Novel mass spectrometry-based approaches for the characterisation of systems of biological interest

Originally established as an analytical technique in the fields of physics and chemistry, mass spectrometry has now also become an essential tool in biology. Advances in ionisation methods and novel types of instrumentation have led to the development of mass spectrometry for the analysis of a wide variety of biological samples. The work presented here describes the use mass spectrometry for the study of a number of biological systems. A new family of techniques has been developed allowing ions to be created under ambient conditions. Three of these ambient ionisation techniques, coupled to different mass analysers, were employed for the rapid screening of pharmaceutical formulations. Active ingredients were identified and subjected to collisionally induced dissociation, enabling the elucidation of potential fragmentation pathways. Drug metabolites were also successfully identified from biological samples. Inorganic mass spectrometry was employed to probe the metal centres of the enzyme, particulate methane monooxygenase, a methane-oxidising complex found in certain bacteria. This protein has been extensively studied, but questions remain regarding its catalytic mechanism, particularly the involvement of indigenous metal ions. Inductively-coupled plasma mass spectrometry experiments have indicated the presence of copper and iron within the enzyme. Protein cross-sections, obtained using ion mobility mass spectrometry, can be used to probe the conformation of molecules in the gas phase. A commercial instrument was used to investigate human hemoglobin from clinical samples. A complex assembly mechanism was deduced, resolving previous disputes in the literature, and conformational differences were observed between healthy and sickle molecules. The field of proteomics is rapidly evolving; as described, techniques are constantly being developed and improved to deal with the enormous complexity that proteomes present. Three proteomics approaches were used to study a recently identified bacterium under two growth conditions. Differences in protein expression were observed and correlated to relevant biological pathways

Rainforest, Reef, and our Appetite for Beef: Communication for Sustainable Behaviour Change

Meat production, and the current rate of consumption, is one of the leading causes of tropical deforestation, freshwater degradation, ocean dead zones, and wild species extinction. The livestock sector is also a major driver of global climate change, such that meeting the 2°C climate objective will be impossible to achieve without a global shift towards a plant-based diet. Despite its gravity, the environmental impact of the meat industry is severely underrepresented in policy and communication, which has resulted in a widespread public awareness gap. The present research sought to assess whether closing this awareness gap had the potential to induce attitudinal and behavioural dietary change. In a controlled experiment, an online survey presenting an environmentally-framed essay about animal agriculture was administered to North American participants. Participants answered a series of questions assessing their response to the information. The results indicate that the environment frame, as applied to animal agriculture, was both new and valuable to most participants, with many expressing increased concern. Over half of the participants also experienced a change in dietary choice after reading the essay. These findings suggest that closing the public awareness gap, by using the environment frame to disseminate information, is an effective means of promoting dietary change

Novel mass spectrometry-based approaches for the characterisation of systems of biological interest

Originally established as an analytical technique in the fields of physics and chemistry, mass spectrometry has now also become an essential tool in biology. Advances in ionisation methods and novel types of instrumentation have led to the development of mass spectrometry for the analysis of a wide variety of biological samples. The work presented here describes the use mass spectrometry for the study of a number of biological systems. A new family of techniques has been developed allowing ions to be created under ambient conditions. Three of these ambient ionisation techniques, coupled to different mass analysers, were employed for the rapid screening of pharmaceutical formulations. Active ingredients were identified and subjected to collisionally induced dissociation, enabling the elucidation of potential fragmentation pathways. Drug metabolites were also successfully identified from biological samples. Inorganic mass spectrometry was employed to probe the metal centres of the enzyme, particulate methane monooxygenase, a methane-oxidising complex found in certain bacteria. This protein has been extensively studied, but questions remain regarding its catalytic mechanism, particularly the involvement of indigenous metal ions. Inductively-coupled plasma mass spectrometry experiments have indicated the presence of copper and iron within the enzyme. Protein cross-sections, obtained using ion mobility mass spectrometry, can be used to probe the conformation of molecules in the gas phase. A commercial instrument was used to investigate human hemoglobin from clinical samples. A complex assembly mechanism was deduced, resolving previous disputes in the literature, and conformational differences were observed between healthy and sickle molecules. The field of proteomics is rapidly evolving; as described, techniques are constantly being developed and improved to deal with the enormous complexity that proteomes present. Three proteomics approaches were used to study a recently identified bacterium under two growth conditions. Differences in protein expression were observed and correlated to relevant biological pathways.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Study of role of blood transfusion in obstetric emergencies

Background: Blood Transfusion is recognized as one of the eight essential component of comprehensive emergency obstetric care which has been shown to reduce the maternal mortality.1,2 In developing country like India, efforts should be done to make blood and transfusion services well maintained and quickly available to reduce maternal morbidity from haemorrhage and thus decrease the incidence of maternal mortality. Aims & objectives: (1) To study clinical status of the patients at time of admission. (2) To study the role of antenatal visits in all patients required blood transfusion. (3) To study the effect of blood components on the patients’ health. (4) To screen out the patients of high risk pregnancy and treat them safely. (5) To study causes of maternal mortality.Methods: Retrospective study of requirement of blood transfusion in antenatal and postnatal patients who came in labour room during last 3 month period at tertiary care Centre, Ahmedabad.Results: during the whole study out of 2200 patients 440 patients required blood transfusion among which 70% required due to obstetric hemorrhage and 30% due to severe anemia (less than 7 gm/dl). Major associated complications in the transfused patients were anemia (34%) and PPH (36%). 4 patients expired among them 2 were due to development of DIC and septicemia, 1 due to severe anemia and 1 due to severe PPH.Conclusions: Ensuring a safe supply of blood and blood products and the appropriate and rational clinical use of blood. Strategies made to maximize the haemoglobin (Hb) level at the time of delivery as well as to minimize blood loss. Active management of the third stage of labour is required to prevent avoidable morbidities, such as PPH, Retained product of conception, and vaginal lacerations.

Comparative study of bupivacaine alone and bupivacaine along with buprenorphine in axillary brachial plexus block: a prospective, randomized, single blind study

Background: Different additives have been used to prolong brachial plexus block. We performed a prospective, randomized single-blind study to compare Bupivacaine alone and Bupivacaine along with Buprenorphine for onset, quality, and duration of block as well as post-operative analgesia and any complication in axillary brachial- plexus block.Methods: Randomized controlled study was carried out among 60 patients of either sex, aged 20-60 years. ASA grade I or II undergoing elective hand, forearm, elbow surgery under axillary brachial plexus block. Patients were randomly divided into two groups.Group-l received 30 ml of 0.35% Bupivacaine alone in axillary block.Group-II received 30 ml of 0.35% Bupivacaine with 3µg/kg Buprenorphine in axillary block. Time taken for onset and completion of motor and sensory block as well as complete duration of block were noted in both groups. Any complication during procedure, during surgery as well as post-operatively were noted and treated.Results: Addition of Buprenorphine (3µg/kg) to Bupivacaine mixture in peripheral nerve block did not affected the onset time for motor as well as sensory block. Mean duration of motor block was 284.33±78.94 mins. in group I and in group II 307.33±60.26 mins. Mean duration of sensory block 305.066±83.64 mins. in group I while 580.166±111.45 mins. in group II. It suggests duration of sensory block was prolonged in group II then group I.Conclusions: Addition of Buprenorphine to local anesthetic drug provides good post-operative analgesia. Buprenorphine significantly prolongs sensory block and lengthens duration of analgesia without prolonging duration of motor block

Entrepreneurial Actors in Transport Systems. An Energy Cultures Perspective

Moving to a low carbon economy in New Zealand requires a considerable shift in the current transport system as a whole (Carrington et al, 2014). Indeed, the New Zealand transportation system in has a number of key issues including old vehicle stock, old inefficient ICE engines, lack of rail infrastructure investment, low petrol taxes relative to OECD countries and a focus on road building (Vivid Economics & University of Auckland Business School, 2012). Given the complexity involved, shifting the current transport socio-technical systems (with the reliance on the ICE) will be difficult. Starting a business that relies on shifting such an embedded regime can thus be a hard journey. This report looks at the entrepreneurs operating businesses that challenge the mainstream and dominant transportation regime in New Zealand. They can be considered brave people by introducing new products and services into the market that challenge the reliance on fossil fuels in transportation. This report explores each entrepreneurial venture to understand their operations, what alternatives is being offered and what the barriers are to that venture. With barriers comes opportunities and these entrepreneurs are seeing and seizing opportunities that may have the potential together to shift the regime from its current form into something that is more sustainable for the future

Feminist mobilisation for policy change on violence against women: insights from Asia

This article explores how women’s movements in China, India and Indonesia have mobilized to influence processes of legal reform on violence against women (VAW). Legal change is a complex and iterative process, in which both state and non-state actors negotiate and bargain over the content of law in the ‘policy space’, bringing different interests and needs to bear. The three countries featured here differ in many ways, including population size, political system (including varying levels and degrees of democratization and decentralization, and regional and local autonomy), and diversity in the population, including ethnicities and religions. A comparative study such as this offers important potential for understanding policy change on VAW, the role of women’s movements in this, and the obstacles to change

Lower locus coeruleus integrity in older COVID-19 survivors: initial findings from an international 7T MRI consortium

Background: The SARS-CoV-2 coronavirus has been associated with structural brain changes, consistent with its neurological manifestations. Recent studies showed a specific predilection for brainstem glial activation and hypometabolism, possibly indicating involvement of the locus coeruleus. The locus coeruleus (LC) modulates many cognitive functions and behaviors and its norepinephrine projections regulate both immune responses and vascular reactivity. We aimed to examine differences in LC integrity between COVID-19 survivors and controls. Method: Participants are enrolled across 3 US and 1 UK sites using harmonized cognitive and 7T MR-imaging protocols. Here, we analyzed data from 18 participants enrolled at Houston Methodist (12 COVID-19 survivors, 6 controls; Figure 1). COVID-19 survivors were required to have had a positive antigen test and an illness syndrome consistent with COVID-19. Healthy controls were required to have no significant pre-existing medical, neurologic, or psychiatric illness and no illness requiring hospitalization in the last 2 years. LC imaging was performed using a dedicated 7T MT-TFL sequence (0.4 x 0.4 x0.5mm). A site-specific normalized template was constructed using ANTs/FSL. The entire average LC integrity as well as voxel-wise integrity values were compared between COVID-19 survivors and controls using a robust linear regression (age-controlled and threshold free cluster enhancement corrected). LC integrity was correlated with age, sex, ethnicity and cognition using Spearman’s rank correlation. Result: Average LC integrity was not correlated with age, sex, or Hispanic ethnicity (p\u3e0.3). COVID-19 survivors did not differ from Controls when examining the entire LC (p=0.54). Voxel-wise analyses revealed a small cluster (19 voxels) in the middle portion of the left LC where COVID-19 survivors exhibited lower LC integrity than controls (p=0.005; Figure 2). Integrity of this cluster was not related to age or Hispanic ethnicity (p=0.9). LC integrity did not correlate with cognitive performance within the COVID-19 survivors (Trail Making Test B: p=0.43; Craft Story delayed recall p=0.47; MoCA p=0.84). Conclusion: Consistent with previous animal and human studies, our initial findings provide evidence for neuroinvasive potential of SARS-CoV-2 localized in the middle LC. In the future, we aim to expand our sample and link these observations to the neurocognitive sequelae of COVID-19

Investigating white matter hyperintensities in a multicenter COVID-19 study using 7T MRI

Background: Emerging evidence indicates that COVID-19 can negatively impact patient’s brain health (Douaud et al., 2022) (Cecchetti et al., 2022). Common clinical symptoms include brain fog, headaches, difficulty concentrating, and loss of sense of smell or taste. Some studies suggest that SARS-CoV-2 infection can damage the blood brain barrier either directly or through immune-inflammatory mechanisms (Zhang, et al. 2021). White matter hyperintensities (WMH) are imaging biomarkers of brain vascular or inflammatory injury. We investigated the association between severity of COVID-19 infection and burden of white matter hyperintensity volumes within a diverse multi-nation, multi-racial cohort using 7 Tesla (7T) MRI that can detect more subtle injury than conventional 1.5 or 3T MRI. Method: Participants were recruited at 4 sites: Pittsburgh, San Antonio and Houston, USA, and Nottingham, UK. To date, we have scanned and included the following participants in our analysis (Table 1). Detailed cognitive, neurological, mood and functional assessments and high-resolution MRI scans were collected. Subsequent WMH segmentation was performed using our in-house built deep learning based model (Figure 1). All segmentations were visually inspected and manually corrected before statistical analysis. Normalized WMH is calculated as a ratio of the WMH volume and the intracranial volume (WMH/ICV). Imaging data for an additional 36 age-matched controls were retrieved from the 7 Tesla Bioengineering Research Program (7TBRP) imaging bank at Pittsburgh. Result: Figure 1 shows the WMH segmentation outputs from our deep learning based model on images acquired at the 3 sites. Our Linear regression models along with our non-parametric Kruskal-Wallis test result suggests that compared to mild COVID cases and healthy control, COVID infected individuals that were ICU admitted show elevated WMH burden (Figure 2). Conclusion: Our results demonstrate that white matter hyperintensity volumes were higher among patients who had severe acute COVID infection that required ICU admission, compared to healthy age-matched controls. In contrast, no difference in white matter burden was observed in patients with mild COVID infection compared to healthy controls. Additional data (both cross-sectional and longitudinal), including more sensitive MRI measures is being collected to define the full spectrum of brain injury associated with sequelae of COVID infection

Identificaction and quantificationof proteins from Methylophaga Thiooxidans and Methylocella Silvestris using label-free LC/MS

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