Comparative physiological and transcriptome analysis in cultivated and wild sugarcane species in response to hydrogen peroxide-induced oxidative stress.

BACKGROUND: Sugarcane is an important energy crop grown worldwide,supplementing various renewable energy sources. Cultivated and wild sugarcane species respond differently to biotic and abiotic stresses. Generally, wild species are tolerant to various abiotic stresses. In the present study, the physiological and molecular responses of cultivated and wild sugarcane species to oxidative stress at the transcriptional levels were compared. Transcriptional responses were determined using RNAseq. The representative RNA-seq transcript values were validated by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) and confirmed through physiological responses. RESULTS: Oxidative stress causes leaf-rolling and -tip drying in cultivated sugarcane, but the wild species are tolerant. Higher chlorophyll fluorescence was observed in the wild species than that in the cultivated varieties under stress. Wild species can maintain a higher chlorophyll stability index than the cultivated species, which was confirmed by the lower transcripts of the chlorophyllase gene in the wild species than that in the cultivated variety. Transcription factor genes (NAC, MYB, and WRKY) were markedly expressed in response to oxidative stress, revealing their involvement in stress tolerance. The analysis revealed synchronized expression of acetyl-transferase, histone2A, cellulose synthase, and secondary cell wall biosynthetic genes in the wild species. The validation of selected genes and 15 NAC transcription factors using RT-qPCR revealed that their expression profiles were strongly correlated with RNA-seq. To the best of our knowledge, this is the first report on the oxidative stress response in cultivated and wild sugarcane species. CONCLUSION: Physiological and biochemical changes in response to oxidative stress markedly differ between cultivated and wild sugarcane species. The differentially expressed stress-responsive genes are grouped intothe response to oxidative stress, heme-binding, peroxidase activity, and metal ion binding categories. Chlorophyll maintenance is a stress tolerance response enhanced by the differential regulation of the chlorophyllase gene.There is a considerable difference in the chlorophyll stability index between wild and cultivated varieties. We observed a substantial regulation of secondary wall biosynthesis genes in the wild species compared with that in the cultivated variety, suggesting differences in stress tolerance mechanisms.

Doppler assessment of the fetus in pregnant women recovered from COVID-19.

OBJECTIVE: The aim of this study was to evaluate the maternal-fetal Doppler patterns in pregnant women recovered from COVID-19. METHODS: This prospective case-control study was conducted in Ankara City Hospital between July 1, 2020 and August 30, 2020. Thirty pregnant women who were diagnosed with COVID-19 and completed the quarantine process were compared with 40 healthy pregnant women in terms of the fetal Doppler parameters. All pregnant women diagnosed with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection were followed up in our clinic and their diagnoses have been confirmed in nasopharyngeal and oropharyngeal samples by quantitative real time reverse transcriptase polymerase chain reaction (RT-PCR) method. Doppler ultrasonographic assessment of the uterine arteries (UtA) and middle cerebral artery (MCA) were used in addition to umbilical artery (UA) Doppler between 23 and 40 weeks of gestation. Also, cerebroplacental ratio (CPR) was calculated according to gestational age. RESULTS: The pulsatility and resistance indices of umbilical and UtA showed a significant increase in pregnant women in the study group compared to the control group (p < 0.05). Multivariable logistic regression analysis revealed that pulsatility and resistance indices of the mean UtA were independently associated with disease (OR > 1000, 95%CI 9.77 to >1000, p = 0.009; OR 0,000 95%CI 0,000-0,944, p = 0,049), respectively. Medical treatment was given to 16/30 (53%) of pregnant women diagnosed with COVID-19. CONCLUSION: In conclusion, uterine artery Doppler indices in the third trimester may have clinical value in pregnant women recovered from COVID-19.

Comparative analysis of drought-responsive transcriptomes of sugarcane genotypes with differential tolerance to drought.

Water stress causes considerable yield losses in sugarcane. To investigate differentially expressed genes under water stress, two sugarcane genotypes were subjected to three water-deficit levels (mild, moderate, and severe) and subsequent recovery and leaf transcriptome was generated using Illumina NextSeq sequencing. Among the differentially expressed genes, the tolerant genotype Co 06022 generated 2970 unigenes (p ≤ 0.05, functionally known, non-redundant DEGs) at 2-day stress, and there was a progressive decrease in the expressed genes as the stress period increased with 2109 unigenes at 6-day stress and 2307 unigenes at 10-day stress. There was considerable reduction at recovery with 1334 unigenes expressed at 10 days after recovery. However, in the susceptible genotype Co 8021, the number of unigenes expressed at 2 days was lower (2025) than the tolerant genotype and a further reduction was seen at 6-day stress (1552). During recovery, more differentially expressed genes were observed in the susceptible cultivar indicating that the cultivar has to activate more functions/processes to recover from the damage caused by stress. Comparison of DEGs between all stages of stress and recovery in both genotypes revealed that, the commonly up- and down-regulated genes across different stages were approximately double in the tolerant genotype. The most enriched gene ontology classes were heme binding, peroxidase activity and metal ion binding in the biological process and response to oxidative stress, hydrogen peroxide catabolic process and response to stress in the molecular function category. The cellular component was enriched with DEGs involved in extracellular region followed by integral component of membrane. The KEGG pathway analysis revealed important metabolic activities and functionally important genes involved in mitigating water-deficit stress in both the varieties. In addition, several unannotated genes in important pathways were detected and together may provide novel insights into water-deficit tolerance mechanisms in sugarcane. The reliability of the observed expression patterns was confirmed by qRT-PCR. The results of this study will help to identify useful genes for improving drought tolerance in sugarcane.

Brain Tumor Segmentation Using Convolutional Neural Networks in MRI Images.

In medical image processing, Brain tumor segmentation plays an important role. Early detection of these tumors is highly required to give Treatment of patients. The patient's life chances are improved by the early detection of it. The process of diagnosing the brain tumoursby the physicians is normally carried out using a manual way of segmentation. It is time consuming and a difficult one. To solve these problems, Enhanced Convolutional Neural Networks (ECNN) is proposed with loss function optimization by BAT algorithm for automatic segmentation method. The primary aim is to present optimization based MRIs image segmentation. Small kernels allow the design in a deep architecture. It has a positive consequence with respect to overfitting provided the lesser weights are assigned to the network. Skull stripping and image enhancement algorithms are used for pre-processing. The experimental result shows the better performance while comparing with the existing methods. The compared parameters are precision, recall and accuracy. In future, different selecting schemes can be adopted to improve the accuracy.

Potentiality of yeast Candida sp. SMN04 for degradation of cefdinir, a cephalosporin antibiotic: kinetics, enzyme analysis and biodegradation pathway.

A new yeast strain isolated from the pharmaceutical wastewater was capable of utilizing cefdinir as a sole carbon source for their growth in mineral medium. The yeast was identified and named as Candida sp. SMN04 based on morphology and 18S-ITS-D1/D2/D3 rRNA sequence analysis. The interaction between factors pH (3.0-9.0), inoculum dosage (1-7%), time (1-11 day) and cefdinir concentration (50-450 mg/L) was studied using a Box-Behnken design. The factors were studied as a result of their effect on cell dry weight (R1; g/L), extended spectrum ß-lactamase (ESBL) assay (R2; mm), P450 activity (R3; U/mL) and degradation (R4; %). Maximum values of R1, R2, R3 and R4 were obtained at central values of all the parameters. The isolated yeast strain efficiently degraded 84% of 250 mg L⁻¹ of cefdinir within 6 days with a half-life of 2.97 days and degradation rate constant of 0.2335 per day. Pseudo-first-order model efficiently described the process. Among the various enzymes tested, the order of activity at the end of Day 4 was noted to be: cytochrome P450 (1.76 ± 0.03) > NADPH reductase (1.51 ± 0.20) > manganese peroxidase and amylase (0.66 ± 0.15; 0.66 ± 0.70). Intermediates were successfully characterized by liquid chromatography-mass spectrometry. The opening of the ß-lactam ring involving ESBL activity was considered as one of the major steps in the cefdinir degradation process. Fourier transform-infrared spectroscopy analysis showed the absence of spectral vibrations between 1766 and 1519 cm⁻¹ confirming the complete removal of lactam ring during cefdinir degradation. The results of the present study are promising for the use of isolated yeast Candida sp. SMN04 as a potential bioremediation agent.

Purification and characterization of carbon-phosphorus bond-cleavage enzyme from glyphosate degrading Pseudomonas putida T5.

An inducible, carbon-phosphorus bond-cleavage enzyme was purified from cells of Pseudomonas putida T5 grown on N-phosphonomethyl glycine. The native enzyme had a molecular mass of approximately 70 kD and upon sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), yielded a homogeneous protein band with an apparent molecular mass of about 70 kD. Activity of purified enzyme was increased by 627-fold compared to the crude extract and showed pH and temperature optima of approximately 7 and 30°C, respectively. The purified enzyme had an apparent Km and Vmax of 3.7 mM and 6.8 mM/min, respectively, for its sole substrate N-phosphonomethyl glycine. The enzyme was inhibited by phenylmethylsulfonyl fluoride (PMSF), indicating the presence of serine at the active site. The enzyme was not inhibited by SDS, suggesting the absence of disulfide linkage in the enzyme. The enzyme was found to be inhibited by most of the metals studied except Mg(2+). Detergents studied also inhibited glyphosate acting as a carbon-phosphorus bond-cleavage enzyme. Thus initial characterization of the purified enzyme suggested that it could be used as a potential candidate for glyphosate bioremediation.

Biodegradation of cefdinir by a novel yeast strain, Ustilago sp. SMN03 isolated from pharmaceutical wastewater.

Cefdinir, a semi-synthetic third generation cephalosporin antibiotic being considered as an emerging pollutant, demands removal from aquatic ecosystems. A yeast strain isolated from pharmaceutical wastewater which was identified as Ustilago sp. SMN03 by molecular techniques and was found to be capable of utilizing cefdinir as a sole carbon source. The isolate was found to degrade 81 % of cefdinir within 6 days under optimized conditions viz. pH 6.0, temperature 30 °C, a shaking speed of 120 rpm, an inoculum dosage of 4 % (w/v) and an initial cefdinir concentration of 200 mg L(-1). Kinetic studies revealed that cefdinir degradation followed the pseudo-first order model, a rate constant of 0.222 per day and a half-life period of 3.26 days. Using LC-MS analysis, six novel intermediates formed during the cefdinir degradation were identified and characterized. FT-IR analysis showed that the functional groups ranging from 1,766 to 1,519 cm(-1), characteristic for lactam ring were completely removed during the cefdinir degradation. The opening of the ß-lactam ring was one of the major steps in the cefdinir degradation process. Based on the results from the present study, a possible pathway of cefdinir degradation by Ustilago sp. SMN03 was proposed. To the best of our knowledge, this is the first report on microbial degradation of cefdinir by yeast.

Detection of isoprothiolane in food, soil, and water samples by immunosorbent assay using avian antibodies.

A simple competitive immunoassay was developed for the measurement of isoprothiolane in rice, soil, and water samples. It employed the avian antibodies (IgY) that recognized isoprothiolane as a capture reagent and isoprothiolane-alkaline phosphatase conjugate as an enzyme label. The assay depended on the competitive binding between the anti-isoprothiolane antibody and isoprothiolane derived from rice, soil, and water samples for binding sites with immobilized isoprothiolane-ovalbumin (OVA) conjugate. The concentration of isoprothiolane in the rice, soil, and water samples was quantified by the ability of the pesticide present in the samples to inhibit the binding of the enzyme conjugate to the antibody and subsequently the color formation in the assay. The assay was specific to isoprothiolane with a limit of detection of 2 ng/mL. Mean analytical recovery of isoprothiolane in different rice matrices was 87.20%-98.02%, for soil samples recovery was 74.24%-111.20%, and water samples recovery was 35.2%-95.73%. The precision of the assay was satisfactory. The assay compared favorably with gas chromatography (GC) in its ability to accurately measure isoprothiolane in the different rice, soil, and water samples.

Degradation of isoprothiolane by a defined microbial consortium using response surface methodology.

A defined microbial consortium was developed for the degradation of isoprothiolane. Isoprothiolane-biodegradation parameters were optimized using Response Surface Methodology (RSM). Three variables chosen for the study were inoculum concentration (50-1500 microg protein ml(-1)), temperature (25-35 degrees C) and pH (4-8) each at levels -1.682, -1, 0, 1 and 1.682. Incubation time of 72 hr was kept constant. Degradation of different concentrations of isoprothiolane was studied. The optimized conditions obtained were, inoculum concentration of 50 microg protein ml(-1) at 30 degrees C and pH between 4-8. The maximum predicted percentage degradation of 100, 100, 100, 100 and 95.5 was obtained respectively for 5, 10, 20, 30 and 50 ppm of initial isoprothiolane concentrations at different pH levels 7.7, 6.8, 6.2, 4.7 and 4.6. Validation of the model indicated that experimental values were found to be in agreement with the predicted one.

[Prevention of psychosocial issues and drug addiction in workers employed in building great infrastructures: first outcomes of an experience in the Florence area]. / La prevenzione del disagio psicosociale lavorativo e delle dipendenze negli addetti alla realizzazione delle grandi opere infrastrutturali: primi risultati di uno esperienza in territorio fiorentino.

The research on stress, mobbing, and substance dependence in workers employed in the building of the great infrastructures is part of the project Euridice-Ten. The 75% of the workers employed took part in the research through an anonymous and structured questionnaire. The Clinica del Lavoro of the University of Milan elaborated the data. They point out a high risk of stress, a low level of control, a marked risk of mobbing, a high chronic fatigue in the most exhausting jobs, sleep disorders, problems in the social life as well as at home, in nourishment and at work. Many workers drink too much alcohol and the 9% of the workers know workers who use cocaine.

Role of magnetic resonance imaging in evaluating sphenoid sinus and internal carotid artery.

OBJECTIVE: This study aimed to determine whether magnetic resonance imaging has a role in the evaluation of the sphenoid sinus and internal carotid artery. In addition, we aimed to establish reference measurements for the minimal distance between the internal carotid arteries. METHOD: The sphenoid sinuses and neighbouring internal carotid arteries of 90 patients were evaluated using sagittal T1-weighted and axial and coronal T2-weighted magnetic resonance images. RESULTS: Sphenoid sinus pneumatisation was categorised as occipitosphenoidal (0 per cent), conchal (3.3 per cent), presellar (14.4 per cent) or sellar (82.2 per cent). The internal carotid artery protruded into the sphenoid sinus in 32.8 per cent, with a septum in 9.4 per cent. The incidence of sellar-type sphenoid sinus pneumatisation was higher in patients with protrusion of the internal carotid artery into the sphenoid sinus (p < 0.001). The incidence of presellar pneumatisation was higher in patients without internal carotid artery protrusion (p < 0.001). The minimal distance between the internal carotid arteries varied between 9.04 and 24.26 mm (mean, 15.94 mm). CONCLUSION: Magnetic resonance imaging can provide useful information about the sphenoid sinus and internal carotid artery, prior to endoscopic sphenoidotomy and trans-sphenoidal hypophysectomy.

A successful management of dengue fever in pregnancy: report of two cases.

In the most recent outbreak of dengue fever in India, we encountered at least seven cases of fever with thrombocytopenia in pregnancy but only two were seropositive for dengue. In one of the cases there was postpartum hemorrhage while in the other case there was perinatal transmission to the neonate requiring platelet transfusions. The diagnostic difficulties in pregnancy and the management are discussed.

Effect of cyclosporin A and tacrolimus on sister chromatid exchange frequency in renal transplant patients.

Long-term use of Cyclosporin A (CsA) and Tacrolimus is known to yield serious untoward side effects including nephrotoxicity, neurotoxicity, and malignant tumor formation. Sister chromatid exchange (SCE) is used to assess the genotoxic potential of various agents. A total of 37 postrenal transplant patients receiving either CsA (n = 20) or Tacrolimus (n = 17) were included in this study. The genotoxic effects of CsA and Tacrolimus were assessed by determination of SCE frequency. In patients receiving CsA, SCE frequency was increased significantly compared to that in the control group (p = 0.001), whereas Tacrolimus did not yield such a significant change (p = 0.801). SCE frequency was not correlated with drug dosage (p > 0.05). Our results indicate that the use of CsA, but not Tacrolimus 506, is associated with an increased genotoxic effect in postrenal transplant patients.

Quantitative and diffusion MR imaging as a new method to assess osteoporosis.

BACKGROUND AND PURPOSE: The role of advanced MR imaging techniques in demonstrating the microarchitectural changes in osteoporosis has been recently investigated. Our aim was to determine the role of quantitative and diffusion MR imaging in the diagnosis of osteoporosis compared with dual-energy x-ray absorptiometry (DEXA). MATERIALS AND METHODS: Fifty-one subjects underwent both DEXA and conventional MR imaging with diffusion and apparent diffusion coefficient (ADC) mapping. T1, T2, diffusion signal intensity, and ADC values were calculated and compared with bone mineral attenuation (BMD). Results were obtained from L1 to L4 of each patient with a total of 204 lumbar vertebrae. RESULTS: Both T1 and T2 signal intensities tended to increase when both diffusion and ADC values decreased with reducing BMD. CONCLUSIONS: An inverse relationship between BMD and T1 and T2 signal intensities and a direct relationship between diffusion and ADC values were present in this pilot group. The T1/diffusion signal-intensity ratio could be a reliable diagnostic indicator of osteoporosis.

Partial purification and characterization of protease of Bacillus proteolyticus CFR3001 isolated from fish processing waste and its antibacterial activities.

Bacillus proteolyticus CFR3001 isolated from fish processing wastes (both fresh water and marine) produced an alkaline protease. The optimum conditions for cell growth and protease production were 37 degrees C, 96 h, agitation speed of 100 rpm and medium pH 9. The partially purified protease obtained from had specific activity of 22.05 at 37 degrees C was active between 40 degrees C and 50 degrees C and lost >20% of its activity around 60 degrees C. Its molecular weight was approximately 29 kDa and it inhibited the growth of several pathogenic organisms such as Escherichia coli, Listeria monocytogenes, Bacillus cereus and Yersinia enterocolytica. The scanning electron microscopy (SEM) studies revealed that the protease produced by B. proteolyticus CFR3001 lysed the cells of these pathogenic bacteria.