Mechanisms behind the Development of Chronic Low Back Pain and Its Neurodegenerative Features

Chronic back pain is complex and there is no guarantee that treating its potential causes will cause the pain to go away. Therefore, rather than attempting to “cure” chronic pain, many clinicians, caregivers and researchers aim to help educate patients about their pain and try to help them live a better quality of life despite their condition. A systematic review has demonstrated that patient education has a large effect on pain and pain related disability when done in conjunction with treatments. Therefore, understanding and updating our current state of knowledge of the pathophysiology of back pain is important in educating patients as well as guiding the development of novel therapeutics. Growing evidence suggests that back pain causes morphological changes in the central nervous system and that these changes have significant overlap with those seen in common neurodegenerative disorders. These similarities in mechanisms may explain the associations between chronic low back pain and cognitive decline and brain fog. The neurodegenerative underpinnings of chronic low back pain demonstrate a new layer of understanding for this condition, which may help inspire new strategies in pain education and management, as well as potentially improve current treatment

Engineering Cell Migration to Chemical and Physical Stimuli using Ca2+ Based Signalling

A major challenge of cell-based therapies is the appropriate and effective localization of cells to target sites. Engineering cell migration to a specific target site can offer a promising solution to this problem. Previously, a chimeric protein known as CaRQ was developed to modulate cell migration based on intracellular Ca2+ transients. CaRQ confers versatility in rewiring cell migration to chemical and physical stimuli, by combining CaRQ with receptors that generate Ca2+ transients. Through in vitro cell-based experiments, this thesis demonstrates the versatility of CaRQ in engineering cell migration in response to chemical (i.e. cytokines) and physical (i.e. magnetic fields) stimuli. First, migration towards vascular endothelial growth factor (VEGF) was engineered by a system composed of CaRQ and the VEGF receptor 2 (VEGFR2). VEGF binds to VEGFR2 and generates an intracellular Ca2+ transient. Stable cells expressing VEGFR2 and CaRQ displayed migratory blebbing, migration across porous membranes, directed migration, as well as wound healing in response to VEGF and VEGFR2 antibodies. Second, migration towards granulocyte-macrophage colony-stimulating factor (GM-CSF) was engineered by a system composed of CaRQ and a chimeric receptor (henceforth GMRchi). GMRchi is composed of the VEGFR2 cytoplasmic domain and GM-CSF receptor extracellular domain. Since the GM-CSF receptor alone cannot generate Ca2+transients, the chimeric receptor was engineered, demonstrating that migration is not limited to the repertoire of naturally occurring receptors. Stable cells expressing GMRchi and CaRQ displayed migratory blebbing, migration across porous membranes, directed migration, as well as wound healing in response to GM-CSF. Third, cell migration towards physical stimuli (specifically magnetic fields) was engineered by a system composed of CaRQ and a magnetic-sensitive gene circuit (henceforth the TF circuit). The TF circuit is composed of a chimeric TRPV1 receptor and ferritin. It was observed that a magnetic field induced migration when the TF circuit was paired with CaRQ. Taken together, this thesis demonstrates the utility of CaRQ in engineering cell migration to different stimuli in vitro, thus taking a step forward in engineering a versatile system to localize cells for cell-based therapy. It remains to be seen if these genetically-encoded systems will have efficacy in vivo.Ph.D

Possible Mechanisms for the Effects of Sound Vibration on Human Health

This paper presents a narrative review of research literature to “map the landscape” of the mechanisms of the effect of sound vibration on humans including the physiological, neurological, and biochemical. It begins by narrowing music to sound and sound to vibration. The focus is on low frequency sound (up to 250 Hz) including infrasound (1–16 Hz). Types of application are described and include whole body vibration, vibroacoustics, and focal applications of vibration. Literature on mechanisms of response to vibration is categorized into hemodynamic, neurological, and musculoskeletal. Basic mechanisms of hemodynamic effects including stimulation of endothelial cells and vibropercussion; of neurological effects including protein kinases activation, nerve stimulation with a specific look at vibratory analgesia, and oscillatory coherence; of musculoskeletal effects including muscle stretch reflex, bone cell progenitor fate, vibration effects on bone ossification and resorption, and anabolic effects on spine and intervertebral discs. In every category research on clinical applications are described. The conclusion points to the complexity of the field of vibrational medicine and calls for specific comparative research on type of vibration delivery, amount of body or surface being stimulated, effect of specific frequencies and intensities to specific mechanisms, and to greater interdisciplinary cooperation and focus

The Effects of Long-Term 40-Hz Physioacoustic Vibrations on Motor Impairments in Parkinson’s Disease: A Double-Blinded Randomized Control Trial

Recent studies have suggested that vibration therapy may have a positive influence in treating motor symptoms of Parkinson’s disease (PD). However, quantitative evidence of the benefits of vibration utilized inconsistent methods of vibration delivery, and to date there have been no studies showing a long-term benefit of 40 Hz vibration in the PD population. The objective of this study was to demonstrate the efficacy of vibration administered via a physioacoustic therapy method (PAT) on motor symptoms of PD over a longer term, completed as a randomized placebo-controlled trial. Overall motor symptom severity measured by the Unified Parkinson’s Disease Rating Scale III showed significant improvements in the treatment group over 12 weeks. Specifically, all aspects of PD, including tremor, rigidity, bradykinesia, and posture and gait measures improved. To our knowledge, this is the first study to quantitatively assess 40-Hz vibration applied using the PAT method for potential long-term therapeutic effects on motor symptoms of PD

Genetically Encoded Circuit for Remote Regulation of Cell Migration by Magnetic Fields

Magnetoreception can be generally defined as the ability to transduce the effects of a magnetic field into a cellular response. Magnetic stimulation at the cellular level is particularly attractive due to its ability for deep penetration and minimal invasiveness, allowing remote regulation of engineered biological processes. Previously, a magnetic-responsive genetic circuit was engineered using the transient receptor potential vanilloid 1 (TRPV1) and the iron containing ferritin protein (<i>i.e.</i>, the TF circuit). In this study, we combined the TF circuit with a Ca²⁺ activated RhoA protein (CaRQ) to allow a magnetic field to remotely regulate cell migration. Cells expressing the TF circuit and CaRQ exhibited consistent dynamic protrusions, leading to migration along a porous membrane, directed spreading in response to a magnetic field gradient, as well as wound healing. This work offers a compelling interface for programmable electrical devices to control the migration of living systems for potential applications in cell-based therapy

Impact of COVID-19 pandemic on rare diseases - A case study on thalassaemia patients in Bangladesh

Objectives: Thalassaemia is a life-threatening rare disease, which requires regular blood transfusion and medical care. The information on how thalassaemia patients are affected during the unprecedented COVID-19 crisis is scarce. This study aimed to assess the impact of the COVID-19 pandemic on the blood transfusion and healthcare access of thalassaemia patients at the community level in Bangladesh. Methods: A cross-sectional study was conducted among thalassaemia patients registered in a community-based ‘thalassaemia registry’ in Jamapur, Bangladesh. Results: As compared to pre-COVID-19 time, the number of blood transfusions among patients under the thalassaemia registry was significantly reduced during COVID-19 pandemic (190 units versus 81 units). In addition, the median number of red cell transfusions per patient was dropped significantly from 4 units to one unit. Over 80% of patient had no access to healthcare services at all during the early phase of the pandemic. Conclusions: Emergency response with appropriate mitigative measures must be a priority for addressing an acute shortage of blood supply in situations like COVID-19 pandemic

Demographic characteristics, clinical symptoms, biochemical markers and probability of occurrence of severe dengue : A multicenter hospital-based study in Bangladesh

Establishing reliable early warning models for severe dengue cases is a high priority to facilitate triage in dengue-endemic areas and optimal use of limited resources. However, few studies have identified the complex interactive relationship between potential risk factors and severe dengue. This research aimed to assess the potential risk factors and detect their high-order combinative effects on severe dengue. A structured questionnaire was used to collect detailed dengue outbreak data from eight representative hospitals in Dhaka, Bangladesh, in 2019. Logistic regression and machine learning models were used to examine the complex effects of demographic characteristics, clinical symptoms, and biochemical markers on severe dengue. A total of 1,090 dengue cases (158 severe and 932 non-severe) were included in this study. Dyspnoea (Odds Ratio [OR] = 2.87, 95% Confidence Interval [CI]: 1.72 to 4.77), plasma leakage (OR = 3.61, 95% CI: 2.12 to 6.15), and hemorrhage (OR = 2.33, 95% CI: 1.46 to 3.73) were positively and significantly associated with the occurrence of severe dengue. Classification and regression tree models showed that the probability of occurrence of severe dengue cases ranged from 7% (age >12.5 years without plasma leakage) to 92.9% (age ≤12.5 years with dyspnoea and plasma leakage). The random forest model indicated that age was the most important factor in predicting severe dengue, followed by education, plasma leakage, platelet, and dyspnoea. The research provides new evidence to identify key risk factors contributing to severe dengue cases, which could be beneficial to clinical doctors to identify and predict the severity of dengue early.</p

Photoactivatable intein has spatial precision.

<p>(A) HeLa cells co-expressing RhoA_N-InN-mRFP and LOVInC-RhoA_C-Venus and viewed under low magnification (20x objective). Two groups of cells that have been identified by white arrows and shown enlarged in panels B and C. Cells located near the center of the field were photostimulated with periodic interval of blue-light (1 second every 0.5 min). (D) Low magnification of the same set of cells after 150 mins of showing cells within the illumination zone (dotted white circle) have undergone dynamic blebbing while cells outside the illumination zone generally remained unchanged. Again, the two groups of cells are shown enlarged in panels E and F. Scale bars are 50 μm for A and D and 30 μm for B, C, E, and F. Images are in false colour.</p

An Engineered Split Intein for Photoactivated Protein <i>Trans</i>-Splicing

<div><p>Protein splicing is mediated by inteins that auto-catalytically join two separated protein fragments with a peptide bond. Here we engineered a genetically encoded synthetic photoactivatable intein (named LOVInC), by using the light-sensitive LOV2 domain from <i>Avena sativa</i> as a switch to modulate the splicing activity of the split DnaE intein from <i>Nostoc punctiforme</i>. Periodic blue light illumination of LOVInC induced protein splicing activity in mammalian cells. To demonstrate the broad applicability of LOVInC, synthetic protein systems were engineered for the light-induced reassembly of several target proteins such as fluorescent protein markers, a dominant positive mutant of RhoA, caspase-7, and the genetically encoded Ca²⁺ indicator GCaMP2. Spatial precision of LOVInC was demonstrated by targeting activity to specific mammalian cells. Thus, LOVInC can serve as a general platform for engineering light-based control for modulating the activity of many different proteins.</p></div

Determining the minimal functional <i>Npu</i>DnaE_C intein with PTS activity in mammalian cells.

<p>(A) Protein sequence of systematic truncations of the first N-terminus β-strand of <i>Npu</i>DnaE_C intein. (B) Schematic diagram of constructs used for establishing PTS activity, where Lyn is a plasma membrane (PM) localization sequence (shown in purple). Successful PTS activity would result in Cerulean and Venus localized to the PM while mRFP would be localized to the cytoplasm. HeLa cells expressing M-Cerulean-InN-mRFP resulted in (C) CFP and (D) RFP fluorescence localized to the PM. (D) Expression of InC-Venus resulted in YFP fluorescence distributed in the cytoplasm. Co-expression of the two constructs triggered PTS activity resulting PM localization of (F) CFP and (H) YFP while the cytoplasm is (G) RFP fluorescence. Co-expression of M-Cerulean-InN-mRFP with (I-K) t4 and (L-N) t9 truncations of N-terminus of InC in HeLa cells shown in CFP, RFP and YFP fluorescence, respectively. (O) The percentage of co-transfected cells with truncated mutants of InC (<i>n</i> = 6 experiments with over 20 cells). β-strands are shown in orange arrow. Comparison between wild-type: *<i>p <</i> 0.005, ** <i>p <</i> 0.001. Error bars are standard deviation. Scale bars are 10 μm. Images are in false colour.</p