[Short-term exposure to gossypol causes reversible reproductive toxicity and nephrotoxicity in mice].

OBJECTIVE: To study the toxic effects of short-term exposure to gossypol on the testis and kidney in mice and whether these effects are reversible. METHODS: Twenty 7 to 8-week-old male mice were randomized into blank control group, solvent control group, gossypol treatment group and drug withdrawal group. In the former 3 groups, the mice were subjected to daily intragastric administration of 0.3 mL of purified water, 1% sodium carboxymethylcellulose solution, and 30 mg/mL gossypol solution for 14 days, respectively; In the drug withdrawal group, the mice were treated with gossypol solution in the same manner for 14 days followed by treatment with purified water for another 14 days. After the last administration, the mice were euthanized and tissue samples were collected. The testicular tissue was weighed and observed microscopically with HE and PAS staining; the kidney tissue was stained with HE and examined for mitochondrial ATPase activity. RESULTS: Compared with those in the control group, the mice with gossypol exposure showed reduced testicular seminiferous epithelial cells with rounded seminiferous tubules, enlarged space between the seminiferous tubules, interstitium atrophy of the testis, and incomplete differentiation of the spermatogonia. The gossypol-treated mice also presented with complete, non-elongated spermatids, a large number of cells in the state of round spermatids, and negativity for acrosome PAS reaction; diffuse renal mesangial cell hyperplasia, increased mesangial matrix, and adhesion of the mesangium to the wall of the renal capsule were observed, with significantly shrinkage or even absence of the lumens of the renal capsules and reduced kidney mitochondrial ATPase activity. Compared with the gossypol-treated mice, the mice in the drug withdrawal group showed obvious recovery of morphologies of the testis and the kidney, acrosome PAS reaction and mitochondrial ATPase activity. CONCLUSIONS: Shortterm treatment with gossypol can cause reproductive toxicity and nephrotoxicity in mice, but these toxic effects can be reversed after drug withdrawal.

[Updates in the pathological diagnosis of Pleural Malignant Mesothelioma in the WHO classification of thoracic tumors (5(th) edition)].

The WHO Classification of Thoracic Tumors (5(th) edition) mainly has the following changes in the chapter of pleural malignant mesothelioma. (1) The concept of mesothelioma in situ and its diagnostic method have been established for the first time; (2) The tumour grading of pleural malignant mesothelioma was added, it was divided into low grade and high grade according to the cellular atypia, mitotic activity and presence of necrosis. (3) The morphological features of pleural malignant mesothelioma was classified into architectural pattern, cellular and stromal features, the correlation between histological features and prognosis was refined, and some of the controversial cellular types have been reclassified. In this review, we introduced the changes of related pathologic diagnosis, in the WHO Classification of Thoracic Tumors (5(th) edition) and discussed its clinical significance.

The three-dimensional evaluation of positional change in mandibular condyle after intraoral vertical ramus osteotomy.

OBJECTIVES: The purpose of this study was to compare the changes of the condylar axis, the anteroposterior condylar position relative to the glenoid fossa, after intraoral vertical ramus osteotomy (IVRO). STUDY DESIGN: 21 patients diagnosed as skeletal class III malocclusion underwent IVRO and were followed according to the authors' postoperative management regimen. The three-dimensional positions of the condyles were evaluated by cone-beam computerized tomography (CBCT) at pre-op, post-op, and at follow-up. CBCT images were referenced to assess the condylar axis change and the anteroposterior condylar position in the glenoid fossa. A repeated-measures analysis of variance (P<0.05) also was performed. RESULTS: After surgery, both the axial condylar angles and the anteroposterior condylar position were significantly different (P<0.05). The coronal condylar axis rotated outwardly. The anteroposterior condylar position in the glenoid fossa had moved from the concentric to the anterior position. But the condyle changes between post-op and follow-up (P>0.05) were insignificant. CONCLUSIONS: With postoperative intermaxillary elastic traction, the condyles changed their positions physiologically for newly established jaw movement after IVRO.

[Root canal anatomy of maxillary second premolars at various ages observed by cone-beam CT].

Objective: To observe the morphological changes of root canals in maxillary second premolars at various ages by using cone-beam CT (CBCT) in order to provide imaging and theoretical reference for clinical treatments. Methods: The digital CBCT data of the maxillary second premolars in 440 cases from the patients in Department of Stomatology, The First Affiliated Hospital, Jinan University during March 2011 and December 2017 were collected. The CBCT images were divided into 4 groups according to the patients' ages: groups ≤20, 21-40, 41-60 and>60 years old, respectively. Changes of morphologies of root canals with aging including such parameters as types of the root canal, incidence of double root canals in single rooted teeth, distance between both root canal orifices of double rooted canals, and morphological change of the cross section of roots. Chi-square test and liner trend test were adopted in statistical analysis in the present study. Results: Most maxillary second premolars had only one root [95.2% (419/440)]. Type Ⅰ of the root canals was the most common type [57.3% (252/440)], and the following prevalent groups were type Ⅱ[16.8% (74/440)], type Ⅳ [10.2% (45/440)] and type Ⅲ [8.9% (39/440)]. The distribution of type Ⅰ~Ⅳ of the root canals were significantly different amongst various aged groups (P<0.05). Along with aging, the percentages of type Ⅰ decreased while type Ⅱ increased. However, there were no remarkable changes of type Ⅲ and type Ⅳ observed. The incidence of double canal in single rooted teeth gradually increased with aging especially in 20-year-old and above groups, e.g. 13.1% (13/99) in group of ≤20 years old and 45.0% (86/191) in group of 21-40 years old. However, there was no significant increase observed after the age of 40. The distance between two root canal orifices of double rooted canals became shorter with aging except in groups of 40-year-old and above. The morphologies of the cross sections of root canals in most groups were flat shaped [57.8% (1 121/1 938)] and oval shape [31.3% (607/1 938)]. Along with aging, the percentage of circular shape gradually increased while flat and oval shapes decreased. Conclusions: The morphology of root canal could be clearly showed by the CBCT images. Most maxillary second premolars had only one root and one apical foramen. Along with aging, the morphology of the root canals became more and more complicated.

Mutation of the rice TCM12 gene encoding 2,3-bisphosphoglycerate-independent phosphoglycerate mutase affects chlorophyll synthesis, photosynthesis and chloroplast development at seedling stage at low temperatures.

Glycolysis is a central metabolic pathway that provides energy and products of primary metabolites. 2,3-Biphosphoglycerate-independent phosphoglycerate mutase (iPGAM) is a key enzyme that catalyses the reversible interconversion of 3-phosphoglycerate (3-PGA) to 2-phosphoglycerate (2-PGA) in glycolysis. Low temperature is a common abiotic stress in rice production. However, the mechanism for rice iPGAM genes is not fully understood at low temperature. In this study, the rice mutant tcm12, with chlorosis, malformed chloroplasts and impaired photosynthesis, was grown at a low temperature (<20 °C) to the three-leaf stage, while the normal phenotype at 32 °C was used. Chlorophyll fluorescence analysis and transmission electron microscopy were used to examine features of the tcm12 mutant. The inheritance behaviour and function of TCM12 were then analysed thorough map-based cloning, transgenic complementation and subcellular localisation. The thermo-sensitive chlorosis phenotype was caused by a single nucleotide mutation (T→C) on the fifth exon of TCM12 (LOC_Os12g35040) encoding iPGAM, localised to both nucleus and membranes. In addition, TCM12 was constitutively expressed, and its disruption resulted in down-regulation of some genes associated with chlorophyll biosynthesis and photosynthesis at low temperatures (20 °C). This is the first report of the involvement of rice iPGAM gene in chlorophyll synthesis, photosynthesis and chloroplast development, providing new insights into the mechanisms underlying early growth of rice at low temperatures.

[Role of glycogen synthase kinase-3ß in development of severe hepatitis liver failure caused by hepatitis B virus].

OBJECTIVE: To investigate the role of glycogen synthase kinase-3ß (GSK3ß) in the development of severe hepatitis liver failure (SHLF) caused by the hepatitis B virus. METHODS: Twelve patients with chronic hepatitis B (CHB) (CHB group), 12 patients with SHLF caused by hepatitis B virus (SHLF group), and 8 normal subjects (control group), who were admitted to Beijing You'an Hospital from January 2009 to December 2011, were included in this study. Their liver tissues were collected to do some clinical examinations. The GSK3ß activity in the liver tissue was detected with a GSK3ß activity assay kit. Western blot was used to determine the expression of p-GSK3, total GSK3, and -actin. The paraffin sections of liver tissue were prepared for immunofluorescence assay. All data were expressed as mean±standard deviation, and comparison between groups was made by least significant difference t test. P < 0.05 was considered statistically significant. RESULTS: Western blot results showed that compared with the control group, the CHB group had a higher level of p-GSK3ß and the SHLF group had a significantly lower level of p-GSK3ß (P = 0.0342). The immunofluorescence assay results showed that the SHLF group had a significantly lower level of p-GSK3ß than the control group. GSK3ß activity assay results showed that compared with the control group, the CHB group had a significantly lower GSK3ß activity and the SHLF group had a significantly higher GSK3ß activity (P = 0.0289), which were consistent with the results of Western blot and immunofluorescence assay. CONCLUSION: GSK3 is activated in the development of SHLF, so it is an important signaling molecule in the pathogenesis of SHLF. Inhibiting its activity may play a role in the prevention and treatment of SHLF.

[Aging changes of the root canal morphology in maxillary first premolars observed by cone-beam computerized tomography].

OBJECTIVE: To observe the morphological changes of root canals with aging in maxillary first premolars by using cone-beam computerized tomography(CBCT)in order to facilitate endodontic management of root canals in various aged patients. METHODS: The digital CBCT data of the maxillary first premolars in 405 cases from the patients in Oral Medical Center of The First Affiliated Hospital, Jinan University from March 2011 to June 2015 were collected. The CBCT images were divided into 6 groups according to the patients' ages: groups 11-20, 21-30, 31-40, 41-50, 51-60 and >60 years-olds, respectively. Changes of morphologies of root canals with aging including such parameters as types of the root canal, incidence of double root canals in single rooted teeth, distance between both root canal orifices of double rooted canals, and morphological change of the cross section of roots. Chi-square test and liner trend test were adopted in statistical analysis in the present study. RESULTS: The distribution of various types of the root canals were significantly different amongst various aged groups(P<0.05). Type Ⅳ is the most common type(210/405, 51.8%), and the following groups were typeⅡ(65/405, 16.0%), typeⅠ(55/405, 13.6%)and type Ⅲ(27/405, 6.7%). Along with aging, the percentages of type Ⅰ and type Ⅲ decreased while type Ⅱ increased. However, there were no remarkable changes of type Ⅳ observed. The incidence of double rooted canals in single rooted teeth gradually increased with aging especially in 20-years-old and above groups, e.g. 51.7%(31/60)in group 11-20 years-olds and 83.0%(44/53)in group 21-30 years-olds. However, there was no significant increase observed after the age of 40. The distance between both root canal orifices of double rooted canals became shorter with aging except in groups of 40-years-olds and above. The morphologies of the cross sections in most aged groups were flat shaped(1 020/2 105, 48.5%)and oval shape(594/2 105, 28.2%). Along with aging, the percentage of circular shape gradually increased while flat and oval shapes decreased. CONCLUSIONS: The morphology of root canal could be clearly showed by the CBCT images. The change of morphologies of the root canals in maxillary first premolars was significantly related to aging. Along with aging, the morphology of the root canal became more and more complicated.

Three-dimensional architectural and structural analysis--a transition in concept and design from Delaire's cephalometric analysis.

The aim of this study was to present a systematic sequence for three-dimensional (3D) measurement and cephalometry, provide the norm data for computed tomography-based 3D architectural and structural cephalometric analysis, and validate the 3D data through comparison with Delaire's two-dimensional (2D) lateral cephalometric data for the same Korean adults. 2D and 3D cephalometric analyses were performed for 27 healthy subjects and the measurements of both analyses were then individually and comparatively analyzed. Essential diagnostic tools for 3D cephalometry with modified definitions of the points, planes, and measurements were set up based on a review of the conceptual differences between two and three dimensions. Some 2D and 3D analysis results were similar, though significant differences were found with regard to craniofacial angle (C1-F1), incisal axis angles, cranial base length (C2), and cranial height (C3). The discrepancy in C2 and C3 appeared to be directly related to the magnification of 2D cephalometric images. Considering measurement discrepancies between 2D and 3D Delaire's analyses due to differences in concept and design, 3D architectural and structural analysis needs to be conducted based on norms and a sound 3D basis for the sake of its accurate application and widespread adoption.

Visual pathway-related horizontal reference plane for three-dimensional craniofacial analysis.

OBJECTIVES: To construct three-dimensional (3D) horizontal reference planes based on visual pathway and to determine their stability and reliability by analyzing the structural patterns of normal and dysmorphology for 3D craniofacial analysis. SETTING AND SAMPLE POPULATION: Thirty-six subjects with maxillofacial dysmorphology and malocclusion, and eight normal controls. MATERIALS AND METHODS POPULATION: On the 3D computed tomographic images of the subjects, the visual pathway-based planes, including the orbital axis plane (OAP), visual axis plane (VAP), and the optical axis plane (OpAP), were constructed and evaluated. RESULTS: The OAP, but not the VAP and OpAP, showed the ideal relationship between the midsagittal and posterior maxillary plane, and properly described the different patterns of maxillofacial dysmorphology with craniofacial plane 1 of Delaire's analysis and the occlusal plane. CONCLUSIONS: The proposed visual pathway-related horizontal reference planes, and in particular the OAP, seem to correctly express the visual axis and the position of the head in natural head position and can be used as a horizontal reference plane for the 3D analysis of craniofacial dysmorphology and anthropology.

Multiple copies of 16s rRNA gene affect the restriction patterns and DGGE profile as revealed by analysis of genome database.

The use of 16S rRNA gene has been a "golden" method to determine the diversity of microbial communities in environmental samples, phylogenetic relationships of prokaryotes and taxonomic position of newly isolated organisms. However due to the presence of multiple heterogeneous 16S rRNA gene copies in many strains, the interpretation of microbial ecology via 16S rRNA sequences is complicated. Purpose of present paper is to demonstrate the extent to which the multiple heterogeneous 16S rRNA gene copies affect RFLP patterns and DGG E profiles by using the genome database. In present genome database, there are 782 bacterial strains in total whose genomes have been completely sequenced and annotated. Among the total strains, 639 strains (82%) possess multiple 16S rRNA gene copies, 415 strains (53%) whose multiple copies are heterogeneous in sequences as revealed by alignment, 236 strains (30%) whose multiple copies show different restrict patterns by CSP61 + Hinfl, MspI + Rsal or HhaI as analyzed in silico. Polymorphisms of the multiple copies in certain strains were further characterized by G + C% and phylogentic distances based on the sequences of V3 region, which are linked to DGGE patters. Polymorphisms of a few strains were shown as examples. Using artificial communities, it is demonstrated that the presence of multiple heterogeneous 16S rRNA gene copies potentially leads to over-estimation of the diversity of a community. It is suggested that care must be taken when interpreting 16S rRNA-based RFLP and DGGE data and profiling an environmental community.

Diffusion tensor imaging in patients with focal epilepsy due to cortical dysplasia in the temporo-occipital region: electro-clinico-pathological correlations.

PURPOSE: Diffusion tensor imaging (DTI) provides information about magnitude (diffusivity) and directionality (anisotropy, FA) of water diffusion. We explored the characteristics of pathology-proven cortical dysplasia (CD) in the posterior quadrant in a case series of three patients using DTI measures, to assess associated alterations in subcortical connectivity and correlate with in situ epileptogenicity, seizure propagation and functional outcome. METHODS: The FA maps were visually inspected by a Neuroradiologist blinded to clinical data and conventional MRI (PR) and tractography was performed to assess connectivity of the dysplastic cortex and correlate with seizure propagation on invasive EEG. RESULTS: Analysis of FA maps revealed reduced connectivity with reduced arborization and thinning of the fiber bundles between the subcortical WM and the dysplastic cortex in patients 1 and 2, confirmed on tractography. Fiber tracts reconstructed from regions underlying the ictal onset help explain ictal propagation patterns. In the two patients without preexisting visual field deficit, resections spared the optic radiation visible on the FA map. CONCLUSIONS: Diffusivity measures and visualization of tracts provide complementary information on white matter changes accompanying CD and may assist to explain ictal spread patterns. Careful correlation with measures of function will allow the assessment of the functional significance of various dysplastic lesions and may help to design resective strategies.

VDUP1 potentiates Ras-mediated angiogenesis via ROS production in endothelial cells.

Vitamin D3 up-regulated protein 1 (VDUP1) is a tumor suppressor of which expression is reduced in a variety of cancer cells, and enforced expression inhibits the tumor cell proliferation. It inhibits the activity of thioredoxin, thus contributing cellular ROS generation. Since ROS is a critical factor for angiogenesis, we investigated the role of VDUP1 in angiogenesis and endothelial proliferation. The expression of VDUP1 was upregulated by overexpression of an oncogene, Ras. Enforced expression of VDUP1 increases ROS production and proliferation of Ras-overexpressing endothelial cells. Overexpression of VDUP1 increases the resistance to the anchorage-dependent cell death and tube formation of the Ras-overexpressing endothelial cell. In addition, the removal of ROS by ROS scavenger attenuates the effect of VDUP1 on tube formation. These results suggest that VDUP1 is involved in Ras-mediated angiogenesis via ROS generation in endothelial cells.

Evidence for a basal temporal visual language center: cortical stimulation producing pure alexia.

BACKGROUND: Dejerine and Benson and Geschwind postulated disconnection of the dominant angular gyrus from both visual association cortices as the basis for pure alexia, emphasizing disruption of white matter tracts in the dominant temporooccipital region. Recently functional imaging studies provide evidence for direct participation of basal temporal and occipital cortices in the cognitive process of reading. The exact location and function of these areas remain a matter of debate. OBJECTIVE: To confirm the participation of the basal temporal region in reading. METHOD: Extraoperative electrical stimulation of the dominant hemisphere was performed in three subjects using subdural electrodes, as part of presurgical evaluation for refractory epilepsy. RESULTS: Pure alexia was reproduced during cortical stimulation of the dominant posterior fusiform and inferior temporal gyri in all three patients. Stimulation resulted in selective reading difficulty with intact auditory comprehension and writing. Reading difficulty involved sentences and words with intact letter by letter reading. Picture naming difficulties were also noted at some electrodes. This region is located posterior to and contiguous with the basal temporal language area (BTLA) where stimulation resulted in global language dysfunction in visual and auditory realms. The location corresponded with the visual word form area described on functional MRI. CONCLUSION: These observations support the existence of a visual language area in the dominant fusiform and occipitotemporal gyri, contiguous with basal temporal language area. A portion of visual language area was exclusively involved in lexical processing while the other part of this region processed both lexical and nonlexical symbols.

Hyper-methylation of RIZ1 tumor suppressor gene is involved in the early tumorigenesis of hepatocellular carcinoma.

The retinoblastoma protein-interacting zinc finger gene RIZ1 is a putative tumor suppressor gene, and the inactivation of the RIZ1 is frequently found in tumors through a loss of mRNA expression. In order to understand the role of RIZ1 inactivation in the tumorigenesis of hepatocellular carcinoma (HCC), we detected the RIZ1 promoter methylation status in 39 HCCs using a methylation specific PCR (MSP) method, and carried out LOH study with marker P704. We also assessed the associations between the methylation status and clinicopathological parameters, tumor size, tumor differentiation, and fractional allelic loss (FAL). The results showed that the RIZ1 promoter methylated both in advanced tumors (>3 cm), (18/31, 58.0%) and in early tumors (<3 cm), (4/8, 50.0%). There were 54.6% (12/22) tumors with hyper-methylation in the low FAL group and 45.5% (10/22) in the high FAL group. Moreover, the DNA methylation of the RIZ1 promoter was found not only in the poorly differentiated tumors (12/22, 54.6%), but also in the well differentiated tumors (10/22, 45.5%). Among the 22 HCCs (22/39, 56.4%) that showed hyper-methylation at the RIZ1 promoter region, 3 cases showed biallelic methylation. Interestingly, one case showed hyper-methylation on one allele and a loss of heterozygosity (LOH) on the other allele. In other words, 4 HCCs showed the biallelic inactivation of the RIZ1. These results suggest that the inactivation of the RIZ1 by DNA methylation at its promoter region is involved in the tumorigenesis of HCC, particularly in the early stage of disease.

Evaluation of clinical and serological data from Taenia solium cysticercosis patients in eastern Inner Mongolia Autonomous Region, China.

Neurocysticercosis (NCC) is one of the major causes of neurological disease in China. ELISA and immunoblotting using glycoproteins purified by preparative isoelectric focusing were used to detect human cysticercosis in Tongliao area, Inner Mongolia, China in 1998. Approximately 89% (39 of 44 inpatients and outpatients with suspected NCC at Tongliao City Hospital) were residents of Inner Mongolia. About 53% were male and 77% were of working age (18-59 years), and 32% were farmers. Immunoblotting and ELISA showed a high correlation. Of the 44 patients, 31 positive by cerebral computed tomography (CT) scan were confirmed serologically to have cysticercosis. In the ELISA, patients with no lesions by CT scan had lower OD values, similar to those of normal serum. These findings confirm that both ELISA and immunoblotting assays are sufficiently sensitive to detect asymptomatic or symptomatic cysticercosis patients.

SCAR and CAPS mapping of CRb, a gene conferring resistance to Plasmodiophora brassicae in Chinese cabbage ( Brassica rapa ssp. pekinensis).

Clubroot disease, caused by Plasmodiophora brassicae Wor., is highly damaging for Chinese cabbage. The CR (clubroot resistant) Shinki DH (doubled haploid) line of Chinese cabbage carries a single dominant gene, CRb, which confers resistance to the P. brassicae races 2, 4, and 8. An F(2) population derived from a cross between the CR Shinki DH line and a susceptible line, 94SK, was used to map the CRb gene. Inoculation of F(3) families with SSI (single-spore isolate) resulted in a 1:2:1 segregation ratio. Use of the AFLP technique combined with bulked segregant analysis allowed five co-dominant AFLP markers, and four and seven dominant AFLP markers linked in coupling and repulsion, respectively, to be identified. Six of the 16 AFLP markers showing low frequencies of recombination with the CRb locus among 138 F(2) lines were cloned. A reliable conversion procedure allowed five AFLP markers to be successfully converted into CAPS and SCAR markers. An F(2) population (143 plants) was analyzed with these markers and a previously identified SCAR marker, and a genetic map around CRb covering a total distance of 6.75 cM was constructed. One dominant marker, TCR09, was located 0.78 cM from CRb. The remaining markers (TCR05, TCR01, TCR10, TCR08, and TCR03) were located on the other side of CRb, and the nearest of these was TCR05, at a distance of 1.92 cM.

Activation of vanilloid receptor 1 (VR1) by eugenol.

The structural similarity of eugenol with capsaicin suggests that these two agents may share molecular mechanisms to produce their effects. We investigated the effects of eugenol in comparison with those of capsaicin using whole-cell patch clamp and Fura-2-based calcium-imaging techniques in a heterologous expression system and with sensory neurons. In vanilloid receptor 1 (VR1)-expressing human embryonic kidney (HEK) 293 cells and trigeminal ganglion (TG) neurons, eugenol activated inward currents, whereas capsazepine, a competitive VR antagonist, and ruthenium red (RR), a functional VR antagonist, completely blocked eugenol-induced inward currents. Moreover, eugenol caused elevation of [Ca(2+)](i), and this was completely abolished by both capsazepine and ruthenium red in VR1-expressing HEK 293 cells and TG neurons. Our results provide strong evidence that eugenol produces its effects, at least in part, via VR1 expressed by the sensory nerve endings in the teeth.

Hypermethylation in human cancers of the RIZ1 tumor suppressor gene, a member of a histone/protein methyltransferase superfamily.

The retinoblastoma protein-interacting zinc finger gene RIZ1 is a tumor suppressor gene and a member of a nuclear histone/protein methyltransferase superfamily. RIZ1 inactivation is commonly found in many types of human cancers and occurs through loss of mRNA expression, frameshift mutation, chromosomal deletion, and missense mutation. RIZ1 is also a tumor susceptibility gene in mice. We now show that loss of RIZ1 mRNA in human cancers is associated with DNA methylation of its promoter CpG island. Methylation of the RIZ1 promoter strongly correlated with lost or decreased RIZ1 mRNA expression in breast, liver, colon, and lung cancer cell lines as well as in liver cancer tissues. Treatment with the methylation inhibitor 5-aza-2'-deoxycytidine activated RIZ1 mRNA expression in cancer cells. Furthermore, methylation was found in 11 of 25 (44%) breast cancer specimens and 20 of 32 (62%) liver cancer specimens. Our results suggest that DNA methylation is a common mechanism in inactivating the RIZ1 tumor suppressor gene in human liver and breast cancers.

Participation of intracellular Ca(2+)/calmodulin and protein kinase(s) in the pathway of apoptosis induced by a Drosophila cell death gene, reaper.

To elucidate the apoptotic signaling pathway, we have generated a cell culture model: S2 cells stably transfected with a Drosophila cell death gene, reaper (rpr). Following rpr overexpression, caspase activation-mediated apoptotic cell death was induced in the cells. Apoptosis triggered by rpr required intracellular Ca(2+) ions and calmodulin. Furthermore, protein kinase inhibitors H-7 (a PKC, PKA, PKG, MLCK, and CKI inhibitor), calphostin C (a PKC inhibitor), or H-89 (a PKA and PKG inhibitor) completely blocked apoptosis induced by rpr, suggesting that some kind of serine/threonine protein kinase(s) act upstream of caspase in apoptotic pathway induced by rpr in S2 cells.