Antibody-mediated initiation and lymphocyte-targeting therapies in CNS demyelinating disease

Multiple sclerosis is inflammatory central nervous system disease, characterised by areas of demyelination and axonal loss. The pathogenic mechanism behind the disease still remains unknown, however it is thought to be mainly T cell-mediated. Notwithstanding this, B cells have increasingly been recognized as key mediators of disease. This work focuses on three distinct characteristics of MS pathology in the animal model experimental autoimmune encephalomyelitis (EAE). The first project focused on establishing inhibition of Bruton´s tyrosine kinase (BTK) as a novel therapeutic approach. BTK is centrally placed in B cell receptor (BCR) signalling. In a B cell-mediated EAE model, induced by injection of MOG protein, we observed that evobrutinib, a novel BTK inhibitor, dose-dependently reduced clinical disease. Evobrutinib inhibited BCR-mediated phenotypic maturation of B cells from follicular (FO) II to FO I and reduced activation of B cells and T cells. It diminished calcium mobilization and cytokine production after BCR stimulation in murine and human B cells. Investigating MS patients, we did not observe a difference in B cell frequency, BTK expression or phosphorylation of BTK after BCR stimulation. Taken together, we demonstrated that BTK inhibition (BTKi) is a promising new strategy to control pathogenic B cell activity in a model of CNS autoimmunity. The second project investigated the effects of long-term high dose vitamin D supplementation on the peripheral immune system and EAE severity. We observed clinical and histological deterioration of EAE after long-term high dose supplementation of vitamin D. Further investigations traced this effect to a secondary hypercalcemia, which in contrast to vitamin D, increased the activation and differentiation of T cells both in vitro and in vivo. Since MS patients are often continuously supplemented with vitamin D over long periods of time, our work cautions patients and clinicians to be attentive of potential side effects by hypercalcemia. The third project focused on antibodies in the initiation of disease. We investigated the capacity of myelin-reactive antibodies to facilitate encephalitogenic responses via opsonisation of CNS antigen. We observed that antibody production in the absence of B cells was sufficient to induce EAE in a transgenic mouse model. Additionally, adoptive transfer of antibodies in mice containing MOG-specific T cells induced disease via otherwise unresponsive myeloid antigen-presenting cells (APCs). MOG-targeting antibodies enabled Fcreceptor (FcR) mediated recognition and phagocytosis in in vitro differentiated macrophages. Additionally, antibody preparations from neuromyelitis optica patients positive for MOG antibodies similarly facilitated recognition by myeloid APCs. These results establish opsonisation of CNS antigen by specific antibodies as a novel mechanism to trigger CNS demyelination.2020-07-0

A PLL-Based Frequency Shift Measurement System for Chemical and Biological Sensing

A PLL-based frequency shift measurement system for chemical and biological sensing was developed and implemented in the form of two discrete electronic assemblies. One of the assemblies consists of a VCO which contains a microwave resonator sensor while the other assembly contains commercially available PLL and MCU devices, as well as various other discrete components. When mated together, a PLL-based frequency synthesizer is realized, the output frequency of which is ~4.5 GHz. The system is used to measure the frequency shift exhibited by the frequency synthesizer when several commonly-known chemical substances are applied to the microwave resonator sensor test fixture. Because the amount of measured frequency shift is proportional to the dielectric constant of a given material under test (MUT), this system can potentially be used as part of a chemical identification system. This measurement system is also attractive in that it represents a stand-alone or 'self-contained' system which does not require usage of any additional expensive and bulky electronic diagnostic equipment such as a network analyzer or signal generator, making it a relatively inexpensive and portable solution. Attempts to use the system to measure frequency shift resulting from application of various common chemical substances to the sensor fixture results in derivation of dielectric constant values which hold very close agreement (+/-2%) to the published/theoretical dielectric constant values for each respective chemical substance

The Future of First Amendment Overbreadth

In Broadrick, Justice White suggests that the willingness of the Court in the past to accord standing to litigants to raise the over-broad aspects of a statute without regard to their own conduct\u27depended on a judicial prediction or assumption regarding the threat to liberty that the statute posed.\u27 The spirit with which the Court makes such predictions would appear to be determinative not only of the underlying standing issue but of the overbreadth claimas well. The key to discovering the paths by which the Court is retreating from its overbreadth holiday of the sixties lies in the recognition that the central dynamic of overbreadth is the peril posed by standardless administration rather than the threat of a chilling effect on first amendment rights. That recognition enables one to see that past cases are cluttered with sources other than the statutes themselves that were searched by the Court in an effort to predict whether the danger of overbreadth was great enough to call for radical treatment

"What do you Mean I Cannot Consent For My Grandmother's Medical Procedure?": Key Issues With State Default Surrogate Decision Making Laws

When a patient is unable to make medical decisions, a health care surrogate must be designated to make decisions on the patient's behalf. Studies show that fewer than 20% of patients have completed health care representative forms to legally designate a surrogate. Without a prior designation, surrogates are determined via state statute. Currently, there is no up-to-date comprehensive evaluation of state surrogate legislation. A survey of state legislative codes was conducted to determine: 1) whether the state has a default surrogate statute; 2) who is included as an acceptable legal surrogate; and 3) whether there is a hierarchy to determine a final decision-maker. Currently, 36 states have enacted some form of surrogate statute. There is little consistency between states regarding who may serve as a surrogate decision- maker. The key challenges with state laws include: 1) a narrow list of persons who qualify as allowable legal surrogates; and 2) a lack of a hierarchy to determine a final decision-maker. The results of this survey show that state surrogate decision making laws have many flaws which could affect patient care. The narrow construction of state laws can leave patients in situations where they either have no qualified surrogate under the law, or where they have multiple surrogates with competing interests who may be unable to reach consensus on the patient's medical care. State laws need to be changed so that they accurately reflect the realities of clinical practice and expanded to allow a broader spectrum of potential surrogates

Older Adults and Forgoing Cancer Screening

Although there is a growing recognition that older adults and those with extensive comorbid conditions undergo cancer screening too frequently, there is little information about patients’ perceptions regarding cessation of cancer screening. Information on older adults’ views of screening cessation would be helpful both for clinicians and for those designing interventions to reduce overscreening