Prognostic value of increased intraepithelial lymphocytes and lymphocytic clonality in dogs with chronic enteropathy or small-cell lymphoma.

The clinical significance of severe infiltration of small intraepithelial lymphocytes (IEL) and the results of polymerase chain reaction for antigen receptor rearrangement (PARR) in dogs with chronic enteropathy (CE) and small-cell lymphoma (SCL) are controversial. This cohort study aimed to evaluate the prognostic significance of the IEL and PARR results in dogs with CE or SCL. Although definitive diagnostic histopathological criteria for SCL in dogs have yet to be established, dogs with the histopathological findings of severe IEL infiltration were diagnosed with SCL in this study. One hundred and nineteen dogs were recruited, with 23 dogs classified as having SCL and 96 dogs as having CE. The positive rate of PARR was 59.6 % (71/119) in the duodenum and 57.7 % (64/111) in the ileum. Subsequently, three dogs with SCL and four dogs with CE developed large-cell lymphoma (LCL). The median overall survival (OS) of dogs with SCL was 700 days (range, 6-1410 days), and that of dogs with CE was not reached. In the log-rank test, shorter OS was observed in cases with histopathological SCL (P = 0.035), clonal TCRγ rearrangement in the duodenum (P = 0.012), and clonal IgH rearrangement in the ileum (P < 0.0001). The Cox proportional hazards model adjusted for sex and age showed that histopathological SCL (hazard ratio [HR] 1.74; 95 % confidence interval [CI], 0.83-3.65), duodenal clonal TCRγ rearrangement (HR, 1.80; 95 % CI, 0.86-3.75), and ileal clonal IgH rearrangement (HR, 2.28; 95 % CI, 0.92-5.70) could shorten overall survival, although their 95 % CIs included 1.0. These results indicate that severe IEL infiltration could be a useful histopathological feature for diagnosing SCL, and clonality-positive results could be a negative prognostic factor in dogs with CE. Furthermore, the development of LCL should be carefully monitored in dogs with CE and SCL..

Endoscopic Cytology for the Diagnosis of Chronic Enteritis and Intestinal Lymphoma in Dogs.

Although cytology is a rapid diagnostic procedure in dogs, the cytologic criteria of endoscopic biopsies for chronic enteritis and intestinal lymphoma are not well defined. An immediate diagnosis using cytology would benefit patients by enabling prompt initiation of therapy. The objective of this study was to investigate the correlation between the results of endoscopic cytology and histopathology. In this study, 167 dogs with clinical signs of chronic gastrointestinal disease were included. On the basis of histopathology, the following diagnoses were determined: lymphocytic-plasmacytic enteritis in 93 dogs; eosinophilic enteritis in 5 dogs; small cell intestinal lymphoma in 45 dogs; and large cell intestinal lymphoma in 24 dogs. Two clinical pathologists retrospectively evaluated the endoscopic cytology of squash-smear preparations. The cytologic diagnoses of inflammation, small cell lymphoma, and large cell lymphoma were based on the severity of lymphocyte infiltration, the size of infiltrated lymphocytes, and eosinophil/mast cell infiltration. The clinical severity score was significantly increased along with the degree of lymphocyte infiltration evaluated by cytology. The cytologic diagnosis was in complete agreement with the histopathologic diagnosis in 136 of 167 (81.4%) cases. For the differentiation between enteritis and lymphoma, endoscopic cytology had a sensitivity of 98.6%, a specificity of 73.5%, a positive predictive value of 72.3%, and a negative predictive value of 98.6%. The log-rank test and Cox regression analysis showed that the results of cytology predicted the prognosis. These results suggest that endoscopic cytology is a useful technique to aid diagnosis of intestinal inflammation and lymphoma in dogs.

Proliferation, differentiation and amyloid-ß production in neural progenitor cells isolated from TgCRND8 mice.

The amyloid precursor protein (APP) and amyloid-ß (Aß) peptide play central roles in the pathology and etiology of Alzheimer's disease. Amyloid-induced impairments in neurogenesis have been investigated in several transgenic mouse models but the mechanism of action remains to be conclusively demonstrated. The changes in neurogenesis during this transition of increasing Aß levels and plaque formation were investigated in the present study. We found that the proliferation of newborn cell in the dentate gyrus was enhanced prior to elevations in soluble Aß production as well as amyloid deposition in 5-week-old TgCRND8 mice, which are well-established Alzheimer's disease models, compared to non-transgenic (Non-Tg) mice. The number of BrdU-positive cells remained higher in TgCRND8 vs Non-Tg mice for a period of 8weeks. The numbers of BrdU/NeuN-positive cells were not significantly different in TgCRND8 compared to Non-Tg mice. A significant decrease in BrdU/GFAP but not in BrdU/S100ß was found in Tg vs Non-Tg at 6-weeks of age. In addition, a unique observation was made using isolated neuroprogenitor cells from TgCRND8 mice which were found to be less viable in culture and produced substantial amounts of secreted Aß peptides. This suggests that the proliferation of neural progenitors in vivo may be modulated by high levels of APP expression and the resulting Aß generated directly by the progenitor cells. These findings indicate that cell proliferation is increased prior to Aß deposition and that cell viability is decreased in TgCRND8 mice over time.

Activation of OASIS family, ER stress transducers, is dependent on its stabilization.

Endoplasmic reticulum (ER) stress transducers transduce signals from the ER to the cytoplasm and nucleus when unfolded proteins accumulate in the ER. BBF2 human homolog on chromosome 7 (BBF2H7) and old astrocyte specifically induced substance (OASIS), ER-resident transmembrane proteins, have recently been identified as novel ER stress transducers that have roles in chondrogenesis and osteogenesis, respectively. However, the molecular mechanisms that regulate the activation of BBF2H7 and OASIS under ER stress conditions remain unresolved. Here, we showed that BBF2H7 and OASIS are notably unstable proteins that are easily degraded via the ubiquitin-proteasome pathway under normal conditions. ER stress conditions enhanced the stability of BBF2H7 and OASIS, and promoted transcription of their target genes. HMG-CoA reductase degradation 1 (HRD1), an ER-resident E3 ubiquitin ligase, ubiquitinated BBF2H7 and OASIS under normal conditions, whereas ER stress conditions dissociated the interaction between HRD1 and BBF2H7 or OASIS. The stabilization of OASIS in Hrd1(-/-) cells enhanced the expression of collagen fibers during osteoblast differentiation, whereas a knockdown of OASIS in Hrd1(-/-) cells suppressed the production of collagen fibers. These findings suggest that ER stress stabilizes OASIS family members and this is a novel molecular mechanism for the activation of ER stress transducers.

A molecular chaperone inducer protects neurons from ER stress.

The endoplasmic reticulum (ER) stress response is a defense system for dealing with the accumulation of unfolded proteins in the ER lumen. Recent reports have shown that ER stress is involved in the pathology of some neurodegenerative diseases and cerebral ischemia. In a screen for compounds that induce the ER-mediated chaperone BiP (immunoglobulin heavy-chain binding protein)/GRP78 (78 kDa glucose-regulated protein), we identified BiP inducer X (BIX). BIX preferentially induced BiP with slight inductions of GRP94 (94 kDa glucose-regulated protein), calreticulin, and C/EBP homologous protein. The induction of BiP mRNA by BIX was mediated by activation of ER stress response elements upstream of the BiP gene, through the ATF6 (activating transcription factor 6) pathway. Pretreatment of neuroblastoma cells with BIX reduced cell death induced by ER stress. Intracerebroventricular pretreatment with BIX reduced the area of infarction due to focal cerebral ischemia in mice. In the penumbra of BIX-treated mice, ER stress-induced apoptosis was suppressed, leading to a reduction in the number of apoptotic cells. Considering these results together, it appears that BIX induces BiP to prevent neuronal death by ER stress, suggesting that it may be a potential therapeutic agent for cerebral diseases caused by ER stress.

[Surgical techniques and results of right ventricular [correction of venericular] outflow tract reconstruction with pedicled autologous tissue].

From 1987 to 2000, 44 patients underwent anatomic repair of anomalies with pulmonary outflow obstruction by using pedicled autologous tissue in reconstructing the pulmonary outflow tract. Twenty-four patients had tetralogy of Fallot with pulmonary atresia (TOF/PA), from 1.7 years to 12.9 years of age (mean 9.0 years). In the group of TOF/PA, slit-widening method was performed in 22, autologous pericardial flap was used in 1, and atrial appendage was used in 4 patients. Eight patients had transposition of the great arteries with ventricular septal defect and pulmonary stenosis (TGA type III), from 9 months to 9.2 years of age (mean 4.5 years). All of the patient with TGA type III under went direct PA-RV anastomosis (natural route non-conduit repair). Three patients had truncus arteriosus, from 1 month to 7.8 years of age (mean 4.5 years). In the group of the truncus, atrial appendage was used in 1 and direct PA-RV anstomosis was made in 2 patients. Six patients had Rastelli's conduit obstruction, from 6.6 years to 20.3 years of age (mean 13.9 years). The conduit was removed and outflow tract was reconstructed with direct PA-RV anastomosis in 5 and using atrial appendage in 1 patients. There was one hospital death (2%) and no late deaths. In 31 patients whose follow-up time exceed 5 years, 7 patients (23%) were found to have pressure gradient more than 40 mmHg at the pulmonary outflow tract. None of the patients with TGA who underwent natural route non-conduit repair developed late stenosis at the pulmonary out flow tract.

[Hemodynamic effects of amrinone or milrinone in infant after right heart bypass operation].

After right heart bypass operation, dopamine was continuously infused at 5 micrograms/kg/min. The patients had a stable hemodynamic status but relatively low SvO2, low urine output and peripheral insufficiency. Amrinone was continuously infused at 10 micrograms/kg/min in 5 patients (Group A), or milrinone was continuously infused at 0.5 microgram/kg/min in 5 patients (Group M). After infusion of amrinone or milrinone, arterial blood pressure relatively decreased and heart rate relatively increased. But heart work index did not increased. SvO2 relatively increased. Mean pulmonary artery pressure relatively decreased. The administration of amrinone (Group A) produced relatively thrombocytopenia compared with milrinone (Group M). Two patients had supraventricular arrhythmia in Group A. A patient had ventricular arrhythmia and another patient supraventicular arrhythmia. These patients treated using temporary antiarrhythmia drug or temporary pacing. When the patients discharged, they recovered the normal sinus rhythm. In conclusion, after right heart bypass operation in infants, infusion of amrinone or milrinone appears to be effective and safe combining with dopamine.

Anastomosis of the left juxtaposed atrial appendages in a patient with tricuspid atresia.

A 9-month-old boy with left juxtaposition of the atrial appendages, tricuspid atresia, pulmonary atresia, and ventriculoarterial discordance underwent anastomosis between the atrial appendages after failure of balloon/blade atrial septostomy because of restrictive atrial septal defect. For surgical creation of atrial communication in patients with juxtaposed atrial appendages, anastomosis between the atrial appendages seemed to be safer, more effective, and less invasive than septectomy by Blalock-Hanlon technique or inflow occlusion technique.

[A case report: right ventricular outflow tract (RVOT) reconstruction using homograft for RVOT restenosis after Jatene's procedure of transposition of the great arteries (TGA)].

A 7-year-old boy was admitted for RVOT restenosis. The patient was born following an unremarkable pregnancy and delivery. He was carried out Jatene's procedure for TGA at 23rd day after birth. At 19 month of age, he was removed pulmonary stenosis using patch angioplasty with mono cusp. When he was 6 year of age, the onset of edem, ascites and hepatomegaly were pointed out. At 7 year of age, he was admitted for edema, hypoproteinemia and protein loosing enteropatchy. After he was treated medical therapy, cardiac catheterization showed markedly right atrial and right ventricular hypertension and ventricular septal defect (VSD). Cine angiography suggested the right ventricular outflow tract stenosis, pulmonary valve and tricuspid valve insufficiency. On August 1, 1996, the patient underwent surgical repair: direct closure of VSD, aortic homograft (20mm) replacement for RVOT stenosis and tricuspid valve annuloplasty by DeVega's procedure. Postoperative course was uneventful. At 23rd postoperative day, cardiac catheterization and cine angiography showed no problems. He was not treated with anticoagulation.

Right ventricular obstruction by tricuspid pouch in simple ventricular septal defect.

Tricuspid pouch sometimes causes subpulmonary obstruction in association with transposition of the great arteries, but rarely does so without transposition. A 5-year-old girl with isolated ventricular septal defect exhibited dynamic right ventricular outflow tract obstruction caused by pouches in the anterior and septal tricuspid leaflets. Echocardiography clearly demonstrated features of the tricuspid pouches. Ventricular septal defect closure resulted in disappearance of the tricuspid pouches and resolution of subpulmonary obstruction.

Repair of septal and posterior tricuspid leaflets in Ebstein's anomaly.

BACKGROUND AND AIM: In Ebstein's anomaly, the septal and posterior tricuspid leaflets are plastered to the endocardium. We postulated that tricuspid valve function could be corrected by restoring mobility of these leaflets. (Feasibility of such repair was explored by anatomical and clinical studies.) METHODS: Ten heart specimens with Ebstein's anomaly were examined to investigate the size of the tricuspid leaflets. We operated on four patients with Ebstein's anomaly: the plastered septal and posterior leaflets were mobilized from the endocardium, the atrialized right ventricle was longitudinally plicated, and the basal attachment of the mobilized leaflets was sutured (reattached) to the valve annulus. RESULTS: In heart specimens, approximately 40% of the total surface of the tricuspid leaflets was comprised of the septal and posterior leaflets. Clinically, all patients operated on returned to normal functional status after surgery. The mean cardiothoracic ratio on chest X-rays decreased from 0.70 to 0.55 (after surgery). Echocardiographic tricuspid regurgitation, graded from 0 to 4, decreased from 3.5 to 1.0, and tricuspid annular diameter ratio to the normal value reduced from 1.88 to 0.66. Angiographic right ventricular ejection fraction increased from 0.36 to 0.50, and end-diastolic volume ratio to the normal value decreased from 3.65 to 1.19. CONCLUSIONS: Repair of the septal and posterior tricuspid leaflets was found to be feasible and effective as tricuspid valvuloplasty for Ebstein's anomaly.

Simultaneous detection of Streptococcus pneumoniae and Haemophilus influenzae by nested PCR amplification from cerebrospinal fluid samples.

Haemophilus influenzae and Streptococcus pneumoniae are often the cause of serious diseases such as meningitis. We designed a nested PCR assay to identify these pathogens from cerebrospinal fluid samples. The first-step PCR was able to detect eubacterial rRNA genes with a unified set of universal primers. In the second-step PCR, the identification primers, HI I and II and SP I and II, could detect H. influenzae and S. pneumoniae respectively through amplification of the rRNA spacer between the 16S and 23S rRNA genes. We suggest that the two-step PCR assay can be used as a novel method for the immediate and retrospective diagnosis of bacterial meningitis caused by H. influenzae and S. pneumoniae.

Lay-open pulmonary arterioplasty for postoperative hilar pulmonary artery stenosis.

OBJECTIVE: Lay-open pulmonary arterioplasty, a novel surgical technique to enlarge postoperative stenosis at the hilar pulmonary artery, was evaluated. METHODS: Lay-open arterioplasty, in which the enlarged hilar stenotic pulmonary artery is partially made up of previous surgical scar tissue instead of being covered by a patch, was performed on 10 patients whose ages ranged from 2.2 to 15.7 years. Surgical results were assessed by angiography. RESULTS: All patients tolerated the procedure without bleeding or embolic complications associated with pulmonary arterioplasty. Nine patients underwent concomitant procedures including total repair (n = 5), central interposing shunt (n = 3), and right ventricular outflow tract reconstruction (n = 1). No deaths or life-threatening events occurred during the total follow-up period of 18 patient-years. The stenotic segment was significantly enlarged from the preoperative diameter of 0.9 +/- 1.1 mm (mean +/- standard deviation) to the postoperative diameter of 8.0 +/- 1.3 mm, values which correspond to 7.0% +/- 8.8% and 68.4% +/- 11.5% of the normative values, respectively. A follow-up angiogram (n = 5) revealed an increase in the pulmonary artery diameter balanced with somatic growth (initial value, 65.2% +/- 9.0% of normal; second value, 69.1% +/- 7.7% of normal). No aneurysms or clinically significant restenoses were seen on the angiograms. CONCLUSIONS: Our initial midterm results with this method were promising. The pulmonary arteries subjected to this procedure grew in proportion to somatic growth.

[Bacterial culture of the anterior chest skin for prophylaxis in cardiovascular operations: prospective study in two civilian hospitals].

Mediastinitis after median stermotomy is a significant complication for a cardiovascular surgery. Prospective study of bacterial cultures of the anterior chest skin in cardiac and aortic surgery was performed with the comparison of two civilian hospitals. There were many ABPC-resistant bacteria including 26 to 67 percent of Staphylococcus epidermidis (SE). ABPC could not be chosen for the first drug of prophylaxis. Although there was a few CEZ-resistant SE, the efficacy of first-generation cephalosporins was acceptable in this series. If methicillin-resistant Staphylococcus aureus (MRSA) is detected in patient's preoperative bacterial cultures, vancomycin should be selected for prophylaxis. In patients with poor risk who required emergent cardiac surgery, combinational use of an first-generation cephalosporin and an amikacin or a minocycline should be recommended in our data.

[Management of postoperative pulmonary hypertensive crisis in children--indication and usefulness of inhaled nitric oxide therapy as a pulmonary vasodilator].

Among 176 pediatric patients who underwent open heart surgery from 1990 to 1996, 7 developed severe pulmonary hypertensive crisis (PHC) postoperatively. All patients were treated with conventional medical therapy consisting of hyperventilation and deep sedation. Prostaglandin E1 and/or amrinone were administered initially as a pulmonary vasodilator, and in 3 of the 7 patients this resulted in immediate improvement in hemodynamics. These patients subsequently weaned off ventilatory support and they were discharged from the hospital. In 4 other patients, pulmonary vasodilator therapy with prostaglandin E1 and amrinone had failed and they were treated with inhaled nitric oxide (NO). In all patients, inhalation of NO resulted in immediate decrease in pulmonary pressure without significant change in systemic arterial pressure. Two patients successfully weaned from the ventilator, whereas in 2 patients decrease in pulmonary pressure was transient. They developed airway bleeding and died as a result of respiratory insufficiency. Although inhaled NO effectively reduced pulmonary pressure in patients with PHC, this effect was not maintained over 12 hours in patients associated with bronchopulmonary dysplasia. In patients at risk for severe PHC, rapid treatment with NO inhalation after initial event as well as the prevention of lung injury caused by mechanical ventilation were found to be important in the postoperative management.

[Inhaled nitric oxide for postoperative pulmonary hypertensive crisis in a patient with complete atrioventricular canal associated with Down's syndrome: a case report].

Two-year-old boy with Down's syndrome was treated with inhaled nitric oxide for pulmonary hypertensive crisis after correction of complete atrioventricular canal and persistent ductus arteriosus. The acute rise in pulmonary pressure and hemodynamic instability occurred on 3rd, 4th and 6th post operative days and pulmonary vasodilation with prostaglandin E1 and amrinone had failed. Twenty ppm of inhaled nitric oxide effectively reduced the pulmonary pressure and the patient was subsequently weaned from ventilation. Inhaled nitric oxide exerted a pulmonary vasodilation without decreasing systemic pressure in this patient with postoperative pulmonary hypertensive crisis.

Rapid identification of Streptococcus pneumoniae by PCR amplification of ribosomal DNA spacer region.

Streptococcus pneumoniae is one of the important human pathogens in clinical microbiology. A polymerase chain reaction assay was designed to detect and identify S. pneumoniae through amplification of the ribosomal DNA spacer regions between the pneumococcal 16S-23S ribosomal RNA genes. Thirty-two Streptococcus and non-Streptococcus strains were tested to verify the specificity of the assay, and only S. pneumoniae strains gave a positive reaction. This method is a powerful technique for the rapid identification of S. pneumoniae.