Comprehensive analysis of genetic alterations and their prognostic impacts in adult acute myeloid leukemia patients.

To clarify the cooperative roles of recurrently identified mutations and to establish a more precise risk classification system in acute myeloid leukemia (AML), we comprehensively analyzed mutations in 51 genes, as well as cytogenetics and 11 chimeric transcripts, in 197 adult patients with de novo AML who were registered in the Japan Adult Leukemia Study Group AML201 study. We identified a total of 505 mutations in 44 genes, while only five genes, FLT3, NPM1, CEBPA, DNMT3A and KIT, were mutated in more than 10% of the patients. Although several cooperative and exclusive mutation patterns were observed, the accumulated mutation number was higher in cytogenetically normal AML and lower in AML with RUNX1-RUNX1T1 and CBFB-MYH11, indicating a strong potential of these translocations for the initiation of AML. Furthermore, we evaluated the prognostic impacts of each sole mutation and the combinations of mutations and/or cytogenetics, and demonstrated that AML patients could be clearly stratified into five risk groups for overall survival by including the mutation status of DNMT3A, MLL-PTD and TP53 genes in the risk classification system of the European LeukemiaNet. These results indicate that the prognosis of AML could be stratified by the major mutation status in combination with cytogenetics.

High expression of N-acetylglucosaminyltransferase IVa promotes invasion of choriocarcinoma.

BACKGROUND: Gestational trophoblastic diseases (GTDs) are related to trophoblasts, and human chorionic gonadotropin (hCG) is secreted by GTDs as well as normal placentas. However, the asparagine-linked sugar chains on hCG contain abnormal biantennary structures in invasive mole and choriocarcinoma, but not normal pregnancy or hydatidiform mole. N-acetylglucosaminyltransferase-IV (GnT-IV) catalyses ß1,4-N-acetylglucosamine branching on asparagine-linked oligosaccharides, which are consistent with the abnormal sugar chain structures on hCG. METHODS: We investigated GnT-IVa expression in GTDs and placentas by immunohistochemistry, western blot, and RT-PCR. We assessed the effects of GnT-IVa knockdown in choriocarcinoma cells in vitro and in vivo. RESULTS: The GnT-IVa was highly expressed in trophoblasts of invasive mole and choriocarcinoma, and moderately in extravillous trophoblasts during the first trimester, but not in hydatidiform mole or other normal trophoblasts. The GnT-IVa knockdown in choriocarcinoma cells significantly reduced migration and invasive capacities, and suppressed cellular adhesion to extracellular matrix proteins. The extent of ß1,4-N-acetylglucosamine branching on ß1 integrin was greatly reduced by GnT-IVa knockdown, although the expression of ß1 integrin was not changed. In vivo studies further demonstrated that GnT-IVa knockdown suppressed tumour engraftment and growth. CONCLUSION: These findings suggest that GnT-IVa is involved in regulating invasion of choriocarcinoma through modifications of the oligosaccharide chains of ß1 integrin.

A case report of placental mesenchymal dysplasia with an increased VEGF-D expression.

The pathogenesis of placental mesenchymal dysplasia (PMD) remains unclear. This report presents a case of PMD with a female fetus complicated with intrauterine growth restriction (IUGR). The ultrasound findings were similar to molar pregnancies, but PMD was suspected based on the presence of low ß-hCG levels and a normal karyotype. After delivery, pathological examination of the placenta showed dilated villi and thick-walled vessels lacking trophoblast proliferation, which thus led to a diagnosis of PMD. The VEGF-D (Xp22.31) mRNA expression was found to have increased in the abnormal villi. Whether this is an incidental or X-linked gene specific event in, IUGR complicated, PMD pathogenesis warrants further investigation of VEGF-D expression in PMD.

Heterogeneity of macrophage populations and expression of galectin-3 in cutaneous wound healing in rats.

The aim of this study was to investigate the properties of macrophages that infiltrated the sites of cutaneous wound healing in rats between 1 and 26 days post wounding (dpw). During the inflammation phase (1-3 dpw), ED1(+) (CD68(+)) macrophages with enhanced lysosomal activity dominated. From 5 to 7 dpw there was formation of granulation tissue as indicated by the presence of myofibroblasts expressing α-smooth muscle actin. At this stage, ED2(+) (CD163(+)) macrophages, capable of producing inflammatory factors, were dominant. The majority of ED1(+) macrophages expressed galectin-3, a regulator of fibrosis. Corresponding to the increased numbers of ED1(+) and ED2(+) macrophages at 3-9 dpw, there was increased expression of genes encoding transforming growth factor-ß1 (a major fibrogenic factor), monocyte chemoattractant protein-1 and colony stimulating factor-1. These macrophage-related factors might contribute to inflammation and formation of granulation tissue. OX6(+) macrophages expressing class II molecules of the major histocompatibility complex became predominant in the healing stages (15-26 dpw), indicating important roles for antigen-presenting cells in tissue remodelling. The OX6(+) macrophages were most likely derived from ED1(+) macrophages. The results of this study show that infiltration of phenotypically- and functionally-distinct macrophage populations characterizes different stages of the wound healing process.

The effects of changing position and angle of the proximal segment after intraoral vertical ramus osteotomy.

The authors evaluated changes in position and angle of the proximal segment, including the condyle, after intraoral vertical ramus osteotomy (IVRO) with and without a Le Fort I osteotomy to verify whether displacement of the proximal segment could induce postoperative complications. Changes in condylar angle, ramus angle, and displacement of proximal segment were measured pre- and postoperatively. The position of the temporomandibular joint (TMJ) disc was examined pre- and postoperatively. Trigeminal nerve hypoesthesia in the lower lip was assessed bilaterally. The postoperative horizontal condylar angle was significantly smaller than the preoperative one on the deviated and non-deviated sides (P<0.0001). The postoperative coronal condylar angle was significantly larger than the preoperative one on the deviated side (P=0.0483). The postoperative sagittal ramus angle was larger than the preoperative one on the deviated (P<0.0001) and non-deviated (P=0.00005) side. Most joints with an anteriorly-displaced disc with and without reduction improved on the non-deviated side; 5 of 16 joints improved on the deviated side. Results suggest the position and angle of the proximal segment, including the condyle, could change after IVRO. This could be associated with symptomatic improvement in TMJ, and extreme medial displacement of the proximal segment could delay recovery from lower lip hypoesthesia.

The effect of mandibular setback or two-jaws surgery on pharyngeal airway among different genders.

Cephalometric studies show significant gender differences in the size of the pharyngeal airway space. This study aimed to investigate and compare morphologic changes after mandibular setback or two-jaws surgery on the pharyngeal airway in men and women using computed tomography (CT). The sample included 34 women and 13 men diagnosed with Class III skeletal deformities, who had been treated by mandibular setback or bimaxillary surgery (maxillary advancement and mandibular setback). Anteroposterior, lateral and cross-sectional area dimensions of the airway, at the level of soft palate and base of tongue, were measured pre- and postoperatively on CT images. In the mandibular setback group, the anteroposterior and cross-sectional area of the pharyngeal airway at the level of the soft palate and base of tongue were significantly reduced for men or women (P<.05). In the two-jaws surgery group, only midsagittal anteroposterior dimensions at the same levels were significantly decreased for men or women (P<.05). The difference between any values measured between men and women who received bilateral sagittal split ramus osteotomy setback surgery or two-jaws surgery for the treatment of class III anteroposterior discrepancy were not statistically significant (P>.05). This study suggests that oropharyngeal airway measurements, important for airway patency, do not demonstrate sex dimorphism.

Effectiveness of preoperative concurrent chemoradiation therapy (CCRT) for locally advanced adenocarcinoma of cervix.

AIM: To determine the efficacy of preoperative concurrent chemoradiation therapy (CCRT) to improve the prognosis of locally advanced adenocarcinoma of the uterine cervix. METHODS: Twenty-five patients with clinical stage IB2-IVB adenocarcinoma of the cervix were received preoperative CCRT. The CCRT protocol included: external radiotherapy to the pelvis: 39.6 Gy; intra-arterial or intravenous infusion of 70 mg/m2 cisplatin, days 1 and 22; 24-h continuous intravenous infusion of 700 mg/m2 5-FU, days 1-4 and 22-25. Two weeks after the end of CCRT, patients underwent restaging followed by appropriate surgery with pelvic lymphadenectomy. RESULTS: The overall clinical response rate was 96% (24/25), with a complete response (CR) in 12/25 patients and partial response (PR) in 12/25. On pathological examination, 5 of 19 patients (26%) undergoing surgery showed a pathological CR, 13 patients showed a PR, and 1 patient no change (NC) in their disease. Grade 3 or 4 hematological toxicity was observed in 15 patients. Grade 3 gastrointestinal toxicity was observed in 8 patients. The median follow-up period was 34 months (range, 6-69). The 5-year overall survival (OS) rate was 84%, and the progression-free survival (PFS) rate was 76%. CONCLUSIONS: Preoperative CCRT improves the survival of patients with locally advanced adenocarcinoma of the cervix, with manageable toxicities.

Twist expression in patients with cervical cancer is associated with poor disease outcome.

BACKGROUND: Twist, a basic helix--loop-helix transcription factor, has been reported to be associated with the development and progression of human cancer. We examined the distribution and expression of Twist in cervical cancer to examine its clinical significance. PATIENTS AND METHODS: We examined the distribution and expression of Twist in 101 cervical cancer specimens and determined the association between their expression and the clinico-pathological features observed, including patient outcome. RESULTS: Of the 101 specimens, 55 cases were negative for Twist immuno-expression, whereas 46 were positive. When categorized into negative versus positive expression, Twist was not associated with any of the clinico-pathological parameters examined. Positive Twist expression significantly predicted poorer overall survival (OS) and progression-free survival (PFS) when compared with negative expression (P < 0.01). In the multivariate analyses, positive Twist expression was an independent prognostic factor for OS (P < 0.05). CONCLUSIONS: Our data imply that positive Twist expression seems to be a useful marker in patients with cervical cancer likely to have an unfavorable clinical outcome.

Expression of N-acetylglucosaminyltransferase V in endometrial cancer correlates with poor prognosis.

N-acetylglucosaminyltransferase V (GnT-V) is an enzyme that catalyses beta1-6 branching of N-acetylglucosamine on asparagine-linked oligosaccharides of cell proteins. The present study aimed to investigate GnT-V expression and its prognostic significance in endometrial cancer. N-acetylglucosaminyltransferase V expression was studied by immunohistochemistry in 74 surgically resected endometrial cancers, and the staining intensity was evaluated. High GnT-V expression in tumour cells was found in 43 (58.1%) of the 74 cases, and was positively correlated with advanced patient age, histological grade, and lymph vascular space involvement. Patients with high GnT-V expression had significantly impaired overall survival and progression-free survival (PFS) (P=0.0041 and P=0.0023, respectively) compared to patients with low expression of GnT-V. On multivariate analysis, GnT-V expression was an independent prognostic factor for PFS (P=0.0364). beta1-6 branching of asparagine-linked oligosaccharides was also detected in GnT-V-positive endometrial cancer cells by leukoagglutinating phytohaemagglutinin (L(4)-PHA) staining, and the molecular size of the major glycoproteins recognised by L(4)-PHA was approximately 60-200 kDa by lectin blot analysis. These results suggested that high GnT-V expression was correlated with an unfavourable clinical outcome, and that GnT-V is involved in the malignant potential of endometrial cancer by increasing the synthesis of beta1-6 branching of asparagine-linked oligosaccharides.

Use of the Sonopet ultrasonic curettage device in intraoral vertical ramus osteotomy.

This study was designed to evaluate the usefulness of the Sonopet UST-2001 (Miwatec Co., Ltd., Kawasaki, Kanagawa, Japan) ultrasonic curettage device, and to assess the outcome after intraoral vertical ramus osteotomy (IVRO). Thirteen Japanese adults (age range 20-41 years, mean age 29.6 years) presented with jaw deformities diagnosed as mandibular prognathism and asymmetry; they all underwent IVRO of the mandible. This procedure was followed by ultrasonic bone curettage using the Sonopet to make a guiding notch or groove in the lateral cortex of the ramus without damaging the vessels and nerves. After surgery, the osteotomy line was evaluated by three-dimensional computed tomography. In all patients, osteotomy with the Sonopet device was achieved safely, with minimal bleeding and no major complications. The distal segment could be moved into its ideal position and all patients achieved their ideal profiles. Ultrasonic bone curettage is a safe method for making a guiding groove, without damage to surrounding tissue, prior to complete IVRO.

A pure nongestational choriocarcinoma of the ovary diagnosed with short tandem repeat analysis: case report and review of the literature.

Nongestational pure choriocarcinoma of the ovary is a very rare germ cell tumor. Except in women before menarche, determination of the origin is very difficult without genetic analysis. We present a pure nongestational choriocarcinoma arising in the left ovary of a 19-year-old woman. Following surgery, pathologic findings of the tumor demonstrated pure choriocarcinoma without combination of other germ cell tumor elements. We confirmed its nongestational origin by DNA polymorphism analysis at 15 short tandem repeat loci. Multiple courses of chemotherapy with methotrexate, etoposide, and actinomycin-D were effective for this case. DNA polymorphism analysis is useful to determine genetic origin in pure choriocarcinoma of the ovary.

Changes in occlusal force after mandibular ramus osteotomy with and without Le Fort I osteotomy.

The purpose of this study was to evaluate the differences in bite force changes and occlusal contacts after sagittal split ramus osteotomy (SSRO) and intraoral vertical ramus osteotomy (IVRO) with and without Le Fort I osteotomy. Sixty female patients with diagnosed mandibular prognathism with or without asymmetry were divided into four groups (SSRO, IVRO, SSRO with Le Fort I osteotomy and IVRO with Le Fort I osteotomy). Bite force and occlusal contacts were measured preoperatively and at 1, 3, 6 and 12 months after surgery with pressure-sensitive sheets. The differences among surgical procedures were examined statistically. Maximum bite force and occlusal contacts returned to preoperative levels after between 3 and 6 months. Regarding time-dependent changes in bite force and occlusal contact area, there were no significant differences among the groups. In conclusion, this study suggests that the combination of IVRO or SSRO and Le Fort I osteotomy does not affect postoperative time-dependent changes.

Condylar and disc positions after intraoral vertical ramus osteotomy with and without a Le Fort I osteotomy.

The purpose of this study was to examine the changes in temporomandibular joint (TMJ) morphology and clinical symptoms after intraoral vertical ramus osteotomy (IVRO) with and without a Le Fort I osteotomy. Of 50 Japanese patients with mandibular prognathism with mandibular and bimaxillary asymmetry, 25 underwent IVRO and 25 underwent IVRO in combination with a Le Fort I osteotomy. The TMJ symptoms and joint morphology, including disc tissue, were assessed preoperatively and postoperatively by magnetic resonance imaging and axial cephalogram. Improvement was seen in just 50% of joints with anterior disc displacement (ADD) that received IVRO and 52% of those that received IVRO with Le Fort I osteotomy. Fewer or no TMJ symptoms were reported postoperatively in 97% of the joints that received IVRO and 90% that received IVRO with Le Fort I osteotomy. Postoperatively, there were significant condylar position changes and horizontal changes in the condylar long axis on both sides in the two groups. There were no significant differences between improved ADD and unimproved ADD in condylar position change and the angle of the condylar long axis, although distinctive postoperative condylar sag was seen. These results suggest that IVRO with or without Le Fort I osteotomy can improve ADD and TMJ symptoms along with condylar position and angle, but it is difficult to predict the amount of improvement in ADD.

Recurrent multinodular neurilemmoma of the female upper lip.

Neurilemmoma is the most commonly encountered nerve sheath tumour of the oral cavity. It generally appears as a single encapsulated nodule. The tongue is involved most frequently and the lip rarely. The tumour is usually uninodular. Multinodular neurilemmoma of the upper lip is very rare and has been reported in only one patient. This is the first reported case of multinodular neurilemmoma in the upper lip of a female.

A comparison of postoperative hypoesthesia between two types of sagittal split ramus osteotomy and intraoral vertical ramus osteotomy, using the trigeminal somatosensory-evoked potential method.

The purpose of this study was to evaluate hypoesthesia of the lower lip using trigeminal somatosensory-evoked potential following 2 types of sagittal split ramus osteotomy (SSRO) and intraoral vertical ramus osteotomy (IVRO). There were 30 patients with mandibular prognathism, with and without asymmetry, who were divided into three groups: the Obwegeser method (Ob) group, the Obwegeser-Dal Pont method (ODP) group and the intraoral vertical ramus osteotomy (IVRO) group. The trigeminal somatosensory-evoked potential was recorded in the region of the lower lip and evaluated preoperatively and postoperatively. The average recovery periods from lower lip hypoesthesia in the IVRO and the Ob group were significantly shorter than in the ODP group (P<0.05). In conclusion, IVRO showed the earliest recovery from hypoesthesia or an absence of hypoesthesia, and lower lip hypoesthesia was less with the Ob method than the ODP method.

Indoleamine 2,3-dioxygenase is a novel prognostic indicator for endometrial cancer.

Indoleamine 2,3-dioxygenase (IDO) is a tryptophan-catabolising enzyme inducing immune tolerance. The present study aimed to investigate IDO expression and its prognostic significance in endometrial cancer. Indoleamine 2,3-dioxygenase expression in endometrial cancer tissues (n = 80) was immunohistochemically scored as four groups (IDO-, 1+, 2+, and 3+). The high IDO expression (IDO2+ or 3+) in tumour cells was found in 37 (46.3%) of the 80 cases, and was positively correlated with surgical stage, myometrial invasion, lymph-vascular space involvement, and lymph node metastasis, but not with the histological grade. Patients with high IDO expression had significantly impaired overall survival and progression-free survival (PFS) (P = 0.002 and P = 0.001, respectively) compared to patients with no or weak expression of IDO (IDO- or 1+). The 5-year PFS for IDO-/1+, 2+, and 3+ were 97.7, 72.9, and 36.4%, respectively. Even in patients with early-stage disease (International Federation of Gynecology and Obstetrics I/II, n = 64), the PFS for IDO2+/3+ was significantly poor (P = 0.001) compared to that for IDO-/1+. On multivariate analysis, IDO expression was an independent prognostic factor for PFS (P = 0.020). These results indicated that the high IDO expression was involved in the progression of endometrial cancer and correlated with the impaired clinical outcome, suggesting that IDO is a novel and reliable prognostic indicator for endometrial cancer.

Angiotensin II type 1 receptor expression in ovarian cancer and its correlation with tumour angiogenesis and patient survival.

Angiotensin II, a main effector peptide in the renin-angiotensin system, acts as a growth-promoting and angiogenic factor via type 1 angiotensin II receptors (AT(1)R). We have recently demonstrated that angiotensin II enhanced tumour cell invasion and vascular endothelial growth factor (VEGF) secretion via AT1R in ovarian cancer cell lines in vitro. The aim of the present study was to determine whether AT1R expression in ovarian cancer is correlated with clinicopathological parameters, angiogenic factors and patient survival. Immunohistochemical staining for AT1R, VEGF, CD34 and proliferating cell nuclear antigen (PCNA) were analysed in ovarian cancer tissues (n = 67). Intratumour microvessel density (MVD) was analysed by counting the CD34-positive endothelial cells. Type 1 angiotensin II receptors were expressed in 85% of the cases examined, of which 55% were strongly positive. Type 1 angiotensin II receptors expression was positively correlated with VEGF expression intensity and MVD, but not with histological subtype, grade, FIGO stage or PCNA labelling index. In patients who had positive staining for AT1R, the overall survival and progression-free survival were significantly poor (P = 0.041 and 0.017, respectively) as compared to those in patients who had negative staining for AT1R, although VEGF, but not AT1R, was an independent prognostic factor on multivariate analysis. These results demonstrated that AT1R correlated with tumour angiogenesis and poor patient outcome in ovarian cancer, suggesting its clinical potential for a novel molecular target in strategies for ovarian cancer treatment.

Expression of bone morphogenetic protein-2 and proliferating cell nuclear antigen during distraction osteogenesis in the mandible in rabbits.

We examined the expression of bone morphogenetic protein-2 (BMP-2) and proliferating cell nuclear antigen (PCNA) during distraction osteogenesis in the mandible in rabbits. Twenty-four rabbits each had an osteotomy of the left mandibular body, and distraction devices were fixed. The bone was distracted at a rate of 1mm/day for 10 days. Four rabbits were killed at each of 1, 3, 7, 14, and 28 days after completion of distraction, and the mandibles examined radiographically, histologically, and immunohistochemically. Four rabbits that had not been operated on served as controls. Immunohistochemical analysis showed that BMP-2 and PCNA both appeared initially at the edge of the osteogenesis, but tended to disappear after 14 days. After 1, 3, 7, and 14 days after distraction, the ratio of stained cells was significantly higher than in the control group (p<0.05), during the period that active bone formation was shown radiographically and histologically. These results suggest that BMP-2 plays an important part in the induction of bone formation during distraction osteogenesis.

Ikaros is expressed in human extravillous trophoblasts and involved in their migration and invasion.

The transcriptional factor Ikaros was originally found to function as a key regulator of lymphocyte differentiation. In addition, we have reported that Ikaros regulates the human placental leucine aminopeptidase (P-LAP)/insulin-regulated aminopeptidase (IRAP) gene in choriocarcinoma trophoblastic cells, suggesting that Ikaros might be involved in placental development, while even its presence in human placenta remains undetermined. We therefore sought to clarify the location and roles of Ikaros in human placenta. Immunohistochemical analysis showed modest Ikaris expression in syncytium, and intense expression in extravillous trophoblasts (EVTs) in first trimester placenta. Western blot analysis showed that villous trophoblasts principally expressed Ikaros-2/3, while Ikaros-x (Ikx) was predominantly expressed in cultured EVTs. Furthermore, to investigate the functional role of Ikx in EVTs, the EVT cell line HTR-8/SVneo was infected with a retrovirus vector expressing the hemagglutinin (HA)-tagged dominant negative isoform Ikaros-6 (Ik6), which prevents the DNA-binding activity of Ikx. Antibody against HA showed successful transduction of Ik6 in HTR-8/SVneo cells on immunocytochemistry and Western blotting. Transduction of Ik6 significantly reduced the migratory and invasive abilities of HTR-8/SVneo cells. These results suggest that Ikx is involved in migration and invasion of EVTs in early placentation.

Changes in condylar long axis and skeletal stability after bilateral sagittal split ramus osteotomy with poly-L-lactic acid or titanium plate fixation.

This study was designed to assess skeletal stability after bilateral sagittal split ramus osteotomy (BSSO) and fixation with a poly-l-lactic acid (PLLA) plate, as compared to that after BSSO and fixation with a titanium plate, and to analyze the change in the condylar long axis after these procedures. The study group comprised 40 patients who had mandibular prognathism (20, titanium group; 20, PLLA group). The groups were randomized to show similar distributions of preoperative SNB. All patients underwent BSSO setback by the Obwegeser method. Fixation was done with bent titanium plates or bent PLLA plates, applied in a similar manner. Lateral, frontal, and submental-vertical cephalograms were analyzed preoperatively and postoperatively. The maximum mouth opening range and the incidence of temporomandibular disorders were also evaluated. There was no significant difference in the right condylar angle or width between the two groups, but the left condylar angle and width, gonial angle, and ramus inclination differed significantly between them (P<0.05). SNA, SNB, and ANB were similar in both groups. There was no significant difference between the groups in maximum mouth opening range or temporomandibular disorders. We conclude that the change in condylar angle after BSSO and fixation with a titanium plate is greater than that after BSSO and fixation with a PLLA plate, but skeletal stability related to the occlusion is similar for the two procedures.