Site Saturation Mutagenesis Applications on <i>Candida methylica</i> Formate Dehydrogenase

In NADH regeneration, Candida methylica formate dehydrogenase (cmFDH) is a highly significant enzyme in pharmaceutical industry. In this work, site saturation mutagenesis (SSM) which is a combination of both rational design and directed evolution approaches is applied to alter the coenzyme specificity of NAD+-dependent cmFDH from NAD+ to NADP+ and increase its thermostability. For this aim, two separate libraries are constructed for screening a change in coenzyme specificity and an increase in thermostability. To alter the coenzyme specificity, in the coenzyme binding domain, positions at 195, 196, and 197 are subjected to two rounds of SSM and screening which enabled the identification of two double mutants D195S/Q197T and D195S/Y196L. These mutants increase the overall catalytic efficiency of NAD+ to 5.6×104-fold and 5×104-fold value, respectively. To increase the thermostability of cmFDH, the conserved residue at position 1 in the catalytic domain of cmFDH is subjected to SSM. The thermodynamic and kinetic results suggest that 8 mutations on the first residue can be tolerated. Among all mutants, M1L has the best residual activity after incubation at 60°C with 17%. These studies emphasize that SSM is an efficient method for creating “smarter libraries” for improving the properties of cmFDH

The effect of ethanolic extract of premature Musa Paradisiaca (plantain) pulp on the histology of the liver and kidneys of female Wistar rats

Background: Premature plantain is a major component in herbal remedies used for the treatment of different ailment such as reducing blood sugar, and peptic ulcer disease. The aim of the study was to determine the effect of ethanolic extract of premature Musa paradisiaca on histology of the liver and kidneys of female Wistar rats.   Methods: Twenty female Wistar rats weighing between 180-200 g were divided into four groups. Group 1 was administered distilled water only, while groups 2, 3 and 4 were administered the ethanolic extract of premature Musa paradisiaca in low, medium, and high dose respectively for 14 days. Results: Twenty-four hours after the last administration, all animals were sacrificed, tissues were harvested. The histological reports showed varying level of damage to the cytoarchitecture of the liver and kidney tissues of the treatment groups when compared to the control. Conclusions: This plant may likely induce nephrotoxic and hepatotoxic changes

Efficient CO2-Reducing Activity of NAD-Dependent Formate Dehydrogenase from Thiobacillus sp KNK65MA for Formate Production from CO2 Gas

NAD-dependent formate dehydrogenase (FDH) from Candida boidinii (CbFDH) has been widely used in various CO2 reduction systems but its practical applications are often impeded due to low CO2-reducing activity. In this study, we demonstrated superior CO2-reducing properties of FDH from Thiobacillus sp. KNK65MA (TsFDH) for production of formate from CO2 gas. To discover more efficient CO2-reducing FDHs than a reference enzyme e. CbFDH, five FDHs were selected with biochemical properties and then, their CO2-reducing activities were evaluated. All FDHs including CbFDH showed better CO2-reducing activities at acidic pHs than at neutral pHs and four FDHs were more active than CbFDH in the CO2 reduction reaction. In particular, the FDH from Thiobacillus sp. KNK65IVIA (TsFDH) exhibited the highest CO2-reducing activity and had a dramatic preference for the reduction reaction, i.e., a 84.2-fold higher ratio of CO2 reduction to formate oxidation in catalytic efficiency (k(cat)/K-B) compared to CbFDH. Formate was produced from CO2 gas using TsFDH and CbFDH, and TsFDH showed a 5.8-fold higher formate production rate than CbFDH. A sequence and structural comparison showed that FDHs with relatively high CO2-reducing activities had elongated N- and C-terminal loops. The experimental results demonstrate that TsFDH can be an alternative to CbFDH as a biocatalyst in CO2 reduction systemsope

Public Sector Budgets: Types, Processes, Implementation and Controls; Issues and Prospects

&lt;p&gt;The budget is a veritable tool for effective service delivery to the people by any government, the performance of budget with the Nigerian ministries has been an issue of concern in recent times. There has been lopsided budget performance with current expenditure performing as high as 100% in a budget year while capital expenditure performing less than expected. With that situation, public sector budget implementation and performance needs a review. This paper critically examines the public sector budget in Nigeria, looking at the various types, processes involved in preparation as well as the implementation cycle and how these have fared against international best practices and standards. The paper adopts Meta-analysis methodology. The study reviewed and anchored on System Analysis Theory. Past empirical literature reviewed and trend analysis showed a huge funding gap that exist among the ministries of government in terms of budget targets and actual budgetary performance over the years and especially in the last five years. The paper via its literature review highlighted some of the issues with Public sector budget in Nigeria viz a viz other nations to include i) Inadequate implementation; ii) Lopsidedness of budgetary allocation; iii) Corruption and wastages; iv) Inability to engage in international best practices; v) Weak or nonexistence of the public participation in the budgeting process; vi) 24 % Open budget index (against 45% global average) and 22% to 26% participation rate within the last seven (7) years among others. The prospects however include its ability to Preparation of a Medium-Term Revenue Framework (MTRF) pursuant to which projected revenue from various oil and non-oil sources is determined over the medium-term; has attained improvement in the rating of international bodies in terms of budget transparency and participation in the past among others. The study concludes with suggestions on ways to better the public sector budgeting process to include: Engage in participatory budgeting (Citizenship Budgeting Process). Engage in Eco and Green Budgeting; Regular Review of budgetary control practices; More deployment of ICT infrastructure in Budgeting processes, monitoring and controls; and Reorientation of stakeholders in the budgeting system (Executive and Legislature)&nbsp;&lt;br&gt;&nbsp;&lt;/p&gt

Intraocular inflammation as the main manifestation of Rickettsia conorii infection

Archimedes LD Agahan1,3, Jenice Torres1, Graciana Fuentes-P&amp;aacute;ez1, Hernan Mart&amp;iacute;nez-Osorio1, Antonio Ordu&amp;ntilde;a2, Margarita Calonge11Ocular Immunology and Uveitis Unit, Institute of Applied Ophthalmobiology (IOBA), 2Microbiology Department, University of Valladolid, Valladolid, Spain; 3Department of Ophthalmology and Visual Sciences, University of the Philippines, Philippine General Hospital, Manila, PhilippinesObjective: To report the clinical features and management of seven cases of intraocular inflammation caused by Rickettsia infection and review published literature.Methods: Rickettsia conorii or Rickettsia spp. infection was diagnosed based on the following criteria: (1) positive serology according to the European Guidelines, (2) titer normalization after specific treatment, and (3) complete resolution of ophthalmic disease and accompanying symptoms after antibiotic therapy.Results: Seven patients were referred for uveitis of unknown etiology. All came from regions where Mediterranean spotted fever is prevalent. One patient met the European guidelines criteria for Rickettsia spp. infection, while the other six cases met the criteria for R. conorii infection. The main symptoms were visual loss, floaters, eye redness, photophobia, and ocular pain. Predominant ophthalmic signs included vasculitis, choroiditis, vitritis, and macular edema. All patients required antibiotic treatment that resulted in the remission of the infection. Doxycycline was the first choice and the only antibiotic used to treat four patients. One patient needed ciprofloxacin as a second antibiotic after not responding to doxycycline. Two patients had doxycycline as a second antibiotic after not responding primarily to sulfonamides (which had been given after 2&amp;ndash;3 days of doxycycline gastric intolerance); one of these patients needed ciprofloxacin as a third antibiotic.Conclusion: Intraocular inflammation can occur as the main manifestation of Rickettsia conorii or Rickettsia spp. infection. It should be considered as a differential diagnosis for uveitis especially for patients living in countries where this infection is endemic in the world. Antibiotic treatment remains effective in the management of Rickettsia infection.Keywords: intraocular inflammation, Mediterranean spotted fever, Rickettsia conorii, uveiti

Laboratory spectroscopy of 1, 2-propanediol at millimeter and submillimeter wavelengths

Context. Ethanediol is one of the largest complex organic molecules detected in space thus far. It has been found in different types of molecular clouds. The two propanediol isomers are the next larger diols. Hence, they are viable candidates to be searched for in space. Aims. We wish to provide sufficiently large and accurate sets of spectroscopic parameters of 1, 2-propanediol to facilitate searches for this molecule at millimeter and longer submillimeter wavelengths. Methods. We recorded rotational spectra of 1, 2-propanediol in three wide frequency windows between 38 and 400 GHz. Results. We made extensive assignments for the three lowest energy conformers to yield spectroscopic parameters up to eighth order of angular momentum. Conclusions. Our present data will be helpful for identifying 1, 2-propanediol at moderate submillimeter or longer wavelengths with radio telescope arrays such as ALMA, NOEMA, or EVLA. In particular, its detection with ALMA in sources, in which ethanediol was detected, appears to be promising

Millimetre-wave spectrum of the singly deuterated isotopologues of anti-ethanol

The rotational spectra of three mono-substituted deuterium sisotopologues of the anti conformer of ethanol have been measured between 35 and 500 GHz. One of these, CH2DCH2OH, exists as two distinct conformers according to the position of the deuterium atom with respect to the molecular skeleton. The dataset was constrained for fitting with a standard Watson-S reduction Hamiltonian by rejecting transitions from high-lying states, which appear to be perturbed by the gauche states, and by averaging some small methyl torsional splits. This treatment is compatible with the one we employed for the C-13 containing isotopologues of ethanol, and with the needs for a first search for these species in the interstellar medium, in particular in spectra taken by ALMA. For this purpose an appropriate set of predictions is given. (C) 2015 Elsevier Inc. All rights reserved