Utjecaj temperature uzgoja na ekspresiju gena za biosintezu pigmenta monakolina K u plijesni Monascus sp.

In this study, the effects of temperature-shift (from 30 to 25 °C) and temperature-constant (at 30 °C) cultivation on the mass of Monascus fuliginosus CG-6 mycelia and concentration of the produced monacolin K (MK) were monitored. The expression levels of the MK biosynthetic genes of M. fuliginosus CG-6 at constant and variable culture temperatures were analysed by real-time quantitative polymerase chain reaction (RT-qPCR). The total protein was collected and determined by liquid chromatography-electrospray ionisation with tandem mass spectrometry (LC-ESI-MS/MS). Results showed that the maximum mycelial mass in temperature-shift cultivation was only 0.477 g of dry cell mass per dish, which was lower than that in temperature-constant cultivation (0.581 g of dry cell mass per dish); however, the maximum concentration of MK in temperature-shift cultivation (34.5 μg/mL) was 16 times higher than that in temperature-constant cultivation at 30 °C (2.11 μg/mL). Gene expression analysis showed that the expression of the MK biosynthetic gene cluster at culture temperature of 25 °C was higher than that at 30 °C, which was similar to the trend of the MK concentration, except for individual MK B and MK C genes. Analysis of differential protein expression revealed that 2016 proteins were detected by LC-ESI-MS/MS. The expression level of efflux pump protein coded by the MK I gene exhibited the same upregulated trend as the expression of MK I in temperature-shift cultivation. Temperature-shift cultivation enhanced the expression of proteins in the secondary metabolite production pathway, but suppressed the expression of proteins involved in the mycelial growth.U radu je istražen utjecaj promjene temperature (s 30 na 25 °C) i konstantne temperature uzgoja (30 °C) na masu micelija plijesni Monascus fuliginosus CG-6 i koncentraciju proizvedenog monakolina K, pri čemu je razina ekspresije gena za biosintezu monakolina praćena metodom RT-qPCR. Ukupni su proteini određeni metodom LC -ESI-MS/MS. Rezultati pokazuju da je maksimalna masa micelija dobivenog pri promjeni temperature uzgoja bila samo 0,477 g suhe tvari po Petrijevoj zdjelici, što je znatno manje od one dobivene uzgojem pri konstantnoj temperaturi (0,581 g suhe tvari po Petrijevoj zdjelici). Međutim, maksimalna koncentracija monakolina K pri promjeni temperature uzgoja bila je 34,5 μg/mL, tj. 16 puta veća od one dobivene uzgojem pri konstantnoj temperaturi (2,11 μg/mL). U skladu s tim, utvrđeno je da je razina ekspresije klastera gena za biosintezu monakolina K veća pri 25 nego pri 30 °C, osim za gene MK B i MK C. Analizom diferencijalne ekspresije proteina otkriveno je 2016 različitih proteina, identificiranih metodom LC-ESI-MS/MS. Ekspresija proteinske pumpe koju kodira MK I gen pokazala je isti trend kao i ekspresija samog gena pri promjeni temperature uzgoja s 30 na 25 °C. Promjena temperature uzgoja povećala je ekspresiju proteina uključenih u sintezu sekundarnih metabolita, no smanjila ekspresiju proteina odgovornih za rast micelija

Development of SCAR Marker Related to Summer Stress Tolerance in Tall Fescue (Festuca arundinacea)

Summer stress tolerance (SST) is one of the most important breeding objectives in tall fescue (Festuca arundinacea), an important perennial cool-season grass. However, breeding for better SST is generally complicated by the many environmental factors involved during the growing season. Utilizing the bulked segregant analysis (BSA), we were able to identify one marker related to SST from 100 inter-simple sequence repeat (ISSR) markers and 800 random amplified polymorphic DNA (RAPD) markers, and successfully developed a dominant sequence characterized amplified region (SCAR) marker T_SC856 from the UBC856 sequence. Furthermore, the SCAR marker was tested in different clones of new populations, which were identified under complex summer stress (high temperature and humidity, Pythium blight, and brown patch), and it exhibited relatively high consistency (77%) with the phenotype. We believe that with more markers obtained in the future, better efficiency is likely to be achieved in breeding for improved SST in tall fescue and possibly other species as well. Further studies that analyze the factors relating to the SCAR marker are needed

Reduced circulating interleukin 35 is associated with enhanced peripheral T cell function in primary biliary cholangitis

Interleukin 35 (IL-35) mediates immunosuppression of T cells in autoimmune diseases. T cells play an important role in primary biliary cholangitis (PBC) with incompletely elucidated pathogenesis. Thus, we aimed to investigate the role of IL-35 regulation on T cells in PBC patients. Fifty-one PBC patients and 28 controls were enrolled in this study. Plasma IL-35 level was measured. Purified peripheral CD4+ and CD8+ T cells were stimulated with exogenous IL-35 to investigate their functional phenotypes. IL-35-treated CD8+ T cells were cultured with human intrahepatic biliary epithelial cell line to determine the cytotoxicity of CD8+ T cells from PBC patients. Plasma IL-35 concentration was lower in PBC patients and negatively correlated with alkaline phosphatase. CD4+ T cells from PBC patients exhibited elevated transcription factor expressions and cytokine secretion, whereas CD8+ T cells produced increased cytotoxic molecules and cytokines. In vitro IL-35 stimulation suppressed the production of IL-17 and IL-22 by CD4+ T cells from PBC patients. CD8+ T cells treated with IL-35 mediated reduced target cell death in the direct contact co-culture system in PBC patients. This process was accompanied by reduced production of cytotoxic molecules and cytokines and increased expressions of immune checkpoint receptors in CD8+ T cells. Reduced circulating IL-35 might be insufficient to suppress T cell function, leading to the immune dysregulation in PBC patients

A preliminary evaluation of targeted nanopore sequencing technology for the detection of Mycobacterium tuberculosis in bronchoalveolar lavage fluid specimens

ObjectiveTo evaluate the efficacy of targeted nanopore sequencing technology for the detection of Mycobacterium tuberculosis(M.tb.) in bronchoalveolar lavage fluid(BALF) specimens.MethodsA prospective study was used to select 58 patients with suspected pulmonary tuberculosis(PTB) at Henan Chest Hospital from January to October 2022 for bronchoscopy, and BALF specimens were subjected to acid-fast bacilli(AFB) smear, Mycobacterium tuberculosis MGIT960 liquid culture, Gene Xpert MTB/RIF (Xpert MTB/RIF) and targeted nanopore sequencing (TNS) for the detection of M.tb., comparing the differences in the positive rates of the four methods for the detection of patients with different classifications.ResultsAmong 58 patients with suspected pulmonary tuberculosis, there were 48 patients with a final diagnosis of pulmonary tuberculosis. Using the clinical composite diagnosis as the reference gold standard, the sensitivity of AFB smear were 27.1% (95% CI: 15.3-41.8); for M.tb culture were 39.6% (95% CI: 25.8-54.7); for Xpert MTB/RIF were 56.2% (95% CI: 41.2-70.5); for TNS were 89.6% (95% CI: 77.3-96.5). Using BALF specimens Xpert MTB/RIF and/or M.tb. culture as the reference standard, TNS showed 100% (30/30) sensitivity. The sensitivity of NGS for pulmonary tuberculosis diagnosis was significantly higher than Xpert MTB/RIF, M.tb. culture, and AFB smear. Besides, P values of <0.05 were considered statistically significant.ConclusionUsing a clinical composite reference standard as a reference gold standard, TNS has the highest sensitivity and consistency with clinical diagnosis, and can rapidly and efficiently detect PTB in BALF specimens, which can aid to improve the early diagnosis of suspected tuberculosis patients

Protective Roles of Gadd45 and MDM2 in Blueberry Anthocyanins Mediated DNA Repair of Fragmented and Non-Fragmented DNA Damage in UV-Irradiated HepG2 Cells

Peer reviewe

Protective Roles of Gadd45 and MDM2 in Blueberry Anthocyanins Mediated DNA Repair of Fragmented and Non-Fragmented DNA Damage in UV-Irradiated HepG2 Cells

Peer reviewe

Comparative Genomic Analysis of Latilactobacillus curvatus and L. sakei

In this study, the genomes of 19 Latilactobacillus curvatus and 40 L. sakei strains were comparatively analyzed. Average nucleic acid identity (ANI) and genome-wide colinearity indicated that the genomes of L. curvatus and L. sakei had weak nucleotide sequence homology, allowing them to be used as indicators to distinguish the two species. Pangenomes for these species were constructed, whose core gene functions were annotated. The results showed that the core genomes of L. curvatus and L. sakei were mainly involved in their basic metabolism. Analysis of individual genomes of the strains revealed that 1) both L. curvatus and L. sakei contained a wide range of genes encoding glycoside hydrolases, which are abundant genetic resources for catabolizing and metabolizing dietary fiber such as polysaccharides, lactose utilization, and lignocellulose; 2) antibiotic resistance genes were annotated in the genomes of three strains, which originate from horizontal gene transfer; 3) the unique arginine deiminase pathway of L. sakei, the serine dehydratase and guanine deaminase pathways of L. curvatus, and the glutamate decarboxylase pathway of several strains were identified, revealing that the acid tolerance mechanisms of these two species are different; and 4) genes encoding cold stress proteins were discovered, which endow the two species with good cold processing properties. Moreover, the genomes of some L. sakei strains contained gene clusters related to the biosynthesis of lactocin S and condensin. In conclusion, this study established taxonomic criteria for the two species and information on individual differences between their strains, which will provide a basis for the study of the physiological, biochemical, molecular genetic mechanisms of L. curvatus and L. sakei and their industrial applications

Spatial and Temporal Variations of Potential Evapotranspiration in the Loess Plateau of China During 1960–2017

Potential evapotranspiration (ET0) is an integral component of the hydrological cycle and the global energy balance, and its long-term variation is of much concern in climate change studies. The Loess Plateau is an important area of agricultural civilization and water resources research. This study analyzed the spatial and temporal evolution processes and influential parameters of ET0 at 70 stations in different topographical areas of the Chinese Loess Plateau (CLP). Using the Mann–Kendall trend, Cross wavelet transform, and the ArcGIS platform, the ET0 of each station was quantified using the Penman–Monteith equation, and the effects of climatic factors on ET0 were assessed by analyzing the correlation coefficients and contribution rates of the climatic factors. The results showed that: (1) the overall trend of the ET0 in different terrains of the Loess Plateau is consistent, however, the ET0 values differ; the hill region (HR) has the highest ET0, followed by the valley region (VR), and the mountain region (MR) has the lowest, and ET0 changes differ between seasons. (2) Spatial distribution characteristics of multiyear mean ET0 in the study are as follows: the ET0 values in mountain and hilly areas are decreasing from west to east, and the higher mean annual ET0 value in the VR is mainly concentrated in the eastern CLP. (3) In the past 58 years, the annual mean and the seasonal ET0 of the region showed increasing trends, however, differences in different terrains were obvious. (4) ET0 has significant correlations with El Niño–Southern Oscillation (ENSO), Pacific–North American teleconnection (PNA), and Atlantic Multidecadal Oscillation (AMO). The resonance period of ET0 and ENSO was 3–6 a, mainly in 1976–1985. The mean coherence phase angle was close to 360°, indicating that ET0 lags behind PNA by approximately 2–6 a; ET0 has a very strong positive correlation with AMO. (5) Relative humidity (RH) is the main influencing factor of ET0 change in the Loess Plateau. Temperature (T) variation has the highest contribution rate (42%) to the regional ET0 variation in the entire CLP. We should pay more attention to the variation of evaporation under future climate change, especially temperature change

Heat Prediction of High Energy Physical Data Based on LSTM Recurrent Neural Network

High-energy physics computing is a typical data-intensive calculation. Each year, petabytes of data needs to be analyzed, and data access performance is increasingly demanding. The tiered storage system scheme for building a unified namespace has been widely adopted. Generally, data is stored on storage devices with different performances and different prices according to different access frequency. When the heat of the data changes, the data is then migrated to the appropriate storage tier. At present, heuristic algorithms based on artificial experience are widely used in data heat prediction. Due to the differences in computing models of different users, the accuracy of prediction is low. A method for predicting future access popularity based on file access characteristics with the help of LSTM deep learning algorithm is proposed as the basis for data migration in hierarchical storage. This paper uses the real data of high-energy physics experiment LHAASO as an example for comparative testing. The results show that under the same test conditions, the model has higher prediction accuracy and stronger applicability than existing prediction models