Symmetrical and asymmetrical fluorinated phenyltriazolyl-thiodigalactoside derivatives have been synthesized and evaluated as inhibitors of galectin-1 and galectin-3. Systematic tuning of the ...phenyltriazolyl-thiodigalactosides’ fluoro-interactions with galectin-3 led to the discovery of inhibitors with exceptional affinities (K d down to 1–2 nM) in symmetrically substituted thiodigalactosides as well as unsurpassed combination of high affinity (K d 7.5 nM) and selectivity (46-fold) over galectin-1 for asymmetrical thiodigalactosides by carrying one trifluorphenyltriazole and one coumaryl moiety. Studies of the inhibitor–galectin complexes with isothermal titration calorimetry and X-ray crystallography revealed the importance of fluoro-amide interaction for affinity and for selectivity. Finally, the high affinity of the discovered inhibitors required two competitive titration assay tools to be developed: a new high affinity fluorescent probe for competitive fluorescent polarization and a competitive ligand optimal for analyzing high affinity galectin-3 inhibitors with competitive isothermal titration calorimetry.
Background and aim: Flavonoid rich plant Tephrosia purpurea (T. purpurea), commonly known as Sarpunkha has been used in traditional systems of medicine to treat diabetes mellitus. However, its ...effectiveness in promoting regeneration of pancreas in diabetes has not been investigated. Therefore, the present study was undertaken to evaluate pancreatic β-cells regeneration, antioxidant and antihyperlipidemic potentials of T. purpurea leaves extract, its fractions and main constituent Rutin in diabetic rats. Experimental procedure: The leaves extract and its fractions were first screened for acute and sub-chronic antidiabetic activity in a dose range of 250-500 mg/kg orally. Further, fractions with potent antidiabetic activity were screened for pancreatic β-cells regeneration activity using histopathological studies and morphometric analysis, which was followed by estimation of biochemical parameters. Results and conclusion: The most significant antidiabetic, pancreatic regeneration and antihyperlipidemic activity was exhibited by n-butanol soluble fraction of ethanol extract at the dose level of 500 mg/kg. Histopathology revealed that treatment with this fraction improved the β-cell granulation of islets and prevented the β-cells damage which was further confirmed by morphometric analysis. Thus, the present study validated the traditional use of T. purpurea plant in the treatment of diabetes, which might be attributed to pancreatic β-cells regeneration potential of its active constituent Rutin. Taxonomy (classification by EVISE): Traditional Medicine; Metabolic Disorder; Experimental Design; Cell Regeneration and Histopathology.
Measurable/minimal residual disease (MRD) status is suggested as a powerful indicator of clinical-outcome in T-cell lymphoblastic leukemia/lymphoma (T-ALL). Contrary to B-cell ALL, reports on T-ALL ...MRD are limited and mostly based on molecular methods, mainly from developed countries. Multicolor flow cytometry (MFC)-based T-ALL studies are very few. Clinically relevant cut-off levels and ideal time-point for MRD assessment are still inconclusive. In view of lack of T-ALL MRD data from the developing world, we evaluated the prognostic value of MFC-based post-induction (PI)-MRD assessment in T-ALL in the context of standard practice.
We included 256 childhood-T-ALL patients (age < 15 years) treated with a modified-MCP841 protocol, which uses high-dose cytarabine during consolidation, as a part of standard hospital practice. MRD was studied using 10-color 11-antibody MFC with any level of detectable disease being considered positive. Post-induction (PI)-MRD was available in all patients, and post-consolidation (PC) MRD was available mostly in PI-MRD-positive patients (
= 88).
Three years cumulative-incidence-of-relapse (3years-CIR) in PI-MRD-positive patients was inferior to negative patients (46.3% vs. 18.4%). The median relapse-free-survival (RFS), event-free-survival (EFS) and overall-survival (OS) with hazard ratio (HR) of PI-MRD-positive patients were 21.4 months vs not reached (
< 0.0001, HR-4.7), 21.6 months vs. not-reached (
= 0.0003, HR-2.01) and 37.3 months vs. not reached (
= 0.026, HR-1.64) respectively. RFS, EFS and OS of patients with PI-MRD<0.01% (
= 17) were as inferior as PI-MRD ≥ 0.01% in comparison with MRD-negative patients with HR of 4.7 (
< 0.0001), 2.45 (
= 0.0003), and 2.5 (
= 0.029), respectively. Three-years-CIR of patients with hyperleukocytosis (≥100 × 109/L) was also higher (50.5 vs. 27.6%) with inferior RFS, EFS, and OS. Among PI-MRD-positive patients, 3years-CIR, RFS, EFS, and OS of PC-MRD-positive were also inferior to that of negative patients. On multivariate analysis any-level detectable PI-MRD and hyperleukocytosis remained independently associated with inferior RFS, EFS, and OS. A combination of PI-MRD-positive status and hyperleukocytosis identified the patients with the worst clinical outcomes.
Detectable PI-MRD using MFC was found to be the strong predictive factor of inferior clinical outcome in T-ALL patients. The combination of PI-MRD status and hyperleukocytosis provides the most influential tool for the management of T-ALL in resource constrained settings from developing world.
Summary
High‐sensitivity multicolour flow cytometry (MFC)‐based B‐lymphoblastic leukaemia (B‐ALL) measurable residual disease (BMRD) assay is increasingly being used in clinical practice. Herein, we ...describe six consistently present low‐level populations immunophenotypically mimicking abnormal B‐ALL blasts in 441 BMRD samples from 301 children. These included CD19+CD123+ plasmacytoid dendritic cells differentiating from lymphoid precursors, CD10+ transitional B cells with CD10+/CD38dim‐to‐negative/CD20bright/CD45bright phenotype, CD19+ natural killer (NK) cells, CD73bright/CD10+ mesenchymal stromal/stem cells, CD73bright/CD34+ endothelial cells, and a CD34+CD38dim‐to‐negative/CD10−/CD20bright/CD45bright subset of mature B cells. We provide the proportions, comprehensive immunophenotype, and practical clues for proper identification of these low‐level populations. Knowledge regarding the presence and immunophenotype of these mimics is essential for accurate interpretation in high‐sensitivity MFC‐BMRD analysis.
T-cell/NK-cell non-Hodgkin's lymphoma (T/NK-NHL) is an uncommon heterogeneous group of diseases. The current classification of T/NK-NHL is mainly based on histopathology and immunohistochemistry. In ...practice, however, the lack of unique histopathological patterns, overlapping cytomorphology, immunophenotypic complexity, inadequate panels, and diverse clinical presentations pose a great challenge. Flow cytometric immunophenotyping (FCI) is a gold standard for the diagnosis, subtyping, and monitoring of many hematological neoplasms. However, studies emphasizing the role of FCI in the diagnosis and staging of T/NK-NHL in real-world practice are scarce.
We included T-cell non-Hodgkin's lymphoma (T-NHL) patients evaluated for the diagnosis and/or staging of T/NK-NHL using FCI between 2014 and 2020. We studied the utility of FCI in the diagnosis and subtyping of T/NK-NHL and correlated the FCI findings with the results of histopathology/immunohistochemistry. For correlation purposes, patients were categorized under definitive diagnosis and subtyping, inadequate subtyping, inadequate diagnosis, and misdiagnosis based on the findings of each technique.
A total of 232 patients were diagnosed with T/NK-NHL. FCI findings provided definitive diagnoses in 198 patients and subtyping in 187/198 (95.45%) patients. The correlation between FCI and histopathological/immunohistochemistry results (n = 150) demonstrated an agreement on the diagnosis and subtyping in 69/150 (46%) patients. Of the remaining cases, the diagnosis and subtyping were inadequate in 64/150 (42.7%), and 14/150 (9.33%) were misdiagnosed on histopathology/immunohistochemistry results. FCI provided definitive diagnosis and subtyping in 51/64 (79.7%) patients. Among these, 13 patients diagnosed with peripheral T-cell lymphoma not-otherwise-specified were reclassified (angioimmunoblastic T-cell lymphoma (AITL)-11 and prolymphocytic leukemia-2) on FCI. It corrected the diagnosis in 14 patients that were misdiagnosed (6 B-cell NHL (B-NHL), 3 Hodgkin's lymphoma, 1 acute leukemia, and 1 subcutaneous panniculitis-like T-cell lymphoma) and misclassified (3 T-NHL) on histopathological results. AITL was the commonest T-NHL misclassified on histopathological results. FCI also confirmed the definite involvement in 7/83 (8.4%) and 27/83 (32.5%) bone marrow (BM) samples reported as suspicious and uninvolved, respectively, on histopathological evaluation.
AITL was the most frequently diagnosed T/NK-NHL in this study. FCI provided a distinct advantage in detecting BM involvement by T/NK-NHL, especially in patients with low-level involvement. Overall, our study concluded that FCI plays a critical role in the diagnosis, subtyping, and staging of T/NK-NHL in real-world practice.
Carissa carandas commonly known as Karanda have a long history of use in traditional system of medicine. It is used by tribal healers of Western Ghat region of Karnataka as hepatoprotective and ...antihyperglycemic. However, no scientific data is available to validate the folklore claim. The present study has been designed to evaluate its unripe fruit for the antidiabetic activity.
In the present study, methanol extract of unripe fruits and its fractions were studied for its antidiabetic potential.
The methanol extract and its fractions were screened for antidiabetic activity in alloxan induced diabetic rats. The polyphenolic, flavonoid and flavanone contents of methanolic extract and its fractions were also determined and correlated with its antidiabetic activity.
The experimental data indicated that the methanol extract and its ethyl acetate soluble fraction has significantly lowered the elevated blood glucose levels by 48% (p<0.001) and 64.5% (p<0.001) respectively at dose level of 400mg/kg per oral after 24h as compared to diabetic control. In order to assess the role of polyphenolic components in the relevant activity, polyphenolic and flavonoid contents were determined. The polyphenolic and flavonoid content of methanol extract and its ethyl acetate soluble fraction were found to be 15.8±1.2mg and 18.55±0.34mg (gallic acid equivalent/g extract) and flavonoid content 2.92±0.03mg and 1.534±0.30mg (rutin equivalent/g extract) respectively.
The increased antidiabetic potential of ethyl acetate fraction over methanol extract is due to its partial purification achieved by fractionation which resulted in increase in degree of polymerization and segregation of secondary metabolites.
Measurable/minimal residual disease (MRD) status has been suggested as a powerful indicator of clinical‐outcome in T‐cell lymphoblastic leukemia/lymphoma (T‐ALL). Multicolor flow cytometric ...(MFC)‐based T‐ALL MRD reports are limited and traditionally based on the utilization of markers‐of‐immaturity like TdT and CD99. Moreover, studies demonstrating the multicolor flow cytometric (MFC) approach for the assessment of T‐ALL MRD are sparse. Herein, we describe an 11‐marker, 10‐color MFC‐based T‐ALL MRD method using an “approach of exclusion.”
Methods
The study included 269 childhood T‐ALL patients treated with a modified‐MCP841 protocol. An 11‐marker, 10‐color MFC‐based MRD was performed in bone marrow (BM) samples at the end‐of‐induction (EOI) and end‐of‐consolidation (EOC) time‐points using Kaluza‐version‐1.3 software.
Results
We studied EOI‐MRD in 269 and EOC‐MRD in 105 childhood T‐ALL patients. EOI‐MRD was detectable in 125 (46.5%) samples (median, 0.3%; range, 0.0007–66.3%), and EOC‐MRD was detectable in 34/105 (32.4%) samples (median, 0.055%; range, 0.0008–27.6%). Leukemia‐associated immunophenotypes (LAIPs) found useful for MRD assessment were dual‐negative CD4/CD8 (40.9%), dual‐positive CD4/CD8 (23.3%) and only CD4 or CD8 expression (35.8%); dim/subset/dim‐negative surface‐CD3 (39%), dim/subset/dim‐negative/negative CD5 (28.3%), dim/dim‐negative/negative/heterogeneous CD45 (44.7%) and co‐expression of CD5/CD56 (7.5%). EOI‐MRD‐positive status was found to be the most‐relevant independent factor in the prediction of inferior relapse‐free and overall survival.
Conclusion
We described an 11‐marker 10‐color MFC‐based highly sensitive MRD assay in T‐ALL using an approach of exclusion. The addition of CD4 and CD8 to the pan‐T‐cell markers in a 10‐color assay is highly useful in T‐ALL MRD assessment and extends its applicability to almost all T‐ALL patients.
Chenopodium album L. (C. album) is commonly known as Bathua in Hindi (Family: Chenopodiaceae). Traditionally, the plant is used as a laxative, diuretic, sedative and the infusion of the plant is used ...for the treatment of rheumatism. However, no scientific validation is available on the antirheumatic potential of the plant. In the present investigation, role of NF kappa B (NFκB) in the antiarthritic potential of extracts of aerial parts of Chenopodium album was explored and evaluated.
The defatted aerial parts of Chenopodium album were successively extracted with ethylacetate, acetone, methanol and 50% methanol to study their antioxidant capacity followed by antiarthritic potential using Complete Freund׳s adjuvant (CFA) induced arthritis model in rats. The polyphenol, flavonoid and flavanone contents of different extracts were quantified and correlated with their antioxidant capacity, antiarthritic activity and NFκB inhibition potential.
The experimental data indicated that the acetone extract of Chenopodium album (ACCA) has shown significant reduction in rat paw edema (80.13%) at dose level of 200mg/kg per oral in 21 days of this study. On 22nd day, hematological and biochemical parameters were estimated and it was observed that the altered hematological parameters (Hb, RBC, WBC and ESR), biochemical parameters (Serum creatinine, total proteins and acute phase proteins) and loss in body weight in the arthritic rats were significantly brought back to near normal level by the ACCA extract. ACCA extract significantly decreased the NFκB expression in paraventricular nucleus of hypothalamus and this effect is comparable with standard indomethacine in CFA treated rats. The polyphenolic and flavonoid content of different extracts were in the range of 14.56±0.21–42.00±0.2mg (gallic acid equivalent/g extract) and 2.20±0.003–7.33±0.5mg (rutin equivalent/g extract) respectively.
The antiarthritic activity possessed by ACCA extract can be correlated directly to its antioxidant potential, high flavonoidal content achieved by successive extraction and its capacity to inhibit the NFκB protein, as proven by immunohistochemistry study.
Display omitted
The ethanolic extract of roots of
Hemidesmus indicus R.Br. (family: Asclepiadaceae) was investigated for possible antinociceptive effect in mice. Three models were used to study the effects of ...extracts on nociception, which was induced, by acetic acid (Writhing test), formalin (Paw licking test) and hot plate test in mice.
Hemidesmus indicus R.Br. extract was administered in the dose range of 25, 50 and 100
mg/kg orally 1
h prior to pain induction. The preliminary phytochemical screening of the extract showed the presence of triterpenes, flavonoids, pregnane glycosides and steroids. Oral administration of
Hemidesmus indicus extract revealed dose-dependent antinociceptive effect in all the models for antinociception and it blocked both the neurogenic and inflammatory pain and the nociceptive activity was comparable with the reference drug. The results indicate that alcoholic extract of
Hemidesmus indicus R.Br. possesses a significant antinociceptive activity. The activity can be related with the significant phytochemicals such as triterpenes, flavonoids, and sterols reported in the root extract.
The standardized aqueous extract of leaves of
Pachyptera hymenaea (DC.) belonging to family Bignoniaceae was investigated for possible antinociceptive effect in mice. Three different models were used ...to study the effects of extract on nociception, namely acetic acid-induced writhing test, formalin test (paw licking test) and tail flick test in mice. The extract was administered 1
h prior to pain induction in the dose range of 25, 50 and 75
mg/kg orally. The extract at the given dose range reduced the acetic acid induced nociception by 44.03, 52.90 and 62.46% respectively. The extract reduced formalin effect in both the phases of experiment by 32.36, 41.94, 54.29% and 35.39, 50.17, 55.86% respectively. In the tail flick study, animals’ reaction time were increased by 22.69, 38.24 and 40.26% at the above selected doses respectively at 120
min after drug administration. Naloxone (2
mg/kg; s.c.) significantly antagonized the effect of extract in formalin and tail flick method, while partially antagonized the effect in writhing test. However caffeine completely reverted the extract effect in both the phases of formalin test.
Results of these studies revealed that the extract have significant antinociceptive activity in the used models with a possible involvement of central mechanism and adenosine system.
PDF
