted with a mini-osmotic pump for continuous BIBO3304TF infusion (0.3 microg/h, administered intravenously); the other 15 underwent sham implantation. Systolic blood pressure was then monitored for 4 weeks. Finally, in vitro renin release was measured from renal cortical slices (n = 6-12) incubated with NPY (10(-8) to 10(-6) mol/L) or NPY plus the adrenergic agonist isoproterenol (10(-4) mol/L). RESULTS: BIBO3304TF attenuated the NPY-induced increase in mean arterial pressure by 54% (P <.02) and the NPY-induced decrease in renal blood flow by 38% (P <.05). In 4-week hypertension studies, systolic blood pressure in clipped controls increased from 130 +/- 3 mm Hg to 167 +/- 6 mm Hg (P <.01), whereas BIBO3304TF-treated rats had no significant increase (125 +/- 3 mm Hg to 141 +/- 8 mm Hg). Final systolic blood pressure was 26 mm Hg lower in BIBO3304TF-treated rats than in controls (P <.01). In renal cortical slices, no NPY effect was observed in basal or isoproterenol-stimulated renin release. CONCLUSIONS: The Y-1 receptor antagonist BIBO3304TF attenuated acute pressor responses to NPY and blunted the development of two-kidney, one-clip renovascular hypertension in rats. NPY may contribute to the hypertensive response in this renovascular hypertension model. Our in vitro data do not suggest that this is due to NPY enhancement of renin release.

Animals↗

Protease specificity determination by using cellular libraries of peptide substrates (CLiPS).

We report a general combinatorial approach to identify optimal substrates of a given protease by using quantitative kinetic screening of cellular libraries of peptide substrates (CLiPS). A whole-cell protease activity assay was developed by displaying fluorescent reporter substrates on the surface of Escherichia coli as N-terminal fusions. This approach enabled generation of substrate libraries of arbitrary amino acid composition and length that are self-renewing. Substrate hydrolysis by a target protease was measured quantitatively via changes in whole-cell fluorescence by using FACS. FACS enabled efficient screening to identify optimal substrates for a given protease and characterize their cleavage kinetics. The utility of CLiPS was demonstrated by determining the substrate specificity of two unrelated proteases, caspase-3 and enteropeptidase (or enterokinase). CLiPS unambiguously identified the caspase-3 consensus cleavage sequence DXVDG. Enteropeptidase was unexpectedly promiscuous, but exhibited a preference for substrates with the motif (D/E)RM, which were cleaved substantially faster than the canonical DDDDK recognition sequence, widely used for protein purification. CLiPS provides a straightforward and versatile approach to determine protease specificity and discover optimal substrates on the basis of cleavage kinetics.

Amino Acid Sequence↗

Are clips useful in the surgical repair of colonic perforations?

PURPOSE: The objective of this study was to assess the effecacy of clip application in the surgical repair of iatrogenic colonic perforations. MATERIAL AND METHODS: Twenty adult male Wistar-Albino rats were divided into two equal groups of ten. In both of the groups an iatrogenic perforation was made in the anti-mesenteric border of the left colon. The defect was closed with interrupted 6/0 polypropylene sutures in group 1 and extraluminal application of vascular clips (VCS--vascular clip system) in group 2. All animals were killed on postoperative Day 4. Wound healing was evaluated with both in situ bursting pressure and hydroxyproline levels. RESULTS: There was no statistically significant difference between the two groups in respect of bursting pressure levels (p = 0.063) whereas hydroxyproline levels were higher in group 2 (p = 0.0021). CONCLUSIONS: Surgical repair of iatrogenic colonic perforations by extraluminal clips gives comparable results according to wound healing parameters. This approach may be a rational alternative to suture or endoscopic repair methods.

Animals↗

CLIPR-59, a new trans-Golgi/TGN cytoplasmic linker protein belonging to the CLIP-170 family.

The microtubule cytoskeleton plays a fundamental role in cell organization and membrane traffic in higher eukaryotes. It is well established that molecular motors are involved in membrane-microtubule interactions, but it has also been proposed that nonmotor microtubule-binding (MTB) proteins known as CLIPs (cytoplasmic linker proteins) have basic roles in these processes. We report here the characterization of CLIPR-59, a CLIP-170-related protein localized to the trans-most part of the Golgi apparatus. CLIPR-59 contains an acidic region followed by three ankyrin-like repeats and two CLIP-170-related MTB motifs. We show that the 60-amino acid-long carboxy-terminal domain of CLIPR-59 is necessary and sufficient to achieve Golgi targeting, which represents the first identification of a membrane targeting domain in a CLIP-170-related protein. The MTB domain of CLIPR-59 is functional because it localizes to microtubules when expressed as a fragment in HeLa cells. However, our results suggest that this domain is normally inhibited by the presence of adjacent domains, because neither full-length CLIPR-59 nor a CLIPR-59 mutant missing its membrane-targeting region localize to microtubules. Consistent with this observation, overexpression of CLIPR-59 does not affect the microtubule network. However, CLIPR-59 overexpression strongly perturbs early/recycling endosome-TGN dynamics, implicating CLIPR-59 in the regulation of this pathway.

Amino Acid Sequence↗

pH-dependent peptide binding properties of the type I diabetes-associated I-Ag7 molecule: rapid release of CLIP at an endosomal pH.

MHC class II molecules and invariant chain assemble at a neutral pH in the endoplasmic reticulum and are transported to a low pH compartment where the invariant chain is trimmed to the class II-associated invariant chain peptide (CLIP). For many major histocompatibility complex class II molecules, DM is required for rapid removal of CLIP, which allows binding of antigenic peptides. Since I-Ag7 confers susceptibility to type I diabetes in NOD mice, the biochemical requirements for peptide loading were examined using soluble I-Ag7 expressed in insect cells. I-Ag7 formed long-lived complexes with naturally processed peptides from transferrin and albumin, whereas several peptides that represent T cell epitopes of islet autoantigens were poor binders. I-Ag7-peptide complexes were not sodium dodecyl sulfate (SDS) resistant, indicating that SDS sensitivity may be an intrinsic property of I-Ag7. Complexes of I-Ag7 and CLIP formed at a neutral pH, but rapidly dissociated at pH 5. This rapid dissociation was due to a poor fit of M98 of CLIP in the P9 pocket of I-Ag7, since substitution of M98 by a negatively charged residue greatly enhanced the stability of the complex. These biochemical properties of I-Ag7 result in the rapid generation of empty molecules at an endosomal pH and have a global effect on peptide binding by I-Ag7.

Amino Acid Sequence↗