Skeletal muscle expression of clathrin and mannose 6-phosphate receptor in experimental chloroquine-induced myopathy

Previous studies suggest that the muscle fiber lysosome system plays a central role in the increased formation of autophagosomes and autolysosomes that occurs in the context of chloroquine-induced myopathy. The goal of this study was to characterize the contribution of receptor-mediated intracellular transport, particularly the endosomal pathway, to the abnormal accumulation of vacuoles in experimental chloroquine myopathy. Expression of the mannose 6-phosphate receptor (M6PR) and clathrin were analyzed in innervated and denervated rat soleus muscles after treatment with either saline or chloroquine. Accumulation of vacuoles was observed only in chloroquine-treated denervated muscles. Further, clathrin immunostaining and M6PR messenger ribonucleic acid (mRNA) were significantly increased in denervated soleus muscle from saline- and chloroquine-treated rats compared to contralateral, innervated muscles. However, there was no difference in clathrin levels when comparing saline- and chloroquine-treated denervated muscles. These data suggest that chloroquine activates the transport of newly synthesized lysosomal enzymes from the secretory pathway via the trans-Golgi network of the Golgi apparatus (an endosomal pathway) as well as autophagosome formation (an autophagic process) in skeletal muscles. Vacuoles may subsequently accumulate secondary to abnormal formation or turnover of autolysosomes at or after fusion of autophagosomes with early endosomes.     

Isolated third and fourth ventricles associated with neurosarcoidosis successfully treated by neuroendoscopy--case report

A 48-year-old woman presented with a unique case of isolated third and fourth ventricles associated with neurosarcoidosis, manifesting as upward gaze palsy. She had a past history of surgery for intracranial sarcoid granuloma 10 years previously and placement of a ventriculoperitoneal shunt 3 years later. Magnetic resonance imaging revealed isolated third and fourth ventricles. Endoscopic plasty of the right foramen of Monro, fenestration of the septum pellucidum, and revision of the ventriculoperitoneal shunt were performed. Postoperatively the size of the third and fourth ventricles was reduced, and the upward gaze palsy resolved. Endoscopic treatment is less invasive and effective for the treatment of multiple septum formation in the ventricular system.     

Load on the low back of care workers in nursing homes for the elderly]

In order to evaluate the load on the low back of care workers in nursing homes for the elderly, basic activities and working postures were analyzed for six care workers using the 30-s snap reading method. The trunk inclination angle (TIA) was also measured continuously using an inclination monitor. The analysis of basic activities showed that 22.5% and 21.1% of the workshift were spent on 'bathing and ablution' and 'meal', respectively, and 9.3%, 8.7% and 8.3% were spent on 'assistance with elimination', 'assistance with moving and repositioning' and 'exchange of bed sheet', respectively. Total of the assistance activity was 43.7% of the workshift. The analysis of working posture showed that 'standing' and 'standing bent forward' accounted for 36.1% and 29.5%, respectively, of the workshift. Total of three postures loading the low back ('standing bent forward', 'squatting', 'kneeling') accounted for 39.0%. The time spent in TIA of 20 degrees or more was 45.7%. The postures loading the low back during 'bathing and ablution', 'exchange of bed sheet' and 'assistance with elimination' account for 68.3%, 58.2% and 49.6%, respectively, which suggests that these activities load the low back of the care workers considerably.     

Studies on efficient methods for detection of Legionella species from environmental water by use of its selective media

To examine the optimal pH range for growth on media, growth of Legionella spp. on its selective media, BCYE alpha, WYO alpha and MWY agar media, in a pH range of 6.0-8.0 (at 0.5 intervals) was compared. The growth of two strains of L. pneumophila and one strain of L. micdadei on a WYO alpha agar supplemented with some selecting antimicrobial agents was markedly inhibited at all pH range except 6.0 and 7.0, suggesting a narrow optimal pH range for growth of these species compared to the BCYE alpha without selecting antimicrobioal agents. Vancomycin (VCM) added to the selective agar suppressed the growth of some Legionella spp. depending on the concentration. However, the extent of suppression was different among species and/or strains of Legionella spp. The selectivity for species other than Legionella spp. was also affected similarly by VCM concentration added to their media, suggesting that it is important to use proper amounts of the selecting antimicrobial agent depending on the species and/or strains of Legionella spp. or the other species in water samples. Amphothericin B (AMPH-B) added to a selective medium, MWY agar, in the concentration of 80 micrograms/ml hardly affected the growth of Legionella spp. examined, but effectively inhibited the growth of fungal strains identified as Aspergillus sp., Trichoderma sp., Scolecobasidium sp. and Mucor sp. which were isolated from cooling-tower water samples together with Legionella spp. Furthermore, the growth of a combination culture of one each of the 4 strains of isolated fungi and one each of the 3 strains of Legionella spp. was examined at various concentration of AMPH-B. Addition of AMPH-B to the selective medium at the concentration of 80 micrograms/ml suppressed the growth of spreading fungi, permitting the growth of Legionella spp. to allow efficient detection of the species.     

Temporal lobe epilepsy and corpora amylacea in the hippocampus: clinicopathologic correlation

Corpora amylacea (CoA) have been found in about 60% of neurosurgical specimens showing hippocampal sclerosis (HS). To determine clinical and neuroimaging differences between HS with and without CoA, we studied 29 patients (21 male, 8 female; age at surgery, 12 to 49 years) who underwent anterior temporal lobectomy for intractable medial temporal lobe epilepsy. No CoA were noted in the hippocampus of 11 cases, and deposition of CoA was mild and limited to the subependymal and vestigial hippocampal sulcus regions in nine cases; in nine cases, moderate to marked deposition was noted in the pyramidal cell layer, accompanying severe neuronal loss. No significant differences were evident between these three groups for age at onset, frequency and duration of epileptic seizures, the average age at surgery, or surgical results. Hippocampal hyperintensity in fluid-attenuated inversion recovery magnetic resonance images tended to increase with increasing hippocampal deposition of CoA. Formation of CoA appears to be a response to neuronal loss in the pyramidal cell layer, being related to the epileptogenic process as a consequence rather than a cause.     

Cyclic phosphatidic acid inhibits RhoA-mediated autophosphorylation of FAK at Tyr-397 and subsequent tumor-cell invasion

We demonstrated previously that rat ascites hepatoma MM1 cells require both lysophosphatidic acid (LPA) and fibronectin (FN) for phagokinetic motility and transcellular migration and that these events are regulated through the RhoA-ROCK pathway and tyrosine phosphorylation of proteins including focal adhesion kinase (FAK). Moreover, we reported that palmitoyl-cyclic phosphatidic acid (Pal-cPA), a structural analogue of LPA, inhibits LPA-induced migration of MM1 cells and experimental metastasis of B16 murine melanoma cells. However, the molecular mechanisms of action of Pal-cPA remains to be clarified. To examine this, total cellular lysates after stimulation with LPA or FN were subjected to time-course immunoblot analysis with anti-phophotyrosine and anti-pY397-FAK antibodies. Tyrosine-phosphorylation of FAK especially at Tyr-397 was obviously persistent after stimulation with LPA + FN compared to after stimulation with LPA alone. This persistent phosphorylation was necessary for MM1 cell migration and inhibited by Pal-cPA as by C3 exoenzyme Rho inhibitor. RhoA activity (GTP-bound RhoA) was also measured by the pull down assay using the Rho binding domain of Rhotekin. LPA-induced RhoA-activation of MM1 cells was completely inhibited by Pal-cPA. Moreover, we demonstrated that autophosphorylation of FAK at Tyr-397, downstream of RhoA, contributed to formation of focal adhesions and was critical in LPA-induced MM1 cell migration by developing autophosphorylation-deficient (Y397F) FAK-transfectants. Collectively, Pal-cPA hampered LPA-induced morphological changes and transcellular migration of MM1 cells through downregulating active RhoA and inhibiting its downstream events including autophosphorylation of FAK. Pal-cPA also inhibited endogenous (LPA-independent) activation of RhoA in human fibrosarcoma HT-1080 cells. Pal-cPA may potentially provide a new therapy for the treatment of cancer invasion and metastasis.