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1559 Establishment and biological characterization of an immortalized rat astrocyte line by SV40 large T antigen
$496
Friday, November 11, 2005
Zalnani, B 1 Mehrpoor, M :. llran University Of Medical Sciences, Iran Background: Stroke is more c o m m o n in patients with cerebral microembolism. Frequency of cerebral microembolism (high intensity transient siguals-HITS) in acute myocardial infarction was about 17"/o in one study. Streptokinase doesn't increase the risk of stroke following acute MI, we wanted to study the frequency of cerebral microembolism in acute M I patients who were treated with streptokinase. Methods: We have selected 65 patients with acute MI who were treated with streptokinase as a S K group and 65 patients with acute M I without thrombolytic therapy as a control group, during 72 hours of admission to coronary care unit. We used T C D to monitor for cerebral microembolism in both groups. All patients h a d 2-dimensional echocardiogram during study. Excluding criteria were prosthetic heart valves, carotid stenosis > 50"/o, poor window for T C D monitoring. Results: Mean age of SK group (158) was significantly lower than control group (65) p -- 0.003 but frequency of anterolateral MI, hypokinetic LV segments and ejection fraction were not significantly different between both groups. HITS were detected in 20% of SK group (mean 0.66 ± 0.99) a n d 10% of control group (mean 0.33 ± 0.71), so there is a significant increase of frequency of cerebral microembolism in S K group( p -- 0.034 ) Conclusion: HITS are detected in 30% of acute MI that were increased with streptokinase therapy although clinically there is no increment of stroke after thrombolytic therapy but maybe there is some difference in silent and asymptomatic stroke following thrombolytic therapy. 1558 Impact of low backache on health workers at Kenyatta National Hospital Anmyo, E 1, Mutiso, V 1, Kwasa, T 1. 1University of Nairobi, Kenya Background: Low back pain is a major problem that affects workers all over the world. It n o t only leads to absenteeism from but also loss in productivity. The economic, cost of the ailments is one of the highest of all medical conditions among workers. The aim of the study was to determine the prevalence of low back pain and its impact on workers at the Kenyatta National Hospital Method: A cross sectional survey of randomly selected health workers was performed using validated and pre-tested, self -administered close ended questionnaire. Results: 315(59°4) out of 527 workers recruited h a d low back ache . The m e a n age was 37.5 + 8 years with a range of 23 to 60.55.4 percent were females while the rest were males. Only 13"/o of the workers were able to perform their activities when they had backache while 3% required bed rest. 24 % of the worker h a d missed work in the last year due to backache compared to 12.7% due to other illness there being a statistical significant difference at a p value of 0.035. The m e a n duration of missed workdays was 15 +7 days. 90 °4 Of the workers h a d their social activities affected by the backache. Conclusion: There is a very high prevalence of low backache amongst hospital workers. The backache results in significant loss o f man-days and sodal aspects of the worker. 1559 Es~tablishment and biological characterization of an immortalized rat astrocyte line by SV40 large T antigen An, K 1. ~Department of Anesthesiology, Tongji Hospital, Tongji
Medical College, Wuhan, PR China Objective: To establish an immortalized rat astrocyte line and study its biological characterization for transgenic cellular implantation used for pain therapy. Methods: Rat cerebral cortical astrocytes were cultured according to the method of differential cell adhesiveness and then transfected with plasmJd p C M V S V 4 0 T / P U R which containing the simian virus 40 large
Poster Abstracts tumour antigen (SV40Tag) gene. The positive colonies were isolated by puromycin 1.5 ug/ml selection and expanded to many passages. P C R , R T - P C R and inmmnohistochenaJcal m e t h o d were used to detect the integtating and expression of large T antigen gene a n d its immunoreactivity of G F A P . Morphology feature of cells was observed by light and electron microscopy. Growth curve, culturing in soft agar, nude mice transplantation, immunhistochemistry and flow cytometry were used to investigate its biological characterization. Results: Cell colonies transformed with SV40Tag e D N A were isolated and subcultured, the SV40Tag gene has been transduced into their genome stably, the expression of SV40Tag gene in the cells was demonstrated in both the gene and protein level. A n immortalized rat astrocyte line(IAST)was established and maintained for more than 50 passages. The cells remained monolayer, anchorage dependent and attachinent-inhJbited growth. Subculture, freezing and recovering h a d no effect on cellular shape a n d proliferation of IAST. Proliferation capability of IAST was stronger than that of non-transformed cells, proliferation index a n d population doubling time were 29.17% and 19.58 h o u r respectively(P < 0.05). It could n o t form clone in soft agar and tumor in nude mice, but it still remained the same differentiated phenotypes as its original cells a n d inmmnostained positively for GFAP. Conclusion: Rat astrocytes can be immortalized by transferring exogenous SV40Tag gene, IAST immortalized with SV40Tag proliferates vigorously in vitro and is not belong to malignant transformed cells, so IAST could be used as cell carrier in transgenic cellular implantation for pain therapy.
1560 The role of Type A Botulinum Toxin in relieving pain in Cervical Dys~tonia Bertolasi, L ~, Bottanelli, M l, Vicentini, S ~. 1Department of Neurological
Sciences and Vision, Section of Clinical Neurology', University of Verona, Italy Cervical dystonia is the most c o m m o n form of the focal dystonias, and is characterized by an involuntary twisting and turning of the neck caused by abnormal involuntary sustained muscle contractions. Dystonic muscle activity can be mainly tonic, myoclonic, tremulous or a complex mixture. The clinical features of cervical dystonia are now well clarified. The vast majority of cases are idiopathic. Secondary cervical dystonia h a s been considered u n c o m m o n and could be associated with vascular, traumatic, infectious and toxic processes of the central a n d peripheral nervous systems. It could accompany degenerative disease such as progressive supranuclear palsy and cortical-basal ganglionic degeneration. A n important clinical feature in cervical dystonia is pain, often severe. T o d a y is well known that botulinum toxin is the most effective treatment for cervical dystonia. Several recent studies h a d demonstrated that type A botulinum toxin could block the release not only of acethilcoline from neuro-muscular junction but also of pain neu ro-mediators such as P substance and NO. In patients with cervical dystonia treated with botulinum toxin a relief of pain is usually observed, typically before than the muscular spasm is resolved, a n d lasts more. A possible analgesic role per s6 of botulinum toxin h a s been suggested. To test this hypothesis we administered a questionnaire (Neck Pain Questionnaire NPQ) to 15 consecutive patients treated with type A botulinmn toxin (Dyspor(rM) in our Department for cervical dystonia. 80 % of our patients experimented a complete relief of pain with the treatment. The remaining 20°4 showed a marked reduction of it. Our results confirm that pain is the first s y m p t o m to improve (1-2 days after the treatment), while sustained muscle contractions resolve later (range: 4-10 days after). Patients referee a long-lasting pain effect of botulinum toxin (range: 10-16 weeks), while involuntary postures appear after 8-12 weeks from the treatment. Our study suggests that botulinmn toxin could work in two ways, relieving pain and reducing muscular spasms.
Friday, November 11, 2005
Zalnani, B 1 Mehrpoor, M :. llran University Of Medical Sciences, Iran Background: Stroke is more c o m m o n in patients with cerebral microembolism. Frequency of cerebral microembolism (high intensity transient siguals-HITS) in acute myocardial infarction was about 17"/o in one study. Streptokinase doesn't increase the risk of stroke following acute MI, we wanted to study the frequency of cerebral microembolism in acute M I patients who were treated with streptokinase. Methods: We have selected 65 patients with acute MI who were treated with streptokinase as a S K group and 65 patients with acute M I without thrombolytic therapy as a control group, during 72 hours of admission to coronary care unit. We used T C D to monitor for cerebral microembolism in both groups. All patients h a d 2-dimensional echocardiogram during study. Excluding criteria were prosthetic heart valves, carotid stenosis > 50"/o, poor window for T C D monitoring. Results: Mean age of SK group (158) was significantly lower than control group (65) p -- 0.003 but frequency of anterolateral MI, hypokinetic LV segments and ejection fraction were not significantly different between both groups. HITS were detected in 20% of SK group (mean 0.66 ± 0.99) a n d 10% of control group (mean 0.33 ± 0.71), so there is a significant increase of frequency of cerebral microembolism in S K group( p -- 0.034 ) Conclusion: HITS are detected in 30% of acute MI that were increased with streptokinase therapy although clinically there is no increment of stroke after thrombolytic therapy but maybe there is some difference in silent and asymptomatic stroke following thrombolytic therapy. 1558 Impact of low backache on health workers at Kenyatta National Hospital Anmyo, E 1, Mutiso, V 1, Kwasa, T 1. 1University of Nairobi, Kenya Background: Low back pain is a major problem that affects workers all over the world. It n o t only leads to absenteeism from but also loss in productivity. The economic, cost of the ailments is one of the highest of all medical conditions among workers. The aim of the study was to determine the prevalence of low back pain and its impact on workers at the Kenyatta National Hospital Method: A cross sectional survey of randomly selected health workers was performed using validated and pre-tested, self -administered close ended questionnaire. Results: 315(59°4) out of 527 workers recruited h a d low back ache . The m e a n age was 37.5 + 8 years with a range of 23 to 60.55.4 percent were females while the rest were males. Only 13"/o of the workers were able to perform their activities when they had backache while 3% required bed rest. 24 % of the worker h a d missed work in the last year due to backache compared to 12.7% due to other illness there being a statistical significant difference at a p value of 0.035. The m e a n duration of missed workdays was 15 +7 days. 90 °4 Of the workers h a d their social activities affected by the backache. Conclusion: There is a very high prevalence of low backache amongst hospital workers. The backache results in significant loss o f man-days and sodal aspects of the worker. 1559 Es~tablishment and biological characterization of an immortalized rat astrocyte line by SV40 large T antigen An, K 1. ~Department of Anesthesiology, Tongji Hospital, Tongji
Medical College, Wuhan, PR China Objective: To establish an immortalized rat astrocyte line and study its biological characterization for transgenic cellular implantation used for pain therapy. Methods: Rat cerebral cortical astrocytes were cultured according to the method of differential cell adhesiveness and then transfected with plasmJd p C M V S V 4 0 T / P U R which containing the simian virus 40 large
Poster Abstracts tumour antigen (SV40Tag) gene. The positive colonies were isolated by puromycin 1.5 ug/ml selection and expanded to many passages. P C R , R T - P C R and inmmnohistochenaJcal m e t h o d were used to detect the integtating and expression of large T antigen gene a n d its immunoreactivity of G F A P . Morphology feature of cells was observed by light and electron microscopy. Growth curve, culturing in soft agar, nude mice transplantation, immunhistochemistry and flow cytometry were used to investigate its biological characterization. Results: Cell colonies transformed with SV40Tag e D N A were isolated and subcultured, the SV40Tag gene has been transduced into their genome stably, the expression of SV40Tag gene in the cells was demonstrated in both the gene and protein level. A n immortalized rat astrocyte line(IAST)was established and maintained for more than 50 passages. The cells remained monolayer, anchorage dependent and attachinent-inhJbited growth. Subculture, freezing and recovering h a d no effect on cellular shape a n d proliferation of IAST. Proliferation capability of IAST was stronger than that of non-transformed cells, proliferation index a n d population doubling time were 29.17% and 19.58 h o u r respectively(P < 0.05). It could n o t form clone in soft agar and tumor in nude mice, but it still remained the same differentiated phenotypes as its original cells a n d inmmnostained positively for GFAP. Conclusion: Rat astrocytes can be immortalized by transferring exogenous SV40Tag gene, IAST immortalized with SV40Tag proliferates vigorously in vitro and is not belong to malignant transformed cells, so IAST could be used as cell carrier in transgenic cellular implantation for pain therapy.
1560 The role of Type A Botulinum Toxin in relieving pain in Cervical Dys~tonia Bertolasi, L ~, Bottanelli, M l, Vicentini, S ~. 1Department of Neurological
Sciences and Vision, Section of Clinical Neurology', University of Verona, Italy Cervical dystonia is the most c o m m o n form of the focal dystonias, and is characterized by an involuntary twisting and turning of the neck caused by abnormal involuntary sustained muscle contractions. Dystonic muscle activity can be mainly tonic, myoclonic, tremulous or a complex mixture. The clinical features of cervical dystonia are now well clarified. The vast majority of cases are idiopathic. Secondary cervical dystonia h a s been considered u n c o m m o n and could be associated with vascular, traumatic, infectious and toxic processes of the central a n d peripheral nervous systems. It could accompany degenerative disease such as progressive supranuclear palsy and cortical-basal ganglionic degeneration. A n important clinical feature in cervical dystonia is pain, often severe. T o d a y is well known that botulinum toxin is the most effective treatment for cervical dystonia. Several recent studies h a d demonstrated that type A botulinum toxin could block the release not only of acethilcoline from neuro-muscular junction but also of pain neu ro-mediators such as P substance and NO. In patients with cervical dystonia treated with botulinum toxin a relief of pain is usually observed, typically before than the muscular spasm is resolved, a n d lasts more. A possible analgesic role per s6 of botulinum toxin h a s been suggested. To test this hypothesis we administered a questionnaire (Neck Pain Questionnaire NPQ) to 15 consecutive patients treated with type A botulinmn toxin (Dyspor(rM) in our Department for cervical dystonia. 80 % of our patients experimented a complete relief of pain with the treatment. The remaining 20°4 showed a marked reduction of it. Our results confirm that pain is the first s y m p t o m to improve (1-2 days after the treatment), while sustained muscle contractions resolve later (range: 4-10 days after). Patients referee a long-lasting pain effect of botulinum toxin (range: 10-16 weeks), while involuntary postures appear after 8-12 weeks from the treatment. Our study suggests that botulinmn toxin could work in two ways, relieving pain and reducing muscular spasms.