Cytochemical analysis of storage materials in cultured skin fibroblasts from patients with I-cell disease

BACKGROUND: In cultured fibroblasts from I-cell disease patients the transport of many lysosomal enzymes is defective, and affected cells contain inclusion bodies filled with undegraded substrates. However, the contents of these inclusion bodies have not been well characterized yet. We attempted to identify accumulated substances in cultured I-cell disease fibroblasts cytochemically. METHODS: Cultured fibroblasts from I-cell disease patients were double-stained with a monoclonal antibody to lysosome-associated membrane protein-1 (LAMP-1) and that to GM2 ganglioside, or a series of lectins that specifically bind to sugar moieties. RESULTS: The patients' cells were granularly stained with the antibody to GM2 ganglioside and the lectins including Maakia amurensis, Datura stramonium, and concanavalin A. Their localization was coincident with that of LAMP-1. CONCLUSIONS: GM2 ganglioside and various kinds of glycoconjugates having sialic acidalpha2-3galactose, galactosebeta1-4N-acetylglucosamine and mannose residues accumulate in enlarged lysosomes in I-cell disease fibroblasts.     

Detection of thrombosis in a magnetically levitated blood pump by vibrational excitation of the impeller

The use of contactless support technology for the impeller has led to an increase in the durability of ventricular assist devices (VADs), and these have been in clinical use worldwide. However, pump thrombosis and stroke are still issues to be solved. We have developed a method for detecting the thrombosis in a magnetically levitated blood pump without the need for additional sensors or other equipment. In the proposed method, a sinusoidal current is applied to the electromagnets used for the magnetic bearing, resulting in vibration of the impeller. The phase difference between the current and displacement of the impeller increases with pump thrombosis. First, we describe the principle by which the pump thrombosis is detected. Pump thrombosis reduces the narrowest fluid gap in the pump and this gives rise to a change in the phase difference. Second, we report on experiments in which we changed the narrowest fluid gap using oriented polypropylene tape and showed that decreasing the narrowest fluid gap resulted in an increase in phase difference. For these experiments, the measurements were repeated three times for each condition. Third, we examine the relationship between the pump thrombosis and the phase difference evaluated by observations of the underside of the impeller when operating the pump with porcine blood. Since light was unable to penetrate the blood layer, the erythrocytes were removed for this observation. Only one observation was made. The results showed the phase difference rapidly increased at the same moment when the pump thrombosis was observed. This implies the proposed method has the potential to detect the early stages of pump thrombosis. Finally, in vitro experiments to detect thrombosis when using whole porcine blood in the pump were conducted. The experiment was carried out five times. To intentionally form a thrombus inside the pump, the activated clotting time was controlled to be less than 200 s. In every case, the phase difference increased by more than one degree after tens of minutes. Then, the pump was disassembled and a small amount of pump thrombosis was observed. We conclude that real-time diagnosis of pump thrombosis may be realized by measuring the phase difference without the need for additional sensors.     

Species identification of Asini Corii Collas (donkey glue) by PCR amplification of cytochrome b gene

Asini Corii Collas (ACC; donkey glue) is a crude drug used to promote hematopoiesis and arrest bleeding. Because adulteration of the drug with substances from other animals such as horses, cattle, and pigs has been found, we examined PCR methods based on the sequence of the cytochrome b gene for source species identification. Two strategies for extracting DNA from ACC were compared, and the ion-exchange resin procedure was revealed to be more suitable than the silica-based one. Using DNA extracted from ACC by the ion-exchange resin procedure, PCR methods for species-specific detection of donkey, horse, cattle, and pig substances were established. When these species-specific PCR methods were applied to ACC, amplicons were obtained only by the donkey-specific PCR. Cattle-specific PCR detected as little as 0.1 % admixture of cattle glue in the ACC. These results suggest that the species-specific PCR methods established in this study would be useful for simple and easy detection of adulteration of ACC.     

Range of motion of thoracic spine in sagittal plane

Study design

Imaging study of thoracic spine.

Objective

The purpose of this study was to investigate dynamic alignment and range of motion (ROM) at all segmental levels of thoracic spine.

Summary of background data

Thoracic spine is considered to have restricted ROM because of restriction by the rib cage. However, angular movements of thoracic spine can induce thoracic compressive myelopathy in some patients. Although few previous studies have reported segmental ROM with regard to sagittal plane, these were based on cadaver specimens. No study has reported normal functional ROM of thoracic spine.

Methods

Fifty patients with cervical or lumbar spinal disease but neither thoracic spinal disease nor compression fracture were enrolled prospectively in this study (34 males, 16 females; mean age 55.4 ± 14.7 years; range 27–81 years). After preoperative myelography, multidetector-row computed tomography scanning was performed at passive maximum flexion and extension position. Total and segmental thoracic kyphotic angles were measured and ROM calculated.

Results

Total kyphotic angle (T1/L1) was 40.2° ± 11.4° and 8.5° ± 12.8° in flexion and extension, respectively (P < 0.0001). The apex of the kyphotic angle was at T6/7 in flexion. Total ROM (T1/L1) was 31.7° ± 11.3°. Segmental ROM decreased from T1/2 to T4/5 but increased gradually from T4/5 to T12/L1. Maximum ROM was at T12/L1 (4.2° ± 2.1°) and minimum at T4/5 (0.9° ± 3.0°).

Conclusions

Thoracic spine showed ROM in sagittal plane, despite being considered a stable region. These findings offer useful information in the diagnosis and selection of surgical intervention in thoracic spinal disease.     

The mutations of the EGFR and K-ras genes in resected stage I lung adenocarcinoma and their clinical significance

Purpose

This study retrospectively assessed the mutations of the epidermal growth factor receptor (EGFR) and K-ras genes and their clinical significance in patients with resected stage I adenocarcinomas.

Methods

A total of 354 patients with resected lung adenocarcinomas were included, and 256 patients with stage I disease were analyzed for the prognostic and predictive value of these mutations.

Results

Mutations of EGFR and K-ras genes were detected in 149 (41.1 %) and 23 (6.4 %) of all tumors, and in 122 (47.6 %) and 14 (5.5 %) of stage I tumors, respectively. There were no significant differences in the disease-free survival (DFS) and overall survival (OS) between the EGFR-mutant and wild-type groups. However, the DFS and OS were significantly shorter in patients with K-ras mutations than in those without (5-year DFS: 50.8 vs. 76.9 %, 5-year OS: 70.0 vs. 86.6 %, p < 0.01). A multivariate analysis showed that K-ras mutations were an independent poor prognostic factor. Twenty-four of the 41 patients with recurrent disease after surgery were treated with an EGFR–TKI. Fifteen EGFR-mutant patients treated with an EGFR–TKI had a better prognosis than did the nine EGFR-wild-type patients.

Conclusion

The presence of an EGFR gene mutation was a predictive factor for the response to EGFR–TKI treatment in patients with resected stage I adenocarcinoma, but was not a prognostic factor. The presence of a K-ras gene mutation was a poor prognostic factor.