Aberrant activation of ALK kinase by a novel truncated form ALK protein in neuroblastoma.

Anaplastic lymphoma kinase (ALK) was originally identified from a rare subtype of non-Hodgkin's lymphomas carrying t(2;5)(p23;q35) translocation, where ALK was constitutively activated as a result of a fusion with nucleophosmin (NPM). Aberrant ALK fusion proteins were also generated in inflammatory fibrosarcoma and a subset of non-small-cell lung cancers, and these proteins are implicated in their pathogenesis. Recently, ALK has been demonstrated to be constitutively activated by gene mutations and/or amplifications in sporadic as well as familial cases of neuroblastoma. Here we describe another mechanism of aberrant ALK activation observed in a neuroblastoma-derived cell line (NB-1), in which a short-form ALK protein (ALK(del2-3)) having a truncated extracellular domain is overexpressed because of amplification of an abnormal ALK gene that lacks exons 2 and 3. ALK(del2-3) was autophosphorylated in NB-1 cells as well as in ALK(del2-3)-transduced cells and exhibited enhanced in vitro kinase activity compared with the wild-type kinase. ALK(del2-3)-transduced NIH3T3 cells exhibited increased colony-forming capacity in soft agar and tumorigenicity in nude mice. RNAi-mediated ALK knockdown resulted in the growth suppression of ALK(del2-3)-expressing cells, arguing for the oncogenic role of this mutant. Our findings provide a novel insight into the mechanism of deregulation of the ALK kinase and its roles in neuroblastoma pathogenesis.

Method to improve cosmetic outcome following craniotomy.

This technical note describes a simple method for reducing the dead space created by craniotome due to the loss of bone dust and improving the cosmetic outcome following a craniotomy. After drilling the burr holes for the craniotomy, the bone between the holes is drilled away in a standard fashion except that multiple regions of about 1 cm in length are left intact. These intact regions are broken using a periosteal elevator and fixed like a bridge when the bone is replaced. The resulting bone flap is readily returned to its original position without making the dead space created by regular craniotomy. The amount of the dead space caused by losing the bone dust is reduced and a good cosmetic recovery is obtained. This technique is useful for both craniotomy and facial bone surgery, which requires cosmetic results.

Changes in serum hypoxanthine levels after walk loads at mild to high intensity in healthy humans.

Effect of mild intensity exercise on the serum levels of hypoxanthine was studied. Eighteen healthy subjects performed 2 to 4 bouts of 5 minutes walk load at different intensities. At the beginning, thirteen of them walked at intensity more than 80% of the maximum. The serum levels of hypoxanthine increased to the levels of more than 6 times of resting values showing a peak at 10 to 20 minutes after the completion of the walk load. In 62 bouts of the walk load by 18 subjects, statistically significant relationship was demonstrated between intensity of the walk load and increase in serum concentration of hypoxanthine at 10 minutes after the completion of the walk load with correlation coefficient of 0.556. The serum hypoxanthine levels were significantly increased by the walk load even at mild intensity between 41 and 60%. Increment in the serum hypoxanthine concentration also showed positive and statistically significant correlation with physiological cost index. These results suggest that the serum levels of hypoxanthine increase following mild as well as moderate to submaximal intensity of exercise, and its increment may be used as an indicator of energy balance in the muscle during exercise at mild to high intensity.

Relation between changes in serum hypoxanthine levels by exercise and daily physical activity in the elderly.

Effect of exercise at mild intensity on the serum levels of hypoxanthine was studied in eleven healthy elderly subjects. They were divided into the active and sedentary groups according to their daily physical activity. They performed exercise testing to walk for 5 minutes keeping heart rate at approximately 70% of the maximum heart rate. Mean intensity of exercise estimated according to Karvonen's formula in the active or sedentary group was 41.8 +/- 9.6% or 34.1 +/- 6.1%, respectively. In the sedentary group, the serum hypoxanthine levels at 10 minutes after completion of walk load was significantly higher than that before exercise. Changes in the serum hypoxanthine levels in the active and sedentary groups were -0.97 +/- 1.36 and 0.80 +/- 0.57 micromol/liter, respectively (p < 0.05). This result suggests that mild intensity exercise increases the serum hypoxanthine concentration in the elderly leading inactive daily life, and physical activity suppresses an increase in the serum hypoxanthine levels by mild exercise.

Physical and psychological improvements after phase II cardiac rehabilitation in patients with myocardial infarction.

We have designed a new 4-week hospitalized phase II cardiac rehabilitation program. The purpose of the present study is to clarify whether the physical and psychological status of patients with myocardial infarction (MI) improves after participation in our program. Twenty-nine patients (27 males, two females) with acute MI who enrolled in the 4-week hospitalized phase II rehabilitation program were assessed. All patients enrolled in this study had received coronary interventions. The rehabilitation consisted of exercise training, education and counseling. We evaluated the physical and psychological status of the patients before and just after the program, and at a 6-month follow up. The physical status was assessed by exercise tolerance measured by the peak oxygen consumption and anaerobic threshold, frequency of exercise, and serum concentrations of triglyceride, total cholesterol, high-density lipoprotein-cholesterol, and low-density lipoprotein-cholesterol. The psychological status was assessed by the Spielberger state-trait anxiety inventory questionnaire (STAI) and the self-rating questionnaire for depression (SRQ-D). Thirty-four patients (27 men, seven women) with MI who did not participate in our rehabilitation program served as a control group. After participation in our rehabilitation program, exercise tolerance and the serum lipid profiles of the patients were improved compared with those before rehabilitation. These parameters had improved significantly 6 months after rehabilitation. The STAI anxiety score was improved significantly and the SRQ-D depression score tended to be improved just after the rehabilitation program. Regular physical activity was continued even 6 months after the completion of the program. Our hospitalized phase II cardiac rehabilitation program improved the management of cardiac risk factors and the psychological status in patients with MI. This comprehensive program may contribute to the secondary prevention of MI as well as the recovery of physical and psychological activities.

A clinical variance in familial amyotrophic lateral sclerosis with a point mutation in Cu/Zn superoxide dismutase gene.

We report here a novel point mutation in exon 5 of the Cu/Zn superoxide dismutase (SOD) gene resulting in an amino acid substitution of valine(148) by isoleucine (V148I) in a Japanese family with amyotrophic lateral sclerosis (FALS). In this family, the age at onset was young (28.0 ± 3.8 years old, mean ± SD, n = 4) and the disease progression was rapid (22.0 ± 5.9 months, n = 3) with low Cu/Zn SOD activity (56.3 and 59.0% of the controls, n = 2). It is interesting that the clinical features of ALS varied very much among the affected members. One case had weakness of the lower extremities at first, and died without bulbar paresis. The second case first noticed wasting of the upper limbs with bulbar symptoms, but the third had weakness of upper extremities without developing dysarthria nor dysphagia until death. The living remainder first developed fasciculation of the tongue without weakness of extremities. The valine(148) is conserved among different species, and V148I mutation might destabilize dimer formation with another SOD subunit, leading to decrease enzymatic activity. These results suggested that there could be considerable clinical variance among the patients of FALS within one family, carrying the same Cu/Zn SOD mutation such as V148I.

Control of diabetes during rehabilitation for diabetic stroke.

Changes of diabetic control during rehabilitation therapy was studied retrospectively in 33 cases of diabetic stroke. Diabetic control was not deteriorated in any cases during rehabilitation therapy. In 21 cases, treatment of diabetes was not changed during rehabilitation therapy and the levels of fasting plasma glucose were decreased in all of them. Doses of hypoglycemic agents was reduced without significant deterioration of diabetic control in 9 cases whose diabetic control was excellent, and in 2 cases who experienced hypoglycemic attack during rehabilitation therapy. In these cases, oral hypoglycemic agent was reduced from 2 to 1 tablet and insulin from 32 to 21 units per day on average. These results indicate that training in the stroke rehabilitation moderately improves diabetic control.

Changes of plasma lipid and lipoproteins by rehabilitation therapy for stroke.

Changes of the plasma levels of lipid by rehabilitation therapy for approximately 10 weeks were studied in 14 cases of stroke. Hypercholesterolemia or hypertriglyceridemia was present in a half cases of stroke, and HDL cholesterol was lower than the normal level in most cases before starting rehabilitation therapy. After rehabilitation therapy, hypertriglyceridemia was normalized in most cases. On the other hand, the plasma levels of cholesterol and LDL cholesterol were decreased slightly. HDL cholesterol was increased and atherogenic index was improved by rehabilitation therapy.

Urinary active kallikrein excretion and diabetic renal impairments in streptozotocin-treated rats.

To assess possible roles of the renal kallikrein-kinin system in the development of renal impairments in diabetes mellitus, we determined daily excretion of urinary total and active kallikrein in uninephrectomized Wistar-Kyoto rats made diabetic by streptozotocin (45 mg/kg) as a bolus injection. We also evaluated the effect of captopril (50 mg/kg/day) administered orally on the development of diabetic renal impairments in the streptozotocin-treated rats. Active kallikrein was determined by its kininogenase activity, and generated kinins were radioimmunologically measured. Total kallikrein was also determined by measuring kininogenase activity after inactive kallikrein had been activated with trypsin (200 micrograms/ml). Urinary active kallikrein excretion was significantly reduced in streptozotocin-treated rats whereas urinary total kallikrein excretion was unchanged, resulting in the decreased ratio of active to total kallikrein compared to that in the controls. These reductions were preceded by the increased excretion of urine protein measured as an index of renal impairments. The administration of captopril for 12 weeks attenuated the development of diabetic renal impairments evaluated by urine protein excretion in streptozotocin-treated rats, although it did not induce significant changes in urinary total and active kallikrein excretion, and the ratio of active to total kallikrein. Thus the results of this study indicate that the renal kallikrein-kinin system might not play major roles in the development of diabetic renal impairments in the rat, although the pathophysiological relevance of impaired activation of renal kallikrein system to the development of diabetic renal impairments remains to be determined. In addition, they suggest that the renoprotective effects of captopril may be independent of the activation of renal kallikrein system in streptozotocin-treated rats.

The augmented blood pressure response to chronic angiotensin II infusion in partially nephrectomized rats.

To assess potential roles of circulating levels of angiotensin II (Ang II) in the regulation of blood pressure in rats with reduced functional renal mass, we studied the blood pressure response to chronic infusion of Ang II at a subpressor dose in partially nephrectomized rats. Male Sprague-Dawley rats were subjected to 1/6, 2/6, 3/6, 4/6 and 5/6 nephrectomy, and sham operation a week before the infusion of Ang II at a rate of 400 micrograms/kg/day delivered intraperitoneally by osmotic minipumps for 14 days, or vehicle alone served as controls. Infusion of Ang II was associated with a slight but significant increase in systolic blood pressure in 1/6 nephrectomized rats (p < 0.05), whereas it did not induce a significant change in systolic blood pressure in sham-operated rats. Hypertension during Ang II infusion was more prominent in 2/6 nephrectomized rats than that in 1/6 nephrectomized rats (p < 0.05). However, there was no significant difference in hypertension during Ang II infusion among groups in 2/6, 3/6, 4/6 and 5/6 nephrectomized rats. Blood urea nitrogen and serum creatinine levels, endogenous creatinine clearance, urinary excretion of protein as indices of renal function, and urinary sodium excretion were not affected by chronic infusion of Ang II in 1/6, 2/6, 3/6, 4/6 and 5/6 nephrectomized, and sham-operated rats. There was also no significant difference in circulating levels of Ang II among Ang II infused-groups in 1/6 and 2/6 nephrectomized rats. These results indicate that elevated levels of circulating Ang II are capable of causing a sustained hypertension in rats with reduced functional renal mass.(ABSTRACT TRUNCATED AT 250 WORDS)

Chronic treatment with anti-endothelin antibodies fails to modify the development of hypertension in stroke-prone spontaneously hypertensive rats and DOCA-salt hypertensive rats.

This study was designed to assess whether blocking endogenous endothelin with anti-endothelin antibodies could alter the development of hypertension in stroke-prone spontaneously hypertensive rats (SHR) and DOCA-salt treated rats. Specific anti-endothelin antibodies were produced in rabbits by standard methods. The amount of anti-endothelin antibodies employed in this study blocked the hypertensive effect of endothelin-1, 750 ng/kg, by 55% in conscious rats. Intravenous injection of anti-endothelin antibodies as a bolus twice a week for 3 weeks did not affect the rise in blood pressure of stroke-prone SHR (268 +/- 8 mmHg, n = 8) compared to control stroke-prone SHR (256 +/- 7 mmHg, n = 8) treated with normal rabbit serum. Intravenous administration of anti-endothelin antibodies in a same manner also failed to alter the development of hypertension in DOCA-salt treated rats (160 +/- 6 mmHg in anti-endothelin antibodies-treated group, n = 7 compared to 164 +/- 5 mmHg in normal rabbit serum-treated group, n = 7). The administration of anti-endothelin antibodies did not induce any significant changes in body weight, urine volume and urinary sodium excretion in stroke-prone SHR and DOCA-salt treated rats compared to those treated with normal rabbit serum. These findings suggest that circulating endothelin might not play a major role in the regulation of blood pressure in stroke-prone SHR and DOCA-salt treated rats.

Antihypertensive effect of cilazapril in essential hypertensive patients. Clinical study with 24 h ambulatory blood pressure monitoring.

In the present study, the efficacy of cilazapril (Ro 31-2848/006, CAS 88768-40-5), a new angiotensin converting enzyme (ACE) inhibitor, was examined in 7 essential hypertensive patients by monitoring ambulatory blood pressure for 24 h. Oral cilazapril, given once a day in the morning, lowered systolic and diastolic blood pressure without affecting heart rate. Daily profiles of blood pressure and heart rate, however, were not different from those prior to cilazapril in these patients. Percentages of high blood pressure values (more than 160/95 mmHg), both systolic and diastolic decreased, following cilazapril administration. Ambulatory blood pressure monitoring revealed that cilazapril did not cause a dangerous fall in blood pressure even in the night time. From these results, cilazapril could be an effective and useful antihypertensive drug for the treatment of essential hypertensive patients.

Antihypertensive effect of captopril and enalapril in endothelin-infused rats.

Comparative effects of angiotensin converting enzyme inhibitors and calcium channel blockers were assessed in rats infused chronically with synthetic endothelin. When 50 mg/kg/day of captopril orally or 6 mg/kg/day of enalapril intraperitoneally was administered simultaneously with 60 micrograms/kg/day of endothelin, the systolic blood pressure was on Day 1 142.7 +/- 5.9 mmHg (p less than 0.05) or 128.7 +/- 6.7 mmHg (p less than 0.05), respectively, compared to the rise to 163.8 +/- 4.7 mmHg when endothelin alone was infused. The antihypertensive effect of captopril or enalapril was sustained for the entire experimental period and was not associated with a significant change in urinary sodium excretion, whereas both drugs induced a significant increase in urine volume. Chronic infusion of angiotensin II intraperitoneally at a subpressor dose (400 micrograms/kg/day) reversed the antihypertensive effect of captopril in endothelin-infused rats. When 6 mg/kg/day of benidipine or 10 mg/kg/day of nilvadipine orally was administered simultaneously with 60 micrograms/kg/day of endothelin, the systolic blood pressure was on Day 1 137.0 +/- 2.4 mmHg (p less than 0.05) or 119.7 +/- 5.9 mmHg (p less than 0.05), respectively, compared to the rise when endothelin alone was infused. The antihypertensive effect of benidipine or nilvadipine was sustained for the entire experimental period and was not associated with any significant changes in urine volume and urinary sodium excretion. These results indicate that the reduced sensitivity of the peripheral arteries to endothelin may be involved in the mechanism of the hypotensive action of angiotensin converting enzyme inhibitors, dependent on the suppressed angiotensin II formation.