B1 Levels Research Articles

Anticancer Study of a Novel Pan-HDAC Inhibitor MPT0G236 in Colorectal Cancer Cells.

Colorectal cancer (CRC) is one of the most commonly diagnosed malignancies and a leading cause of cancer worldwide. Histone deacetylases (HDACs), which regulate cell proliferation and survival, are associated with the development and progression of cancer. Moreover, HDAC inhibitors are promising therapeutic targets, with five HDAC inhibitors approved for cancer treatment to date. However, their safety profile necessitates the exploration of well-tolerated HDAC inhibitors that can be used in cancer therapeutic strategies. In this study, the pan-HDAC inhibitor MPT0G236 reduced the viability and inhibited the proliferation of human colorectal cancer cells, and normal human umbilical vein endothelial cells (HUVECs) showed reduced sensitivity. These findings indicated that MPT0G236 specifically targeted malignant tumor cells. Notably, MPT0G236 significantly inhibited the activities of HDAC1, HDAC2, and HDAC3, Class I HDACs, as well as HDAC6, a Class IIb HDAC, at low nanomolar concentrations. Additionally, it promoted the accumulation of acetyl-α-tubulin and acetyl-histone H3 in HCT-116 and HT-29 cells in a concentration-dependent manner. Furthermore, MPT0G236 treatment induced G2/M cell cycle arrest in CRC cells by initially regulating the levels of cell-cycle-related proteins, such as p-MPM2; specifically reducing p-cdc2 (Y15), cyclin B1, and cdc25C levels; and subsequently inducing apoptosis through the caspase-dependent pathways and PARP activation. Our findings demonstrate that MPT0G236 exhibits significant anticancer activity in human colorectal cancer cells.

Development and validation of a QuEChERS-LC-MS/MS method for determination of multiple mycotoxins in maize and sorghum from Botswana.

Climatic conditions such as drought, high temperatures, and pre-harvest rainfalls promote the occurrence of mycotoxins in grains. Contamination of staple food sources such as maize and sorghum means that many populations are at risk of being poisoned by mycotoxins. Hence the need for sensitive methods for their simultaneous analysis. Herein, a quick, easy, cheap, effective, rugged, and safe liquid chromatography tandem mass spectrometry (QuEChERS-LC-MS/MS) method for the simultaneous determination of ten mycotoxins in maize and sorghum is presented. The QuEChERS extraction procedure was optimized to maximize extraction recovery and minimize matrix effects while using relatively small quantities of organic solvents and acids. This method was validated according to Commission Implementing Regulation (EU) 2021/808, Commission Regulation (EC) No 1881/2006, and Regulation (EC) no. 401/2006. The developed method met the specified requirements. Recoveries of 80.77% to 109.83% and CVs below 15% were obtained. The correlation coefficient values (R2) were all above 0.98, and low limits of quantification ranging from 0.53 to 89.28 µg/Kg were recorded. The method was applied to 10 maize and 10 sorghum samples collected from markets in Botswana. Half of the samples had detectable mycotoxins, Aflatoxins, Fumonisins, T2-toxin, HT2-toxin, and Zearalenone. Two maize samples had levels of aflatoxin B1 above the maximum permitted level (2.55, 4.07 µg/Kg). These findings point to the necessity of more stringent monitoring of mycotoxins, particularly AFB1 in maize, as well as the value of regular assessment using LC-MS/MS.

Effect of the extra-nuclear cation on the cytotoxicity and mechanism of action of pyridine-2,6-dicarboxylate Ga(III) complexes

Two Ga(III) complexes (C1) and (C2) were prepared by the one-pot reaction of pyridine-2,6-dicarboxylic acid and aminopyridine derivatives with gallium(III) nitrate octahydrate. The compounds were characterized by single-crystal X-ray diffraction. The distorted octahedral geometry was confirmed by crystallographic data for both complexes. The study of the in vitro cytotoxicity of the compounds showed that the presence of different extra-nuclear cations can affect the cytotoxicity of the same anionic complexes. The most significant antiproliferative activity was observed for C1 (IC50 = 0.69 μM, MAE = 73.96%) and C2 (IC50 = 3.78 μM, MAE = 60.35%) (where MAE represents the maximal antiproliferative effect) against A431 cell line. The mechanistic study evidenced the same pathway for the death of A431 cells treated with the complexes, although the results for C2 were obtained at approximately five times the concentration of C1. According to the study, both complexes induced cell cycle arrest in G2/M phase in A431 cells by upregulating the levels of p21, p27, p-cdc25C, and p-cdc2 and downregulating the levels of cdc25C, cdc2, and cyclin B1. In addition, apoptosis via a caspase-dependent mitochondrial pathway was confirmed by a decrease in Bcl-2 family proteins and an increase in the expression of procaspase-9 and 3. Also, the complexes induced autophagic cell death by activating the RAGE /PI3KC3/Beclin 1 pathway in A431 cells. Data availabilityCCDC 874052 and 874055 contain the supplementary crystallographic data for C1 and C2, respectively. These data can be obtained free of charge via http://www.ccdc.cam.ac.uk/services/structures?pid=ccdc:874052,874055&sid=CCDCManual, or from the Cambridge Crystallographic Data Centre, 12 Union Road, Cambridge CB2 1EZ, UK; fax: (+44) 1223–336–033; or e-mail: deposit@ccdc.cam.ac.uk.

Anti-inflammatory effect of ApoE23 on Salmonella typhimurium-induced sepsis in mice.

Two independent experiments were performed with three groups each (sepsis control, sepsis, and sepsis with apoE23 treatment) to investigate the anti-inflammatory effect of apolipoprotein 23 (apoE23) in a mouse model of sepsis induced by S. typhimurium. Survival rates; plasma level variations in tumor necrosis factor (TNF)-α, interleukin (IL)-6, and lipopolysaccharide (LPS); S. typhimurium colony-forming units in the spleen tissue; and mRNA and protein expression levels of low-density lipoprotein receptor (LDLR), LDLR-related protein (LRP), syndecan-1, and scavenger receptor B1 were evaluated in the livers of mice from the three groups. Results found that the survival rate of septic mice treated with apoE23 was 100% within 48 h, while it was only 40% in septic mice without apoE23 treatment (P < 0.001). The plasma LPS, TNF-α, and IL-6 levels and the S. typhimurium load in mice in the apoE23-treated group were significantly lower than those in septic mice (P < 0.05). Moreover, apoE23 restored the downregulated expression of LDLR and LRP in the liver tissue of septic mice. So apoE23 exhibits an anti-inflammatory effect in the mouse model of S. typhimurium-induced sepsis. Further studies are required to understand the mechanisms underlying the anti-inflammatory effects of apoE23.

Encapsulated Zataria multiflora's essential oil inhibited the growth of Aspergillus flavus and reduced aflatoxins levels in contaminated pistachio nut

The essential oil of Zataria multiflora Boiss. (ZEO) was prepared using a British Pharmacopeia-type hydrodistillation apparatus. The ZEO was encapsulated with a natural polymer, chitosan (CS) by oil-in-water emulsification, and ionic cross-linking techniques. The shape and size range of the encapsulated particles were evaluated by Atomic-Force Microscopy (AFM) and Dynamic Light Scattering (DLS) and FT-IR spectroscopy. The AFM analysis of the ZEO encapsulated nano-particles (CS-ZEO) showed spherical shape particles with inner cavities. The mean size of CS-ZEO particles was 293 ± 17 nm according to DLS analysis. The chemical constituent of the oil was analyzed by GC-MS and thymol acetate (32.4%), carvacrol acetate (19.0%), γ-terpinene (18.5%), and ρ-cymene (14.5%) were reported as its major constituents. The non-encapsulated ZEO in the synthesis process was determined to calculate the encapsulation efficiency and loading capacity of the prepared CS-ZEO which were 87.4% and 6%; respectively. In vitro antifungal activity of ZEO and CS-ZEO in growth media against Aspergillus flavus was evaluated with the decrease in fungal mycelial mass. On the other hand, the HPLC results showed the reduction of aflatoxins B1 levels by 95% and 92% at concentrations of 0.8 mg/mL of ZEO and 6.7 mg/mL of CS-ZEO, respectively. The above-mentioned effective concentrations of the ZEO and CS-ZEO, were applied to pistachio samples, which reduced 90.5% and 99.0%, of the aflatoxin B1 levels, respectively compared to non-treated samples. Therefore, we suggest here, the CS-ZEO as a possible natural fungicide with an effective mechanism of lowering aflatoxins production in the contaminated pistachio.

Determination of Multimycotoxin in Cereal-Based Products Sold in Open-Air Markets.

In this study, a total of 140 cereal-based foods sold in temporary open-air markets were analyzed by LC-MS/MS for aflatoxin B1, B2, G1, G2, ochratoxin (OTA), zearalenone (ZEN), deoxynivalenol (DON), fumonisin B1, fumonisin B2, citrinin (CIT), HT-2, and T-2 toxins. Breakfast cereals (n:27), cornmeal (n:41), extruded maize (n:32), and oatmeal (n:40) purchased from these alternative shopping areas created to meet the food needs of low-income people in the suburbs formed the sample set of the study. These foods, which are sold in areas that are out of legal control and greatly affected by external environmental conditions, are more open to health risks. Mycotoxins, chemicals of a biological origin, are some of the most important of these risks. In terms of public health, it is important to investigate the presence of mycotoxins in foods, which can cause acute and chronic diseases such as immunosuppression, genotoxic, estrogenic, teratogenic effect, cancer, and liver and kidney dysfunctions. Grain-based foods are often contaminated with a large number of mycotoxins, but legal regulations have not been prepared that consider the health risks associated with the co-existence of mycotoxins. Many of the studies have focused on the presence of a single mycotoxin and the risks it poses. As a result, aflatoxin B1 levels in 28.57% of the samples and total aflatoxin (B1 + B2 + G1 + G2) levels in 26.43% of the samples were determined to exceed the limits defined in the "Turkish Food Codex Contaminants Regulation". Citrinin could not be detected in any of the samples. The rate of mycotoxin occurrences above the limit of detection (LOD) in grain-based food samples ranged from 22.86% to 99.29%. Total aflatoxin (TAF) + Total Fumonisin (FUM) were found in 83.57% of the samples; TAF + FUM + OTA in 82.14%; TAF + FUM + OTA + T-2 in 44.29%; TAF + FUM + OTA + DON + HT-2, TAF + FUM + OTA + DON + T-2, and TAF + FUM + OTA + DON + ZEN in 22.86% of the samples.

Inhibition of Cell Proliferation and Cell Death by Apigetrin through Death Receptor-Mediated Pathway in Hepatocellular Cancer Cells.

Epidemiologic research recommends using flavonoids in the diet due to their overall health benefits. Apigetrin (Apigenin 7-O-glucoside) is a glycoside phytonutrient found in fruits and vegetables and known for different biological activities such as antioxidant and anti-inflammatory properties. Hepatocellular cancer (HCC) is a major health concern because of its adverse prognosis and side effects of chemotherapeutic agents. In the present study, we determine the impact of apigetrin on HepG2 cells and its cell death mechanism. Apigetrin reduced HepG2 cell proliferation with morphological changes and floating cells in treated cells. Colony formation and wound healing assays showed a reduced cell number in treatment groups. Further, we checked for the cell cycle through flow cytometry to understand the cell death mechanism. Apigetrin induced G2/M phase arrest in HepG2 cells by regulating Cyclin B1 and CDK1 protein levels in HepG2 cells. Annexin V and propidium iodide (PI) staining was performed to confirm the apoptotic cell population in treated groups. At the higher concentration, apigetrin showed a late apoptotic population in HepG2 cells. Chromatin condensation was also found in the treatment groups. Western blot analysis showed an increased expression of extrinsic apoptotic proteins such as FasL, Cleaved caspase 8, Cleaved caspase 3, and cleavage of PARP. In comparison, intrinsic apoptotic pathway markers showed no changes in Bax, Bcl-xL, and Cleaved caspase 9. Altogether, these findings strongly indicate that apigetrin causes cell death in HepG2 cells through the extrinsic apoptotic pathway, and that the intrinsic/mitochondrial pathway is not involved.

Analysis and Health Risk Evaluation of Aflatoxin B1 Levels in Groundnut (Arachis hypogea ) and Maize (Zea mays ) Samples from Wukari, Nigeria

Aflatoxin B1 (AFB1) is a secondary fungal metabolite which is considered a public health concern worldwide due to its genotoxicity and carcinogenicity. In this study, we evaluated the levels and potential health effects of AFB1 in Groundnut and Maize samples from Wukari, Nigeria. Ten samples (each) of maize and groundnuts were collected randomly from local markets and analysed for AFB1. Our findings revealed the presence of AFB1 in all the samples investigated with concentrations ranging between 7.79 – 14.08 μg/kg in groundnuts and 1.48 -15.50 μg/kg in maize samples. Overall, 90% of the analysed samples contained aflatoxin B1 above the allowed limit of 5 μg/kg. The assessment of chronic exposure and probalistic health risk from consumption of AFB1 in the investigated food samples was done via chronic daily intake (CDI) and margin of exposure (MOE) evaluations. Obtained CDI values for groundnut ranged between 14.96 – 28.74 ng·kg−1 BW day−1 for children and 3.20 – 6.15 ng·kg−1 BW day−1 for adults while for maize the values were in the range of 5.38 – 56.42 ng·kg−1 BW day−1 for children and 1.15 – 12.09 ng·kg−1 BW day−1 for adults. MOE values ranged between 3.01 – 31.60 for children and 14.06 – 147.83 for adults. The obtained MOE values are by far lower that the recommended ≥10,000 value thus indicating high carcinogenic risks to consumers of the food items. There is need to create more awareness and interventions on aflatoxin contamination of food sources in Nigeria.

The correlation between the severity of cerebral microbleeds and serum HMGB1 levels and cognitive impairment in patients with cerebral small vessel disease.

The study investigated the correlation and predictive value between the severity of cerebral microbleeds (CMBs) and the level of serum High Mobility Group Protein B1 (HMGB1) and the occurrence of cognitive impairment in patients with cerebral small vessel disease (CSVD). A total of 139 patients with CSVD admitted to the Department of Neurology of the First Affiliated Hospital of Xinxiang Medical University from December 2020 to December 2022 were selected as study subjects. The Montreal Cognitive Assessment (MoCA) scale was used to assess the cognitive function and was divided into the cognitive impairment group and the cognitive normal group. Magnetic Resonance Imaging (MRI) and Susceptibility Weighted Imaging (SWI) were used to screen and assess the severity of CMBs. Serum HMGB1 levels of CSVD patients were measured by enzyme linked immunosorbent assay (ELISA). Multivariable logistic regression analysis was used to explore risk factors for cognitive impairment and CMBs. Pearson correlation analysis was used to investigate the correlation between HMGB1 and cognitive function. Receiver Operating Characteristics (ROC) curves were used to assess the predictive value of HMGB1 for the occurrence of cognitive impairment in patients with CMBs. High Mobility Group Protein B1, uric acid (UA), glycosylated hemoglobin (HbA1c), CMBs, lacunar cerebral infarction (LI), years of education, and history of hypertension were risk factors for cognitive impairment (P < 0.05); HMGB1 was significantly and negatively associated with total MoCA score, visuospatial/executive ability, and delayed recall ability (P < 0.05). HMGB1 was significantly and positively correlated with the number of CMBs (P < 0.05). The area under the ROC curve for HMGB1 predicting cognitive impairment in patients with CMBs was 0.807 (P < 0.001). Serum HMGB1 levels are associated with the development of cognitive impairment in CSVD patients, and serum HMGB1 levels have a high predictive value for the development of cognitive impairment in CSVD patients with combined CMBs, which can be used for early clinical identification and intervention of vascular cognitive impairment.

Regulation of RIP1-Mediated necroptosis via necrostatin-1 in periodontitis.

To explore the mechanism of receptor-interacting protein 1 (RIP1)-mediated necroptosis during periodontitis progression. RIP3 and mixed lineage kinase domain-like protein (MLKL) have been detected to be upregulated in periodontitis models. Because RIP1 is involved in necroptosis, it might also play a role in the progression of periodontitis. An experimental periodontitis model in BALB/c mice was established by inducing oral bacterial infection. Western blotting and immunofluorescence analyses were used to detect RIP1 expression in the periodontal ligament. Porphyromonas gingivalis was used to stimulate L929 and MC3T3-E1. RIP1 was inhibited using small-interfering RNA. Western blotting, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and enzyme-linked immunosorbent assay (ELISA) analyses were used to detect the effect of necroptosis inhibition on the expression of damage-associated molecular patterns and inflammatory cytokines. Necrostatin-1 (Nec-1) was intraperitoneally injected to inhibit RIP1 expression in mice. Necroptosis activation and inflammatory cytokine expression in periodontal tissue were verified. Tartrate-resistant acid phosphatase staining was applied to observe osteoclasts in the bone tissues of different groups. RIP1-mediated necroptosis was activated in mice with periodontitis. P. gingivalis induced RIP1-mediated necroptosis in L929 and MC3T3-E1 cells. After RIP1 inhibition, the expression levels of high mobility group protein B1 (HMGB1) and inflammatory cytokines were downregulated. After inhibiting RIP1 with Nec-1 in vivo, necroptosis was also inhibited, the expression levels of HMGB1 and inflammatory cytokines were downregulated, and osteoclast counts in the periodontal tissue decreased. RIP1-mediated necroptosis plays a role in the pathological process of periodontitis in mice. Nec-1 inhibited necroptosis, alleviated inflammation in periodontal tissue, and reduced bone resorption in periodontitis.

Wild bilberry, blackcurrant, and blackberry by-products as a source of nutritional and bioactive compounds

ABSTRACT This study aimed to investigate the nutritional and antioxidant composition of dried wild bilberry, blackcurrant, and blackberry pomaces by evaluating the proximate content, titratable acidity, color, total phenolics, total flavonoids, total anthocyanins content, and antioxidant activity. In addition, fatty acid and polyphenolic profiles were assessed using chromatographic methods. Higher fiber, fat, and mineral contents were found in blackberry as compared with bilberry and blackcurrant pomaces. The fatty acid profile revealed high concentrations of monounsaturated (16.7–18.7%) and polyunsaturated fatty acids (72.3–77.4%). The lowest n−6/n−3 ratio was found in bilberry pomace (0.90) followed by blackcurrant (1.28) and blackberry pomaces (4.26). Bilberry pomace showed the highest total phenolic (36.7 mg GAE/g dw) and total anthocyanin content (28.35 mg CGE/g dw). The phenolic profile of the bilberry pomace was dominated by ellagic acid and catechin while in blackberry and blackcurrant pomaces the major phenolic compounds were epigallocatechin and catechin, respectively. High levels of procyanidins B1 have been also quantified in bilberry and blackcurrant pomaces. Due to their healthy lipid profile and richness in antioxidant compounds, berry pomaces are good candidates as food ingredients for enhancing the functionality of food products and for contributing to the sustainability of the food industry.

USP39-mediated deubiquitination of Cyclin B1 promotes tumor cell proliferation and glioma progression

BackgroundThe elevated Cyclin B1 expression contributes to various tumorigenesis and poor prognosis. Cyclin B1 expression could be regulated by ubiquitination and deubiquitination. However, the mechanism of how Cyclin B1 is deubiquitinated and its roles in human glioma remain unclear. MethodsCo-immunoprecipitation and other assays were performed to detect the interacting of Cyclin B1 and USP39. A series of in vitro and in vivo experiments were performed to investigate the effect of USP39 on the tumorigenicity of tumor cells. ResultsUSP39 interacts with Cyclin B1 and stabilizes its expression by deubiquitinating Cyclin B1. Notably, USP39 cleaves the K29-linked polyubiquitin chain on Cyclin B1 at Lys242. Additionally, overexpression of Cyclin B1 rescues the arrested cell cycle at G2/M transition and the suppressed proliferation of glioma cells caused by USP39 knockdown in vitro. Furthermore, USP39 promotes the growth of glioma xenograft in subcutaneous and in situ of nude mice. Finally, in human tumor specimens, the expression levels of USP39 and Cyclin B1 are positively relevant. ConclusionOur data support the evidence that USP39 acts a novel deubiquitinating enzyme of Cyclin B1 and promoted tumor cell proliferation at least in part through Cyclin B1 stabilization, represents a promising therapeutic strategy for tumor patients.

A Pilot Study of Ratiometric Creatine CEST MRI Assessment of Rabbit Skeletal Muscle Energy Metabolism at 3 T.

pH MRI may provide useful information to evaluate metabolic disruption following ischemia. Radiofrequency amplitude-based creatine chemical exchange saturation transfer (CrCEST) ratiometric MRI is pH-sensitive, which could but has not been explored to examine muscle ischemia. To investigate skeletal muscle energy metabolism alterations with CrCEST ratiometric MRI. Prospective. Seven adult New Zealand rabbits with ipsilateral hindlimb muscle ischemia. 3 T/two MRI scans, including MRA and CEST imaging, were performed under two B1 amplitudes of 0.5 and 1.25 μT after 2 hours of hindlimb muscle ischemia and 1 hour of reperfusion recovery, respectively. CEST effects of two energy metabolites of creatine and phosphocreatine (PCrCEST) were resolved with the multipool Lorentzian fitting approach. The pixel-wise CrCEST ratio was quantified by calculating the ratio of the resolved CrCEST peaks under a B1 amplitude of 1.25 μT to those under 0.5 μT in the entire muscle. One-way ANOVA and Pearson's correlation. P < 0.05 was considered statistically significant. MRA images confirmed the blood flow loss and restoration in the ischemic hindlimb at the ischemia and recovery phases, respectively. Ischemic muscles exhibited a significant decrease of PCr at the ischemia (under both B1 amplitudes) and recovery phases (under B1 amplitude of 0.5 μT) and significantly increased CrCEST from normal tissues at both phases (under both B1 levels). Specifically, CrCEST decreased, and PCrCEST increased with the CrCEST ratio. Significantly strong correlations were observed among the CrCEST ratio, and CrCEST and PCrCEST under both B1 levels (r > 0.80). The CrCEST ratio altered substantially with muscle pathological states and was closely related to CEST effects of energy metabolites of Cr and PCr, suggesting that the pH-sensitive CrCEST ratiometric MRI is feasible to evaluate muscle injuries at the metabolic level. 2 TECHNICAL EFFICACY STAGE: 1.

Wheat craft beer made from AFB1-contaminated wheat malt contains detectable mycotoxins, retains quality attributes, but differs in some fermentation metabolites

Levels of aflatoxin B1 (AFB1) were measured during the production of wheat craft beer made with wheat malt contaminated with AFB1 (1.23 µg/kg). A wheat craft beer made with non-contaminated wheat malt was produced for comparison purposes. AFB1 was measured after mashing (malt after the mashing process), and in spent grain (spent grains are filtered to collect the wort - remaining sugar-rich liquid), sweet wort, green beer, spent yeast, and in beer. Physicochemical parameters (pH, titratable acidity, color parameters, total soluble solids), sugars, organic acids, alcohols, and phenolics were evaluated after mashing, and in sweet wort, green beer, and beer samples. Density and yeast counts were determined over 120 h of sweet wort fermentation every 24 h. The AFB1 levels in the final beer were 0.22 µg/L, while the spent grains and spent yeasts contained 0.71 ± 0.17 and 0.11 ± 0.03 µg/kg of AFB1, respectively. AFB1 contamination did not influence the final product's physicochemical parameters, density during fermentation, fructose, or glycerol content. Higher yeast counts were observed during the first 48 h of non-contaminated wheat craft beer fermentation, with higher ethanol, citric acid, and propionic acid contents and lower glucose, malic acid, and lactic acid contents compared with beer contaminated with AFB1. Non-contaminated wheat craft beer also had higher concentrations of gallic acid, chlorogenic acid, catechin, procyanidin A2, and procyanidin B1. AFB1 contamination of wheat malt may not affect basic quality parameters in wheat craft beer but can influence the final product's organic acid and phenolic contents. Our findings show that if wheat craft beer is made with contaminated malt, AFB1 can remain in the final product and may pose a risk to consumers.

Case 317

A 44-year-old previously healthy man with a 9-month history of progressive cognitive decline, depression, urinary incontinence, and inability to perform tasks of daily living presented to the emergency department with worsening cognitive and neuropsychiatric symptoms. He had become more distressed, and his family noticed him departing the house without closing doors, leaving water faucets running, and sending his children to school on Sundays. History taken from the patient's wife revealed that his brother had passed away in his late 30s after a slowly progressing functional and cognitive decline over the course of 5 years. No further detailed family history could be obtained. The review of systems was negative; he had no prior medical, psychiatric, or surgical history; and he denied any history of recent travel, camping, hiking, or vaccination. The patient was not taking any dietary supplements, nor was he taking any over-the-counter or prescription medication. Examination revealed vital signs were within normal limits. Neurocognitive assessment revealed a conscious, coherent, and alert patient with impaired memory and concentration. He showed poor attention, depressed mood, and restricted affect. He was unable to spell the word world forward, nor was he able to understand a request to spell it backward. The rest of the physical and neurologic examination revealed no abnormalities. Extensive laboratory work-up was conducted and included the following: toxicology screening; screening for HIV-1, HIV-2, and syphilis treponemal antibodies; COVID-19 polymerase chain reaction; and measurement of B1 and B12 levels. The results of screening were negative. Cerebrospinal fluid (CSF) assays, including CSF oligoclonal bands and CSF flow cytometry, revealed values within normal limits. CT of the brain without intravenous contrast material was performed in the emergency department to rule out acute intracranial abnormality (Fig 1). Multiplanar multisequence MRI of the brain without and with intravenous contrast material was ordered for further assessment (Figs 2-4). CT images of chest, abdomen, and pelvis were unremarkable (images not shown).

Integrated foreign languages teaching: development of learning outcomes for IT majors

Introduction. Design of descriptors for achieving objectives of foreign language education in case of non-language majors at tertiary level on the basis of the existing scales of the Common European Competences in Foreign Language (CEFR) promotes the policy of cultural diversity and multilingualism implemented by the Council of Europe. The motivation for the research lies in the fact that Russian universities lack a clear and transparent system of criteria that can be used for planning and assessing foreign language competences for non-language majors. The paper presents a developed scale of descriptors for planning learning outcomes and assessing language proficiency of undergraduate students majoring in information technologies in the context of Russian education at tertiary level. With regard to federal state educational standards for higher education, foreign language competence is described as integrative foreign language professional communicative competence which combines professional and linguistic constituents. Materials and methods. The exploration adopted combined methods of analysis of previous studies, CEFR descriptors, interviews with master and bachelor students of IT majors (Togliatti State University, Russia), and foreign language communicative needs analysis in the IT industry. The obtained data were then processed in the backward design of targeted learning outcomes following the Common European Framework of Reference for Languages format. Research results. The study elicited cross-cultural communicative needs in the industry in question, and then there were selected appropriate CEFR descriptors for their subsequent modification. The choice of the CEFR format was justified by its high adaptability and the author’s previous successful experience of elaborating such scale descriptors for automotive engineering majors. The author designed a scale for A2, B1 and B2 levels, where learning outcomes are presented as descriptors of communicative language activities with CEFR level distinctions being combined with activities involved in on-the-job cross-cultural communication. Conclusion. The initiated research should be helpful to investigate ways of devising and testing adequate sets of CEFR-based descriptors to improve the quality of university syllabi. The proposed scale of descriptors has certain limitations, therefore, further exploration directions are outlined. Keywords: integrative foreign language professional communicative competence, learning outcomes, CEFR-based descriptors, assessment, IT majors

An Investigation into Preparatory Class EFL Students’ L2 Writing Anxiety

The current paper aims to investigate foreign language (L2) writing anxiety levels of Turkish EFL students with regard to their foreign&#x0D; language writing performance, gender, age and language level. The participants were 120 preparatory class students from A1, A2, and&#x0D; B1 levels at a state university in Turkey. The data were collected through an inventory, namely The Second Language Writing Anxiety&#x0D; Inventory by Cheng (2004a), and midterm and final exam scores. According to the results of statistical procedures, it was found that&#x0D; 43.3% of the participants had high levels of foreign language writing anxiety while 34.2% of them had moderate and 22.5% of them had&#x0D; low anxiety levels. Additionally, the participants from A1 level had the highest level of writing anxiety (M=68.5). Furthermore, it was&#x0D; revealed that females had higher anxiety levels than males. The results from multiple regression analysis indicated that all variables together&#x0D; accounted for 23.3% of variance. Writing anxiety was significantly predicted by gender (β = -.33, p

Synergistic and Antagonistic Antiproliferative Effects of Ribociclib (Lee011) and Irinotecan (SN38) on Colorectal Cancer Cells.

Colorectal cancer (CRC) is one of the most common malignancies and cause of cancer-related deaths worldwide. The combination of chemotherapeutics working with different mechanisms enhances the therapeutic effects and delays the development of resistance. This study investigated the anticancer effect of the combination of ribociclib (LEE011) and irinotecan (SN38) on CRC cells. HT-29 and SW480 cells were treated with LEE011, SN38, or the combination of LEE011 and SN38. Cell viability and cell cycle distribution were analyzed. The expression of cell cycle- and apoptosis-related proteins was determined using western blot. The combination of LEE011 and SN38 elicited a synergistic antiproliferative effect on HT-29 (PIK3CAP449T mutation) cells, and an antagonistic antiproliferative effect on SW480 (KRASG12V mutation) cells. LEE011 inhibited retinoblastoma protein (Rb) phosphorylation and led to G1 arrest in HT-29 and SW480 cells. SN38 treatment caused a significant increase in the phosphorylation levels of Rb, cyclin B1, and CDC2 in SW480 cells and induced S phase arrest. Furthermore, SN38 treatment increased the phosphorylation levels of p53 and activated caspase-3 and caspase-8 in HT-29 and SW480 cells. LEE011-induced G1 arrest contributed to its synergistic antiproliferative effect with SN38 in HT-29 cells through the down-regulation of the phosphorylation of Rb. In addition, it elicited an antagonistic effect with SN38 in SW480 cells by changing the phosphorylation levels of Rb and activating caspase-8. The effects of the combination of LEE011 and conventional chemotherapy drugs on CRC depend on the chemotherapy drug and the specific gene mutation harbored by tumor cells.

Peripheral Neuropathy in a Pediatric ALL Patient (P14-9.006)

<h3>Objective:</h3> To describe the presentation and workup of a patient presenting with neuropathy found to have vitamin deficiency and anti-GD1a antibodies. <h3>Background:</h3> Peripheral neuropathy is a common finding in children with leukemia, particularly those treated with vincristine. Patients typically present with paresthesia, hyporeflexia, and weakness, often in a distal to proximal gradient. In patients receiving vincristine, symptoms typically improve within several months of stopping therapy. <h3>Design/Methods:</h3> Not applicable. <h3>Results:</h3> We present a case of a 13-year-old male from Qatar who has a history of T-cell acute lymphoblastic leukemia (T-ALL), diagnosed one year prior to presentation as well as 50lb weight loss over the past 6 months. He was admitted for cycle 3 of maintenance therapy with vincristine, prednisone, and intrathecal methotrexate. At time of admission, he reported diffuse weakness and was only able to ambulate a few steps. However, on day 7 of admission he developed complete inability to bear weight or turn in bed as well as diffuse neuropathic pain despite last vincristine dose being 6 weeks prior. Exam was notable for diffuse weakness, worse proximally and absent reflexes. Nerve conduction studies showed decreased velocity with signs of conduction block in multiple nerves suggestive of a diffuse demyelinating pathology. MRI showed no pathologic enhancement and CSF protein was normal. However, laboratory workup was notable for positive anti-GD1a antibodies as well as low levels of B1 and B6. He was started on treatment with IVIG and subsequently plasmapheresis as well as B1 and B6 supplementation. <h3>Conclusions:</h3> This case illustrates the need to consider a broad range of etiologies including vitamin deficiencies and immune mediated neuropathy when evaluating neuropathy in pediatric leukemia patients despite high prevalence of medication-induced neuropathy in this population. <b>Disclosure:</b> Dr. Wells has nothing to disclose. Dr. Kuerbitz has nothing to disclose. Dr. Fisher has nothing to disclose.

The increased expression of cytokeratin 13 leads to an increase in radiosensitivity of nasopharyngeal carcinoma HNE-3 cells by upregulating ERRFI1.

The main factors contributing to the unfavorable outcome in the clinical treatment of patients with nasopharyngeal carcinoma (NPC) patients are radiation resistance and recurrence. This study aimed to investigate the sensitivity and molecular foundation of cytokeratin 13 (CK13) in the radiotherapy of NPC. To achieve this, a human NPC cell line overexpressing CK13, HNE-3-CK13, was constructed. The effects of CK13 overexpression on cell viability and apoptosis under radiotherapy conditions were evaluated using the CCK-8 assay, immunofluorescence, and western blotting (WB). Next-generation sequencing was performed to identify the downstream genes and signaling pathways of CK13 that mediate radiotherapy response. The potential role of the candidate gene ERRFI1 in CK13-induced enhancement of radiosensitivity was investigated through rescue experiments using clone formation and WB. The effects of ERRFI1 on cell viability, cell apoptosis, cell cycle, and the related key genes were further evaluated using CCK-8, immunofluorescence, flow cytometry, quantitative polymerase chain reaction and WB. The results showed that CK13 overexpression in HNE-3 significantly inhibited cell survival under radiotherapy and promoted apoptosis marker γH2AX expression, leading to a significant increase of ERRFI1. Knockdown of ERRFI1 rescued the decreased cell viability and proliferation and the increased cell apoptosis that were caused by CK13 overexpression-mediated radiotherapy sensitization of NPC cells. In this process, EGFR, AKT, and GSK-3β were found involved. In the end, ERRFI1 was proven to inhibit expression levels of CDK1, CDK2, cyclin B1, and cyclin D1, resulting an increased G2/M cell ratio. Overexpression of CK13 enhances the radiosensitivity of NPC cells, which is characterized by decreased cell viability and proliferation and increased apoptosis. This regulation may affect the survival of HNE-3 cells by increasing the expression of ERRFI1 and activating the EGFR/Akt/GSK-3β signaling pathway, providing new potential therapeutic targets for the treatment of NPC.