Alterations of quantitative EEG and mini-mental state examination in interferon-alpha-treated hepatitis C.

We have recently observed a diffuse slowing of brain waves using serial quantitative electroencephalographic (qEEG) examinations in interferon (IFN)-alpha-treated chronic hepatitis C patients. However, it remains unclear how this alteration could be assessed. We evaluated the correlation between the qEEG changes and three tests of mental status, including the Mini-Mental State Examination (MMSE), in such patients. This is the first study to undertake a clinical evaluation of the adverse effects on brain function due to IFN. We undertook blind, prospective and serial qEEG examinations on 56 chronic hepatitis C patients at three independent hospitals. IFN-alpha was administered intramuscularly at a dose of 9 x 10(6) IU daily for the first 4 weeks and then 3 times/week for the next 20 weeks. Serial EEGs were obtained before, at 2 and 4 weeks of treatment, and after the IFN-alpha treatment. The absolute power values of each frequency band in each patient at different stages of treatment were recorded by qEEG. Each patient was assessed by the MMSE, Hamilton Rating Scale for Depression (HSD), and Hamilton Rating Scale for Anxiety (HSA). We statistically evaluated the correlations between the changes in power values and alterations of scores on the mental status tests during IFN-alpha treatment. The decreased scores observed on the MMSE ranged from 2 to 5 points at both 2 and 4 weeks of IFN-alpha treatment. There were no significant differences in age distribution for each decreased score on the MMSE. As the alteration in MMSE score during IFN treatment increased, the alteration in absolute power values for the slow waves during IFN treatment increased significantly, while that for the alpha 2 and fast waves during treatment decreased significantly. However, the changes in the HDS and HSA revealed no significant correlations. The alteration of the qEEG was reversible after the treatment. MMSE scores represent one screening method for the clinical assessment of IFN-alpha-induced alterations of brain function.

Genetic analysis of pancreatic duct hyperplasia in Otsuka Long-Evans Tokushima Fatty rats: possible association with a region on rat chromosome 14 that includes the disrupted cholecystokinin-A receptor gene.

An Otsuka Long-Evans Tokushima Fatty (OLETF) strain of rat spontaneously developed hyperglycemia, hyperinsulinemia, insulin resistance and mild obesity, which had been studied as animal model for type II diabetes mellitus (T2DM). Recently, we observed that this strain coincidentally developed atypical hyperplasia of the choledocho-pancreatic ductal epithelium with a complete incidence. In an effort to locate genes responsible for this hyperplasia, we prepared 288 backcross progeny from a mating between OLETF rats and BN rats (which do not develop hyperplasia), and performed a genome-wide scan using 207 polymorphic genetic markers. We observed a prominent association of hyperplasia with a region involving a marker locus D14Mit4 (P = 0.00020, Fisher's exact test) and Cckar (the cholecystokinin-A receptor gene; P = 0.00025, Fisher's exact test) which is known to be disrupted in an OLETF strain. Our findings indicated that epithelial hyperplasia of the choledocho-pancreatic duct is associated with a region on rat chromosome 14 around the Cckar gene in an additive fashion with another two susceptible loci, each on chromosome 9 and 7. This implied the possibility that Cckar deficiency could result in a predisposition towards pancreatic duct hyperplasia.

Effects of Dmo1 on obesity, dyslipidaemia and hyperglycaemia in the Otsuka Long Evans Tokushima Fatty strain.

Whole-genome scans have identified Dmo1 as a major quantitative trait locus (QTL) for obesity and dyslipidaemia in the Otsuka Long Evans Tokushima Fatty (OLETF) rat. We have produced congenic rats for the Dmo1 locus, using marker-assisted speed congenic protocols, enforced by selective removal of other QTL regions (QTL-marker-assisted counterselection), to efficiently transfer chromosomal segments from non-diabetic Fischer 344 (F344) rats into the OLETF background. In the third generation of congenic animals, we observed a substantial therapeutic effect of the Dmo1 locus on lipid metabolism, obesity control and plasma glucose homeostasis. We conclude that single-allele correction of an impaired genetic pathway can generate a substantial therapeutic effect, despite the complex polygenic nature of type II diabetic syndromes.

Single-allele correction of the Dmo1 locus in congenic animals substantially attenuates obesity, dyslipidaemia and diabetes phenotypes of the OLETF rat.

1. Whole-genome scans have identified Dmo1 as a major quantitative trait locus for dyslipidaemia and obesity in the Otsuka Long Evans Tokushima Fatty (OLETF) rat. 2. We have produced congenic rats for the Dmo1 locus through successive back-cross breeding with diabetic OLETF rats. Marker-assisted speed congenic protocols were applied to efficiently transfer chromosomal segments from non-diabetic Brown Norway (BN) rats into the OLETF background. 3. In the fourth generation of congenic animals, we observed a substantial therapeutic effect of the Dmo1 locus on lipid metabolism, obesity control and plasma glucose homeostasis. 4. We have concluded that Dmo1 primarily affects lipid homeostasis, obesity control and/or glucose homeostasis at fasting and is secondarily involved in glucose homeostasis after loading. 5. The results of the present study show that single-allele correction of a genetic defect of the Dmo1 locus can generate a substantial therapeutic effect, despite the complex polygenic nature of type II diabetic syndromes.

Quantitative trait loci for lipid metabolism in the study of OLETF x (OLETF x Fischer 344) backcross rats.

1. The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is a model of type II diabetes with accompanying dyslipidaemia and obesity. 2. To define chromosomal intervals associated with obesity (abdominal fat weight and plasma leptin levels), dyslipidaemia (plasma triglyceride, cholesterol and free fatty acids) and hyperglycaemia (plasma glucose levels), we have performed genome-wide quantitative traits loci (QTL) analyses of 115 male OLETF x (OLETF x Fischer 344) backcross animals at 16 weeks of age. 3. The Diabetes Mellitus OLETF type I (Dmo1) locus on rat chromosome 1 showed statistically significant involvement in elevations of plasma levels of triglycerides (P = 4.87 x 10(-6) at D1Rat90) and total cholesterol (P = 1.16 x 10(-5) at D1Rat306). 4. No other loci produced significant linkage to these observed phenotypes. 5. These analyses have confirmed the importance of Dmo1 in lipid homeostasis at younger ages as well as during overt diabetes, which appears later. Thus, alterations at the Dmo1 locus are a major risk factor for pathogenesis in the strain, a finding that agrees with physiological studies that indicate a role for dyslipidaemia in the type II diabetic syndrome of OLETF rats.

Characterization of newly developed SSLP markers for the rat.

We have isolated more than 12,000 clones containing microsatellite sequences, mainly consisting of (CA)n dinucleotide repeats, using genomic DNA from the BN strain of laboratory rat. Data trimming yielded 9636 non-redundant microsatellite sequences, and we designed oligonucleotide primer pairs to amplify 8189 of these. PCR amplification of genomic DNA from five different rat strains yielded clean amplification products for 7040 of these simple-sequence-length-polymorphism (SSLP) markers; 3019 markers had been mapped previously by radiation hybrid (RH) mapping methods (Nat Genet 22, 27-36, 1998). Here we report the characterization of these newly developed microsatellite markers as well as the release of previously unpublished microsatellite marker information. In addition, we have constructed a genome-wide linkage map of 515 markers, 204 of which are derived from our new collection, by genotyping 48 F2 progeny of (OLETFxBN)F2 crosses. This map spans 1830.9 cM, with an average spacing of 3.56 cM. Together with our ongoing project of preparing a whole-genome radiation hybrid map for the rat, this dense linkage map should provide a valuable resource for genetic studies in this model species.

Genetic dissection of "OLETF," a rat model for non-insulin-dependent diabetes mellitus: quantitative trait locus analysis of (OLETF x BN) x OLETF.

To identify genetic determinants relevant to non-insulin-dependent diabetes mellitus (NIDDM), we performed a genome-wide analysis for quantitative trait loci (QTLs) using 359 backcross progeny of the Otsuka Long-Evans Tokushima Fatty (OLETF) rat. The OLETF strain is a well-studied animal model of obese NIDDM, with features of hyperinsulinemia, hyperglycemia, insulin resistance, and abundant abdominal fat. Our extensive genomic scanning with 218 markers revealed nine significant QTLs, including a strong determinant of obesity on chromosome 1 (Dmo1: LOD = 13.99, for body weight). Two highly significant QTLs for glucose homeostasis were found, one on chromosome 1 (Dmo4 LOD = 7.16, for postprandial glucose level) and the other on chromosome X (Dmo11/Odb1: LOD = 7.81, for postprandial glucose level). These data are comparable to results of our previous studies of the OLETF rat.

A radiation hybrid map of the rat genome containing 5,255 markers.

A whole-genome radiation hybrid (RH) panel was used to construct a high-resolution map of the rat genome based on microsatellite and gene markers. These include 3,019 new microsatellite markers described here for the first time and 1,714 microsatellite markers with known genetic locations, allowing comparison and integration of maps from different sources. A robust RH framework map containing 1,030 positions ordered with odds of at least 1,000:1 has been defined as a tool for mapping these markers, and for future RH mapping in the rat. More than 500 genes which have been mapped in mouse and/or human were localized with respect to the rat RH framework, allowing the construction of detailed rat-mouse and rat-human comparative maps and illustrating the power of the RH approach for comparative mapping.

Genetic determinants of plasma triglyceride levels in (OLETF x BN) x OLETF backcross rats.

Altered lipid metabolism is closely associated with diabetes in humans, although predisposing genetic factors that affect hyperlipidemia have not yet been clarified. Our previously established OLETF strain is an obese rat model of type II diabetes, exhibiting hypertriglycemia as well as hyperinsulinemia, hyperglycemia, insulin resistance, and abundant abdominal fat. To identify genetic factors responsible for dyslipidemic phenotypes in OLETF rats, we performed a whole-genome scan using 293 male (OLETF x BN) x OLETF backcross rats. Our analysis identified two significant quantitative trait loci (QTLs), on rat chromosomes 1 and 8, that are related to fasting triglyceride levels. The chromosome 1 QTL colocalized with Dmo1 (diabetes mellitus, OLETF type 1), a locus previously shown to associate strongly with both fat levels and body weight. The other significant QTL localizes to the chromosome 8 marker D8Mit2, in a region where several apo-lipoprotein genes are clustered.

Genetic dissection of "OLETF", a rat model for non-insulin-dependent diabetes mellitus.

To elucidate the genetic factors underlying non-insulin-dependent diabetes mellitus (NIDDM), we performed genome-wide quantitative trait locus (QTL) analysis, using the Otsuka Long-Evans Tokushima Fatty (OLETF) rat. The OLETF rat is an excellent animal model of NIDDM because the features of the disease closely resemble human NIDDM. Genetic dissection with two kinds of F2 intercross progeny, from matings between the OLETF rat and non-diabetic control rats F344 or BN, allowed us to identify on Chromosome (Chr) 1 a major QTL associated with features of NIDDM that was common to both crosses. We also mapped two additional significant loci, on Chrs 7 and 14, in the (OLETF x F344)F2 cross alone, and designated these three loci as Diabetes mellitus, OLETF type Dmo 1, Dmo2 and Dmo3 respectively. With regard to suggestive QTLs, we found loci on Chrs 10, 11, and 16 that were common to both crosses, as well as loci on Chrs 5 and 12 in the (OLETF x F344)F2 cross and on Chrs 4 and 13 in the (OLETF x BN)F2 cross. Our results showed that NIDDM in the OLETF rat is polygenic and demonstrated that different genetic backgrounds could affect "fitness" for QTLs and produce different phenotypic effects from the same locus.

[A case of papillary fibroelastoma of the heart found 13 years after open mitral commissurotomy].

A patient, 68-year-old female, underwent open mitral commissurotomy (OMC) for rheumatic mitral stenosis in April, 1979. In July of 1992, during the postoperative follow-up echocardiogram showed 8 x 7 mm mitral subvalvular tumor mass and mitral restenosis. Then, mitral valve replacement was performed using prosthetic valve (Carbomedics #25 mm). The tumor was located on the top of postero-medial papillary muscle and its chordae, and histologically was diagnosed as benign papillary fibroelastoma. Based on the findings of periodic echocardiogram, the tumor doubling time was estimated to be 283 days. The postoperative course was uncomplicated and she was discharged on the 34th post operative day. Papillary fibroelastoma is a rare benign cardiac tumor. Pathological origin of this tumor is still unknown. However, Lambl's excrescence, long-term inflammatory stimulation and/or thrombi are supposed to be etiologic factors. This tumor is generally found incidentally at the time of autopsy or operation because the patients are often asymptomatic. However, because the tumor causes embolism at times, periodic examination by echocardiogram after operation preserving valves is recommended, and surgical resection is required when the diagnosis is established.

[A case of successfully operated hypertrophic obstructive cardiomyopathy in a child].

A case of surgical treatment of hypertrophic obstructive cardiomyopathy (HOCM) in a 5-year-old boy is reported. This is the youngest case in Japan. He had been diagnosed as HOCM and treated conservatively with beta-blocker and Ca-antagonist, but his complaints of dyspnea and chest oppression during efforts had increased. Cardiac catheterization showed a peak systolic pressure gradient of 110 mmHg between the left ventricle and systemic artery, and echocardiogram disclosed a systolic anterior motion (SAM) of the mitral valve and mitral regurgitation (MR). At operation transaortic septal myectomy and myotomy were performed according to preoperative findings by echocardiogram and MRI, and a marked reduction in pressure gradient and a decrease in SAM and MR were obtained. Thus, a satisfactory result can be obtained by myectomy and myotomy without mitral valve replacement in a pediatric patient with HOCM, based on the preoperative image analysis.

Determinants of myocardial oxygen consumption in fibrillating dog hearts. Comparison between normothermia and hypothermia.

The purpose of the present study was to elucidate the mechanism of the difference in myocardial oxygen consumption between heating and fibrillating states during normothermia and hypothermia. In five isolated cross-circulated dog hearts, we measured left ventricular pressure at several ventricular volumes and myocardial oxygen consumption at V0 and V100, at which peak isovolumic pressures were zero and approximately 100 mm Hg, respectively, in beating and fibrillating states during normothermia and hypothermia (29 degrees C). As a measure of the total mechanical energy at V100, we obtained pressure-volume area in the beating state and equivalent pressure-volume area for fibrillation. We calculated equivalent heart rate as an estimate of the contraction frequency of individual myocytes in a fibrillating ventricle from myocardial oxygen consumption at V0 in the beating and fibrillating states. During normothermia, myocardial oxygen consumption per minute at V0 and V100 and myocardial oxygen consumption for mechanical purposes at V100 (myocardial oxygen consumption at V100-myocardial oxygen consumption at V0) were significantly higher during fibrillation than in the beating state. Equivalent pressure-volume area during fibrillation and pressure-volume area in the beating state at V100 were comparable, whereas equivalent heart rate during fibrillation was significantly higher than heart rate in the beating state. During hypothermia, myocardial oxygen consumption was comparable between beating and fibrillating states at V0, although myocardial oxygen consumption at V100 was slightly lower during fibrillation than in the beating state. Myocardial oxygen consumption for mechanical purposes during fibrillation was half of that in the beating state. Equivalent pressure-volume area was significantly smaller than pressure-volume area, whereas equivalent heart rate and heart rate were comparable. We conclude that during normothermia, higher myocardial oxygen consumption during fibrillation than in the beating state at V0 and V100 is attributable to the higher contraction frequency. During hypothermia the comparable myocardial oxygen consumption values at V0 are attributable to the comparable contraction frequencies, whereas slightly lower myocardial oxygen consumption during fibrillation at V100 is ascribed to the lower total mechanical energy.

[A successful correction of common atrium with left atrial isomerism].

A successfully treated case of one-year-old girl of common atrium associated with bilateral superior vena cava and an interrupted inferior vena cava with hemiazygos continuation is reported. In addition to these anomalies of systemic venous connection, both atrial appendages show morphological characteristics of left type, and the final diagnosis of left atrial isomerism was made. In the corrective surgery, atrial septation was reconstructed using trimmed autogenous pericardial patch to allow the left superior vena cava to drain into the right side of the common atrium. The patient had been pointed out bradycardia (50-100/min) since her birth. Postoperative electrocardiogram also showed bradycardia (78-100/min). Especially in sleeping, her heart rate was 50-60/min, resulting in A-V junctional escaped rhythm and A-V dissociation. This fact suggests that her sinus node is absent or hypoplastic, one of the characteristics of left atrial isomerism. Since left atrial isomerism is reported to cause a decrease of atrial rhythm with aging, careful postoperative observation on specific arrhythmia is necessary.

New assist method for orthotopic heart transplantation in the dog.

We used cross circulation as an assist method for orthotopic heart transplantation in eight dogs. Seven of the eight dogs could be extubated on the first to third postoperative day and were able to eat and walk several hours after extubation. The survival times and the causes of death in these eight dogs were as follows: 436 days (alive), 5 days (sudden death), 17 days (rejection), 17 days (pyothorax), 3 days (accidental death), 59 days (rejection), 19 days (pneumothorax), and 188 days (alive). None of the dogs died of any problems related to the use of cross circulation. Hematologic and biochemical studies revealed that the CK, CK-MB, LDH, GOT, GPT, and lactate levels increased shortly after the operation and returned to the preoperative values within 14 days. The blood urea nitrogen, creatinine, amylase, hematocrit, and total protein levels remained almost unchanged. No free hemoglobin was detected. These findings indicated that no severe damage occurred to the recipient's organs. The key factors for performing successful cross circulation are as follows: (1) The body weight of the support dog should be at least three times that of the recipient. (2) The systolic pressure of the support dog and the pressure gradient between the support and the recipient should be kept above 150 mm Hg and 50 mm Hg, respectively. (3) The flow rate in the arterial tube should be maintained at between 30% and 50% of the basal cardiac output of the recipient. In conclusion, cross circulation is a suitable assist method for orthotopic heart transplantation in the dog.

[Cross circulation (CC) as an assist method for canine orthotopic cardiac transplantation--estimation of usefulness judging from intra- and postoperative hematological and biochemical examinations].

We employed cross circulation (CC) as an assist method for canine orthotopic cardiac transplantation in 8 dogs to obtain long term survivals. Hematological and biochemical examinations were performed intra- and postoperatively, to estimate usefulness of CC. The support dog weighted about 3 times as much as the recipient dog. Using different anesthesia, the systolic pressure of the support dog and the pressure gradient between the support and recipient were kept above 150 mmHg and 50 mmHg, respectively. Cyclosporin and prednisolone were administered for immunosuppressive therapy. All could be extubated on the 1st to 3rd postoperative day, and 6 survived more than 2 weeks. During the operation under CC, changes of LDH and lactic acid were much less, no free hemoglobin was detected, and total protein and hematocrit remained well compared with operation under heart-lung machines. CRK, CPKMB, GOT and GPT increased shortly after operation, but returned to the preoperative values within 2 weeks. In CC, owing to avoidance from mechanical destruction of red blood cells, coagulation disorders and hemodilution, hemolysis and hemorrhage could be decreased, and oncotic pressure remained well, and no severe damage occurred in the recipient organs. Thus, CC is useful as an assist method for canine orthotopic cardiac transplantation.

[A case of "chronic expanding hematoma", with an intrapulmonary fresh hemorrhage resected 25 years after thoracic injury].

A case of a 60-year-old man who had a chronic expanding hematoma with an intrapulmonary fresh hemorrhage in the right thorax is reported. He suffered blunt trauma of his right thorax in a traffic accident 25 years ago. He has no symptoms and signs after the accident. But he had hemoptysis twice and he was taken to our hospital. He underwent the resection of the chronic expanding hematoma and right lower lobe of the lung. The surgical specimen consisted of the hematoma measuring 17 x 13 x 6 cm. The chronic expanding hematoma contained aseptic brownish muddy substances. The patient recovered without complications and hemoptysis. Chronic expanding hematomas occur in many locations, often simulating neoplasms. Although the mechanism of chronic expanding hematoma is unclear, the self-perpetuating expanding nature of the lesion may appear to be due to the irritant effects of blood and its breakdown products, causing repeated exudation or bleeding from capillaries in the granulation tissue.

[A case of re-operation of pulmonary venous obstruction following surgical repair of total anomalous pulmonary venous return].

A case of six-month old female with pulmonary venous obstruction (PVO) following surgical repair of total anomalous pulmonary venous return (TAPVR) is reported. A digital subtraction angiography (DSA) performed at 9 days after birth showed a supracardiac type of TAPVR (Darling Ia) with pulmonary venous stenosis and pulmonary hypertension. On the following day she underwent a corrective operation through posterior approach. The anastomosis between common pulmonary venous chamber and left atrium had released the pulmonary venous stenosis. Nevertheless, the pulmonary hypertension progressed after the operation. Five months later a cardiac catheterization data showed PVO. Six months after the first operation she underwent a corrective re-operation for release from the PVO using EPTFE (expanded polytetrafluoroethylene) patch. Postoperatively the symptoms caused by PVO and pulmonary hypertension were markedly improved.

Successful correction of a Collett Edwards type II truncus arteriosus after bilateral pulmonary artery banding--a case report.

A case of a 4-year-old girl who underwent successful correction of a Collett Edwards type II truncus arteriosus following bilateral pulmonary artery banding is reported herein. The diagnosis of Collett Edwards type II truncus arteriosus was made when she was 5 months of age, after which the bilateral pulmonary artery banding was performed. This proved very effective and enabled elective radial surgery to be done. The radical operation was performed safely by a modified version of Rastelli's procedure. The patient had an uneventful recovery and is now well with no symptoms one year following her operation.