Exploring Structure-Dynamics-Function Relationship in Proteins, Protein: Ligand and Protein: Protein Systems through Computational Methods

The study focuses on understanding the dynamic nature of interactions between molecules and macromolecules. Molecular modeling and simulation technologies are employed to understand how the chemical constitution of the protein, specific interactions and dynamics of its structure provide the basis of its mechanism of function. The structure-dynamics-function relationship is investigated from quantum to macromolecular-assembly level, with applications in the field of rationale drug discovery and in improving efficiency of renewable sources of energy. Results presented include investigating the role of dynamics in the following: 1) In interactions between molecules: analyzing dynamic nature of a specific non-covalent interaction known as “anion-π [pi]” in RmlC protein. 2) In interactions between molecules and macromolecules: defining the structural basis of testosterone activation of GPRC6A. 3) In disrupting the function using specific substrate interactions: incorporating protein dynamics and flexibility in structure-based drug-discovery approach targeting the prothrombinase coagulation complex. 4) In interactions between macromolecules: elucidating the protein-protein binding and dynamics of electron-transport proteins, Ferrodoxin and Cytochrome c6, with Cyanobacterial Photosystem I

Micro-motion controller

Micro-motions in surgical applications are small motions in the range of a few millimeters and are common in ophthalmic surgery, neurosurgery, and other surgeries which require precise manipulation over short distances. Robotic surgery is replacing traditional open surgery at a rapid pace due to the obvious health benefits, however, most of the robotic surgical tools use robotic motion controllers that are designed to work over a large portion of the human body, thus involving motion of the entire human arm at shoulder joint. This requirement to move a large inertial mass results in undesirable, unwanted, and imprecise motion. This senior design project has created a 2-axis micro-motion “capable” platform, where the device studies the most common linear, 2-D surgical micro-motion of pinched human fingers in a damped and un-damped state. Through a system of printed and modeled parts in combination with motors and encoders a microsurgical controller was developed which can provide location-based output on a screen. Mechanical damping was introduced to research potential stability of micro-motion in any surgeon’s otherwise unsteady hand. The device is to also serve as a starter set for future biomedical device research projects in Santa Clara University’s bioengineering department. Further developments in the microsurgical controller such as further scaling, addition of a third axis, haptic feedback through the microcontroller, and component encasing to allow productization for use on an industrial robotic surgical device for clinical applications

Discovery of Novel Nonactive Site Inhibitors of the Prothrombinase Enzyme Complex

© 2016 American Chemical Society. The risk of serious bleeding is a major liability of anticoagulant drugs that are active-site competitive inhibitors targeting the Factor Xa (FXa) prothrombin (PT) binding site. The present work identifies several new classes of small molecule anticoagulants that can act as nonactive site inhibitors of the prothrombinase (PTase) complex composed of FXa and Factor Va (FVa). These new classes of anticoagulants were identified, using a novel agnostic computational approach to identify previously unrecognized binding pockets at the FXa-FVa interface. From about three million docking calculations of 281 128 compounds in a conformational ensemble of FXa heavy chains identified by molecular dynamics (MD) simulations, 97 compounds and their structural analogues were selected for experimental validation, through a series of inhibition assays. The compound selection was based on their predicted binding affinities to FXa and their ability to successfully bind to multiple protein conformations while showing selectivity for particular binding sites at the FXa/FVa interface. From these, thirty-one (31) compounds were experimentally identified as nonactive site inhibitors. Concentration-based assays further identified 10 compounds represented by four small-molecule families of inhibitors that achieve dose-independent partial inhibition of PTase activity in a nonactive site-dependent and self-limiting mechanism. Several compounds were identified for their ability to bind to protein conformations only seen during MD, highlighting the importance of accounting for protein flexibility in structure-based drug discovery approaches

Ensemble-based docking: From hit discovery to metabolism and toxicity predictions

This paper describes and illustrates the use of ensemble-based docking, i.e., using a collection of protein structures in docking calculations for hit discovery, the exploration of biochemical pathways and toxicity prediction of drug candidates. We describe the computational engineering work necessary to enable large ensemble docking campaigns on supercomputers. We show examples where ensemble-based docking has significantly increased the number and the diversity of validated drug candidates. Finally, we illustrate how ensemble-based docking can be extended beyond hit discovery and toward providing a structural basis for the prediction of metabolism and off-target binding relevant to pre-clinical and clinical trials

Combining Ascochyta blight and Botrytis grey mould resistance in chickpea through interspecific hybridization

Ascochyta blight (AB) caused by Ascochyta rabiei (Pass.) Labr. and Botrytis grey mould (BGM) caused by Botrytis cinerea (Pers. ex Fr.) are important diseases of the aerial plant parts of chickpea in most chickpea growing areas of the world. Although conventional approaches have contributed to reducing disease, the use of new technologies is expected to further reduce losses through these biotic stresses. Reliable screening techniques were developed: ‘field screening technique’ for adult plant screening, ‘cloth chamber technique’ and ‘growth chamber technique’ for the study of races of the pathogen and for segregating generations. Furthermore, the ‘cut twig technique’ for interspecific population for AB and BGM resistance was developed. For introgression of high levels of AB and BGM resistance in cultivated chickpea from wild relatives, accessions of seven annual wild Cicer spp. were evaluated and identified: C. judaicum accessions 185, ILWC 95 and ILWC 61, C. pinnatifidum accessions 188, 199 and ILWC 212 as potential donors. C. pinnatifidum accession188 was crossed with ICCV 96030 and 62 F9 lines resistant to AB and BGM were derived. Of the derived lines, several are being evaluated for agronomic traits and yield parameters while four lines, GL 29029, GL29206, GL29212, GL29081 possessing high degree of resistance were crossed with susceptible high yielding cultivars BG 256 to improve resistance and to undertake molecular studies. Genotyping of F2 populations with SSR markers from the chickpea genome was done to identify markers potentially linked with AB and BGM resistance genes. In preliminary studies, of 120 SSR markers used, six (Ta 2, Ta 110, Ta 139, CaSTMS 7, CaSTMS 24 and Tr 29) were identified with polymorphic bands between resistant derivative lines and the susceptible parent. The study shows that wild species of Cicer are the valuable gene pools of resistance to AB and BGM. The resistant derivative lines generated here can serve as good pre-breeding material and markers identified can assist in marker assisted selection for resistance breeding

Combining Ascochyta blight and Botrytis grey mould resistance in chickpea through interspecific hybridization

Ascochyta blight (AB) caused by Ascochyta rabiei (Pass.) Labr. and Botrytis grey mould (BGM) caused by Botrytis cinerea (Pers. ex Fr.) are important diseases of the aerial plant parts of chickpea in most chickpea growing areas of the world. Although conventional approaches have contributed to reducing disease, the use of new technologies is expected to further reduce losses through these biotic stresses. Reliable screening techniques were developed: ‘field screening technique’ for adult plant screening, ‘cloth chamber technique’ and ‘growth chamber technique’ for the study of races of the pathogen and for segregating generations. Furthermore, the ‘cut twig technique’ for interspecific population for AB and BGM resistance was developed. For introgression of high levels of AB and BGM resistance in cultivated chickpea from wild relatives, accessions of seven annual wild Cicer spp. were evaluated and identified: C. judaicum accessions 185, ILWC 95 and ILWC 61, C. pinnatifidum accessions 188, 199 and ILWC 212 as potential donors. C. pinnatifidum accession188 was crossed with ICCV 96030 and 62 F9 lines resistant to AB and BGM were derived. Of the derived lines, several are being evaluated for agronomic traits and yield parameters while four lines, GL 29029, GL29206, GL29212, GL29081 possessing high degree of resistance were crossed with susceptible high yielding cultivars BG 256 to improve resistance and to undertake molecular studies. Genotyping of F2 populations with SSR markers from the chickpea genome was done to identify markers potentially linked with AB and BGM resistance genes. In preliminary studies, of 120 SSR markers used, six (Ta 2, Ta 110, Ta 139, CaSTMS 7, CaSTMS 24 and Tr 29) were identified with polymorphic bands between resistant derivative lines and the susceptible parent. The study shows that wild species of Cicer are the valuable gene pools of resistance to AB and BGM. The resistant derivative lines generated here can serve as good pre-breeding material and markers identified can assist in marker assisted selection for resistance breeding

Neurovisceral phenotypes in the expression of psychiatric symptoms

This review explores the proposal that vulnerability to psychological symptoms, particularly anxiety, originates in constitutional differences in the control of bodily state, exemplified by a set of conditions that include Joint Hypermobility, Postural Tachycardia Syndrome and Vasovagal Syncope. Research is revealing how brainbody mechanisms underlie individual differences in psychophysiological reactivity that can be important for predicting, stratifying and treating individuals with anxiety disorders and related conditions. One common constitutional difference is Joint Hypermobility, in which there is an increased range of joint movement as a result of a variant of collagen. Joint hypermobility is over-represented in people with anxiety, mood and neurodevelopmental disorders. It is also linked to stress-sensitive medical conditions such as irritable bowel syndrome, chronic fatigue syndrome and fibromyalgia. Structural differences in 'emotional' brain regions are reported in hypermobile individuals, and many people with joint hypermobility manifest autonomic abnormalities, typically Postural Tachycardia Syndrome. Enhanced heart rate reactivity during postural change and as recently recognised factors causing vasodilatation (as noted post prandially, post exertion and with heat) is characteristic of Postural Tachycardia Syndrome, and there is a phenomenological overlap with anxiety disorders, which may be partially accounted for by exaggerated neural reactivity within ventromedial prefrontal cortex. People who experience Vasovagal Syncope, a heritable tendency to fainting induced by emotional challenges (and needle/blood phobia), are also more vulnerable to anxiety disorders. Neuroimaging implicates brainstem differences in vulnerability to faints, yet the structural integrity of the caudate nucleus appears important for the control of fainting frequency in relation to parasympathetic tone and anxiety. Together there is clinical and neuroanatomical evidence to show that common constitutional differences affecting autonomic responsivity are linked to psychiatric symptoms, notably anxiety

Impact of opioid-free analgesia on pain severity and patient satisfaction after discharge from surgery: multispecialty, prospective cohort study in 25 countries

Background: Balancing opioid stewardship and the need for adequate analgesia following discharge after surgery is challenging. This study aimed to compare the outcomes for patients discharged with opioid versus opioid-free analgesia after common surgical procedures.Methods: This international, multicentre, prospective cohort study collected data from patients undergoing common acute and elective general surgical, urological, gynaecological, and orthopaedic procedures. The primary outcomes were patient-reported time in severe pain measured on a numerical analogue scale from 0 to 100% and patient-reported satisfaction with pain relief during the first week following discharge. Data were collected by in-hospital chart review and patient telephone interview 1 week after discharge.Results: The study recruited 4273 patients from 144 centres in 25 countries; 1311 patients (30.7%) were prescribed opioid analgesia at discharge. Patients reported being in severe pain for 10 (i.q.r. 1-30)% of the first week after discharge and rated satisfaction with analgesia as 90 (i.q.r. 80-100) of 100. After adjustment for confounders, opioid analgesia on discharge was independently associated with increased pain severity (risk ratio 1.52, 95% c.i. 1.31 to 1.76; P < 0.001) and re-presentation to healthcare providers owing to side-effects of medication (OR 2.38, 95% c.i. 1.36 to 4.17; P = 0.004), but not with satisfaction with analgesia (beta coefficient 0.92, 95% c.i. -1.52 to 3.36; P = 0.468) compared with opioid-free analgesia. Although opioid prescribing varied greatly between high-income and low- and middle-income countries, patient-reported outcomes did not.Conclusion: Opioid analgesia prescription on surgical discharge is associated with a higher risk of re-presentation owing to side-effects of medication and increased patient-reported pain, but not with changes in patient-reported satisfaction. Opioid-free discharge analgesia should be adopted routinely