The limitations of gallium scintigraphy in the differentiation between benign and malignant salivary gland mass lesions.

The aim of this study was to evaluate the ability of gallium scintigraphy to differentiate between benign and malignant salivary gland mass lesions and to identify what types of lesions surpass its diagnostic utility. By considering the uptake of 67Ga, 193 salivary gland masses were graded visually as negative, weakly positive, moderately positive or strongly positive in comparison to the uptake in the nasal cavity and the liver. The uptake was compared with histopathological findings. Among 39 malignant tumours, uptake was positive in 31 (79%) (strongly positive in 18, moderately positive in seven and weakly positive in six) and uptake was negative in eight (21%). Adenoid cystic carcinoma was the most common malignant tumour in our study (11/39), and uptake was negative in five (45%) of these tumours. Malignant tumours did not differ significantly in size despite differences in uptake. Among 154 benign lesions, uptake was negative in 101 (66%) and positive in 53 (34%) (strongly positive in 12, moderately positive in 19 and weakly positive in 22). Out of 88 pleomorphic adenomas, 41 (47%) showed positive uptake. Sensitivity, specificity and accuracy for gallium study were 80%, 66% and 68%, respectively, when the malignancy criterion was weakly positive uptake. Accuracy was greatest (83%) when the criterion was strongly positive uptake, but this criterion failed to detect more than a half of malignant tumours (46% sensitivity). In conclusion, gallium scintigraphy had limitations in differentiating between benign and malignant salivary gland mass lesions. Adenoid cystic carcinomas and pleomorphic adenomas were the principal sources of false negative and false positive results, respectively.

Decreased expression of an ATP-binding cassette transporter, MRP2, in human livers with hepatitis C virus infection.

BACKGROUND/AIMS: To understand hepatic injury during the process of hepatitis viral infection, determination of liver-specific functions at molecular levels is critical. Because the transport of endogenous/exogenous toxic substances is an intrinsically important hepatic function, we examined whether expression of the ATP-binding cassette (ABC) transporter gene was affected in patients with hepatitis viral infection. METHODS: To determine which ABC transporter was expressed differently in patients with hepatic viral infection, we assayed the expression of MDR1, MDR3, MRP1, MRP2, and MRP3 in non-cancerous regions in the liver of 42 patients with hepatic tumors using both quantitative RT-PCR and immunological staining analysis, and compared the hepatic expression levels between patients with hepatitis viral infection and non-infected controls. RESULTS: Of the five ABC transporter genes studied, the mRNAs of MRP2 and MRP3 were highly expressed in the human liver. There was a significant reduction in MRP2 expression to 29% in the virus-infected liver. Treatment of hepatic cells with inflammatory cytokines resulted in decreased mRNA levels of MRP2 and decreased MRP2 promoter activity. CONCLUSIONS: The down-regulation of MRP2 might induce a failure in the transport of various genotoxic substances in the liver with hepatitis virus infection.

Enhanced expression of the human multidrug resistance protein 3 by bile salt in human enterocytes. A transcriptional control of a plausible bile acid transporter.

The enterohepatic circulation is essential for the maintenance of bile acids and cholesterol homeostasis. The ileal bile acid transporter on the apical membrane of enterocytes mediates the intestinal uptake of bile salts, but little is known about the bile salt secretion from the basolateral membrane of enterocytes into blood. In the basolateral membrane of enterocytes, an ATP-binding cassette transporter, multidrug resistance protein 3 (MRP3), is expressed, which has the ability to transport bile salts. We hypothesized that MRP3 might play a role in the enterohepatic circulation of bile salts by transporting them from enterocytes into circulating blood through the up-regulation of MRP3 expression, so we investigated the transcriptional control of MRP3 in response to bile salts. MRP3 mRNA levels were increased about 3-fold in human colon cells by chenodeoxycholic acid (CDCA), in a dose- and time-dependent manner. In the promoter assay, the promoter activity of MRP3 was increased about 3-fold over the basal promoter activity when treated with CDCA, and the putative bile salt-responsive elements exist in the region -229/-138 including two alpha-1 fetoprotein transcription factor (FTF)-like elements. Constructs with a specific mutation in the consensus sequence of FTF elements showed no increase in basal transcriptional activity following CDCA treatment. In electrophoretic mobility shift assay with nuclear extracts, specific binding of FTF to FTF-like elements was observed when treated with CDCA. The expression of FTF mRNA levels were also markedly enhanced in response to CDCA, and overexpression of FTF specifically activated the MRP3 promoter activity about 4-fold over the basal promoter activity. FTF thus might play a key role not only in the bile salt synthetic pathway in hepatocytes but also in the bile salt excretion pathway in enterocytes through the regulation of MRP3 expression. MRP3 may contribute as a plausible bile salt-exporting transporter to the enterohepatic circulation of bile salts.

Transfer of SV40 temperature-sensitive early gene into human epidermal keratinocytes by the recombinant adenovirus vector.

We constructed a recombinant adenovirus vector that contained the origin-defective SV40 early gene, coding temperature-sensitive T antigen. This vector transferred the SV40 early gene into human epidermal keratinocytes with high efficiency. T antigen conferred the ability of keratinocytes to grow with limited differentiation in the presence of serum and high calcium concentration at the permissive temperature (34 degrees C), although normal keratinocytes were induced to differentiate and stop growing under the same conditions. The serum/Ca++-resistant cells did not proliferate at the nonpermissive temperature (40 degrees C), indicating that they depended on T antigen for their proliferation. The temperature-sensitive T antigen dissociated from the tumor suppressor gene products, p53, at 40 degrees C. The serum/Ca++-resistant cells still had the ability to proceed to terminal differentiation when injected into SCID mice as cultured keratinocytes. However, they did not form an apparent basal layer. This indicated that the tissue remodeling process in the serum/Ca++-resistant keratinocytes was abnormal. All of these epidermoid cysts disappeared within 8 wk and no tumor developed for 6 mo. We consider that deltaE1/SVtsT is a useful tool to examine multistep carcinogenesis of human epithelial cells in vitro.

The human multidrug resistance protein 2 gene: functional characterization of the 5'-flanking region and expression in hepatic cells.

The human multidrug resistance protein 2 (MRP2), also termed as the canalicular multispecific organic anion transporter (cMOAT), is a member of the adenosine triphosphate-binding cassette transporter superfamily. In the liver, MRP2 mediates the multispecific efflux of various types of organic anions, including glucuronate, sulfate, and glutathione conjugates, across the canalicular hepatocyte membrane to the bile. To investigate how the MRP2 gene is expressed in liver cells, the 5'-flanking region of the human MRP2 gene was isolated from a human placental genomic library. Sequence analysis of the MRP2 promoter showed a number of consensus binding sites for both ubiquitous and liver-enriched transcription factors. Transfection of human hepatic HepG2 cells with a series of 5'-deleted promoter luciferase constructs identified a putative silencer element localized in the -1,659/-491 region and a liver-specific positive regulatory element localized in the -491/-258 region. This latter region contained the liver-abundant transcription factor CCAAT-enhancer binding protein beta (C/EBPbeta). The transcriptional activity of the promoter construct containing a mutation in the C/EBPbeta binding site was significantly decreased in HepG2 cells. This study suggests that C/EBPbeta (-356 to -343) may regulate the liver expression of the MRP2 gene.

The role of the labyrinth, proprioception and plantar mechanosensors in the maintenance of an upright posture.

The maintenance of an upright posture in man requires information from vision, the labyrinth, proprioception and plantar mechanosensors. In order to evaluate the role of the labyrinth, proprioception and plantar mechanosensors, stabilometry was performed in subjects with closed eyes. Ten patients with bilateral severe or complete labyrinthine paresis were studied, as well as 9 patients with severe proprioceptive disorders and 10 normal healthy persons whose plantar mechanosensors were anesthetized by hypothermia. Both the area of sway and the total locus length (accumulated shift distance length) were evaluated. On closing eyes, in patients with labyrinthine disorders demonstrated that the area of sway increased more than length. On the other hand, in patients with proprioceptive disorders, length increased more than the area. In plantar anesthetized subjects, similar to the labyrinthine disorder cases, the area of sway increased more than length. These findings suggest that the labyrinth is a main monitor of the area of body sway, while proprioception is a principle monitor of the velocity of body movement of sway (or locus length). The plantar mechanosensor monitors the area of body sway similar to the labyrinth, but works less than the labyrinth. The locus length is the distance per minute and reflects the velocity of body sway. Thus, the length per area is a parameter for the velocity of body sway per area. Since proprioceptive disorders increase both the locus length and the length per area, present findings suggest that if proprioception is damaged, the body begins to move faster. Compensated labyrinthine disorders have a tendency to increase the length per area, indicating that if a labyrinthine disorder is compensated, the body adapts and moves faster to maintain an upright posture.

Effects of the antihistaminergic drugs diphenhydramine and zolantidine on vestibular-induced hypothalamic neuronal activity in the guinea pig.

The mode of action of diphenhydramine in treating motion sickness is unknown. Using an electrophysiologic technique, we investigated the effects of intravenous diphenhydramine and zolantidine on the changes in neuronal activity produced by caloric stimulation in the hypothalamic paraventricular nucleus (PVN) in the guinea pig. Changes in neuronal activity were modulated by the administration of diphenhydramine in a high percentage of the neurons tested (71%), while zolantidine affected only a small number (29%). This finding reinforces the involvement of a histaminergic system in vestibular autonomic responses. The modulatory effect of diphenhydramine on PVN neuron activity may explain in part this drug's efficacy in treating motion sickness.

Huge hamartoma with inverted papilloma in the nasal cavity.

We report clinical experience in managing a 46-year-old Japanese man with long-standing nasal obstruction resulting from a huge left nasal mass. Computed tomography, magnetic resonance imaging and biopsy were used to make a provisional diagnosis of inverted papilloma. The mass was resected via a frontal approach combined with rhinotomy. Histopathologic examination of the resected specimen was consistent with a hamartoma that included an inverted papilloma on a portion of its surface. In addition to being rare tumors in the nasal cavity, we believe that our patient's tumor the largest nasal hamartoma ever reported.

Magnetic sensory cortical responses evoked by tactile stimulations of the human face, oral cavity and flap reconstructions of the tongue.

Magnetoencephalography (MEG) is a procedure that analyzes the magnetic responses of neurons. An MEG system with a 37-channel superconductivity quantum interference device (SQUID) was used to record magnetic signals from the human brain in response to tactile stimulations of the face and oral cavity. Six normal individuals were studied as well as three patients who had undergone hemiglossectomies as treatment for carcinoma of the tongue and reconstruction with a pectoralis major myocutaneous flap. When the locations of the magnetic responses having latencies of 40 ms from onset of tactile stimulation were identified, these corresponded to the primary somatosensory cortex. In patients whose tongues had been reconstructed with a pectoralis major myocutaneous flap, the magnetic response upon stimulation of the flap was recorded in a sensory cortical area identical to that corresponding to the tongue. MEG systems such as the one described permit functional mapping of the cerebral cortex on stimulating the face and oral cavity.

Overexpression of p53 nuclear protein in premalignant and malignant laryngeal lesions.

To investigate the role of p53 nuclear protein mutations in the initiation and progression of laryngeal carcinoma, 111 premalignant and malignant laryngeal lesions (19 specimens with hyperkeratosis and 92 with carcinoma) were studied immunohistochemically. Over-expression of p53 was observed in 8 cases (42%) of laryngeal hyperkeratosis and 44 cases (47%) of laryngeal carcinoma. However, the expression of p53 showed no relationship to patients' clinical courses. Our study confirms that p53 overexpression can be found in laryngeal carcinogenesis and is an early event but not a useful prognostic marker.

Comparison of survival rates of patients with nasopharyngeal carcinoma treated with radiotherapy, 5-fluorouracil and vitamin A ("FAR" therapy) vs FAR therapy plus adjunctive cisplatin and peplomycin chemotherapy.

The overall survival rate (OSR) of 36 patients with nasopharyngeal carcinomas (NPC) treated at Kyushu University hospital between 1983 to 1992 was analyzed. As primary treatment, 16 patients received a combination therapy of 5-fluorouracil, vitamin A, and radiation (FAR therapy); two patients received radiotherapy only; 18 patients received FAR therapy plus adjunctive systemic chemotherapy consisting of cisplatin and peplomycin. The radiation dose to the nasopharynx was 6000 to 7050 cGy while that to the neck was 4000-6000 cGy. The 5-year OSR of all the patients was 49%. Histological type (moderately differentiated squamous cell carcinoma) and patient age (> or = 55) were found to be significant prognostic factors for a worse OSR. Although survival decreased with increasing T stage, no significant difference was observed. The 5-year OSR of the patients treated with FAR therapy was 53% and was 51% with FAR therapy plus chemotherapy. Compared to FAR therapy alone, adjunctive chemotherapy did not increase OSR of the patients with NPC.

Induction of apoptosis in maxillary sinus cancer cells by 5-fluorouracil, vitamin A and radiation (FAR) therapy.

The triple combination of 5-fluorouracil (5-FU), vitamin A and radiation (FAR therapy) has been used since 1972 to treat malignant tumors of the head and neck at Kyushu University. Using nick end labeling of tumor specimens, cells of human maxillary sinus carcinomas were observed previously to undergo apoptosis in response to FAR therapy. The present study evaluated the in vitro effects of FAR therapy on a human maxillary sinus cancer (IMC-4) cell line. We further compared the effects of FAR therapy on this cell line with those effects seen on tissue samples taken from patients with maxillary sinus cancers. DNA electrophoresis and electron microscopic examination of the IMC-4 cells after treatment with FAR therapy revealed typical apoptotic features. The effects of 50-100 micrograms/ml 5-FU, 10(-4) M all-trans-retinoic acid and radiation to 6 Gy on IMC-4 cells were evaluated by trypan blue dye exclusion and a cell colony formation assay. 5-FU and radiation caused direct cell death, while vitamin A mainly inhibited cell growth. The combination of these treatment as FAR therapy synergistically enhanced cell death and inhibited cell growth. Flow cytometry demonstrated that FAR-treated cells were arrested in the G1 phase of the cell cycle before undergoing apoptosis. To further investigate possible biological parameters influencing a tumor's apoptotic sensitivity, we also examined the expression of p53 in human maxillary sinus cancer cells and analyzed the relationship between p53 expression and apoptosis. However, no relationship was found between these two markers at the time point studied.

[A case of adenoid cystic carcinoma in epipharynx which responded to radiation and chemotherapy].

A 45-year-old man visited our hospital complaining of a right hearing loss. The head and neck examination was unremarkable except for right secretory otitis media. Fiberoptic examination of the nose and the epipharynx revealed a mass. Transnasal biopsy was performed and histologic analysis revealed adenoid cystic carcinoma. Invasion of the sphenoid sinus was identified by MRI. He was treated with FAR therapy (5-FU, Vit A, Radiation) and chemotherapy (intra-arterial infusion of ADM). Severe side effects were not seen during treatment. After treatment, the majority of the tumor mass disappeared on fiberoptic examination and MRI. On repeat biopsy specimens, no cancer cells were found. The patient was alive with no evidence of disease as of August 1997.

Neuronal responses to vestibular stimulation in the guinea pig hypothalamic paraventricular nucleus.

We investigated the effects of caloric stimulation on neuronal activity in the hypothalamic paraventricular nucleus (PVN) in anesthetized guinea pigs. Hot water stimulation of the contralateral labyrinth produced excitation in 29.4% of the PVN neurons tested, while cold water produced excitation in 22.2% of the neurons. Hot water resulted in inhibition of 22.4% of the neurons and cold water inhibition of 24.7% of the neurons. Intracranial vestibular nerve section greatly reduced responsiveness of the PVN neurons to caloric stimulation, indicating that the majority of the responses observed were vestibular in origin. The response pattern of the individual PVN neurons was similar following hot and cold water stimulation and after stimulation of the contralateral and ipsilateral labyrinths. These results suggest that the PVN neurons receive vestibular afferents bilaterally according to the intensity of vestibular stimulation, with the information received probably integrated in the hypothalamus to participate in vestibulo-autonomic reflexes.

Effects of stimulation of the vestibular nuclei on posterior hypothalamic neuron activity in guinea pigs.

To clarify the differences among the four main vestibular nuclei in the vestibulo-autonomic reflex, we examined the effects of electrical stimulation of superior, lateral, medial and descending vestibular nuclei (SVN, LVN, MVN and DVN) on posterior hypothalamic area (PHA) neurons in the guinea pig. Ipsi- and contralateral SVN stimulation produced excitation in 30% and 25% of the PHA neurons tested, respectively. Twenty percent of the PHA neurons showed an excitatory response to ipsilateral LVN stimulation while 60% of the neurons tested responded to contralateral LVN stimulation, including excitation of 36% and inhibition of 24%. MVN and DVN stimulation produced little change in PHA neuron activity. These findings suggest that vestibular information processed in the SVN and the LVN is conveyed to the hypothalamus and may then contribute to activation of the vestibulo-autonomic reflex.

Convergence of olfactory and nasotrigeminal inputs and possible trigeminal contributions to olfactory responses in the rat thalamus.

To elucidate the role of trigeminal input on the olfactory system, field-evoked potentials were measured following electrical stimulation of the nasociliary branch of the trigeminal nerve in the olfactory-related structures in the rat brain. Significant potential changes were recorded in the mediodorsal nucleus of the thalamus and the lateral hypothalamic area. In the mediodorsal nucleus of the thalamus, the neurons responding to olfactory bulb electrical stimulation also responded to trigeminal nerve stimulation. Single neuronal responses of mediodorsal thalamic neurons following odorant stimulation were enhanced by blockade of the trigeminal nerve with procaine. These results suggest that olfactory and trigeminal pathways converge on the same neural elements within the mediodorsal nucleus of the thalamus and that the trigeminal input may modulate olfactory input in this nucleus.

Effects of locus ceruleus and olfactory bulb stimulation on rat olfactory tubercle neuron activity.

The effects of electrical stimulation of the olfactory bulb and the locus ceruleus on olfactory tubercle neurons were examined in rat models. Ipsilateral stimulation of the olfactory bulb produced excitation in 31% of olfactory tubercle neurons tested and inhibition in 17%. Twenty-two percent of the olfactory tubercle neurons were excited, whereas 9% were inhibited by ipsilateral stimulation of the locus ceruleus. Contralateral stimulation of the locus ceruleus produced similar responses in the same neuron entities. A negative-positive evoked potential was recorded in the olfactory tubercle after ipsilateral and contralateral stimulation of the locus ceruleus. Thirty-three percent of the olfactory tubercle neurons that responded orthodromically or antidromically to stimulation of the olfactory bulb were excited by ipsilateral stimulation of the locus ceruleus. In contrast, only 10% responded with excitation to ipsilateral stimulation of the locus ceruleus among the olfactory tubercle neurons that were unresponsive to stimulation of the olfactory bulb. These findings suggest that olfactory tubercle neurons that receive input from or sending output to the olfactory bulb are influenced by the noradrenergic system of the locus ceruleus. A possible role of the olfactory tubercle in olfactory transduction will also be discussed.

Projections from the ventral tegmental area to the olfactory tubercle in the rat.

The projection between the ventral tegmental area (VTA) and the olfactory tubercle (OT) was examined electrophysiologically in the rat. Stimulation of the olfactory bulb (OB) determined if the OT neurons were olfactory-related. Ipsilateral VTA stimulation produced a change in neuronal activity in 77% of the neurons tested, with 41% being inhibited, 24% excited, and 12% had mixed response. Contralateral VTA stimulation produced changes in only 38%. Intravenous administration of haloperidol was used in examination of the role of dopamine in this neural connection. The results suggest that the VTA-induced inhibitory response on OT neurons is mediated by dopamine, whereas excitatory responses are not. The VTA inhibitory influence projects primarily to olfactory-related neurons, since 60% of olfactory-related OT neurons were inhibited--as compared to 34% of non-olfactory-related neurons. This study documents electrophysiologically the VTA-OT connection and suggests that the dopaminergic input may modulate olfactory information projected to the OT from the OB. It also supports the concept that the OT acts as an integration center in central olfactory processing.

Glucagon-related peptides in the rat hypothalamus.

Immunohistochemically, nerve fibers and terminals reacting with anti-N-terminal-specific but not with anti-C-terminal-specific glucagon antiserum were observed in the following rat hypothalamic regions: paraventricular nucleus, supraoptic nucleus, anterior hypothalamus, arcuate nucleus, ventromedial hypothalamic nucleus and median eminence. Few fibers and terminals were demonstrated in the lateral hypothalamic area and dorsomedial hypothalamic nucleus. Radioimmunoassay data indicated that the concentration of gut glucagon-like immunoreactivity was higher in the ventromedial nucleus than in the lateral hypothalamic area. In food-deprived conditions, this concentration increased in both these parts. This was also verified in immunostained preparations in which a marked enhancement of gut glucagon-like immunoreactivity-containing fibers and terminals was observed in many hypothalamic regions. Several immunoreactive cell bodies were found in the ventromedial and arcuate nuclei of starved rats. Both biochemical and morphological data suggest that glucagon-related peptides may act as neurotransmitters or neuromodulators in the hypothalamus and may be involved in the central regulatory mechanism related to feeding behavior and energy metabolism.