Droplet digital polymerase chain reaction assay and peptide nucleic acid-locked nucleic acid clamp method for RHOA mutation detection in angioimmunoblastic T-cell lymphoma.

Angioimmunoblastic T-cell lymphoma (AITL) is a subtype of nodal peripheral T-cell lymphoma (PTCL). Somatic RHOA mutations, most frequently found at the hotspot site c.50G > T, p.Gly17Val (G17V RHOA mutation) are a genetic hallmark of AITL. Detection of the G17V RHOA mutations assists prompt and appropriate diagnosis of AITL. However, an optimal detection method for the G17V RHOA mutation remains to be elucidated. We compared the sensitivity and concordance of next-generation sequencing (NGS), droplet digital PCR (ddPCR) and peptide nucleic acid-locked nucleic acid (PNA-LNA) clamp method for detecting the G17V RHOA mutation. G17V RHOA mutations were identified in 27 of 67 (40.3%) PTCL samples using NGS. ddPCR and PNA-LNA clamp method both detected G17V mutations in 4 samples in addition to those detected with NGS (31 of 67, 46.3%). Additionally, variant allele frequencies with ddPCR and those with NGS showed high concordance (P < .001). Three other RHOA mutations involving the p.Gly17 position (c.[49G > T;50G > T], p.Gly17Leu in PTCL198; c.[50G > T;51A > C], p.Gly17Val in PTCL216; and c.50G > A, p.Gly17Glu in PTCL223) were detected using NGS. These sequence changes could not appropriately be detected using the ddPCR assay and the PNA-LNA clamp method although both indicated that the samples might have mutations. In total, 34 out of 67 PTCL samples (50.7%) had RHOA mutations at the p.Gly17 position. In conclusion, our results suggested that a combination of ddPCR/PNA-LNA clamp methods and NGS are best method to assist the diagnosis of AITL by detecting RHOA mutations at the p.Gly17 position.

Detection of the circulating tumor DNAs in angioimmunoblastic T- cell lymphoma.

Recent genetic studies identified that the disease-specific G17V RHOA mutation, together with mutations in TET2, DNMT3A, and IDH2, is a hallmark of angioimmunoblastic T cell lymphomas (AITL). The diagnostic value of these mutations is now being investigated. Circulating tumor DNAs (ctDNAs) may offer a non-invasive testing for diagnosis and disease monitoring of cancers. To investigate whether these mutations are useful markers for ctDNAs in AITL and its related lymphomas, we performed targeted sequencing for TET2, RHOA, DNMT3A, and IDH2 in paired tumors and cell-free DNAs from 14 patients at diagnosis. Eighty-three percent of mutations detected in tumors were also observed in cell-free DNAs. During the disease course, mutations were detectable in cell-free DNAs in a refractory case, while they disappeared in a chemosensitive case. These data suggest that the disease-specific gene mutations serve as sensitive indicators for ctDNAs and may also be applicable for non-invasive monitoring of minimal residual diseases in AITL.

Submerged anaerobic membrane bioreactor (SAnMBR) performance on sewage treatment: removal efficiencies, biogas production and membrane fouling.

A submerged anaerobic membrane reactor (SAnMBR) was employed for comprehensive evaluation of sewage treatment at 25 °C and its performance in removal efficiency, biogas production and membrane fouling. Average 89% methanogenic degradation efficiency as well as 90%, 94% and 96% removal of total chemical oxygen demand (TCOD), biochemical oxygen demand (BOD) and nonionic surfactant were obtained, while nitrogen and phosphorus were only subjected to small removals. Results suggest that SAnMBRs can effectively decouple organic degradation and nutrients disposal, and reserve all the nitrogen and phosphorus in the effluent for further possible recovery. Small biomass yields of 0.11 g mixed liquor volatile suspended solids (MLVSS)/gCOD were achieved, coupled to excellent methane production efficiencies of 0.338 NLCH4/gCOD, making SAnMBR an attractive technology characterized by low excess sludge production and high bioenergy recovery. Batch tests revealed the SAnMBR appeared to have the potential to bear a high food-to-microorganism ratio (F/M) of 1.54 gCOD/gMLVSS without any inhibition effect, and maximum methane production rate occurred at F/M 0.7 gCOD/gMLVSS. Pore blocking dominated the membrane fouling behaviour at a relative long hydraulic retention time (HRT), i.e. >12 hours, while cake layer dominated significantly at shorter HRTs, i.e. <8 hours.

Extension of Light-Harvesting Ability of Photosynthetic Light-Harvesting Complex 2 (LH2) through Ultrafast Energy Transfer from Covalently Attached Artificial Chromophores.

Introducing appropriate artificial components into natural biological systems could enrich the original functionality. To expand the available wavelength range of photosynthetic bacterial light-harvesting complex 2 (LH2 from Rhodopseudomonas acidophila 10050), artificial fluorescent dye (Alexa Fluor 647: A647) was covalently attached to N- and C-terminal Lys residues in LH2 α-polypeptides with a molar ratio of A647/LH2 ≃ 9/1. Fluorescence and transient absorption spectroscopies revealed that intracomplex energy transfer from A647 to intrinsic chromophores of LH2 (B850) occurs in a multiexponential manner, with time constants varying from 440 fs to 23 ps through direct and B800-mediated indirect pathways. Kinetic analyses suggested that B800 chromophores mediate faster energy transfer, and the mechanism was interpretable in terms of Förster theory. This study demonstrates that a simple attachment of external chromophores with a flexible linkage can enhance the light harvesting activity of LH2 without affecting inherent functions of energy transfer, and can achieve energy transfer in the subpicosecond range. Addition of external chromophores, thus, represents a useful methodology for construction of advanced hybrid light-harvesting systems that afford solar energy in the broad spectrum.

Possible factors determining global-scale patterns of crop yield sensitivity to drought.

In recent decades, droughts have critically limited crop production, inducing food system shocks regionally and globally. It was estimated that crop yield variability in around one-third to three-fourths of global harvested areas is explained significantly by drought, revealing the notable vulnerability of crop systems to such climate-related stressors. However, understanding the key factors determining the global pattern of crop yield sensitivity to drought is limited. Here, we investigate a wide range of physical and socioeconomic factors that may determine crop-drought vulnerability in terms of yield sensitivity to drought based on the Standardized Precipitation Index at 0.5° resolution from 1981 to 2016 using machine learning approaches. The results indicate that the spatial variations of the crop-drought sensitivity were mainly explained by environmental factors (i.e., annual precipitation, soil water-holding capacity, soil acidity, annual potential evapotranspiration) and crop management factors (i.e., fertilizer rate, growing season). Several factors might have a positive effect in mitigating crop-drought vulnerability, such as annual precipitation, soil water holding capacity, and fertilizer rate. This study quantitatively assesses the possible effect of various determinants which might control crop vulnerability to drought. This understanding may provide insights for further studies addressing better crop vulnerability measures under future drought stress.

Estimating monthly total nitrogen concentration in streams by using artificial neural network.

Artificial Neural Network (ANN) is a flexible and popular tool for predicting the non-linear behavior in the environmental system. Here, the feed-forward ANN model was used to investigate the relationship among the land use, fertilizer, and hydrometerological conditions in 59 river basins over Japan and then applied to estimate the monthly river total nitrogen concentration (TNC). It was shown by the sensitivity analysis, that precipitation, temperature, river discharge, forest area and urban area have high relationships with TNC. The ANN structure having eight inputs and one hidden layer with seven nodes gives the best estimate of TNC. The 1:1 scatter plots of predicted versus measured TNC were closely aligned and provided coefficients of errors of 0.98 and 0.93 for ANNs calibration and validation, respectively. From the results obtained, the ANN model gave satisfactory predictions of stream TNC and appears to be a useful tool for prediction of TNC in Japanese streams. It indicates that the ANN model was able to provide accurate estimates of nitrogen concentration in streams. Its application to such environmental data will encourage further studies on prediction of stream TNC in ungauged rivers and provide a useful tool for water resource and environment managers to obtain a quick preliminary assessment of TNC variations.

Effect of Cellulose Nanocrystals Extracted from Oil Palm Empty Fruit Bunch as Green Admixture for Mortar.

This paper aims to examine the effect of cellulose nanocrystals (CNCs) derived from oil palm empty fruit bunch fiber (EFB) incorporating cement mortar on its structural performances. Cellulose nanocrystals (CNCs) were extracted from α-cellulose extracted from EFB using an acid hydrolysis process with a concentration of acid used was 64% w/v under the temperature of 45 °C for 60 minutes. The Cellulose nanocrystals (CNCs) were mixed into the cement mortar ranging from 0 to 0.8% w/w and its mechanical properties were determined. The developed CNCs mortar was characterized for their compressive and flexural properties as well as microstructure. The influence of CNCs concentration, curing method, dispersion of CNCs on mortar's mechanical performance was thoroughly examined to find out the optimum condition. Overall results revealed that an addition of 0.4% cellulose nanocrystals has shown to increase the compressive and flexural strength to 46% and 20%, respectively cured under the wrapping method. The hydration of cementitious composites also improved significantly with the addition of CNCs by the formation of highly crystalline of portlandite observed under the XRD test. This present work demonstrates the importance of palm oil empty fruit bunch waste as a sustainable resource of cellulose nanocrystals admixture to achieve structural strength of cement mortar and promotes green technologies in construction.

Profile of regional fat and fat-free soft tissue accumulation in male athletes.

BACKGROUND: It is unclear whether or not the breakpoint (BP), at which the proportion of each of fat mass (FM) and fat-free soft tissue mass (FFSTM) to body mass (BM) alter, exists in male athletes. We examined the hypothesis that in male athletes, the regional FM and FFSTM-BM relationships have a BP, but the body mass at BP (BMBP) differs among the arms, trunk, and legs. METHODS: By using a dual X-ray absorptiometry, whole-body and regional FMs and FFSTMs in the arms, trunk, and legs were estimated in 198 male athletes (20.8 ± 2.1 years; 1.73 ± 0.07 m; 72.7 ± 14.8 kg). To detect the BP in the relationship between each of FM and FFSTM and BM, a piecewise linear regression analysis was used. If a BP was detected in the corresponding relationship, the significant difference between the regression slopes above and below the BP was examined. RESULTS: The regression analysis indicated that the BMBP existed in the FM- and FFSTM-BM relationships regardless of region and whole body. For the whole body, BMBP was 81.8 kg for FM and 82.2 kg for FFSTM. In regional FM-BM relationships, BMBP was 80.5 kg for arms, 82.6 kg for trunk, and 63.3 kg for legs, and the regression slopes above the BMBP became higher than those below the BP, and vice versa in regional FFSTM-BM relationships (BMBP 104.6 kg for arms, 80.9 kg for trunk, and 79.0 kg for legs). The relative differences in the slopes between below and above BMBP in the regional FM-BM relationships were higher in the arms and trunk than in the legs, and those in the regional FFSTM-BM relationships in the legs than in the trunk. CONCLUSION: Whole-body and regional FM- and FFSTM-BM relationships for male athletes have breakpoints at which the proportion of the tissue masses to BM alters. The BMBP and differences in the distribution of regional FM and FFSTM around the breakpoint are region specific.

TX99 Is a Neutralizing Monoclonal Antibody Against Mouse TIGIT.

T cell immunoglobulin and ITIM domains (TIGIT) is an inhibitory immunoreceptor expressed on NK cells, effector and memory T cells, and regulatory T cells (Tregs). The ligands for TIGIT are CD155 (PVR) and CD112 (PVRL2, nectin-2), which are broadly expressed on hematopoietic cells and nonhematopoietic cells. TIGIT negatively regulates antitumor responses, but promotes autoimmune reaction. Although neutralizing anti-human TIGIT mAbs are under clinical trials for cancers, how the blockade of TIGIT interaction with the ligands shows tumor immunity still remains unclear. Although analyses of mouse tumor model using a neutralizing anti-mouse TIGIT (mTIGIT) mAbs should be useful to address this issue, there are limitations to this type of studies due to unavailability of neutralizing anti-mTIGIT mAbs. In this study, we generated five clones of anti-mTIGIT mAbs, designated TX99, TX100, TX103, TX104, and TX105. We show that TX99 and TX100 showed the strongest binding to TIGIT. We also show that TX99 interfered with the interaction between TIGIT and CD155 and increased NK cell-mediated cytotoxicity against CD155-expressing RMA-S cells. Thus, TX99 is a unique neutralizing mAb that can be used for studies of mTIGIT functions.

Body shape indices are predictors for estimating fat-free mass in male athletes.

It is unknown whether body size and body shape parameters can be predictors for estimating whole body fat-free mass (FFM) in male athletes. This study aimed to investigate whether body size and shape variables can be predictors for FFM in male athletes. Using a whole-body dual-energy X-ray absorptiometry scanner, whole body fat mass (FM) and FFM were determined in 132 male athletes and 14 sedentary males. The sample was divided into two groups: validation (N = 98) and cross-validation (N = 48) groups. Body height (BH), body mass (BM), and waist circumference at immediately above the iliac crest (W) were measured. BM-to-W and W-to-BH ratios were calculated as indices of body shapes. Stepwise multiple regression analysis revealed that BM/W and W/BH were selected as explainable variables for predicting FFM. The equation developed in the validation group was FFM (kg) = 0.883 × BM/W (kg/m) + 43.674 × W/BH (cm/cm)- 41.480 [R2 = 0.900, SEE (%SEE) = 2.3 kg (3.8%)], which was validated in the cross-validation group. Thus, the current results demonstrate that an equation using BM/W and W/BH as independent variables is applicable for predicting FFM in male athletes.

Investigation on the response of anaerobic membrane bioreactor to temperature decrease from 25°C to 10°C in sewage treatment.

Anaerobic membrane bioreactor (AnMBR) for sewage treatment was operated for 650days with the decrease of temperature from 25°C to 10°C. At higher temperature >15°C, COD removal was above 94% while sewage treatment efficiency and relevant CH4 production decreased below 15°C. The effluent COD at 10°C was 134mg/L at HRT of 16h. Moreover, low temperature can result in a higher membrane fouling rate due to the microbial self-protection behavior in coping with the temperature decrease by releasing soluble microbial products (SMP) and extracellular polymeric substances (EPS). The contribution of pore blocking to membrane fouling caused by protein from SMP and EPS increased from 17% to 45% and that of cake layer decreased from 81% to 53% at 25°C and 15°C respectively. The inhibition to hydrolysis and acidification process was responsible to the decrease of sewage treatment at lower temperature.