The expression of miR-21, HSP90a and gGASP-1 in serum of patients with lung cancer and their correlation with pathological subtypes.

Background: To investigate the expression of miR-21, heat shock protein-90a (HSP90a) and G protein-coupled receptorrelated sorting protein 1(GASP-1) in the serum of lung cancer patients and their correlation with pathological subtypes. Methods: Eighty patients with lung cancer were included in the lung cancer group from May 2020 to May 2022, and 40 volunteers who underwent physical examination were randomly included in the control group according to the group ratio of 2:1. This ratio balances the need for a sufficiently large experimental group to detect significant effects with the practicality of recruiting a manageable control group. To ensure the validity of our findings, we meticulously calculated the sample size to achieve adequate statistical power, thus enabling us to draw reliable conclusions. Serum miR-21, HSP90a and GASP-1 levels of patients in the two groups were detected. We quantitatively assessed the serum levels of miR-21, HSP90a, and GASP1 in lung cancer patients and healthy volunteers. We employed enzyme-linked immunosorbent assay (ELISA) for HSP90a and GASP-1, and reverse transcription-polymerase chain reaction (RT-PCR) for miR-21, ensuring precise quantification. To explore the correlation between it and pathological subtypes, TNM stage and lymph node metastasis of lung cancer patients. TNM stands for Tumor, Node, and Metastasis. This system is widely used for staging cancer and describes the size and extent of the primary tumor (T), the absence or presence of cancer in nearby lymph nodes (N), and whether the cancer has spread to other parts of the body (M). Results: The serum levels of miR-21, HSP90a and GASP1 in lung cancer group were higher than those in control group (P < 0.05). ROC curve analysis showed that serum miR-21, HSP90a and GASP-1 levels had certain value in the diagnosis of lung cancer, and their AUC values were 0.901, 0.874 and 0.865, respectively (P < 0.05). There was no difference in the relative expression level of serum miR-21 between squamous cell carcinoma group and adenocarcinoma group (P>0.05), but the levels of HSP90a and GASP-1 in adenocarcinoma group were higher than those in squamous cell carcinoma group (P < 0.05). There was no difference in the levels of serum miR-21, HSP90a and GASP-1 between stage I and stage II groups (P>0.05). The levels of serum miR-21, HSP90a and GASP-1 in stage III and stage IV groups were higher than those in stage I and stage II groups, and those in stage IV were higher than those in stage III group (P < 0.05). The serum levels of miR-21, HSP90a and GASP-1 in patients with metastasis were higher than those in patients without metastasis (P < 0.05). Conclusions: Our study concludes that there is a notable association between elevated serum levels of miR-21, HSP90a, and GASP-1 and lung cancer. However, it is crucial to acknowledge that these findings are preliminary and further statistical analysis is needed to strengthen these associations. Future studies with comprehensive statistical evaluation will be vital to validate these potential biomarkers for lung cancer diagnosis and prognosis.

Ketoconazole induces reversible antifungal drug tolerance mediated by trisomy of chromosome R in Candida albicans.

Background: The emergence of tolerance to antifungal agents in Candida albicans complicates the treatment of fungal infections. Understanding the mechanisms underlying this tolerance is crucial for developing effective therapeutic strategies. Objective: This study aims to elucidate the genetic and molecular basis of ketoconazole tolerance in C. albicans, focusing on the roles of chromosomal aneuploidy, Hsp90, and calcineurin. Methods: The wild-type C. albicans strain SC5314 was exposed to increasing concentrations of ketoconazole (0.015-32 µg/mL) to select for tolerant adaptors. Disk diffusion and spot assays were used to assess tolerance. Whole-genome sequencing identified chromosomal changes in the adaptors. The roles of Hsp90 and calcineurin in maintaining and developing ketoconazole tolerance were investigated using specific inhibitors and knockout strains. Results: Adaptors exhibited tolerance to ketoconazole concentrations up to 16 µg/mL, a significant increase from the parent strain's inhibition at 0.015 µg/mL. All tolerant adaptors showed amplification of chromosome R, with 29 adaptors having trisomy and one having tetrasomy. This aneuploidy was unstable, reverting to euploidy and losing tolerance in drug-free conditions. Both Hsp90 and calcineurin were essential for maintaining and developing ketoconazole tolerance. Inhibition of these proteins resulted in loss of tolerance. The efflux gene CDR1 was not required for the development of tolerance. Chromosome R trisomy and tetrasomy induce cross-tolerance to other azole antifungal agents, including clotrimazole and miconazole, but not to other antifungal classes, such as echinocandins and pyrimidines, exemplified by caspofungin and 5-flucytosine. Conclusion: Ketoconazole tolerance in C. albicans is mediated by chromosomal aneuploidy, specifically chromosome R amplification, and requires Hsp90 and calcineurin. These findings highlight potential targets for therapeutic intervention to combat antifungal tolerance and improve treatment outcomes.

ClassIIb histone deacetylase participates in perioperative neurocognitive disorders in elderly mice via HSP90/GR signaling pathway.

OBJECTIVE: Multiple factors contribute to the development of perioperative neurocognitive disorders (PND). This study was designed to investigate whether Histone Deacetylase 6 (HDAC6) was involved in the formation of postoperative cognitive dysfunction in elderly mice by regulating the degree of acetylation of heat shock protein (HSP90) and related protein functions and quantities. METHODS: C57BL/6 J male mice were randomly divided into six groups: control naive (group Control), anesthesia (group Anesthesia), splenectomy surgery (group Surgery), splenectomy surgery plus dissolvent (group Vehicles), splenectomy surgery plus the inhibitor ACY-1215 (group Ricolinostat), and splenectomy surgery plus the inhibitor RU-486(group Mifepristone). After the mice were trained for Morris Water Maze (MWM) test for five days, anesthesia and operational surgery were carried out the following day. Cognitive function was assessed on the 1st, 3rd and 7th days post-surgery. The hippocampi were harvested on days 1, 3, and 7 post-surgeries for Western blots and ELISA assays. RESULTS: Mice with the splenectomy surgery displayed the activation of the hypothalamic-pituitary-adrenal axis (HPA-axis), marked an increase in adrenocorticotropic hormone (ACTH), glucocorticoid, mineralocorticoid at the molecular level and impaired spatial memory in the MWM test. The hippocampus of surgical groups showed a decrease in acetylated HSP90, a rise in glucocorticoid receptor (GR)-HSP90 association, and an increase in GR phosphorylation and translocation. HDAC6 was increased after the surgical treated. Using two specific inhibitors, HDAC6 inhibitor Ricolinostat (ACY-1215) and GR inhibitor Mifepristone (RU-486), can partially mitigate the effects caused by surgical operation. CONCLUSIONS: Abdominal surgery may impair hippocampal spatial memory, possibly through the HDAC6-triggered increase in the function of HSP90, consequently strengthening the negative role of steroids in cognitive function. Targeting HDAC6- HSP90/GR signaling may provide a potential avenue for the treatment of the impairment of cognitive function after surgery.

The Investigation of Hsp90 C-terminal inhibitors containing Amide bioisosteres.

Heat Shock Protein 90 (Hsp90) is responsible for the proper folding and maturation of ~400 client protein substrates, many of which are directly associated with the ten hallmarks of cancer. Hsp90 is a great target for cancer therapy including melanoma, since Hsp90 inhibition can disrupt multiple oncogenic pathways simultaneously. In this study, we report the synthesis and anti-proliferative activity manifested by a series of Hsp90 C-terminal inhibitors against mutant BRAF and wild-type BRAF melanoma cells. Furthermore, we explored structure-activity relationships (SAR) for the amide moiety of 6 (B1), a novel Hsp90 C-terminal inhibitor via introduction of amide bioisosteres. Compound 6 displayed an IC50 of 1.01 µM, 0.782 µM, 0.607 µM and 1.413 µM against SKMel173, SKMel103, SKMel19 and A375 cells, respectively.

Modulating the HSP90 control over NFκB/NLRP3/Caspase-1 axis is a new therapeutic target in the management of liver fibrosis: Insights into the role of TAS-116 (Pimitespib).

Aberrant activation of the NLRP3 inflammasome is recognized to induce a chronic inflammatory response in the liver, ultimately leading to hepatic fibrosis. HSP90 is suggested to regulate NLRP3 activation and its downstream signaling. This study is the first to explore the potential therapeutic role of pimitespib in mitigating liver fibrosis in rats. The results of the study revealed that pimitespib effectively suppressed hepatic inflammation and fibrogenesis by modulating HSP90's control over the NFκB/NLRP3/caspase-1 axis. In vitro experiments demonstrated that pimitespib reduced LDH levels and increased hepatocyte survival, whereas in vivo, it prolonged the survival of rats with hepatic fibrosis. Additionally, pimitespib exhibited improvements in the function and microscopic characteristics of rat livers. Pimitespib effectively inhibited NFκB, which serves as the priming signal for NLRP3 activation. Pimitespib's inhibitory effect on NLRP3, identified as an HSP90 client protein, plays a central role in the observed anti-fibrotic effect. The simultaneous inhibition of both priming and activation signals of NLRP3 by pimitespib led to a reduction in caspase-1 activity and subsequent suppression of the N-terminal fragment of gasdermin D, ultimately constraining hepatocyte pyroptotic cell death. These diverse effects were associated with a decrease in the transcription of inflammatory mediators IL-1ß, IL-18, and TNF-α, as well as the fibrogenic mediators TGF-ß, TIMP-1, PDGF-BB, and Col1a1. Moreover, pimitespib induced the expression of HSP70, which could further contribute to the repression of fibrosis development. In summary, our findings provide an evolutionary perspective on managing liver fibrosis, positioning pimitespib as a promising candidate for anti-inflammatory and antifibrotic therapy.

HSP90 multi-functionality in cancer.

The 90-kDa heat shock proteins (HSP90s) are molecular chaperones essential for folding, unfolding, degradation and activity of a wide range of client proteins. HSP90s and their cognate co-chaperones are subject to various post-translational modifications, functional consequences of which are not fully understood in cancer. Intracellular and extracellular HSP90 family members (HSP90α, HSP90ß, GRP94 and TRAP1) promote cancer by sustaining various hallmarks of cancer, including cell death resistance, replicative immortality, tumor immunity, angiogenesis, invasion and metastasis. Given the importance of HSP90 in tumor progression, various inhibitors and HSP90-based vaccines were developed for the treatment of cancer. Further understanding of HSP90 functions in cancer may provide new opportunities and novel therapeutic strategies for the treatment of cancer.

AUY922 improves sensitivity to sunitinib in clear cell renal cell carcinoma based on network pharmacology and in vitro experiments.

Clear Cell Renal Cell Carcinoma (ccRCC), the most prevalent form of renal cell carcinoma (RCC), poses a significant threat to human health due to its rising morbidity and mortality rates. Sunitinib, a pivotal targeted drug for the treatment of ccRCC, presents a significant challenge due to the high susceptibility of ccRCC to resistance. HSP90 inhibitor AUY922 has demonstrated anti-tumor activity in a range of cancer types. However, its efficacy in combination with sunitinib for ccRCC treatment has not been evaluated. In this study, we employed bioinformatics, network pharmacology, and in vitro assays to verify that AUY922 inhibits cell viability, proliferation, and migration of ccRCC cell lines 786-O and ACHN, with IC50s of 91.86 µM for 786-O and 115.5 µM for ACHN. The effect of AUY922 enhancing the inhibitory effect of sunitinib on ccRCC was further confirmed. The CCK-8 assay demonstrated that the IC50 of sunitinib was reduced from 15.10 µM to 11.91 µM for 786-O and from 17.65 µM to 13.66 µM for ACHN, after the combined application of AUY922. The EdU assay and wound healing assay indicated that AUY922 augmented the inhibitory impact of sunitinib on the proliferation and migration of ccRCC cells. Western blot and RT-PCR analyses demonstrated that AUY922 increased the sensitivity of ccRCC cells to sunitinib by targeting the HIF-1α/VEGFA/VEGFR pathway. Our study represents the first investigation into the role and mechanism of AUY922 in enhancing the sensitivity of ccRCC to sunitinib. In conclusion, the findings indicate the potential for AUY922 to enhance the therapeutic efficacy of sunitinib and overcome sunitinib resistance in ccRCC.

Leveraging virucidal potential of an anti-microbial coating agent to mitigate fomite transmission of respiratory viruses.

In the wake of the COVID-19 pandemic, respiratory tract infections have emerged as a significant global threat, yet their impact on public health was previously underappreciated. This study investigated the antiviral efficacy of the nano-coating agent BARRIER90, composed of silicon-quaternary ammonium compound and a naturally derived biopolymer, against three distinct respiratory viruses: Influenza A (H1N1), Adenovirus Type 1, and Enterovirus-Coxsackie B1. BARRIER90 exhibited robust and sustained virucidal activity, persisting up to 90 days post-coating, against the enveloped virus, Influenza A, with significant reduction in viral plaques. Contrastingly, its efficacy against non-enveloped viruses revealed transient activity against Enterovirus-Coxsackie B1, with almost no antiviral activity observed against Adenovirus Type 1. These findings indicate the potential of antimicrobial coatings in mitigating viral transmission through contaminated surfaces (fomites), which harbour pathogenic viruses for longer periods. Antimicrobial coatings may facilitate infection control in various settings, including healthcare facilities and shared workspaces.

Evaluation of sensitivity and specificity of the slit-light method in the diagnosis of cyclotorsion.

PURPOSE: The purpose of this study was to evaluate the amount of sensitivity and specificity of the slit-light (SL) method in the diagnosis of ocular cyclotorsion. MATERIALS AND METHODS: One hundred and twenty eyes of 60 individuals (10-60 years old), with mean visual acuity of 0.08 ± 0.14 LogMAR, were divided into two groups (normal and torsion groups). Individuals without ocular motility disorder were selected as normal and patients with extraocular motility disorders and oblique muscle dysfunctions as the torsion group. The sensitivity and specificity of SL in the diagnosis of ocular torsion were measured by masked investigators and compared to fundus photography (FP). Inter- and intraobserver variability of these techniques was also determined. RESULTS: The amounts of sensitivity and specificity of SL, measured by the first examiner, were 60% and 92% for intorsion and 50% and 96% for extorsion assessment, respectively. These amounts were 53% and 95% for intorsion, and 54% and 97% for extorsion by the second examiner. The contingency coefficient between the two examiners was 68.6% for SL. This amount was 61% between FP and SL for the first examiner and 63% for the second. The contingency coefficient for the repeatability of SL was 72.2% for the first examiner and 75.7% for the second. This amount was 71.2% between the two examiners. CONCLUSION: SL can be considered a useful method for the diagnosis of cyclotorsion.

Construction of intelligent response gene vector based on MOF/Fe3O4/AuNRs for tumor-targeted gene delivery.

Metal-organic frameworks (MOFs) have the potential to efficiently carry cargo due to their excellent porosity and high surface area. Nevertheless, conventional MOFs and their derivatives exhibit low efficiency in transporting nucleic acids and other small molecules, as well as having poor colloidal stability. In this study, a ZIF-90 loaded with iron oxide nanoparticles and Au nanorods was prepared, and then surface-functionalized with polyethyleneimine (PEI) to create a multifunctional nanocomposite (AFZP25k) with pH, photothermal, and magnetic responsiveness. AFZP25k can condense plasmid DNA to form AFZP25k/DNA complexes, with a maximum binding efficiency of 92.85 %. DNA release assay showed significant light and pH responsiveness, with over 80 % cumulative release after 6 h of incubation. When an external magnetic field is applied, the cellular uptake efficiency in HeLa cells reached 81.51 %, with low cytotoxicity and specific distribution. In vitro transfection experiments demonstrated a gene transfection efficiency of 44.77 % in HeLa cells. Following near-infrared irradiation, the uptake efficiency and transfection efficiency of AFZP25k in HeLa cells increased by 21.3 % and 13.59 % respectively. The findings indicate the potential of AFZP25k as an efficient and targeted gene delivery vector in cancer gene therapy.

Discovery of Cell Number-Interstitial Fluid Volume (CIF) Ratio Reveals Secretory Autophagy Pathway to Supply eHsp90α for Wound Healing.

Cell secretion repairs tissue damage and restores homeostasis throughout adult life. The extracellular heat shock protein-90alpha (eHsp90α) has been reported as an exosome cargo and a potential driver of wound healing. However, neither the mechanism of secretion nor the genetic evidence for eHsp90α in wound healing has been substantiated. Herein, we show that tissue injury causes massive deposition of eHsp90α in tissues and secretion of eHsp90α by cells. Sequential centrifugations of conditioned medium from relevant cell lines revealed the relative distributions of eHsp90α in microvesicle, exosome and trypsin-sensitive supernatant fractions to be approximately <2%, <4% and >95%, respectively. Establishing the cell-number-to-interstitial-fluid-volume (CIF) ratio for the microenvironment of human tissues as 1 × 109 cells: 1 mL interstitial fluid enabled us to predict the corresponding tissue concentrations of eHsp90α in these fractions as 3.74 µg/mL, 5.61 µg/mL and 178 µg/mL. Remarkably, the 178 µg/mL eHsp90α matches the previously reported 100-300 µg/mL of recombinant eHsp90α whose topical application promotes maximum wound healing in animal models. More importantly, we demonstrate that two parallel secretory autophagy-regulating gene families, the autophagy-regulating (AR) genes and the Golgi reassembly-stacking protein (GRASP) genes work together to mediate the secretion of the physiological concentration of eHsp90α to promote wound healing. Thus, utilization of the CIF ratio-based extrapolation method may enable investigators to rapidly predict biomarker targets from cell-conditioned-medium data.

Changes of circulating tumor cells expressing CD90 and EpCAM in early-phase of atezolizumab and bevacizumab for hepatocellular carcinoma.

Circulating tumor cells (CTCs) are noninvasive biomarkers that can indicate the therapeutic response and prognosis. The study aimed to investigate the cellular characteristics of CTCs focusing on monitoring during atezolizumab and bevacizumab (Atezo-Bev) therapy in patients with hepatocellular carcinoma (HCC). Peripheral blood samples were collected from 10 healthy controls and 40 patients with HCC. CTCs enriched using RosetteSep™ Human CD45 depletion cocktail were analyzed by multiparametric flow cytometry. CTC isolation was based on PanCK(+)CD45(-) cells, and CTCs exhibiting markers CD90, CD133, EpCAM, or vimentin. The total number of CTCs and the number of CTCs expressing CD90, CD133, EpCAM, and vimentin were correlated with the BCLC stage of HCC. The change in total CTC count accurately reflected the initial response to Atezo-Bev therapy. The numbers and mean fluorescence intensity of the CTC subsets expressing CD90 and EpCAM molecules decreased in patients with partial response/stable disease, and increased in patients with progressive disease and were markedly correlated with overall survival. CD90(+) and EpCAM(+) CTCs may be candidate biomarkers for the early prediction of the treatment response and the overall survival of patients with HCC receiving Atezo-Bev therapy.

On tricyclic graphs with maximum atom-bond sum-connectivity index.

The sum-connectivity, Randic, and atom-bond connectivity indices have a prominent place among those topological indices that depend on the graph's vertex degrees. The ABS (atom-bond sum-connectivity) index is a variant of all the aforementioned three indices, which was recently put forward. Let T ( n ) be the class of all connected tricyclic graphs of order n. Recently, the problem of determining graphs from T ( n ) having the least possible value of the ABS index was solved in (Zuo et al., 2024 [39]) for the case when the maximum degree of the considered graphs does not exceed 4. The present paper addresses the problem of finding graphs from T ( n ) having the largest possible value of the ABS index for n ≥ 5 .

Genome-wide identification of HSP90 gene family in Rosa chinensis and its response to salt and drought stresses.

Heat shock protein 90 (HSP90) is important for many organisms, including plants. Based on the whole genome information, the gene number, gene structure, evolutionary relationship, protein structure, and active site of the HSP90 gene family in Rosa chinensis and Rubus idaeus were determined, and the expression of the HSP90 gene under salt, and drought stresses in two rose varieties Wangxifeng and Sweet Avalanche were analyzed. Six and eight HSP90 genes were identified from R. chinensis and Ru. idaeus, respectively. Phylogenetic analysis revealed that the analyzed genes were divided into two Groups and four subgroups (Classes 1a, 1b, 2a, and 2b). Although members within the same classes displayed highly similar gene structures, while the gene structures and conserved domains of Group 1 (Class 1a and 1b) and the Group 2 (Class 2a and 2b) are different. Tandem and segmental duplication genes were found in Ru. idaeus, but not in R. chinensis, perhaps explaining the difference in HSP90 gene quantity in the two analyzed species. Analysis of cis-acting elements revealed abundant abiotic stress, photolight-response, and hormone-response elements in R. chinensis HSP90s. qRT-PCR analysis suggested that RcHSP90-1-1, RcHSP90-5-1 and RcHSP90-6-1 in Sweet Avalanche and Wangxifeng varieties played important regulatory roles under salt and drought stress. The analysis of protein structure and active sites indicate that the potential different roles of RcHSP90-1-1, RcHSP90-5-1, and RcHSP90-6-1 in salt and drought stresses may come from the differences of corresponding protein structures and activation sites. These data will provide information for the breeding of rose varieties with high stress resistance. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-024-04052-0.

Characteristics and outcomes of children ≤ 10 kg receiving continuous kidney replacement therapy: a WE-ROCK study.

BACKGROUND: Continuous kidney replacement therapy (CKRT) is often used for acute kidney injury (AKI) or fluid overload (FO) in children ≤ 10 kg. Intensive care unit (ICU) mortality in children ≤ 10 kg reported by the prospective pediatric CRRT (ppCRRT, 2001-2003) registry was 57%. We aimed to evaluate characteristics associated with ICU mortality using a contemporary registry. METHODS: The Worldwide Exploration of Renal Replacement Outcomes Collaborative in Kidney Disease (WE-ROCK) registry is a retrospective, multinational, observational study of children and young adults aged 0-25 years receiving CKRT (2015-2021) for AKI or FO. This analysis included patients ≤ 10 kg at hospital admission. PRIMARY AND SECONDARY OUTCOMES: ICU mortality and major adverse kidney events at 90 days (MAKE-90) defined as death, persistent kidney dysfunction, or dialysis within 90 days, respectively. RESULTS: A total of 210 patients were included (median age 0.53 years (IQR, 0.1, 0.9)). ICU mortality was 46.5%. MAKE-90 occurred in 150/207 (72%). CKRT was initiated at a median 3 days (IQR 1, 9) after ICU admission and lasted a median 6 days (IQR 3, 16). On multivariable analysis, pediatric logistic organ dysfunction score (PELOD-2) at CKRT initiation was associated with increased odds of ICU mortality (aOR 2.64, 95% CI 1.68-4.16), and increased odds of MAKE-90 (aOR 2.2, 95% CI 1.31-3.69). Absence of comorbidity was associated with lower MAKE-90 (aOR 0.29, 95%CI 0.13-0.65). CONCLUSIONS: We report on a contemporary cohort of children ≤ 10 kg treated with CKRT for acute kidney injury and/or fluid overload. ICU mortality is decreased compared to ppCRRT. The extended risk of death and morbidity at 90 days highlights the importance of close follow-up.

Extracellular heat shock protein 90 alpha (eHsp90α)'s role in cancer progression and the development of therapeutic strategies.

Heat shock protein 90 alpha (Hsp90α) is an abundantly expressed and evolutionarily conserved molecular chaperone. Hsp90α is the inducible Hsp90 isoform, and its expression and secretion extracellularly (eHsp90α) can be triggered in response to a variety of cellular stresses to protect/activate client proteins and to facilitate cellular adjustment to the stress. As a result, cancers often have high expression levels of intracellular and extracellular (plasma) Hsp90α, allowing them to support their oncogenesis and progression. In fact, (e)Hsp90α has been implicated in regulating processes such as cell signaling transduction, DNA repair, promotion of the Epithelial-to-Mesenchymal Transition (EMT), promotion of angiogenesis, immune response, and cell migration. Hsp90α levels have been correlated with cancer progression and severity in several cancers, indicating that it may be a useful biomarker or drug-target for cancer. To date, the development of intracellular Hsp90α-targeted therapies include standard N-terminal ATP-competitive inhibitors and allosteric regulators that bind to Hsp90α's middle or C-terminal domain. On-target toxicities and dosing complications as a result of Hsp90α inhibition has driven the development of eHsp90α-targeted therapies. Examples include anti-Hsp90α monoclonal antibodies and cell-impermeable Hsp90α small molecule inhibitors. This review aims to discuss the many roles Hsp90α plays in cancer progression with a focus on the current development of Hsp90α-targeted therapies.

Exogenous application of selenium on sunflower (Helianthus annuus L.) to enhance drought stress tolerance by morpho-physiological and biochemical adaptations.

Drought stress poses a significant obstacle to agricultural productivity, particularly in the case of oilseed crops such as sunflower (Helianthus annuus L.). Selenium (Se) is a fundamental micronutrient that has been recognized for its ability to enhance plant resilience in the face of various environmental stresses. The FH-770 sunflower variety was cultivated in pots subjected to three stress levels (100% FC, 75% FC, and 50% FC) and four Se application rates (0 ppm, 30 ppm, 60 ppm, and 90 ppm). This research aimed to investigate the effect of exogenously applied Se on morpho-physiological and biochemical attributes of sunflower to improve the drought tolerance. Foliar Se application significantly lowered H2O2 (hydrogen peroxide; ROS) (20.89%) accumulation that markedly improved glycine betaine (GB) (74.46%) and total soluble protein (Pro) (68.63%), improved the accumulation of ascorbic acid (AA) (25.51%), total phenolics (TP) (39.34%), flavonoids (Flv) (73.16%), and anthocyanin (Ant) (83.73%), and improved the activity of antioxidant system superoxide dismutase (SOD) (157.63%), peroxidase (POD) (100.20%), and catalase (CAT) (49.87%), which ultimately improved sunflower growth by 36.65% during drought stress. Supplemental Se significantly increased shoot Se content (93.86%) and improved calcium (Ca2+), potassium (K+), and sodium (Na+) ions in roots by 36.16%, 42.68%, and 63.40%, respectively. Selenium supplements at lower concentrations (60 and 90 ppm) promoted the growth, development, and biochemical attributes of sunflowers in controlled and water-deficient circumstances. However, selenium treatment improved photosynthetic efficiency, plant growth, enzymatic activities, osmoregulation, biochemical characteristics, and nutrient balance. The mechanisms and molecular processes through which Se induces these modifications need further investigation to be properly identified.

Type II mode of JAK2 inhibition and destabilization are potential therapeutic approaches against the ruxolitinib resistance driven myeloproliferative neoplasms.

Background: Ruxolitinib has been approved by the US FDA for the treatment of myeloproliferative neoplasms such as polycythemia vera and primary myelofibrosis. Ruxolitinib will remain a main stay in the treatment of MPN patients due to its effective therapeutic benefits. However, there have been instances of ruxolitinib resistance in MPN patients. As JAK2 is a direct target of ruxolitinib, we generated ruxolitinib-resistant clones to find out the mechanism of resistance. Methods: Cell-based screening strategy was used to detect the ruxolitinib-resistant mutations in JAK2. The Sanger sequencing method was used to detect the point mutations in JAK2. Mutations were re-introduced using the site-directed mutagenesis method and stably expressed in Ba/F3 cells. Drug sensitivities against the JAK2 inhibitors were measured using an MTS-based assay. JAK2 and STAT5 activation levels and total proteins were measured using immunoblotting. Computational docking studies were performed using the Glide module of Schrodinger Maestro software. Results: In this study, we have recovered seven residues in the kinase domain of JAK2 that affect ruxolitinib sensitivity. All these mutations confer cross-resistance across the panel of JAK2 kinase inhibitors except JAK2-L983F. JAK2-L983F reduces the sensitivity towards ruxolitinib. However, it is sensitive towards fedratinib indicating that our screen identifies the drug-specific resistance profiles. All the ruxolitinib-resistant JAK2 variants displayed sensitivity towards type II JAK2 inhibitor CHZ-868. In this study, we also found that JAK1-L1010F (homologous JAK2-L983F) is highly resistant towards ruxolitinib suggesting the possibility of JAK1 escape mutations in JAK2-driven MPNs and JAK1 mutated ALL. Finally, our study also shows that HSP90 inhibitors are potent against ruxolitinib-resistant variants through the JAK2 degradation and provides the rationale for clinical evaluation of potent HSP90 inhibitors in genetic resistance driven by JAK2 inhibitors. Conclusion: Our study identifies JAK1 and JAK2 resistance variants against the type I JAK2 inhibitors ruxolitinib, fedratinib, and lestaurtinib. The sensitivity of these resistant variants towards the type II JAK2 inhibitor CHZ-868 indicates that this mode of type II JAK2 inhibition is a potential therapeutic approach against ruxolitinib refractory leukemia. This also proposes the development of potent and specific type II JAK2 inhibitors using ruxolitinib-resistance variants as a prototype.

Predictors of emergency abdominal surgery for patients aged 90 years or older: A retrospective study.

Background: With the aging of the population, more and more patients ≥90 years old are undergoing surgery. We retrospectively examined factors affecting morbidity and in-hospital mortality among patients ≥90 years old who underwent emergency abdominal operations. Materials and methods: Forty-six cases of emergency abdominal surgery for patients ≥90 years old who underwent surgery at our hospital between 2011 and 2022 were included in this study. Factors affecting morbidity and in-hospital mortality were analyzed statistically. Physiological and operative severity score for the enumeration of mortality and morbidity (POSSUM)-predicted morbidity and Portsmouth-POSSUM (P-POSSUM)-predicted mortality were calculated. Results: Postoperative complications occurred in 30 patients (65.2 %) and 5 patients (10.8 %) died in the hospital. Factors affecting morbidity included American Society of Anesthesiologists physical status score, operative time and blood loss, and operative severity score. Multivariate analysis identified male sex, operative severity score, and length of hospital stay as factors affecting morbidity. Eastern Cooperative Oncology Group performance status and physiological score were identified as factors influencing mortality in hospital, and only physiological score was identified in the multivariate analysis. Area under the receiver operating characteristic (ROC) curve for POSSUM-predicted morbidity was 0.796 and area under the ROC curve for P-POSSUM-predicted mortality was 0.805, both of which were moderately accurate. Conclusion: Risk of emergency abdominal surgery in patients ≥90 years old may be predictable to some extent, and we are able to provide convincing explanations to patients and families based on these data.

Long-term outcomes and associated factors among intensive care unit survivors in a low-income country: a multicenter prospective cohort study.

OBJECTIVE: Post-ICU survivors face higher mortality and often require costly rehabilitation or palliative care, such as occupational therapy, physiotherapy and hospice. However, there is a lack of data quantifying the demand for these services, particularly in developing countries like Uganda. Therefore, this prospective cohort study aimed to investigate the 90-day mortality rate, functional status, and mortality risk factors among 121 ICU patients discharged from three tertiary hospital ICUs in Uganda by tracking their vital and physical functional status for three months with follow-ups on days 30, 60, and 90, and identifying risk factors through Cox regression. RESULTS: The study revealed that 18 out of 121 ICU patients (14.88%, 95% CI: 9.52-22.51%) died within 90 days post-discharge, while 36.36% achieved normal physical functional status. Factors associated with higher 90-day mortality included raised intracranial pressure (HR 1.92, 95% CI: 1.76-2.79, p = 0.04), acute kidney injury (HR 4.13, 95% CI: 2.16-7.89, p < 0.01), and renal replacement therapy (HR 3.34, 95% CI: 2.21-5.06, p < 0.01). The high mortality rate and the fact that nearly two-thirds of patients did not attain normal functional status 90 days post discharge underscores the need for enhanced post-ICU rehabilitation services.