Modulation of the Wnt pathway through inhibition of CLK2 and DYRK1A by lorecivivint as a novel, potentially disease-modifying approach for knee osteoarthritis treatment.

OBJECTIVES: Wnt pathway upregulation contributes to knee osteoarthritis (OA) through osteoblast differentiation, increased catabolic enzymes, and inflammation. The small-molecule Wnt pathway inhibitor, lorecivivint (SM04690), which previously demonstrated chondrogenesis and cartilage protection in an animal OA model, was evaluated to elucidate its mechanism of action. DESIGN: Biochemical assays measured kinase activity. Western blots measured protein phosphorylation in human mesenchymal stem cells (hMSCs), chondrocytes, and synovial fibroblasts. siRNA knockdown effects in hMSCs and BEAS-2B cells on Wnt pathway, chondrogenic genes, and LPS-induced inflammatory cytokines was measured by qPCR. In vivo anti-inflammation, pain, and function were evaluated following single intra-articular (IA) lorecivivint or vehicle injection in the monosodium iodoacetate (MIA)-induced rat OA model. RESULTS: Lorecivivint inhibited intranuclear kinases CDC-like kinase 2 (CLK2) and dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A). Lorecivivint inhibited CLK2-mediated phosphorylation of serine/arginine-rich (SR) splicing factors and DYRK1A-mediated phosphorylation of SIRT1 and FOXO1. siRNA knockdowns identified a role for CLK2 and DYRK1A in Wnt pathway modulation without affecting ß-catenin with CLK2 inhibition inducing early chondrogenesis and DYRK1A inhibition enhancing mature chondrocyte function. NF-κB and STAT3 inhibition by lorecivivint reduced inflammation. DYRK1A knockdown was sufficient for anti-inflammatory effects, while combined DYRK1A/CLK2 knockdown enhanced this effect. In the MIA model, lorecivivint inhibited production of inflammatory cytokines and cartilage degradative enzymes, resulting in increased joint cartilage, decreased pain, and improved weight-bearing function. CONCLUSIONS: Lorecivivint inhibition of CLK2 and DYRK1A suggested a novel mechanism for Wnt pathway inhibition, enhancing chondrogenesis, chondrocyte function, and anti-inflammation. Lorecivivint shows potential to modify structure and improve symptoms of knee OA.

A small-molecule inhibitor of the Wnt pathway (SM04690) as a potential disease modifying agent for the treatment of osteoarthritis of the knee.

OBJECTIVES: Osteoarthritis (OA) is a degenerative disease characterized by loss of cartilage and increased subchondral bone within synovial joints. Wnt signaling affects the pathogenesis of OA as this pathway modulates both the differentiation of osteoblasts and chondrocytes, and production of catabolic proteases. A novel small-molecule Wnt pathway inhibitor, SM04690, was evaluated in a series of in vitro and in vivo animal studies to determine its effects on chondrogenesis, cartilage protection and synovial-lined joint pathology. DESIGN: A high-throughput screen was performed using a cell-based reporter assay for Wnt pathway activity to develop a small molecule designated SM04690. Its properties were evaluated in bone-marrow-derived human mesenchymal stem cells (hMSCs) to assess chondrocyte differentiation and effects on cartilage catabolism by immunocytochemistry and gene expression, and glycosaminoglycan breakdown. In vivo effects of SM04690 on Wnt signaling, cartilage regeneration and protection were measured using biochemical and histopathological techniques in a rodent acute cruciate ligament tear and partial medial meniscectomy (ACLT + pMMx) OA model. RESULTS: SM04690 induced hMSC differentiation into mature, functional chondrocytes and decreased cartilage catabolic marker levels compared to vehicle. A single SM04690 intra-articular (IA) injection was efficacious in a rodent OA model, with increased cartilage thickness, evidence for cartilage regeneration, and protection from cartilage catabolism observed, resulting in significantly improved Osteoarthritis Research Society International (OARSI) histology scores and biomarkers, compared to vehicle. CONCLUSIONS: SM04690 induced chondrogenesis and appeared to inhibit joint destruction in a rat OA model, and is a candidate for a potential disease modifying therapy for OA.

Integration of reverse transcriptase loop-mediated isothermal amplification with an immunochromatographic strip on a centrifugal microdevice for influenza A virus identification.

A novel centrifugal microdevice which could perform reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) and immunochromatographic strip (ICS) based amplicon detection was demonstrated for simple and cost-effective influenza A virus identification. The proposed centrifugal microdevice consists of the sample and running buffer loading reservoirs, the RT-LAMP chamber, and the ICS for detecting gene expression. The entire process could be completed sequentially and automatically by simply controlling the rotation speed and by optimizing the microfluidic design. Monoplex and multiplex RT-LAMP reactions targeting H1 and/or M gene were executed at 66 °C for 40 min, and the resultant amplicons were successfully analysed on the ICS within 15 min. Influenza A H1N1 virus was subtyped by detecting H1 and M gene on the ICS even with 10 copies of viral RNAs. Highly specific and multiplex viral typing of the integrated RT-LAMP-ICS microdevice was also demonstrated. The combination of the rapid isothermal amplification with the simple colorimetric detection on a strip in a single centrifugal microdevice will provide an advanced genetic analysis platform in the field of on-site pathogen diagnostics.

Acetylation changes at lysine 5 of histone H4 associated with lytic gene promoters during reactivation of Kaposi's sarcoma-associated herpesvirus.

Kaposi's sarcoma-associated herpesvirus (KSHV) is a pathogenic agent of Kaposi's sarcoma, primary effusion lymphoma and multicentric Castleman's disease in humans. Similarly to other gammaherpesviruses such as Epstein-Barr virus (EBV) and herpesvirus saimiri (HVS), KSHV displays two alternative life cycles, latent and lytic one. The transactivation from latency to the lytic phase is the result of transcriptional changes in the KSHV genome caused by the replication and transcriptional activator (RTA). During KSHV reactivation, epigenetic modifications of histone protein on the viral genome occur, which regulate the transcriptional activation of a number of lytic genes. The reactivation of EBV from latency to lytic cycle, induced by an immediate-early Zta protein, was shown to be accompanied by acetylation of specific lysines in histone H4. Accordingly, we hypothesized that the RTA-induced transactivation of KSHV could also be accompanied by histone acetylation. To validate this hypothesis, we assayed alterations of acetyl-histone H4-lysine 5 (acH4K5) during the RTA-mediated KSHV reactivation. While the modified histone protein in a total cell lysate was not distinguished between control and RTA-expressed cells, upregulated acH4K5 was detected on several lytic gene promoter regions during KSHV reactivation. Our results clearly indicate that this epigenetic change is related to transcription of genes expressed in the lytic cycle of KSHV.

CHIP has a protective role against oxidative stress-induced cell death through specific regulation of endonuclease G.

Oxidative stress is implicated in carcinogenesis, aging, and neurodegenerative diseases. The E3 ligase C terminus of Hsc-70 interacting protein (CHIP) has a protective role against various stresses by targeting damaged proteins for proteasomal degradation, and thus maintains protein quality control. However, the detailed mechanism by which CHIP protects cells from oxidative stress has not been demonstrated. Here, we show that depletion of CHIP led to elevated Endonuclease G (EndoG) levels and enhanced cell death upon oxidative stress. In contrast, CHIP overexpression reduced EndoG levels, and resulted in reduced or no oxidative stress-induced cell death in cancer cells and primary rat cortical neurons. Under normal conditions Hsp70 mediated the interaction between EndoG and CHIP, downregulating EndoG levels in a Hsp70/proteasome-dependent manner. However, under oxidative stress Hsp70 no longer interacted with EndoG, and the stabilized EndoG translocated to the nucleus and degraded chromosomal DNA. Our data suggest that regulation of the level of EndoG by CHIP in normal conditions may determine the sensitivity to cell death upon oxidative stress. Indeed, injection of H2O2 into the rat brain markedly increased cell death in aged mice compared with young mice, which correlated with elevated levels of EndoG and concurrent downregulation of CHIP in aged mice. Taken together, our findings demonstrate a novel protective mechanism of CHIP against oxidative stress through regulation of EndoG, and provide an opportunity to modulate oxidative stress-induced cell death in cancer and aging.

A method of reducing background radiance for emissivity-compensated radiation thermometry of silicon wafers.

We studied the spectral and directional emissivities of silicon wafers using an optical polarization technique. Based on simulation and experimental results, we developed two radiation thermometry methods for silicon wafers: one is based on the polarized emissivity-invariant condition and the other is based on the relationship between the ratio of the p- and s-polarized radiance and the polarized emissivity. These methods can be performed at temperatures above 600 °C and over a wide wavelength range (0.9-4.8 µm), irrespective of the dielectric film thickness and the substrate resistivity, which depends on the dopant concentration. The temperature measurements were estimated to have expanded uncertainties (k = 2) of less than 5 °C. With a view to practically applying these methods, we investigated a method to reduce the intense background radiance produced by high-intensity heating lamps. We found that the background radiance can be greatly reduced by using a radiometer that is sensitive to wavelengths of 4.5 or 4.8 µm and suitable geometrical arrangements of a quartz plate. This opens up the possibility of using the two proposed radiation thermometry methods in practical applications.

Comparison of transnasal small-caliber vs. peroral conventional esophagogastroduodenoscopy for evaluating varices in unsedated cirrhotic patients.

BACKGROUND AND STUDY AIMS: We aimed to evaluate the accuracy of transnasal small-caliber esophagogastroduodenoscopy (TNSC-EGD) compared with peroral conventional EGD (POC-EGD) for evaluating varices in unsedated patients with liver cirrhosis. The success rate, safety, endoscopist satisfaction, and patient tolerability of TNSC-EGD were also addressed. PATIENTS AND METHODS: One hundred patients with liver cirrhosis participated in this randomized crossover trial, and 84 subjects completed both procedures. Of the 84 patients, 28 had marked bleeding diathesis (platelet count ≤ 50000/mm (3) and/or prothrombin time ≥ 1.7 INR). Endoscopists and patients answered questionnaires using a 100-mm visual analog scale about, respectively, their satisfaction and their tolerance of the procedure. RESULTS: The success rate of TNSC-EGD was comparable to that of POC-EGD (96% vs. 99%). Nasal mucosal hemorrhages induced by TNSC-EGD occurred in 5 patients (6%), but were easily controlled. Compared to the POC-EGD reference test, diagnostic accuracies of TNSC-EGD for detecting esophageal varices, gastric varices, and red color signs were 98%, 98%, and 96%, respectively. Concordance rates on grading esophageal varices and gastric varices were excellent at 93% (κ = 0.85) and 96% (κ = 0.87). Endoscopist satisfaction was not significantly different between TNSC-EGD and POC-EGD, whereas patient tolerance of TNSC-EGD was significantly greater than that of POC-EGD (79.0 ± 14.4 vs. 69.5 ± 16.1; P = 0.001). CONCLUSION: TNSC-EGD without sedation was found to be feasible, safe, and accurate for evaluating esophageal varices, gastric varices, and red color signs in patients with cirrhosis - even in those with marked bleeding diathesis. Furthermore, it was significantly better tolerated by patients, without altering endoscopist satisfaction. Our findings indicate that TNSC-EGD without sedation might be viewed as a potential alternative to POC-EGD for evaluation of varices.

Spinal cord injury subsequent to transcatheter arterial chemoembolization in patients with hepatocellular carcinoma.

Transcatheter arterial chemoembolization is one of the most common treatment modalities for hepatocellular carcinoma. Transcatheter arterial chemoembolization is considered to be a relatively safe procedure, but transcatheter arterial chemoembolization is associated with a number of disastrous complications. Among the ischaemic complications caused by transcatheter arterial chemoembolization, spinal cord injury is very rare, but can occur via the intercostal or lumbar arteries. We report two cases of extremely rare spinal cord injuries after transcatheter arterial chemoembolization in patients with hepatocellular carcinoma. The patients had sensory loss below the T9 or T10 dermatomes and paraparesis or paraplegia within 6-8h after transcatheter arterial chemoembolization. One patient sustained paraplegia until death 2 months after transcatheter arterial chemoembolization and the other patient recovered almost completely 2 months after transcatheter arterial chemoembolization.

Percutaneous aspiration thrombectomy for the treatment of acute lower extremity deep vein thrombosis: is thrombolysis needed?

AIM: To assess the technical feasibility and initial success of aspiration thrombectomy as a potential alternative to lytic therapy in initial endovascular management of acute lower extremity deep vein thrombosis (DVT). MATERIALS AND METHODS: From July 2004 to October 2007, a retrospective analysis of 27 patients (male:female 5:22; mean age 59 years) with acute iliofemoral or femoropopliteal DVT of less than 2 weeks was performed. All patients underwent sonography of the lower extremities, and 13 patients underwent computed tomography (CT) venography. All patients received an inferior vena cava (IVC) filter and were initially treated with aspiration thrombectomy using the pullback technique with or without basket thrombus fragmentation. If persistent stenotic portions (>50% luminal narrowing) were noted, balloon angioplasty or stent placement was performed. Successful recanalization was defined as successful restoration of antegrade flow in the treated vein with elimination of any underlying obstructive lesion. RESULTS: The mean procedure time was 65 min (range 40-100 min). Successful initial recanalization was achieved in 24 patients (88.9%) without complications. Urokinase was required for three patients (11.1%) due to a hard thrombus remaining in the iliac vein. Of the 27 patients, 23 had residual venous stenosis in the common iliac vein or external iliac vein. Therefore, balloon angioplasty (n=23) and stent placement (n=22) was performed. The remaining four patients were treated using only aspiration thrombectomy without angioplasty or stent placement. CONCLUSION: Aspiration thrombectomy without catheter-directed thrombolysis is a safe and effective treatment for acute DVT of the lower extremities, and minimizes the risk of haemorrhagic complications.

Diagnosis and treatment of obscure gastrointestinal bleeding using combined capsule endoscopy and double balloon endoscopy: 1-year follow-up study.

BACKGROUND AND STUDY AIMS: Several studies have shown the value of capsule endoscopy and double balloon endoscopy (DBE) in small-intestinal bleeding. The purpose of this study was to evaluate the impact of capsule endoscopy results on subsequent DBE examination, and the 1-year clinical outcome of this combined approach in patients with obscure gastrointestinal bleeding (OGIB). PATIENTS AND METHODS: A total of 45 consecutive patients with OGIB underwent capsule endoscopy. Patients with positive capsule endoscopy results underwent DBE for biopsy or therapy, and those with negative results underwent further assessment for possible diagnostic misses on capsule endoscopy. Tumors, ulcerations, and vascular lesions were considered as sources of bleeding. Diagnoses of OGIB lesions and clinical outcome were assessed 1 year after these examinations. RESULTS: Responsible lesions were found in 22 patients (49 %): 19 lesions in 18/45 patients (40 %) undergoing capsule endoscopy, and 18/36 patients (50 %) undergoing subsequent DBE. In all, 10 tumors, nine vascular lesions, and four ulcerations were found. In two patients, vascular lesions were only later diagnosed by conventional methods (4 %). Capsule endoscopy results guided our choice of the proper DBE model for successful therapeutic intervention in five patients. Re-bleeding rates were low during 1-year follow-up of the entire group (mean follow-up, 18.8 months): 5 % in cases with positive diagnoses on capsule endoscopy and/or DBE, and 12 % in negative cases. CONCLUSIONS: A combined approach using capsule endoscopy followed by DBE proves valuable in the diagnosis and treatment of patients with OGIB, leaves a low rate of undiagnosed bleeding sources, and has a good long-term outcome.

Effects of intracerebroventricular infusions of arginine vasopressin in sheep.

In sheep, neither the in vivo effect of vasopressin administered by a method other than systemic infusion nor the central effects on behavior from the perspective of stress regulation has been fully elucidated in an intact animal. We examined changes in behavioral, adrenocorticotropic, and autonomic nervous functions after intracerebroventricular infusions of arginine vasopressin (AVP) to elucidate its central role. Intracerebroventricular infusions of AVP (0, 0.12, 1.2 and 12 microg/500 microl/30 min) evoked a dose-related increase in plasma cortisol concentration. There were significant treatment-related effects on the total duration of sham-chewing (Friedman's test, X2=12.75, p=.0052), on the total duration of bar-biting (Friedman's test, X2=15.0, p=.0018), and on the total duration of rubbing (Friedman's test, X2=12.0, p=.0074). AVP 12 microg treatment induced a greater degree of sham-chewing and bar-biting than the other three treatments did (Nemenyi multiple comparisons: p<0.1). These findings indicate, together with our previous findings, that AVP has the same corticotropic potential as corticotropin-releasing hormone infused intracerebroventricularly in equal molar concentrations. Although the degree to which central stress signaling pathways are involved in these responses remains speculative, the relationships between stereotypies and central AVP are of particular interest.

Monocyte chemoattractant protein 1 and macrophage cyclooxygenase 2 expression in colonic adenoma.

BACKGROUND AND AIMS: Cyclooxygenase 2 (COX-2) expression in subepithelial macrophages of colorectal adenoma has been suggested as the first in a series of steps leading to colorectal tumorigenesis. We tested the hypothesis that chemokines released from human colorectal adenoma epithelium might be involved in COX-2 expression in macrophages of the lamina propria. METHODS: Endoscopic samples of sporadic colorectal adenomas were tested by enzyme linked immunosorbent assay for chemokines involved in macrophage chemotaxis. Localisation of adenoma macrophage chemoattractant protein 1 (MCP-1) and COX-2 were determined by immunohistochemistry. The effects of MCP-1, in the presence or absence of celecoxib, on COX-2 expression, and prostaglandin (PG) E(2) and vascular endothelial growth factor (VEGF) release, were examined in human macrophages isolated from peripheral blood. RESULTS: MCP-1 levels were markedly higher in adenoma with mild-moderate dysplasia (129.7 (19.9) pg/mg protein) and severe dysplasia (227.9 (35.4) pg/mg protein) than in normal colonic mucosa (55.8 (4.2) pg/mg protein). Other chemokine levels, macrophage inflammatory proteins (MIP)-1alpha and MIP-1beta, and the chemokine regulated on activation of normal T cell expressed and secreted (RANTES) did not vary significantly between adenoma and normal mucosa. MCP-1 levels in both adenoma and normal colonic mucosa increased significantly three hours after tissue cultivation in vitro. MCP-1 immunoreactivity was restricted to the adenoma epithelium, with no reactivity seen in adjacent normal epithelial cells. MCP-1 stimulated COX-2 expression and PGE(2) and VEGF release in human macrophages. Celecoxib, a selective COX-2 inhibitor, inhibited MCP-1-induced PGE(2) and VEGF release in macrophages. Addition of exogenous PGE(2) reversed this inhibitory effect on VEGF release, suggesting that MCP-1 in adenoma epithelial cells might be involved in COX-2 expression and subsequent macrophage activation. CONCLUSIONS: MCP-1 in colorectal adenoma epithelial cells might be involved in macrophage migration and COX-2 expression, leading to the subsequent development of colonic adenoma.

Spiral CT of the gastric carcinoma: staging and enhancement pattern.

The purpose of this study was to correlate the preoperative staging of gastric carcinoma using spiral computed tomography (CT) with pathologic staging and to correlate the enhancement pattern of advanced gastric carcinoma (AGC) on spiral CT with histological type. A total of 180 patients with gastric carcinomas confirmed at surgery underwent spiral CT. After surgery, pathologic findings were compared with CT findings. The detection rate for T1 tumors was 81.4% (57/70), and all T2-4 tumors were detected (110/110). In the T class, good correlation with pathology occurred in 47.8%. In the N class, good correlation with pathology occurred in 52.2%. The rate of understaging in the N class (31.7%) was higher than that of overstaging (16.1%) (P<.001). In AGC, the tumor mass showed delayed enhancement, regardless of Borrmann's type. By histological pattern, good and delayed enhancement was seen in 2/3 (66.7%) with signet ring cell type, but 4/5 (80%) with mucinous type were poorly enhanced (P=.019). Spiral CT for determining the preoperative staging of gastric carcinoma was not accurate. However, the enhancement pattern of AGC correlated with histological type.

Aggressive preoperative management and extended surgery for inflammatory pseudotumor involving the hepatic hilum in a child.

Inflammatory pseudotumors involving the hepatic hilum are rare. Only 14 cases have been reported (Table). Liver transplantation has been required when the pseudotumor has invaded extensively into the right and left lobes. (1,2) However, transplantation is associated with the lifelong use of immunosuppressants. This is particularly problematic in children. we report a case of a 6-year-old boy with an inflammatory pseudotumor extensively invading the hepatic hilum who was treated with aggressive surgical excision using the techniques devised for the treatment of hilar cholangiocarcinoma. (3)

Viral interferon regulatory factor 1 of Kaposi's sarcoma-associated herpesvirus binds to p53 and represses p53-dependent transcription and apoptosis.

Kaposi's sarcoma-associated herpesvirus (KSHV) is related to the development of Kaposi's sarcoma. Open reading frame K9 of KSHV encodes viral interferon regulatory factor 1 (vIRF1), which functions as a repressor of interferon- and IRF1-mediated signal transduction. In addition, vIRF1 acts as an oncogene to induce cellular transformation. Here we show that vIRF1 directly associates with the tumor suppressor p53 and represses its functions. The vIRF1 interaction domains of p53 are the DNA binding domain (amino acids [aa] 100 to 300) and the tetramerization domain (aa 300 to 393). p53 interacts with the central region (aa 152 to 360) of vIRF1. vIRF1 suppresses p53-dependent transcription and deregulates its apoptotic activity. These results suggest that vIRF1 may regulate cellular function by inhibiting p53.

Differential modulation of ACAT1 and ACAT2 transcription and activity by long chain free fatty acids in cultured cells.

Fatty acyl CoA and cholesterol are the substrates for cholesteryl ester synthesis by acyl coenzyme A:cholesterol acyltransferase (ACAT). Two ACAT genes have been identified; ACAT1 is expressed ubiquitously while ACAT2 is primarily expressed in intestine and liver. We tested effects of different free fatty acids (FFAs) on ACAT1 and ACAT2 expression and activity in HepG2 human hepatocytes and THP1 human macrophages. Incubation of oleic acid, arachidonic acid, or eicosapentaenoic acid, but not 25-hydroxycholesterol, induced ACAT1 mRNA levels 1.5--2-fold in HepG2, with no affect on ACAT2 mRNA. FFA had no affect on ACAT1 mRNA in THP1 cells. To determine if FFAs affect ACAT1 or ACAT2 posttranscriptionally, cells were labeled with [(3)H]cholesterol in the presence of the different FFAs for 1--5 h. Both HepG2 and THP1 cells showed the greatest cholesteryl ester production with oleic acid. This was also confirmed by the observation that more [(3)H]oleic acid incorporated into CE compared to [(3)H]eicosapentaenoic acid, even though there was no difference in the total uptake of these FFAs. In ACAT-deficient SRD4, CHO cells stably transfected with human ACAT1 or ACAT2, ACAT1 expressing cells showed a strong preference for oleic acid while ACAT2 expressing cells utilized unsaturated FFAs. Acyl CoA substrate specificity was further tested in microsomes isolated from these cells as well as HepG2 and THP1. THP1 and ACAT1 cells utilized oleoyl CoA preferentially. In contrast, HepG2 and ACAT2 microsomes utilized linolenoyl CoA as well. We conclude that FFAs increase ACAT1 mRNA levels in a cell specific manner, and furthermore that the ACAT reactions exhibit differential FFA utilization.

Radiologic anatomy of the rabbit liver on hepatic venography, arteriography, portography, and cholangiography.

UNLABELLED: Seo TS, Oh JH, Lee DH, et al. Radiologic anatomy of the rabbit liver on hepatic venography, arteriography, portography, and cholangiography. Invest Radiol 2001;36:186-192. RATIONALE AND OBJECTIVES: The radiologic anatomy of rabbit liver has received little study but is important in many experimental investigations. METHODS: Twenty-four rabbits were studied by using hepatic venograms, aortograms, hepatic arteriograms, cholangiograms, and portograms. RESULTS: In all cases, the right, middle, and left hepatic veins drained into the inferior vena cava just below the diaphragm, and the caudate lobe hepatic vein drained more inferiorly. The proper hepatic artery was a branch of the common hepatic artery in 96%. The first branch of the proper hepatic artery was the caudate lobe artery. The remaining main hepatic artery was divided into the right and left hepatic arteries. The left hepatic artery was further divided into the medial and lateral segmental branches in 95%. The anatomy of the portal vein or bile duct was the same as the hepatic artery in 100% of cases. CONCLUSIONS: Knowledge of the normal patterns and variations of the vessels and bile duct will be helpful for experiments of the rabbit liver in future studies.

Two-dimensional alterations of myenteric plexus in jejunoileal atresia.

PURPOSE: The purpose of this report is to investigate changes in the myenteric plexus associated with the dilated proximal segment of jejunoileal atresia (JA). Two-dimensional morphologic changes in the myenteric nerve plexuses were investigated using whole-mount preparation. METHODS: Proximal (P) and distal (D) intestinal segments from 7 cases with JA and control (C) segments from 5 postmortem neonates were investigated. The circumference of the jejunoileal segments was measured after fixation. Antibodies for protein gene product 9.5 and neurofilament protein were used in whole-mount preparation. The sizes of neural networks were calculated by measuring the longest circular distance in a neural network (x) and the longest longitudinal distance (y), and the width of the internodal strands (i) with a videomicrometer. RESULTS: Median circumference of the segments was 8.5 in P, 2.0 in D, and 2.0 cm in C. The neural networks in P were expanded longitudinally as well as circularly (x = 817.10 microm, y = 561.26, i = 31.04: median) while comparing them to those in D (x = 431.40 microm y = 288.07, i = 26.05) or C (x = 285.03 microm y = 372.20, c = 1113.57, i = 32.39). The nerve plexus was less expanded than the intestinal wall. CONCLUSIONS: Proximal intestinal segments showed a restructuring that resulted in mild hypoplasia of the enteric nerve plexuses in the proximal segments. The less expansion of the myenteric nerve plexus seen in the proximal bowel in infants with JA suggests a histologic basis for the efficacy of tapering or plication of the dilated bowel.

Long-term follow-up of the clinical efficacy of chemotherapy for acute myeloid leukemia at a single institute.

A retrospective study was performed on 125 patients with de-novo acute myeloid leukemia (AML) who had received first remission induction therapy at Fukui Medical University Hospital in the 16 years between 1983 and 1998. For remission induction therapies, patients in the 1980s received mainly behenoylcytarabine (BHAC), 6-mercaptopurine (6-MP), and prednisolone (PSL), plus aclarubicin (ACR) or daunorubicin (DNR). Patients in the 1990s received mainly BHAC, 6-MP, and etoposide (VP-16) plus DNR or mitoxantrone (MIT) or idarubicin (IDA). Patients with hypoplastic bone marrow received low-dose cytarabine (Ara-C) therapy or cytarabine ocfosfate (SPAC). Since 1992, patients with French-American-British disease classification of M3 have received all-trans retinoic acid (ATRA) (+/-chemotherapy). In the 1990s, more intensified postremission therapy was performed compared with that done in the 1980s. The complete remission (CR) rate of all patients was 58%. Predicted 6-year overall survival (OS) and disease-free survival (DFS) rates in the CR patients were 22% and 28%, respectively. Multivariate analysis showed age and leukocyte counts as significant prognostic factors regarding CR, OS, and DFS rates. The CR and OS rates in the 1990s were improved significantly from those in the 1980s, at 69% versus 48% (P = 0.016), and 32% versus 15% (P = 0.0014), respectively. The early death rate, within 30 days, was decreased from 26% in the 1980s to 9% in the 1990s (P = 0.013). This decrease was thought to be the main cause of the high CR rate in the 1990s. However, DFS was not significantly improved. It is necessary to establish more effective postremission therapies in order to reduce the relapse rate and improve the prognosis.

[Audiological and equilibrium study of perilymphatic fistulas].

We clinically analyzed 15 cases of perilymphatic fistulas--11 caused by barotraumas and 4 idiopathic--identified by surgery between March 1995 and March 1999 at the Hyogo College of Medicine and affiliated hospitals. Subjects were 11 men and 4 women (aged 14 to 79 years (mean: 46.7 years)). All showed hearing loss in audiography and 12 cases reported tinnitus--stream-like in 5 and poping in 4. Dysequilibrium was seen in 9 cases. Perilymph leakage was detected intraoperatively from the oval window in 9, from the round window in 4, and from both windows in 1, while another had leakage from the fissura ante fenestram. After surgery, hearing level improved by over 10 dB in 9 of the 11 cases operated on within 14 days after onset. Hearing did not improve in 3 of 4 operated on later. Vertigo disappeared after surgery. Dizziness tended to persist in those having canal paresis or paralytic nystagmus before surgery. We suggest that patients with progressive hearing loss should be operated on as soon as possible and that patients with dysequilibrium or without response to conservative treatment undergo surgery within 14 days of onset.