The Characteristic Changes in Hepatitis B Virus X Region for Hepatocellular Carcinoma: A Comprehensive Analysis Based on Global Data

Objectives

Mutations in hepatitis B virus (HBV) X region (HBx) play important roles in hepatocarcinogenesis while the results remain controversial. We sought to clarify potential hepatocellular carcinoma (HCC)-characteristic mutations in HBx from HBV genotype C-infected patients and the distribution of those mutations in different disease phases and genotypes.

Methods

HBx sequences downloaded from an online global HBV database were screened and then classified into Non-HCC or HCC group by diagnosis information. Patients'' data of patient age, gender, country or area, and viral genotype were also extracted. Logistic regression was performed to evaluate the effects of mutations on HCC risk.

Results

1) Full length HBx sequences (HCC: 161; Non-HCC: 954) originated from 1115 human sera across 29 countries/areas were extracted from the downloaded 5956 HBx sequences. Genotype C occupied 40.6% of Non-HCC (387/954) and 89.4% of HCC (144/161). 2) Sixteen nucleotide positions showed significantly different distributions between genotype C HCC and Non-HCC groups. 3) Logistic regression showed that mutations A1383C (OR: 2.32, 95% CI: 1.34-4.01), R1479C/T (OR: 1.96, 95% CI: 1.05-3.64; OR: 5.15, 95% CI: 2.53-10.48), C1485T (OR: 2.40, 95% CI: 1.41-4.08), C1631T (OR: 4.09, 95% CI: 1.41-11.85), C1653T (OR: 2.58, 95% CI: 1.59-4.19), G1719T (OR: 2.11, 95% CI: 1.19-3.73), and T1800C (OR: 23.59, 95% CI: 2.25-247.65) were independent risk factors for genotype C HBV-related HCC, presenting different trends among individual disease phases. 4) Several genotype C HCC risk mutations pre-existed, even as major types, in early disease phases with other genotypes.

Conclusions

Mutations associated with HCC risk were mainly located in HBx transactivation domain, viral promoter, protein/miRNA binding sites, and the area for immune epitopes. Furthermore, the signatures of these mutations were unique to disease phases leading to HCC, suggesting molecular counteractions between the virus and host during hepatocarcinogenesis.     

Chimpanzees' choices of prey between two sympatric species ofMacrotermes in the Campo Animal Reserve,Cameroon

Details are presented relating to chimpanzees' choices between two sympatric species of termites,Macrotermes lillijeborgi andM. vitrialatus, as food in the Campo Animal Reserve, southwest Cameroon, West Africa. An attempt was made to determine the various factors that affected such choices. The two species of termites seemed to have almost the same value in terms of ecological factors. However, chimpanzees fed almost exclusivelyonM. lillijeborgi, using digging sticks and fisching probes, during the study period which extended from the end of August to the middle of January, with their feeding activity showing peak at the beginning of the rainy season. By contrast,M. vitrialatus was rarely eaten in spite of the ease with which such prey could be obtained, namely, by desctruction of termite mounds by hand, without the need for tools. The reason that the chimpanzees discriminated between the two species of termite cannot be explained in terms of ecological factors such as size of prey, seasonal differences in termite activity, etc. Sticks used as tools were fairly uniform in size and character, mainly because of physical constraints related to the structure of termite mounds, and the brush-like ends of sticks seemed to be incidentaly byproducts of the chimpanzee's choice of plant species. Ecological factors could provide chimpanzees with a basis for the use of some kind of tool and help them modify it, while other factors, for example, something akin to leisure or the chimpanzee's interest in use of a tool, could provide an opportunity for inventing some tool-using behavior or for maintaining such behavior. These different factors, not being exclusive of one another, might affect the invention and maintenance of tool using-behavior at different phases. It is possible that chimpanzee's choice of prey may not always be the most efficient or appropriate in a given ecological situation.     

Developmental changes in mother-offspring grooming in Japanese macaques

A cross-sectional analysis was made of developmental changes in mother-offspring grooming in Japanese macaques, Macaca,. fuscata. When offspring were immature, time spent grooming by their mothers decreased with offspring age. Soliciting by offspring increased steadily with age, in contrast to their successful soliciting, which decreased gradually until early adolescence. This is in accord with the hypothesis that grooming is one form of post-weaning maternal investment, which may entail behavioral conflict between mothers and immature offspring. On the other hand, mothers spent much more time for grooming of their adult female offspring than for their adolescent male and female offspring. It is argued that grooming by mothers may shift from a form of maternal investment in their offspring to a benefit to be exchanged reciprocally with them. © 1995 Wiley-Liss, Inc.     

Fatal attack on an infant by an adult female tonkean macaque

We describe a fatal attack on a 10-month-old female infant by an adult female Tonkean macaque (Macaca tonkeana)in a semi-free-ranging group. During an intense conflict involving many group members, an adult female, the second most dominant in the group, attacked the infant of a lower-ranking female. The attacker remained near the victim for approximately 20 min, grabbing and biting it repeatedly, until the mother retrieved the infant. She died 2 days later. An autopsy revealed no lethal injury;the main cause of death was starvation or dehydration or both. We discuss social conditions under which a fatal attack by an adult female might occur.     

Norepinephrine triggers Ca2+-dependent exocytosis of 5-hydroxytryptamine from rat pinealocytes in culture

5-hydroxytryptamine (5-HT) is a precursor and a putative modulator for melatonin synthesis in mammalian pinealocytes. 5-HT is present in organelles distinct from l-glutamate-containing synaptic-like microvesicles as well as in the cytoplasm of pinealocytes, and is secreted upon stimulation by norepinephrine (NE) to enhance serotonin N-acetyltransferase activity via the 5-HT2 receptor. However, the mechanism underlying the secretion of 5-HT from pinealocytes is unknown. In this study, we show that NE-evoked release of 5-HT is largely dependent on Ca2+ in rat pinealocytes in culture. Omission of Ca2+ from the medium and incubation of pineal cells with EGTA-tetraacetoxymethyl-ester inhibited by 59 and 97% the NE-evoked 5-HT release, respectively. Phenylephrine also triggered the Ca2+-dependent release of 5-HT, which was blocked by phentolamine, an alpha antagonist, but not by propranolol, a beta antagonist. Botulinum neurotoxin type E cleaved 25 kDa synaptosomal-associated protein and inhibited by 50% of the NE-evoked 5-HT release. Bafilomycin A1, an inhibitor of vacuolar H+-ATPase, and reserpine and tetrabenazine, inhibitors of vesicular monoamine transporter, all decreased the storage of vesicular 5-HT followed by inhibition of the NE-evoked 5-HT release. Agents that trigger L-glutamte exocytosis such as acetylcholine did not trigger any Ca2+-dependent 5-HT release. Vice versa neither NE nor phenylephrine caused synaptic-like microvesicle-mediated l-glutamate exocytosis. These results indicated that upon stimulation of a adrenoceptors pinealocytes secrete 5-HT through a Ca2+-dependent exocytotic mechanism, which is distinct from the exocytosis of synaptic-like microvesicles.     

Wnt proteins promote neuronal differentiation in neural stem cell culture

Wnt signaling is implicated in the control of cell growth and differentiation during CNS development from studies of mouse and chick models, but its action at the cellular level has been poorly understand. In this study, we examine the in vitro function of Wnt signaling in embryonic neural stem cells, dissociated from neurospheres derived from E11.5 mouse telencephalon. Conditioned media containing active Wnt-3a proteins are added to the neural stem cells and its effect on regeneration of neurospheres and differentiation into neuronal and glial cells was examined. Wnt-3a proteins inhibit regeneration of neurospheres, but promote differentiation into MAP2-positive neuronal cells. Wnt-3a proteins also increase the number of GFAP-positive astrocytes but suppress the number of oligodendroglial lineage cells expressing PDGFR or O4. These results indicate that Wnt-3a signaling can inhibit the maintenance of neural stem cells, but rather promote the differentiation of neural stem cells into several cell lineages.     

A nonradioisotope, enzymatic microplate assay for in vivo evaluation of 2-deoxyglucose uptake in muscle tissue

To determine 2-deoxy-D-glucose (2DG) and 2-deoxy-D-glucose 6-phosphate (DG6P) in mouse tissue after injection of 2DG, we have developed a novel assay. This assay is a simple procedure involving incubation of samples with four independent, single reaction mixtures followed by measurement of fluorescence. From differences between the values obtained with the four reactions, each of glucose, glucose 6-phosphate, 2DG and DG6P were able to be quantified in a sensitive manner. Using this assay system, glucose and 2DG in blood and DG6P-accumulation in muscle were easily determined. Therefore, this assay may be useful for measuring in vivo glucose uptake without the use of radioisotopes.