bluuebunny/crossref_metadata_2025 huggingface dataset:
Engineering NAD+ availability for Escherichia coli whole-cell biocatalysis: a case study for dihydroxyacetone production
Authors: Zhou Yongjin J , Yang Wei , Wang Lei , Zhu Zhiwei , Zhang Sufang , Zhao Zongbao K
Published: November 2013

Background

Whole-cell redox biocatalysis has been intensively explored for the production of valuable compounds because excellent selectivity is routinely achieved. Although the cellular cofactor level, redox state and the corresponding enzymatic activity are expected to have major effects on the performance of the biocatalysts, our ability remains limited to predict the outcome upon variation of those factors as well as the relationship among them.

Results

In order to investigate the effects of cofactor availability on whole-cell redox biocatalysis, we devised recombinant Escherichia coli strains for the production of dihydroxyacetone (DHA) catalyzed by the NAD+-dependent glycerol dehydrogenase (GldA). In this model system, a water-forming NAD+ oxidase (NOX) and a NAD+ transporter (NTT4) were also co-expressed for cofactor regeneration and extracellular NAD+ uptake, respectively. We found that cellular cofactor level, NAD+/NADH ratio and NOX activity were not only strain-dependent, but also growth condition-dependent, leading to significant differences in specific DHA titer among different whole-cell biocatalysts. The host E. coli DH5α had the highest DHA specific titer of 0.81 g/gDCW with the highest NAD+/NADH ratio of 6.7 and NOX activity of 3900 U. The biocatalyst had a higher activity when induced with IPTG at 37°C for 8 h compared with those at 30°C for 8 h and 18 h. When cells were transformed with the ntt4 gene, feeding NAD+ during the cell culture stage increased cellular NAD(H) level by 1.44 fold and DHA specific titer by 1.58 fold to 2.13 g/gDCW. Supplementing NAD+ during the biotransformation stage was also beneficial to cellular NAD(H) level and DHA production, and the highest DHA productivity reached 0.76 g/gDCW/h. Cellular NAD(H) level, NAD+/NADH ratio, and NOX and GldA activity dropped over time during the biotransformation process.

Conclusions

High NAD+/NADH ratio driving by NOX was very important for DHA production. Once cofactor was efficiently cycled, high cellular NAD(H) level was also beneficial for whole-cell redox biocatalysis. Our results indicated that NAD+ transporter could be applied to manipulate redox cofactor level for biocatalysis. Moreover, we suggested that genetically designed redox transformation should be carefully profiled for further optimizing whole-cell biocatalysis.
Downregulation of the H-2Kd gene by siRNA affects the cytotoxicity of murine LAK cells
Authors: Liu Xin , Cui Xin , Shan Ningning , Li Ying , Fang Xiaosheng , Ding Mei , Wang Xin
Published: November 2013

To investigate the effect of the H-2Kdgene on the lymphocyte membrane, we constructed a small interfering RNA (siRNA) that targets the H-2Kdgene and compared the cytotoxicity of mouse lymphokine-activated killer (LAK) cells with different H-2Kdexpression states. H-2Kd-targeting siRNA was transfected into spleen lymphocytes of BALB/C mice. Flow cytometry (FCM) was then performed to examine the expression of the H-2Kdgene in the transfected and control cells. Additionally, the cytotoxicity of the transfected cells toward the H22 and K562 cell lines was evaluated in vitro using the LDH release assay. H-2Kd-targeting siRNA significantly reduced the expression levels of the target protein, whereas pure transMessenger and non-silencing siRNA did not inhibit H-2Kdexpression at the concentrations tested. The cytotoxicity of siRNA-treated LAK cells toward H22 and K562 cells was reduced significantly. The knockdown of H-2Kdgene expression by siRNA may be associated with LAK cell cytotoxicity toward neoplasm cell lines.
Evaluation of parasite subpopulations and genetic diversity of the msp1, msp2 and glurp genes during and following artesunate monotherapy treatment of Plasmodium falciparum malaria in Western Cambodia
Authors: Gosi Panita , Lanteri Charlotte A , Tyner Stuart D , Se Youry , Lon Chanthap , Spring Michele , Char Mengchuor , Sea Darapiseth , Sriwichai Sabaithip , Surasri Sittidech , Wongarunkochakorn Saowaluk , Pidtana Kingkan , Walsh Douglas S , Fukuda Mark M , Manning Jessica , Saunders David L , Bethell Delia
Published: November 2013

Background

Despite widespread coverage of the emergence of artemisinin resistance, relatively little is known about the parasite populations responsible. The use of PCR genotyping around the highly polymorphic Plasmodium falciparum msp1, msp2 and glurp genes has become well established both to describe variability in alleles within a population of parasites, as well as classify treatment outcome in cases of recurrent disease. The primary objective was to assess the emergence of minority parasite clones during seven days of artesunate (AS) treatment in a location with established artemisinin resistance. An additional objective was to investigate whether the classification of clinical outcomes remained valid when additional genotyping was performed.

Methods

Blood for parasite genotyping was collected from 143 adult patients presenting with uncomplicated falciparum malaria during a clinical trial of AS monotherapy in Western Cambodia. Nested allelic type-specific amplification of the genes encoding the merozoite surface proteins 1 and 2 (msp1 and msp2) and the glutamate-rich protein (glurp) was performed at baseline, daily during seven days of treatment, and again at failure. Allelic variants were analysed with respect to the size of polymorphisms using Quantity One software to enable identification of polyclonal infections.

Results

Considerable variation of msp2 alleles but well-conserved msp1 and glurp were identified. At baseline, 31% of infections were polyclonal for one or more genes. Patients with recurrent malaria were significantly more likely to have polyclonal infections than patients without recurrence (seven of nine versus 36 of 127, p = 0.004). Emergence of minority alleles during treatment was detected in only one of twenty-three cases defined as being artemisinin resistant. Moreover, daily genotyping did not alter the final outcome classification in any recurrent cases.

Conclusions

The parasites responsible for artemisinin-resistant malaria in a clinical trial in Western Cambodia comprise the dominant clones of acute malaria infections rather than minority clones emerging during treatment. Additional genotyping during therapy was not beneficial. Disproportionately high rates of polyclonal infections in cases of recurrence suggest complex infections lead to poor treatment outcomes. Current research objectives should be broadened to include identification and follow-up of recurrent polyclonal infections so as to define their role as potential agents of emerging resistance.
Acetylcholinesterase (Ace-1) target site mutation 119S is strongly diagnostic of carbamate and organophosphate resistance in Anopheles gambiae s.s. and Anopheles coluzzii across southern Ghana
Authors: Essandoh John , Yawson Alexander E , Weetman David
Published: November 2013

Background

With high DDT resistance present throughout much of West Africa, carbamates and organophosphates are increasingly important alternatives to pyrethroids for indoor residual spraying (IRS). Though less widespread, resistance to both of these alternative insecticide classes has also been documented within the Anopheles gambiae species pair (formerly the M and S molecular forms) in West Africa. To manage insecticide efficacy, it is important to predict how and where resistance is likely to occur and spread, which could be aided by using molecular diagnostics with high predictive value.

Methods

Anopheles coluzzii and An. gambiae s.s. were collected from 18 sites throughout southern Ghana and bioassayed with bendiocarb, the most commonly applied carbamate, and an organophosphate, fenitrothion. The Ace-1 target site substitution G119S was genotyped by qPCR.

Results

Fenitrothion induced higher mortality than bendiocarb, though phenotypes correlated strongly across populations. Ace-1 119S was found at much higher frequency in An. gambiae s.s than An. coluzzii, exceeding 90% in a population from Greater Accra, the highest frequency reported to date. Ace-1 G119S was very strongly associated with resistance to both insecticides, providing high predictive power for diagnosis, though with some evidence for a differential effect between molecular forms for bendiocarb. Sequencing of the gene revealed a lack of variation in resistant alleles precluding determination of origin, but Ace-1 copy number variation was detected for the first time in Ghana.

Conclusions

The results validate G119S as a useful diagnostic of organophosphate and carbamate resistance within and among populations, whilst highlighting the potential for an aggregate nature of Ace-1 genotypes, which may comprise both single-copy and duplicated genes. Further work is now required to determine the distribution and resistance-association of Ace-1 duplication.
Dynamics of vitamin D in patients with mild or inactive inflammatory bowel disease and their families
Authors: Grunbaum Avigyle , Holcroft Christina , Heilpern Debra , Gladman Stephanie , Burstein Barry , Menard Maryse , Al-Abbad Jasim , Cassoff Jamie , MacNamara Elizabeth , Gordon Philip H , Szilagyi Andrew
Published: November 2013

Background

25(OH) vitamin D levels may be low in patients with moderately or severely active inflammatory bowel diseases (IBD: Crohn’s disease and Idiopathic Ulcerative Colitis) but this is less clear in patients with mild or inactive IBD. Furthermore there is limited information of any family influence on 25(OH) vitamin D levels in IBD. As a possible risk factor we hypothesize that vitamin D levels may also be low in families of IBD patients.

Objectives

To evaluate 25[OH] vitamin D levels in patients with IBD in remission or with mild activity. A second objective is to evaluate whether there are relationships within IBD family units of 25[OH] vitamin D and what are the influences associated with these levels.

Methods

Participants underwent medical history, physical examination and a 114 item diet questionnaire. Serum 25[OH] vitamin D was measured, using a radioimmunoassay kit, (replete ≥ 75, insufficient 50–74, deficient < 25–50, or severely deficient < 25 nmol/L). Associations between 25[OH] vitamin D and twenty variables were evaluated using univariate regression. Multivariable analysis was also applied and intrafamilial dynamics were assessed.

Results

55 patients and 48 controls with their respective families participated (N206). 25[OH] vitamin D levels between patients and controls were similar (71.2 ± 32.8 vs. 68.3 ±26.2 nmol/L). Vitamin D supplements significantly increased intake but correlation with serum 25[OH] vitamin D was significant only during non sunny months among patients. Within family units, patients’ families had mean replete levels (82.3 ± 34.2 nmol/L) and a modest correlation emerged during sunny months between patients and family (r2 =0.209 p = 0.032). These relationships were less robust and non significant in controls and their families.

Conclusions

In patients with mild or inactive IBD 25[OH] vitamin D levels are less than ideal but are similar to controls. Taken together collectively, the results of this study suggest that patient family dynamics may be different in IBD units from that in control family units. However contrary to the hypothesis, intra familial vitamin D dynamics do not pose additional risks for development of IBD.
First-line treatment patterns and lipid target levels attainment in very high cardiovascular risk outpatients
Authors: Xanthopoulou Ioanna , Davlouros Periklis , Siahos Simos , Perperis Angelos , Zaharioglou Evangelia , Alexopoulos Dimitrios
Published: November 2013

Objectives

Previous studies have demonstrated gaps in achievement of low-density lipoprotein-cholesterol (LDL-C) goals among patients at very high cardiovascular risk. We aimed to investigate lipid treatment patterns, rates and predictors of lipid targets attainment, in such outpatients in an urban area of Greece.

Methods

This was a prospective observational study, conducted in 19 outpatient clinics of Western Greece. We recruited patients with established cardiovascular disease (CVD) and/or diabetes mellitus (DM), previously (at least 3 months before baseline assessment) untreated with any lipid lowering medication. Lipid profile assessment was performed at baseline (prior to lipid-lowering treatment initiation) and at follow-up. Lipid lowering treatment choice was at physicians’ discretion and was kept constant until follow-up.

Results

We recruited 712 patients with a mean age 61.4 ± 10.4 years, 68.0% males, 43.0% with DM, 64.7% with prior coronary artery disease-CAD. In total, 237/712 (33.3%) of prescribed regimens were of high or very high LDL-C lowering efficacy and out of them 113/237 (47.7%) comprised a combination of statin and ezetimibe. At follow-up the primary target of LDL-C < 70 mg/dL (1.8 mmol/L) was achieved in 71(10.0%) patients. The secondary target of non-HDL-C < 100 mg/dL (2.6 mmol/L) in the subgroup of patients with DM or increased triglycerides levels (>150 mg/dl or 1.7 mmol/L) was achieved in 45(11.6%) of patients. In multivariate logistic regression analysis (AUC = 0.71, 95% CIs 0.65-0.77, p < 0.001) male gender, smoking, baseline LDL-C and very high potency LDL-C lowering regimen emerged as independent predictors of LDL-C goal attainment (OR = 1.88, 95% CIs 1.03-3.44, p = 0.04, OR = 0.57, 95% CIs 0.33-0.96, p = 0.04, OR = 0.98, 95% CIs 0.98-0.99, p < 0.001 and OR = 2.21, 95% CIs 1.15-4.24, p = 0.02 respectively).

Conclusions

First-line management of dyslipidemia among very-high cardiovascular risk outpatients in Western Greece is unsatisfactory, with the majority of treated individuals failing to attain the LDL-C and non-HDL-C targets. This finding points out the need for intensification of statin treatment in such patients.
Interface coupling-induced enhancement of magnetoimpedance effect in heterogeneous nanobrush by adjusting textures of Co nanowires
Authors: Zhang Yi , Dong Juan , Sun Xiaojun , Liu Qingfang , Wang Jianbo
Published: November 2013

Interface coupling-induced and interface coupling-enhanced magnetoimpedance (MI) effect in heterogeneous nanobrush has been investigated. The nanobrush is composed of Fe25Ni75 nanofilm and textured hexagonal close-packed cobalt nanowire array, respectively fabricated by RF magnetron sputtering and electrochemical deposition. The design of this structure is based on the vortex distribution of magnetic moments in thin film, which can be induced by the exchange coupling effect at the interfaces of the nanobrush. The texture of nanowires plays an important role in the MI effect of the nanobrush, which is regulated by controlling the pH values and temperatures of the deposition process. The ‘parallel’ and ‘perpendicular’ coupling models were used to explain the different MI results of the nanobrush with cobalt nanowires, which have (100) and (002) textures, respectively. The optimized MI effect of the nanobrush brought by (100) nanowires can be magnified by 300% with more than 80%/Oe magnetic sensitivity at a low frequency, which has great application potentials in low-frequency MI sensors.
A new general iterative algorithm with Meir-Keeler contractions for variational inequality problems in q-uniformly smooth Banach spaces
Authors: Guan Jinlin , Hu Changsong , Wang Xin
Published: November 2013

In this paper, we generalize the iterative scheme and extend the space studied in (Fixed Point Theory and Applications 2012:46, 2012). Further, we prove some strong convergence theorems of the new iterative scheme for variational inequality problems in q-uniformly smooth Banach spaces under very mild conditions. Our results improve and extend corresponding ones announced by many others.

MSC:47H09, 47H10.
Transcriptional profiling of feline infectious peritonitis virus infection in CRFK cells and in PBMCs from FIP diagnosed cats
Authors: Harun Mohammad Syamsul Reza , Kuan Choong Oi , Selvarajah Gayathri Thevi , Wei Tan Sheau , Arshad Siti Suri , Bejo Mohd Hair , Omar Abdul Rahman
Published: November 2013

Background

Feline Infectious Peritonitis (FIP) is a lethal systemic disease, caused by the FIP Virus (FIPV); a virulent mutant of Feline Enteric Coronavirus (FECV). Currently, the viruses virulence determinants and host gene expressions during FIPV infection are not fully understood.

Methods

RNA sequencing of Crandell Rees Feline Kidney (CRFK) cells, infected with FIPV strain 79–1146 at 3 hours post infection (h.p.i), were sequenced using the Illumina next generation sequencing approach. Bioinformatic’s analysis, based on Felis catus 2X annotated shotgun reference genome, using CLC bio Genome Workbench mapped both control and infected cell reads to 18899 genes out of 19046 annotated genes. Kal’s Z test statistical analysis was used to analyse the differentially expressed genes from the infected CRFK cells. Real time RT-qPCR was developed for further transcriptional profiling of three genes (PD-1, PD-L1 and A3H) in infected CRFK cells and Peripheral Blood Mononuclear Cells (PBMCs) from healthy and FIP-diseased cats.

Results

Based on Kal’s Z-test, with False Discovery Rate (FDR) <0.05 and >1.99 fold change on gene expressions, a total of 61 genes were differentially expressed by both samples, where 44 genes were up-regulated and the remainder were down-regulated. Most genes were closely clustered together, suggesting a homogeneous expression. The majority of the genes that were significantly regulated, were those associated with monocytes-macrophage and Th1 cell functions, and the regulation of apoptosis. Real time RT-qPCR developed focusing on 2 up-regulated genes (PD-L1 and A3H) together with an apoptosis associated gene PD-1 expressions in FIPV infected CRFK cells and in PBMCs from healthy and FIP diagnosed cats produced concordant results with transcriptome data.

Conclusion

The possible roles of these genes, and their importance in feline coronaviruses infection, are discussed.
Molecular epidemiology of Newcastle disease virus isolates from vaccinated commercial poultry farms in non-epidemic areas of Japan
Authors: Umali Dennis Villaseñor , Ito Hiroshi , Suzuki Terumasa , Shirota Kazutoshi , Katoh Hiromitsu , Ito Toshihiro
Published: November 2013

Background

Newcastle Disease (ND) is a highly contagious and economically devastating disease of poultry. At present, limited molecular epidemiological data are available regarding the causes of ND outbreaks in vaccinated commercial poultry farms. Knowing the genomic characteristics of Newcastle disease virus (NDV) infecting commercial poultry operations in spite of vaccination might give important insights on the infection dynamics of these viruses. In addition, molecular analyses at the subgenotype level and studies on the relationship of Japanese NDVs with other isolates from around the world are lacking. Therefore, in the present study, a molecular epidemiological investigation was conducted to characterize nine NDVs isolated from vaccinated commercial poultry flocks in five different Prefectures in non-epidemic areas of Japan between 1969 and 2002.

Methods

Nucleotide sequencing and phylogenetic studies were performed to characterize the complete fusion (F)-protein gene, 3-prime end of the nucleoprotein (NP)-gene and 5-prime end of the RNA dependent RNA polymerase (L)-gene. Sequence data were compared with 180 NDV strains from GenBank representing different NDV genotypes and subgenotypes from different regions of the world at different time periods. Deduced amino acids were analyzed for homologies, recombination and mutation. Recombination events were estimated using Recombination Detection Program (RDP) version 3.44. Phylogenetic trees were constructed to determine evolutionary relationships among strains.

Results

Mean death time (MDT: 48-56 hr), Intracerebral Pathogenicity Index (ICPI: 1.7-1.9) and deduced amino acid sequences of the F0 proteolytic cleavage site (112RRQKR116) revealed that all nine field isolates were velogenic. Phylogenetic analysis showed that these isolates could be classified into two genetic lineages and three sublineages namely genotypes VIa (lineage 4a), VId (lineage 4d) and VIId (lineage 5d). No recombination events were observed but a point mutation in one of the neutralizing epitope of the F-protein was identified in the field isolates from Japan.

Conclusions

All field isolates from vaccinated commercial poultry in non-epidemic areas of Japan were part of much bigger outbreaks in provinces and regions and, in some cases, continents. In general, four ND panzootics occurred in Japan and that these outbreaks were mostly characterized by co-circulation of genetically distinct virus lineages due to involvements of infected wild birds. The point mutation identified in the field isolates from Japan may be due to escape from vaccine pressure. The identification of such mutation may be useful for future site-directed mutagenesis to understand the dynamics of NDV infection in vaccinated chickens.
Recombinant lentogenic Newcastle disease virus expressing Ebola virus GP infects cells independently of exogenous trypsin and uses macropinocytosis as the major pathway for cell entry
Authors: Wen Zhiyuan , Zhao Bolin , Song Kun , Hu Xule , Chen Weiye , Kong Dongni , Ge Jinying , Bu Zhigao
Published: November 2013

Background

Using reverse genetics, we generated a recombinant low-pathogenic LaSota strain Newcastle disease virus (NDV) expressing the glycoprotein (GP) of Ebola virus (EBOV), designated rLa-EBOVGP, and evaluated its biological characteristic in vivo and in vitro.

Results

The introduction and expression of the EBOV GP gene did not increase the virulence of the NDV vector in poultry or mice. EBOV GP was incorporated into the particle of the vector virus and the recombinant virus rLa-EBOVGP infected cells and spread within them independently of exogenous trypsin. rLa-EBOVGP is more resistant to NDV antiserum than the vector NDV and is moderately sensitive to EBOV GP antiserum. More importantly, infection with rLa-EBOVGP was markedly inhibited by IPA3, indicating that rLa-EBOVGP uses macropinocytosis as the major internalization pathway for cell entry.

Conclusions

The results demonstrate that EBOV GP in recombinant NDV particles functions independently to mediate the viral infection of the host cells and alters the cell-entry pathway.
Hepatic arterial phase and portal venous phase computed tomography for dose calculation of stereotactic body radiation therapy plans in liver cancer: a dosimetric comparison study
Authors: Xiao Jianghong , Li Yan , Jiang Qingfeng , Sun Lan , Henderson Jr Fraser , Wang Yongsheng , Jiang Xiaoqin , Li Guangjun , Chen Nianyong
Published: November 2013

Purpose

To investigate the effect of computed tomography (CT) using hepatic arterial phase (HAP) and portal venous phase (PVP) contrast on dose calculation of stereotactic body radiation therapy (SBRT) for liver cancer.

Methods

Twenty-one patients with liver cancer were studied. HAP, PVP and non-enhanced CTs were performed on subjects scanned in identical positions under active breathing control (ABC). SBRT plans were generated using seven-field three-dimensional conformal radiotherapy (7 F-3D-CRT), seven-field intensity-modulated radiotherapy (7 F-IMRT) and single-arc volumetric modulated arc therapy (VMAT) based on the PVP CT. Plans were copied to the HAP and non-enhanced CTs. Radiation doses calculated from the three phases of CTs were compared with respect to the planning target volume (PTV) and the organs at risk (OAR) using the Friedman test and the Wilcoxon signed ranks test.

Results

SBRT plans calculated from either PVP or HAP CT, including 3D-CRT, IMRT and VMAT plans, demonstrated significantly lower (p <0.05) minimum absorbed doses covering 98%, 95%, 50% and 2% of PTV (D98%, D95%, D50% and D2%) than those calculated from non-enhanced CT. The mean differences between PVP or HAP CT and non-enhanced CT were less than 2% and 1% respectively. All mean dose differences between the three phases of CTs for OARs were less than 2%.

Conclusions

Our data indicate that though the differences in dose calculation between contrast phases are not clinically relevant, dose underestimation (IE, delivery of higher-than-intended doses) resulting from CT using PVP contrast is larger than that resulting from CT using HAP contrast when compared against doses based upon non-contrast CT in SBRT treatment of liver cancer using VMAT, IMRT or 3D-CRT.
Risk factors for endotracheal re-intubation following coronary artery bypass grafting
Authors: Jian Liu , Sheng Shi , Min Yu , Zhongxiang Yuan
Published: November 2013

Background

Endotracheal re-intubation following coronary artery bypass grafting (CABG) is often associated with significant morbidity and mortality. However, few reports have focused on the independent risk factors for re-intubation following CABG. This study aimed to evaluate the independent risk factors for re-intubation following CABG.

Methods

The pre-, intra-, and post-operative materials in patients who had selective and isolated CABG performed on them from January 2004 to July 2012 in our hospital were analyzed retrospectively. Unvariate analysis and logistic regression were used to analyze the risk factor of postoperative re-intubation following CABG.

Results

Among the 1,244 patients investigated, 97 cases suffered from postoperative re-intubation, and the incidence rate of postoperative re-intubation was 7.8%. The in-hospital mortality in the re-intubation group was significantly higher than that in the non-re-intubation group (9.3% versus 1.4%, P = 0.004). Re-intubation also correlated with many negative outcomes such as pneumonia, tracheotomy, acute renal failure, infection of incision, prolonged mechanical ventilation time, prolonged intensive care unit (ICU) stay and prolonged hospital stay. The most commonly cause of re-intubation after CABG was hypoxemia due to cardiogenic and noncardiogenic disease, which accounted for 72.2%. The relative factors of postoperative re-intubation were tested through unvariate analysis and logistic regression, and the associated factors were obtained. The associated factors for re-intubation following CABG included preoperative chronic obstructive pulmonary disease (COPD) (OR = 2.134, 95% CI = 1.472-2.967), preoperative congestive heart failure (CHF) (OR = 2.325, 95% CI = 1.512-3.121), postoperative relative hypoxemia (OR = 2.743, 95% CI = 1.657-3.326), postoperative acute kidney injury (AKI) (OR = 2.976, 95% CI = 2.127-4.023), postoperative total mechanical ventilation time (OR = 1.976, 95% CI = 1.347-2.645).

Conclusion

Preoperative COPD, preoperative CHF, postoperative relative hypoxemia, postoperative AKI and postoperative total mechanical ventilation time were five independent risk factors for re-intubation following CABG.
Tween 40 pretreatment of unwashed water-insoluble solids of reed straw and corn stover pretreated with liquid hot water to obtain high concentrations of bioethanol
Authors: Lu Jie , Li Xuezhi , Yang Ruifeng , Zhao Jian , Qu Yinbo
Published: November 2013

Background

Liquid hot water (LHW) pretreatment is an effective and environmentally friendly method to produce bioethanol with lignocellulosic materials. In our previous study, high ethanol concentration and ethanol yield were obtained from water-insoluble solids (WIS) of reed straw and corn stover pretreated with LHW by using fed-batch semi-simultaneous saccharification and fermentation (S-SSF). However, high cellulase loading and the large amount of wash water possibly limit the practical application of LHW pretreatment. To decrease cellulase loading and the amount of wash water, we performed Tween 40 pretreatment before WIS was subjected to bioethanol fermentation.

Results

Results showed that the optimum conditions of Tween 40 pretreatment were as follows: Tween 40 concentration of 1.5%, WIS-to-Tween 40 ratio of 1:10 (w/v), and pretreatment time of 1 hour at ambient temperature. After Tween 40 pretreatment, cellulase loading could be greatly reduced. After Tween 40 pretreatment, the residual liquid could be recycled for utilization but slightly affected ethanol concentration and yield. The unwashed WIS could obtain a high ethanol concentration of 56.28 g/L (reed straw) and 52.26 g/L (corn stover) by Tween 40 pretreatment using fed-batch S-SSF. Ethanol yield reached a maximum of 69.1% (reed straw) and 71.1% (corn stover).

Conclusions

Tween 40 pretreatment was a very effective and less costly method with unwashed WIS. This pretreatment could greatly reduce cellulase loading and save wash water. Higher ethanol concentration was obtained almost without reducing ethanol yield.
Intestinal parasitosis in school children of Lalitpur district of Nepal
Authors: Tandukar Sarmila , Ansari Shamshul , Adhikari Nabaraj , Shrestha Anisha , Gautam Jyotshana , Sharma Binita , Rajbhandari Deepak , Gautam Shikshya , Nepal Hari Prasad , Sherchand Jeevan B
Published: November 2013

Background

Enteric parasites are the most common cause of parasitic diseases and cause significant morbidity and mortality, particularly in developing countries like Nepal. The objective of this study was to estimate the prevalence and risk factors of intestinal parasitic infections among school going children of Lalitpur district of Nepal.

Methods

A total of 1392 stool samples were collected from school children of two government, two private and two community schools of the same district. The stool samples were examined for evidence of parasitic infections by direct microscopy and confirmed by concentration methods (formal ether sedimentation technique or floatation technique by using Sheather’s sugar solution). Modified Ziehl-Neelsen (ZN) staining was performed for the detection of coccidian parasites.

Results

Prevalence of intestinal parasitosis was found to be 16.7%. The highest prevalence rate was seen with Giardia lamblia (7.4%) followed by Entamoeba histolytica (3.4%) and Cyclospora cayetanensis (1.6%). Children aged 11–15 years and the ones belonging to family of agriculture workers were most commonly affected. Hand washing practice and type of drinking water also showed significant difference.

Conclusions

The burden of parasitic infections among the school children, coupled with the poor sanitary conditions in the schools, should be regarded as an issue of public health priority and demands for effective school health programs involving periodic health education and screening.
Immunoglobulin A nephropathy with massive paramesangial deposits caused by anti-vascular endothelial growth factor therapy for metastatic rectal cancer: a case report and review of the literature
Authors: Yahata Mayumi , Nakaya Izaya , Sakuma Tsutomu , Sato Hiroshi , Aoki Shigehisa , Soma Jun
Published: November 2013

Background

Bevacizumab, a recombinant humanized monoclonal antibody for vascular endothelial growth factor, has been widely used in various cancers offering substantial clinical benefit. It is reportedly associated with development of high-grade proteinuria and nephrotic syndrome with the histology of thrombotic microangiopathy, but there has been no report describing the development of immunoglobulin A nephropathy in bevacizumab-treated patients.

Case presentation

A 68-year-old man with metastatic rectal cancer was treated with bevacizumab. He presented with hematuria and proteinuria 15 and 17 months, respectively, after bevacizumab initiation. Bevacizumab was stopped at 17 months. Renal biopsy at 19 months revealed immunoglobulin A nephropathy, with numerous paramesangial hemispherical deposits and thrombotic microangiopathy. Electron microscopy showed numerous paramesangial electron-dense deposits of various sizes, and subendothelial injuries. Proteinuria almost completely resolved 8 months after bevacizumab cessation, although hematuria persisted. Follow-up renal biopsy 11 months after bevacizumab cessation showed a marked decrease in mesangial immunoglobulin A deposits and paramesangial electron-dense deposits, which correlated with a gradual decrease in serum immunoglobulin A.

Conclusion

This is the first case report that confirmed histologically the development and resolution of immunoglobulin A nephropathy during and after bevacizumab therapy. This case shows that there may be other mechanisms of glomerular injury by bevacizumab besides glomerular endothelial injury leading to thrombotic microangiopathy.
Carboplatin and oxaliplatin in sequenced combination with bortezomib in ovarian tumour models
Authors: Al-Eisawi Zaynab , Beale Philip , Chan Charles , Yu Jun Q , Huq Fazlul
Published: November 2013

Background

Ovarian cancer remains an on-going challenge mainly due to the development of drug resistance and also because the cancer is likely to have metastasized at the time of diagnosis. Currently, chemotherapy based on platinum drugs such as cisplatin is the primary treatment for the disease. Copper transporter 1 is involved in the transport of cisplatin into the cell, but is also down-regulated by the drug. Bortezomib, a proteasome inhibitor, has been reported to block this platinum-induced down-regulation of CTR1, so that in the presence of bortezomib, the cellular uptake of platinum drugs may be increased. Increased platinum accumulation may result in increased platinum − DNA binding so that the platinum drug in combination with bortezomib may produce enhanced cell kill.

Methods

In this study the efficacy of the sequential combinations of carboplatin, oxaliplatin and atrans-platinum compound coded as CH1 with BORT on the human ovarian A2780, A2780cisR, A2780ZD0473Rand SKOV-3 cancer cell lines was evaluated. The levels of cellular platinum accumulation and platinum-DNA binding were determined following the treatment with these combinations. In order to investigate the effect of the combinations of the formation of ROS, the total and oxidized glutathione levels were also determined.

Results

Prevention of copper transporter 1 degradation by bortezomib is found to enhance the cellular accumulation of platinum, the level of Platinum − DNA binding and increases oxidative stress especially in the resistant cell lines.

Conclusions

The results suggest that the prevention of CTR1 degradation by bortezomib may be playing a major role in increasing the cellular uptake of platinum drugs and platinum-DNA binding level. Furthermore, the generation of oxidative stress appears to be a major contributor to the enhanced cell kill.
Nanoscale elucidation of Na,K-ATPase isoforms in dendritic spines
Authors: Liebmann Thomas , Blom Hans , Aperia Anita , Brismar Hjalmar
Published: November 2013

Background

The dimensions of neuronal synapses suggest that optical super-resolution imaging methods are necessary for thorough investigation of protein distributions and interactions. Nanoscopic evaluation of neuronal samples has presented practical hurdles, but advancing methods are making synaptic protein topology and quantification measurements feasible. This work explores the application of Photoactivated Localization Microscopy (PALM) pointillistic super-resolution imaging for investigation of the membrane bound sodium pump, the Na,K-ATPase, in matured neurons.

Results

Two isoforms of the sodium pump (ATP1a1 and ATP1a3) were studied in cultured neurons using the PALM-compatible fluorescent proteins PAGFP and mEos. Nanoscopic imaging reveals a compartmentalized distribution of sodium pumps in dendritic spines. Several nanoclusters of pumps are typically found in the spine head and fewer in the spine neck. The density of sodium pumps was estimated from a quantification of detected single molecules at 450–650 pump copies/μm2 in the spine heads.

Conclusions

We have utilized PALM for dissection of nanoscale localization in mature cultured neurons and demonstrated similar topology and quantification estimates with PAGFP and mEos. PALM topology assessments of the sodium pump appeared similar to previous STED studies, though quantification estimates varied, implying that labeling strategies, sample analysis and choice of nanoscopic imaging method can be critical factors for correct molecular quantification.
Proposed design for the PGAA facility at the TRIGA IPR-R1 research reactor
Authors: Guerra Bruno T , Jacimovic Radojko , Menezes Maria Angela BC , Leal Alexandre S
Published: November 2013

Background

This work presents an initial proposed design of a Prompt Gamma Activation Analysis (PGAA) facility to be installed at the TRIGA IPR-R1, a 60 years old research reactor of the Centre of Development of Nuclear Technology (CDTN) in Brazil. The basic characteristics of the facility and the results of the neutron flux are presented and discussed.

Findings

The proposed design is based on a quasi vertical tube as a neutron guide from the reactor core, inside the reactor pool, 6 m below the room’s level where shall be located the rack containing the set sample/detector/shielding. The evaluation of the thermal and epithermal neutron flux in the sample position was done considering the experimental data obtained from a vertical neutron guide, already existent in the reactor, and the simulated model for the facility.

Methods

The experimental determination of the neutron flux was obtained through the standard procedure of using Au monitors in different positions of the vertical tube. In order to validate both, this experiment and calculations of the simulated model, the flux was also determined in different positions in the core used for sample irradiation. The model of the system was developed using the Monte Carlo code MCNP5.

Conclusion

The preliminary results suggest the possibility of obtaining a beam with minimum thermal flux of magnitude 106 cm-2 s-1, which confirm the technical feasibility of the installation of PGAA at the TRIGA IPR-R1 reactor. This beam would open new possibilities for enhancing the applications using the reactor.
A hybrid fuzzy stochastic analytical hierarchy process (FSAHP) approach for evaluating ballast water treatment technologies
Authors: Jing Liang , Chen Bing , Zhang Baiyu , Peng Hongxuan
Published: November 2013

Background

Environmental decisions can be complex because of the inherent trade-offs among environmental, social, ecological, and economic factors. This paper presents a novel hybrid fuzzy stochastic analytical hierarchy process (FSAHP) approach to aid decision making by incorporating fuzzy and stochastic uncertainty into the traditional analytic hierarchy process (AHP). A case study related to ballast water management is used to demonstrate the applicability of the proposed approach. Nine experts from government ministries and academic institutions are invited to evaluate five treatment technologies (i.e., heat treatment, ultraviolet, ozone, ultrasound, and biocide) based on a number of criteria such as efficacy, capital cost, and human risk.

Results

The experts’ preferences over the set of alternatives are represented as linguistic terms instead of numerical values. The beta-PERT distribution is adopted to approximate the probability density functions of the values of their inputs. Statistical analysis indicates that ultraviolet has the highest score (0.22–0.24) in most replications and its overlap with the second-best alternative is statistically negligible. Ozone, ultrasound, and heat treatment are mostly found as the second-, third-, and fourth-best alternatives with considerable overlaps that may be reduced if more experts are involved.

Conclusions

As compared with the traditional AHP, the proposed FSAHP approach can not only take into account linguistic information but also capture the uncertainty associated with insufficient information and biased opinions in group decision-making problems.
CD14hiHLA-DRdim macrophages, with a resemblance to classical blood monocytes, dominate inflamed mucosa in Crohn's disease
Authors: Thiesen Susanne , Janciauskiene Sabina , Uronen-Hansson Heli , Agace William , Högerkorp Carl-Magnus , Spee Pieter , Håkansson Katarina , Grip Olof
Published: November 2013

Intestinal Mϕ play an important role in maintaining gut homeostasis. However, little is known about these cells, their precursors, and their role in intestinal inflammation. Here, we characterize the CD14+ mononuclear cell populations in intestinal mucosa and blood in patients with CD. Among the LP CD14+ Mϕ, we identified three distinct HLA-DR+-expressing subsets. Compared with uninflamed, inflamed mucosa contained a marked increase in the proportion of the CD14hiHLA-DRdim cellular subset. This subset resembled the classical blood monocytes with low CD16, HLA-DR, and CX3CR1 expression. Classical monocytes migrated efficiently toward CCL2 and released the highest levels of MMP-1 and proinflammatory cytokines when stimulated with immune complexes or LPS. Our findings strongly suggest that it is the classical and not the intermediate or nonclassical monocytes that are the precursors to the dominating intestinal CD14hiHLA-DRdim subset. This enhances our understanding of CD pathology and may provide new options in treatment.
Technical Advance: A new cell preparation strategy that greatly improves the yield of vital and functional Tregs and NKT cells
Authors: Rissiek Björn , Danquah Welbeck , Haag Friedrich , Koch-Nolte Friedrich
Published: November 2013

Release of NAD+ during preparation of murine lymphocytes causes enzymatic ADP-ribosylation of cell-surface proteins on T cells, catalyzed by toxin-related ecto-ADP-ribosyltransferase, ARTC2. ADP-riboslyation activates the cytolytic P2X7 ion channel and affects, in particular, the vitality and function of Tregs and NKT cells. Here, we describe a simple method—injection of an ARTC2-blocking nanobody—to greatly improve Treg and NKT cell vitality and to preserve their function during in vitro assays and in adoptive-transfer experiments. Moreover, we present a method for the sorting of functional, primary NKT cells, based on coexpression of ARTC2 and NK1.1. Our results pave the way for the efficient ex vivo proliferation of Tregs and NKT cells and for new experimental and therapeutic uses of these important regulatory cells.
Migratory and adhesive properties of Xenopus laevis primordial germ cells in vitro
Authors: Dzementsei Aliaksandr , Schneider David , Janshoff Andreas , Pieler Tomas
Published: November 2013

The directional migration of primordial germ cells (PGCs) to the site of gonad formation is an advantageous model system to study cell motility. The embryonic development of PGCs has been investigated in different animal species, including mice, zebrafish, Xenopus and Drosophila. In this study we focus on the physical properties of Xenopus laevis PGCs during their transition from the passive to the active migratory state. Pre-migratory PGCs from Xenopus laevis embryos at developmental stages 17–19 to be compared with migratory PGCs from stages 28–30 were isolated and characterized in respect to motility and adhesive properties. Using single-cell force spectroscopy, we observed a decline in adhesiveness of PGCs upon reaching the migratory state, as defined by decreased attachment to extracellular matrix components like fibronectin, and a reduced adhesion to somatic endodermal cells. Data obtained from qPCR analysis with isolated PGCs reveal that down-regulation of E-cadherin might contribute to this weakening of cell-cell adhesion. Interestingly, however, using an in vitro migration assay, we found that movement of X. laevis PGCs can also occur independently of specific interactions with their neighboring cells. The reduction of cellular adhesion during PGC development is accompanied by enhanced cellular motility, as reflected in increased formation of bleb-like protrusions and inferred from electric cell-substrate impedance sensing (ECIS) as well as time-lapse image analysis. Temporal alterations in cell shape, including contraction and expansion of the cellular body, reveal a higher degree of cellular dynamics for the migratory PGCs in vitro.
Maintenance of muscle myosin levels in adult C. elegans requires both the double bromodomain protein BET-1 and sumoylation
Authors: Fisher Kate , Gee Fiona , Wang Siyao , Xue Feng , Knapp Stefan , Philpott Martin , Wells Christopher , Rodriguez Miriam , Snoek L. Basten , Kammenga Jan , Poulin Gino B.
Published: November 2013

Attenuation of RAS-mediated signalling is a conserved process essential to control cell proliferation, differentiation, and apoptosis. Cooperative interactions between histone modifications such as acetylation, methylation and sumoylation are crucial for proper attenuation in C. elegans, implying that the proteins recognising these histone modifications could also play an important role in attenuation of RAS-mediated signalling. We sought to systematically identify these proteins and found BET-1. BET-1 is a conserved double bromodomain protein that recognises acetyl-lysines on histone tails and maintains the stable fate of various lineages. Unexpectedly, adults lacking both BET-1 and SUMO-1 are depleted of muscle myosin, an essential component of myofibrils. We also show that this muscle myosin depletion does not occur in all animals at a specific time, but rather that the penetrance of the phenotype increases with age. To gain mechanistic insights into this process, we sought to delay the occurrence of the muscle myosin depletion phenotype and found that it requires caspase activity and MEK-dependent signalling. We also performed transcription profiling on these mutants and found an up-regulation of the FGF receptor, egl-15, a tyrosine kinase receptor acting upstream of MEK. Consistent with a MEK requirement, we could delay the muscle phenotype by systemic or hypodermal knock down of egl-15. Thus, this work uncovered a caspase- and MEK-dependent mechanism that acts specifically on ageing adults to maintain the appropriate net level of muscle myosin.
Does an instrumented treadmill correctly measure the ground reaction forces?
Authors: Willems Patrick A. , Gosseye Thierry P.
Published: November 2013

Since the 1990s, treadmills have been equipped with multi-axis force transducers to measure the three components of the ground reaction forces during walking and running. These measurements are correctly performed if the whole treadmill (including the motor) is mounted on the transducers. In this case, the acceleration of the treadmill centre of mass relative to the reference frame of the laboratory is nil. The external forces exerted on one side of the treadmill are thus equal in magnitude and opposite in direction to the external forces exerted on the other side. However, uncertainty exists about the accuracy of these measures: due to friction between the belt and the tread-surface, due to the motor pulling the belt, some believe that it is not possible to correctly measure the horizontal components of the forces exerted by the feet on the belt. Here, we propose a simple model of an instrumented treadmill and we demonstrate (1) that the forces exerted by the subject moving on the upper part of the treadmill are accurately transmitted to the transducers placed under it and (2) that all internal forces – including friction – between the parts of the treadmill are cancelling each other.