Exploring the role of miR-34a in regulating adipose inflammation during obesity

Background: Obesity is not a new disease, with roots that can be traced back to 400 BC. However, with the staggering increase in individuals that are overweight and obese since the 1980s, now over a quarter of individuals in Europe and the Americas are classed as obese. This presents a global health problem that needs to be addressed with novel therapies. It is now well accepted that obesity is a chronic, low-grade inflammatory condition that could predispose individuals to a number of comorbidities. Obesity is associated with cardiovascular diseases (CVDs) and type 2 diabetes (T2D) as part of “the metabolic syndrome,” and as first identified by Dr Vauge, central distribution of white adipose tissue (WAT) is an important risk factor in the development of these diseases. Subsequently, visceral WAT (vWAT) was shown to be an important factor in this association with CVDs and T2D, and increasing inflammation. As the obese WAT expands, mainly through hypertrophy, there is an increase in inflammation that recruits numerous immune cells to the tissue that further exacerbate this inflammation, causing local and systemic inflammatory and metabolic effects. One of the main types of immune cell involved in this pathogenic process is pro-inflammatory M1 adipose tissue macrophages (ATMs). MicroRNAs (miRNAs) are a species of small RNAs that post-transcriptionally regulate gene expression by targeting gene mRNA, causing its degradation or translational repression. These miRNAs are promiscuous, regulating numerous genes and pathways involved in a disease, making them useful therapeutic targets, but also difficult to study. miR-34a has been shown to increase in the serum, liver, pancreas, and subcutaneous (sc)WAT of patients with obesity, non- alcoholic fatty liver disease (NAFLD) and T2D. Additionally, miR-34a has been shown to regulate a number of metabolic and inflammatory genes in numerous cell types, including those in macrophages. However, the role of miR-34a in regulating vWAT metabolism and inflammation is poorly understood. Hypothesis: miR-34a is dysregulated in the adipose tissue during obesity, causing dysregulation of metabolic and inflammatory pathways in adipocytes and ATMs that contribute to adipose inflammation and obesity’s comorbidities, particularly T2D. Method/Results: The role of miR-34a in adipose inflammation was investigated using a murine miR-34a-/- diet-induced obesity model, and primary in vitro models of adipocyte differentiation and inflammatory bone marrow-derived macrophages (BMDMs). miR-34a was shown to be ubiquitously expressed throughout the murine epididymal (e)WAT of obese high-fat diet (HFD)-fed WT mice and ob/ob mice, as well as omental WAT from patients with obesity. Additionally, miR-34a transcripts were increased in the liver and brown adipose tissue (BAT) of ob/ob and HFD-fed WT mice, compared to WT controls. When miR-34a-/- mice were fed HFD ad libitum for 24 weeks they were significantly heavier than their WT counterparts by the end of the study. Ex vivo examinations showed that miR-34a-/- eWAT had a smaller adipocyte area on chow, which significantly increased to WT levels during HFD-feeding. Additionally, miR-34a-/- eWAT showed basal increases in cholesterol and fatty acid metabolism genes Cd36, Hmgcr, Lxrα, Pgc1α, and Fasn. miR-34a-/- iBAT showed basal reductions in Cebpα and Cebpβ, with increased Pgc1α expression during HFD- feeding. The miR-34a-/- liver additionally showed increased basal transcript expression of Pgc1α, suggesting miR-34a may broadly regulate PGC1α. Accompanying the ex vivo changes in cholesterol and fatty acid metabolism genes, in vitro miR-34a-/- white adipocytes showed increased lipid content. An F4/80high macrophage population was identified in HFD-fed miR-34a-/- eWAT, with increased Il-10 transcripts and serum IL-5 protein. Following these ex vivo observations, BMDMs from WT mice upregulated miR-34a expression in response to TNFα stimulation. Additionally, miR-34a-/- BMDMs showed an ablated CXCL1 response to TNFα. Conclusion: These findings suggest miR-34a has a multi-factorial role in controlling a susceptibility to obesity, by regulating inflammatory and metabolic pathways, potentially through regulation of PGC1α.

The dehydrogenation reactions between LiH and organic amines for solid state hydrogen storage systems

Hydrogen is considered as an appealing alternative to fossil fuels in the pursuit of sustainable, secure and prosperous growth in the UK and abroad. However there exists a persisting bottleneck in the effective storage of hydrogen for mobile applications in order to facilitate a wide implementation of hydrogen fuel cells in the fossil fuel dependent transportation industry. To address this issue, new means of solid state chemical hydrogen storage are proposed in this thesis. This involves the coupling of LiH with three different organic amines: melamine, urea and dicyandiamide. In principle, thermodynamically favourable hydrogen release from these systems proceeds via the deprotonation of the protic N-H moieties by the hydridic metal hydride. Simultaneously hydrogen kinetics is expected to be enhanced over heavier hydrides by incorporating lithium ions in the proposed binary hydrogen storage systems. Whilst the concept has been successfully demonstrated by the results obtained in this work, it was observed that optimising the ball milling conditions is central in promoting hydrogen desorption in the proposed systems. The theoretical amount of 6.97 wt% by dry mass of hydrogen was released when heating a ball milled mixture of LiH and melamine (6:1 stoichiometry) to 320 °C. It was observed that ball milling introduces a disruption in the intermolecular hydrogen bonding network that exists in pristine melamine. This effect extends to a molecular level electron redistribution observed as a function of shifting IR bands. It was postulated that stable phases form during the first stages of dehydrogenation which contain the triazine skeleton. Dehydrogenation of this system yields a solid product Li2NCN, which has been rehydrogenated back to melamine via hydrolysis under weak acidic conditions. On the other hand, the LiH and urea system (4:1 stoichiometry) desorbed approximately 5.8 wt% of hydrogen, from the theoretical capacity of 8.78 wt% (dry mass), by 270 °C accompanied by undesirable ammonia and trace amount of water release. The thermal dehydrogenation proceeds via the formation of Li(HN(CO)NH2) at 104.5 °C; which then decomposes to LiOCN and unidentified phases containing C-N moieties by 230 °C. The final products are Li2NCN and Li2O (270 °C) with LiCN and Li2CO3 also detected under certain conditions. It was observed that ball milling can effectively supress ammonia formation. Furthermore results obtained from energetic ball milling experiments have indicated that the barrier to full dehydrogenation between LiH and urea is principally kinetic. Finally the dehydrogenation reaction between LiH and dicyandiamide system (4:1 stoichiometry) occurs through two distinct pathways dependent on the ball milling conditions. When ball milled at 450 RPM for 1 h, dehydrogenation proceeds alongside dicyandiamide condensation by 400 °C whilst at a slower milling speed of 400 RPM for 6h, decomposition occurs via a rapid gas desorption (H2 and NH3) at 85 °C accompanied by sample foaming. The reactant dicyandiamide can be generated by hydrolysis using the product Li2NCN.

Socio-economic status, growth, physical activity and fitness: the Madeira growth study

Within a country social conditions change over time and these conditions vary from country to country. The associations between these conditions, somatic growth, physical activity and fitness reflect these changes. Aim: The study documented variation in somatic growth, physical activity and fitness associated with socio-economic status (SES). Subjects and methods: The study involved 507 subjects (256 boys and 251 girls) from the Madeira Growth Study, a mixed longitudinal study of five cohorts (8, 10, 12, 14 and 16 years of age) followed at yearly intervals over 3 years (1996–1998). A total of 1493 observations were made. Anthropometric measurements included lengths, body mass, skeletal breadths, girths and skinfolds. Physical activity and SES were collected via questionnaire and interview. Physical fitness was assessed using the Eurofit test battery. Variation in somatic growth, physical activity and physical fitness by SES (high, average and low) was tested with analysis of variance. Results: Significant differences between SES groups were observed for height, body mass and skinfolds. Boys and girls from high SES groups were taller, heavier and fatter (subscapular and triceps skinfolds) than their peers from average and low SES groups. At some age intervals, the high SES group had larger skeletal breadths (girls) and girths (boys and girls) than low SES. Small SES differences were observed for physical activity (sport and leisure-time indices). SES was significantly associated with physical fitness. At some age levels, boys from the low SES group performed better for muscular and aerobic endurance whereas girls from the high SES group performed better for power. Conclusion: Considerable variation in somatic growth and physical fitness in association with SES has been demonstrated, but little association was found for physical activity.

Short-term secular change in height, body mass and Tanner-Whitehouse 3 skeletal maturity of Madeira youth, Portugal

Secular trends in height and weight are reasonably well documented in Europe. Corresponding observations for skeletal maturation are lacking. Aim: To assess secular trends in height, body mass and skeletal maturity of Portuguese children and adolescents and to provide updated reference values for skeletal maturity scores (SMSs). Subjects and methods: Data for 2856 children and adolescents of 4–17 years, 1412 boys and 1444 girls, from The ‘Madeira Growth Study’ (MGS; 1996–1998) and from the‘Healthy Growth of Madeira Children Study’ (CRES; 2006) were used. Height and body mass were measured. Skeletal maturity was assessed with the Tanner-Whitehouse 2 and 3 methods. Results: Children from CRES were taller and heavier than peers from MGS. Differences in height reached 5.8cm in boys and 5.5cm in girls. RUS SMSs did not differ consistently between surveys boys, while higher RUS scores were observed in CRES girls. Adult RUS SMSs for MGS and CRES combined were attained at 15.8 years in boys and 14.8 years in girls. Corresponding ages for adult Carpal SMSs were 14.4 and 14.0, respectively. Conclusion: The short-term trends for height and mass were not entirely consistent with the trends in RUS and Carpal SMSs and SAs.

First reproductive and productive results on “Ribatejano pig”

The present trail aimed to study the effect of crossbreeding between Alentejano (AL) and Bísaro (BI) swine breeds (“Ribatejano pig”) on some reproductive and productive traits. Nine AL gilts and sows and six BI gilts were crossed with BI and AL boars, respectively. Mating and farrowing dates, prolificacy and litter size at 28d were registered for all sows. The pregnancy length of AL sows was shorter (111±0.4d vs 113.7±0.5d; p=0.002) than the observed on BI females. The BI gilts presented higher prolificacy rate than AL on both total born (11.0±1.0 vs 6.7±0.8; P=0.004) and born alive piglets (10.0±1.0 vs 6.7±0.8; p=0.026). The mortality rate was similar in both genotypes (p=0.255) being on average 12%, therefore at 28d after farrowing the litter size remained higher in BI sows (8.5±0.8 vs 6.1±0.6; p= 0.032). A subset of each genotype (4 gilts) was supervised during farrowing and lactation (until 28d) and piglets were weighed at birth, 24h and 28d of live. Farrowing length was not significantly different (p=0.253) between genotypes, averaging 97±22 min. When compared to ALBI (AL x BI) piglets, the BIAL (BI x AL) piglets were heavier at birth (1402±46g vs 1209±36g; p=0.002). Colostrum intake of piglets per kg of birth weight on the first 24h of life was similar between genotypes (p=0.735) being 289±15g for ALBI and 281±19g for BIAL piglets. The growth rate of piglets from birth to 28d and piglet weight at 28d was not different between genotypes (p=0.161 and p=0.091) averaging 195±6g and 6761±181g, respectively. Litter weight at 28d tended (P=0.06) to be higher on ALBI litters (56.6±4.0kg) than BIAL litters (43.2±4.0kg). This results obtained within the frame of Treasure project* are, at our knowledge, the first data of these crossbred piglets and could be used in future as reference for further studies and also for farmers that may try these cross on a commercial basis.

Newly developed electrosynthesis of Layered Double Hydroxides: application to sensing, energy storage and conversion

Layered Double hydroxides (LDHs) have been widely studied for their plethora of fascinating features and applications. The potentiostatic electrodeposition of LDHs has been extensively applied in the literature as a fast and direct method to substitute classical chemical routes. However, it does not usually allow for a fine control of the M(II)/M(III) ratio in the synthesized material and it is not suitable for large anions intercalation. Therefore, in this work a novel protocol has been proposed with the aim to overcome all these constraints using a method based on potentiodynamic synthesis. LDHs of controlled composition were prepared using different molar ratios of the trivalent to bivalent cations in the electrolytic solution ranging from 1:1 to 1:4. Moreover, we were able to produce electrochemically LDHs intercalated with carbon nanomaterials for the first time. A one-step procedure which contemporaneously allows for the Ni/Al-LDH synthesis, the reduction of graphene oxide (GO) and its intercalation inside the structure has been developed. The synthesised materials have been applied in several fields of interest. First of all, LDHs with a ratio 3:1 were exploited, and displayed good performances as catalysts for 5-(hydroxymethyl)furfural electro-oxidation, thus suggesting to carry out further investigation for applications in the field of industrial catalysis. The same materials, but with different metals ratios, were tested as catalysts for Oxygen Evolution Reaction, obtaining results comparable to LDHs synthesised by the classical co-precipitation method and also a better activity with respect to LDHs obtained by the potentiostatic approach. The composite material based on LDH and reduced graphene oxide was employed to fabricate a cathode of a hybrid supercapacitor coupled with an activated carbon anode. We can thus conclude that, to date, the potentiodynamic method has the greatest potential for the rapid synthesis of reproducible films of Co and Ni-based LDHs with controlled composition.

CFD Applications for Wave Energy Conversion Devices (MoonWEC) and Turbulent Fountains for Environmental Fluid Mechanics

This thesis is based on two studies that are related to floating wave energy conversion (WEC) devices and turbulent fountains. The ability of the open-source CFD software OpenFOAM® has been studied to simulate these phenomena. The CFD model has been compared with the physical experimental results. The first study presents a model of a WEC device, called MoonWEC, which is patented by the University of Bologna. The CFD model of the MoonWEC under the action of waves has been simulated using OpenFOAM and the results are promising. The reliability of the CFD model is confirmed by the laboratory experiments, conducted at the University of Bologna, for which a small-scale prototype of the MoonWEC was made from wood and brass. The second part of the thesis is related to the turbulent fountains which are formed when a heavier source fluid is injected upward into a lighter ambient fluid, or else a lighter source fluid is injected downward into a heavier ambient fluid. For this study, the first case is considered for laboratory experiments and the corresponding CFD model. The vertical releases of the source fluids into a quiescent, uniform ambient fluid, from a circular source, were studied with different densities in the laboratory experiments, conducted at the University of Parma. The CFD model has been set up for these experiments. Favourable results have been observed from the OpenFOAM simulations for the turbulent fountains as well, indicating that it can be a reliable tool for the simulation of such phenomena.

The FOOT experiment: Trigger and Data Acquisition (TDAQ) development and data analysis

Hadrontherapy employs high-energy beams of charged particles (protons and heavier ions) to treat deep-seated tumours: these particles have a favourable depth-dose distribution in tissue characterized by a low dose in the entrance channel and a sharp maximum (Bragg peak) near the end of their path. In these treatments nuclear interactions have to be considered: beam particles can fragment in the human body releasing a non-zero dose beyond the Bragg peak while fragments of human body nuclei can modify the dose released in healthy tissues. These effects are still in question given the lack of interesting cross sections data. Also space radioprotection can profit by fragmentation cross section measurements: the interest in long-term manned space missions beyond Low Earth Orbit is growing in these years but it has to cope with major health risks due to space radiation. To this end, risk models are under study: however, huge gaps in fragmentation cross sections data are currently present preventing an accurate benchmark of deterministic and Monte Carlo codes. To fill these gaps in data, the FOOT (FragmentatiOn Of Target) experiment was proposed. It is composed by two independent and complementary setups, an Emulsion Cloud Chamber and an electronic setup composed by several subdetectors providing redundant measurements of kinematic properties of fragments produced in nuclear interactions between a beam and a target. FOOT aims to measure double differential cross sections both in angle and kinetic energy which is the most complete information to address existing questions. In this Ph.D. thesis, the development of the Trigger and Data Acquisition system for the FOOT electronic setup and a first analysis of 400 MeV/u 16O beam on Carbon target data acquired in July 2021 at GSI (Darmstadt, Germany) are presented. When possible, a comparison with other available measurements is also reported.

Mitigation of Train-Induced waves by resonant filled-trench (RFT)

The increase of railways near the urban areas is a significant cause of discomfort for inhabitants due to train-induced vibration and noise. Vibration characteristics can vary widely according to the train type: for high-speed trains, if train speed becomes comparable to the ground wave speed, the vibration level becomes significant; for freight trains, due to their heavier weight and lower speed, the vibration amplitudes are greater and propagate at a more considerable distance from the track; for urban tramways, although the vibration amplitude is relatively low, they can have a negative structural effect on the closest buildings [51]. Therefore, to dampen the vibration level, it is possible to carry out some interventions both on the track and the transmission path. This thesis aims to propose and numerically investigate a novel method to dampen the train-induced vibrations along the transmission path. The method is called "resonant filled-trench (RFT)" and consists of a combination of expanded polystyrene (EPS) geofoam to stabilize the trench wall against the collapse and drowned cylindrical embedded inclusions inside the geofoam, which act as a resonator, reflector, and attenuator. By means of finite element simulations, we show that up to 50% higher attenuation than the open trench is achievable after overcoming the resonance frequency of the inclusion, i.e., 35Hz, which covers the frequency contents of the train-induced vibration. Moreover, depending on the filling material used for the inclusions, trench depth can be reduced up to 17% compared to the open trench showing the same screening performance as the open trench. Also, an RFT with DS inclusion installed in dense sand soil shows a high hindrance performance (i.e., IL≥6dB) when the trench depth is larger than 0.5λ_R while it is 0.6λ_R for the open trench.

Many-body approach to the BCS-BEC crossover for an imbalanced ultra-cold Fermi gas

The aim of this thesis is the study of the normal phase of a mass imbalanced and polarized ultra-cold Fermi gas in the context of the BCS-BEC crossover, using a diagrammatic approach known as t-matrix approximation. More specifically, the calculations are implemented using the fully self-consistent t-matrix (or Luttinger- Ward) approach, which is already experimentally and numerically validated for the balanced case. An imbalance (polarization) between the two spin populations works against pairing and superfluidity. For sufficiently large polarization (and not too strong attraction) the system remains in the normal phase even at zero temperature. This phase is expected to be well described by the Landau’s Fermi liquid theory. By reducing the spin polarization, a critical imbalance is reached where a quantum phase transition towards a superfluid phase occurs and the Fermi liquid description breaks down. Depending on the strength of the interaction, the exotic superfluid phase at the quantum critical point (QCP) can be either a FFLO phase (Fulde-Ferrell-Larkin-Ovchinnikov) or a Sarma phase. In this regard, the presence of mass imbalance can strongly influence the nature of the QCP, by favouring one of these two exotic types of pairing over the other, depending on whether the majority of the two species is heavier or lighter than the minority. The analysis of the system is made by focusing on the temperature-coupling-polarization phase diagram for different mass ratios of the two components and on the study of different thermodynamic quantities at finite temperature. The evolution towards a non-Fermi liquid behavior at the QCP is investigated by calculating the fermionic quasi-particle residues, the effective masses and the self-energies at zero temperature.