cAMP-GEFII is a direct target of cAMP in regulated exocytosis.

Although cAMP is well known to regulate exocytosis in many secretory cells, its direct target in the exocytotic machinery is not known. Here we show that cAMP-GEFII, a cAMP sensor, binds to Rim (Rab3-interacting molecule, Rab3 being a small G protein) and to a new isoform, Rim2, both of which are putative regulators of fusion of vesicles to the plasma membrane. We also show that cAMP-GEFII, through its interaction with Rim2, mediates cAMP-induced, Ca2+-dependent secretion that is not blocked by an inhibitor of cAMP-dependent protein kinase (PKA). Accordingly, cAMP-GEFII is a direct target of cAMP in regulated exocytosis and is responsible for cAMP-dependent, PKA-independent exocytosis.

Adjustment to cancer and the information needs of people with lung cancer.

OBJECTIVE: Although typically high, the need for information varies between cancer patients. Few studies, however, have examined the factors that predict patient information needs. This study investigated the influence of different styles of adjustment to cancer on information needs. It was proposed that adjustment styles can be defined in terms of goal pursuit and that adjustment influences information needs as these also arise from goal pursuit. METHOD: Seventy-three lung cancer patients were recruited at their first appointment with their radiation oncologist. Participants completed the Patient Information Needs Questionnaire measuring Disease Orientated (DO) information and Action Orientated (AO) information, the Mini-Mental Adjustment to Cancer Scale, and a purpose-built measure of cancer-related personal goals. RESULTS: High levels of the adjustment styles, Fighting Spirit and Anxious Preoccupation, were related to a high need for DO information (p=0.042 and 0.023, respectively). Conversely, high levels of the adjustment style Cognitive Avoidance was related to a low need for DO information (p=0.041). High levels of Anxious Preoccupation were also positively related to a high need for AO information (p=0.018). Support for the proposed theoretical model was also found: information goals predicted information needs and mediated the relationship between Fighting Spirit and DO information need. CONCLUSIONS: These findings suggest that information needs vary as a function of adjustment to cancer. Consequently information provision to cancer patients could be more appropriately tailored by attending to how a patient is adjusting to their diagnosis of cancer.

ATP-sensitive K+ channels in the hypothalamus are essential for the maintenance of glucose homeostasis.

Glucose-responsive (GR) neurons in the hypothalamus are thought to be critical in glucose homeostasis, but it is not known how they function in this context. Kir6.2 is the pore-forming subunit of K(ATP) channels in many cell types, including pancreatic beta-cells and heart. Here we show the complete absence of both functional ATP-sensitive K+ (K(ATP)) channels and glucose responsiveness in the neurons of the ventromedial hypothalamus (VMH) in Kir6.2-/- mice. Although pancreatic alpha-cells were functional in Kir6.2-/-, the mice exhibited a severe defect in glucagon secretion in response to systemic hypoglycemia. In addition, they showed a complete loss of glucagon secretion, together with reduced food intake in response to neuroglycopenia. Thus, our results demonstrate that KATP channels are important in glucose sensing in VMH GR neurons, and are essential for the maintenance of glucose homeostasis.

Isolation and characterization of the Candida albicans gene for mRNA 5'-triphosphatase: association of mRNA 5'-triphosphatase and mRNA 5'-guanylyltransferase activities is essential for the function of mRNA 5'-capping enzyme in vivo.

The amino acid sequence of the Saccharomyces cerevisiae mRNA 5'-triphosphatase (TPase) diverges from those of higher eukaryotes. In order to confirm the sequence divergence of TPases in lower and higher eukaryotes, the Candida albicans gene for TPase was identified and characterized. This gene designated CaCET1 (C. albicans mRNA 5'-capping enzyme triphosphatase 1) has an open reading frame of 1.5 kb, which can encode a 59-kDa protein. Although the N-terminal one-fifth of S. cerevisiae TPase (ScCet1p) is missing in CaCet1p, CaCet1p shares significant sequence similarity with ScCet1p over the entire region of the protein; the recombinant CaCet1p, which was expressed as a fusion protein with glutathione S-transferase (GST), displayed TPase activity in vitro. CaCET1 rescued CET1-deficient S. cerevisiae cells when expressed under the control of the ADH1 promoter, whereas the human capping enzyme derivatives that are active for TPase activity but defective in mRNA 5'-guanylyltransferase (GTase) activity did not. Yeast two-hybrid analysis revealed that C. albicans Cet1p can bind to the S. cerevisiae GTase in addition to its own partner, the C. albicans GTase. In contrast, neither the full-length human capping enzyme nor its TPase domain interacted with the yeast GTase. These results indicate that the failure of the human TPase activity to complement an S. cerevisiae cet1delta null mutation is attributable, at least in part, to the inability of the human capping enzyme to associate with the yeast GTase, and that the physical association of GTase and TPase is essential for the function of the capping enzyme in vivo.

Group impressions as dynamic configurations: the tensor product model of group impression formation and change.

Group impressions are dynamic configurations. The tensor product model (TPM), a connectionist model of memory and learning, is used to describe the process of group impression formation and change, emphasizing the structured and contextualized nature of group impressions and the dynamic evolution of group impressions over time. TPM is first shown to be consistent with algebraic models of social judgment (the weighted averaging model; N. Anderson, 1981) and exemplar-based social category learning (the context model; E. R. Smith & M. A. Zárate, 1992), providing a theoretical reduction of the algebraic models to the present connectionist framework. TPM is then shown to describe a common process that underlies both formation and change of group impressions despite the often-made assumption that they constitute different psychological processes. In particular, various time-dependent properties of both group impression formation (e.g., time variability, response dependency, and order effects in impression judgments) and change (e.g., stereotype change and group accentuation) are explained, demonstrating a hidden unity beneath the diverse array of empirical findings. Implications of the model for conceptualizing stereotype formation and change are discussed.

Regional differences in extracellular choline dependency of acetylcholine synthesis in the rat brain.

Acetylcholine (ACh) synthesis under conditions of restricted extracellular choline uptake was investigated in order to clarify the procurement of choline for ACh synthesis using slices of several regions of the rat brain. Extracellular choline-independent ACh synthesis was observed in the hippocampus, frontal cortex and caudate putamen, which contain cholinergic nerve terminals, whereas little or no synthesis was observed in the medial septum or basal nucleus of Meynert, which contain cholinergic cell bodies. These results indicate that cholinergic nerve terminals, but not the cell bodies, may be able to synthesize choline for ACh biosynthesis. Extracellular choline-dependent ACh synthesis was observed in all regions examined. In the presence of 10 microM choline, the highest content of newly synthesized ACh and the proportionate increase in ACh compared with the unreleasable fraction (basal level) were observed in the caudate putamen. In the frontal cortex, although the level of synthesized ACh was low, the proportionate increase in ACh was high. In contrast, in the medial septum and basal nucleus of Meynert, high levels of ACh with a low proportionate increase compared with basal levels were observed. In the presence of hemicholinium-3, extracellular choline was also taken up for ACh synthesis in all regions examined, the level being especially high in the frontal cortex and medial septum. The present results indicate that the manner of choline procurement for ACh synthesis is heterogeneous among the various regions of the rat brain.

Hemicholinium-3-resistant choline uptake system linked to acetylcholine synthesis in the rat hippocampus.

Extracellular choline-dependency of acetylcholine (ACh) synthesis was examined in rat hippocampal slices. In the presence of hemicholinium-3 (HC-3, 5 microM), extracellular choline-dependent ACh synthesis appeared to consist of two different components. The first component (saturated at up to 50 microM choline) was dependent on the HC-3-resistant choline uptake system having a low capacity for choline in comparison with the high-affinity choline uptake system (HACU). The second component, observed in the presence of more than 50 microM choline, was considered due to competitive inhibition of HACU by HC-3. HACU-dependent ACh synthesis seemed to be saturated in the presence of up to 2 microM choline. These results indicate that both the HACU and HC-3-resistant choline uptake systems are linked to ACh synthesis in the hippocampus.

[3H]2-(4-phenylpiperidino)cyclohexanol (AH5183) binding to synaptosomes and subcellular fractions obtained from rat brain.

We investigated the binding of [3H]AH5183 (2-(4-phenylpiperidino)cyclohexanol) to rat brain synaptosomes and subcellular fractions. A high content of specific binding was observed in crude synaptosomes obtained from the striatum, cerebral cortex and hippocampus. The highest density of subcellular binding sites was observed in the synaptic vesicle-rich fraction. The affinities of AH5183 binding to crude synaptosomes and the synaptic vesicle-rich fraction were almost equivalent, but the density of binding sites was higher in the synaptic vesicle fraction. The present findings indicate that [3H]AH5183 binding to both synaptosomes and the synaptic vesicle-rich fraction is useful as a cholinergic marker, and that for quantitative studies, binding to the latter fraction is more preferable.

Acetylcholine synthesis is modulated by acetylcholine content of cytosolic fraction but not by that of releasable fraction.

Synthesis and release of acetylcholine (ACh) in the rat hippocampal slices were examined to clarify the mechanism of modulation of ACh synthesis. Treatment with 2-(4-phenylpiperidino)cyclohexanol (AH5183, 50 microM), an inhibitor of ACh transport from cytosol to synaptic vesicles, inhibited the increase in ACh content of the membrane-bound fraction which is readily releasable, but did not affect the cytosolic ACh content. Under these conditions, the total ACh content reached a plateau value. These results indicate that ACh synthesis is modulated by cytosolic ACh content but not by the vesicular fraction.

Culture, gender, and self: a perspective from individualism-collectivism research.

Individualism and collectivism are often equated with independent vs. interdependent, agentic vs. communal, and separate vs. relational self-construals. Although these same concepts have been used to characterize both cultural and gender differences, a perspective of cultural evolution suggests it is unlikely. A division of labor within society may produce gender differences, but this cannot explain cultural differences. A study of self-construal involving 5 cultures (Australia, the United States, Hawaii, Japan, and Korea) shows that differences between these cultures are captured mostly by the extent to which people see themselves as acting as independent agents, whereas gender differences are best summarized by the extent to which people regard themselves as emotionally related to others.

An immunohistochemical study of TNF-alpha in optic nerves from AIDS patients.

PURPOSE: Both in vitro and in vivo studies have implicated a role for tumor necrosis factor (TNF-alpha) in the pathology of demyelinating diseases. The purpose of this study was to address the hypothesis that TNF-alpha is a mediator of AIDS-related optic nerve injury and to determine the cell types involved in the proliferation of TNF-alpha in the AIDS optic nerve. METHODS: Ten optic nerves from seven patients with AIDS, and three from persons who were HIV negative were stained, using the indirect immunoperoxidase method. Six of the ten AIDS optic nerves were positive for cytomegalovirus (CMV), but the remainder did not have abnormal fundus findings. RESULTS: In all the optic nerves from AIDS patients with or without CMV retinitis, the vast majority of astrocytes stained strongly for TNF-alpha. Microglial cells (MPS-derived macrophages) varied from not staining to staining strongly positive for TNF-alpha. However, oligodendrocytes were not labeled positively for TNF-alpha. Some endothelial cells also stained for TNF-alpha. Examination of normal optic nerves and controls did not reveal any cell type that stained positively for TNF-alpha. CONCLUSIONS: The present study supports the contention that TNF-alpha is a major mediator of AIDS-associated optic neuropathy. HIV infection induces the production of TNF-alpha in macrophages and astrocytes, which probably causes demyelination and other neuronal damage.

Role of intracellular esterases in the production of esters by Acetobacter pasteurianus.

Esters are the major flavor compounds produced by Acetobacter sp. during vinegar production. The two genes encoding the esterases in the bacteria were disrupted, and the effects of the disruptions studied. When cultured in the presence of ethanol, the est1 gene-disrupted mutant (DE1K) did not produce any ethyl acetate or isoamyl acetate. However, the disruption of est2 did not affect the ester production. Ethyl acetate production by N-23 (pME122P) and DE1K (pME122P), which contain est1, was 1.7-fold higher than that by the wild type, N-23. On analyzing the relationship between ethyl acetate production and the extracellular ethanol and acetic acid concentrations, we found that the highest amount of ethyl acetate was produced when the molar ratio of ethanol and acetic acid was 1:1. These results indicate that the ester production by Acetobacter sp. is mostly catalyzed by the intracellular esterase, esterase-1, with ethanol and acetic acid used as the substrates.

Cloning and characterization of ethanol-regulated esterase genes in Acetobacter pasteurianus.

The esterase encoding genes, est1 and est2, were cloned from Acetobacter pasteurianus. Nucleotide sequence analysis of est1 revealed a gene of 954 bp, and est1 coded for an arylesterase with a molecular weight of 34863 Da consisting of 317 amino acids. The est2 gene contained an open reading frame composed of 1221 bp encoding an esterase with a molecular weight of 43389 Da consisting of 406 amino acids. The est1 gene showed some similarity, but the est2 gene showed no significant homology to other esterases reported in various microorganisms. Northern blot analysis of total RNA from A. pasteurianus revealed that transcription of the est1 gene was induced only when the cells were grown in a medium containing ethanol, and suggested that the est1 transcript is monocistronic. In contrast, transcription of the est2 gene was repressed in the presence of ethanol. In the absence of ethanol, expression of the est2-mRNA, capable of encoding a multiple number of proteins, was revealed by Northern blot analysis. In addition, deletion analysis indicated that the 5'-region of the est2 gene contained a cis-acting domain for est2 transcriptional regulation. Analysis of the est1 promoter using the chloramphenicol acetyltransferase gene as a reporter gene showed that the promoter within the 305-bp fragment upstream of the ATG initiation codon was responsible for the transcription in cells grown in the presence of ethanol. Primer extension analysis of est1-mRNA showed that the transcription initiation site was 49 bp upstream from the ATG initiation codon. The results of a gel mobility shift assay indicated that there is a regulatory protein related to est1 regulation, which may have some relation to the ethanol resistance of Acetobacter sp.

A successful treatment for hepatocellular carcinoma with atrial tumor thrombus.

A 66-year-old man with an advanced hepatocellular carcinoma and tumor thrombus extending into the right atrium was treated by transcatheter arterial infusion of lipiodol and aclarubicin. This brought about a remarkable reduction of the tumor and the disappearance of the right atrial tumor thrombus. The tumor was then radically resected by hepatic posterior segmentectomy with combined resection of the right hepatic vein, where the tumor thrombus remained. He is doing well without any signs of recurrence 22 months after the operation.

The Ishikawa classification of cavernous sinus lesions by clinico-anatomical findings.

PURPOSE: The Jefferson classification has been used to localize cavernous sinus lesions. However, this classification occasionally showed dissociation between identified localization and clinical findings. We investigated the clinical applicability of the newly proposed Ishikawa classification based on serial topographic sections of human cavernous sinus and the clinical findings. METHODS: In the Ishikawa classification, the cavernous sinus is divided into three portions, that is, anterior, middle, and posterior, demarcated by the location of the intracranial orifice of the optic canal and the entry of the maxillary nerve into the cavernous sinus. A total of 162 patients with cavernous sinus lesions were classified using both the Jefferson and the Ishikawa classifications and the clinical applicability of these two classifications was studied. Characteristics of the localization of lesions were also examined in each etiological type. RESULTS: By the Jefferson classification, 11% of the 162 patients had the anterior type of lesion, 12% the middle, 8% the posterior type, and 69% the unclassifiable type. However, by the Ishikawa classification, 35% had the anterior type, 10% the middle type, 22% the posterior type, 5% the whole type, and 28% the unclassifiable type of lesion. Furthermore, the Ishikawa classification revealed that the etiology of the anterior type was mainly inflammation, and that the etiology of the posterior and whole types was tumors. CONCLUSION: The Ishikawa classification is clinically useful to identify and classify the localization of cavernous sinus lesions.

The theory of reasoned action and cooperative behaviour: it takes two to use a condom.

The applicability of the Theories of Reasoned Action and Planned Behaviour to the cooperative behaviour of condom use were examined. Seventy-one male and 78 female students, all sexually active unmarried heterosexuals aged 17 to 21 years, gave information about their intentions for the next sexual encounter, as well as their attitude, subjective norm, expectancy-value attitude and subjective norm (including normative beliefs for their sexual partner), and their past behaviour with respect to condom use. After their next sexual encounter, they completed a questionnaire on their actual condom use. Results indicated that when behavioural conditions including the availability of a condom and an agreement with the partner to use it were satisfied, intention interacted with past behaviour to predict actual behaviour. These results imply that intentions which are consistent with past behaviour are stable enough to be carried out in the face of the interpersonal dynamics of a sexual encounter. Further, normative belief for the sexual partner had a direct influence on attitudes, subjective norm and intention. Neither the Theory of Reasoned Action nor the Theory of Planned Behaviour can fully explain these results, which point to the need for further theoretical inquiry into the dynamics of cooperative behaviour.

[Mercury concentration of fish in Tokyo Bay and the surrounding sea area].

Total mercury in the muscles of three fish species was analyzed in fish caught in Tokyo Bay and the surrounding sea areas, Sagami Bay and Choshi. Tokyo Bay is a semi-closed sea area surrounded by Tokyo, Kanagawa and Chiba prefectures. Sagami Bay and Choshi are open to the Pacific Ocean. A total of 412 fish consisting of northern whiting (Sillago japonica), flatfish (Limanda yokohamae) and sardine (Sardinops melanosticta) were caught in these areas over a 6 months period from November 1998 to April 1999. Total mercury concentration ranged from 0.008-0.092 microgram/g (wet wt.) in northern whiting, 0.006-0.065 microgram/g in flatfish and 0.001-0.045 microgram/g in sardine. All concentrations were below the restriction limit of fish mercury in Japan, 0.4 microgram/g of total mercury concentration. A significant correlation was found between mercury concentrations and body length or body weight in northern whiting and flatfish, irrespective of the sea area. A correlation was also found between mercury concentration in fish and their feeding habits: among the 3 species caught in the same area, crustacean feeding northern whiting had the highest, polychaete feeding flatfish moderate, and plankton feeding sardine had the lowest mercury concentration. In a comparison of mercury concentration in the same species caught in different sea areas, a higher concentration was noted in fish caught in the semi-closed sea area of Tokyo Bay, than in fish caught in the open sea areas of Sagami Bay and Choshi. This difference was most marked in fish caught at the bottom of Tokyo Bay and we considered that the mercury concentration of seawater and sediment in these areas was the cause of mercury accumulation in fish. These findings suggest that improved water quality control and environmental monitoring is necessary in semi-closed sea areas such as Tokyo Bay.

[Horizontal gaze palsy due to lesion of the abducens nucleus detected with magnetic resonance imaging].

A case of unilateral horizontal gaze palsy is reported. The lesion of the abducens nucleus was clearly detected with magnetic resonance imaging. In the early stage, the adduction of the right eye and abduction of the left eye were completely limited and the vestibulo-ocular reflex was also impaired. However, during the recovery stage, the improvement of limitation of abduction was delayed compared with that of adduction. We suggest that this phenomenon is useful as a differential diagnosis between lesions of the abducens nucleus and of the paramedian pontine reticular formation.

[A pediatric case of ophthalmoplegic migraine with recurrent oculomotor nerve palsy].

PURPOSE: A young patient of ophthalmoplegic migraine with recurrent oculomotor nerve palsy is reported. CASE: A 4-year-old girl came to our hospital complaining of recurrent left ptosis and double vision which completely resolved within a few days. She had a history of drowsiness after vomiting every two months. Left oculomotor nerve palsy with internal ophthalmoplegia was observed. Electroencephalography revealed mild changes, but computed tomography, single photon emission computed tomography, and cerebrovascular Doppler-ultrasound examination demonstrated no abnormalities. This oculomotor nerve palsy completely improved on the next day. RESULTS: Thereafter, left episodic mydriasis and oculomotor nerve palsy occurred twice each. However, the frequency and symptoms of migraine were remarkably reduced by topical administration of 0.25% timolol maleate twice daily to both eyes. CONCLUSION: In isolated oculomotor nerve palsy in childhood, ophthalmoplegic migraine should be also considered. We emphasize that a detailed case history is very important to its diagnosis.

[Ishikawa's classification of cavernous sinus lesions by clinico-anatomical findings].

Jefferson's classification (1938) has been used to localize lesions of the cavernous sinus. However, we think this classification is sometimes incorrect in identifying localization based on clinico-anatomical evidence. We investigated the efficacy of the newly proposed Ishikawa's classification. Based on the analysis on serial sections of human cavernous sinus, it classifies the locus of the lesion into three groups: anterior, middle, and posterior, corresponding to the location of the intracranial orifice of the optic canal and of the penetration of the maxillary nerve into the cavernous sinus. The subjects were 162 cases with cavernous sinus lesions. They were classified by the two methods. There was a total of 111 cases (69%) of unclassified cases in Jefferson's classification and 45 (28%) in Ishikawa's classification. The anterior type was frequently encountered in cases caused by inflammations and both posterior and whole types were caused by tumors in Ishikawa's classification. It was concluded that Ishikawa's classification is clinically useful to identify and classify the localization of cavernous sinus lesions.