Morphological characteristics of the vallate papillae of the one-humped camel (Camelus dromedarius).

In this study, the morphology of the vallate papillae of camel was investigated using gross, light and scanning electron microscopy as well as immunohistochemistry. Vallate papillae were arranged along an identical line on each side of the lingual torus and revealed remarkable individual differences. However, each papilla - round or flat, small or large, single or paired - was surrounded by a prominent groove and an annular pad. Based on our findings, postnatal development and formation of new papillae occur in camel. Microscopically, taste buds were constantly observed along the medial wall epithelium, and in the papillary wall epithelium on both sides of the secondary groove apparently separating the vallate papillae. In addition, an aggregation of taste buds was occasionally observed at the bottom of the lateral wall epithelium. Using SEM, we observed several pits and microplicae on the surface of papillae as well as distinct taste pores on the peripheral parts of the dorsal surface. We demonstrated immunoreactivity of α-gustducin only in mature taste buds. We conclude that the morphological features and microstructure of vallate papillae are a characteristic feature in camel compared to other ruminants. These features might have evolved to assist the camel in the manipulation and tasting of thin organic stiff plants that grow in its environment and therefore might have related to the feeding habits of the animal.

Skewed TGFß/Smad signalling pathway in T cells in patients with Behçet's disease.

OBJECTIVES: Behçet's disease (BD) is a multi-systemic inflammatory disease, characterised by recurrent oral aphthosis, genital ulcers, skin lesions and uveitis. We have reported excessive Th1 cell activity in patients with BD. More recently, Th17 cells were suggested to associate with several autoimmune diseases. This study was designed to investigate the role of Th17 related cytokines and signalling molecules in patients with BD. METHODS: We examined mRNA expressions of Th1 and Th17 related cytokines and related signalling molecules in PBMC of 12 patients with BD and 14 normal controls (NC) using quantitative RT-PCR. We studied expressions of the Th17 related cytokines in other four BD patients' skin lesions by immunofluorescence. RESULTS: Major Th17 related cytokines were not detected in unstimulated PBMC in patients with BD. After stimulation, mRNA expressions of TGFß receptor type 1, IL-12 receptor ß2 and suppressor of cytokine signalling protein (SOCS) 1 on PBMC were significantly enhanced in patients with BD, as compared with NC (p<0.05). mRNA expression of RORC, a key transcription factor for Th17 cell differentiation, was comparable between BD and NC. CD4+ T cells infiltrating into BD skin lesion expressed TGFß1 much more than those infiltrating into non-Behçet's disease erythema nodosum. CONCLUSIONS: These findings suggest that TGFß/Smad signalling pathway of T cells is overactive in patients with BD.

Excessive CD4+ T cells co-expressing interleukin-17 and interferon-γ in patients with Behçet's disease.

Excessive T helper type 1 (Th1) cell activity has been reported in Behçet's disease (BD). Recently, association of Th17 cells with certain autoimmune diseases was reported, and we thus investigated circulating Th17 cells in BD. CD4(+) CD45RO(-) (naive) T cells were cultured with Th0-, Th1-, Th2- and Th17-related cytokines and antibodies, and their mRNA was studied by real-time polymerase chain reaction (PCR). When naive CD4(+) T cells were cultured with Th1- and Th17-related cytokines, interferon (IFN)-γ mRNA and interleukin (IL)-17 mRNA were up-regulated, respectively, in BD patients. Naive CD4(+) T cells cultured in a Th17 cell-inducing condition expressed IL-23 receptor (IL-23R) mRNA excessively. IL-17 mRNA expression was induced only when naive CD4(+) T cells were cultured in the presence of IL-23. CD4(+) T cells cultured with Th17 cytokines expressed excessive RAR-related orphan receptor C (RORC) mRNA. Using intracellular cytokine staining, we found that CD45RO(+) (memory) CD4(+) T cells producing IL-17 and IFN-γ simultaneously were increased significantly. Memory CD4(+) T cells producing IFN-γ but not IL-17 decreased profoundly in BD patients. CD4(+) T cells producing IL-17 and IFN-γ simultaneously were found in BD skin lesions. Collectively, we found excessive CD4(+) T cells producing IL-17 and IFN-γ (Th1/Th17) cells in patients with BD, and possible involvement of IL-23/IL-23R pathway for the appearance of excessive Th1/Th17 cells.

Early CT signs in patients with acute middle cerebral artery occlusion: incidence of contrast staining and haemorrhagic transformations after intra-arterial reperfusion therapy.

AIM: To investigate how often early computed tomography (CT) signs are associated with blood-brain barrier (BBB) disruption and result in haemorrhagic transformations. MATERIALS AND METHODS: Serial CT findings were prospectively evaluated in 61 patients with acute middle cerebral artery (MCA) occlusion who underwent initial CT examination within 3h of stroke onset and who were treated with intra-arterial reperfusion therapy within 6h of stroke onset. Obscuration of the margin of the lentiform nucleus and loss of the insular ribbon were evaluated as early CT signs in the deep MCA territories. Cortical effacement was also evaluated. BBB disruption was defined as contrast medium staining in post-therapeutic CT just after reperfusion therapy. The relationship between pre-therapeutic early CT signs and post-therapeutic contrast staining or haemorrhagic transformations was investigated. RESULTS: The frequency of early CT signs in the deep MCA territories was significantly higher than that of cortical effacement (68.9 versus 27.9%). There were significant correlations between pre-therapeutic early CT signs and post-therapeutic contrast staining in both the deep and superficial MCA territories. Compared with early CT signs in the deep MCA territories, cortical effacement had a significantly higher incidence of post-therapeutic contrast staining (54.8 versus 82.4%). Although not statistically significant, cortical effacement tended to develop into haemorrhagic transformations. There was no correlation between early CT signs in the deep MCA territories and haemorrhagic transformations. CONCLUSION: Cortical effacement may be an advanced CT sign with BBB disruption and potential risk for haemorrhagic transformations. The presence of early CT signs in the deep MCA territories may not be a contraindication of reperfusion therapy.

Spontaneous regeneration of the corticospinal tract after transection in young rats: a key role of reactive astrocytes in making favorable and unfavorable conditions for regeneration.

We demonstrated the occurrence of marked regeneration of the corticospinal tract (CST) after a single transection and failure of regeneration after a repeated transection in young rats. To provide convincing evidence for the complete transection and regeneration we used retrograde neuronal double labeling. Double-labeled neurons that took up the first tracer from the transection site and the second tracer from the injection site caudal to the transection site were observed in the sensorimotor cortex. The anterograde tracing method revealed various patterns of regeneration. In the most successful cases the vast majority of regenerated fibers descended in the normal tract and terminated normally whereas a trace amount of fibers coursed aberrantly. In the less successful cases fibers descended partly normally and partly aberrantly or totally aberrantly. To clarify the role of astrocytes in determining the success or failure of regeneration we compared expression of glial fibrillary acidic protein (GFAP), vimentin and neurofilament (NF) immunoreactivity (IR) in the lesion between single and repeated transections. In either transection, astrocytes disappeared from the CST near the lesion site as early as 3 h after lesioning. However, by 24 h after a single transection, immature astrocytes coexpressing GFAP- and vimentin-IR appeared in the former astrocyte-free area and NF-positive axons crossed the lesion. By contrast, after a repeated transection the astrocyte-free area spread and NF-positive axons never crossed the lesion. It appears likely that the major sign, and possibly cause of failure of regeneration is the prolonged disappearance of astrocytes in the lesioned tract area.

Reperfusion Therapy for Acute Middle Cerebral Artery Trunk Occlusion. Direct Percutaneous Transluminal Angioplasty Versus Intra-arterial Thrombolysis.

SUMMARY: The purpose of this study was to test the hypothesis that direct percutaneous transluminal angioplasty (PTA) might reduce the incidence of haemorrhagic complications and might improve recanalization rate and clinical outcome as compared with intra-arterial (IA) thrombolysis in patients with acute middle cerebral artery (MCA) trunk occlusion. A total of 70 patients with acute MCA trunk occlusion were treated with IA reperfusion therapy. Thirty-six patients were treated with IA thrombolysis alone. In the other 34 patients, direct PTA was selected as the first choice of the treatment and subsequent thrombolysis was added if necessary for distal embolization. The modified Rankin scale (mRS) was used to assess clinical outcome at 90 days. As compared with IA thrombolysis, direct PTA provided significant increase in the rates of partial or complete recanalization (63.9 vs 91.2%, p < 0.01) and decrease in the incidence of large parenchymal hematoma with neurological deterioration (19.4% vs 2.9%, p=0.03). Despite such favorable effects, direct PTA did not improve the rate of a favorable outcome (mRS score 0 or 1, 41.7% for the IA thrombolysis group vs 52.9% for the PTA group, p=0.48). However, outcome classified in terms of independence (mRS score

Correlation between Haemorrhagic Complications and CT Findings before and after Intra-arterial Reperfusion Therapy for Acute Middle Cerebral Artery Occlusion.

SUMMARY: The purpose of this study was to investigate whether haemorrhagic complications can be predicted by evaluating CT findings before and after intra-arterial reperfusion therapy for acute middle cerebral artery (MCA) occlusion. Pretherapeutic early CT signs within three hours after onset and post-therapeutic contrast extravasation were evaluated in 61 patients treated within six hours after onset. Early CT signs were evaluated in the deep (obscuration of the margin of the lentiform nucleus and loss of the insular ribbon) and superficial MCA territories (cortical effacement). Haemorrhagic transformations were seen in 39.3% of patients, 54.2% of them had both pre-therapeutic early CT signs and post-therapeutic contrast extravasation. Obscuration of the entire lentiform nucleus and the presence of contrast extravasation were reliable predictors for haemorrhagic transformations, and cortical effacement had also a tendency to be associated with haemorrhage. Absence of early CT signs did not always result in the absence of haemorrhagic transformations and 37.5% of haemorrhage occurred in the presumed normal area without early CT signs. On the other hand, absence of post-therapeutic contrast extravasation was a reliable negative predictor for intraparenchymal haematoma.

Aberrant expression of Fas ligand on anti-DNA autoantibody secreting B lymphocytes in patients with systemic lupus erythematosus: "immune privilege"-like state of the autoreactive B cells.

BACKGROUND: Fas/Fas ligand (FasL) system has been assigned a pivotal role in the development and maintenance of peripheral tolerance, and mice with defects in their Fas/FasL system develop lupus-like symptoms. In this study we examined FasL expression of peripheral blood lymphocytes in patients with systemic lupus erythematosus (SLE). METHODS: We assessed FasL mRNA and protein expression by reverse transcription (RT)-PCR and immunoblotting and immunocytochemical staining, respectively, in patients with SLE. Anti-DNA antibody secreting B cells were purified using biotin labeled DNA and streptavidin-bead. RESULTS: Expression of FasL protein was not or very weakly detected in freshly isolated PBMC in normal individuals. In contrast, freshly isolated SLE PBMC exhibited the enhanced expression of FasL protein without in vitro stimulation. Not only purified T cells but also purified B cells expressed FasL on their cell surface spontaneously. In addition, freshly isolated anti-DNA autoantibody secreting B cells express FasL without in vitro stimulation. CONCLUSION: The results suggest that autoreactive B lymphocytes which aberrantly express FasL may kill Fas+ immunoregulatory T lymphocytes. Thus aberrantly expressed FasL may facilitate escape of the autoreactive B cells from the immune tolerance system, and may contribute to the sustained secretion of autoantibodies in patients with SLE.

Temporal and spatial distribution of Fos protein in the lumbar spinal dorsal horn neurons in the rat with chronic constriction injury to the sciatic nerve.

The temporal and spatial expression pattern of Fos protein in spinal dorsal horn neurons was examined by immunohistochemistry in rats with chronic constriction injury (CCI) to the sciatic nerve. In normal animals, a few Fos-immunoreactive (-IR) neurons were detected in the dorsal horn of the lumbar spinal cord. Following induction of CCI, a very large number of Fos-IR neurons appeared in the spinal dorsal horn, but a significant number of Fos-IR neurons were also observed in the contralateral dorsal horn where primary afferents of the injured sciatic nerve rarely project. Sham-operated animals also had a significant number of Fos-IR neurons in the dorsal horn bilaterally. The number of Fos-IR neurons reached its maximal level 1 day following placement of the ligatures (PO 1d). The ratio of the number of Fos-IR neurons in the ipsilateral dorsal horn to the contralateral dorsal horn, however, had its peak level 3 days following CCI (3.1-fold increase compared to the contralateral dorsal horn). The number of Fos-IR neurons in the dorsal horn gradually decreased, but increased again around PO 15d. On PO 30d, the number of Fos-IR neurons decreased and became comparable to that in normal animals. The present results indicate that the induction of Fos-IR neurons in the dorsal horn caused by CCI is biphasic and reaches its maximal level on PO 3d, near the time of hyperalgesia onset.

Serial changes in signal intensities of the adjacent discs on T2-weighted sagittal images after surgical treatment of cervical spondylosis: anterior interbody fusion versus expansive laminoplasty.

BACKGROUND: There have been many reports about newly developed degenerative changes in the adjacent segments after anterior interbody fusion. It is a controversial issue whether the adjacent-segment disease in patients treated by anterior interbody fusion is the result of progressive cervical spondylosis at the adjacent levels or is caused by the arthrodesis. The aim of this study is to clarify the difference in postoperative effect on the adjacent segments between anterior interbody fusion and expansive laminoplasty. METHOD: This study included 14 patients who underwent pre- and postoperative MR images at 6 and 12 months. Seven patients underwent cervical interbody fusion and the other 7 patients underwent expansive laminoplasty. Disc degeneration was evaluated semiquantitatively by calculating the degenerative index (DI) that is a ratio of the intensity in the disc to that in the upper cervical cord. FINDINGS: In the anterior interbody fusion group, the adjacent disc intensities decreased within 12 months (F = 20.42; P < 0.01). The pre-operative mean DI was 0.59 +/- 0.16. The post-operative mean DIs were 0.56 +/- 0.16 at 6 months and 0.47 +/- 0.16 at 12 months. In the expansive laminoplasty group, the signal intensities of both the adjacent discs and the discs within the range of laminoplasty had no serial changes during the same period (F = 2.67; P = 0.09 and F = 0.15; P = 0.87 respectively). INTERPRETATION: Anterior interbody fusion had a significant influence on the adjacent discs even as soon as 12 months after surgery, but laminoplasty had no influence on them during the same period.

Parenchymal hyperdensity on computed tomography after intra-arterial reperfusion therapy for acute middle cerebral artery occlusion: incidence and clinical significance.

BACKGROUND AND PURPOSE: The purpose of the present study was to assess the incidence and clinical significance of the intraparenchymal hyperdense areas on the posttherapeutic CT scan just after intra-arterial reperfusion therapy. METHODS: Seventy-seven patients with acute middle cerebral artery occlusion were studied prospectively with post-therapeutic CT. Hyperdense areas were classified into three groups: those in the lentiform nucleus, insular cortex and cerebral cortex. We investigated the incidence of hyperdense areas and hemorrhagic transformations and assessed whether location of hyperdense areas may play a role in the incidence of hemorrhagic transformations. We also evaluated correlation between early CT signs and hyperdense areas. RESULTS: Forty-five hyperdense areas were seen in 37 of the 77 patients (48.1%): 19 of the 45 (42.2%) were confirmed to be hematomas themselves, 6 (13.4%) showed later conversion to petechial hemorrhages, and 20 (44.4%) showed rapid disappearance without hemorrhagic transformations. Eleven of the 37 patients (29.7%) had neurological worsening due to massive hematoma (symptomatic hemorrhage), whereas none of the 40 patients without hyperdense areas had symptomatic hemorrhage. The incidence of hemorrhage among hyperdense areas was significantly lower in the insular cortex than in the other 2 regions (P<0.01). On the other hand, hyperdense areas in the lentiform nucleus had a significantly higher incidence of neurological worsening (P<0.05). There was a significant correlation between early CT signs and hyperdense areas (P<0.0001). CONCLUSIONS: The presence of hyperdense areas was a significant risk factor for severe hemorrhagic transformations, although only 29.7% of patients with hyperdense areas had symptomatic hemorrhage. On the contrary, the absence of hyperdense areas was a reliable negative predictor for symptomatic hemorrhage.

Temporal and spatial distribution of Fos protein in the parabrachial nucleus neurons during experimental tooth movement of the rat molar.

The present study was undertaken to reveal spatio-temporal changes in the distribution of Fos-like immunoreactive (-IR) neurons in the parabrachial nucleus (PBN), one of the important relay nuclei for processing autonomic and somatosensory information from the oro-facial regions, following the induction of experimental tooth movement in rat upper molars. The experimental tooth movement was induced by the insertion of elastic rubber between the first and second upper molars. In normal animals, the PBN contained a smaller number of Fos-IR neurons. Following experimental tooth movement, the Fos-IR neurons increased in number significantly on both the ipsilateral and contralateral PBN, reaching a maximum at 4 h (about 10 times that of normal animals), and then decreased gradually. However, a significant number of Fos-IR neurons remained at 24 h post-operation. Remarkable side-by-side differences in the number of Fos-IR neurons were recognized at 1 to 4 h following the experimental tooth movement. Their number returned to normal (basal) levels at 5 days post. All subnuclei of PBN showed similar temporal changes in the number of Fos-IR neurons, this being particularly apparent in lateral PBN. Administrations of morphine (3 and 10 mg/kg, i.p.) drastically reduced the induction of Fos-IR neurons in all subnuclei of both the ipsilateral and contralateral PBN in a dose-dependent manner, and its effect was antagonized by pretreatment with naloxone (2 mg/kg, i.p.). The reduction of Fos-IR neurons by morphine pretreatment suggests that the appearance of Fos-IR neurons in the PBN may be partly due to the noxious stimulation and/or stress arising from tooth movement. The bilateral expression of Fos-IR neurons in the PBN indicates that the experimental tooth movement causes the activation of PBN neurons for the processing of somatosensory as well as autonomic information. The prolonged expression of Fos-IR neurons in all the subnuclei of bilateral PBN reflects clinical features of the transient discomfort and/or abnormal sensations, which many patients often complain about during orthodontic treatment.

Reduced expression of hepatocyte growth factor activator inhibitor type-2/placental bikunin (HAI-2/PB) in human glioblastomas: implication for anti-invasive role of HAI-2/PB in glioblastoma cells.

Hepatocyte growth factor activator inhibitor type-2/placental bikunin (HAI-2/PB) is a serine proteinase inhibitor that contains 2 Kunitz-domains and a presumed transmembrane domain. It has broad inhibitory spectra against various serine proteinases showing potent inhibitory activities not only to hepatocyte growth factor activator but also to plasmin, trypsin and kallikreins. In this study, we investigated the expression of HAI-2/PB in human gliomas in vivo and the effects of HAI-2/PB on the fibrinolytic and invasive capabilities of human glioblastoma cells in vitro. With RNA blot analysis, HAI-2/PB mRNA was expressed in normal brain and in low-grade astrocytomas, but was hardly detectable in anaplastic astrocytomas and glioblastomas, indicating that its expression levels were inversely correlated with the histological grade of human gliomas. To further explore the possible role of HAI-2/PB in glioma progression, cultured human glioblastoma cell lines (U251 and YKG-1) were transiently transfected with an expression vector harboring human HAI-2/PB cDNA. Subsequent analysis indicated that the expression of HAI-2/PB suppressed the fibrinolytic activities of both glioblastoma cell lines. Moreover, HAI-2/PB inhibited Matrigel invasion of U251 and YKG-1 cells by 30% and 64%, respectively. This anti-invasive effect appeared to be mediated primarily by the inhibitory activity of HAI-2/PB against the serine proteinase-dependent matrix degradation. These findings suggest that the reduced expression of HAI-2/PB is possibly involved in the progression of human gliomas.

Development and maturation of taste buds of the palatal epithelium of the rat: histological and immunohistochemical study.

Palatal taste buds are intriguing partners in the mediation of taste behavior and their spatial distribution is functionally important for suckling behavior, especially in the neonatal life. Their prenatal development has not been previously elucidated in the rat, and the onset of their maturation remains rather controversial. We delineated the development and frequency distribution of the taste buds as well as the immunohistochemical expression of alpha-gustducin, a G protein closely related to the transduction of taste stimuli, in the nasoincisor papilla (NIP) and soft palate (SP) from the embryonic day 17 (E17) till the postnatal day 70 (PN70). The main findings in the present study were the development of a substantial number of taste pores in the SP of fetal rats (60.3 +/- 1.7 out of 122.8 +/- 5.5; mean +/- SD/animal at E19) and NIP of neonatal rats (9.8 +/- 1.0 out of 44.8 +/- 2.2 at PN4). alpha-gustducin-like immunoreactivity (-LI) was not expressed in the pored taste buds of either prenatal or newborn rats. The earliest expression of alpha-gustducin-LI was demonstrated at PN1 in the SP (1.5 +/- 0.5 cells/taste bud; mean +/- SD) and at PN4 in the NIP (1.4 +/- 0.5). By age the total counts of pored taste buds continuously increased and their morphological features became quite discernible. They became pear in shape, characterized by distinct pores, long subporal space, and longitudinally oriented cells. Around the second week, a remarkable transient decrease in the total number of taste buds was recorded in the oral epithelium of NIP and SP, which might be correlated with the changes of ingestive behaviors. The total counts of cells showing alpha-gustducin-LI per taste bud gradually increased till the end of our investigation (14.1 +/- 2.7 in NIP and 12.4 +/- 2.5 in SP at PN70). We conclude that substantial development of taste buds began prenatally in the SP, whereas most developed entirely postnatal in the NIP. The present study provides evidence that the existence of a taste pore which is considered an important criterion for the morphological maturation of taste buds is not enough for the onset of the taste transduction, which necessitates also mature taste cells. Moreover, the earlier maturation of palatal taste buds compared with the contiguous populations in the oral cavity evokes an evidence of their significant role in the transmission of gustatory information, especially in the early life of rat.

Large cholesterol granuloma arising from the frontal sinus--case report.

A 59-year-old male presented with a large cholesterol granuloma arising from the frontal sinus manifesting as a large, fluctuated, soft mass in his brow, compressing left eye. Skull radiography showed dilation of the frontal sinus. Computed tomography and magnetic resonance imaging revealed a cystic mass extending into the left orbit and anterior cranial fossa. Gross inspection at the frontal craniotomy showed mucinous, dark green fluid intermingled with shining material. The histological diagnosis was cholesterol granuloma with epithelial lining. Large cholesterol granuloma with facial deformity is always associated with bone and cosmetic problems. Wide opening of the frontal sinus followed by cyst wall removal and plastic repair of the skull is necessary.

Correlation of early CT signs in the deep middle cerebral artery territories with angiographically confirmed site of arterial occlusion.

BACKGROUND AND PURPOSE: Early CT signs in the deep middle cerebral artery (MCA) territories have been reported to be seen at the initial period of ischemia. We attempted to investigate the incidence of parenchymal hypodensity within 3 hours after ischemic onset among patients with angiographically proved embolic MCA occlusion and to assess the correlation of subtle hypodensity in the deep MCA territories with involvement of the lenticulostriate arteries in the presence of ischemia. METHODS: Fifty CT images obtained within 3 hours after onset of embolic MCA occlusion were retrospectively reviewed by three neurosurgeons who were aware of clinical features. Early CT signs in the deep MCA territories were divided into three grades according to their anatomic location: grade I, normal basal ganglia with hypodensity localized to the insula; grade II, partial obscuration of the posterolateral part of the putamen; and grade III, hypodensity of the entire lentiform nucleus. A grade I CT sign was considered to be a negative finding for lenticulostriate artery involvement, whereas grade II and III CT signs were considered to be positive findings. Site of occlusion and involvement of the lenticulostriate arteries were confirmed by angiography. RESULTS: Thirty-eight (76%) of 50 patients had early CT signs in the deep MCA territories. Sensitivity and specificity of a grade I CT sign indicating absence of lenticulostriate artery involvement in ischemia were 65% and 87%, respectively. On the other hand, sensitivity and specificity of grade II and grade III CT signs for presence of lenticulostriate artery involvement in ischemia were 77% and 100%, respectively. Grade II CT signs resulted from various sites of occlusion, whereas grade III was unequivocally predictive of proximal occlusion to all of the lenticulostriate arteries. CONCLUSION: Involvement of the lenticulostriate arteries may be presumed by precise evaluation of subtle, CT-revealed hypodensity in the deep MCA territories, even within 3 hours of ischemic onset.

Long-term enhancement of synaptic transmission induced by veratridine in rat CA3 hippocampal neurons.

Veratridine is a neurotoxin that induces persistent activation of sodium channels in excitable cells. We investigated the effects of this toxin on excitatory synaptic transmission in CA3 neurons of juvenile rat hippocampus using whole-cell patch-clamp and field-potential recordings. The population spikes evoked by electrical stimulation of the mossy fiber were gradually enhanced after washout of veratridine (0.3 microM), but they were not enhanced by the co-application of veratridine and an N-methyl-D-aspartate (NMDA) receptor antagonist (D-APV, 30 microM). When a pipette solution contained QX-314 that antagonized the effect of veratridine in the recorded neuron, oscillatory membrane depolarization appeared in the early stage during bath-application of veratridine and gradually decreased in the late stage. After washout of veratridine, however, the oscillatory depolarization was gradually restored and maintained for at least 3 h. This oscillatory depolarization was also abolished by D-APV. We suggest that the activation of NMDA receptors is involved in the veratridine-induced long-lasting enhancement in the excitatory synaptic transmission in rat CA3 hippocampal neurons.

The correlation between 99mTc-MIBI uptake and MIB-1 as a nuclear proliferation marker in glioma--a comparative study with 201Tl.

Technetium-99m methoxy-isobutylisonitrile (MIBI), like thallium-201 (201Tl), is a highly efficient agent for the diagnosis and monitoring of glioma tumors. Although 201Tl uptake is known to be partly associated with proliferative activity, little is known about the correlation between MIBI uptake and proliferation activity in gliomas. The current study was performed to assess the correlation between MIBI uptake and proliferative activities in gliomas, estimated by the monoclonal antibody to Ki-67 antigen (MIB-1) staining method. By comparing the results with those of 201Tl, we determined which tracer would be suitable for estimating proliferative activities. Twenty-four presurgical glioma patients (six with low-grade gliomas, five with anaplastic astrocytomas, and 13 with glioblastomas) were given MIBI and 201Tl SPECT. Early (10 min after injection) and delayed images (3 h after injection) were obtained for both MIBI and 201Tl scintigraphy. SPECT parameters, early ratio (ER), delayed ratio (DR), and retention index (RI) were obtained in both radiopharmaceuticals. All patients underwent subsequent surgical excision, and the specimens were immunostained for MIB-1. The proliferative activity was measured as a percentage positive nuclear area for MIB-1 (MI; MIB-1 index). To evaluate the relationship between the proliferative activity and SPECT parameters, we performed a correlation analysis. MI correlated with the MIBI uptake ratio (r = 0.75 for ER, and r = 0.7 for DR). Both DR and RI of 201Tl also correlated with MI, but weakly (r = 0.6 for DR, and. r = 0.59 for RI). There was no significant correlation between the MIB-1 index and the other parameters. MIBI-uptake parameters demonstrated a stronger positive correlation with the MIB-1 index than that of 201Tl. With the use of MIBI SPECT, we can estimate the proliferative activity of glioma noninvasively.

Differentiation of the lingual and palatal gustatory epithelium of the rat as revealed by immunohistochemistry of alpha-gustducin.

We used alpha-gustducin, a taste-cell-specific G protein to investigate the onset of taste transduction and its relation to the development of the palatal and lingual taste buds. Frozen cryostat and paraffin sections were prepared from the palatal and lingual gustatory epithelium of the rat from birth till postnatal day 21 (PN 21d). At PN 1-7d, alpha-gustducin-immunoreactive solitary ovoid or bipolar cells were scattered among the oral epithelium either horizontally along the oral surface or vertically oriented between the basal lamina and oral surface. In the circumvallate and foliate papillae, these cells became wrapped in alpha-gustducin-immunonegative cells surrounded by an extracellular space forming a bud-like structure. Simultaneously, different stages of typical taste buds were recognized, but alpha-gustducin was only expressed in some neonatally developed pored buds. At PN 1d, alpha-gustducin was expressed in pored taste buds with a relatively higher frequency recorded in the soft palate as compared with the nasoincisor, circumvallate, and foliate papillae. The immunoreactive cells were spindle shaped with elongated processes extending from the base to the pore of the taste buds. During the second week, the solitary cells could no longer be recognized while the total counts of immunoreactive cells within the taste buds gradually increased. We argue that taste transduction is essentially required from the time of birth and can be fulfilled by both of the solitary chemosensory cells, which are immunoreactive for alpha-gustducin and scattered in the oral epithelium, and the taste cells within the mature taste buds. Moreover, the onset of taste transduction accomplished by the palatal taste buds developed earlier than that achieved by taste buds in the circumvallate and foliate papillae.

Transient expression of heat shock protein (Hsp)25 in the dental pulp and enamel organ during odontogenesis in the rat incisor.

The expression of heat shock protein (Hsp) 25 during odontogenesis in the dental pulp and enamel organ of rat incisors was investigated by immunocytochemistry and confocal microscopy. In the process of dentin formation, immature odontoblasts first exhibited Hsp 25-immunoreactivity, and increased in immunointensity with the advance of their differentiation. In the dental pulp, in contrast, intense immunoreaction in the mesenchymal cells became weak or negative in parallel with the progress of cell differentiation. The immunoreaction for Hsp 25 in the enamel organ revealed a characteristic stage-related alteration during amelogenesis. In secretory ameloblasts, the immunoreaction for Hsp 25 was found throughout their cell bodies, intense reactivity being located near the proximal and distal terminal webs. At the maturation stage, ruffle-ended ameloblasts (RA) consistently showed Hsp 25-immunoreactivity throughout the cell bodies, whereas smooth-ended ameloblasts (SA) lacking a ruffled border were weak in immunoreaction at the distal cytoplasm. Other cellular elements of the enamel organ were negative. The subcellular localization of Hsp 25-immunoreactivity in this study appeared essentially identical to that of actin filaments as demonstrated by confocal microscopy using rhodamine-labeled phalloidin. These immunocytochemical data suggest that the Hsp 25 molecule is involved in reinforcement of the cell layer following cell movement during odontogenesis and in the formation and maintenance of the ruffled border of RA.