Peripheral nerve injuries result in the rapid degeneration of distal nerve segments and immediate loss of motor and sensory functions;behavioral recovery is typically poor.We used a plasmalemmal fusogen,polyethylene g...Peripheral nerve injuries result in the rapid degeneration of distal nerve segments and immediate loss of motor and sensory functions;behavioral recovery is typically poor.We used a plasmalemmal fusogen,polyethylene glycol(PEG),to immediately fuse closely apposed open ends of severed proximal and distal axons in rat sciatic nerves.We have previously reported that sciatic nerve axons repaired by PEG-fusion do not undergo Wallerian degeneration,and PEG-fused animals exhibit rapid(within 2–6 weeks)and extensive locomotor recovery.Furthermore,our previous report showed that PEG-fusion of severed sciatic motor axons was non-specific,i.e.,spinal motoneurons in PEG-fused animals were found to project to appropriate as well as inappropriate target muscles.In this study,we examined the consequences of PEG-fusion for sensory axons of the sciatic nerve.Young adult male and female rats(Sprague–Dawley)received either a unilateral single cut or ablation injury to the sciatic nerve and subsequent repair with or without(Negative Control)the application of PEG.Compound action potentials recorded immediately after PEG-fusion repair confirmed conduction across the injury site.The success of PEG-fusion was confirmed through Sciatic Functional Index testing with PEG-fused animals showing improvement in locomotor function beginning at 35 days postoperatively.At 2–42 days postoperatively,we anterogradely labeled sensory afferents from the dorsal aspect of the hindpaw following bilateral intradermal injection of wheat germ agglutinin conjugated horseradish peroxidase.PEG-fusion repair reestablished axonal continuity.Compared to unoperated animals,labeled sensory afferents ipsilateral to the injury in PEG-fused animals were found in the appropriate area of the dorsal horn,as well as inappropriate mediolateral and rostrocaudal areas.Unexpectedly,despite having intact peripheral nerves,similar reorganizations of labeled sensory afferents were also observed contralateral to the injury and repair.This central reorganization may contribute to the improved behavioral recovery seen after PEG-fusion repair,supporting the use of this novel repair methodology over currently available treatments.展开更多
Behavioral recovery using(viable)peripheral nerve allografts to repair ablation-type(segmental-loss)peripheral nerve injuries is delayed or poor due to slow and inaccurate axonal regeneration.Furthermore,such peripher...Behavioral recovery using(viable)peripheral nerve allografts to repair ablation-type(segmental-loss)peripheral nerve injuries is delayed or poor due to slow and inaccurate axonal regeneration.Furthermore,such peripheral nerve allografts undergo immunological rejection by the host immune system.In contrast,peripheral nerve injuries repaired by polyethylene glycol fusion of peripheral nerve allografts exhibit excellent behavioral recovery within weeks,reduced immune responses,and many axons do not undergo Wallerian degeneration.The relative contribution of neurorrhaphy and polyethylene glycol-fusion of axons versus the effects of polyethylene glycol per se was unknown prior to this study.We hypothesized that polyethylene glycol might have some immune-protective effects,but polyethylene glycol-fusion was necessary to prevent Wallerian degeneration and functional/behavioral recovery.We examined how polyethylene glycol solutions per se affect functional and behavioral recovery and peripheral nerve allograft morphological and immunological responses in the absence of polyethylene glycol-induced axonal fusion.Ablation-type sciatic nerve injuries in outbred Sprague–Dawley rats were repaired according to a modified protocol using the same solutions as polyethylene glycol-fused peripheral nerve allografts,but peripheral nerve allografts were loose-sutured(loose-sutured polyethylene glycol)with an intentional gap of 1–2 mm to prevent fusion by polyethylene glycol of peripheral nerve allograft axons with host axons.Similar to negative control peripheral nerve allografts not treated by polyethylene glycol and in contrast to polyethylene glycol-fused peripheral nerve allografts,animals with loose-sutured polyethylene glycol peripheral nerve allografts exhibited Wallerian degeneration for all axons and myelin degeneration by 7 days postoperatively and did not recover sciatic-mediated behavioral functions by 42 days postoperatively.Other morphological signs of rejection,such as collapsed Schwann cell basal lamina tubes,were absent in polyethylene glycol-fused peripheral nerve allografts but commonly observed in negative control and loose-sutured polyethylene glycol peripheral nerve allografts at 21 days postoperatively.Loose-sutured polyethylene glycol peripheral nerve allografts had more pro-inflammatory and less anti-inflammatory macrophages than negative control peripheral nerve allografts.While T cell counts were similarly high in loose-sutured-polyethylene glycol and negative control peripheral nerve allografts,loose-sutured polyethylene glycol peripheral nerve allografts expressed some cytokines/chemokines important for T cell activation at much lower levels at 14 days postoperatively.MHCI expression was elevated in loose-sutured polyethylene glycol peripheral nerve allografts,but MHCII expression was modestly lower compared to negative control at 21 days postoperatively.We conclude that,while polyethylene glycol per se reduces some immune responses of peripheral nerve allografts,successful polyethylene glycol-fusion repair of some axons is necessary to prevent Wallerian degeneration of those axons and immune rejection of peripheral nerve allografts,and produce recovery of sensory/motor functions and voluntary behaviors.Translation of polyethylene glycol-fusion technologies would produce a paradigm shift from the current clinical practice of waiting days to months to repair ablation peripheral nerve injuries.展开更多
To study the effect of free cells (suspended bacteria) on performance of entrapped bacteria system (i.e. polyethylene glycol (PEG)-pellet reactor) to treat NH4-N contaminated groundwater, two PEG-pellet reactors with ...To study the effect of free cells (suspended bacteria) on performance of entrapped bacteria system (i.e. polyethylene glycol (PEG)-pellet reactor) to treat NH4-N contaminated groundwater, two PEG-pellet reactors with a lot of free cells - Reactor A containing PEG-pellet and Reactor B containing PEG-pellet and supporting material - and the another control reactor without free cells (Reactor C) were set-up. Three reactors were operated under various NH4-N concentrations (40-60 mg/L) and various temperatures (5-25oC). The results show that the free cells effected on the NH4-N removal efficiency significantly. The free cells developed to be a biofilm layer on the pellet surface for Reactor A, the biofilm layer caused the decreasing NH4-N diffusion and incomplete nitrification eventually. On the other hand, most free cells attached to the supporting material for Reactor B. Although the NH4-N could diffuse properly, the free cells consuming acetate caused the added acetate was insufficient for complete denitrification. However, the results suggest that the supporting material could reduce the effect of free cells on the reactor performance at low temperature as indicated by 1) higher efficiency and 2) lower activation energy (Ea) for nitrification and denitrification in Reactor B than Reactor A.展开更多
This study was carried out to investigate the effects of silicon (Si) 3.6 mM (as calcium silicate) under drought stress induced by polyethylene glycol “PEG” at 15% (MW 8000), in addition to the control treatment on ...This study was carried out to investigate the effects of silicon (Si) 3.6 mM (as calcium silicate) under drought stress induced by polyethylene glycol “PEG” at 15% (MW 8000), in addition to the control treatment on growth and some biochemical constituents of date palm cv. Barhee cultured in vitro. Drought stress (15% PEG) depressed the growth of shoot and decreased protein content and chlorophyll concentration. Addition of 3.6 mM Si could improve the growth of shoot and increase the protein content and leaf chlorophyll concentrations of stressed plants. The inclusion of Si to the PEG containing medium significantly increased the catalase (CAT) and superoxide dismutase (SOD) activity in regenerated shoot, compared to other treatments. As well as drought stress 15% PEG induced significant accumulation of shoots proline, which were decreased by added silicon. Moreover, the results were also supported by the observation that PEG stress-induced decrease the response percentage of root induction and root lengths was reversed by added silicon. Addition of Si obviously significantly increased the wax content in leaves, response percentage of root induction and root lengths of plantlets under drought stress. The results of this study indicate that the application of silicon improved growth attributes, effectively mitigate the adverse effect of drought, and increase tolerance of date palm plants for drought stress during the course of date palm tissue cultures.展开更多
基金supported by the Department of Defense AFIRMⅢW81XWH-20-2-0029 grant subcontractLone Star Paralysis gift,UT POC19-1774-13 grant+1 种基金Neuraptive Therapeutics Inc.26-7724-56 grantNational Institutes of Health R01-NS128086(all to GDB)。
文摘Peripheral nerve injuries result in the rapid degeneration of distal nerve segments and immediate loss of motor and sensory functions;behavioral recovery is typically poor.We used a plasmalemmal fusogen,polyethylene glycol(PEG),to immediately fuse closely apposed open ends of severed proximal and distal axons in rat sciatic nerves.We have previously reported that sciatic nerve axons repaired by PEG-fusion do not undergo Wallerian degeneration,and PEG-fused animals exhibit rapid(within 2–6 weeks)and extensive locomotor recovery.Furthermore,our previous report showed that PEG-fusion of severed sciatic motor axons was non-specific,i.e.,spinal motoneurons in PEG-fused animals were found to project to appropriate as well as inappropriate target muscles.In this study,we examined the consequences of PEG-fusion for sensory axons of the sciatic nerve.Young adult male and female rats(Sprague–Dawley)received either a unilateral single cut or ablation injury to the sciatic nerve and subsequent repair with or without(Negative Control)the application of PEG.Compound action potentials recorded immediately after PEG-fusion repair confirmed conduction across the injury site.The success of PEG-fusion was confirmed through Sciatic Functional Index testing with PEG-fused animals showing improvement in locomotor function beginning at 35 days postoperatively.At 2–42 days postoperatively,we anterogradely labeled sensory afferents from the dorsal aspect of the hindpaw following bilateral intradermal injection of wheat germ agglutinin conjugated horseradish peroxidase.PEG-fusion repair reestablished axonal continuity.Compared to unoperated animals,labeled sensory afferents ipsilateral to the injury in PEG-fused animals were found in the appropriate area of the dorsal horn,as well as inappropriate mediolateral and rostrocaudal areas.Unexpectedly,despite having intact peripheral nerves,similar reorganizations of labeled sensory afferents were also observed contralateral to the injury and repair.This central reorganization may contribute to the improved behavioral recovery seen after PEG-fusion repair,supporting the use of this novel repair methodology over currently available treatments.
基金supported by grants from the Lone Star Paralysis Foundation,NIH R01NS081063Department of Defense award W81XWH-19-2-0054 to GDB+2 种基金supported by University of Wyoming Startup funds,Department of Defense grant W81XWH-17-1-0402the University of Wyoming Sensory Biology COBRE under National Institutes of Health(NIH)award number 5P20GM121310-02the National Institute of General Medical Sciences of the NIH under award number P20GM103432 to JSB。
文摘Behavioral recovery using(viable)peripheral nerve allografts to repair ablation-type(segmental-loss)peripheral nerve injuries is delayed or poor due to slow and inaccurate axonal regeneration.Furthermore,such peripheral nerve allografts undergo immunological rejection by the host immune system.In contrast,peripheral nerve injuries repaired by polyethylene glycol fusion of peripheral nerve allografts exhibit excellent behavioral recovery within weeks,reduced immune responses,and many axons do not undergo Wallerian degeneration.The relative contribution of neurorrhaphy and polyethylene glycol-fusion of axons versus the effects of polyethylene glycol per se was unknown prior to this study.We hypothesized that polyethylene glycol might have some immune-protective effects,but polyethylene glycol-fusion was necessary to prevent Wallerian degeneration and functional/behavioral recovery.We examined how polyethylene glycol solutions per se affect functional and behavioral recovery and peripheral nerve allograft morphological and immunological responses in the absence of polyethylene glycol-induced axonal fusion.Ablation-type sciatic nerve injuries in outbred Sprague–Dawley rats were repaired according to a modified protocol using the same solutions as polyethylene glycol-fused peripheral nerve allografts,but peripheral nerve allografts were loose-sutured(loose-sutured polyethylene glycol)with an intentional gap of 1–2 mm to prevent fusion by polyethylene glycol of peripheral nerve allograft axons with host axons.Similar to negative control peripheral nerve allografts not treated by polyethylene glycol and in contrast to polyethylene glycol-fused peripheral nerve allografts,animals with loose-sutured polyethylene glycol peripheral nerve allografts exhibited Wallerian degeneration for all axons and myelin degeneration by 7 days postoperatively and did not recover sciatic-mediated behavioral functions by 42 days postoperatively.Other morphological signs of rejection,such as collapsed Schwann cell basal lamina tubes,were absent in polyethylene glycol-fused peripheral nerve allografts but commonly observed in negative control and loose-sutured polyethylene glycol peripheral nerve allografts at 21 days postoperatively.Loose-sutured polyethylene glycol peripheral nerve allografts had more pro-inflammatory and less anti-inflammatory macrophages than negative control peripheral nerve allografts.While T cell counts were similarly high in loose-sutured-polyethylene glycol and negative control peripheral nerve allografts,loose-sutured polyethylene glycol peripheral nerve allografts expressed some cytokines/chemokines important for T cell activation at much lower levels at 14 days postoperatively.MHCI expression was elevated in loose-sutured polyethylene glycol peripheral nerve allografts,but MHCII expression was modestly lower compared to negative control at 21 days postoperatively.We conclude that,while polyethylene glycol per se reduces some immune responses of peripheral nerve allografts,successful polyethylene glycol-fusion repair of some axons is necessary to prevent Wallerian degeneration of those axons and immune rejection of peripheral nerve allografts,and produce recovery of sensory/motor functions and voluntary behaviors.Translation of polyethylene glycol-fusion technologies would produce a paradigm shift from the current clinical practice of waiting days to months to repair ablation peripheral nerve injuries.
文摘To study the effect of free cells (suspended bacteria) on performance of entrapped bacteria system (i.e. polyethylene glycol (PEG)-pellet reactor) to treat NH4-N contaminated groundwater, two PEG-pellet reactors with a lot of free cells - Reactor A containing PEG-pellet and Reactor B containing PEG-pellet and supporting material - and the another control reactor without free cells (Reactor C) were set-up. Three reactors were operated under various NH4-N concentrations (40-60 mg/L) and various temperatures (5-25oC). The results show that the free cells effected on the NH4-N removal efficiency significantly. The free cells developed to be a biofilm layer on the pellet surface for Reactor A, the biofilm layer caused the decreasing NH4-N diffusion and incomplete nitrification eventually. On the other hand, most free cells attached to the supporting material for Reactor B. Although the NH4-N could diffuse properly, the free cells consuming acetate caused the added acetate was insufficient for complete denitrification. However, the results suggest that the supporting material could reduce the effect of free cells on the reactor performance at low temperature as indicated by 1) higher efficiency and 2) lower activation energy (Ea) for nitrification and denitrification in Reactor B than Reactor A.
文摘This study was carried out to investigate the effects of silicon (Si) 3.6 mM (as calcium silicate) under drought stress induced by polyethylene glycol “PEG” at 15% (MW 8000), in addition to the control treatment on growth and some biochemical constituents of date palm cv. Barhee cultured in vitro. Drought stress (15% PEG) depressed the growth of shoot and decreased protein content and chlorophyll concentration. Addition of 3.6 mM Si could improve the growth of shoot and increase the protein content and leaf chlorophyll concentrations of stressed plants. The inclusion of Si to the PEG containing medium significantly increased the catalase (CAT) and superoxide dismutase (SOD) activity in regenerated shoot, compared to other treatments. As well as drought stress 15% PEG induced significant accumulation of shoots proline, which were decreased by added silicon. Moreover, the results were also supported by the observation that PEG stress-induced decrease the response percentage of root induction and root lengths was reversed by added silicon. Addition of Si obviously significantly increased the wax content in leaves, response percentage of root induction and root lengths of plantlets under drought stress. The results of this study indicate that the application of silicon improved growth attributes, effectively mitigate the adverse effect of drought, and increase tolerance of date palm plants for drought stress during the course of date palm tissue cultures.
