Optical mapping of pontine chemosensitive regions of neonatal rat

We analyzed the neuronal response to hypercapnic acidosis, using an optical recording technique with a fluorescent voltage-sensitive dye (di-4-ANEPPS), in pontine slice preparations of neonatal rats, containing the locus coeruleus (LC), which has been electrophysiologically demonstrated to be chemosensitive. The dye-stained preparation was continuously superfused with artificial cerebrospinal fluid. Epifluorescence of the slice was detected using a high-sensitivity optical recording system. Changes in the intensity of fluorescence were serially analyzed while switching artificial cerebrospinal fluid from control to hypercapnic acidosis, or vice versa. The optical recording method revealed that the LC, as reported in previous studies, reversibly showed a depolarizing response to hypercapnic acidosis in 56% of the examined preparations. The A5 area (56%) also exhibited a reversible, depolarizing response to hypercapnic acidosis. The response was preserved under conditions in which chemical synaptic transmission was blocked by low Ca(2+)-high Mg(2+) solution. These results suggest that the optical recording method is applicable to identification of potentially chemosensitive areas, which deserve further electrophysiological analysis, and that the A5 area could be chemosensitive.     

Premature regression of the leg apical ectodermal ridge in the Japanese chick wingless mutant

The autosomal dominant Japanese wingless mutant has varying degrees of wing and leg truncations. The wing defects range from complete loss to negligible defects, whereas leg abnormalities are usually restricted to loss of the phalanges. Further analyses of the mutant focusing on the leg, which has been relatively uncharacterized, were performed. The expression pattern of Fgf8, a marker gene for the apical ectodermal ridge (AER) that controls outgrowth of the limbs, revealed premature regression at stage 28. Electron microscopy study showed abnormalities in the basement membrane all through the AER in the same stage. In the mutant, cell death was observed in the mesenchyme underlying AER between stages 31 and 32, although in the wild-type leg, AER regression and cell death occurred almost simultaneously at stages 33–34. To know if the cell death and cessation of the outgrowth are common mechanisms of wild-type and the mutant, we removed the AER in wild-type embryos at stage 28 and followed the fate of the limb. This also resulted in premature cell death 48 h after AER removal (equivalent to stage 32) and limb truncations similar to those observed in mutant limbs. To confirm whether either AER or underlying mesenchyme is responsible for the truncation, transplantation of the AER between the wild-type and the mutant was performed. This revealed that AER is the defective tissue in this mutant.     

Relationship between cochlear implant outcome and the diameter of the cochlear nerve depicted on MRI

This study aimed to evaluate the relationship between the diameters of the auditory and eighth cranial nerves and improvements in post-implant performance. Twenty prelingually deafened children (aged from 2.0 to 6.0 years) who received the Nucleus 24 cochlear implant participated in this study. All subjects had used their implant for at least 1 year after device connection. The diameters of cochlear and eighth cranial nerves were retrospectively measured on preoperative T2-weighted axial magnetic resonance image (MRI). In 17 of 20 subjects, the cochlear and eighth cranial nerves could be identified on MRI. The mean diameter of the cochlear and eighth cranial nerves were 0.9 +/- 0.2 mm and 1.2 +/- 0.3 mm, respectively. In the remaining three subjects, the cochlear and eighth cranial nerves could not be identified on MRI. These three subjects had significantly lower scores in the Infant-Toddler-Meaningful Auditory Integration Scale (IT-MAIS) than the other 17 subjects at 12 months post-implant. There was no significant correlation among the maximal diameters of the nerves and age, ECAP thresholds and IT-MAIS scores. A sufficient outcome from cochlear implantation can be expected when cochlear and eighth cranial nerves are depicted on MRI, regardless of the nerve diameters.     

Rapidly prototyped temporal bone model for otological education

The anatomy of the temporal bone is extremely complicated. If a three-dimensional model could be simulated, it would greatly contribute to the stereoscopic understanding of anatomy and surgery. A simulated three-dimensional model of a human temporal bone was prototyped using the selective laser sintering method. The model could be shaved using a surgical drill in the same way as in real surgery. A magnified model was particularly useful for the instruction of anatomy and surgery. When a translucent area was selected, a bony labyrinth could be created together with an internal auditory meatus and facial nerve, which also contributed to the easy understanding of the inner ear structure. The three-dimensional prototyped model using selective laser sintering serves as a good educational material for middle ear anatomy and surgery.     

Imaging of acute pancreatitis--the role of US, CT and MRI

Imaging of the pancreas with US and CT plays a major role in the diagnosis and evaluations of patients with acute pancreatitis. In severe acute pancreatitis, dynamic CT is essential to assess the severity of the disease and evaluate the complications. MRI is comparable to CT in its capacity to provide precise information about the severity of acute pancreatitis. MRI has some advantages over CT in being free from ionizing radiation and lower toxity of gadolinium which is used for contrast medium intravenously. The disadvantages of MRI, however, include cost, longer exam duration, limited availability and the practical difficulties of scanning a patient with ICU equipment.     

Lymph node metastasis of gastric cancer: comparison of Union International Contra Cancer and Japanese systems

BACKGROUND: The pN classification of gastric cancer (GC) in the Japanese system (Japanese Gastric Cancer Association; JGCA) is based on the site and distance of metastatic nodes from the primary tumour. Union International Contra Cancer (UICC) has recently proposed a classification system based on the number of nodes involved (TNM-1997). The aim of the present study is to assess which classification system is more suitable for providing a prognosis in advanced GC with lymph node metastasis. METHODS: A total of 224 patients who underwent curative gastrectomy (R0: UICC-TNM and Resection A and B: JGCA) and D2 lymphadenectomy between 1990 and 1999, and diagnosed as pT2, pT3 and pT4 GC were enrolled. Patients were followed until the end of 2002. The disease-free survival rates of patients were compared between the two-stage systems (UICC-TNM and JGCA). RESULTS: Using the JGCA system, there was a significant difference between the two survival curves (pN0 and pN1, P = 0.025; pN1 and pN2, P < 0.001; pN2 and pN3, P = 0.031), but the 5-year survival rate of 27 pN2 patients (32.7%) was not significantly different from that of 14 pN3 patients (34.3%, P = 0.994) using the UICC-TNM. In 47 patients with JGCA pN2, the 5-year survival rate of 18 patients with UICC-TNM pN1 (42.9%) was not significantly different from that of 18 patients with UICC-TNM pN2 (25.2%, P = 0.422) or from that of 11 patients with UICC-TNM pN3 (24.2%; P = 0.383). CONCLUSIONS: The JGCA system is more suitable for estimating the prognosis of Japanese patients with advanced GC than the UICC-TNM.     

Osteopontin is a negative regulator of proliferation and differentiation in MC3T3-E1 pre-osteoblastic cells

Osteopontin (OPN) is an important mediator of bone remodeling. However, the role of OPN in the process of bone formation is not fully understood. In previous studies, we have shown that MC3T3-E1 pre-osteoblastic cells at higher passage number exhibited weakened osteogenic capacity and elevated OPN mRNA expression. In this work, we investigated the role of OPN on proliferation and differentiation of low-passage MC3T3-E1 cells by studying stable cell lines overexpressing either OPN mRNA or its antisense RNA. Overexpression was verified by both Northern and Western blot analyses. Overexpression of OPN markedly inhibited proliferation as determined by daily cell counts, while overexpression of antisense RNA stimulated cellular proliferation. We also examined the effect of OPN level on BMP-2-induced alkaline phosphatase activity. Overexpression of OPN inhibited BMP-2 responsiveness while overexpression of antisense RNA enhanced the effect of BMP-2 on alkaline phosphatase activity. Increased OPN expression also caused decreases in expression of osteocalcin and bone sialoproteins while a reduction of OPN level caused the opposite. Furthermore, endogenous OPN expression in response to BMP-2 exhibited a biphasic pattern, that is, it was initially inhibited and then enhanced by the treatment of BMP-2, indicating that OPN might function as a negative feedback regulator for osteoblastic differentiation. Finally, overexpression of OPN inhibited mineral deposition. In contrast, overexpression of antisense RNA enhanced mineral deposition. These results indicate that OPN is a negative regulator of proliferation and differentiation in MC3T3-E1 cells.