Co-solute assistance in refolding of recombinant proteins

Prokaryotic expression system is the most widely used host for the production of recombinant proteins but inclusion body formation is a major bottleneck in the production of recombinant proteins in prokaryotic cells, especially in Escherichia coli. In vitro refolding of inclusion body into the the proteins with native conformations is a solution for this problem but there is a need for optimization of condition for each protein specifically. Several approaches have been described for in vitro refolding; most of them involve the use of additives for assisting correct refolding. Co-solutes play a major role in refolding process and can be classified according to their function as, aggregation suppressors and folding enhancers. This study presents a review of additives that are used in refolding process of insoluble recombinant proteins in small scale and industrial process.Key words: Refolding, protein aggregation, low-molecular-weight additives, arginine

Criticality and oscillatory behavior in non-Markovian Contact Process

A Non-Markovian generalization of one-dimensional Contact Process (CP) is being introduced in which every particle has an age and will be annihilated at its maximum age $\tau$. There is an absorbing state phase transition which is controlled by this parameter. The model can demonstrate oscillatory behavior in its approach to the stationary state. These oscillations are also present in the mean-field approximation which is a first-order differential equation with time-delay. Studying dynamical critical exponents suggests that the model belongs to the DP universlity class.Comment: 4 pages, 5 figures, to be published in Phys. Rev.

Surface Analysis of Coal Indicating Neutral Red Enhances the Precursor Steps of Methanogenesis

Artificially stimulated, high-yield microbial production of methane from coal is a challenging problem that continues to generate research interest. Decomposition of organic matter and production of methane from coal are the results of multiple redox reactions carried out by different communities of bacteria and archaea. Recent work by our group (Beckmann et al., 2015) demonstrated that the presence of the redox-mediating molecule neutral red, in its crystalline form on a coal surface, can increase methane production. However, hydrolysis and the acetogenesis of the coal surface are essential precursor steps for methane production by archaea. Acetogenesis is the preparation phase of methanogenesis because methanogens can only assimilate acetate, CO2 and H2 among the products formed during this process. In the present study, the surface chemical analysis of neutral red treated coal using attenuated total reflectance-fourier transform infrared (ATR-FTIR) spectroscopy and X-ray photoelectron spectroscopy (XPS) demonstrate that the acetate production and resulting oxidation of the coal only occurred at few nanometers into the coal surface (at the nanoscale <5 nm). We observed that in the presence of neutral red and groundwater microbes, acetate signals in coal surface chemistry increased. This is the first evidence suggesting that neutral red enhances the biological conversion of coal into acetate. Microscopy demonstrated that neutral red crystals were co-localize with cells at the surface of coal in groundwater. This is consistent with neutral red crystals serving as a redox hub, concentrating and distributing reducing equivalents amongst the microbial community. In this study, the chemical changes of neutral red treated coal indicated that neutral red doubles the concentration of acetate over the control (coal without neutral red), emphasizing the importance of maximizing the fracture surface coverage of this redox mediator. Overall, results suggested that, neutral red not only can benefit acetoclastic methanogens, but also the fermentative and acetogenic bacteria involved in generating acetate

First Detection of Nosema ceranae, a Microsporidian Protozoa of European Honeybees (Apis mellifera) In Iran

Background: Nosemosis of European honey bee (Apis mellifera) is present in bee colonies world­wide. Until recently, Nosema apis had been regarded as the causative agent of the disease, that causes heavy economic losses in apicultures. Nosema ceranae is an emerging microsporidian para­site of European honeybees, A. mellifera, but its distribution is not well known. Previously, nosemosis in honeybees in Iran was attributed exclusively to N. apis.Methods: Six Nosema positive samples (determined from light microscopy of spores) of adult worker bees from one province of Iran (Savadkouh- Mazandaran, northern Iran) were tested to determine Nosema species using previously- developed PCR primers of the 16 S rRNA gene. As it is difficult to distinguish N. ceranae and N. apis morphologically, a PCR assay based on 16 S ribosomal RNA has been used to differentiate N. apis and N. ceranae.Results: Only N. ceranae was found in all samples, indicating that this species present in Iran apiar­ies.Conclusion: This is the first report of N. ceranae in colonies of A. mellifera in Iran. It seems that intensive surveys are needed to determine the distribution and prevalence of N. ceranae in differ­ent regions of Iran

Age, growth and spawning season of Luciobarbus esocinus Heckel, 1843 in Gamasiab River, Iran

In the current paper reproductive strategy and growth rates of Luciobarbus esocinus were investigated from 2010 to 2012, in Gamasiab River, Kermanshah, Iran. Samples were collected seasonally by gillnet and electro-fisher device. Sex ratio was evaluated from 162 individuals and the percentage of male and female individuals were 54.76% and 45.24% respectively. Age structure was distributed between I and V and most frequency numbers for males and females belonged to II and IV age group, respectively. Length-weight relationship was computed as W=8E-05TL^2.6546 for all individuals. Mean total length and weight was 333.82±13.10 mm and 297.67±4.85 g for males, and 371.5±21.17 mm and 307.85±7.64 g for females. The von-Bertalanffy growth models of L. esocinus were described as Lt =357.14×[1-e^(-0.77×(t-0.090))] for males, Lt =367.02×[1-e^(-0.78×(t–0.094))] for females and Lt =363.57×[1-e^(-0.76×(t –0.087))] for both sexes. Results indicated that growth factors were highly similar in spring, summer and winter. CDA analysis revealed that the total and gonad weight are the main factors in discriminant analysis. The highest GSI values were found in spring for females and winter for males, however, it decreased rapidly after spring in females. Maximum growth rates were observed in early ages (I-II) and depleted with ages. K value and L∞ were determined as 0.77±0.09 Y^-1 and 357.14±10.38 mm for males and 0.78±0.24 Y^-1 and 367.02±24.15 mm for females, respectively. In conclusion this study provided basic information for stock assessment and management of L. esocinus in Gamasiab Reservoir

A Simple Approach to Dynamic Material Balance in Gas-Condensate Reservoirs

In traditional material balance calculations, shut-in well pressure data are used to determine average reservoir pressure while recent techniques do not require the well to be shut-in and use instead flowing well pressure-rate data. These methods, which are known as “dynamic” material balance, are developed for single-phase flow (oil or gas) in reservoirs. However, utilization of such methods for gas-condensate reservoirs may create significant errors in prediction of average reservoir pressure due to violation of the single-phase assumption in such reservoirs. In a previous work, a method for production data analysis in gas-condensate reservoirs was developed. The method required standard gas production rate, producing gas-oil ratio, flowing well pressure, CVD data and relative permeability curves. This paper presents a new technique which does not need relative permeability curves and flowing well pressure. In this method, the producing oil-gas ratio is interpolated in the vaporized oil in gas phase (Rv) versus pressure (p) data in the CVD table and the corresponding pressure is located. The parameter pressure/two-phase deviation factor (p/ztp) is then evaluated at the determined pressure points and is plotted versus produced moles (np) which forms a straight line. The nature of this plot is such that its extrapolation to point where p/ztp = 0 will give initial moles in place. Putting initial pressure/initial two-phase deviation factor (pi/ztp,i) (known parameter) and estimated initial moles (ni) into the material balance equation, average reservoir pressure can be determined. A main assumption behind the method is that the region where both gas and condensate phases are mobile is of negligible size compared to the reservoir. The approach is quite simple and calculations are much easier than the previous work. It provides a practical engineering tool for industry studies as it requires data which are generally available in normal production operations. However, it is only applicable when average reservoir pressure approaches dew point pressure and falls below it. The methodology is validated using synthetic production data for several examples. In addition, the method is evaluated through estimation of average reservoir pressure and original gas in place from actual field data. The results show a fairly good agreement in gas in place obtained from the new method and that of volumetrically calculated value for this field

A double masked randomised 4-week, placebo-controlled study in the USA, Thailand and Taiwan to compare the efficacy of oral valganciclovir and topical 2% ganciclovir in the treatment of cytomegalovirus anterior uveitis: study protocol.

IntroductionCytomegalovirus (CMV) anterior uveitis is a recognised cause of anterior uveitis in immunocompetent patients and is preventable cause of vision loss. Ocular sequelae include corneal endothelial damage which can cause corneal oedema and failure, as well as glaucoma. Recurrences of inflammation are common and therefore patients are often exposed to long-term therapy. Oral therapy is available in the form of valganciclovir, although with the caveat of systemic side effects such as bone marrow suppression and renal failure necessitating regular interval laboratory monitoring. Recent reports have demonstrated that topical 2% ganciclovir solution may offer promising treatment outcomes in patients with CMV anterior uveitis with superior safety, cost-effectiveness and convenience profiles. An investigation into the relative equipoise of these therapies is warranted for these reasons.Methods and analysisThe Systemic and Topical Control of Cytomegalovirus Anterior uveitis: Treatment Outcomes (STACCATO) trial is designed as a multicentre, block randomised by site, double-masked, placebo-controlled trial comparing the efficacy of oral valganciclovir, 2% topical ganciclovir and placebo in treating PCR-proven CMV anterior uveitis. Participant clinical evaluation will occur at three study time points by a masked study ophthalmologist over a 28-day period to assess resolution of ocular inflammation (secondary outcome). A control group will provide additional information about the possible impact that the infected host's immune response may play in controlling local viral replication. The primary analysis is an analysis of covariance (three arms) correcting for baseline to compare quantitative CMV viral load in the anterior chamber (AC) aqueous fluid before and 7 days after treatment.Ethics and disseminationThe University of California San Francisco Committee on Human Research and the Khon Kaen University Institutional Review Board have given ethical approval. The results of this trial will be presented at local and international meetings and submitted for peer-reviewed journals for publication.Trial registration numberNCT03576898

Impact of glycine betaine on salinity tolerance of stevia (Stevia rebaudiana Bertoni) under in vitro condition

Stevia (Stevia rebaudiana Bertoni), with great potential as a natural sweeteners source, has a high content of sweeteners, which are up to 150 times sweeter than sugar, but virtually with no calories. Stevia also suitable to be cultivated in semiarid climates and coastal areas, which are characterized by the low quality of the irrigation water. Soil salinity occupies a prominent place among the soil problems that threaten the sustainability of agriculture over a vast area in the world. Glycine betaine is an osmoprotectant, that plays an important role and accumulates rapidly in many plants during salinity or drought stress. In order to evaluation of glycine betaine amending effects on salinity stress in stevia under in vitro condition, a factorial experiment was conducted in 2015. Four NaCl levels, including 0, 50, 75 and 100 mM, along with 0, 1, 12.5, 25 and 50 mM of glycine betaine concentrations were used in Murashige and Skoog (MS) medium. The results showed that salinity levels had significant reduction effects on plant height, root length, shoot fresh weight, number of leaf, total chlorophyll, rebaudioside A and stevioside of the stevia genotype. Due to increasing of glycine betaine, levels all the traits were increased. Owing to amending effect of glycine betaine, its high concentrations made less hazarding effects of salinity on the researched traits. The highest mean value of rebaudioside A (10.62rt) and stevioside (23.38rt) determined at 50 mM of glycine betaine with 0 mM of NaCl concentration

Diagnostic Armamentarium of Infectious Keratitis: A Comprehensive Review

Infectious keratitis (IK) represents the leading cause of corneal blindness worldwide, particularly in developing countries. A good outcome of IK is contingent upon timely and accurate diagnosis followed by appropriate interventions. Currently, IK is primarily diagnosed on clinical grounds supplemented by microbiological investigations such as microscopic examination with stains, and culture and sensitivity testing. Although this is the most widely accepted practice adopted in most regions, such an approach is challenged by several factors, including indistinguishable clinical features shared among different causative organisms, polymicrobial infection, long diagnostic turnaround time, and variably low culture positivity rate. In this review, we aim to provide a comprehensive overview of the current diagnostic armamentarium of IK, encompassing conventional microbiological investigations, molecular diagnostics (including polymerase chain reaction and mass spectrometry), and imaging modalities (including anterior segment optical coherence tomography and in vivo confocal microscopy). We also highlight the potential roles of emerging technologies such as next-generation sequencing, artificial intelligence-assisted platforms. and tele-medicine in shaping the future diagnostic landscape of IK

Optical Sum Rule in Finite Bands

In a single finite electronic band the total optical spectral weight or optical sum carries information on the interactions involved between the charge carriers as well as on their band structure. It varies with temperature as well as with impurity scattering. The single band optical sum also bears some relationship to the charge carrier kinetic energy and, thus, can potentially provide useful information, particularly on its change as the charge carriers go from normal to superconducting state. Here we review the considerable advances that have recently been made in the context of high $T_c$ oxides, both theoretical and experimental.Comment: Review article accepted for publication in J. Low Temp. Phys. 29 pages, 33 figure