FOXO3 is essential for CD44 expression in pancreatic cancer cells.

Pancreatic ductal adenocarcinoma (PDAC) is one of the most fatal types of cancer and the 5-year survival rate is only 5%. Several studies have suggested that cancer stem cells (CSCs) are thought to be involved in recurrence and metastasis and so it is essential to establish an approach targeting CSCs. Here we have demonstrated that cyclic guanosine monophosphate (cGMP) suppressed CD44 expression and the properties of CSCs in PDAC. Microarray analysis suggested that cGMP inhibited Forkhead box O3 (FOXO3), which is known as a tumor suppressor. Surprisingly, our data demonstrated that FOXO3 is essential for CD44 expression and the properties of CSCs. Our data also indicated that patients with high FOXO3 activation signatures had poor prognoses. This evidence suggested that cGMP induction and FOXO3 inhibition could be ideal candidates for pancreatic CSC.

Successful treatment of nonunion with teriparatide after failed ankle arthrodesis for Charcot arthropathy.

We describe a case of successful treatment to nonunion after multiple arthrodesis operations for Charcot arthropathy with teriparatide. We describe the case of a 25-year-old woman with severe Type I diabetes mellitus that resulted in nonunion after multiple arthrodesis operations for Charcot arthropathy. The woman sustained a femoral shaft fracture for which she underwent surgery with intramedullary nail fixation. Immediately after surgery, an empiric course of teriparatide was initiated. Femoral shaft fracture healing was observed after 2 weeks, and the woman was able to walk 12 weeks after the surgery, at which point plain film and computed tomography images revealed complete union of the ankle.

Generation of dendritic cells and macrophages from human induced pluripotent stem cells aiming at cell therapy.

This report describes generation of dendritic cells (DCs) and macrophages from human induced pluripotent stem (iPS) cells. iPS cell-derived DC (iPS-DC) exhibited the morphology of typical DC and function of T-cell stimulation and antigen presentation. iPS-DC loaded with cytomegalovirus (CMV) peptide induced vigorous expansion of CMV-specific autologous CD8+ T cells. Macrophages (iPS-MP) with activity of zymosan phagocytosis and C5a-induced chemotaxis were also generated from iPS cells. Genetically modified iPS-MPs were generated by the introduction of expression vectors into undifferentiated iPS cells, isolation of transfectant iPS cell clone and subsequent differentiation. By this procedure, we generated iPS-MP expressing a membrane-bound form of single chain antibody (scFv) specific to amyloid ß (Aß), the causal protein of Alzheimer's disease. The scFv-transfectant iPS-MP exhibited efficient Aß-specific phagocytosis activity. iPS-MP expressing CD20-specific scFv engulfed and killed BALL-1 B-cell leukemia cells. Anti-BALL-1 effect of iPS-MP in vivo was demonstrated in a xeno-transplantation model using severe combined immunodeficient mice. In addition, we established a xeno-free culture protocol to generate iPS-DC and iPS-MP. Collectively, we demonstrated the possibility of application of iPS-DC and macrophages to cell therapy.

Reversed lateral upper arm flap with a vascularised fragment of the humerus for reconstruction of ulna shaft fracture after resection of malignant tumour: a case report.

The lateral upper arm flap (LAF) is used for a pedicled flap with the vascular arcade at the elbow. There has been no report, however, with regard to the reverse lateral upper arm flap (R-LAF) with a vascularised humerus as an osteocutaneous flap. We describe the case of a 72-year-old woman with a fracture of the ulna after wide resection of a malignant tumour at the proximal half of the forearm, which was reconstructed with an R-LAF with a vascularised fragment of the distal humerus. The flap has successfully survived and the fracture was healed without any adverse event. The R-LAF with vascularised bone is a useful option for the simultaneous treatment of soft tissue and bone defects.

A randomized comparison of locking and non-locking palmar plating for unstable Colles' fractures in the elderly.

This study compared the effectiveness of locking and non-locking palmar plating for unstable Colles' fractures in the elderly. The patients treated with locking plates included 4 men and 18 women with a mean age of 68 years (Group A) and those treated with non-locking plates included 3 men and 28 women with a mean age of 74 years (Group B). Radiographic parameters, including palmar tilt, radial inclination and radial length were measured before surgery, after surgery and at final followup. There were no significant differences in respect of any of the radiographic parameters between the two groups pre-operatively. After surgery, all of the radiographic parameters were improved in both groups and there were no significant differences between the two groups at final followup.

[Differences between gynecological cancer patients taken care for at home and those taken care for at the hospital].

Thirty-five patients with end-stage gynecological cancer, twelve taken care for at home care and 23 taken care for at our hospital, were compared. The life span after recurrence was significantly (p < 0.05) longer in the group taken care for at home. The number of persons living with the patient was also significantly (p < 0.01) larger in the group taken care for at home. Moreover, the hospitalization in the end stage of the disease was significantly (p < 0.01) shorter in the group taken care for at home. The average hospitalization in the end was for 6.5 days. Four patients died at their home. Average period the 12 patients stayed at home was 52.7 days. Following matters are important in order to provide home care. 1. We must know the biological characteristics of the cancer. 2. It is important that it does not lose the timing to which shift to the home care, keeping good doctor-patient relationship. 3. Limit of the therapy against cancer and importance of treatment at home should be carefully and clearly explained to the patient.

Isolation and characterization of the gene encoding mouse tax-responsive element-binding protein (TREB) 5.

TREB5/hXBP-1/HTF is a basic region leucine zipper protein which binds to a cyclic AMP responsive element (CRE)-like element in both human T-cell leukemia virus type 1 and human major histocompatibility complex (MHC) class II genes. To analyze the structure and transcription regulation of the TREB5 gene, we isolated the mouse TREB5 gene and cDNA. The mouse TREB5 gene contains five exons and four introns and spans approximately 5 kb. The deduced amino acid sequence of mouse TREB5 exhibited 77% and 94% homology to human and rat TREB5, respectively. The b-zip structure is completely conserved in mouse, rat and human. Southern blot analysis of the mouse genomic DNA demonstrated that positive bands exactly coincide with those expected from sequences of the cloned genes, indicating that the mouse TREB5 gene is present as a single copy. The transcription start site of the mouse TREB5 gene was mapped to -15 bp upstream from the ATG initiation codon. Promoter analysis of serial deletion mutants revealed that the -142 bp upstream region is the minimum sequence to promote mouse TREB5 gene expression and that the -1.0 kb upstream region is required for full promoter activity.

Absorption of calcium, magnesium, phosphorus, iron and zinc in growing male rats fed diets containing either phytate-free soybean protein or soybean protein isolate or casein.

The effect of dietary phytate-free soybean protein (PFS) on intestinal mineral absorption and retention was examined in growing male rats using a three-day mineral balance technique. The rats were fed diets containing PFS, soybean protein isolate (SPI) or casein at a 20% level for 5 wk. Total calcium (Ca), magnesium (Mg), phosphorus (P), iron (Fe) and zinc (Zn) contents in diets were adjusted to 0.35, 0.05, 0.7, 0.0035 and 0.003%, respectively, by supplementation of the diet with their salts. Mineral absorption and retention ratios in rats fed the PFS diet were significantly higher than those in rats fed either the SPI or casein diet. These results suggest that PFS may be a promising dietary protein source for improving the mineral bioavailability in humans.

Antioxidant activity of soya hypocotyl tea in humans.

Antioxidative activity of isoflavones has not been shown in humans. Newly-developed isoflavone-rich soya hypocotyl tea contains about 12 mg isoflavones per liter. 15 tea drinkers and 23 control young female students were randomly selected from volunteers, and underwent physical examination, blood chemistry and urinary analysis before and after one month of tea drinking. A three-day dietary record was taken before each physical examination. The tea drinkers showed a lower level of phosphatidylcholine hydroperoxide (PCOOH) and phosphatidyl-ethanolamine hydroperoxide (PEOOH) in the red blood cells and a significant reduction of 8-hydroxydeoxyguanine (8ohdG) in the urine compared to the controls.

Serum insulin-like growth factor-I, insulin-like growth factor binding protein-3, sex steroids, osteocalcin and bone mineral density in male and female rats.

Although it has been reported that the rate of weight gain and linear growth increases markedly during puberty in rats, little is known about the relationship between endocrine changes and bone mineral density (BMD) changes upon sexual maturation in these animals. The aim of this study was to examine the levels of serum insulin-like growth factor-I (IGF-I), IGF binding protein (IGFBP)-3, sex steroids and osteocalcin, and the changes in BMD in normal aging male and female rats. Male rats exhibited increases in serum IGF-I and IGFBP-3 concentrations before increases in serum testosterone levels. IGF-I and testosterone peaked at 9 weeks of age, and thereafter remained in a steady state, whereas IGFBP-3 reached a peak at 7 weeks of age, and then gradually declined. A strong correlation between serum IGF-I and IGFBP-3 levels was found in subjects 3-9 weeks old. A highly significant correlation between serum IGF-I and testosterone levels was also found. In females, serum 17 beta-estradiol, IGF-I and IGFBP-3 levels increased gradually from 3 to 5 weeks old, peaked at 9 weeks, and then decreased slowly thereafter. The correlation coefficient between serum IGF-I and IGFBP-3 was highly significant. The correlation coefficient between serum IGF-I and 17 beta-estradiol levels was weak, although it was strongest when the subjects were 3-9 weeks old. Serum osteocalcin is a marker of bone formation; its level remained relatively high from 3 to 9 and from 3 to 7 weeks of age in males and females, respectively, although osteocalcin in both sexes declined gradually with age. As for bone mass, sharp increases in BMD in the tibia, femur and lumbar vertebrae appeared earlier in female than in male rats, and the BMD in females tended to be higher than in males between 5 and 9 weeks old. After 9 weeks of age, BMD in males was higher than that in females, as BMD in males continued to increase whereas females tended to remain in a steady state after this stage. The correlation coefficients between tibial BMD and serum IGF-I or IGFBP-3 levels were highly significant when the subjects were from 3 to 9 weeks old. Taken together, these results suggest that BMD development occurs earlier in female than in male rats. This sex-related difference in changes in the BMD pattern may result from the earlier onset of puberty in females, and from sex-specific differences in concentrations of IGF-I, IGFBP-3 and sex steroids during maturation.

Effect of smoking on the prevalence of albuminuria in Japanese men with non-insulin-dependent diabetes mellitus.

Smoking is a risk factor for diabetic nephropathy in patients with IDDM and potentially those with NIDDM. We investigated the relationship between renal involvement and cigarette smoking in 148 men with NIDDM. The presence of renal involvement was assessed by determining the overnight urinary albumin/creatinine ratio (mg/g, ACR). The patients were divided into three groups, normo-, micro-, and macroalbuminuria, based on the ACR (< 30, 30-300, and 300 < or = mg/g, respectively). The incidence of micro-/macroalbuminuria in 81 smokers was significantly higher than that in 21 ex-smokers (stopped smoking at least 10 years prior to the study) or 40 non-smokers (53.1, 33.3, and 20.0%, respectively). The prevalence of smoking in the groups of patients with normo-, micro-, and macroalbuminuria were 45, 73, and 76%, respectively. The relative risk (odds ratio) for the prevalence of micro-/macroalbuminuria associated with smoking was 4.5 (95% CI, 1.9-11.6, P < 0.001) in smokers and was 2.0 (not significant) in ex-smokers. Our results indicate that stricter counselling about the importance of quitting smoking will be necessary in patients with NIDDM to protect against the development of diabetic nephropathy.

Changes in CA602 and CA546 concentrations in patients during in-vitro fertilization and embryo transfer, and ovarian hyperstimulation syndrome.

Changes in CA602 and CA546, two carbohydrate-related antigens newly established in our institute, were evaluated during the course of in-vitro fertilization and embryo transfer. The concentration of CA602, an antigen with characteristics similar to those of CA125, did not change substantially until embryo transfer. In patients with clinical evidence of ovarian hyperstimulation syndrome (OHSS), a significant increase in serum concentrations of CA602 was observed 10 days after embryo transfer (P < 0.01). There were no significant changes in the patients without OHSS. A significant elevation of oestradiol concentration in OHSS patients was also observed, with the variation before oocyte retrieval being greater than that seen with CA602. The concentration of CA546, which had lower false-positive rates in benign disease, did not differ significantly in the OHSS and non-OHSS patients. Our data demonstrate that serum concentrations of CA602 are increased even in early stages of OHSS, and suggest a possible role for this antigen as a marker for OHSS.

Diffuse lipomatosis: imaging features.

A case of diffuse lipomatosis is reported. A 54-year-old man presented with a large mass in the left gluteal region. Plain radiographs showed hyperostosis of the phalanx of the left third toe. The diagnosis was aided by magnetic resonance imaging and computed tomography which clearly demonstrated a diffuse overgrowth of fatty tissue in the left buttock and the extremity involving the subcutaneous and gluteal muscular tissue.

Functional evaluation of flail hip joint after periacetabular resection of the pelvis.

Five patients subjected to flail hip joint after resection of primary bone tumor of the pelvis involving acetabular region were examined with respect to function. When a thick portion of the supraacetabular pelvic neck was left in place, the operated legs functioned well regardless of whether the head of the femur was placed anterior or posterior to the iliac wing. In a patient who had only a thin portion of the iliac wing left in place and in those who underwent total excision of the ilium or hemiresection of the pelvis, the function of operated legs was poorer than with a thick portion of the supraacetabular pelvic neck left in place, but still more than 50% of leg function remained. Although flail hip joint results in a larger leg-length discrepancy than do other techniques, it enables favorable healing of the operative wound. Therefore, this method should be considered for women or sedentary patients with primary bone tumor of the pelvis involving acetabular region.

Trypsin inhibitor activity in commercial soybean products in Japan.

The effect of trypsin inhibitor (TI) in soybean on the prevention of cancer has been reported in animal studies. However, the actual intake of active TI in daily life is not known. In this experiment we measured TI activity in various soybean products and the effects of some factors of processing on the TI activity. Average TI activity in whole soybean was 4,819 U/100 g, and the average percentages of remaining activity in products against that of whole soybean were momen-tofu 2.5%, yose-tofu 3.4%, kinugoshi-tofu 4.3%, jyuten-tofu 7.9%, soymilk 13.0%, natto 0.7%, soy sauce 0.8% and miso 0.3%. By heating soymilk at 100 degrees C, TI activity decreased to 11% at 10 min and 5% at 20 min. After heating soymilk in a water-bath for 15 min at 75 and 100 degrees C, TI activity decreased to 35 and 12%, respectively. The TI activity of tofu was proportional to the remaining whey. The effects of chemicals used for the coagulation of soy protein and foam-removal on TI activity were little. The results suggest that soybean products retain 2.5-12.5% TI activity of the whole soybean and that humans are consuming some active TI in their daily lives.

[Increased urinary iron excretion rate in patients with non-insulin dependent diabetes mellitus].

Urinary iron excretion rate(u-FeER, microgram/min) in urine at night time from 58 patients with non-insulin dependent diabetes mellitus and 8 controls was measured by atomic absorption. The patients were divided into three groups according to urinary albumin excretion rate(u-AER), namely, Group I(n = 44): less than 20 micrograms/min of u-AER; Group II(n = 9): 20 < or = u-AER < 200 micrograms/min; Group III(n = 5): more than 200 micrograms/min of u-AER. The u-FeER in group III(56.3 +/- 14.8 ng/min, mean +/- SEM) was significantly higher than that in controls (4.0 +/- 1.6ng/min), group I(8.3 +/- 1.6 ng/min) and group II(18.5 +/- 6.5ng/min). The increase of u-FeER in group III indicates that urinary iron may at least partly play a role in development of diabetic nephropathy.

[Applications of tumor markers to the screening of endometrial and ovarian cancers].

Several new trials, using tumor markers in the screening for gynecological malignancies, have been conducted. For assisting the cytological diagnosis of uterine endometrial cancers, the new EIA method using cytological specimens and the monoclonal antibody against endometrial cancer cells, MSN-1, was developed. This method could help to discriminate between cancer and normal cells, so this would result in decrease the numbers of suspicious cases on cytological diagnosis. For ovarian cancers, especially to identify high-risk groups, two carbohydrate-related antigens, CA602 and CA546, were employed. The combined use of these two markers showed a high potentiality to detect ovarian cancers. GAT (galactosyltransferase associated with tumor), an isoform of galactosyltransferase, could rescue the false-positive cases with endometriotic cysts. These new methods with tumor markers are supposed to be handy tools in the screening for gynecological malignancies.