Hypothermic preservation of rat hearts using antifreeze glycoprotein.

Antifreeze proteins are an effective additive for low-temperature preservation of solid organs. Here, we compared static hypothermic preservation with and without antifreeze glycoprotein (AFGP), followed by nonfreezing cryopreservation of rat hearts. The heart was surgically extracted and immersed in one of the cardioplegia solutions after cardiac arrest. Control rat hearts (n=6) were immersed in University of Wisconsin (UW) solution whereas AFGP-treated hearts (AFGP group) (n=6) were immersed in UW solution containing 500 ?g/ml AFGP. After static hypothermic preservation, a Langendorff apparatus was used to reperfuse the coronary arteries with oxygenated Krebs-Henseleit solution. After 30, 60, 90, and 120 min, the heart rate (HR), coronary flow (CF), cardiac contractile force (max dP/dt), and cardiac diastolic force (min dP/dt) were measured. Tissue water content (TWC) and tissue adenosine triphosphate (ATP) levels in the reperfused preserved hearts were also assessed. All the parameters were compared between the control and AFGP groups. Compared with the control group, the AFGP group had significantly (p<0.05) higher values of the following parameters: HR at 60, 90, and 120 min; CF at all four time points; max dP/dt at 90 min; min dP/dt at 90 and 120 min; and tissue ATP levels at 120 min. TWC did not differ significantly between the groups. The higher HR, CF, max dP/dt, min dP/dt, and tissue ATP levels in the AFGP compared with those in control hearts suggested that AFGP conferred superior hemodynamic and metabolic functions. Thus, AFGP might be a useful additive for the static/nonfreezing hypothermic preservation of hearts.

Persistent spontaneous pneumothorax for four years: a case report.

Pneumothorax, defined as the presence of air in the pleural space, is usually classified as spontaneous or traumatic; it is unusual for pneumothorax to be categorized as being acute or chronic. Even if conservative treatment is chosen, the pneumothorax is cured when air in the pleural space dissolves into the venous blood. A 50-years-old Japanese man with no prior medical history was referred to our department with a right pneumothorax and two rightsided pulmonary nodules on chest X-ray and CT. The chest radiographs of past mass screening which was taken four years ago showed right pneumothorax and right-sided pulmonary nodules. From then, all chest radiograph and chest computed tomography showed right pneumothorax and pulmonary nodules. But he underwent no medical interventions. We designed to perform an operation for a treatment of right pneumothorax and the diagnosis of pulmonary tumors. We underwent right upper lobectomy and pleural decortication under video assisted thoracic surgery. We obtained pathological diagnosis of inflammatory pseudotumor and surrounding atelectasis. He was cured from pneumothorax and pulmonary tumors. A unique case of spontaneous pneumothorax presenting with a pleural air space that was confirmed by chest radiographs and computed tomography examinations over a 4-year period is reported.

Selective blockade of apoptosis by in vivo electroporation-mediated gene transfer combined with portal infusion of plasmid DNA attenuates liver cirrhosis.

AIM: The purpose of the present study was to determine whether in vivo electroporation could achieve selective blockade of apoptosis in a rat liver cirrhosis model. METHODS: A dimethylnitrosamine (DMN)-induced rat liver cirrhosis model was used. In vivo electroporation was performed after portal vein infusion of plasmid DNA. pFas-Fc plasmid DNA was used to block the apoptotic pathway. pUC/HGF and pCAGGS/EGFP were used as positive and negative controls, respectively. Liver collagen content was evaluated by hydroxyproline assay two weeks after gene transfer. Terminal deoxynucleotidyltransferase dUTP nick end-labeling was simultaneously performed in the liver to evaluate suppression of apoptosis. Survival analysis was performed using 10 rats that received the sFas gene, 10 that received the HGF gene, and 13 that received the GFP gene. RESULTS: The apoptotic cell index in the DMN-injected liver was significantly lower in rats that received the sFas gene compared with the negative control. The collagen content of the DMN-injected liver was also lower in rats that received the sFas gene compared with the negative control. There was no significant difference in the apoptotic cell index and collagen content of rats that received the sFas and HGF genes. Ten weeks after the initiation of DMN treatment, the survival rates with the sFas, HGF, and GFP genes were 56%, 100%, and 0, respectively. CONCLUSION: Selective blockade of apoptosis by in vivo electroporation-mediated gene transfer improved the apoptotic cell index, hydroxyproline content, and survival rate. Soluble Fas gene therapy using in vivo electroporation can be a safe and efficient therapy for liver cirrhosis in rats.

[Reconstruction of the left common carotid artery for dissective occlusion detected by intra-operative ultrasonography].

We describe a case of Stanford type A acute aortic dissection. Replacement of the ascending aorta and aortic arch was performed using an "arch 1st technique". Following the completion of replacement, hypotension of the left superficial temporal artery pressure was detected. Ultrasonography revealed dissection of the left common carotid artery (LCCA) and compressive occlusion of the true lumen. Reconstruction of the LCCA was performed in the neck. The patient did well after the operation without any neurological abnormalities.

Two-stage portal vein ligation facilitates liver regeneration in rats.

BACKGROUND/AIMS: Recent reports have demonstrated that some patients are unable to undergo scheduled liver resection after preoperative portal vein embolization due to insufficient hypertrophy of the future remnant liver. The present study examined whether two-stage portal vein ligation (PVL) increases hypertrophy of the future remnant liver compared to conventional PVL in rats. METHODS: Rats were divided into 3 groups: group A, ligation of left primary branch; group B, ligation of right and left primary branches; group C, ligation of the left primary branch, followed by 2-stage PVL 7 days postoperatively. To evaluate liver regeneration, the proliferating cell nuclear antigen labeling index (LI), mitotic index (MI) in the caudate lobe and weight ratio of caudate lobe to body weight were measured. RESULTS: The weight ratio of caudate lobe to body weight was significantly higher in group C than in groups A or B 14 days postoperatively. In groups A and B, LI and MI in the caudate lobe peaked 2 days postoperatively, then decreased to preoperative levels by 7-8 days postoperatively, but remained significantly elevated until 10-14 days postoperatively in group C. CONCLUSION: Two-stage PVL increases hypertrophy of the future remnant liver compared to conventional PVL in rats.

The intracellular accumulation of phagocytic and epithelial cells and the inhibitory effect on Chlamydophila (Chlamydia) pneumoniae of telithromycin and comparator antimicrobials.

The intracellular accumulation of telithromycin was measured and compared with ciprofloxacin, clarithromycin, minocycline and erythromycin. The activities of telithromycin, clarithromycin and minocycline against Chlamydophila (Chlamydia) pneumoniae were compared in an intracellular killing assay. Maximal telithromycin accumulation (mean intracellular/extracellular concentration ratio) ranged from 6.7 (A549 lung epithelial cells) to 11.8 (THP-1 monocytic cells). This ratio was similar to that of clarithromycin, but less than for minocycline. Minimum inhibitory and minimum lethal telithromycin concentrations against six clinical strains of C. pneumoniae were <0.015-0.03 mg/L and 0.03-0.06 mg/L, respectively, which were lower than those found for minocycline. These results show that telithromycin accumulates rapidly into epithelial cells, similar to previously reported results for phagocytic cells. Intracellular accumulation of telithromycin was lower than that observed for minocycline, but telithromycin demonstrated substantially more potent antichlamydial activity.

[Anaphylactic shock related to aprotinin induced by anti-aprotinin immunoglobulin G antibody alone; report of a case].

Anaphylactic shock related to aprotinin has been reported to be induced exclusively in the presence of IgE antibody. And the possibility of anaphylactic shock induced by anti-aprotinin IgG antibody alone was controversial. In this paper, we describe the first case of anaphylactic shock induced by aprotinin-specific IgG antibody alone. A 55-year-old man underwent surgical repair of the descending aorta with the use of aprotinin at 2 months after first aprotinin usage. Immediately after initiation of cardiopulmonary bypass with the continuous infusion of aprotinin, clinical symptoms of anaphylactic reaction were found. Postoperative drug lymphocyte stimulation test for aprotinin and aprotinin-specific IgE antibody were negative, but aprotinin-specific IgG antibody was 163 mg/l and positive.

p21CDKN1A allows the repair of replication-mediated DNA double-strand breaks induced by topoisomerase I and is inactivated by the checkpoint kinase inhibitor 7-hydroxystaurosporine.

This study provides evidence for the importance of p21(CDKN1A) for the repair of replication-mediated DNA double-strand breaks (DSBs) induced by topoisomerase I. We report that defects of p21(CDKN1A) and p53 enhance camptothecin-induced histone H2AX phosphorylation (gammaH2AX), a marker for DNA DSBs. In human colon carcinoma HCT116 cells with wild-type (wt) p53, gammaH2AX reverses after camptothecin removal. By contrast, gammaH2AX increases after camptothecin removal in HCT116 cells deficient for p53 (p53-/-) or p21(CDKN1A) (p21-/-) as the cells reach the late-S and G2 phases. Since p21-/- cells exhibit similar S-phase arrest as wt cells in response to camptothecin and aphidicolin does not abrogate the enhanced gammaH2AX formation in p21-/- cells, we conclude that enhanced gammaH2AX formation in p21-/- cells is not due to re-replication. The cell cycle checkpoint abrogator and Chk1/Chk2 inhibitor 7-hydroxystaurosporine (UCN-01) also increases camptothecin-induced gammaH2AX formation and inhibits camptothecin-induced p21(CDKN1A) upregulation in HCT116 wt cells. TUNEL (terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling) assays demonstrate that gammaH2AX formation in late S and G2 cells following CPT treatment corresponds to DNA breaks. However, these breaks are not related to apoptotic DNA fragmentation. We propose that p21(CDKN1A) prevents the collapse of replication forks damaged by stabilized topoisomerase I cleavage complexes.

Comparison of NSP4 protein between group A and B human rotaviruses: detection of novel diarrhea-causing sequences in group B NSP4.

The human group B rotavirus is a causative agent of severe adult diarrhea. In this study, we analyzed the NSP4 structure of a group B rotavirus strain, CAL-1, and determined whether enterotoxin activity was present in CAL-1 NSP4. CAL-1 NSP4 was comprised of 219 amino acids which was longer than group A and C rotavirus NSP4, and the primary structures of their sequences differed considerably. However, CAL-1 NSP4 had an enterotoxin-like sequence (residues 106-127) that was only 27% identical to the enterotoxin region of NSP4 of KUN (a group A rotavirus strain) at residues 114-135. Interestingly, both of the synthetic peptides, one (residues 99-128) containing the enterotoxin-like sequence and the other (residues 191-219) containing 29 C-terminal amino acids of CAL-1 NSP4, induced diarrhea in 5.5-day-old mice, but not in 17.5-day-old mice, when administered parenterally. Thus, rotavirus "enterotoxin" sequences could be considerably divergent.

Evaluation of a fully automated system (RAISUS) for rapid identification and antimicrobial susceptibility testing of Staphylococci.

RAISUS is a system for rapid bacterial identification and antimicrobial susceptibility testing. RAISUS and VITEK showed 97.8% and 75.9% agreement in identification of 45 Staphylococcus aureus strains and 58 coagulase-negative staphylococci (CoNS), respectively, and RAISUS and CLSI (formerly NCCLS) methods showed 87.2% and 87.9% agreement in the MICs for S. aureus and CoNS, respectively. RAISUS provided these data within 3.75 h, suggesting its utility for clinical bacteriological laboratories.

[Arch first technique using handmade branched graft].

We created a method using handmade branched graft to do the aortic arch surgery easier and safer. We made the branched graft using 12 and 8 mm vascular graft. A 77-year-old man with Stanford type A aortic dissection was operated with this method under deep hypothermia. After aortic root manipulation, perfusion of the aortic arch was stopped and selective cerebral perfusion was established. Left subclavian artery (LSCA) was anastomosed to one of the branches. The perfusion of the LSCA was re-started via one of its branches. Respectively, left common carotid artery and brachiocephalic artery reconstruction and reperfusion were performed in a same fashion. After distal anastomosis, anastomosis between the branched graft and main graft was performed consecutively. Postoperative course was uneventful and there was no complication. The treatment of our branched graft was easier than that of ready-made 4-branched graft. We could perform the operation under clear view for its movability with minimal cerebral ischemic time.

Average P/F ratio during donor management for finding out the potential donor of lung procurement.

We investigated the potential donors from cases without lung procurement in spite of considering procurement as the donor. We reviewed 207 cases considered for lung procurement. Donors were divided into 2 groups according to whether or not their lungs were harvested: 50 did and 157 did not. We investigated their age, gender, donor management periods, blood pressure, heart rate, P/F ratio, PCO2, HCO3, positive end-expiratory pressure (PEEP), respiratory rate, abnormal findings in the chest X-ray, blood chemistries, sputum culture, and intravenous administration of steroid. Univariable or multivariable logistic regression analysis was used to predict lung harvesting by various factors. Univariable logistic regression analysis revealed maximum heart rate (HR) and respiratory rate (RR), maximum, minimum, and average P/F ratio, and abnormal findings in the chest X-ray, to be predictors to perform transplant. The maximum HR and RR of the non-harvest group are higher than those of the harvest group. Multivariable logistic regression analysis revealed average P/F ratio only to be a predictor to transplant. We divided the non-harvest group into 2 groups according to whether or not their P/F ratio was >300: 55 did (P/F 300 group) and 102 did not (P/F 299 group). Between P/F 299 group and the harvest group, univariable logistic regression analysis revealed maximum HR, maximum and minimum RR, maximum PEEP, maximum, minimum, and average P/F ratio, and abnormal findings in the chest X-ray to be predictors to transplant. Maximum HR, PEEP, and maximum and minimum RR of the P/F 299 group are higher than those of the harvest group. Maximum, minimum, and average P/F ratio of the P/F 299 group are lower than those of the harvest group. The average PCO2 of P/F 299 group is higher than that of the harvest group. The rate of abnormal findings in the chest X-ray of the P/F 299 group is higher than that of the harvest group. Multivariable logistic regression analysis revealed average P/F ratio only to be a predictor to transplant. There were no predictors between the P/F 300 group and the harvest group in both univariable and multivariable logistic regression analyses. Furthermore, there were no different factors between the P/F 300 group and the harvest group. Our study showed that 35.5% of the non-harvest group had a P/F ratio of >300 and had no difference compared with the harvest group. Our data suggest that potential donors existed in the non-harvest group and increased the number of lung procurement to 27.3% from 13.0% of all donors.

[Clinical experience with aortic connector system].

From April to December 2002, 40 patients underwent coronary artery bypass grafting (CABG) using the St. Jude Medical (Minneapolis) Symmetry bypass system (aortic connector system: ACS). 59 proximal anastomoses (51 saphenous vein grafts, 8 radial artery grafts) were performed with the ACS. One saphenous vein graft occluded during operation. Postoperative evaluation of the anastomotic patency was carried out by angiography in 45 grafts. Five of the saphenous vein grafts were occluded (5/38). One patient who was shock state before operation presented with postoperative unconsciousness. Another patient died at 8th postoperative day caused by ventricular fibrillation. We conclude that the ACS produces a simple, quick way of performing the proximal anastomosis without the need for clamping the aorta, allows reducing risk of embolization by aortic manipulation. However, it is necessary to discuss sufficiently using the ACS, because the graft patency with the ACS is lower than with standard suturing technique.

[Surgery for acute aortic dissection concomitant with preceding axillofemoral bypass to prevent malperfusion of visceral organs and limb ischemia].

A 52-year-old man was admitted with sudden onset of epigastralgia. Abdominal X-ray showed dilated intestine and computed tomography (CT) revealed extended type A aortic dissection. Marked abdominal distention and weak pulse of right femoral artery were recognized so malperfusion of visceral organs due to narrowing true lumen compressed by thrombosed false lumen was suggested. In the operation, right axillo-right femoral bypass was established preceding to median sternotomy. This graft was used as an arterial perfusion site of cardiopulmonary bypass, and replacement of the ascending aorta was performed under hypothermic circulatory arrest and retrograde cerebral perfusion. Sign of malperfusion of visceral organs was showed for several days after the operation but it disappeared without further intervention. Axillofemoral inflow of cardiopulmonary bypass may be effective procedure in these cases.

Effects of vitamin supplementation on microcirculatory disturbance in hemodialysis patients without peripheral arterial disease.

AIMS: Dysfunctional endothelium caused by oxidative stress is thought to play a role in pathogenesis of a variety of conditions including atherosclerosis. We investigated whether a microcirculatory disturbance in hemodialysis (HD) patients was associated with increased oxidative stress and endothelial injury. PATIENTS AND METHODS: Transcutaneous oxygen tension (TcPO2) on the dorsum of the foot at rest was measured as a marker of microcirculation in 33 patients undergoing HD without clinical manifestations of peripheral arterial disease and 20 healthy controls. Furthermore, in order to examine whether TcPO2 was affected by antioxidants, oral supplementation with a combination of vitamin C (200 mg daily) and vitamin E (600 mg daily) was administered for 6 months to 8 patients with microcirculatory disturbance (TcPO2 values of 50 mmHg or less). Serum biochemical parameters including vitamins were also measured. RESULTS: Mean TcPO2 value was significantly lower in HD patients than in control subjects (47.9 +/- 13.5 mmHg versus 62.4 +/- 11.9 mmHg, p < 0.001). After vitamin supplementation, TcPO2 values remarkably increased (40.6 +/- 10.0 mmHg versus 57.4 +/- 6.5 mmHg, p < 0.005). Serum vitamin C and vitamin E levels increased significantly as well, while serum levels of thrombomodulin, a marker of endothelial injury, and thiobarbituric acid reactants, a marker of lipid peroxidation, were significantly decreased in comparison with those before supplementation. CONCLUSIONS: Our results suggest that the microcirculatory disturbance in HD patients seems to be associated with endothelial damage caused by oxidative stress. Combined supplementation with vitamin C and vitamin E may be of clinical benefit in improving the cutaneous microcirculation by reducing oxidative stress.

Multiple minimally invasive direct CABG for the complete revascularization: the figure L approach.

BACKGROUND: Coronary artery bypass grafting on the beating heart via median sternotomy causes significant hemodynamic compromise during displacement of the heart, especially in patients with low ejection fraction or those who have potential catastrophic sternal reentry injuries. We have developed an innovative minimally invasive surgical approach (figure L Approach) for the treatment of multi-vessel coronary artery disease; especially in cases with huge hearts (> CTR 50 %) including tracheostomy requiring COLD, and redo cases. The objective of this study was to assess the efficacy and safety of this alternative surgical incision. METHODS: From January 1998 to March 2001, 22 patients underwent complete revascularization using this figure L Approach. Left submammary anterior thoracotomy incisions are made with the medial part of the incision vertically extending downward to the upper abdomen. The costal margin was divided. The pleural and peritoneal spaces were then entered. A chest retractor was placed to elevate the chest wall, exposing the heart and stomach. Two to four arterial grafts including LIT A, RGEA, RA and IEA were harvested. Revascularization of the LAD, LCX and distal RCA were performed in 22 patients without cardiopulmonary bypass on complex performed arterial grafts using this approach. In four patients, triple and quadruple vessel grafting was performed. RESULTS: No early deaths or postoperative complications occurred. The mean coronary clamp time was 12.5 +/- 5.1 minutes for the LAD, 10.8 +/- 1.8 minutes for diagonal branch, 14.1 +/- 2.9 minutes for the LCX and 16.0 +/- 5.1 minutes RCA time. There were no late deaths or angina during the mean follow-up of 37.3 +/- 9.0 months. Postoperative coronary angiography demonstrated widely patent grafts in all but one patient. Hemodynamics did not change significantly during the distal anastomoses. No wound-healing problems were experienced. CONCLUSIONS: Multiple MIDCABG using the figure L Approach represents a novel way of approaching both the LAD, CX and distal RCA in patients with multi-vessel disease, especially in those with low ejection fraction, huge heart (> CTR 50 %), or hemodynamically unstable patients, without any hemodynamic deterioration or sternal related complication.

Behçet's disease accompanied by myelodysplastic syndrome with trisomy 8: two case reports and a review of 15 Japanese cases.

Abstract We describe two cases of Behçet's disease associated with myelodysplastic syndrome (MDS) with trisomy 8. Both cases developed ulceration in the cecum as a gastrointestinal complication of Behçet's disease, after a diagnosis of MDS. We summarized recent case reports of Behçet's disease associated with myelodysplastic syndrome, and studied the clinical manifestations. Most cases showed trisomy 8 as a chromosomal abnormality. Gastrointestinal involvement without eye lesions seems to be characteristic of Behçet's disease associated with MDS.

Serosal application of Ba(2+) induces oscillatory chloride secretion via activation of submucosal cholinergic neurones in guinea-pig distal colon.

The enteric nervous system regulates ion and fluid secretion in the mammalian intestine at both resting and stimulated conditions. To determine the type and activation mechanism of neurones involved, mucosa-submucosa sheets isolated from guinea-pig distal colon were studied in vitro in Ussing chambers. Serosal addition of 0.5-1 mM barium (Ba(2+)), a potassium (K(+)) channel inhibitor, caused oscillatory increases in short-circuit current (I(sc)). Mean values of the size and frequency of I(sc) were 369.1 microA cm(-2) and 2.3 min(-1). The oscillatory I(sc) induced by the low concentrations of Ba(2+) was blocked by either higher concentrations of Ba(2+) (2-5 mM) or other K(+) channel inhibitors, such as tetraethylammonium (TEA) (1 mM) and quinine (20 mM). The Ba(2+)-induced oscillatory I(sc) was also inhibited by tetrodotoxin (TTX) and atropine. In a nominally Ca(2+) free solution plus serosal addition of 0.1 mM ethylene glycol-bis (beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA), a Ca(2+) chelator, the oscillatory I(sc) slowed and diminished. Further, the Ba(2+)-induced oscillatory I(sc) was partially inhibited by apical addition of 100 microM 5'-nitro-2-(3-phenylpropylamino)benzoic-acid (NPPB), a Cl(-) channel inhibitor, and completely disappeared in a low Cl(-) solution (11 mM) on both sides. On the other hand, application of either cimetidine, a histamine H(2) receptor antagonist, or hexamethonium, a nicotinic antagonist, to the serosal side did not affect the Ba(2+)-induced oscillatory I(sc). In conclusion, the Ba(2+)-induced oscillatory I(sc) is the transepithelial Cl(-) current which is stimulated by activation of cholinergic neurones in submucosal plexus of guinea-pig distal colon.

An HLA-B27-positive patient diagnosed with ulcerative colitis 15 years after the onset of arthropathy.

Abstract A 28-year-old woman had persistent pain of both hip joints since the age of 13 years. X-ray analysis showed destructive changes in both hip joints and ossification of sacroiliitic joints. The patient had mild diarrhea and slight abdominal pain for 8 years. Blood-stained stool was not noticed. Barium enema showed changes consistent with the diagnosis of ulcerative colitis (UC). Inflammatory bowel syndrome should be considered in patients with persistent coxitis, even in the absence of severe abdominal symptoms.

Captopril renography in Bartter's syndrome.

We describe a 19-year-old man who developed muscle weakness due to hypokalemia. His blood pressure on admission was considered to be relatively high, and we performed captopril renography. His baseline renogram was normal, while the results of captopril renography showed delayed patterns similar to those of renovascular hypertension. However, his blood pressure level decreased and returned to the normal range without medication 2 weeks after admission. Our final diagnosis in this patient was Barter's syndrome. We speculate that the delayed pattern of captopril renography was due to an imbalance between the prostaglandin and the renin-angiotensin systems. The blockade of the renin-angiotensin system by captopril may cause dominant prostaglandins, produce systemic vasodilatory effects, and lead to hypoperfusion of the kidneys in patients with Bartter's syndrome.