Qualitative study of fecal alpha 1-proteinase inhibitor in normal subjects and patients with Crohn's disease

We applied sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting to analyze fecal alpha 1-proteinase inhibitor (alpha 1 PI) from healthy subjects and patients with Crohn's disease. A component with Mr 38,000 was characterized in normal fecal extracts as well as in six pathological samples. In these cases, the Crohn's disease activity index (CDAI), a clinical index of severity of the disease, was 170 (SEM 47). In contrast, alpha 1 PI of Mr 51,000 was detected in fecal extracts from eight patients with active Crohn's disease (CDAI = 287, SEM 39). We conclude that fecal alpha 1 PI can be considered a marker of intestinal disease activity.     

Association between mutations in the CARD15 (NOD2) gene and Crohn's disease in Israeli Jewish patients

Ulcerative colitis (UC) and Crohn's disease (CD) are heterogeneous disorders characterized by chronic intestinal inflammation. Genetic predisposition is a major risk factor in both diseases. The CARD15 (NOD2) gene has been implied as a candidate gene in the pathogenesis CD. Our aim was to delineate the frequency of three missense and one frameshift variant of CARD15 in Israeli Jewish CD and UC patients. DNA was extracted from blood samples from 238 unrelated inflammatory bowel disease (IBD) patients, 68 with UC and 170 with CD. The DNA was genotyped for two missense mutations, R675W and G881R, and one frameshift mutation, 980FS981X. Mutations in CARD15 were observed with significantly greater frequency in CD patients (46/170, 27%) than in UC patients (7/68, 10%) (P = 0.005). Homozygous and compound heterozygous carriers were restricted to seven (4%) patients with CD as compared to none of the UC patients (P = 0.01). Similar rates in Ashkenazi and non-Ashkenazi Jewish patients were observed. Age-of-onset of disease was lower in Ashkenazi mutation carriers as compared to non-carriers of Ashkenazi origin (18.7 +/- 8.6 years vs. 25.8 +/- 13.4 years, respectively, P = 0.03). No other phenotypic characteristics could distinguish mutation carriers from non-carriers. We conclude that germline mutations in the CARD15 gene are more frequently found in CD than UC patients and appear to predict an earlier age-of-onset in Ashkenazi Jewish patients. No association could be demonstrated between CARD15 mutations and specific disease course or behavior.     

Eosinophilic gastroenteritis: ultrastructural evidence for a selective release of eosinophil major basic protein.

Ultrastructural study of mucosal eosinophils in a case of eosinophilic gastroenteritis involving stomach, duodenum and ileum showed an altered structure in ulcerated duodenal areas. The electron core density of eosinophil granules was inverted or disappeared and tubulovesicular structures occurred. Using immunogold staining with specific antibodies, major basic protein was detected diffusely in the matrix of eosinophil granules and out of the granules in tight association with extragranular membrane formations. In contrast, eosinophil cationic protein and eosinophil peroxidase were normally distributed in the granule matrix. When compared with the eosinophils in macroscopically normal duodenal mucosa in the same patient, these changes support a role for major basic protein in tissue damage in eosinophilic gastroenteritis. The diffusion of one granule protein from the granules to the exterior of the cells favours the view of a selective release of eosinophil mediators.     

Detection of the apoptosis-suppressing oncoprotein bc1-2 in hormone-refractory human prostate cancers.

The oncoprotein encoded by bc1-2 is unique because of its intracellular location (a mitochondrial membrane protein) and apparent mode of action (suppression of apoptosis). To date, this oncogene has been associated only with the development of certain forms of human B-cell lymphoma. In this report, we describe our experience with a monoclonal antibody made against a synthetic peptide for bc1-2 that can recognize the bc1-2 protein and identify cells in human prostate glands expressing this proto-oncogene with in situ immunohistochemical procedures. These procedures were utilized to survey a series of 62 human tissues to evaluate whether bc1-2 might have a role in the developing prostate gland or in prostate oncogenesis. While all primordial epithelial cells in a fetal prostate gland immunostain for bc1-2, normal and hypertrophic prostate glands of the adult show bc1-2 expression restricted to the basal cells. All epithelial cells in areas of prostatic intraepithelial neoplasia were stained by this antibody, as were most (62%) localized invasive prostatic carcinomas. In contrast, all primary prostatic carcinomas and metastases obtained from metastatic prostate cancer patients after hormone treatment (hormone-refractory tumors) stained positive for bc1-2. This study demonstrates that the oncoprotein encoded by bc1-2 can be detected at sequential stages in the natural history of human prostate cancer. Since the bc1-2 oncoprotein is known to suppress the cellular response to apoptotic stimuli, it will be important to determine whether bc1-2 expression is a factor in the development of prostate cancers and in the survival of hormone-refractory prostate cancer cells.     

Are HLA-DR or TAP Genes Genetic Markers of Severity in Ulcerative Colitis?

The pathogeny of ulcerative colitis (UC) is not yet elucidated, but some arguments suggest the implication of genetic factors. Among the candidate genes, those encoding for HLA class II genotypes have been extensively studied in UC; however, discordant data may be imputable to heterogeneity, characterized by immunological markers such as atypical ANCA (p-ANCA), or to inclusion of more or less intractable UC. The aim of our study is to evaluate the interest of HLA class II and TAP genetic markers to identify different clinical forms of UC, according to p-ANCA status. Unrelated patients with a history of UC (n=91) and healthy control subjects with no personal or family history of inflammatory bowel diseases (IBD) (n=200) were included. HLA-DRB103 was less frequent in UC patients than in healthy controls (8% vs 28%,PC<0.03). No association was found with any TAP genotypes. Moreover, there was no association with the HLA-DR2 specificity, either in the entire group of UC patients (38% vs 28%) or in the p-ANCA-positive subgroup of patients (30%). The most consistent finding in the present study is that some genetic markers may characterize intractability in UC patients. HLA-DR2 was associated with poor prognosis, regardless of p-ANCA status. In HLA-DR2 and non-HLA-DR2 groups, colectomy was done in 55% and 27% of patients, respectively (PC<0.05). Furthermore, in non-HLA-DR2 patients, p-ANCA could be of interest to characterize those with more severe prognosis. Our results confirm the interest of genetic studies to define UC genetic susceptibility, taking into account intractability of the disease. They do not support the hypothesis that p-ANCA is a subclinical marker of genetic susceptibility to UC.     

Arterial and lymphatic supply of the knee integuments

Summary The surgical approach to the anterior knee region carries a risk for postoperative integument infection and skin necrosis. A hypothesis is that surgical approach may damage integumental blood supply and additionally the lymphatic drainage from the foot and leg. The goal of this study was to describe the arteries and lymphatics directly affected by the antero-medial approach. Injection of the femoral a. was used to identify the femoral and popliteal aa. and their branches distributed to the integument of the anterior, medial and lateral aspects of the knee. Lymphatic injection into the plantar aspect of the first toe was also performed to identify the subdermal lymphatics traversing the area. Our results showed that most of the blood supply arises from the medial aspect of the knee integuments. However, subcutaneous arterial anastomoses provide a significant blood-supply when there is interruption of the medial vessels as seen in the medial surgical approach to the knee. Most of the lymphatic drainage originating from the foot crosses the knee region on the medial side, opposite or below the tibial tuberosity. Because the subdermal arterial network is well-developed, the medial approach for knee surgery does not endanger the anterior knee integuments as long as the lateral vascular supply is preserved. However, this approach may interrupt the lymphatic circulation, particularly in the case of an extended incision, which could explain postoperative edema and an increased rate of wound infection.

---

Vascularisation artérielle et lymphatique des téguments du genou

Résumé L'abord chirurgical du genou présente un risque d'infection post-opératoire et de nécrose cutanée. Une hypothèse serait que la voie d'abord peut détruire la vascularisation artérielle et lymphatique qui draine la jambe et le pied. Le but de notre étude était de décrire la circulation artérielle et lymphatique impliquée dans la voie d'abord antéromédiale du genou. Des injections ont été utilisées pour identifier les aa. fémorale et poplitée et leurs branches qui se distribuent aux téguments de la région médiale et latérale du genou. Des injections lymphatiques ont été également utilisées pour localiser le réseau subdermique lymphatique de la région du genou. Nos résultats montrent que la majorité de l'apport artériel des téguments du genou provient de la partie médiale. Des anastomoses sub-cutanées entre les systèmes artériels latéral et médial permettent un apport artériel suffisant pour les téguments en cas d'interruption de l'apport médial. La plupart des vaisseaux lymphatiques provenant du pied et de la cheville croisent la région dorsale du pied de dehors en dedans à une distance variable de la tubérosité tibiale. Du fait du bon développement du réseau artériel sub-cutané, la voie d'abord médiale ne présente pas de risque de nécrose des téguments de la face antérieure du genou tant que l'apport latéral est préservé. Toutefois, elle peut être responsable de l'interruption des conduits lymphatiques, en particulier lorsque l'incision s'étend vers le bas, exposant aux lésions des collecteurs lymphatiques, ce qui expliquerait un dème post-opératoire et un taux accru d'infection.

     

Results of pyeloureterostomy after ureterovesical anastomosis complications in renal transplantation

OBJECTIVES: The most frequent urologic complications after renal transplantation involve the ureterovesical anastomosis (ie, leakage, stenosis, and reflux), with a frequency of 1% to 30% in different series. We present the results of pyeloureterostomy using the recipient's ureter. METHODS: From 1988 to 1996, 570 cadaveric renal grafts were performed at our institution. A Lich Gregoir ureterovesical anastomosis was used in every case. Complications involving the anastomosis occurred in 19 cases (3.3%), with 10 stenoses (1.7%), 6 cases of leakage (1.1%), and 3 of reflux (0.5%). The mean donor age was 36.2 years, and the mean duration of cold ischemia was 29.4 hours. The mean recipient age was 41.3 years. Corrective surgery was performed 0.09 years (range 0.01 to 0.22) after transplantation for leakage, 1.13 years (range 0.14 to 5.11) for stenosis, and 5.55 years (range 0.51 to 9.71) for reflux. The recipient's ureter was stented with a ureteral catheter before median laparotomy, except in 3 cases of early leakage (less than 3 days). The recipient's ureter was cut, without the need for ipsilateral nephrectomy, and sutured to the graft pelvis. A nephroureterostomia stent (Gil Vernet stent) (12 cases) or a double J ureteral stent (7 cases) was used for urinary drainage. RESULTS: One graft was lost on day 1 through renal vein thrombosis. Percutaneous nephrostomy was performed on day 2 to clear an obstruction of the double J ureteral stent in one case, and a double J ureteral stent was inserted on day 2 because the nephrouretrostomia stent was incorrectly positioned in another case. Pyelographic controls on day 15 were normal in every case. The mean follow-up was 2.25 years (range 0.24 to 6.1) (2.9 years for leakage, 2.08 years for stenosis, and 1.44 years for reflux). One patient died with a functional graft 3 years after surgery. One graft was lost 4 years after surgery through chronic rejection. There were no complications affecting the ipsilateral kidney. No further ureteral complications occurred after surgery. The mean creatinine level 3 years after surgery was 1.59 mg/dL. CONCLUSIONS: Pyeloureterostomy is a safe and permanent treatment for complications of ureterovesical anastomosis and gives excellent results. The technique requires stenting of the recipient's ureter and graft drainage with a nephroureterostomia stent or a double J ureteral stent.     

Digestive manifestations of food hypersensitivity in adults]

Food allergy, synonymous with food hypersensitivity (FHS), is defined as an immunologically-mediated adverse reaction to food. Initiation of FHS could result from a break in the immune mucosal barrier with abrogation of oral tolerance. Food hypersensitivity is mostly due to immediate-type reaction involving IgE-dependent mastocytes activation. Changes in intestinal function and structure have been mainly studies in an animal model of rat sensitized to egg albumin. Intraluminal antigen challenge resulted in abnormalities of gut absorption, secretion and motility in sensitized rats. In man, experimental data are scarce. Gastrointestinal manifestations of immediate FHS are varying and unspecific. A role for FHS in irritable bowel syndrome is debated. Participation of delayed-type FHS to digestive diseases is still questionable, but eosinophilic gastroenteritis might be an example. In clinical practice, diagnosis of FHS demands rigorous criteria. Double blind placebo-controlled food challenge has eventually proved to be the "gold standard" test for FHS diagnosis.     

Précipitation et prévention de l'abstinence chez le rat et la souris en état de dépendance aiguë. Comparaison de la naloxone,de la naltrexone et de la diprenorphine

Summary Abstinence signs were precipitated in rats by naloxone (1 mg·kg^-1 s.c.) injected at various times (from 1.5 to 16 h) after a single dose of morphine hydrochloride (15 or 50 mg·kg^-1 s.c.) administered incaqueous solution. Increasing the dose of morphine increased the latency of the phenomena and the duration of the underlying state shifts of signs as described by Bläsig et al. (1974) in chronically morphinized rats also occurred when increasing the dose of morphine and the time interval between the injections of morphine and of naloxone. Naltrexone and diprenorphine were also effective. These three antagonists, given before morphine, were able to prevent precipitated abstinence: however, naloxone was almost ineffective when the higher dose of morphine was used and when the time interval was long. In these latter conditions, naltrexone was definitely more effective and longer acting and diprenorphine still more so. The same characteristics were found for the protective action of the three antagonists in acutely morphinized mice and the same order for their potencies in precipitating abstinence in acutely morphinized mice. Like naloxone, naltrexone and diprenorphine facilitated a nociceptive reaction in normal mice.The abstinence signs precipitated in acutely morphinized rats or mice are probably not unmasked excitatory effects of morphine as such effects should have been increased rather than inhibited by previous administration of specific antagonists; they might correspond to potentiated effects of the antagonists themselves. The prevention by specific antagonists of the abstinence syndrome is most simply interpreted by antagonism (direct or indirect) of dependence induction, but other interpretations are not excluded.