Experimental study on mixing of activated sludge with or without air in a mixing tank through electrical resistance tomography (ERT) and response surface methodology

In such industrial processes as oxidation, hydrogenation, and biological fermentation gas and liquid are contacted and mixed to reach steady condition. The unit operations applied for gas/liquid processes include but are not limited to bubble column, plate column, mechanically agitated vessels, in-line static mixers, jet mixing devices/jet mixers, and surface aerators. Among aforementioned unit operations, aerated mixing vessels are mostly employed for the gas-liquid processes in such industry as biochemical, pharmaceutical, cosmetics, and waste water treatment. While gas-dispersion in Newtonian fluids, especially in low-viscosity systems, has been successfully understood, not enough information may be found in the literature about this process in non-Newtonian fluids. Therefore, in this study electrical resistance tomography (ERT) was utilized to assess the mixing of the activated sludge as shear thinning non-Newtonian fluid in presence of aeration. ERT results revealed that shorter mixing time can be achieved in presence of aeration. The following three central impellers were employed: ASI (a combination of A200 and the Scaba impellers), ARI (a combination of A200 and the Rushton impellers), and Rushton (fully radial impeller). An ERT system with a two-plane assembly equipped with 16 sensors on each plane was employed to assess the impact of the impeller type, impeller speed, and gas flow rate on the mixing of activated sludge in terms of mixing time, specific power consumption, and gas flow number. A statistical-based experimental design with RSM (response surface methodology) was applied to evaluate the individual and interactive effects of the design parameters and operating conditions. Experiments and RSM demonstrated that among all independent variables in this study, impeller speed was the common independent variable which impacts mixing time, specific power consumption and gas flow number significantly

MicroRNAs in the regulation of alternatively activated macrophages

Macrophages play a key role in maintaining the balance and efficiency of the immune response. TH2 cytokines IL-4 & IL-13, through shared IL-4Rα signalling, trigger a state of alternative activation in macrophages and also drive their proliferation. Alternatively activated macrophages (AAMΦ) are involved in the control of helminth infections and have also been implicated in tissue repair. However, TH2 weighted imbalance can result in inflammatory disorders such as asthma and fibrosis. Hence, macrophage responses must be tightly regulated. MicroRNAs, a short (~22nt) class of non-coding RNA, are one such immunomodulatory feedback mechanism that can regulate gene expression by targeting the 3’ UTR of mRNA resulting in destabilisation of the mRNA and/or inhibition of translation. With their ability for vast gene regulation, it was hypothesised that microRNAs could play a crucial role in the regulation of AAMΦ by targeting genes and pathways critical for their induction, maintenance & proliferation. Previously generated microarrays in the lab have allowed us to identify microRNAs differentially expressed in AAMΦ. In an effort to determine which microRNAs are genuinely associated with alternative activation, the first part of this project examined the expression profiles of ten shortlisted microRNA candidates under varying conditions of alternative activation, ranging from a reductionist in vitro IL-4/13 stimulation of macrophage cell lines to a complex in vivo TH2 mouse model of filarial infection. Profiling of microRNA expression under these conditions revealed that the expression of two IL-4Rα dependent microRNAs, namely miR-199b-5p and miR-378, along with another microRNA, miR-146, was highly regulated and consistently associated with alternative activation. The subsequent chapters of this thesis investigated the contribution of these microRNAs in regulating AAMΦ responses. Interestingly, we identified miR-199b-5p as being highly expressed in AAMΦ in vivo but not in vitro. Pathway analysis identified insulin signalling and other proliferative pathways such as PI3K/AKT as being highly targeted by miR-199b-5p. Overexpression of miR-199b-5p in RAW 264.7 cells resulted in a reduction in the rate of proliferation and a change in the levels of Insulin Receptor Substrate -1 (IRS-1), suggesting that miR-199b- 5p might regulate macrophage proliferation via insulin signalling. An alteration in the expression of YM-1 and RELM-α, markers characteristic of alternative activation, was also observed. MiR-199b-5p was successfully delivered to the lung and overexpressed in alternatively activated alveolar macrophages. No effect was observed on IL-4 induced proliferation, potentially due to the lack of significant insulin receptor and IRS-1 expression in alveolar macrophages. However, secreted levels of YM-1, but not RELM-α, were significantly reduced. MiR-378 is a microRNA that has previously been shown to be associated with AAMΦ through targeting of AKT-1; however, a direct influence of this microRNA on the regulation of this phenotype is yet to be determined. In this thesis, we have provided direct evidence of the impact of miR-378 deficiency on the regulation of AAMΦ and their responses using miR-378 KO mice. The ability of macrophages isolated from WT and KO animals to alternatively activate was studied in various systems both in vitro and in vivo. The influence of miR-378 deficiency on IL-4 induced proliferation was also addressed in vivo. Although the lack of miR-378 had no significant effect on IL-4 driven macrophage proliferation, results from this chapter support a role for miR-378 in the regulation of alternative activation through regulation of YM-1 and RELM-α expression. Lastly, to determine whether this regulation by miR-378 had functional consequences, we also utilised Litomosoides sigmodontis, a murine model of filarial infection. Due to experimental limitations, a concrete role for miR-378 in the context of infection could not be established. The final chapter of this thesis focuses on examining the role of miR-146 in the regulation of AAMΦ. MiR-146a is a highly studied microRNA that has previously been linked strongly to TH1 immune responses, especially classical activation of macrophages. However, a role for this microRNA in regulating AAMΦ is yet to be determined. Expression levels of miR-146a and miR-146b, the two isoforms of miR-146, were found to be differentially regulated upon alternative activation, with a decrease in miR-146a and increase in miR-146b expression in response to IL-4 both in vitro and in vivo. Based on this difference in expression and their known functions in suppressing excessive proinflammatory responses, it was hypothesised that miR-146a/b serve to regulate proinflammatory molecules (and signals) in a fine balance to allow efficient alternative activation to occur. However, the high sequence similarity between these two isoforms proved to be a hindrance to test this hypothesis in terms of shared targets. Therefore, the latter half of this chapter was devoted to the generation and optimisation of a stable cell line for the identification of microRNA targets using CLASH (cross-linking, ligation and sequencing of hybrids). In summary, the results from this thesis provide an important foundation for further studies of the functional role of microRNAs in the regulation of AAMΦ. Firstly, it characterises the expression profiles of ten different microRNAs differentially expressed during alternative activation. Secondly, for the first time, it identifies a role for miR-199b- 5p in the regulation of macrophage proliferation and activation. Thirdly, this thesis has provided direct evidence for the effect of miR-378 deficiency on AAMΦ responses. Lastly, it identifies and demonstrates the robust differential expression of two separate isoforms of the same microRNA (miR-146) under varying conditions of alternative activation, whose functional properties as regulators of the AAMΦ phenotype await further investigation

Effect of oral L- arginine versus intravenous hydration on maternal and fetal outcome in idiopathic oligohydramnios

Background: Adequate amount of amniotic fluid was required for normal growth of fetus. Oligohydramnios or reduced amount of amniotic fluid is associated with adverse maternal and perinatal outcome due to increase in induced labour and operative deliveries. Idiopathic oligohydramnios is a condition in which no other risk factors are associated with pregnancy. This study was done to compare the effect of L-arginine and IV hydration on improvement of amniotic fluid index and fetal growth.Methods: Total 50 patients were included in the study according to inclusion criteria and divided equally into two groups randomly. IV hydration was given to one group and other group received L- arginine sachet orally. The effect on AFI and fetal outcome was compared.Result: The result was compared with respect to age, gravidity, gestational age and AFI at the time of study and after giving treatment. Maternal and fetal outcome was compared which shows that L-arginine was more effective in increasing the AFI and thereby leading to favorable results in the form of increase in gestational age at time of delivery and fetal weight.Conclusion: This study shows that both IV hydration and L-arginine are useful in treatment of oligohydramnios. But L-arginine appears more advantageous over IV hydration in improving pregnancy outcome and reducing perinatal morbidity and mortality

A Review of Data Relevant to Agriculture in India

The Government of India, through more than half a dozen ministries and various research institutions, regularly collects and reports data relevant to the understanding of food production across the country. The aim of this review was to summarize these data sources. Results indicate that while many datasets are publicly available, there is little coordination between ministries. This has resulted in replication of efforts and inconsistencies in reported values for key indicators. Various other factors, including, for example, reporting in PDF format, frequent changes in district boundaries, missing data, and time lags in reporting, further hinder utilization of these data. A standardized data collection protocol for timely reporting of high-quality, complete data is needed. Such a protocol could be developed and overseen by a central, inter-ministerial Agri-Food System Data Governance Steering Committee. The establishment of such a committee would help ensure data-driven, evidence-based policymaking towards a nutrition secure India

Genetic Variation Is the Major Determinant of Individual Differences in Leukocyte Endothelial Adhesion

Objective: To determine the genetic contribution to leukocyte endothelial adhesion.Methods: Leukocyte endothelial adhesion was assessed through a novel cell-based assay using human lymphoblastoid cell lines. A high-throughput screening method was developed to evaluate the inter-individual variability in leukocyte endothelial adhesion using lymphoblastoid cell lines derived from different donors. To assess heritability, ninety-two lymphoblastoid cell lines derived from twenty-three monozygotic twin pairs and twenty-three sibling pairs were compared. These lymphoblastoid cell lines were plated with the endothelial cell line EA.hy926 and labeled with Calcein AM dye. Fluorescence was assessed to determine endothelial cell adhesion to each lymphoblastoid cell line. Intra-pair similarity was determined for monozygotic twins and siblings using Pearson pairwise correlation coefficients.Results: A leukocyte endothelial adhesion assay for lymphoblastoid cell lines was developed and optimized (CV = 8.68, Z′-factor = 0.67, SNR = 18.41). A higher adhesion correlation was found between the twins than that between the siblings. Intra-pair similarity for leukocyte endothelial adhesion in monozygotic twins was 0.60 compared to 0.25 in the siblings. The extent to which these differences are attributable to underlying genetic factors was quantified and the heritability of leukocyte endothelial adhesion was calculated to be 69.66% (p-valueConclusions: There is a heritable component to leukocyte endothelial adhesion. Underlying genetic predisposition plays a significant role in inter-individual variability of leukocyte endothelial adhesion.</p

Young people’s experiences of COVID-19 messaging at the start of the UK lockdown:Lessons for positive engagement and information sharing

BACKGROUND: To reduce COVID-19 infection rates during the initial stages of the pandemic, the UK Government mandated a strict period of restriction on freedom of movement or 'lockdown'. For young people, closure of schools and higher education institutions and social distancing rules may have been particularly challenging, coming at a critical time in their lives for social and emotional development. This study explored young people's experiences of the UK Government's initial response to the pandemic and related government messaging.METHODS: This qualitative study combines data from research groups at the University of Southampton, University of Edinburgh and University College London. Thirty-six online focus group discussions (FGDs) were conducted with 150 young people (Southampton: n = 69; FGD = 7; Edinburgh: n = 41; FGD = 5; UCL: n = 40; FGD = 24). Thematic analysis was conducted to explore how young people viewed the government's response and messaging and to develop recommendations for how to best involve young people in addressing similar crises in the future. RESULTS: The abrupt onset of lockdown left young people shocked, confused and feeling ignored by government and media messaging. Despite this, they were motivated to adhere to government advice by the hope that life might soon return to normal. They felt a responsibility to help with the pandemic response, and wanted to be productive with their time, but saw few opportunities to volunteer.CONCLUSIONS: Young people want to be listened to and feel they have a part to play in responding to a national crisis such as the COVID-19 epidemic. To reduce the likelihood of disenfranchising the next generation, Government and the media should focus on developing messaging that reflects young people's values and concerns and to provide opportunities for young people to become involved in responses to future crises

Effects of antiplatelet therapy on stroke risk by brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases: subgroup analyses of the RESTART randomised, open-label trial

Background Findings from the RESTART trial suggest that starting antiplatelet therapy might reduce the risk of recurrent symptomatic intracerebral haemorrhage compared with avoiding antiplatelet therapy. Brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases (such as cerebral microbleeds) are associated with greater risks of recurrent intracerebral haemorrhage. We did subgroup analyses of the RESTART trial to explore whether these brain imaging features modify the effects of antiplatelet therapy

Daidzein Prevents the Increase in CD4+CD28null T Cells and B Lymphopoesis in Ovariectomized Mice: A Key Mechanism for Anti-Osteoclastogenic Effect

Estrogen deficiency leads to an upregulation of TNF-α producing T cells and B-lymphopoesis which augments osteoclastogenesis. Estrogen deficiency also increases the population of premature senescent CD4+CD28null T cells which secrete a higher amount of TNF-α thus leading to enhanced osteoclastogenesis. Isoflavonoids like daidzein and genistein are found mostly in soybeans, legumes, and peas. These share structural similarity with 17β-stradiol (E2) and have osteoprotective role. This study explores the effect of daidzein (Daid) on the proliferation of TNF-α producing T cells, premature senescent T cells and B cell lymphopoesis under estrogen deficient conditions. For this study adult Balb/c mice were treated with Daid at 10 mg/kg body weight dose by oral gavage daily post ovariectomy (Ovx). After six weeks animals were autopsied and bone marrow and spleen cells were collected for FACS analysis. Blood serum was collected for ELISA. It was observed that Ovx mice treated with Daid for six weeks show reduction in Ovx induced expansion of CD4+ T cells in bone marrow and spleen when analysed by flow cytometry. Estrogen deficiency led to increased prevalence of TNF-α secreting CD4+CD28null T cells, however, treatment with Daid increased the percentage of CD4+CD28+ T cells. Co-culture of CD4+CD28null T cells and bone marrow resulted in enhanced osteoclastogenesis as evident by increased tartarate resistant acid phosphatase (TRAP) expression, an osteoclast marker. However, treatment with Daid resulted in reduced osteoclastogenesis in CD4+CD28null T cells and bone marrow cell co-culture. Daid also regulated B lymphopoesis and decreased mRNA levels of RANKL in B220+ cells. Taken together, we propose that one of the mechanisms by which Daid prevents bone loss is by reversing the detrimental immune changes as a result of estrogen deficiency

Fast acting allosteric phosphofructokinase inhibitors block trypanosome glycolysis and cure acute African trypanosomiasis in mice

The parasitic protist Trypanosoma brucei is the causative agent of Human African Trypanosomiasis, also known as sleeping sickness. The parasite enters the blood via the bite of the tsetse fly where it is wholly reliant on glycolysis for the production of ATP. Glycolytic enzymes have been regarded as challenging drug targets because of their highly conserved active sites and phosphorylated substrates. We describe the development of novel small molecule allosteric inhibitors of trypanosome phosphofructokinase (PFK) that block the glycolytic pathway resulting in very fast parasite kill times with no inhibition of human PFKs. The compounds cross the blood brain barrier and single day oral dosing cures parasitaemia in a stage 1 animal model of human African trypanosomiasis. This study demonstrates that it is possible to target glycolysis and additionally shows how differences in allosteric mechanisms may allow the development of species-specific inhibitors to tackle a range of proliferative or infectious diseases