Low-dose hyperthermia enhances the antitumor effects of chemotherapy in squamous cell carcinoma.

Esophageal squamous cell carcinoma is a highly aggressive neoplasm and the sixth leading cause of global cancer-related death; the 5-year survival rate for esophageal cancer is only about 20%-25% for all stages. Therefore, improving the therapeutic effect is important. This study assessed whether low-dose hyperthermia (LDH) enhances the antitumor effects of chemotherapy. The antitumor effect of chemotherapy with/without LDH in the squamous cell carcinoma cell line SCCVII was evaluated. A comprehensive analysis was performed with real-time polymerase chain reaction (PCR) to study the hyperthermia-induced changes in the gene expression of SCCVII cell lines. In addition, the cytotoxic and apoptotic changes in the cells treated with LDH combined with/without 5-fluorouracil (5-FU) were measured. LDH combined with 5-FU (10 nM) strongly inhibited the cell growth of SCCVII, with flow cytometry showing an increased population of apoptotic cells. PCR showed that LDH promoted a 25.22-fold increase of p53 mRNA and 18.08-fold increase of Bax mRNA in vitro. MDR1 expression was decreased to 28.7% after LDH. This treatment can result in much higher efficacy of antitumor drugs. After LDH, the expressions of TS decreased to 12.06%, OPRT increased by 4.17-fold, and DPD did not change (1.03-fold). This transformations will induce susceptibility to 5-FU. LDH may be a useful enhancer of chemotherapy drugs for squamous cell carcinoma.

Modern precipitation of hydrogenetic ferromanganese minerals during on-site 15-year exposure tests.

Redox-sensitive metallic elements, Mn and Fe, are oxidized in deep sea waters and form abundant ferromanganese crusts and nodules on the world's ocean floors at ultraslow rates of growth. This process of oxidation and the mechanism of precipitation are yet unknown. In this paper, the results of the first successful, long-term, on-site experiment of mineral precipitation that ascertains modern, ongoing hydrogenetic deposition of oxide materials from normal seawaters at water depths of 900-4500 m of geologically active and inactive environments are presented. We succeeded in the in-situ precipitation experiment on the sea floor and characterized the precipitates using high-resolution and submicron-scale chemical, mineralogical, and structural analyses. The installed artificial plates of glass, ceramics, and plastic yielded spread-out particles of sizes varying from one to a few micrometers in diameter, of coccoid-like irregular shapes, with a maximum of 1,000-10,000 individual particles/mm2/year after 12-15 years of exposure. The results indicated a continuous substantial growth of the hydrogenetic minerals if both Mn and Fe are supplied to the bottom waters. The mineralogical, chemical, and structural properties of the precipitates are similar to those of the natural precipitates on the seabed that are made up of hydrogenetic ferromanganese crusts and nodules, together with settling sediments, suspended hydrothermal particles, or microbial precipitates from cultivated Mn-oxidizing bacteria. Our work presents new realistic insight into proposed genetic models of marine hydrogenetic ferromanganese deposits in modern diverse ocean environments.

[The strategy for aortic root reconstruction; Bentall operation or native valve preserving operation?].

Bentall operation has been a gold-standard procedure for the aortic root reconstruction in the patients with aortic root aneurysms and aortic valve insufficiency (AI). Recently, native aortic valve preserving operation has been widely spread. In this study, we described the results of aortic root reconstruction in our institution and evaluated our operative methods. We selected Bentall operation for severely damaged or cardiac redo cases, and David operation for simple annuloaortic ectasia and AI. Recently, we have utilized Valsalva graft which provides the advantage of capability of larger prosthetic valve implantation and better hemodynamic performance.

The gene poxn controls different steps of the formation of chemosensory organs in Drosophila.

The gene poxn codes for a transcriptional regulator that specifies poly-innervated (chemosensory), as opposed to mono-innervated (mechanosensory), organs in Drosophila. The ectopic expression of poxn during metamorphosis results in a transformation of the morphology and central projection of adult mechanosensory organs toward those of chemosensory organs. Here we show, by electron microscopy analysis of normal and transformed bristles and by Dil labeling of the innervating neurons, that poxn also controls the number of neurons. To determine whether poxn can transform not only the sense organ precursor cells but also their daughter cells, we examine the effects of the ectopic expression of poxn at different stages of the lineage, and we conclude that poxn can act at a late stage to affect the fate of the undifferentiated neuron.

[Coronary artery anastomosis using a cloth graft-holder attached to a stabilizer].

Numerous assist devices are usually used for coronary bypass grafting but they make the surgical field cluttered. This study proposes the use of a simple anastomotic method using a cloth graft-holder, which can be attached to a stabilizer. A graft is held between a double-folded cloth of a graft-holder and the graft-holder is attached to the stabilizer using double-stick tape, which provides steady positioning of the graft even in an up-right position. The graft can be positioned parallel with a coronary artery on either side of the stabilizer. The presented method provides a simplified and orderly surgical field without complicated devices and should help achieve a uniform and steady coronary anastomosis.

[Surgical treatment for aortic root dilatation and aortic regurgitation after arterial switch operation].

Aortic regurgitation (AR) and, aortic root dilatation are serious problems occurring after arterial switch operation (ASO). We have experienced a case of 12-year-old boy with aortic root dilatation and AR after ASO. Consistent progression of AR and aortic root dilatation were observed during the follow-up period. The latest Z-score of Valsalva sinus was about 9. We performed the Bentall procedure instead of aortic valve-sparing operation because of the degenerative aortic valve and the excessive aortic root dilatation. The postoperative course was uneventful. Histopathological findings of the aortic wall showed degeneration of elastic fibers on both sides of the anastmosis.

[Surgical repair of traumatic rupture of the discending aorta through 'L'-thoracotomy].

Acute blunt aortic rupture occurs frequently at the aortic isthmus and emergency operation is usually required. A 33-year-old man was suffered with blunt traumatic thoracic aortic injury caused by traffic accident and emergency operation was performed due to hemodynamic instability. The patient was operated through 'L'-thoracotomy (upper part sternotomy and antero-lateral thoracotomy). Cardiopulmonary bypass was initiated with right femoral vein drainage and right femoral arterial return and converted to standard cardiopulmonary bypass with the ascending aorta return and right atrium appendage drainage when the discending aorta was re-ruptured. It stabilized the circulation of upper body, especially brain. The post-operative course was uneventful. The 'L'-thoracotomy can provide good operative exposure for the aortic isthmus and stabilize the circulation of the brain with standard cardiopulmonary bypass and clamping the descending aorta.

Evaluation of circadian phenotypes utilizing fibroblasts from patients with circadian rhythm sleep disorders.

We evaluated the circadian phenotypes of patients with delayed sleep-wake phase disorder (DSWPD) and non-24-hour sleep-wake rhythm disorder (N24SWD), two different circadian rhythm sleep disorders (CRSDs) by measuring clock gene expression rhythms in fibroblast cells derived from individual patients. Bmal1-luciferase (Bmal1-luc) expression rhythms were measured in the primary fibroblast cells derived from skin biopsy samples of patients with DSWPD and N24SWD, as well as control subjects. The period length of the Bmal1-luc rhythm (in vitro period) was distributed normally and was 22.80±0.47 (mean±s.d.) h in control-derived fibroblasts. The in vitro periods in DSWPD-derived fibroblasts and N24SWD-derived fibroblasts were 22.67±0.67 h and 23.18±0.70 h, respectively. The N24SWD group showed a significantly longer in vitro period than did the control or DSWPD group. Furthermore, in vitro period was associated with response to chronotherapy in the N24SWD group. Longer in vitro periods were observed in the non-responders (mean±s.d.: 23.59±0.89 h) compared with the responders (mean±s.d.: 22.97±0.47 h) in the N24SWD group. Our results indicate that prolonged circadian periods contribute to the onset and poor treatment outcome of N24SWD. In vitro rhythm assays could be useful for predicting circadian phenotypes and clinical prognosis in patients with CRSDs.

[Aortic valve replacement for the small aortic annulus].

Aortic valve surgery for the small aortic annulus is still challenging for surgeons. Recently, the new types of high performance prosthesis have been developed and the chance of an aortic root enlargement (ARE) is decreasing. In this study, we propose the ideal strategy of the aortic surgery for the small aortic annulus. We analyzed the clinical records of 158 patients who underwent aortic valve replacement from August 1999 to October 2005 in our institution. The small aortic annulus was observed in 38 patients (24%). Fourteen patients of this group underwent ARE. Patient-prosthesis mismatch (PPM) was less frequently observed in patients with ARE compared to those without ARE. The additional time required for ARE was not considerable, and neither ischemic time nor cardiopulmonary bypass time was significantly prolonged by ARE. In conclusion, we have to select a prosthesis with sufficient orifice area to avoid PPM, otherwise we should choose an option of ARE. For this consideration, we definitely need the chart that demonstrates the relationship between the nominal size of various types of prostheses and the size of a patient's annulus that those prostheses actually fit.

Beta-enolase in blood plasma during open heart surgery.

The enolase isoenzyme composed of the beta subunit (alpha beta or beta beta enolase) is distributed predominantly in the heart and skeletal muscles. To see whether it may be a useful indicator of myocardial damage, concentrations of beta-enolase (the beta subunit of enolase) were determined in serial blood samples taken from 18 patients who underwent mitral valve replacement, using a highly sensitive enzyme immunoassay method. They were compared with those of creatine kinase MB isoenzyme (CK-MB) in the same samples. The mean arterial beta-enolase concentration was 6.60 (SD 3.84) ng.ml-1 at the beginning of anaesthesia. It increased rapidly after reperfusion and rose to greater than 100 ng.ml-1 at 2 h post-reperfusion. It remained high until 12 h post-reperfusion, and then decreased slowly. The plasma beta-enolase concentrations were significantly higher in coronary sinus samples than in arterial samples in the early phase after reperfusion. Since plasma carbonic anhydrase III, which is known to be localised only in the skeletal muscle, did not increase during the surgery, we suggest that the major portion of the elevated plasma beta-enolase was derived from heart muscle. Plasma concentrations of beta-enolase increased as quickly as those of CK-MB after reperfusion. The CK-MB concentrations had returned to normal at 2 d but the beta-enolase concentrations remained elevated up to 7 d. These results show that the determination of beta-enolase in plasma may be useful for estimating myocardial damage during open heart surgery.

A heat shock-related protein, p20, plays an inhibitory role in platelet activation.

Some low molecular mass heat shock proteins (HSPs) appear to act as molecular chaperones, but their exact physiological roles have not been fully elucidated. We reported previously that a 20-kDa protein (p20), which is classified as a low molecular mass HSP, is present at high levels in skeletal and smooth muscles. In the present study, we investigated a physiological role of p20 on platelet function in vitro and ex vivo. p20 inhibited platelet aggregation using human platelets dose-dependently induced by botrocetin. On the other hand, HSP27, the other type of low molecular mass HSP, did not affect platelet aggregation. When p20 (300 microg/kg) was injected intravenously as a bolus in hamsters, platelet aggregation ex vivo induced by botrocetin was also significantly inhibited. In order to further investigate the inhibitory effect by p20 on platelet activation, we performed platelet aggregation induced by thrombin or ADP using human platelets. p20 markedly prevented platelet aggregation induced by thrombin, but not ADP. These findings suggest that p20 can act intercellularly to regulate platelet functions. Our results may provide the basis for a novel defensive system to thrombus formation.

Regional cerebral tissue blood flow measured by the colored microsphere method during retrograde cerebral perfusion.

Brain tissue blood flow was measured precisely by the colored microsphere method during retrograde cerebral perfusion in 10 normothermic mongrel dogs. The average tissue blood flow rates to the cerebral cortex, cerebral medulla, brain stem, cerebellum, and spinal cord during retrograde cerebral perfusion at 25 mm Hg of external jugular venous pressure were 10.5 +/- 10.3, 4.2 +/- 4.6, 11.1 +/- 9.8, 12.3 +/- 8.6, and 9.1 +/- 5.8 ml/min per 100 gm, respectively. The brain was perfused wholly by retrograde cerebral perfusion without lateralization. Total cerebral blood flow was calculated as the sum total rates of blood flow to each area. Total cerebral blood flow during retrograde cerebral perfusion at 25 mm Hg was 7.8 +/- 4.4 ml/min, which represented 3.5% +/- 1.9% of whole body blood flow and one third of the total cerebral blood flow (28.0 +/- 4.2 ml/min) during cardiopulmonary bypass at a flow rate of 1000 ml/min. Oxygen consumption and carbon dioxide elimination by the total cerebrum during retrograde cerebral perfusion at 25 mm Hg were 0.54 +/- 0.23 ml/min and 34 +/- 15 mumol/min, respectively, or 8.6% +/- 3.6% and 7.0% +/- 3.1% of the corresponding whole body value and represented about one third of that measured during cardiopulmonary bypass (1.21 +/- 0.39 ml/min and 96 +/- 15 mumol/min). Total cerebral blood flow, total cerebral oxygen consumption, and carbon dioxide elimination increased as the external jugular venous pressure increased from 15 to 25 mm Hg; however, no further increase occurred once the external jugular venous pressure exceeded 25 mm Hg.

Balloon electric shock ablation. A surgical technique for treatment of ventricular tachycardia: influence of endocardial scar on depth of ablation achieved.

Balloon electric shock ablation is a surgical technique that has been used for treatment of ventricular tachycardia. However, little is known about the energy requirements and precise electrode array best suited to achieve effective ablation of the target area while limiting injury to the surrounding myocardium. This study was designed to determine the effects of endocardial scar (often present at the "site of origin" of clinical ventricular tachycardia) on depth of ablation resulting from balloon electric shock ablation. A chronic canine model of endocardial scar (3.9 +/- 0.6 mm thick) was developed with the use of balloon electric shock ablation techniques. We compared depth of ablation achieved with balloon electric shock ablation with low-energy shocks (22 J per bead) in normal dogs versus those with chronic endocardial scar. No difference was found in depth of ablation in normal dogs and in the scar model (7.2 +/- 1.2 mm versus 6.2 +/- 1.0 mm). Depth of injury expressed as a percentage of wall thickness was not different in the two groups (61% +/- 11% versus 57% +/- 3%). We conclude that the presence of endocardial scar does not influence depth of injury resulting from balloon electric shock ablation. This data provides guidance for clinical application of the technique as a "closed heart" surgical approach for control of ventricular tachycardia. The data are also discussed in relation to energy levels currently used for direct current catheter ablation in patients with ventricular tachycardia.

Comparative experimental study between retrograde cerebral perfusion and circulatory arrest.

To evaluate the efficacy of retrograde cerebral perfusion in protecting the brain, we comparatively studied retrograde cerebral perfusion and total circulatory arrest in 18 hypothermic (20 degrees C) mongrel dogs (retrograde cerebral perfusion, n = 10; total circulatory arrest, n = 8). Retrograde cerebral perfusion was performed, maintaining an external jugular venous pressure of 25 mm Hg for 60 minutes. Retrograde cerebral perfusion provided half the cerebral blood flow and a third of the oxygen that was supplied during hypothermic cardiopulmonary bypass, which had a flow rate of 100 ml/min per kilogram. Oxygen consumption and carbon dioxide exudation did not increase on resuming cardiopulmonary bypass after retrograde cerebral perfusion, whereas they increased after total circulatory arrest (oxygen consumption 10.7 +/- 5.3 versus 19.1 +/- 8.6 ml/min, p < 0.05; carbon dioxide exudation, 0.92 +/- 0.54 versus 1.64 +/- 0.78 mmol/min, p < 0.05). Therefore, oxygen debt during retrograde cerebral perfusion was smaller than during total circulatory arrest. Retrograde cerebral perfusion also cooled the brain better than did total circulatory arrest (20.4 degrees +/- 1.5 degrees C versus 22.7 degrees +/- 0.7 degrees C, p < 0.01). Cerebral tissue oxygen tension decreased slightly (27.5 +/- 7.7 versus 12.3 +/- 3.0 mm Hg, p < 0.01), and cerebral tissue carbon dioxide tension increased slowly during retrograde cerebral perfusion (95 +/- 34 versus 147 +/- 44 mm Hg, p < 0.05). These changes were smaller than those seen in total circulatory arrest. Tissue concentrations of adenosine triphosphate in the brain remained relatively high during retrograde cerebral perfusion but decreased rapidly during total circulatory arrest (0.49 +/- 0.16 versus 0.21 +/- 0.05 mmol/gm, p < 0.01, just before resuming cardiopulmonary bypass). Retrograde cerebral perfusion cannot maintain aerobic metabolism but may reduce ischemic damage of the brain and may safely extend the cerebral circulation interruption time.

Surgical treatment of ventricular tachycardia by balloon electric shock ablation. Potential effects on the mitral valve apparatus.

We have previously described a new surgical technique for control of arrhythmogenic foci in patients with recurrent ventricular tachycardia that we call balloon electric shock ablation. With this method sequential shocks are delivered to a grid of electrodes on a balloon that can be introduced across the mitral valve into the intact ventricle. A series of experiments was undertaken to investigate possible deleterious effects of balloon electric shock ablation when shocks are delivered directly to the mitral valve apparatus. In six animals shocks totaling 1200 joules were given through a closely spaced electrode grid applied to the area of the mitral valve. Nine to 12 weeks later, left ventricular and mitral valve function were assessed. Balloon electric shock ablation in the basilar portion of the ventricle was associated with decreased myocardial performance, as evidenced by ejection phase indices. In five of six animals balloon ablation led to minor thickening of the valve leaflets and chordal attachments plus necrosis of adjacent myocardium, including papillary muscles. In these animals there was no significant dysfunction of the valve observed. In the remaining animal, however, ablation was centered on the posterior papillary muscle and resulted not only in necrosis of the base of the papillary muscle but also in full-thickness scarring and thinning of the adjacent left ventricular wall. In this dog, mitral regurgitation was seen on long-term follow-up. We conclude that when balloon electric shock ablation is used to destroy a localized area of myocardium in the basilar portion of the intact ventricle, the procedure results in decreased myocardial performance. When shocks were directly applied to the mitral valve apparatus in five of six animals, ablation did not result in significant negative effects on the structure and function of the valve. In the sixth dog, however, shock delivery resulted in transmural necrosis and thinning at the site of papillary muscle insertion and was associated with severe mitral regurgitation with volume loading. Therefore caution should be used when considering clinical application of this technique if the area to be ablated is in the basal portion of the heart.

Body temperature influences regional tissue blood flow during retrograde cerebral perfusion.

OBJECTIVE: This study compared the cerebral microcirculation during retrograde cerebral perfusion with that during antegrade cardiopulmonary bypass under normothermic and hypothermic conditions. METHODS: Brain tissue blood flow was measured with the hydrogen-clearance and colored microsphere (15 and 50 microns) methods during antegrade cardiopulmonary bypass and retrograde cerebral perfusion. Measurements were performed during normothermia (37 degrees C), moderate hypothermia (28 degrees C) and deep hypothermia (20 degrees C) in groups of mongrel dogs (n = 8). RESULTS: During antegrade cardiopulmonary bypass, the microsphere method showed a significant decrease in cerebral blood flow as body temperature decreased (40.1 +/- 20.8 ml/min/100 gm at 37 degrees C, 16.2 +/- 18.0 ml/min/100 gm at 20 degrees C with 50 microns microspheres) At 20 degrees C, the cerebral blood flow measured with the 15 microns microspheres was significantly lower than that assessed with the hydrogen-clearance method (11.3 +/- 7.0 vs 24.8 +/- 7.0 ml/min/100 gm). During retrograde cerebral perfusion, the microsphere method also showed a significant decrease in cerebral blood flow with cooling. At 37 degrees C, the cerebral blood flow measured with the 15 microns microspheres (0.8 +/- 0.7 ml/min/100 gm) was significantly lower than that assessed with the hydrogen-clearance method (10.1 +/- 3.5 ml/min/100 gm). At both 28 degrees and 20 degrees C, the hydrogen-clearance method showed significantly higher cerebral blood flow (10.1 +/- 5.8 and 8.2 +/- 3.7 ml/min/100 gm) than did the 50 microns microspheres (1.8 +/- 0.6 and 1.0 +/- 0.8 ml/min/100 gm) and 15 microns microspheres (0.23 +/- 0.14 and 0.18 +/- 0.15 ml/min/100 gm). CONCLUSION: (1) Cerebral blood flow that shunts to capillaries is increased during antegrade cardiopulmonary bypass under deep hypothermia. (2) During retrograde perfusion, the majority of the blood flow shunts away from brain capillaries, even under normothermic conditions, and blood flow through large venoarterial shunts increases as body temperature decreases. Although the cerebral microcirculation during retrograde perfusion is decreased, retrograde perfusion provides some degree of oxygenation to the body.

Transatrial balloon technique for activation mapping during operations for recurrent ventricular tachycardia.

Results of operations for recurrent ventricular tachycardia have improved since methods of mapping that allow a directed approach to the problem have been developed. With standard operative techniques (ventriculotomy and introduction of a hand-held probe or multiple electrode array), it has not always been possible to obtain satisfactory endocardial activation maps during the tachycardia. We have recently developed a new transatrial balloon approach that has greatly facilitated intraoperative mapping. This paper describes our total experience with the new approach and draws attention to details of the cardiopulmonary bypass technique and the surgical approach needed for safe balloon insertion across the mitral valve. We describe how correlation between position of target electrodes on the balloon and the internal geometry of the heart is achieved and discuss the choice and application of appropriate ablation techniques. In our series of 37 consecutive patients, 35% had a grade IV ventricle (ejection fraction less than 20%), 32% had a previous posterior infarct, 51% did not have a resectable aneurysm, and 54% had been receiving amiodarone within 1 month of the operation. These factors have been associated with poor operative results in other series. With the transatrial balloon technique, we were able to induce and map ventricular tachycardia in 100% of patients (average 2.6 +/- 1.3 morphologies per patient). Using a variety of ablation techniques (endocardial excision, cryoablation, or balloon electric shock ablation), we have achieved surgical control of the arrhythmias in 84% of patients with an operative mortality rate of 14%. We recommend transatrial balloon mapping as the procedure of choice for intraoperative identification of arrhythmogenic foci in patients with recurrent ventricular tachycardia.

Determination of optimum retrograde cerebral perfusion conditions.

Retrograde cerebral perfusion through a superior vena caval cannula is a new technique used to protect the brain during operations on the aortic arch. We measured cerebral tissue blood flow, oxygen consumption, and cerebrospinal fluid pressure under various perfusion conditions in hypothermic (20 degrees C) mongrel dogs (n = 18, 12.8 +/- 0.6 kg) to determine the optimum conditions for retrograde cerebral perfusion. Retrograde cerebral perfusion was performed by infusion via the superior vena caval cannula and drainage via the ascending aortic cannula while the inferior vena cava and azygos vein were clamped. Retrograde cerebral perfusion was performed as the external jugular venous pressure was changed from 15 to 35 mm Hg in increments of 5 mm Hg. Cerebral tissue blood flow was measured by the hydrogen clearance method. Hypothermic retrograde cerebral perfusion with an external jugular venous pressure of 25 mm Hg provided about half the cerebral tissue blood flow of hypothermic (20 degrees C) cardiopulmonary bypass with a flow rate of 1000 ml/min (13.7 +/- 7.9 versus 32.7 +/- 8.5 ml/min per 100 gm). It decreased significantly as the external jugular venous pressure was decreased from 25 to 15 mm Hg but did not increase significantly as the external jugular venous pressure was increased from 25 to 35 mm Hg. Whole-body oxygen consumption during hypothermic retrograde cerebral perfusion with an external jugular venous pressure of 25 mm Hg was one quarter of that during hypothermic cardiopulmonary bypass (3.4 +/- 0.7 versus 12.7 +/- 5.6 ml/min) and varied in proportion to external jugular venous pressure. The cerebrospinal fluid pressure was a little lower than the external jugular venous pressure (19.2 +/- 4.5 mm Hg versus 24.8 +/- 2.4 mm Hg) but also varied with the external jugular venous pressure. The cerebrospinal fluid pressure remained lower than 25 mm Hg so long as the external jugular venous pressure remained lower than 25 mm Hg. High external jugular venous pressure was associated with high intracranial pressure, which restricts cerebral tissue blood flow and may cause brain edema. We believe that a venous pressure of 25 mm Hg is the optimum condition for retrograde cerebral perfusion.

Detection of interstitial Ga in GaN

We report the direct detection of interstitial Ga by optical detection of electron paramagnetic resonance (ODEPR) in the photoluminescence of n-type GaN after irradiation in situ at 4.2 K with 2.5 MeV electrons. It is stable upon annealing until room temperature, where it becomes mobile and trapped to form a new defect which is observed to emerge as the interstitial disappears. The time constant of the process at room temperature is approximately 200 min. The emergence of another ODEPR center beginning at approximately 135 K suggests even easier migration of one of the other intrinsic defects in the GaN lattice.