Novel TENM3-ALK fusion is an alternate mechanism for ALK activation in neuroblastoma.

The identification of molecular events underlying the pathogenesis of neuroblastoma can likely result in improved clinical outcomes for this disease. In this study, a translocation within chromosome 2p and 4q was found to bring about the formation of an in-frame fusion gene that was composed of portions of the teneurin transmembrane protein 3 (TENM3, also known as ODZ3) gene and the anaplastic lymphoma kinase (ALK) gene in tumor cells from patients with neuroblastoma. Expression of the full length TENM3-ALK cDNA in NIH-3T3 cells led to the formation of a fusion protein that: (1) possesses constitutive tyrosine kinase activity, (2) induces strong activation of the downstream targets of extracellular signal-regulated kinase (ERK), protein kinase B (a.k.a. AKT), and signal transducer and activator of transcription 3 (STAT3), (3) provokes oncogenic transformation in NOD.Cg-PrkdcscidIl2rgtm1Sug/ShiJic mice, and (4) possesses sensitivity to ALK inhibitors in vitro and in vivo. Our findings demonstrated that patients with neuroblastoma may express a transforming fusion kinase, which is a promising candidate for a therapeutic target and a diagnostic molecular marker for neuroblastoma. The in-frame 5' partner gene that fuses with ALK has not been reported previously in neuroblastoma. Our data provide novel biological insights into the mechanism of ALK activation due to translocation, with implications for neuroblastoma tumorigenesis, and could be useful as a vital marker for the accurate diagnosis of this type of neuroblastoma.

Rapid progression to end-stage renal disease in a child with IgA-dominant infection-related glomerulonephritis associated with parvovirus B19.

Parvovirus B19 (PVB19) has been known to cause acute glomerulonephritis and nephrotic syndrome with various renal histologic patterns, such as endocapillary glomerulonephritis and collapsing glomerulopathy. Remission is achieved spontaneously or by treatment with steroid and/or immunosuppressants in most patients, except those with sickle cell anemia or two APOL1 risk alleles. In this study, we report the case of a previously healthy 5-year-old boy with infection-related glomerulonephritis (IRGN) associated with PVB19 that progressed to end-stage renal disease (ESRD). He presented with macrohematuria, nephrotic-range proteinuria, and progressive renal dysfunction despite treatment with methylprednisolone pulse therapy, plasmapheresis, and intravenous immunoglobulin. The kidney biopsy specimens exhibited endocapillary infiltration and mesangiolysis with cellular crescent formation. Immunofluorescence analysis revealed that IgA was dominantly positive in the glomeruli, with some co-localized with KM55, which is a specific monoclonal antibody for galactose-deficient IgA1 (Gd-IgA1). The intensity of the KM55 signal in the present patient was weaker than that in patients with IgA nephropathy. To our knowledge, this is the first report of IRGN associated with PVB19 that progressed to ESRD without any underlying diseases. Further investigations are needed to determine the significance of IgA and Gd-IgA1 deposition in IRGN associated with PVB19.

Long-term outcome of renal transplantation in childhood-onset anti-neutrophil cytoplasmic antibody-associated vasculitis.

BACKGROUND: There have been a few reports of RTx for AAV in children; however, post-transplant recurrence rate and long-term prognosis remain unclear. Here, we describe the long-term outcomes of RTx in childhood-onset AAV. METHODS: We conducted a retrospective study of children who underwent RTx for AAV between 1999 and 2017 and had a follow-up period of >2 years. RESULTS: Seven patients consisting of three children with MPA and four with RLV were analyzed. Age at Dx was 5.9 (median; range, 4.1-14.5) years. PD was instituted in all patients, and median time on dialysis was 26 (range, 14-63) months. Age at RTx was 12.8 (median; range, 8.7-16.3) years. There were no recurrences of AAV noted during the median follow-up period of 7.0 (range, 2.7-18.8) years after RTx. Graft loss occurred in one patient due to non-adherence. Estimated glomerular filtration rate of the remaining patients at the last follow-up was 73.0 (median; range, 50.7-93.9) mL/min/1.73 m2 . No malignancies and deaths occurred during the observational period. CONCLUSIONS: Our study suggests that RTx for AAV with ESRD is a potentially safe and effective treatment choice for children with AAV.

Clinical characteristics of Campylobacter enteritis after pediatric renal transplantation: A retrospective analysis from single center.

BACKGROUND: There are few reports of patients with Campylobacter enteritis after renal transplantation, and only a few case reports of bacteremia have been published. Although antibiotic therapy for 3-5 days has been recommended for immunocompromised patients, the optimal treatment for Campylobacter enteritis after renal transplantation has not been established. This study aimed to clarify the clinical characteristics and treatment outcomes of Campylobacter enteritis after pediatric renal transplantation. METHODS: This retrospective study included patients who underwent pediatric renal transplantation and were found to have Campylobacter species in stool cultures between January 2014 and May 2017. RESULTS: This study included eight patients who underwent pediatric renal transplantation. The median age at the time of renal transplantation was 14 years, and the median period between transplantation and disease occurrence was 4.6 years. Clinical symptoms were abdominal pain for eight patients, diarrhea for eight patients, fever for seven patients, vomiting for three patients, and headache for three patients. Campylobacter jejuni was isolated from the stool cultures of all patients. The median administration period of antibiotics as initial therapy was 7 days (range, 4-11 days). However, clinical relapse was observed in four patients after completing antibiotic therapy. Patients who experienced clinical relapse required a second course of antibiotic therapy for a median duration of 7 days (range, 5-10 days). CONCLUSIONS: Patients with Campylobacter enteritis after pediatric renal transplantation are at high risk for clinical relapse and may require a longer duration of antibiotic therapy than that generally described.

Slower recovery rate of muscle oxygenation after sprint exercise in long-distance runners compared with that in sprinters and healthy controls.

The purpose of this study was to examine whether differences in aerobic capacity and training status influence muscle reoxygenation after sprint exercise. We hypothesized that the muscle reoxygenation rate after sprint exercise is slower in long-distance runners with a high aerobic capacity. Five male long-distance runners, 5 male sprinters, and 6 healthy male controls performed a 30-second sprint exercise on a cycle ergometer. Oxygen saturation in muscle tissue (StO2) in the vastus lateralis muscles was measured by near-infrared spectroscopy. The muscle reoxygenation rate after the exercise was evaluated at half the time required for StO2 recovery (T1/2 StO2). Aerobic capacity was evaluated by measuring maximal oxygen consumption (V[Combining Dot Above]O2max). The T1/2 StO2 in the long-distance runners (25.0 ± 4.5 seconds) was significantly longer than that in the controls (15.9 ± 1.6 seconds; p < 0.01) and in the sprinters (18.0 ± 4.6 seconds; p < 0.05). In all the subjects (long-distance runners, sprinters, and controls), the T1/2 StO2 had a significant positive correlation with the V[Combining Dot Above]O2max (r = 0.75; p < 0.01) and was longer in subjects with a higher V[Combining Dot Above]O2max. These results suggest that reoxygenation after sprint exercise is influenced by aerobic capacity and training status, and that the subjects with a higher aerobic capacity have delayed muscle reoxygenation after sprint exercise.

Effect of exercise intensity on oxygen consumption kinetics in non-exercising muscle during exercise.

This study examined the effect of exercise intensity on the kinetics of muscle oxygen consumption in non-exercising forearm flexor muscles (VO(2mf)) during exercise. Seven healthy male subjects performed cycling exercise for 60 min at 30% of maximal oxygen consumption (%VO(2max)) and 30 min at 50% VO(2max) on separate days. The VO(2mf) values at rest and during exercise were measured by near-infrared spectroscopy. The VO(2mf) at 30% VO(2max) significantly increased to 1·2 ± 0·1-fold over resting value at 20 min after the beginning of exercise (P<0·05) and remained constant within 1·2- to 1·3-fold over resting value until 60 min during exercise. The VO(2mf) at 50% VO(2max) significantly increased to 1·2 ± 0·1-fold over resting value at 15 min after the beginning of exercise (P<0·05). Subsequently, the VO(2mf) at 50% VO(2max) increased with time to 1·3 ± 0·1-fold over resting value at 20 min after the beginning of exercise and to 1·5 ± 0·2-fold over resting value at 30 min. The VO(2mf) 15-30 min of exercise at 50% VO(2max) was significantly higher than that at 30% VO(2max) (P<0·05). These data suggest that the increase in VO(2mf) has a time lag from the beginning of exercise, and the kinetics of VO(2mf) during exercise differs with exercise intensity. Therefore, we conclude that the kinetics of VO(2mf) during exercise is dependent on exercise intensity.

Resistance exercise increases postexercise oxygen consumption in nonexercising muscle.

This study examined the effect of knee extension resistance exercise on muscle oxygen consumption in nonexercising forearm flexor muscles (nonexV(O)(2mus)) after exercise. Seven healthy male subjects were performed six sets of unilateral knee extension exercise until exhaustion at 40, 60, and 80% of 1 repetition maximum (RM) on separate days. The nonexV(O)(2mus) values at rest, at the end of exercise, and during recovery after exercise were measured by near-infrared spectroscopy. The nonexV(O)(2mus) at the end of exercise was significantly (P < 0.05) increased by 1.8 +/- 0.2, 1.7 +/- 0.2, and 1.4 +/- 0.3 fold over resting value at 40, 60 and 80% 1RM, respectively. NonexV(O)(2mus) returned to the resting values after 1-5 min of recovery and then showed no further significant change for all exercise intensities. This study suggests that knee extension resistance exercise at 40, 60 and 80% 1RM induced an increase in nonexV(O)(2mus) and that the increase of nonexV(O)(2mus) after exercise returned to resting value in several minutes.

Role of disulphide bonds in a thermophilic serine protease aqualysin I from Thermus aquaticus YT-1.

A thermophilic serine protease, Aqualysin I, from Thermus aquaticus YT-1 has two disulphide bonds, which are also found in a psychrophilic serine protease from Vibrio sp. PA-44 and a proteinase K-like enzyme from Serratia sp. at corresponding positions. To understand the significance of these disulphide bonds in aqualysin I, we prepared mutants C99S, C194S and C99S/C194S (WSS), in which Cys69-Cys99, Cys163-Cys194 and both of these disulphide bonds, respectively, were disrupted by replacing Cys residues with Ser residues. All mutants were expressed stably in Escherichia coli. The C99S mutant was 68% as active as the wild-type enzyme at 40 degrees C in terms of k(cat) value, while C194S and WSS were only 6 and 3%, respectively, as active, indicating that disulphide bond Cys163-Cys194 is critically important for maintaining proper catalytic site conformation. Mutants C194S and WSS were less thermostable than wild-type enzyme, with a half-life at 90 degrees C of 10 min as compared to 45 min of the latter and with transition temperatures on differential scanning calorimetry of 86.7 degrees C and 86.9 degrees C, respectively. Mutant C99S was almost as stable as the wild-type aqualysin I. These results indicate that the disulphide bond Cys163-Cys194 is more important for catalytic activity and conformational stability of aqualysin I than Cys67-Cys99.

Age and activity status affect muscle reoxygenation time after maximal cycling exercise.

PURPOSE: The purpose of this study was to determine the interaction of age and habitual physical activity on recovery time of muscle oxygenation following maximal cycling exercise (CycEXmax). METHODS: Twelve sedentary middle-aged (50+/-6), 13 sedentary elderly (66+/-3), 13 active middle-aged (53+/-5), and 20 active elderly (67+/-5) women participated in this study. We evaluated the peak pulmonary oxygen uptake (VO2peak) during CycEXmax and the half-recovery time of muscle oxygenation (T1/2reoxy time) using near-infrared spectroscopy at the vastus lateralis (VL) during the recovery phase after CycEXmax. RESULTS: T1/2reoxy time was significantly greater in the elderly subjects than in the middle-aged subjects in both sedentary (P<0.05) and active groups (P<0.01). T1/2reoxy time of the active group was lower (P<0.01) than that of the sedentary group regardless of age. Age was significantly correlated to T1/2reoxy time in both sedentary and active groups (in both sedentary and active groups: P<0.01). The slope of T1/2reoxy time against age in the sedentary group was significantly greater (VL: P<0.05) than that of the active group. VO2peak showed significant inverse correlation with T1/2reoxy time at the VL in both sedentary and active groups. The slope of VO2peak against T1/2reoxy time showed no significant differences between middle-aged and elderly subjects. CONCLUSION: The results of this study suggest that T1/2reoxy time was prolonged with aging, regardless of habitual physical activity levels. However, habitual physical activity may prevent the age-related prolongation in T1/2reoxy time after CycEXmax. VO2peak appears to be one of the major factors determining T1/2reoxy time, not age.

Significance of post-exercise increment of urinary bicarbonate and pH in subjects loaded with submaximal cycling exercise.

We studied the changes in urinary bicarbonate, urinary pH and some physical parameters such as minute ventilation (VE), oxygen consumption (VO2), respiratory carbon dioxide (VCO2), heart rate, blood pressure, and blood lactate, before and after the submaximal exercise. Six male subjects aged 28-33 years were involved in the study. They performed the incremental exercise test using a bicycle ergometer until exhaustion. Levels of VE, VO2, VCO2, heart rate, and blood pressure increased continuously with an increase in cycling intensity. These parameters markedly decreased and reached the baseline levels within 5-10 minutes after the termination of exercise. According to an increase in cycling intensity, blood lactate increased continuously during exercise, but after termination of exercise the return of lactate to the baseline level was markedly retarded. Urinary bicarbonate and pH were within the range of those at 0 time (baseline levels) from the beginning until 30 minutes after the exercise. However, they began to increase abruptly about 30 minutes after the exercise, and continued to increase extensively for 2 hours thereafter. Such marked increase in urinary bicarbonate and pH seemed to be correlated with the aerobic metabolism of lactate in the muscles, liver, and kidney, finally producing CO2. It was also suggested that the measurement of urinary bicarbonate and pH may be useful for the estimation of physiological changes in the body after submaximal incremental cycling exercise loading.

Noninvasive monitoring of deterioration in skeletal muscle function with forearm cast immobilization and the prevention of deterioration.

BACKGROUND: In this research inactivity was simulated by immobilizing the forearm region in a plaster cast. Changes in skeletal muscle oxidative function were measured using near-infrared spectroscopy (NIRS), and the preventative effect of the training protocol on deterioration of skeletal muscle and the clinical utility of NIRS were examined. METHODS: Fourteen healthy adult men underwent immobilization of the forearm of the non-dominant arm by plaster cast for 21 days. Eight healthy adult subjects were designated as the immobilization group (IMM) and six were designated as the immobilization + training group (IMM+TRN). Grip strength, forearm circumference and dynamic handgrip exercise endurance were measured before and after the 21-day immobilization period. Using NIRS, changes in oxidative function of skeletal muscles were also evaluated. Muscle oxygen consumption recovery was recorded after the completion of 60 seconds of 40% maximum voluntary contraction (MVC) dynamic handgrip exercise 1 repetition per 4 seconds and the recovery time constant (TcVO2mus) was calculated. RESULTS: TcVO2mus for the IMM was 59.7 +/- 5.5 seconds (average +/- standard error) before immobilization and lengthened significantly to 70.4 +/- 5.4 seconds after immobilization (p < 0.05). For the IMM+TRN, TcVO2mus was 78.3 +/- 6.2 seconds before immobilization and training and shortened significantly to 63.1 +/- 5.6 seconds after immobilization and training (p < 0.05). CONCLUSIONS: The training program used in this experiment was effective in preventing declines in muscle oxidative function and endurance due to immobilization. The experimental results suggest that non-invasive monitoring of skeletal muscle function by NIRS would be possible in a clinical setting.

Deterioration of muscle function after 21-day forearm immobilization.

PURPOSE: Although it is well known that immobilization causes muscle atrophy, most immobilization models have examined lower limbs, and little is known about the forearm. The purpose of this study was to determine whether forearm immobilization produces changes in muscle morphology and function. METHODS: Six healthy males (age: 21.5 +/- 1.4, mean +/- SD) participated in this study. The nondominant arm was immobilized with a cast (CAST) for 21 d, and the dominant arm was measured as the control (CONT). The forearm cross-sectional area (CSA) and circumference were measured as muscle morphology. Maximum grip strength, forearm muscle oxidative capacity, and dynamic grip endurance were measured as muscle function. Magnetic resonance (MR) imaging was used to measure CSA, and 31phosphorus MR spectroscopy was used to measure time constant (Tc) for phosphocreatine (PCr) recovery after submaximal exercise (PCr-Tc). Grip endurance was expressed by the number of handgrip contractions at 30% maximum grip strength load. All measurements were taken before and after the immobilization. RESULTS: After the 21-d forearm immobilization, no changes were seen for each measurement in CONT. CSA and the circumference showed no significant changes in CAST. However, maximum grip strength decreased by 18% (P < 0.05), PCr-Tc was prolonged by 45% (P < 0.05), and the grip endurance at the absolute load was reduced by 19% (P < 0.05) for CAST. CONCLUSION: In this model, 21-d forearm immobilization caused no significant changes in forearm muscle morphology, but the muscle function showed remarkable deterioration ranging from 18 to 45%.