Activation of migration of endogenous stem cells by erythropoietin as potential rescue for neurodegenerative diseases.

UNLABELLED: Neurodegenerative disorders such as Alzheimer's disease (AD) are characterized by progressive cognitive dysfunction and memory loss. There is deposition of amyloid plaques in the brain and subsequent neuronal loss. Neuroinflammation plays a key role in the pathogenesis of AD. There is still no effective curative therapy for these patients. One promising strategy involves the stimulation of endogenous stem cells. This study investigated the therapeutic effect of erythropoietin (EPO) in neurogenesis, and proved its manipulation of the endogenous mesenchymal stem cells in model of lipopolysaccharide (LPS)-induced neuroinflammation. METHODS: Forty five adult male mice were divided equally into 3 groups: Group I (control), group II (LPS untreated group): mice were injected with single dose of lipopolysaccharide (LPS) 0.8 mg/kg intraperitoneally (ip) to induce neuroinflammation, group III (EPO treated group): in addition to (LPS) mice were further injected with EPO in dose of 40 µg/kg of body weight three times weekly for 5 consecutive weeks. Groups were tested for their locomotor activity and memory using open field test and Y-maze. Cerebral specimens were subjected to histological and morphometric studies. Glial fibrillary acidic protein (GFAP) and mesenchymal stem cell marker CD44 were assessed using immunostaining. Gene expression of brain derived neurotrophic factor (BDNF) was examined in brain tissue. RESULTS: LPS decreased locomotor activity and percentage of correct choices in Y-maze test. Cerebral sections of LPS treated mice showed increased percentage area of dark nuclei and amyloid plaques. Multiple GFAP positive astrocytes were detected in affected cerebral sections. In addition, decrease BDNF gene expression was noted. On the other hand, EPO treated group, showed improvement in locomotor and cognitive function. Examination of the cerebral sections showed multiple neurons exhibiting less dark nuclei and less amyloid plaques in comparison to the untreated group. GFAP positive astrocytes were also reduced. Cerebral sections of the EPO treated group showed multiple branched and spindle CD44 positive cells inside and around blood vessels more than in LPS group. This immunostaining was negative in the control group. EPO administration increased BDNF gene expression. CONCLUSION: This study proved that EPO provides excellent neuroprotective and neurotrophic effects in vivo model of LPS induced neuroinflammation. It enhances brain tissue regeneration via stimulation of endogenous mesenchymal stem cells proliferation and their migration to the site of inflammation. EPO also up regulates cerebral BDNF expression and production, which might contributes to EPO mediated neurogenesis. It also attenuates reactive gliosis thus reduces neuroinflammation. These encouraging results obtained with the use of EPO proved that it may be a promising candidate for future clinical application and treatment of neurodegenerative diseases.

Involuntary action of the external anal sphincter. Manometric and electromyographic studies.

The changes in the rectal neck (anal canal) pressure and electromyography (EMG) of the external anal sphincter and levator ani muscle were studied in 20 dogs before and after internal anal sphincter excision by 2 weeks and monthly up to 10 months. The rectal neck pressure dropped to 41% of the preoperative level. It was then gradually elevated from the beginning of the 1st month following excision till it reached 88% of the level before excision at the 10th month. The external sphincter EMG basal activity after internal sphincterectomy by 2 weeks showed a decrease in the amplitude of the slow and high potential waves. From the 1st to the 7th month after excision, changes in the frequency and amplitude of the waves occurred, and by the 10th month, the external sphincter showed the characteristics of the EMG of both the internal and external anal sphincters. The basal activity of the levator ani muscle increased at the 10th month. As reported previously, the changes in the rectal neck pressure and in the EMG of the external anal sphincter after internal sphincterectomy seem to be due to the histological changes in the muscle and include striated and smooth muscle fiber hypertrophy and proliferation. These changes adapt the external sphincter to serve the function of not only voluntary but also involuntary continence.

A new concept of the anatomy of the anal sphincter mechanism and the physiology of defecation. The involuntary action of the external anal sphincter: histologic study.

The histologic changes in the external anal sphincter after internal anal sphincter excision were studied in 20 dogs. An external sphincter biopsy was taken before internal sphincterectomy and 2 weeks and monthly thereafter for 10 months. The excised material was studied microscopically after being stained with hematoxylin and eosin, Verhoeff-van Gieson and succinic dehydrogenase. 70% of external sphincter specimens before internal sphincter excision showed smooth muscle fibers scattered between the striated fibers. These smooth fibers could be responsible for the resting tone of the external sphincter. After internal sphincter excision, characteristic histologic changes could be identified in the external sphincter. From the 2nd week to the 5th month after excision, the external sphincter showed degenerative and hypertrophic changes. From the 6th to the 10th month, there were regeneration of the striated muscle fibers and increase in the number of smooth fibers so that by the 10th month a 'compound' muscle of striated and smooth fibers was identified. Two theories were put forward to explain the smooth fiber preponderance in the external sphincter after internal sphincter excision: mutant and replacement theories. The increased nonstriated element in the external sphincter seems to be a structural-functional adaptation so that the external sphincter takes on the involuntary function of the excised muscle.