Involvement of a nine-residue loop of streptokinase in the generation of macromolecular substrate specificity by the activator complex through interaction with substrate kringle domains

The selective deletion of a discrete surface-exposed epitope (residues 254-262; 250-loop) in the β domain of streptokinase (SK) significantly decreased the rates of substrate human plasminogen (HPG) activation by the mutant (SKdel254-262). A kinetic analysis of SKdel254-262 revealed that its low HPG activator activity arose from a 5-6-fold increase in Km for HPG as substrate, with little alteration in kcat rates. This increase in the Km for the macromolecular substrate was proportional to a similar decrease in the binding affinity for substrate HPG as observed in a new resonant mirror-based assay for the real-time kinetic analysis of the docking of substrate HPG onto preformed binary complex. In contrast, studies on the interaction of the two proteins with microplasminogen showed no difference between the rates of activation of microplasminogen under conditions where HPG was activated differentially by nSK and SKdel254-262. The involvement of kringles was further indicated by a hypersusceptibility of the SKdel254-262. plasmin activator complex to ε-aminocaproic acid-mediated inhibition of substrate HPG activation in comparison with that of the nSK.plasmin activator complex. Further, ternary binding experiments on the resonant mirror showed that the binding affinity of kringles 1-5 of HPG to SKdel254-262.HPG was reduced by about 3-fold in comparison with that of nSK.HPG. Overall, these observations identify the 250 loop in the β domain of SK as an important structural determinant of the inordinately stringent substrate specificity of the SK.HPG activator complex and demonstrate that it promotes the binding of substrate HPG to the activator via the kringle(s) during the HPG activation process

Serological Prediction of infections in Diabetic Patients with Diabetes Ketoacidosis in Penang, Malaysia

Purpose: To determine the prevalence and predictors of infection in diabetic patients with diabetic ketoacidosis (DKA) who were .18 years.Methods: A retrospective cohort design was adopted for this study. A total of 967 diabetes ketoacidosis patients from Hospital Pulau Pinang for the 3-year period, Jan 2008 - Dec 2010, were identified and enrolled. The data were analysed, as appropriate, by Student t-test and ANOVA for the normally distributed data, Mann-Whitney U rank sum and Kruskall-Wallis tests for continuous, non-nominal data and Chi-square for dichotomous variables. Odd Ratios with 95% confidence interval (CI) were alsopresented where applicable.Results: Of the total diabetes ketoacidosis patients, 112 (11.6 %) were cases without infection, 679 (70.2 %) bacterial infection cases and 176 (18.2 %) presumed viral infection cases. The mean white blood count (WBC) for all the patients was 18,177 } 9,431 while 721 (74.6 %) had leukocytosis, defined by WBC . 15,000/mm3. WBC differential, leukocytosis, as well as sex and body temperature were not significant predictors (p >.05) of bacterial infection. There was, however, a significant difference (p <.05) in terms of age within groups, as those > 57 years showed a higher rate of infection.Conclusion: The infection rate in elderly patients with DKA is high and a majority of them lack clinical evidence. Age has a significant effect on the rate and prediction of infection. Leukocytosis is commonly found but severe ketoacidosis was more likely than the presence of infection.Keywords: Diabetes mellitus, Diabetes ketoacidosis, Infections, Predictors, White blood cell

Domain truncation studies reveal that the streptokinase-plasmin activator complex utilizes long range protein-protein interactions with macromolecular substrate to maximize catalytic turnover

To explore the interdomain co-operativity during human plasminogen (HPG) activation by streptokinase (SK), we expressed the cDNAs corresponding to each SK domain individually (α , β , and γ ), and also their two-domain combinations, viz. αβ and βγ in Escherichia coli. After purification, α and β showed activator activities of approximately 0.4 and 0.05%, respectively, as compared with that of native SK, measured in the presence of human plasmin, but the bi-domain constructs αβ and βγ showed much higher co-factor activities (3.5 and 0.7% of native SK, respectively). Resonant Mirror-based binding studies showed that the single-domain constructs had significantly lower affinities for "partner" HPG, whereas the affinities of the two-domain constructs were remarkably native-like with regards to both binary-mode as well as ternary mode ("substrate") binding with HPG, suggesting that the vast difference in co-factor activity between the two- and three-domain structures did not arise merely from affinity differences between activator species and HPG. Remarkably, when the co-factor activities of the various constructs were measured with microplasminogen, the nearly 50-fold difference in the co-factor activity between the two- and three-domain SK constructs observed with full-length HPG as substrate was found to be dramatically attenuated, with all three types of constructs now exhibiting a low activity of approximately 1-2% compared to that of SK·HPN and HPG. Thus, the docking of substrate through the catalytic domain at the active site of SK-plasmin(ogen) is capable of engendering, at best, only a minimal level of co-factor activity in SK·HPN. Therefore, apart from conferring additional substrate affinity through kringle-mediated interactions, reported earlier (Dhar et al., 2002; J. Biol. Chem. 277, 13257), selective interactions between all three domains of SK and the kringle domains of substrate vastly accelerate the plasminogen activation reaction to near native levels

Plan-do-Check-Act Cycle: a Method to Improve Customer Satisfaction at a Municipal Council in Malaysia

Purpose: This study aimed to determine the use of PDCA methods in improving customer satisfaction at SP MUNICIPAL COUNCIL. Maintaining customer satisfaction is the main responsibility of my department as we are in charge of receiving complaints from the public and providing the best solution to the public. Since the major operation for SP MUNICIPAL COUNCIL is providing services to the public, it is crucial to solving any related issue regarding poor customer services and management by SP MUNICIPAL COUNCIL, long delays in solving issues from the public and others. Hence, using appropriate tools such as the PDCA method is one of the ways to achieve a continuous improvement process and improve the service quality for SP MUNICIPAL COUNCIL. The aim of this study is to overcome this issue and improve customer satisfaction, the quality of our services and reduce complaints from the public towards the management of SP MUNICIPAL COUNCIL by using the PDCA method.   Theoretical framework: Problem identification needs to be done to find the causes of customer complaints. From the findings, the factors that potentially cause the problem are customer services, workers, administration, and management system issues. Then, the next step is a repair process where the result after improvement, implementation and evaluation will be measured.   Design/methodology/approach: The method used for this study is the PDCA method in the literature review. The data of this study are compiled, analyzed and concluded, to obtain the results for the method in the literature review. The literature review compares several research papers that implement PDCA methods.   Findings: The findings show that Plan-Do-Act-Check method can give a positive impact on increasing service quality for SP MUNICIPAL COUNCIL. There are several solution and recommendation have been achieved after the discussion of the result. These solutions will help in overcome the issue faced by SP MUNICIPAL COUNCIL.   Research limitation/implication: This research only focuses on improving customer satisfaction at SP MUNICIPAL COUNCIL and only applies to departments that focus on customer services. Originality/value: This research contributes to existing literature and use new findings to overcome problem using PDCA method and fishbone analysis to identify the cause and effect of the problem

Retrocaval ureter: report of 2 different clinical spectrums

Retrocaval ureter is a relatively rare anomaly where ureteric obstruction may occur as a result of ureter passes behind the inferior vena cava (IVC), hence, compressing it between the IVC and the vertebrae. We report 2 cases of retrocaval ureter with different presentations. One patient was managed surgically with minimally invasive approach and the other was managed conservatively

Composition and Functionality of Lipid Emulsions in Parenteral Nutrition: Examining Evidence in Clinical Applications

Lipid emulsions (LEs), an integral component in parenteral nutrition (PN) feeding, have shifted from the primary aim of delivering non-protein calories and essential fatty acids to defined therapeutic outcomes such as reducing inflammation, and improving metabolic and clinical outcomes. Use of LEs in PN for surgical and critically ill patients is particularly well established, and there is enough literature assigning therapeutic and adverse effects to specific LEs. This narrative review contrarily puts into perspective the fatty acid compositional (FAC) nature of LE formulations, and discusses clinical applications and outcomes according to the biological function and structural functionality of fatty acids and co-factors such as phytosterols, α-tocopherol, emulsifiers and vitamin K. In addition to soybean oil-based LEs, this review covers clinical studies using the alternate LEs that incorporates physical mixtures combining medium- and long-chain triglycerides or structured triglycerides or the unusual olive oil or fish oil. The Jaded score was applied to assess the quality of these studies, and we report outcomes categorized as per immuno-inflammatory, nutritional, clinical, and cellular level FAC changes. It appears that the FAC nature of LEs is the primary determinant of desired clinical outcomes, and we conclude that one type of LE alone cannot be uniformly applied to patient care

De novo transcriptome dataset of Stevia rebaudiana accession MS007

Stevia rebaudiana (S. rebaudiana) is a herbaceous and perennial plant belonging to Asteraceae family. The genus stevia is well known as a natural producer of sweetener comprising non-caloric and non-carcinogenic steviol glycosides. In recent years, the capability in producing natural sweetner has increased the demand for S. rebaudiana as substitute of processed sugars. Flowering phase of S. rebaudiana has shown to affect the content of steviol glycosides in the leaves. Steviol glycosides level is the highest at the time of flower bud formation and lowest at time preceding and following flower bud formation. Therefore, sequencing and analysing the genes that are involved in flowering phase will provide platform for gene manipulation in increasing steviol glycosides content. The Stevia transcriptome data that include two stages of growth (before flowering and after flowering), were obtained using Illumina RNA-seq technology and can be accessed at NCBI Sequence Read Archive under Accession No. SRX6362785 and SRX6362784

Nickel nanowire: magnetic ordering synthesis

Magnetic nanowires have been material of interest among researchers due to their unique magnetic properties. In the present research, Nickel (Ni) nanowires with an average diameter of 250 nm and length up to 25 μm have been successfully prepared via anodic alumina oxide (AAO) template-assisted electrodeposition method at the different magnetic field intensities and current density. The primary interest is to investigate the effect of the external magnetic field and current density on the morphological, growth length, crystal orientation and growth of the Ni nanowires. Investigation finding reveals that the employed magnetic field and current density smoothened the surface texture, improved growth length and reduced the crystal size. The observed changes are believed to be contributed by the interaction forces induced by the intensity of applied electric field and the external magnetic field known as magnetohydrodynamic (MHD) effect