Pro-apoptotic activity of inhibitory PAS domain protein (IPAS), a negative regulator of HIF-1, through binding to pro-survival Bcl-2 family proteins.

Inhibitory PAS (Per/Arnt/Sim) domain protein (IPAS) is a dominant negative transcription factor that represses hypoxia-inducible factor 1 (HIF-1) activity. In this study, we show that IPAS also functions as a pro-apoptotic protein through binding to pro-survival Bcl-2 family members. In a previous paper, we reported that NF-κB-dependent IPAS induction by cobalt chloride repressed the hypoxic response in PC12 cells. We found that prolonged incubation under the same conditions caused apoptosis in PC12 cells. Repression of IPAS induction protected cells from apoptosis. Furthermore, knockdown of IPAS recovered cell viability. EGFP-IPAS protein was localized in both the nucleus and the cytoplasm, with a large fraction associated with mitochondria. Mitochondrial IPAS induced mitochondria depolarization and caspase-3 activation. Immunoprecipitation assays revealed that IPAS is associated with Bcl-x(L), Bcl-w and Mcl-1. The association of IPAS with Bcl-x(L) was also observed in living cells by the FLIM-based FRET analysis, indicating direct binding between the two proteins. IPAS contributed to dysfunction of Bcl-x(L) by inhibiting the interaction of Bcl-x(L) with Bax. These results demonstrate that IPAS functions as a dual function protein involved in transcription repression and apoptosis.

Observation of an energetic radiation burst from mountain-top thunderclouds.

During thunderstorms on 20 September 2008, a simultaneous detection of gamma rays and electrons was made at a mountain observatory in Japan located 2770 m above sea level. Both emissions, lasting 90 sec, were associated with thunderclouds rather than lightning. The photon spectrum, extending to 10 MeV, can be interpreted as consisting of bremsstrahlung gamma rays arriving from a source which is 60-130 m in distance at 90% confidence level. The observed electrons are likely to be dominated by a primary population escaping from an acceleration region in the clouds.

Pattern classification of nevus with texture analysis.

The purpose of this research is to classify the pattern on the surface of the nevus. The digital image that contains one nevus is classified into three kinds of patterns of homogeneous pattern, globular pattern, and reticular pattern by the texture analysis. The tumor part in the image is specified first, and the specified tumor part is divided into some sub-images. Afterwards, the amount of the texture features of each sub-image was calculated. The pattern was classified by the discriminant analysis based on the amount of the texture features. As a result, the patterns could be classified correctly into three categories at the ratio of 94%.

The prevalence of IgE sensitization to formaldehyde in asthmatic children.

BACKGROUND: Formaldehyde (FA) is well documented as a cause of occupational asthma. Recently, attention has been paid to FA as an allergen and a pollutant that enhances allergic sensitization. We have investigated the prevalence of FA-specific IgE in asthmatic children and the correlation between IgE sensitization to FA and the severity of asthma. METHODS: One hundred and fifty-five children were investigated, 122 of them asthmatic and 33 nonallergic. Specific IgE against FA was measured by CAP RAST. In addition, the patients answered a questionnaire, containing questions on clinical features of their asthma, their living conditions, and symptoms of mucosal irritation. RESULTS: Of all the subjects assessed, FA-specific IgE was detected in only two asthmatics, and their IgE levels of FA were low (0.42 and 0.46 UA/ml). One of the two patients with FA-specific IgE had severe asthma and frequent symptoms of mucosal irritation, but the other had mild asthma and only rare symptoms of mucosal irritation. CONCLUSIONS: The prevalence of IgE sensitization to FA appears very low in Japanese children, whether or not they have asthma. Therefore, it appears likely that FA is not one of the major allergens causing childhood asthma.

Pedicle bone grafting versus transtrochanteric rotational osteotomy for avascular necrosis of the femoral head.

Segmental collapse occurs in the early stage of avascular necrosis (AVN) of the femoral head, and is associated with a poor prognosis. Since it develops at a relatively young age, the long-term outcome after total hip replacement is a major concern. We have compared the long-term results of pedicle bone grafting (PBG) with those of transtrochanteric rotational osteotomy (TRO). In the PBG group there were 23 men (27 hips) and three women (4 hips) with a mean age at the time of surgery of 38 years and a mean follow-up of 13 years. In the TRO group there were 44 men (55 hips) and 19 women (22 hips) with a mean age at the time of surgery of 39 years and a mean follow-up of seven years. Failure was defined as a need for total hip replacement or a Harris hip score below 70. The long-term results were similar for the two groups. The survival rates at five and ten years were 85% and 67%, respectively, in the PBG group, and 71% and 61%, respectively, in the TRO group, according to Kaplan-Meier survivorship analysis. In the TRO group patients in stage II had significantly better results that those in stage III.

TUCAN, an antiapoptotic caspase-associated recruitment domain family protein overexpressed in cancer.

Caspase-associated recruitment domains (CARDs) are protein interaction domains that participate in activation or suppression of CARD-carrying members of the caspase family of apoptosis-inducing proteases. A novel CARD-containing protein was identified that is overexpressed in some types of cancer and that binds and suppresses activation of procaspase-9, which we term TUCAN (tumor-up-regulated CARD-containing antagonist of caspase nine). The CARD domain of TUCAN selectively binds itself and procaspase-9. TUCAN interferes with binding of Apaf1 to procaspase-9 and suppresses caspase activation induced by the Apaf1 activator, cytochrome c. Overexpression of TUCAN in cells by stable or transient transfection inhibits apoptosis and caspase activation induced by Apaf1/caspase-9-dependent stimuli, including Bax, VP16, and staurosporine, but not by Apaf1/caspase-9-independent stimuli, Fas and granzyme B. High levels of endogenous TUCAN protein were detected in several tumor cell lines and in colon cancer specimens, correlating with shorter patient survival. Thus, TUCAN represents a new member of the CARD family that selectively suppresses apoptosis induced via the mitochondrial pathway for caspase activation.

Pedicle myocutaneous flaps for reconstruction following total pelvic exenteration of intrapelvic recurrent rectal cancer: report of a case.

A vast metastatic tumor mass of recurrent rectal carcinoma in the intrapelvic organs is commonly considered unsuitable for total pelvic exenteration (TPE); first, because it is unlikely that it would improve the prognosis and health-related quality of life of the patient, and second, because of the difficulties involved in this surgical technique. However, by using a plastic surgery technique involving reconstruction by filling the pelvic dead space with pedicle myocutaneous flaps (PMF), primary wound closure and extensive resection of the perineum can be achieved, whereby postoperative metastasis may be prevented. We report herein the case of a 71-year-old man found to have local recurrence in the perineum with extensive invasion of the soft tissue as well as adjacent organs, 20 months after abdominal perineal resection for rectal carcinoma. TPE with extensive resection of the perineal soft tissue was performed, followed by perineal reconstruction and packing of the pelvic dead space with PMF, mainly constructed from the gracilis and sartorius muscles of both femurs. His postoperative course was uneventful and he has remained free of local recurrence and symptomatic perineal complaints for 1 year. In this report, we examine the effectiveness of using the gracilis muscle for PMF in intrapelvic and perineal reconstruction after TPE.

External venous shunt as a solution to venous thrombosis in microvascular surgery.

We report a case in which venous occlusion occurred after fibular osteocutaneous flap transfer for open tibial fracture. The flap was salvaged using an external venous shunt. The shunt consisted of a Teflon catheter for intravenous infusion, a three-way stopcock, a urokinase-immobilised catheter and a pressurising bag for arterial-pressure monitoring. This method can be used to establish venous drainage when there is no accessible recipient vein. It may improve the success rate of microvascular surgery and extend the indications for surgery.

Endoscopic correction of pectus excavatum.

Endoscopic surgery is minimally invasive and can be used to achieve superior cosmetic results. Conventional correction of pectus excavatum results in a long scar. Correction by use of endoscopic surgery involves a smaller skin incision. In this study, endoscopic correction of pectus excavatum was performed in 20 cases. A small transverse skin incision was made above the xyphoid process. A wide area beneath the pectoralis major muscle was dissected under endoscopic visualization. Subperichondrial resection was performed under direct visualization when possible. Subperichondrial resection of the third or fourth rib was performed under endoscopic visualization. Ravitch's chondrotomy of the second or third rib was performed under endoscopic visualization. Endoscopy was also useful for sternal elevation, with minimal risk of pleural perforation. Kirschner wire was inserted percutaneously under the sternum to prevent postoperative paradoxical respiration. In all cases, the postoperative course was uneventful. The advantages of endoscopic pectus excavatum correction are a short scar, control of bleeding, safe dissection of the pleura from the sternum without the risk of pleural perforation, and ease of sternal elevation without injury to the intramammary vessels. However, the endoscopic operation is long and is not useful in adults because subperichondrial resection in adults is difficult to perform.

Apoptotic protease activating factor 1 (Apaf-1)-independent cell death suppression by Bcl-2.

Reportedly, antiapoptotic Bcl-2 family proteins suppress apoptosis by binding to and inhibiting members of the CED-4 family of caspase activators. To explore this question, we used embryonic stem (ES) cells in which one (-/+) or both (-/-) copies of the gene encoding apoptotic protease activating factor 1 (Apaf-1), a CED-4 homologue, were disrupted by homologous recombination. Stable clones of heterozygous (-/+) and homozygous (-/-) Apaf-1 knockout ES cells that overexpressed Bcl-2 were generated. Withdrawal of serum growth factors or stimulation of heterozygous ES cells with staurosporine (STS), ultraviolet (UV)B irradiation, etoposide (VP16), or cisplatin induced apoptosis followed by cell death (determined by failure to exclude propidium iodide dye). These cell death stimuli also induced activation of several types of caspases and loss of mitochondrial membrane potential (DeltaPsi) in heterozygous (+/-) Apaf-1 knockout ES cells. In addition, overexpression of Bcl-2 protected against these events in Apaf-1-expressing ES cells. In contrast, STS, UVB, and VP16 induced little or no caspase activation and apoptosis in homozygous (-/-) Apaf-1 knockout ES cells. Nevertheless, Apaf-1-deficient ES cells subjected to these cell death stimuli or deprived of growth factors did eventually die through a nonapoptotic mechanism associated with loss of DeltaPsi. Moreover, Bcl-2 overprotection preserved DeltaPsi, reduced the percentage of Apaf-1(-/)- ES cells undergoing cell death, and increased clonigenic survival. The extent of Bcl-2-mediated cytoprotection was not significantly different for heterozygous (-/+) versus homozygous (-/-) Apaf-1 knockout cells. Furthermore, although Bcl-2 could be readily coimmunoprecipitated with Bax, associations with Apaf-1 were undetectable under conditions where Apaf-1 interactions with procaspase-9 were observed. We conclude that Bcl-2 has cytoprotective functions independent of Apaf-1, preserving mitochondrial function through a caspase-independent mechanism.

Ulnar artery graft for myocardial revascularization.

We present a 60-year-old man who underwent coronary artery bypass grafting using an ulnar artery as one of the grafts intended to release angina pectoris. Previously, his right leg had been amputated following a traffic accident. The blood supply of his left leg was reduced due to atherosclerotic stenotic change (left ankle pressure index 0.6). He had been under treatment for severe diabetes mellitus for 4 years. Coronary angiography revealed severe stenosis in the triple coronary artery system. Immediate myocardial revascularization was considered necessary. We considered that saphenous vein grafts and bilateral internal thoracic artery grafts were unsuitable for this patient. Moreover, Allen's test was positive in the bilateral forearms. Coronary artery bypass surgery consisted of left internal thoracic artery grafting to the left anterior descending artery, right gastroepiploic artery grafting to the right coronary artery, and left ulnar artery grafting to the diagonal branch. No myocardial or hand complications were observed after surgery. Following a review of the Japanese literature, we conclude that our case is the first report of an ulnar artery graft for coronary artery bypass grafting in Japan.

Hypoglycaemic brain lesions in a dog with insulinoma.

A 5-year-old female Collie dog showed excessive salivation, vomiting and neurological signs, including hind-limb weakness, mental dullness and subsequent recumbency with paddling movements of the limbs. Blood glucose and insulin concentrations were 35 mg/dl and 70.0 microU/ml, respectively. At necropsy, two masses, one at the caudal edge of the pancreas and the other in the omentum, were found and diagnosed as insulinoma. Histological examination of the brain showed early signs of acute neuronal necrosis exclusively in the superficial layers of the cerebral cortex, in addition to spongy changes in the dentate gyrus of the hippocampus. The light microscopical findings were identical in character and distribution with those of naturally occurring hypoglycaemia in humans and experimentally induced hypoglycaemia in animals.

Natural skin reduction and breast recovery using a tissue expander after enucleation of a giant breast tumour.

We report a new use of the tissue expander for reshaping a breast after resection of a giant tumour. After resection of giant fibroadenomas, two patients had expanders inserted into the tissue defect and gradually reduced in size over five months. This facilitated healing and natural skin shrinkage and resulted in a natural shape and size.

Heterogeneous reactivity of murine epidermal Langerhans cells after application of FITC: a histochemical evaluation.

To elucidate the detailed kinetics of epidermal Langerhans cells after topical contact sensitizer stimulation, we examined ATPase or Ia positive epidermal cells of BALB/c mice in a time-spaced manner after the topical application of fluorescein isothiocyanate (FITC). We also performed double labeling of Langerhans cells in epidermal sheets with ATPase activity and Ia antigen or costimulatory molecules (B7-1 and B7-2) after the same stimulation. Observations showed that the density of ATPase positive cells and Ia positive cells decreased following a different time course; the former reached a nadir (77.4% of control) at 4 h but the latter reached a minimum (82.8% of control) at 16 h after the application of FITC. A double labeling technique revealed an increase in Ia single positive cells at 4 h as opposed to that of ATPase single positive cells at 16 h after application. Both costimulatory molecules were expressed on the dendritic processes of many Langerhans cells as a dotty pattern at 4 h after application; B7 positive and ATPase negative areas were observed at this time. On electron microscopic observation, a few activated Langerhans cells found in the dermis at 4 h after application had distinctive profiles compared with residual Langerhans cells in the epidermis. These findings suggest that there is a heterogeneity of reactivity to FITC in epidermal Langerhans cells, and that only a small portion of them migrates from the epidermis during sensitization. The findings also indicate the importance of the interaction between the Langerhans cell and its surrounding microenvironment in the epidermis for its activation. In addition, the results indicate that the enzymatic and the phenotypic markers do not definitively reflect the presence (or absence) of Langerhans cells.

Human Daxx regulates Fas-induced apoptosis from nuclear PML oncogenic domains (PODs).

Daxx was first identified as a protein that binds the cytosolic domain of Fas and links this receptor to an apoptosis pathway involving activation of Jun N-terminal kinase (JNK). We show here that cells overexpressing the human homolog of Daxx (hDaxx) display enhanced sensitivity to apoptosis induced by Fas but not by several other cell death stimuli. hDaxx-mediated enhancement of Fas-induced apoptosis was correlated with accelerated activation of caspases but not with JNK induction. Although specifically enhancing Fas function, hDaxx does not bind Fas and instead is found in the nucleus where it localizes to PML oncogenic domains (PODs). Moreover, the hDaxx protein also exhibits the ability to repress transcription. Mutagenesis studies demonstrated a correlation between the localization of hDaxx to PODs and its ability to enhance Fas-induced cell death. Arsenic trioxide (As(2)O(3)), an agent that accentuates POD formation, collaborated synergistically with overexpression of hDaxx to increase cellular sensitivity to Fas-induced apoptosis. Taken together, these findings argue that hDaxx promotes sensitivity to Fas from a nuclear location, probably by modulating the transcription of genes involved in Fas-induced caspase activation and apoptosis.

Successful surgical treatment of a solitary parapharyngeal metastasis from thyroid cancer, using the mandibular swing-transcervical approach: report of a case.

A 72-year-old man presented with a right parapharyngeal mass, 4 cm in diameter, which was subsequently diagnosed as a metastasis originating from papillary carcinoma of the thyroid gland. The parapharyngeal tumor was successfully removed by the mandibular swing-transcervical approach with pharyngeal reconstruction, performed using a buccal mucosal island flap based on the facial artery. His postoperative course was uneventful, and the preoperative clinical symptoms such as dysphagia and headaches completely resolved after surgery.

The lumbar artery perforator based island flap: anatomical study and case reports.

A lumbar artery island flap can be elevated based on a single lumbar artery. We studied the vascular anatomy using 21 specimens of lumbar arteries in 11 cadavers, and investigated the skin territory of the artery using fluorescein injection. We observed lumbar perforators emerging through the lumbar fascia at the lateral border of the erector spinae muscle, situated 5-9 cm from the midline. The diameter of the vascular bundle at the site of perforation ranged from 1 to 5 mm. Perforators of the second and fourth lumbar arteries were much more developed than others. The cutaneous territory supplied by the second lumbar artery extended from the posterior midline to the lateral border of the rectus sheath, and at least 10 cm above the anterosuperior iliac spine. We transferred four clinical flaps for coverage of ulcers on the lower back. All flaps survived and their donor site defects were closed primarily. The cadaver dissection, the injection study and our clinical success have confirmed the feasibility of lumbar artery island flaps.

Endoscopic omental harvest.

An omental flap is useful in reconstructive surgery, but harvesting such a flap generally requires laparotomy. However, endoscopic surgery facilitates harvesting an omental flap without open laparotomy. We performed endoscopic omental harvest in two patients. We described the procedure of endoscopic omental harvest, which is different from that reported previously. Four access ports were required: two placed lateral to the right rectus margin, one placed lateral to the left rectus margin, and one placed in the infraumbilical area. The stomach was suspended from the peritoneum for the dissection of the gastric rami. The vessels from the gastroepiploic arcade to the greater curvature of the stomach were individually clipped and divided. The omentum then was dissected to the transverse colon and the lower portion of the omentum was dissected along the transverse colon. Finally, the right side of the omentum was dissected. The omentum was transferred using the right gastroepiploic vessels for anastomosis. The advantages of endoscopic harvest are an inconspicuous scar, minimal operative pain, and early recovery. The disadvantages include a long procedure time. At the present time, endoscopic harvest of the left side of the omentum is problematic because of difficulty in identifying the left omental artery and the risk of injury to the spleen. However, these limitations will likely be resolved in the future.