Evaluation of laboratory-based measures of soil mineral nitrogen and potentially mineralizable nitrogen as predictors of field-based indices of soil nitrogen supply in potato production

Accurate estimation of soil nitrogen (N) supply in the field is required to optimize fertilizer N management and to minimize environmental N losses in humid environments. Laboratory-based measures of N availability were evaluated as predictors of field-based indices of soil N supply within potato production systems. Pre-plant soil samples (0–15 cm) were collected from a series of forty treatments in established potato trials located in Atlantic Canada and Maine, USA. Total plant N uptake at topkill with no fertilizer N applied (PNU_0N), PNU_0N plus soil mineral N to 30 cm depth at harvest and relative yield were considered as field-based indices of soil N supply. The potentially mineralizable N (N₀) was determined by aerobic incubation at 25°C and periodic leaching for 24 weeks. A series of laboratory-based measures of soil N supply were measured in soil samples. Pre-plant soil nitrate or total mineral N at 0–30 cm depth was the best single predictor of PNU_0N (r = 0.67 and 0.71, respectively) and relative yield (r = 0.58 and 0.61). The ultraviolet absorbance of 0.01 M NaHCO₃ extract at 205 nm (NaHCO₃-205) was suitable as a predictor of PNU_0N and relative yield in each growing season, however, the relationship between this parameter and PNU_0N or relative yield varied somewhat among years. A combination of pre-plant mineral N plus N mineralized in the first 2 weeks period of incubation after re-wetting is proposed as a more robust measure of N availability compared with use of mineral N alone.     

Antileishmanial and Cytotoxic Effects of Essential Oil and Methanolic Extract of Myrtus communis L.

Plants used for traditional medicine contain a wide range of substances that can be used to treat various diseases such as infectious diseases. The present study was designed to evaluate the antileishmanial effects of the essential oil and methanolic extract of Myrtus communis against Leishmania tropica on an in vitro model. Antileishmanial effects of essential oil and methanolic extract of M. communis on promastigote forms and their cytotoxic activities against J774 cells were evaluated using MTT assay for 72 hr. In addition, their leishmanicidal activity against amastigote forms was determined in a macrophage model, for 72 hr. Findings showed that the main components of essential oil were α-pinene (24.7%), 1,8-cineole (19.6%), and linalool (12.6%). Findings demonstrated that M. communis, particularly its essential oil, significantly (P<0.05) inhibited the growth rate of promastigote and amastigote forms of L. tropica based on a dose-dependent response. The IC₅₀ values for essential oil and methanolic extract was 8.4 and 28.9 μg/ml against promastigotes, respectively. These values were 11.6 and 40.8 μg/ml against amastigote forms, respectively. Glucantime as control drug also revealed IC₅₀ values of 88.3 and 44.6 μg/ml for promastigotes and amastigotes of L. tropica, respectively. The in vitro assay demonstrated no significant cytotoxicity in J774 cells. However, essential oil indicated a more cytotoxic effect as compared with the methanolic extract of M. communis. The findings of the present study demonstrated that M. communis might be a natural source for production of a new leishmanicidal agent.     

Molecular pathogenesis of sialic acid storage diseases: insight gained from four missense mutations and a putative polymorphism of human sialin

Background information. Free sialic acid storage diseases are caused by mutations of a lysosomal sialic acid transporter called sialin. We showed recently that the milder clinical form, Salla disease, and a related non‐Finish case, are characterized by residual transport, whereas sialin mutants found in lethal infantile cases are inactive. In the present study, we have characterized the molecular effects of a putative polymorphism (M316I) and of four pathogenic mutations associated with either infantile (G127E and R57C) or Salla‐like (G409E) phenotypes, or both (G328E). The transport activity of human sialin was analysed using a novel assay that was based on a construct without the functional lysosomal sorting motif, which is expressed at the plasma membrane. Results. The lysosomal localization of human sialin was not (M316I and G328E) or only partially (R57C, G127E and G409E) affected by the missense mutations. In contrast, all pathogenic mutations abolished transport, whereas the putative M316I polymorphism induced an approx. 5‐fold decrease of sialic acid transport. Conclusions. The molecular effects of the R57C and G127E mutations strengthen the conclusion that the infantile phenotype is caused by loss‐of‐function mutations. On the other hand, the milder severity of the heterozygous G409E patient may reflect an incomplete expression of the splicing mutation present on the second allele. In the case of the G328E mutation, found in the homozygous state in a clinically heterogeneous family, the apparent severity of the transport phenotype suggests that the genetic or environmental factors underlying this clinical heterogeneity might be protective.     

Efficient phenotypic sex classification of zebrafish using machine learning methods

Sex determination in zebrafish by manual approaches according to current guidelines relies on human observation. These guidelines for sex recognition have proven to be subjective and highly labor‐intensive. To address this problem, we present a methodology to automatically classify the phenotypic sex using two machine learning methods: Deep Convolutional Neural Networks (DCNNs) based on the whole fish appearance and Support Vector Machine (SVM) based on caudal fin coloration. Machine learning techniques in sex classification provide potential efficiency with the advantage of automatization and robustness in the prediction process. Furthermore, since developmental plasticity can be influenced by environmental conditions, we have investigated the impact of elevated water temperature during embryogenesis on sex and sex‐related differences in color intensity of adult zebrafish. The estimated color intensity based on SVM was then applied to detect the association between coloration and body weight and length. Phenotypic sex classifications using machine learning methods resulted in a high degree of association with the real sex in nontreated animals. In temperature‐induced animals, DCNNs reached a performance of 100%, whereas 20% of males were misclassified using SVM due to a lower color intensity. Furthermore, a positive association between color intensity and body weight and length was observed in males. Our study demonstrates that high ambient temperature leads to a lower color intensity in male animals and a positive association of male caudal fin coloration with body weight and length, which appears to play a significant role in sexual attraction. The software developed for sex classification in this study is readily applicable to other species with sex‐linked visible phenotypic differences.     

Evaluation of fluorescent pseudomonads for plant growth promotion,antifungal activity against Rhizoctonia solani on common bean,and biocontrol potential

Fluorescent pseudomonads are ubiquitous bacteria that are common inhabitants of the rhizosphere and are the most studied group within the genus Pseudomonas. Bacterial isolates (n = 103) from the rhizosphere of wheat and common bean were assessed as potential biocontrol agents in this study. Fungal inhibition tests were performed by a plate assay in which each isolate was tested directly for the production of hydrogen cyanide, protease, siderophore and cellulase. Production of DAPG was verified by using an analytical high performance liquid chromatography assay (HPLC). Plant growth promotion was assessed in phytochamber trials and biocontrol activity was evaluated in greenhouse trials. In all, 52 bacterial isolates with antifungal activity against Rhizoctonia solani were found. Of the 52 isolates, 41 were selected according to their high efficiency in in vitro antagonism, which was shown as inhibition zones in the dual-culture assay. Six of the 41 rhizobacteria, including isolates UTPF7, UTPF13, UTPF18, UTPF22, UTPF27 and strain CHA0 produced HCN. Production of protease enzyme was detected for all isolates excluding UTPF30 isolate. Although some stains appeared not to produce any compound with affinity for ferric iron, other isolates produced prolific amounts, creating a large zone of orange (up to 160 mm², i.e., UTPF16). Seventeen of 41 isolates of fluorescent pseudomonads including strain CHAO produced different amounts of DAPG ranging from 0.6 to 11.4 ng/10⁸ cfu. A total of 39 isolates induced statistically significant effects on plant growth compared with the non-treated control for at least one parameter. The predominant influence observed was increased root length. No bacteria could completely protect the plant against R. solani, although all isolates significantly increased fresh weight as compared to the infested control in greenhouse trials. Pseudomonas fluorescens isolates UTPF16 and UTPF26 significantly (P < 0.05) decreased the number of seedlings with damping-off symptoms in the means of the experiments.     

Vaccines for colorectal cancer: an update

Colorectal cancer (CRC) is known as the third most common and fourth leading cancer associated death worldwide. The occurrence of metastasis has remained as a critical challenge in CRC, so that distant metastasis (mostly to the liver) has been manifested in about 20%-25% of patients. Several screening approaches have introduced for detecting CRC in different stages particularly in early stages. The standard treatments for CRC are surgery, chemotherapy and radiotherapy, in alone or combination. Immunotherapy is a set of novel approaches with the aim of remodeling the immune system battle with metastatic cancer cells, such as immunomodulatory monoclonal antibodies (immune checkpoint inhibitors), adoptive cell transfer (ACT) and cancer vaccine. Cancer vaccines are designed to trigger the intense response of immune system to tumor-specific antigens. In two last decades, introduction of new cancer vaccines and designing several clinical trials with vaccine therapy, have been taken into consideration in colon cancer patients. This review will describe the treatment approaches with the special attention to vaccines applied to treat colorectal cancer.     

In silico analysis of six known Leishmania major antigens and in vitro evaluation of specific epitopes eliciting HLA-A2 restricted CD8 T cell response

Background

As a potent CD8⁺ T cell activator, peptide vaccine has found its way in vaccine development against intracellular infections and cancer, but not against leishmaniasis. The first step toward a peptide vaccine is epitope mapping of different proteins according to the most frequent HLA types in a population.

Methods and Findings

Six Leishmania (L.) major-related candidate antigens (CPB,CPC,LmsTI-1,TSA,LeIF and LPG-3) were screened for potential CD8⁺ T cell activating 9-mer epitopes presented by HLA-A*0201 (the most frequent HLA-A allele). Online software including SYFPEITHI, BIMAS, EpiJen, Rankpep, nHLApred, NetCTL and Multipred were used. Peptides were selected only if predicted by almost all programs, according to their predictive scores. Pan-A2 presentation of selected peptides was confirmed by NetMHCPan1.1. Selected peptides were pooled in four peptide groups and the immunogenicity was evaluated by in vitro stimulation and intracellular cytokine assay of PBMCs from HLA-A2⁺ individuals recovered from L. major. HLA-A2⁻ individuals recovered from L. major and HLA-A2⁺ healthy donors were included as control groups. Individual response of HLA-A2⁺ recovered volunteers as percent of CD8⁺/IFN-γ⁺ T cells after in vitro stimulation against peptide pools II and IV was notably higher than that of HLA-A2⁻ recovered individuals. Based on cutoff scores calculated from the response of HLA-A2⁻ recovered individuals, 31.6% and 13.3% of HLA-A2⁺ recovered persons responded above cutoff in pools II and IV, respectively. ELISpot and ELISA results confirmed flow cytometry analysis. The response of HLA-A2⁻ recovered individuals against peptide pools I and III was detected similar and even higher than HLA-A2⁺ recovered individuals.

Conclusion

Using in silico prediction we demonstrated specific response to LmsTI-1 (pool II) and LPG-3- (pool IV) related peptides specifically presented in HLA-A*0201 context. This is among the very few reports mapping L. major epitopes for human HLA types. Studies like this will speed up polytope vaccine idea towards leishmaniasis.