Miracle-dz/UniFAR-FTUs huggingface dataset:
Development of a single-chain fragment variable fused-mutant HALT-1 recombinant immunotoxin against G12V mutated KRAS colorectal cancer cells

Background KRAS oncogenes harboring codon G12 and G13 substitutions are considered gatekeeper mutations which drive oncogenesis in many cancers. To date, there are still no target-specific vaccines or drugs available against this genotype, thus reinforcing the need towards the development of targeted therapies such as immunotoxins. Methods This study aims to develop a recombinant anti-mKRAS scFv-fused mutant Hydra actinoporin-like-toxin-1 (mHALT-1) immunotoxin that is capable of recognizing and eradicating codon-12 mutated k-ras antigen abnormal cells. One G13D peptide mimotope (164-D) and one G12V peptide mimotope (68-V) were designed to elicit antigen specific IgG titres against mutated K-ras antigens in immunised Balb/c mice. The RNA was extracted from splenocytes following ELISA confirmation on post-immunized mice sera and was reverse transcribed into cDNA. The scFv combinatorial library was constructed from cDNA repertoire of variable regions of heavy chain (VH) and light chain (VL) fusions connected by a flexible glycine-serine linker, using splicing by overlap extension PCR (SOE-PCR). Anti-mKRAS G12V and G13D scFvs were cloned in pCANTAB5E phagemid and superinfected with helper phage. After few rounds of bio-panning, a specific mKRAS G12V and G13D scFv antibody against G12V and G13D control mimotope was identified and confirmed using ELISA without any cross-reactivity with other mimotopes or controls. Subsequently, the anti-mKRAS scFv was fused to mHALT-1 using SOE-PCR and cloned in pET22b vector. Expressed recombinant immunotoxins were analyzed for their effects on cell proliferation by the MTT assay and targeted specificity by cell-based ELISA on KRAS-positive and KRAS-negative cancer cells. Results The VH and VL genes from spleen RNA of mice immunized with 164-D and 68-V were amplified and randomly linked together, using SOE-PCR producing band sizes about 750 bp. Anti-mKRAS G12V and G13D scFvs were constructed in phagemid pCANTAB5E vectors with a library containing 3.4 × 106 and 2.9 × 106 individual clones, respectively. After three rounds of bio-panning, the anti-mKRAS G12V-34 scFv antibody against G12V control mimotope was identified and confirmed without any cross-reactivity with other controls using ELISA. Anti-mKRAS G12V-34 scFv fragment was fused to mHALT-1 toxin and cloned in pET22b vector with expression as inclusion bodies in E. coli BL21(DE3) (molecular weight of ~46.8 kDa). After successful solubilization and refolding, the mHALT-1-scFv immunotoxin exhibited cytotoxic effects on SW-480 colorectal cancer cells with IC50 of 25.39 μg/mL, with minimal cytotoxicity effect on NHDF cells. Discussion These results suggested that the development of such immunotoxins is potentially useful as an immunotherapeutic application against KRAS-positive malignancies.
Reproductive Diseases and Disorders of Female Camels: An Assessment and Pathological and Bacteriological Study in Eastern Ethiopia

Camels are the most efficient domesticated animals in arid and semiarid areas of the world. In Ethiopia, they are the main livestock kept to sustain the livelihoods of pastoralists, as camels are used for milk and meat production and also for transportation. However, she-camel reproductive diseases are one of the major constraints for camel-producing communities. A cross-sectional study was conducted from November 2018 to December 2019 to identify and characterize pathological lesions and isolate possible bacteria associated with reproductive diseases and disorders in she-camels slaughtered at Dire Dawa and Babille municipal abattoirs. A total of 155 study animals were examined by recruiting all she-camels slaughtered during every abattoir visit. Overall, 562 reproductive organs, the ovaries, oviducts, uterus, and cervix, were examined through observation, palpation, and incision, and the animal- and organ-level pathological lesion prevalence were found to be 29% and 64.6%, respectively. Degenerative changes, inflammatory lesions (endometritis and salpingitis), growth disturbances (e.g., ovarian hypoplasia), and noninflammatory lesions (e.g., noninflammatory edema) were the identified pathological lesions. Occurrences of pathological changes among reproductive organs had differences where significantly the highest proportion (p = 0.00) was observed in the uteri. Of the 119 microbiological samples processed, 77.3% were positive for single or mixed bacterial genera, from which 7 different bacterial isolates and 14 other unidentified Gram-negative bacteria were detected. E. coli, Salmonella, and Staphylococcus spp. were the most frequently isolated organisms with 28.2%, 26.9%, and 12.8% frequencies, respectively. The result of the questionnaire survey showed 74% of the respondents had culled the she-camel at productive age because of poor reproductive performance associated with refused mating, abortion, and repeat breeding (poor conception). On the other hand, a majority of camel herders had poor to no information and access to modern veterinary services; nevertheless, they had good indigenous knowledge on how to manage reproductive abnormalities. Considering the importance of camels in our study area, further research on camel reproductive diseases and abnormalities with wider sample and epidemiology need to be conducted using molecular and hormonal assay techniques.
Ultrasound-Targeted Microbubble Destruction Enhances Inhibitory Effect of Apatinib on Angiogenesis in Triple Negative Breast Carcinoma Xenografts

Ultrasound-targeted microbubble destruction (UTMD) has been proven as an effective technique to assist drugs to cross the vascular wall and cell membrane. This study was aimed at evaluating the synergistic antiangiogenic and growth-inhibiting effects of apatinib (APA) and UTMD on the triple negative breast cancer (TNBC). The TNBC xenograft model was established in nude mice (n = 40) which were then randomly divided into the APA plus UTMD (APA-U) group, UTMD group, APA group, and model control (M) group (n = 10 per group). Corresponding treatment was done once daily for 14 consecutive days. The general condition and body weight of tumor-bearing nude mice were monitored. Routine blood test and detection of liver and kidney function were done after treatments. The tumor size and microcirculation were examined by two-dimensional ultrasonography (2DUS) and contrast-enhanced ultrasonography (CEUS), respectively. Then, the tumor tissues were harvested for the detection of vascular endothelial growth factor (VEGF) by immunohistochemistry and for CD31-PAS double staining to assess microvessel density (MVD) and heterogeneous vascular positivity rate. After treatments, the tumor growth and angiogenesis were significantly inhibited in the APA group and the APA-U group, and these effects were more obvious in the APA-U group. The tumor volume, CEUS parameters, VEGF expression, and MVD in the APA-U group were significantly lower than those in the APA group (P < 0.05), while there were no marked differences in the heterogeneous vascular positivity rate, body weight, and blood parameters between the two groups (P > 0.05). In the UTMD group, the tumor growth and angiogenesis were not significantly inhibited, and all the parameters were similar to those in the M group (P > 0.05). During the experiment, all mice survived and generally had good condition. In conclusion, APA combined with UTMD may exert synergistic antiangiogenic and growth-inhibiting effects on the TNBC and not increase the heterogeneous vasculature and the severity of APA-related systemic side effects.
Comparative Effects of Repetitive Odor Identification and Odor Memory Tasks on Olfactory Engagement in Older Populations – A Pilot fMRI Study

Objective This study evaluated human Blood Oxygen Level-Dependent (BOLD) responses in primary and higher-order olfactory regions of older adults, using odor memory and odor identification tasks. The goal was to determine which olfactory and memory regions of interest are more strongly engaged in older populations comparing these two odor training tasks. Methods Twelve adults 55–75 years old (75% females) without intranasal or major neurological disorders performed repetitive odor memory and identification tasks using a 3-tesla magnetic resonance scanner. Odors were presented intermittently at 10-second bursts separated by 20-second intervals of odorless air. Paired t-tests were used to compare differences in the degree of activation between odor identification and odor memory tasks within individuals. An FDR cluster-level correction of p<0.05 was used for multiplicity of tests (with a cluster-defining threshold set at p<0.01 and 10 voxels). Results Odor identification compared to memory (ie, odor identification > odor memory) contrasts had several areas of significant activation, including many of the classical olfactory brain regions as well as the hippocampus. The opposite contrast (odor memory > odor identification) included the piriform cortex, though this was not significant. Both tasks equally activated the piriform cortex, and thus when the two tasks are compared to each other this area of activation appears to be either absent (OI > OM) or only weakly observed (OM > OI). Conclusion These findings from a predominantly African American sample suggest that odor identification tasks may be more potent than memory tasks in targeted olfactory engagement in older populations. Furthermore, repetitive odor identification significantly engaged the hippocampus – a region relevant to Alzheimer’s disease – more significantly than did the odor memory task. If validated in larger studies, this result could have important implications in the design of olfactory training paradigms.
Transcriptome Analysis Reveals the Genes Involved in Growth and Metabolism in Muscovy Ducks

Muscovy ducks are among the best meat ducks in the world. The objective of this study was to identify genes related to growth metabolism through transcriptome analysis of the ileal tissue of Muscovy ducks. Duck ileum samples with the highest (H group, n = 5) and lowest (L group, n = 5) body weight were selected from two hundred 70-day-old Muscovy ducks for transcriptome analysis by RNA sequencing. In the screening of differentially expressed genes (DEGs) between the H and L groups, a total of 602 DEGs with a fold change no less than 2 were identified, among which 285 were upregulated and 317 were downregulated. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that glutathione metabolism, pyrimidine metabolism, and protein digestion and absorption processes played a vital role in regulating growth and metabolism. The results showed that 7 genes related to growth and metabolism, namely, ANPEP, ENPEP, UPP1, SLC2A2, SLC6A19, NME4, and LOC106034733, were significantly expressed in group H, which was consistent with the phenotype results. The validation of these 7 genes using real-time quantitative PCR results indicated that the expression level of ENPEP was significantly different between the H and L groups (P < 0.05). This study provides a theoretical basis for exploring the influence of the ileum on growth and metabolism in ducks.
Inspiratory Flow Parameters Through Dry Powder Inhalers in Healthy Volunteers and Patients with Chronic Obstructive Pulmonary Disease (COPD): Device Resistance Does Not Limit Use in COPD

Introduction Achieving correct inhalation technique through an inhaler to ensure effective drug delivery is key to managing symptoms in patients with chronic obstructive pulmonary disease (COPD). However, many patients struggle to use their inhalers correctly, with the resultant reduction in therapeutic benefit. Consequently, appropriate inhaler choice is important to maximize clinical benefit. The primary objective of this study was to characterize inspiratory flow parameters across two Easyhaler® inhalers and the HandiHaler® inhaler in patients with COPD and healthy volunteers. Methods In this randomized, open-label, crossover study, subjects (100 patients with COPD; 100 healthy volunteers) were trained to perform inhalations of placebo powder via two variants of Easyhaler and placebo capsules via the HandiHaler inhalers. Subjects then performed three placebo inhalations through each inhaler in a random sequence. Inspiratory flow parameters were assessed, including peak inspiratory flow (PIF), for each inhaler. A parallel sub-study was conducted in patients with COPD from the main study to assess correct use of the inhalers, patient’s preference, ability to learn to use the inhalers, and the feasibility of the In-Check Dial device to measure PIF values. Results Mean PIF rates and inspiratory volumes through the three inhalers were similar between patients with COPD and healthy volunteers, and all subjects achieved the 30 L/min PIF required for effective use of Easyhaler. Almost 70% of the 88 patients enrolled in the sub-study used the Easyhaler and HandiHaler inhalers without errors. The Easyhaler was preferred by 51% of patients, while 25% favored the HandiHaler. Teaching the use of both inhalers to almost 70% of patients was very easy. The In-Check Dial PIF values and those obtained via spirometry were strongly correlated (p<0.0001) for all three inhalers. Conclusion The respiratory performance of patients with COPD does not appear to be a limiting factor in the use of Easyhaler.
Down regulation of lactate dehydrogenase initiates apoptosis in HeLa and MCF-7 cancer cells through increased voltage-dependent anion channel protein and inhibition of BCL2

Malignant cells commonly use aerobic glycolysis for ATP production; this is known as the Warburg effect, where pyruvate is converted to lactate, by enzyme lactate dehydrogenase A (LDH-A). In this study, we have investigated the effect of inhibition of LDH-A on cells viability and identifying the mechanism of cell death in HeLa and MCF-7 cancer cells. Human cervical cancer HeLa cell line and breast cancer MCF-7 cell line were used to investigate the effect of inhibition of LDH-A by sodium oxamate on cell survival and proliferation using western blot, spectrophotometry, and immunofluorescent study. There was significant reduction in LDH-A (P < 0.001) and cell viability (P < 0.001) in a dose-dependent mode in both HeLa and MCF-7 SO-treated cancer cells. The voltage-dependent anion channel (VDAC) protein was significantly increased (P < 0.001) in association with decreased LDH-A. The proapoptotic proteins; cytochrome C (P < 0.001), BAX (P < 0.001), cleaved caspase-3 (P < 0.001), cleaved caspase-8 (P < 0.001), and cleaved caspase-9 (P < 0.001) were significantly increased in association with decreased LDH-A. While, the anti-apoptotic protein Bcl2 was significantly decreased (P < 0.001) in association with decreased LDH-A. We conclude that Inhibition of LDH-A can decrease cells viability through activation of intrinsic apoptotic pathway via increased VDAC protein and inhibition of Bcl2 as well as activation of the extrinsic apoptotic pathway through activation of caspase-8.
Yanyu Decoction for Aged Patients with Stable Coronary Artery Disease: A Systematic Review and Meta-Analysis

Background There was limited evidence of treatments aiming at aged coronary artery disease (CAD) patients. Yanyu decoction (YD) has been used as adjuvant therapy in aged patients with stable CAD and might be a new treatment worthy of recommendation for CAD patients. This study was to evaluate the combined effects of YD plus conventional pharmaceutical treatment (CPT) on senile patients with stable CAD. Methods This review was designed according to the PRISMA (Preferred Reported Items for Systematic Reviews and Meta-Analysis) recommendations. A literature search was conducted in seven electronic databases from their inception until August 2020. Primary outcomes of interest were adverse cardiovascular events, including cardiac mortality, acute myocardial infarction (AMI), and unstable angina (UA). The secondary outcomes were blood lipids and hemorheology. Studies were pooled to calculate the risk ratio or weighted mean difference and corresponding 95% confidence interval. Results Five studies recruiting 848 aged patients with stable CAD were included. Patients receiving YD as an adjuvant have fewer adverse cardiovascular events, including cardiac mortality, AMI, and UA. Besides, YD plus CPT has a better effect on reducing triglycerides, low-density lipoprotein cholesterol, and improving high-density lipoprotein cholesterol. Moreover, significant effects of YD plus CPT for reducing blood viscosity, plasma viscosity, and platelet aggregation rate were found compared with CPT alone. Conclusion YD plus CPT showed better efficacy than CPT on reducing adverse cardiovascular events and improving hemorheology and blood lipids for aged patients with stable CAD. Our findings may suggest YD as an adjuvant natural-based treatment for CAD. However, more rigorous and larger trials are essential to validate our results, and further consideration of CAD studies specific to aged patients is needed.
Malnutrition among the Elderly in Malaysia and Its Associated Factors: Findings from the National Health and Morbidity Survey 2018

Background Malaysia is predicted to become an ageing population by 2035. Malnutrition among the elderly is one of growing concern. This study aims to identify the prevalence of malnutrition and its associated factors among the elderly in Malaysia. Methods Data from the National Health and Morbidity Survey (NHMS) 2018 was analysed. This survey applied a multistage stratified cluster sampling design to ensure national representativeness. Malnutrition was identified using a validated Mini Nutrition Assessment-Short Form (MNA-SF). Variables on sociodemographic, health status, and dietary practices were also obtained. The complex sampling analysis was used to determine the prevalence and associated factors of at-risk or malnutrition among the elderly. Result A total of 3,977 elderly completed the MNA-SF. The prevalence of malnutrition and at-risk of malnutrition was 7.3% and 23.5%, respectively. Complex sample multiple logistic regression found that the elderly who lived in a rural area, with no formal or primary level of education, had depression, Instrumental Activity of Daily Living (IADL) dependency, and low quality of life (QoL), were underweight, and had food insecurity and inadequate plain water intake were at a significant risk of malnutrition (malnutrition and at-risk), while Chinese, Bumiputra Sarawak, and BMI more than 25 kgm−2 were found to be protective. Conclusions Currently, three out of ten elderly in Malaysia were at-risk or malnutrition. The elderly in a rural area, low education level, depression, IADL dependency, low QoL, underweight, food insecurity, and inadequate plain water intake were at risk of malnutrition in Malaysia. The multiagency approach is needed to tackle the issue of malnutrition among the elderly by considering all predictors identified from this study.
Use of Network Pharmacology to Explore the Mechanism of Gegen (Puerariae lobatae Radix) in the Treatment of Type 2 Diabetes Mellitus Associated with Hyperlipidemia

Rapid increases in metabolic disorders, such as type 2 diabetes mellitus (T2DM) and hyperlipidemia, are becoming a substantial challenge to worldwide public health. Traditional Chinese medicine has a long history and abundant experience in the treatment of diabetes and hyperlipidemia, and Puerariae lobatae Radix (known as Gegen in Chinese) is one of the most prevalent Chinese herbs applied to treat these diseases. The underlying mechanism by which Gegen simultaneously treats diabetes and hyperlipidemia, however, has not been clearly elucidated to date. Therefore, we systematically explored the potential mechanism of Gegen in the treatment of T2DM complicated with hyperlipidemia based on network pharmacology. We screened the potential targets of Gegen, T2DM, and hyperlipidemia in several online databases. Then, the hub targets were analyzed by performing protein-protein interaction, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment assays, and finally, the complicated connections among compounds, targets, and pathways were visualized in Cytoscape. We found that isoflavones, including daidzein, genistein, and puerarin, as well as β-sitosterol, are the key active ingredients of Gegen responsible for its antidiabetic and antihyperlipidemia effects, which mainly target AKR1B1, EGFR, ESR, TNF, NOS3, MAPK3, PPAR, CYP19A1, INS, IL6, and SORD and multiple pathways, such as the PI3K-Akt signaling pathway; the AGE-RAGE signaling pathway in diabetic complications, fluid shear stress, and atherosclerosis; the PPAR signaling pathway; insulin resistance; the HIF-1 signaling pathway; the TNF signaling pathway; and others. These active ingredients also target multiple biological processes, including the regulation of glucose and lipid metabolism, the maintenance of metabolic homeostasis, and anti-inflammatory and antioxidant pathways. In conclusion, Gegen is a promising therapeutic phytomedicine for T2DM with hyperlipidemia that targets multiple proteins, biological processes, and pathways.
Extraction and Analysis of Dynamic Functional Connectome Patterns in Migraine Sufferers: A Resting-State fMRI Study

Migraine seriously affects the physical and mental health of patients because of its recurrence and the hypersensitivity to the environment that it causes. However, the pathogenesis and pathophysiology of migraine are not fully understood. We addressed this issue in the present study using an autodynamic functional connectome model (A-DFCM) with twice-clustering to compare dynamic functional connectome patterns (DFCPs) from resting-state functional magnetic resonance imaging data from migraine patients and normal control subjects. We used automatic localization of segment points to improve the efficiency of the model, and intergroup differences and network metrics were analyzed to identify the neural mechanisms of migraine. Using the A-DFCM model, we identified 17 DFCPs—including 1 that was specific and 16 that were general—based on intergroup differences. The specific DFCP was closely associated with neuronal dysfunction in migraine, whereas the general DFCPs showed that the 2 groups had similar functional topology as well as differences in the brain resting state. An analysis of network metrics revealed the critical brain regions in the specific DFCP; these were not only distributed in brain areas related to pain such as Brodmann area 1/2/3, basal ganglia, and thalamus but also located in regions that have been implicated in migraine symptoms such as the occipital lobe. An analysis of the dissimilarities in general DFCPs between the 2 groups identified 6 brain areas belonging to the so-called pain matrix. Our findings provide insight into the neural mechanisms of migraine while also identifying neuroimaging biomarkers that can aid in the diagnosis or monitoring of migraine patients.
Synthesis of 1-azulenyl ketones by Brønsted acid mediated hydration of 1-azulenylalkynes

Preparation of 1-azulenyl ketones was achieved by metal-free hydration of 1-azulenylalkynes using trifluoroacetic acid as a Bronsted acid in good to excellent yields. The reaction was accomplished at a relatively low temperature with complete regioselectivity and compatibility of several functional groups.