Using ASELL as a framework for driving change

A 2009 Australian Council of Deans of Science report (Rice, Thomas and O'Toole, 2009) identified the quality of undergraduate laboratory curricula as an important issue in tertiary science education. Evaluation of undergraduate labs traditionally relies on anecdotal evidence about what works and why; changes are often ad-hoc. We need an appropriate evidence-based methodology to clearly articulate the goals and aims of the teaching labs, and evaluate the curricula against these. Fortunately, such a methodology exists: the ASELL framework (ASELL, 2012) employs research-led surveys and workshops to identify pedagogical and logistical issues with science laboratory experiments, and an iterative process for improvement. Building on past experience with this framework, we are using the ASELL tools to examine laboratory experiments in the Schools of Physics and Molecular Bioscience at the University of Sydney to gain feedback about the level, relevance, degree of challenge, experimental techniques and class logistics. We will identify several experiments at the first year (physics) and second year (biochemistry) level to focus on, and implement changes based on the ASELL analysis — and so armed, to influence colleagues to spread the framework across the Faculty. J.W. Rice, S.M. Thomas, P. O'Toole, Tertiary Science Education in the 21st Century, Australian Learning & Teaching Council, 2009. ASELL, Advancing Science by Enhancing Learning in the Laborotory (2012), http://www.asell.or

SpoT Induces Intracellular Salmonella Virulence Programs in the Phagosome.

Guanosine tetraphosphate (ppGpp) and guanosine pentaphosphate (pppGpp), together named (p)ppGpp, regulate diverse aspects of Salmonella pathogenesis, including synthesis of nutrients, resistance to inflammatory mediators, and expression of secretion systems. In Salmonella, these nucleotide alarmones are generated by the synthetase activities of RelA and SpoT proteins. In addition, the (p)ppGpp hydrolase activity of the bifunctional SpoT protein is essential to preserve cell viability. The contribution of SpoT to physiology and pathogenesis has proven elusive in organisms such as Salmonella, because the hydrolytic activity of this RelA and SpoT homologue (RSH) is vital to prevent inhibitory effects of (p)ppGpp produced by a functional RelA. Here, we describe the biochemical and functional characterization of a spoT-Δctd mutant Salmonella strain encoding a SpoT protein that lacks the C-terminal regulatory elements collectively referred to as "ctd." Salmonella expressing the spoT-Δctd variant hydrolyzes (p)ppGpp with similar kinetics to those of wild-type bacteria, but it is defective at synthesizing (p)ppGpp in response to acidic pH. Salmonella spoT-Δctd mutants have virtually normal adaptations to nutritional, nitrosative, and oxidative stresses, but poorly induce metal cation uptake systems and Salmonella pathogenicity island 2 (SPI-2) genes in response to the acidic pH of the phagosome. Importantly, spoT-Δctd mutant Salmonella replicates poorly intracellularly and is attenuated in a murine model of acute salmonellosis. Collectively, these investigations indicate that (p)ppGpp synthesized by SpoT serves a unique function in the adaptation of Salmonella to the intracellular environment of host phagocytes that cannot be compensated by the presence of a functional RelA.IMPORTANCE Pathogenic bacteria experience nutritional challenges during colonization and infection of mammalian hosts. Binding of the alarmone nucleotide guanosine tetraphosphate (ppGpp) to RNA polymerase coordinates metabolic adaptations and virulence gene transcription, increasing the fitness of diverse Gram-positive and Gram-negative bacteria as well as that of actinomycetes. Gammaproteobacteria such as Salmonella synthesize ppGpp by the combined activities of the closely related RelA and SpoT synthetases. Due to its profound inhibitory effects on growth, ppGpp must be removed; in Salmonella, this process is catalyzed by the vital hydrolytic activity of the bifunctional SpoT protein. Because SpoT hydrolase activity is essential in cells expressing a functional RelA, we have a very limited understanding of unique roles these two synthetases may assume during interactions of bacterial pathogens with their hosts. We describe here a SpoT truncation mutant that lacks ppGpp synthetase activity and all C-terminal regulatory domains but retains excellent hydrolase activity. Our studies of this mutant reveal that SpoT uniquely senses the acidification of phagosomes, inducing virulence programs that increase Salmonella fitness in an acute model of infection. Our investigations indicate that the coexistence of RelA/SpoT homologues in a bacterial cell is driven by the need to mount a stringent response to a myriad of physiological and host-specific signatures

Comparative analysis of intrathecal dexmedetomidine and fentanyl as an adjuvant to isobaric levobupivacaine in orthopaedic surgery

Aim: to compare a combination of isobaric Levobupivacaine with fentanyl and dexmedetomidine for the characteristics of spinal blockade with respect to onset, duration and hemodynamic parameters and side effect. Material and methods: This was a prospective, randomized, and double blinded clinical comparative study conducted in the Department of Anaesthesiology, Jawaharlal Nehru medical college and hospital Bhagalpur, Bihar, India from march 2019 to February 2020. The study population consisted of 150 adult patients who were classified as American Society of Anesthesiologists (ASA) physical status I or II, undergoing elective lower limb orthopaedic surgery under spinal anesthesia. The study participants were randomly divided into three groups.Group A: 0.5% Levobupivacaine Isobaric 2.5ml+ 0.5ml normal saline (total volume is upto 3.0 ml). Group F: 0.5% Levobupivacaine Isobaric 2.5ml + 25mug fentanyl (test solution will diluted with normal saline to total volume of 3.0ml). Group D: 0.5% Levobupivacaine isobaric 2.5ml +5 mcg dexmedetomidine (test solution will diluted with normal saline to total volume of 3.0 ml. Result: The mean time for onset of sensory block was 10.59 ±4.06 min in the saline group and 8.31±2.93 min in the dexmedetomidine group  and 2.21±1.19 min in the fentanyl group. The mean time taken to achieve maximum sensory block in group A was 15.62±4.91 min, in group D was 13.18±3.51 min and in group F it was 5.28±1.92 min so maximum sensory block was achieved earlier in group. Peak level of sensory block attained in the fentanyl group was T4 and the peak level of sensory block in dexmedetomidine group was T6 and in the saline group peak level was T8. So the highest sensory block was attained in the fentanyl group. The mean duration of sensory block in group A was 113.47±7.12min, and in group F was 161.11±12.74min., and in group D was 204.12±6.41 min. Prolong duration occur in the dexmedetomiine group. The prolongation of effect may result from synergism between local anaesthetic and alpha2 adrenoceptor agonist action. The mean onset time of motor block in group A was 11.04±3.98 min, in group D it was 8.97±3.31 min, in group F it was 3.31 ±1.33 min. Onset of motor block occured earlier in the fentanyl group. In the present study there was a significant difference in duration of motor block across the three groups with p value <0.001. In group A mean duration of motor block was 161.04±6.30 min, and in group D was 253.26±6.59 min and in group F it was 186.74±11.64 min. Conclusion:Dexmedetomidine group has longer onset of and duration of sensory block and effective postoperative analgesia and fewer side effect as compared to fentanyl group. Keywords: Subarachanoid block, Levobupivacaine, anesthesia

The methylglyoxal pathway is a sink for glutathione in Salmonella experiencing oxidative stress.

Salmonella suffer the cytotoxicity of reactive oxygen species generated by the phagocyte NADPH oxidase in the innate host response. Periplasmic superoxide dismutases, catalases and hydroperoxidases detoxify superoxide and hydrogen peroxide (H2O2) synthesized in the respiratory burst of phagocytic cells. Glutathione also helps Salmonella combat the phagocyte NADPH oxidase; however, the molecular mechanisms by which this low-molecular-weight thiol promotes resistance of Salmonella to oxidative stress are currently unknown. We report herein that Salmonella undergoing oxidative stress transcriptionally and functionally activate the methylglyoxal pathway that branches off from glycolysis. Activation of the methylglyoxal pathway consumes a substantial proportion of the glutathione reducing power in Salmonella following exposure to H2O2. The methylglyoxal pathway enables Salmonella to balance glucose utilization with aerobic respiratory outputs. Salmonella take advantage of the metabolic flexibility associated with the glutathione-consuming methylglyoxal pathway to resist reactive oxygen species generated by the enzymatic activity of the phagocyte NADPH oxidase in macrophages and mice. Taken together, glutathione fosters oxidative stress resistance in Salmonella against the antimicrobial actions of the phagocyte NADPH oxidase by promoting the methylglyoxal pathway, an offshoot metabolic adaptation of glycolysis

A study to assess the antimicrobial resistance pattern of bacteria isolated from ICU patients with respiratory tract infections

Aim: The aim of this study to analyzed antimicrobial Resistance Pattern of Bacteria Isolated from ICU Patients with Respiratory Tract Infections. Materials and Methods: This was a prospective, observational study conducted in the Department of Anaesthesiology, Jawaharlal Nehru medical college and hospital Bhagalpur, Bihar, India from January 2019 to December 2019. A total of 200 samples (Sputum, Pleural fluid, Pus, IT-Tube sample, Bronchial swab, Respiratory swab) of RTI patients were collected from from Respiratory ICU. For identification of different microorganisms selective media were used in this study. Isolate the microorganisms with the help of primary and secondary identification and antibiotic sensitivity were performed. Results:Majority of the patients was male 62.5% and rest 37.5% was female. Gram negative organisms are detected in (67%), Gram positive organisms detected (22%), and no organism detected (11%) of the swab samples. Conclusion: Piperacillin-tazobactam was the most common antibiotic prescribed to patients with respiratory infection admitted to ICU. More than half of patients had resistance to the empirical antibiotic used in our ICU, highlighting the need for antibiogram for each ICU. Most of the patient had prior antibiotic use and had mainly gram negative organisms with high resistance to commonly used antibiotics

Identification of a Catabolite-Responsive Element Necessary for Regulation of the cry4A Gene of Bacillus thuringiensis subsp. israelensis▿ †

Bacillus thuringiensis subsp. israelensis produces a potent mosquitocidal protein, Cry4A. We have identified a 15-bp catabolite responsive element (cre), overlapping the −35 element of the cry4A promoter. Changing a guanine to adenine at position −49 in the promoter abolished glucose catabolite repression of cry4A and enhanced promoter activity two- to threefold. This cis regulatory element is essential for controlled toxin synthesis, vital to evolutionary success of B. thuringiensis subsp. israelensis

Role of non-invasive ventilation (NIV) for treatment of acute respiratory failure (ARF) among patients with non cystic fibrosis (CF) bronchiectasis

Aim: The aim of this study to evaluate the role of non-invasive ventilation (NIV) for treatment of acute respiratory failure (ARF) among patients with non cystic fibrosis (CF) bronchiectasis. Material and methods: This was a retrospective study conducted in the Department of Anaesthesiology, Jawaharlal Nehru medical college and hospital Bhagalpur, Bihar, India from May 2018 to February 2019. We included 100 patients with bronchiectasis and ARF who required either NIV or invasive mechanical ventilation (IMV). Results:  There was a total of 200 patients with bronchiectasis, Among these, 100 patients were admitted with ARF. Totally 100 patients who required either NIV or IMV. The most common etiology of bronchiectasis was post‑tuberculosis (50%) followed by idiopathic (20%), ABPA (15%), and immunodeficiency (5%). NIV was initiated as first line of ventilator support for 80 patients. Among these, 51(63.75%) were managed successfully with NIV. 29 (36.25%) patients failed NIV and required endotracheal intubation during the hospital stay. Reasons for NIV failure were worsening or non-improvement of ventilatory or oxygenation parameters (n=14), hypotension (n=6), worsening of sensorium (n=4), and intolerance (n=5). NIV failure occurred after a median duration of 2.77(95% confidence interval [CI] 1.51–4.24) days after the initiation. There were total 11 deaths in the study group. Among patients who failed NIV, total days (median [range]) spent on ventilator (6.6 [2–62] vs. 6.1 [3–16] days; P=0.41), duration (median [range]) of hospital stay (8 [4–64] vs. 11 [5–15] days; P=0.27), and mortality (8 [10%] vs 3 [15%]; P=0.24) were comparable to the IMV group. The causes of death among patients who failed NIV were septic shock (n=5) and ventilator-associated pneumonia (n=3). Conclusions: NIV is feasible for management of ARF with non-CF bronchiectasis. High APACHE may predict NIV failure among these patients. Keywords: Acute respiratory failure, mechanical ventilation, noncystic fibrosis bronchiectasis, noninvasive ventilation

A Study of Knowledge regarding Health Impact of Tobacco and Attitude towards Quitting among Lower-Middle and Lower Classes in a Resettlement Colony in Delhi, India

Objectives: To study knowledge regarding health impact of tobacco and attitude towards quitting among lower-middle and lower classes in a resettlement colony in Delhi.Materials and Methods: The study was conducted between the months of March and October in the year 2011, in a resettlement colony of Delhi. The study was a cross-sectional, community-based, house-to-house survey and involved interviews of present tobacco consumers using a structured interview guide consisting mainly of close-ended questions and some open-ended questions.Results: 1698 subjects were enrolled in the study with 90.8% males and 8.7% females. Mean age of the subjects was 40 years, while median age was 37 years. 87% of the respondents were educated up to higher secondary or less. Some other key findings included the following:45% of the respondents had no information on the effect of tobacco consumption on health, and 60% did not have any knowledge of the laws regarding tobacco consumption. Television was the most common source of information regarding tobacco consumption, followed by newspapers. Three-fourths of the respondents had knowledge that their using tobacco had an impact on other family members, and 60% believed that children were affected most commonly.Spouse or children were the most common people who stressed upon the respondents to leave tobacco, followed by parents. Self-motivation was the most commonly reported second reason for attempting to quit tobacco. However, doctors or other medical personnel appeared to play only a marginal role in motivating the tobacco users to quit tobacco.Conclusion: This study shows that there is tremendous scope for improving the outcomes in tobacco control by strengthening the programmatic components of tobacco control such as propagating the health effects of tobacco in a manner intelligible to the people and making the health system to proactively contribute in anti-tobacco efforts

Duality in vector optimization in Banach spaces with generalized convexity

We consider a vector optimization problem with functions defined on Banach spaces. A few sufficient optimality conditions are given and some results on duality are proved

<i>M.tb-Rv2462c</i> of <i>Mycobacterium tuberculosis</i> Shows Chaperone-like Activity and Plays a Role in Stress Adaptation and Immunomodulation

Mycobacterium tuberculosis (M.tb)-encoded factors protect it against host-generated stresses and support its survival in the hostile host environment. M.tb possesses two peptidyl-prolyl cis-trans isomerases and a probable trigger factor encoded by Rv2462c which has an FKBP-like PPIase domain. PPIases are known to assist the folding of peptidyl-prolyl bonds and are involved in various cellular processes important for bacterial survival in host-generated stresses. In this study, we aim to functionally characterize Rv2462c of M.tb. Our data suggest that the trigger factor of M.tb exhibits chaperone activity both in vitro and in vivo. Heterologous expression of M.tb-Rv2462c locus into Mycobacterium smegmatis enhanced its survival within macrophages, adaptation to oxidative stress and biofilm formation. M.tb-trigger factor has strong immunomodulatory potential and modifies the cytokine profile of the host towards the proinflammatory axis