Induction Stage Research Articles

#2581 Cryptococcosis in kidney transplant recipients: a nationwide multicenter retrospective cohort study—CRITERE study

Abstract Background and Aims Cryptococcosis stands as the third most common invasive fungal infection in solid organ transplant recipients. Clinical management and prognosis of the disease remains largely unexplored in the context of kidney transplant recipients. Our main objective was to describe the natural history of cryptococcosis after kidney transplantation from the graft patterns of kidney injury to the long term outcomes. Method We established a nationwide cohort of adult kidney transplant recipients diagnosed with cryptococcosis between 2002 and 2019 in France. All data were gathered starting from the transplantation date and encompassed up to the latest follow-up consultation. All putative causes of AKI as claimed by the attending clinicians were recorded. Cases were matched (ratio 1:6) with kidney transplant recipients from the French prospective multicenter cohort DIVAT with parameters at time of transplant including immunosuppression and known risk factors of cryptococcosis such as age, history of smoke, history of cancer and HIV status. Transplant outcomes (graft failure and patient death) were then compared between matched patients. Results 88 patients were included. Patients were predominantly male (n = 61, 69.3%), with a mean age of 57.0 years [SD ±13.3]. The majority of patients were primary kidney transplants (n = 71, 80.7%) with a median time from ESRD to transplantation of 2.8 years [IQ 1.25-5]. The majority of patients received ATG as induction therapy for their transplant (n = 50, 69.4%)Acute kidney injury (AKI) at diagnosis involved 37.3% of the patients with 13.3% requiring renal replacement therapy. AKI staging was distributed as follows, AKI grade 1 in 12.5% of patients, grade 2 in 13.3% of patients, and grade 3 in 11.5% of patients. During the induction stage, 39% of patients developed amphotericin B-related AKI. AKI resulting from tacrolimus overdose subsequent to tacrolimus/fluconazole interaction occurred in 25.7% of patients during the consolidation stage. Among the patients alive at 3 months following the diagnosis of cryptococcosis, 33.7% of patients exhibited graft dysfunction. Following cryptococcosis, 5 patients displayed T cell rejection and 4 patients developed antibody-mediated rejection. A total of 41 patients with cryptococcosis were matched to 246 controls (ratio 1:6), after a median follow-up of 52.3 [28.1-90.2] months, patients with cryptococcosis exhibited a higher risk of mortality with 35.9% compared with 9.8% (p < 0.001) and graft failure with 33.3% compared with 15.0% (p = 0.0005). Conclusion This study has shown that kidney injury manifests itself through a sequential, multi-step process that initially includes pre-renal impairment, ATN and iatrogenic mechanisms. Amphotericin B-related AKI, followed by tacrolimus overdose represent common causes of AKI.

Comprehensive network modeling approaches unravel dynamic enhancer-promoter interactions across neural differentiation.

Increasing evidence suggests that a substantial proportion of disease-associated mutations occur in enhancers, regions of non-coding DNA essential to gene regulation. Understanding the structures and mechanisms of regulatory programs this variation affects can shed light on the apparatuses of human diseases. We collected epigenetic and gene expression datasets from seven early time points during neural differentiation. Focusing on this model system, we constructed networks of enhancer-promoter interactions, each at an individual stage of neural induction. These networks served as the base for a rich series of analyses, through which we demonstrated their temporal dynamics and enrichment for various disease-associated variants. We applied the Girvan-Newman clustering algorithm to these networks to reveal biologically relevant substructures of regulation. Additionally, we demonstrated methods to validate predicted enhancer-promoter interactions using transcription factor overexpression and massively parallel reporter assays. Our findings suggest a generalizable framework for exploring gene regulatory programs and their dynamics across developmental processes. This includes a comprehensive approach to studying the effects of disease-associated variation on transcriptional networks. The techniques applied to our networks have been published alongside our findings as a computational tool, E-P-INAnalyzer. Our procedure can be utilized across different cellular contexts and disorders.

Calcined alunite-modified alkali-sulphate-activated slag as a novel binder for high-performance cemented paste backfill

The free water content and pore structure significantly affect the mechanical properties of cementitious materials. Because of high water-to-cement ratio and low binder dosage, cemented paste backfill (CPB) is characterized by high free water content and high porosity, which lead to a low strength. In this study, calcine alunite (CA) was used to modify the alkali-sulphate-activated slag (ASAS) to synthesis high-strength CPB with low porosity and low free water content. The results show that at dosage ≤3 %, CA can significantly enhance the strength of ASAS-CPB, and the strengthening effect becomes more prominent as both the dosage and curing time rise. An addition of 3 % CA leads to a 184 % increase in strength at 14 days. However, excessive CA (5 %) reduces 3-day strength by 65 %, but provides the greatest strength at 56 days. As the dosage increases up to 3 %, the promoting impact of CA on early hydration of ASAS-CPB gradually increases, and the duration of induction stage and acceleration stage gradually decreases. Nevertheless, 5 % CA prolongs the duration of induction stage by 921 %. Electrical conductivity and volume water content can be used to assessing the hydration process of ASAS-CPB. Besides, the addition of CA can promote the hydration of slag, resulting a greater formation of C–S–H and ettringite. The hydration products fill the pores and transform the macropores into micropores. Furthermore, mechanisms of CA-induced enhancement in strength is discussed. The results of this study can offer a reference for the synthesis of high performance CPB.

The basic characteristics of paste backfill materials based on highly active mineral admixtures: Part I. Preliminary study on flow, mechanics, hydration and microscopic properties

The cost performance of backfill materials in coal mines has long limited the widespread use of backfill technology. Therefore, developing backfill materials that are both cost-effective and high-performing is a pressing scientific issue that requires immediate attention. This study proposes replacing ordinary Portland cement (OPC) with highly active mineral admixtures, such as mineral powder and limestone powder, to prepare a green, low-cost, and high-performance coal mine backfill material. The use of mineral admixtures can improve the performance of the backfill material while reducing its environmental impact. The proposed approach is cost-effective and environmentally friendly. The study employed experimental tests and microscopic characterization methods, such as the slump test, hydration heat test, uniaxial compressive strength test, and microstructure test, to clarify the synergistic reaction mechanism of highly active mineral admixture-based paste backfill material (HAM-PBM). This clarification is helpful in regulating the macroscopic properties of hardened paste. The results show that: (1) The rheological properties of fresh HAM-PBM slurry are consistent with the Herschel-Bulkley model, and its fluidity meets the technical requirements for coal mine backfilling. The slump of fresh HAM-PBM slurry ranges from 140 to 145 mm, and the yield stress ranges from 3.188 to 12.819 Pa. (2) Highly active mineral admixtures significantly affect the early hydration process of HAM-PBM. The parameters of the hydration characteristics during the initial dissolution stage, acceleration stage, deceleration stage, and stabilization stage gradually decrease, while the duration of the induction stage gradually increases. This confirms that the early hydration activity of mineral powder (MP) and limestone powder (LF) is lower than that of OPC. (3) The uniaxial compressive strength (UCS) of the MP series exhibits an "anti-hook" change pattern, while the UCS of the LF series shows a decreasing change pattern. The microstructure, including the number of hydration products, microscopic density, and other characteristics, follows the same change pattern as the UCS. (4) Based on the synergistic reaction mechanism of hardened slurry, it is recommended that MP replace a maximum of 10 % of OPC and LF replace a maximum of 5 % of OPC. The need for composite replacement of OPC with MP and LF is clarified, resulting in a cost-effective coal mine backfill material. This study has contributed to the national "double carbon" policy and the low-cost backfilling application of coal mines.

Exploring miR-21 as a key regulator in two distinct approaches of bone marrow stromal cells differentiation into Schwann-like cells.

The differentiation of bone marrow stromal cells (BMSCs) into Schwann-like cells (SCLCs) has the potential to promote the structural and functional restoration of injured axons. However, the optimal induction protocol and its underlying mechanisms remain unclear. This study aimed to compare the effectiveness of different induction protocols in promoting the differentiation of rat BMSCs into SCLCs and to explore their potential mechanisms. BMSCs were induced using two distinct methods: a composite factor induction approach (Protocol-1) and a conditioned culture medium induction approach (Protocol-2). The expression of Schwann cells (SCs) marker proteins and neurotrophic factors (NTFs) in the differentiated cells was assessed. Cell proliferation and apoptosis were also measured. During induction, changes in miR-21 and Sprouty RTK signaling antagonist 2 (SPRY2) mRNA were analyzed. Following the transfection of BMSCs with miR-21 agomir or miR-21 antagomir, induction was carried out using both protocols, and the expression of SPRY2, ERK1/2, and SCs marker proteins was examined. The results revealed that NTFs expression was higher in Protocol-1, whereas SCs marker proteins expression did not significantly differ between the two groups. Compared to Protocol-1, Protocol-2 exhibited enhanced cell proliferation and fewer apoptotic and necrotic cells. Both protocols showed a negative correlation between miR-21 and SPRY2 expression throughout the induction stages. After induction, the miR-21 agomir group exhibited reduced SPRY2 expression, increased ERK1/2 expression, and significantly elevated expression of SCs marker proteins. This study demonstrates that Protocol-1 yields higher NTFs expression, whereas Protocol-2 results in stronger SCLCs proliferation. Upregulating miR-21 suppresses SPRY2 expression, activates the ERK1/2 signaling pathway, and promotes BMSC differentiation into SCLCs.

Comparative transcriptome profiling reveals that light coordinates auxin to inhibit adventitious root formation in grapevine

Grapevine (Vitis sp.) is one of the most important economic fruit crops all over the world, and the formation of adventitious roots (ARs) is crucial for the vegetative reproduction of grapes. However, studies on the regulatory mechanisms of this process are currently lacking. In this study, we applied an efficient and convenient leave-petiole (LP) system for studying ARs, revealing a significant inhibition of root primordia formation under continuous-light treatment. The results showed that isolated ARs of grapevine were induced and originated from ray cells near the vascular cambium, with the process categorized into induction, initiation, and extension stages. LP samples under light and dark conditions were used for transcriptome sequencing and endogenous hormone measurements at three critical time points of AR formation. A total of 37 155 transcripts were obtained, and 7041 genes showed significantly different expression levels in the petiole. An integrated analysis, including Gene Ontology (GO) enrichment analysis, weighted gene co-expression network analysis (WGCNA), and hormonal content determination, showed that several genes (ARF4, LAX1, PIN1, SUS2, APX1, TPXL1, CHS3, etc.) associated with hormone signals, sugar synthesis and transport, reactive oxygen species (ROS) scavenging, cell wall biogenesis, flavonoid biosynthesis, microtubule remodeling, and some transcription factors (HY5, COP1, ERF2, MYB15, etc) played vital roles in light-induced AR formation. A hypothetical model was initially constructed, which illustrated the centrality of auxin in HY5-dependent AR formation and the complex crosstalk among various factors. The results of this study provided abundant genetic resources and a novel perspective for understanding the molecular mechanisms of AR formation in grapevine.

Disaggregation and fibrillation during sol-gel transition of alginate hydrogels

The rheological and morphological characteristics of Ca-crosslinked alginate hydrogels with two different M/G ratios, α-L-guluronate (G)-rich and β-D-mannuronate (M)-rich, each with one alginic acid concentration, were investigated. It was found that the stiffness and elasticity of alginate hydrogels are derived from the thickness and density of the fibril network structures. In aqueous alginate solution, ball-like aggregates of alginates are present. Time-resolved small-angle X-ray scattering and time-domain nuclear magnetic resonance measurements suggest that the disaggregation of alginate aggregates and loose fibrillation occur in the early stage of the sol-gel transition. After these induction stage, direct gelation is finally caused by the formation of the egg-box junction. G-rich alginate hydrogel has a higher stiffness and a thicker and denser fibril network structure than M-rich alginate hydrogel. The former also exhibits faster and more significant changes in physical properties during the sol-gel transition.

Recycling multisource industrial waste residues as green binder for cemented ultrafine tailings backfill: Hydration kinetics, mechanical properties, and solidification mechanism

Incorporating mine tailings with industrial waste residues (IWRs) is progressively used to manufacture filling materials, which provides an eco-friendly disposal method for IWRs and alleviate their impact on the environment. In this study, steel slag (5, 10, 15, 20 wt%), fly ash (5, 10 wt%) and blast furnace slag (52, 57, 65, 74 wt%) as the precursor, carbide slag (16, 18, 20 wt%) and desulfurization gypsum (0, 2, 3 wt%) as the activator were reutilized to synthesize the IWRs-based binder for cemented ultrafine tailings backfill (CUFTB). Experimental tests were conducted to determine the setting time, hydration heat release, mechanical behavior, and microstructure characteristics. The hydration kinetics and solidification mechanism were expounded correspondingly. Results elucidate that the initial and final setting time of CUFTB are respectively 12.08–13.92 h and 16.92–20 h <24 h much longer than the transport time in pipeline and meets the operational requirements of mine site. Hydration process of CUFTB includes initial dissolution, induction, acceleration, deceleration, and decline stages. As the hydration proceeds, the initial stage of CUFTB is regulated by nucleation and crystal growth (NG), and gradually shifted to interactions at phase boundaries (I) or diffusion (D). The high content of carbide slag (20 wt%) promotes the hydration reaction, resulting in the increase of heat release. IWRs-based binder hydrates to form calcite, ettringite, hydroaluminite, and gehlenite. Especially, the gel products are mainly C-A-S-H, C-(Al,Na)-S-H, C-(Al,K)-S-H with Ca/Si ratio <1, demonstrating a fundamental distinction from Portland cement. Under the bonding and padding of hydration products, tailings particles are solidified well. After curing of 28 days, CUFTB exhibit high UCS values exceeding 2.0 MPa, presenting an evident improvement of 26.70% ∼ 39.20% compared to those made of Portland cement. These findings are referential for preparation of cemented paste backfill by reutilizing IWRs in mine filling field.

Features of hemodynamics in elderly patients with coronary heart disease using a combination of propofol, ketamine and fentanyl for induction anesthesia

The aim – to analyze hemodynamic changes in elderly patients with coronary heart disease during coronary artery bypass grafting surgery without cardiopulmonary bypass at the stage of induction into anesthesia using propofol, ketamine and fentanyl.Materials and methods. The results of the examination of 20 patients with coronary artery disease over 60 years of age who underwent coronary artery bypass grafting without the use of artificial circulation are presented. The effect of anesthesia on hemodynamics was assessed by mean arterial pressure (MAP), heart rate (HR), cardiac index (CI), and systemic vascular resistance index (SVRI). The following drugs were used for induction: Propofol 1.5 mg/kg, Ketamine 0.5 mg/kg, Fentanyl 2.0 μg/kg, Pipecuronium bromide 0.1 mg/kg. Anesthesia maintenance: Propofol 4 mg/kg/hour, Fentanyl 2.0 μg/kg/hour.Results and discussion. At the time of admission to the surgery room, hemodynamic parameters reflected that the general condition of patients was stable. After the administration of the combination of drugs for induction propofol + fentanyl + ketamine in the dosage according to the study design, it was found that MAP decreased by 18.71 % (p&lt;0,0001), heart rate decreased by 4.72 % (p = 0,2199), CI decreased by 14.37 % (p = 0,0035), and SVRI decreased by 3.93 % (p = 0,41). After insertion of the intubation tube, it was determined that MAP was 92.34±7.26 % of the baseline values (p = 0,0001), heart rate was 100.81±9.77 % of the baseline data (p = 0,848), CI was determined at 110.38±12.37 % of baseline value (p = 0,0769), and SVRI was 83.57±10.7 % of the baseline, respectively. In 25 minutes after successful tracheal intubation, the parameters were re-registered again: MAP = 86.47±6.07 % of the baseline (p &lt; 0,0001); heart rate = 100.08±5.03 % of the baseline value (p = 0,9711); CI = 108.29±9.95 % of baseline data (p = 0,1248); SVRI = 77.86±9.83 % of the baseline (p = 0,0001).Conclusions. Since hypotension is a frequent side effect of traditional drugs for induction of anesthesia, such as propofol and fentanyl, the addition of ketamine to this combination helps to stabilize hemodynamic parameters and prevents undesirable hemodynamic fluctuations.

N-acetylcysteine Protects Against Myocardial Ischemia–Reperfusion Injury Through Anti-ferroptosis in Type 1 Diabetic Mice

The hearts of subjects with diabetes are vulnerable to ischemia–reperfusion injury (IRI). In contrast, experimentally rodent hearts have been shown to be more resistant to IRI at the very early stages of diabetes induction than the heart of the non-diabetic control mice, and the mechanism is largely unclear. Ferroptosis has recently been shown to play an important role in myocardial IRI including that in diabetes, while the specific mechanisms are still unclear. Non-diabetic control (NC) and streptozotocin-induced diabetic (DM) mice were treated with the antioxidant N-acetylcysteine (NAC) in drinking water for 4 week starting at 1 week after diabetes induction. Mice were subjected to myocardial IRI induced by occluding the coronary artery for 30 min followed by 2 h of reperfusion, subsequently at 1, 2, and 5 week of diabetes induction. The post-ischemic myocardial infarct size in the DM mice was smaller than that in NC mice at 1 week of diabetes but greater than that in the NC mice at 2 and 5 week of diabetes, which were associated with a significant increase of ferroptosis at 2 and 5 week but a significant reduction of ferroptosis at 1 week of diabetes. NAC significantly attenuated post-ischemic ferroptosis as well as oxidative stress and reduced infarct size at 2 and 5 week of diabetes. Application of erastin, a ferroptosis inducer, reversed the cardioprotective effects of NAC. It is concluded that increased oxidative stress and ferroptosis are the major factors attributable to the increased vulnerability to myocardial IRI in diabetes and that attenuation of ferroptosis represents a major mechanism whereby NAC confers cardioprotection against myocardial IRI in diabetes.

The design of an Obstetric Telephone Triage Guideline (OTTG): a mixed method study

BackgroundClarifying the dimensions and characteristics of obstetric telephone triage is important in improving the quality of services in the health system because researchers can evaluate the effectiveness of treatment, care and diagnostic measures in the form of obstetric telephone triage by developing a guideline. Therefore, this study aimed to design an Obstetric Telephone Triage Guideline (OTTG) using a mixed-method study.MethodsThe present study was carried out using an exploratory sequential mixed method study in two qualitative and quantitative phases. An inductive-deductive approach was also used to determine the concept of obstetric telephone triage. In this respect, a qualitative study and a literature review were used in the inductive and deductive stages, respectively. Moreover, the validity of the developed guideline was confirmed based on experts’ opinions and results of the AGREE II tool.ResultsThe guideline included the items for evaluating the severity of obstetric symptoms at five levels including “critical”, “urgent”, “less urgent”, “no urgent”, and “recommendations”. The validity of the guideline was approved at 96%, 95%, 97%, 95%, 93%, and 100% for six dimensions of AGREE II including scope and purpose, stakeholder involvement, the rigor of development, clarity of presentation, applicability, and editorial independence, respectively.ConclusionThe OTTG is a clinically comprehensive, easy-to-use, practical, and valid tool. This guideline is a standardized tool for evaluating the severity of symptoms and determining the urgency for obstetrics triage services. By using this integrated and uniform guideline, personal biases can be avoided, leading to improved performance and ensuring that patients are not overlooked. Additionally, the use of OTTG promotes independent decision-making and reduces errors in triage decision-making.

Sinonasal diffuse large B-cell lymphoma: own clinical observation and literature review

Diffuse large B-cell lymphoma is the most common immunomorphological variant of lymphoma in adults. Extranodal lesions are observed in a third of patients at the disease onset. The organs most often involved are the gastrointestinal tract, testicles, bones, thyroid gland, and skin. Primary involvement of the nasal cavity and paranasal sinuses occur extremely rarely and cause diagnostic and therapeutic difficulties.The article demonstrates a rare clinical case of newly diagnosed diffuse large B-cell lymphoma with sinonasal tract involvement. It took 6 months to verify the final diagnosis. At the moment, the induction stage of treatment for diffuse large B-cell lymphoma continues, the achieved complete metabolic response is maintained.

Experimental study on cement-based materials for grouting/replacement of broken rock mass in coal mine

In deep high-pressure mines, the surrounding rock of soft broken roadways often faces issues such as poor anchorability, support difficulties, high maintenance costs, and the inadequacy of traditional materials for site requirements. To tackle these challenges, this study focuses on grouting reinforcement in coal mine sites and prefabricated bearing arch form (grouting or replacement) under specific conditions, aiming to meet the material requirements. Through an orthogonal test, various properties of cement-based materials were investigated, including fluidity, initial setting time, water separation rate, early (≤1d) compressive strength, fracture strength, and later (28d as a reference) strength. The objective was to develop a cost-effective cement-based material with high strength and toughness, suitable for underground coal mines. The experimental results demonstrate that the self-developed cement-based materials exhibit rapid setting, excellent fluidity, high early strength, and strong durability compared to traditional alternatives. Analyses of the comprehensive properties reveal that the influencing factors, in order of significance, include the mass ratio of cement and sand, water cement ratio, rubber content, and bonding mortar master material content. By employing extreme treatment and efficiency coefficient methods, the comprehensive performance of materials under different ratios was evaluated. Consistent findings were obtained, indicating that the optimal mass ratio of cement-based materials is 500:500:20:15:10:5:8:10 for cement, sand, fly ash, silica fume, mineral powder, steel slag powder, bonding mortar master material, and rubber, respectively. Additionally, the best comprehensive performance was achieved with a water cement ratio of 0.5, which confirms the results of the orthogonal test. Macroscopic failure characteristics and SEM microscopic analysis further validate the superior performance of the materials under the aforementioned ratio. The reaction process of the material can be roughly divided into four stages: initial hydrolysis stage, induction stage, acceleration stage, decay and stability stage, and finally the formation of stone body. A comparison with ordinary materials highlights the advantages of the developed cement-based materials, and suggestions are proposed for the development of field-applicable materials.

THE COST OF ACUTE LYMPHOBLASTIC LEUKEMIA TREATMENT IN CHILDREN USING COMBINED CHEMOTHERAPY AND IMMUNOTHERAPY: COMPARISON OF ESTIMATED AND REAL VALUES

Acute lymphoblastic leukemia is the most common pediatric cancer. The effectiveness of modern therapy protocols makes it possible to cure over 80% of children, and one of the advanced treatment methods is the use of monoclonal antibodies. Using the example of adult patients in the United States, the economic effectiveness of this innovative treatment strategy was proven, while no similar studies have been conducted in children with primary acute lymphoblastic leukemia. The aim of the study was to determine the real cost of therapy for children and adolescents with acute lymphoblastic leukemia according to the protocol using combined chemotherapy and immunotherapy at the stages of induction and consolidation. Materials and methods. The study included data from 54 patients (the study group) who received therapy using this method, taking into account indicators of direct medical and non-medical costs. The control group included 54 patient models corresponding to the initial estimated parameters when planning the protocol (body weight 30 kg, body surface area 1 m2). Research results. The median of real cost for providing medical care to one patient in the study group was 22% lower than the estimated cost. In its structure, the values of the median costs for inpatient stay, for instrumental and laboratory studies, as well as for drug therapy turned out to be lower. Since younger patients were included in the study group, the cost of drug therapy was recalculated to the calculated patient model, which equalized the median cost of drug therapy for the study and control groups. In a situation with equal characteristics of patients, as well as with the same cost of a day of inpatient stay, the median real cost of providing medical care to one patient in the study group corresponded to the estimated values. Conclusions. The median cost of medical care for patients with acute lymphoblastic leukemia according to the protocol using combined chemotherapy and immunotherapy in the study group was 22% lower than the estimated values. The actual practice of using the protocol for an average patient model with an equal cost of inpatient stay fully corresponded to the estimated values of the protocol.

Type IV Collagen Promotes Adipogenic Differentiation of Adipose Stem Cells.

Type IV collagen is a major component of the extracellular matrix in adipose tissue. It is secreted during the lipogenic differentiation of mesenchymal stem cells, but its direct impact and mechanism on the differentiation of adipose-derived stem cells (ASCs) into lipids are unclear. In this study, ASCs were obtained from human liposuction samples and cultured. Lipogenic induction of ASCs was achieved using lipogenic induction medium. Immunofluorescence analysis revealed differential expression of type IV collagen during the early and late stages of adipogenic induction, displaying a distinct morphological encapsulation of ASCs. Silencing of type IV collagen using siRNA resulted in a significant decrease in adipogenic capacity, as indicated by reduced lipid droplet formation and downregulation of adipogenic-related gene transcription. Conversely, supplementation of the culture medium with synthetic type IV collagen demonstrated enhanced adipogenic induction efficiency, accompanied by upregulation of YAP/TAZ protein expression and its downstream target gene transcription. Furthermore, inhibition of the YAP/TAZ pathway using the inhibitor Blebbistatin attenuated the functionality of type IV collagen, leading to decreased lipid droplet formation and downregulation of adipocyte maturation-related gene expression. These findings highlight the crucial role of type IV collagen in promoting adipogenic differentiation of ASCs and suggest its involvement in the YAP/TAZ-mediated Hippo pathway.No Level Assigned This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Identification and functional analysis of the LEAFY gene in longan flower induction

BackgroundFlowering at the right time is a very important factor affecting the stable annual yield of longan. However, a lack of knowledge of the regulatory mechanism and key genes of longan flowering restricts healthy development of the longan industry. Therefore, identifying relevant genes and analysing their regulatory mechanism are essential for scientific research and longan industry development.ResultsDlLFY (Dimocarpus longan LEAFY) contains a 1167 bp open reading frame and encodes 388 amino acids. The amino acid sequence has a typical LFY/FLO family domain. DlLFY was expressed in all tissues tested, except for the leaf, pericarp, and pulp, with the highest expression occurring in flower buds. Expression of DlLFY was significantly upregulated at the early flower induction stage in “SX” (“Shixia”). The results of subcellular localization and transactivation analysis showed that DlLFY is a typical transcription factor acting as a transcriptional activator. Moreover, overexpression of DlLFY in Arabidopsis promoted early flowering and restrained growth, resulting in reduced plant height and rosette leaf number and area in transgenic plants. DNA affinity purification sequencing (DAP-Seq) analysis showed that 13 flower-related genes corresponding to five homologous genes of Arabidopsis may have binding sites and be putative target genes. Among these five flower-related genes, only AtTFL1 (terminal flower 1) was strongly inhibited in transgenic lines.ConclusionTaken together, these results indicate that DlLFY plays a pivotal role in controlling longan flowering, possibly by interacting with TFL1.

Enhancing lipid production in Chlorella under successive stresses of periodic micro-current and salinity: Performance and contribution

Two-stage cultivation of Chlorella with periodic micro-current coupled with NaCl stress was investigated to analyze the performance and contribution of increased lipid production. In the growth stage, periodic micro-current of 3 mA resulted in an escalation of 290.06 mg/L of Chlorella’s biomass by 61.25 % in comparison to the control. During the induction stage, under a NaCl concentration of 8 g/L, the highest lipid content and yield attained were 42.11 % and 199.32 mg/L, revealing an increment of 53.13 % and 39.11 % respectively when compared to the control. Additionally, the polyunsaturated fatty acids content (PUFA) decreased from 30.11 % to 23.09 %, while the saturated fatty acids content (SFA) increased from 31.86 % to 42.39 %. Chlorella demonstrated adaptability to the alterations in the cultivation environment by an initial upsurge and later a reduction in SOD and CAT enzyme activities, and concurrently facilitated lipid synthesis by similar changes in the activities of ACCase and DGAT. The above cultivation enabled a synergistic effect of swift growth and efficient lipid accumulation in Chlorella, providing groundwork for large-scale biofuel production.

Enhancing in vitro regeneration via somatic embryogenesis and Fusarium wilt resistance of Egyptian cucumber (Cucumis sativus L.) cultivars

BackgroundSomatic embryogenesis offers a reliable method for cucumber (Cucumis sativus L.) regeneration and genetic enhancement against Fusarium wilt. This study aimed to establish a tailored somatic embryogenesis system for Egyptian cultivars, fostering genetic improvements and Fusarium wilt-resistance lines. ResultsEmploying the Optimal Arbitrary Design (OAD) approach, we optimized the induction medium, initiating prolific embryogenic calli (53.3 %) at 1 mg/L 2,4-D. The cotyledonary leaf (CL) was the preferred explant, showing 60 % embryogenic callus development. Bieth Alpha exhibited higher responsiveness, generating ∼ 18 somatic embryos per explant compared to Prince's ∼ 10. Somatic embryogenesis system validation used quantitative RT-PCR, showing Cucumis sativus splicing factor 3B subunit (CUS1) and an embryogenesis marker gene expression exclusively within embryogenic calli and mainly during embryogenesis initiation. Evaluating fungal toxin filtrate concentrations for selecting embryogenic calli, the S2 selection (25 % filtrate, four subculture cycles) was chosen for somatic embryo development. To gauge the ramifications of selection at the genetic stratum, an in-depth analysis was executed. A cluster analysis grounded in ISSR banding patterns revealed a distinct separation between in vivo-cultivated plants of the two cultivars and regenerated plants devoid of pathogen filtrate treatment or those regenerated post-filtrate treatment. This segregation distinctly underscores the discernible genetic impact of the selection process. ConclusionsThe highest embryogenic capacity (53.3%) was achieved in this study by optimizing the induction stage, which demonstrated the optimal concentrations of BA and 2,4-D for induced proembryonic masses. Moreover, consistent gene expression throughout both stages of embryogenesis suggests that our system unequivocally follows the somatic embryogenesis pathway.

Role of neurotrophic growth factors in vincristine polyneuropathy in children with acute lymphoblastic leukemia

Vincristine-induced peripheral neuropathy (VIPN) is one of the frequent toxic complications in the treatment of acute lymphoblastic leukemia in children. The pathogenesis of peripheral nerve damage is not fully understood; however, recent studies have demonstrated the involvement of neurotrophic factors. The purpose of the study: to evaluate the level of plasma neurotrophic growth factors in children with acute lymphoblastic leukemia (ALL) and determine their association with the VIPN formation. Materials and methods: 131 newly diagnosed ALL patients aged 3 to 17 years receiving chemotherapy according to the ALL–MB 2015 protocol participated in a single-center prospective study. Depending on the development of VIPN, the patients were divided into two groups: the study group (n=106) — children with VIPN and the comparison group (n=25) — children without VIPN. The plasma level of neurotrophic growth factors (NGF-β and BDNF) was determined using multiparametric immunofluorescence analysis.Results: during the follow-up period, 80.9 % of the children (n=106) developed VIPN against the background of chemotherapy. In most cases, neurotoxic disorder manifested at the induction stage of treatment — in 84.9 % (n=90) of the patients. The clinical phenotype of VIPN was characterized by a combination of neurological disorders in 67.9 % (n=72) of the patients, with the predominance of sensory and motor symptoms. The comparative analysis of plasma neurotrophic growth factors in the groups of children with ALL depending on the VIPN formation showed that in the patients with VIPN, a statistically significant increase in brain-derived neurotrophic factor (BDNF) was noted at the consolidation stage of chemotherapy (study 1–284.3 (97.4÷628.3) pg/mL; study 2–281.7 (178.9÷679.2) pg/mL; study 3–980.2 (454.3÷2,005.9) pg/mL; p1-2=0.424 and p1-3=0.009). However, in the children without VIPN, an increase in this growth factor was observed during the induction phase (study 1–370.5 (95.4÷463.8) pg/mL; study 2–683.0 (362.4÷1,486.3) pg/mL; study 3–674.6 (394.8÷2,584.0) pg/mL; p1-2=0.043 and p1-3=0.021). In addition, in the study group patients with the early debut of VIPN, the level of nerve growth factor-β (NGF-β) before the administration of chemotherapy was significantly lower in contrast to the patients with its development in later terms (22.7 (10.9÷22.7) pg/mL and 24.7 (22.7÷91.5) pg/mL, respectively; p=0.045). When assessing the clinical value of this indicator, the diagnostic sensitivity was 88 %, specificity — 71 %, and the integral index characterizing the accuracy of the test was 0.81. Conclusion. The increase in plasma growth factor (BDNF) in earlier terms in children without VIPN probably reflects the mechanisms aimed at preventing the realization of VIPN. The established low concentration of plasma NGF-β and optimal diagnostic characteristics of the factor in children with early VIPN onset allow considering it as a prognostic biomarker.

Study of the Formation of Hydrates with NaCl, Methanol Additive, and Quartz Sand Particles

During deepwater drilling, testing, production, or hydrate mining, the circulating medium in the wellbore may contain solid particles, such as rock chips and sand, in addition to drilling fluids, gas, and water. In the high-pressure, low-temperature conditions of deep water, gas intrusion can easily combine with free water in the drilling fluid to form hydrates, increasing the drilling risk. Therefore, understanding the formation patterns of hydrates in drilling fluids is of significant importance for the prevention and control of hydrates. This study utilized a small-scale high-pressure reactor to investigate the impact of the stirring rate, NaCl, and methanol additives, as well as the sand content on the hydrate formation process and gas consumption. The results indicate that the hydrate formation process can be divided into an induction stage, a rapid formation stage, and a slow formation stage. The induction stage and rapid formation stage durations are significantly reduced under stirring conditions. In NaCl and methanol solutions, hydrate formation is inhibited, with the induction stage duration increasing with higher concentrations of NaCl and methanol. There was no apparent rapid formation stage observed. The final gas consumption decreases substantially with increasing concentrations of NaCl and methanol, reaching no significant hydrate formation at a 20% concentration. The sand content has a significant impact on the slow formation stage, with the final gas consumption increasing within a certain range (in this work, at a sand content of 20%), and being notably higher than in the pure water system under the same conditions.