Class of 2012 Abstract / Specific Aims: The goal of this project was to evaluate the use of a preparation of the brainstem and spinal cord of neonatal rats that has been widely used for observing and quantifying central nervous activity, as well as the response to pharmacological manipulation. To achieve this, we specifically aimed to remove the intact brainstem and spinal cord of newborn rats, and develop a preparation that would maintain physiological function and allow for recording of electrical activity. Methods: Multiple dissections were performed on neonatal rats. Conditions during the dissections were controlled to maintain physiological function. Once removed, the intact brainstem and spinal cord was placed in a preparation that allowed for manipulation and access to nerve rootlets. Finally, glass suction electrodes were used to record electrical activity directly from the nerve rootlets. Once recorded, the data were stored on a hard drive for further analysis. Main Results: We were successful in isolating the intact brainstem and spinal cord in neonatal rats while maintaining physiological conditions and nervous activity. The preparation allowed for easy access to nerve roots as well as customization for different experiments. We were also successful in recording nerve activity in the preparation and collection of data for use in future experiments Conclusions: We conclude that the brainstem spinal cord preparation described in this study is a valuable tool that allows for recording and analysis of nerve activity, and specifically for measurement of respiratory motor output. This is a preparation that can be used in a variety of experiments that attempt to observe or quantify the activity of central nerve cells and allows for pharmacological interventions that could be applied in various experiments.

M.Tech. / Purpose: To compare the efficacy of diversified side posture adjustments and segmental drop piece adjustments in the treatment of sacroiliac joint dysfunction. Method: Thirty-two participants were randomly divided in two equal groups. Group A (n = 16) received diversified side posture adjustments and Group B (n = 16) received segmental drop piece adjustments. The trial consisted of seven sessions over a period of three weeks, of which the first six were treatment sessions, with the final seventh session serving the purpose of obtaining final post-treatment data. Data was obtained only at the first, third, fifth and seventh sessions. Objective data consisted of measuring lumbar spine range of motion using a digital inclinometer and pain pressure tolerance over the middle of the sacroiliac joint line by using a pressure algometer. Subjective data was obtained by having each participant complete a numerical pain rating scale and the Oswestry back pain and disability questionnaire.

M.Tech. / Purpose: This study aims to compare the effects of a manual high velocity, low amplitude spinal manipulation and a mechanical force, manually assisted spinal manipulation in the treatment of sub acute lumbar facet syndrome with regards to pain, disability, lumbar spine range of motion and pressure pain threshold. These effects were based on two questionnaires, the McGill Pain Questionnaire and the Oswestry Low Back Pain and Disability Questionnaire, lumbar spine range of motion readings using a digital inclinometer and pressure pain threshold readings taken with an algometer. Method: Thirty participants who met the inclusion criteria were randomly allocated to one of two groups. One group received a manual lumbar spine manipulation using the Diversified Technique. The second group received lumbar spine manipulative therapy with the use of the Activator Adjusting Instrument. Treatment occurred over a three week period, with participants receiving treatment six times out of a total of seven consultations. Procedure: Subjective data was collected using both the McGill Pain Questionnaire and the Oswestry Low Back Pain and Disability Questionnaire. Objective data was collected using a digital inclinometer to measure lumbar spine range of motion, and an algometer to measure pressure pain threshold. The questionnaires and all readings were recorded prior to treatment on the first, fourth and seventh visits. The data was then analysed by a statistician. Results: Clinical improvements in both groups were noted over the duration of the study with regards to pain, disability, left rotation and left and right lateral flexion. The group receiving a spinal manipulation with the Activator Adjusting Instrument also showed clinical improvement for flexion and right rotation and only the manual manipulation group improved in algometer measurements. Both groups displayed a decrease in lumbar extension readings over time. Statistical significance was observed for subjective measurements only. Conclusion: The results show that both spinal manipulative therapies are effective in the treatment of sub acute lumbar facet syndrome, however neither proved to be preferential over the other for subjective and objective measurements.

M.Tech. (Chiropractic) / Mechanical neck pain is the general term that refers to any type of pain within the cervical spine caused by placing abnormal stress and strain on muscles of the vertebral column. This is a very common musculo-skeletal disorder within the population, with 45% to 54% reporting neck pain at any given time of their lives (Martinez-Segura, Fernandez-de-las-Penas, Ruiz-Saez, Lopez-Jimenez and Rodriguez-Blanco, 2006). A vast majority of individuals do not experience a complete resolution of symptoms and as such often becomes a chronic pain (Cote, Cassidy, Carol and Kristman, 2004). Chronic pain by definition is one which is present for a period longer than six weeks (Segen, 2002). Accompanying the neck pain is often a limited or reduced range of motion within the cervical spine. Some studies suggest that hyper-tonicity and strain of the supporting muscles within this region due to an altered biomechanics also contributes to the pain and dysfunction (Armstrong, McNair and Williams, 2005 and Dall’Alba et al. 2001). One of the major muscle groups involved in this is the posterior cervical musculature. This is a general term used to describe the muscles located on the posterior aspect of the neck and includes longissimus capitis, semispinalis capitis, semispinalis cervicis, rotatores and multifidus (Dalley and Moore, 2006). Chiropractic adjustment therapy has been shown to have many effects of the body and especially the cervical spine including a decrease in pain perception and muscle hypertonicity (Kirkaldy-Willis and Cassidy, 1985), an increase in cervical range of motion (Bergmann, Peterson and Lawrence, 1993) as well as an increased skin pain tolerance level (Terrett and Vernon, 1984). Dry needling therapy has also been proven to have similar effects, including, a reduction in pain perception, an increase in range in motion of the specific linked biomechanical section, and most importantly a reduction of muscle tension (Travell and Simons, 1993). The purpose of this study was to see if a synergistic effect of the two above mentioned treatments would occur when combined under one treatment protocol. The aim of this study was to determine how effectively cervical adjustment therapy, dry needling of the posterior cervical muscles and a combination of the two treatments was in treating chronic mechanical neck pain with regards to pain, disability and cervical spine range of motion over a three week period. Participants were recruited from the University of Johannesburg Chiropractic Day Clinic. They were eligible to participate in the study once they met the inclusion and exclusion criteria. Participants were recruited by means of word of mouth as well as with the use of advertisements that were placed around the respective campuses of the University of Johannesburg. Thirty participants who presented with chronic mechanical neck pain, volunteered for this comparative study. This trial is a randomised controlled clinical trial which used convenience sampling. Group A received a combination treatment of dry needling and cervical manipulation. Group B received cervical adjustment only while group C received dry needling of the posterior cervical musculature alone. participants were treated for a total of 6 visits. Subjective and objective measurements were done at visits 1, 4 and a final visit 7 during which only measurements were taken. These effects are based on Vernon-Mior Pain and Disability Index, Numerical Pain Scale Rating, Pressure Algometer as well as Cervical Spine Range of Motion measuring instrument (CROM). The results of this trial indicated that all three treatments were effective in treating chronic mechanic neck pain. While one treatment was not statistically more effective than the other the dry needling and cervical manipulation alone produced a superior result. While combining these treatments was effective the synergistic effect one would expect was not as evident.

Dissertation submitted in partial compliance with the requirements for the Master's Diploma in Technology: Chiropractic at Technikon Natal, 1995. / The efficacy of chiropractic spinal manipulation on its own versus chiropractic spinal manipulation combined with other treatment modalities used in a chiropractic practice was evaluated in a single blind, randomised, controlled trial using a patient population of thirty nurses who were experiencing mechanical low back pain / M

Functional electrical stimulation (FES) techniques may be used to restore motor function lost or impaired through spinal cord injury. In order to use these techniques to restore complex tasks such as walking, it is necessary to provide sensory feedback to regulate the output of the FES controller. It has been suggested that multi-microelectrode probes (microprobes) implanted into the peripheral nervous system can be used to detect signals originating from the body's own sensors. These signals could be decoded and used to regulate the output of the FES controller. Prior to the present work, however, microprobes had primary been used to study neural activity in the brain, not peripheral nerves. In the present work, locust peripheral nerve has been used as an animal model for experimental and computer modelling work. The experimental work was directed at discerning the detail of information that can be obtained using microprobes to record from peripheral nerves (ie, the selectivity of the probes). In the computer modelling work, the effects of filtering the recorded signal were studied using an electrical circuit simulator programme (SPICE). Finite element analysis software (ANSYS) was used to model the electrical potential distribution in the nerve trunk, and to determine the effects of the probe substrate on the recorded signal. The results of the experimental work indicated that it may be possible to achieve higher selectivity in recording with microprobes than predicted by some models. It is concluded that future models need to represent the situation in greater detail in order to make more realistic predictions regarding the practical work. This will require further data on the electrical properties of the structures modelled within the nerve trunk. The SPICE modelling work successfully predicted the shape of the neural signals that would be recorded in the practical work. The partial differentiating effect of high pass filtering neural signals was also demonstrated. The results of the finite element modelling work demonstrated that the probe substrate would be expected to amplify signals from fibres directly in front of it, and attenuate signals from fibres behind it. This was shown to be significant for probe substrates with dimensions much smaller than the longitudinal spread of the action potential along the fibre. It was also found that these effects can be influenced by the position of the microprobe substrate relative to other structures within the nerve trunk; not just relative to the fibre. The significance of these results as they relate to mammalian nerve is discussed. Improved experimentation techniques and models are outlined, based on the results of this work. These include the requirement for improved facilities to determine the limits of selectivity in recording from peripheral nerves, and also the inclusion of inhomogeneities in models of the nerve trunk to make more realistic predictions regarding practical work. Finally, the development of active probes is discussed, including requirements for particularly novel circuitry, and the integration of many devices into a system to control FES.

Spinal Muscular Atrophy (SMA) is an autosomal, recessive form of childhood motor neuron disease and the most common genetic cause of infant mortality in the western world. SMA displays the characteristic hallmarks of a motor neuron disease, including loss of motor neurons in the spinal cord and atrophy of skeletal muscles. However, mounting evidence suggests that multiple tissues and body systems, beyond the neuromuscular system, are affected in SMA. Previous studies have highlighted alterations in the vascular system in both SMA patients and in a variety of mouse models of the disease, reporting alterations in vessel structure and perfusion abnormalities in peripheral tissues. In this project a detailed morphological investigation of the capillary beds of skeletal muscle and the spinal cord, two of the key pathological tissues in SMA was undertaken. This work was conducted in the Smn-/-;SMN2, Smn-/-;SMN2tg/+ and Smn-/-;SMN2;Δ7 mouse models of SMA. Significant alterations in the form and extent of the skeletal muscle and spinal cord capillary bed in SMA mice were identified, the most striking of which being a reduction in capillary density in SMA tissue when compared to control littermate tissue. In skeletal muscle, this reduction in capillary density was found to be a postnatal phenomenon, which occurred independently of denervation, in a variety of phenotypically distinct muscles and in all three SMA mouse models investigated. In the spinal cord, the capillary defect was seen to develop in a similar postnatal pattern to that observed in skeletal muscle. Importantly, a reduction in capillary density was observed in the ventral horn of the spinal cord, which houses motor neuron cell bodies, a known pathological target in SMA. These motor neurons were seen to be surrounded by fewer capillaries than their control counterparts. Using an injectable marker of hypoxia, it was determined that the cells of the ventral horn of SMA spinal cords are hypoxic. This suggests that the capillary defect identified has a functional impact on the tissues it is observed in. Having established the presence of capillary defect in SMA tissue, the effect of potential SMA therapeutics on the capillary defect was then investigated. The effect of HDAC inhibitors, which have been successfully shown to increase the levels of the disease causing Smn protein, was investigated. Treatment with the HDAC inhibitor SAHA was found to ameliorate the capillary defect, significantly improving capillary density in SMA skeletal muscle. This implies that the capillary defect is related to Smn levels in tissue and is amenable to therapeutics which increase Smn levels. Having characterised the capillary defect in SMA tissues in detail, a selection of tools were then used to investigate the underlying mechanisms resulting in the defect. First, using primary cell cultures, the growth and morphology of the key cellular component of capillaries, the endothelial cell, was examined. While displaying reduced levels of the Smn protein, endothelial cells isolated from SMA tissues showed no difference in growth rate, morphology or endothelial cell marker expression when compared to endothelial cells isolated from control tissue. This suggests that the defects seen in SMA capillary beds are not the result of defects in the structure and growth of endothelial cells. Second, retinas from SMA mice were found to exhibit similar capillary defects to those observed in SMA skeletal muscle and spinal cord. Given the entirely postnatal development of the retinal capillary network, the retina was identified as a useful experimental preparation for the further investigation of the mechanisms underlying the capillary defect in SMA. In summary, this work highlights the incidence and importance of capillary defects in mouse models of spinal muscular atrophy.

"A dissertation submitted in partial compliance with the requirements for the Master's Diploma in Technology: Chiropractic, Technikon Natal, 1995. / Mechanical lower back pain is a common clinical entity which encompasses a spectrum of disorders. studies show that spinal manipulative therapy is of value in the treatment of mechanical lower back pain, while some chiropractors are of the opinion that McManis traction offers an effective adjunct to spinal manipulative therapy in the treatment of such conditions / M

Abstract Genetic factors have been implicated to play a role in both degenerative lumbar spinal stenosis (LSS) and intervertebral disc disease (IDD). Sequence variations in the genes coding for collagen IX and inflammatory mediators have been indicated as risk factors for IDD. Nine genes coding for intervertebral disc (IVD) collagens I, II, IX and XI and aggrecan (AGC1) were analyzed for sequence variations in 29 Finnish individuals with LSS. In addition, two polymorphisms in the vitamin D receptor gene and one in the matrix metalloproteinase-3 gene were studied. Study subjects were analyzed both clinically and radiologically. Results indicated an association between the COL11A2 IVS6-4 a to t polymorphism and LSS (p = 0.0016). Moreover, the t/t genotype was found more often in the patient group compared to controls (p = 0.0011). A novel splicing mutation, likely resulting in the synthesis of a truncated protein, was identified in COL9A2. Eight hundred four Chinese individuals were screened for the presence of the Trp2 and Trp3 alleles. The Trp2 allele was found in 20% of the individuals compared to the previously reported 5% in Finnish patients with IDD characterized by sciatica. The Trp2 allele was found to predispose to IVD degeneration and end plate herniations, increasing the risk by 2.4-fold from 40 to 49 years of age. In addition, the degeneration was worse in individuals with the Trp2 allele. The risk for annular tears was 4-fold greater in study subjects from 30 to 39 years of age who were Trp2 positive. Surprisingly, the Trp3 allele was absent even though it was found in about 9% of Finnish individuals. One hundred fifty-five Finnish individuals with IDD characterized by sciatica were analyzed for sequence variations in four genes coding for inflammatory mediators IL1A, IL1B, IL6, and TNFA. In addition, sixteen polymorphisms in inflammatory mediator genes were analyzed. The results identified an association between sciatica and the E5+15T>A polymorphism in IL6 (p = 0.007). A significant association was also seen in the IL6 haplotype analysis (-597 g>a, -572 g>c, -174 g>c and E5+15T>A). The association of the GGGA haplotype with the disease was highly significant (p = 0.0033).

The childhood neuromuscular disease spinal muscular atrophy (SMA) is caused by low levels of survival motor neuron (SMN) protein. Historically, SMA has been characterised as a disease primarily affecting lower motor neurons. However, recent breakthroughs have revealed defects in other non-neuronal cells and tissues. In vivo analysis of peripheral nerve showed defects in Schwann cells, manifesting as abnormal myelination and delayed maturation of axo-glia interactions. The experiments in this thesis were designed to build on these observations and examine whether Schwann cell defects are intrinsic and occur as a primary result of low levels of SMN in that cell type, or rather represent a secondary consequence of pathology in neighbouring motor neurons. I initially developed a protocol to allow isolation of high-yields of purified, myelination-competent Schwann cells from ‘Taiwanese’ SMA mice. SMA-derived Schwann cells had significantly reduced SMN levels and failed to respond normally to differentiation cues. Increasing SMN levels restored myelin protein expression in Schwann cells from SMA mice. Perturbations in expression of key myelin proteins were likely due to failure of protein translation and/or stability rather than transcriptional defects. Co-cultures of healthy neurons with SMA Schwann cells revealed a significant reduction in myelination compared to cultures where wild-type Schwann cells were used. The presence of SMA Schwann cells also disrupted neurite stability. Perturbations in the expression of key extracellular matrix proteins, such as laminin α2, in SMA-derived Schwann cells suggests that Schwann cells were influencing neurite stability by modulating the composition of the extracellular matrix. Previous studies have demonstrated that low levels of SMN lead to disruption of ubiquitin homeostasis and decreased expression of ubiquitin-like modifier activating enzyme (UBA1) in the neuromuscular system, driving neuromuscular pathology via a beta-catenin dependent pathway. Label-free proteomics analysis of SMA and control Schwann cells identified 195 proteins with modified expression profiles. Bioinformatic analysis of these proteins using Ingenuity Pathway Analysis (IPA) software confirmed that major disruption of protein ubiquitination pathways was also present in Schwann cells from SMA mice. Immunolabeling and proteomics data both revealed that UBA1 levels were significantly reduced in SMA-derived Schwann cells. However, loss of UBA1 in Schwann cells did not lead to downstream modifications in beta-catenin pathways. Pharmacological inhibition of UBA1 in healthy Schwann cells was sufficient to induce defects in myelin protein expression, suggesting that UBA1 defects contribute directly to Schwann cell disruption in SMA. I conclude that low levels of SMN induce intrinsic defects in Schwann cells, mediated at least in part through disruption to ubiquitination pathways.