Beneath the rind: A review on the remarkable health benefits and applications of the wood apple fruit.

Limonia acidissima Groff, commonly referred to as the Wood apple, is a tropical fruit belonging to Rutaceae family. Indigenous to Sri Lanka, India, and Myanmar, it is extensively cultivated throughout Southeast Asia. This fruit holds a profound historical significance in traditional medicine due to its exceptional nutritional and therapeutic attributes. Wood apple pulp is significantly abundant in ß-carotene, a precursor to vitamin A, and contains a substantial amount of vitamin B, including riboflavin and thiamine, as well as trace amounts of ascorbic acid (vitamin C). Moreover health-benefitting properties associated with L. acidissima, such as, antioxidant, hepatoprotective, antimicrobial, neuroprotective, antidiabetic, anti-inflammatory, anti-spermatogenic, analgesic, antiulcer, and antihyperlipidemic properties, are attributed to a diverse range of phytochemicals. These encompass polyphenolic compounds, saponins, phytosterols, tannins, triterpenoids, coumarins, amino acids, tyramine derivatives, and vitamins. From the findings of the various studies, it was observed that wood apple fruit shows significant anticancer activity by inhibiting the proliferation of cancer. Furthermore, wood apple finds wide-ranging commercial applications in the formulation of ready-to-serve beverages, syrups, jellies, chutneys, and various other food products. In summary, this review highlights the nutritional and phytochemical constituents of wood apple, depicts its antioxidant, anti-inflammatory, and anti-diabetic capabilities, and explores its potential in value-added product development. Nevertheless, it is crucial to acknowledge that the molecular mechanisms supporting these properties remain an underexplored domain. To ensure the safe integration of wood apple fruit into the realms of the food, cosmetics, and pharmaceutical sectors, rigorous clinical trials, including toxicity assessments, are required. These endeavors hold the potential to promote innovation and contribute significantly to both research and industrial sectors.

Exploring apple pectic polysaccharides: Extraction, characterization, and biological activities - A comprehensive review.

Apple (Malus domestica) is a popular and ancient fruit of the Myrtaceae family. Apple fruit is well-known for its great nutritional and phytochemical content consisted of beneficial compounds such as polyphenols, polysaccharides, sterols, and organic acids. Polysaccharides extracted from different parts of the apple fruit, including the peel, pomace, or the whole fruit, have been extensively studied. Researchers have investigated the structural characteristics of these polysaccharides, such as molecular weight, type of monosaccharide unit, type of linkage and its position and arrangement. Besides this, functional properties and physicochemical and of apple polysaccharides have also been studied, along with the effects of extraction procedures, storage, and processing on cell wall polysaccharides. Various extraction techniques, including hot water extraction, enzymatic extraction, and solvent-assisted extraction, have been studied. From the findings, it was evident that apple polysaccharides are mainly composed of (1 â†’ 3), (1 â†’ 6): α-ß-glycosidic linkage. Moreover, the apple polysaccharides were demonstrated to exhibit antioxidant, hepatoprotective, anti-cancer, hypoilipidemic, and enzyme inhibitory properties in vitro and in vivo. The potential applications of apple polysaccharides in the food, cosmetic, pharmaceutical, nutraceutical industries have also been explored in the present review. Overall, the research on apple polysaccharides highlights their significant potential as a source of biologically active compounds with various health benefits and practical applications.

Phytochemicals, therapeutic benefits and applications of chrysanthemum flower: A review.

Chrysanthemum is a flowering plant belonging to a genus of the dicotyledonous herbaceous annual flowering plant of the Asteraceae (Compositae) family. It is a perpetual flowering plant, mostly cultivated for medicinal purposes; generally, used in popular drinks due to its aroma and flavor. It is primarily cultivated in China, Japan, Europe, and United States. These flowers were extensively used in various healthcare systems and for treating various diseases. Chrysanthemum flowers are rich in phenolic compounds and exhibit strong properties including antioxidant, antimicrobial, anti-inflammatory, anticancer, anti-allergic, anti-obesity, immune regulation, hepatoprotective, and nephroprotective activities. The main aim of the present review was to investigate the nutritional profile, phytochemistry, and biological activities of flowers of different Chrysanthemum species. Also, a critical discussion of the diverse metabolites or bioactive constituents of the Chrysanthemum flowers is highlighted in the present review. Moreover, the flower extracts of Chrysanthemum have been assessed to possess a rich phytochemical profile, including compounds such as cyanidin-3-O-(6″-O-malonyl) glucoside, delphinidin 3-O-(6" -O-malonyl) glucoside-3', rutin, quercetin, isorhamnetin, rutinoside, and others. These profiles exhibit potential health benefits, leading to their utilization in the production of supplementary food products and pharmaceutical drugs within the industry. However, more comprehensive research studies/investigations are still needed to further discover the potential benefits for human and animal utilization.

Exploring the Chemical and Biological Potential of Jamun (Syzygium cumini (L.) Skeels) Leaves: A Comprehensive Review.

Leaves of jamun collected as agro by-produce during the cultivation of jamun is traditionally used as ayurvedic medicine to treat diabetes, gall bladder stones and other ailments. Most of the beneficial effects of jamun leaves are associated with phytochemicals found in jamun leaves such as gallic acid, tannins, mallic acid, flavonoids, essential oils, jambolin, ellagic acid, jambosine, antimellin and betulinic acid. Jamun possess curative activities like anticancer, antidiabetic, antifertility, anti-inflammatory, antidiarrheal, antimicrobial, antinociceptive, antioxidant, antiradiation, chemotherapeutic, and gastroprotective. The main goal of this review article is to provide information on the nutritional content, phytochemical composition and health promoting properties of jamun leaves. The review of literature based on the phytochemical composition and health promoting benefits of the jamun leaves, suggests that leaves can be used as potential constituent in the formulation of pharmacological drugs. From the review literature it is found that clinical, in-vivo, in-vitro studies are still required to check the health promoting effects of jamun leaves extracts on humans.

Independent Reproduction of the FLASH Effect on the Gastrointestinal Tract: A Multi-Institutional Comparative Study.

FLASH radiation therapy (RT) is a promising new paradigm in radiation oncology. However, a major question that remains is the robustness and reproducibility of the FLASH effect when different irradiators are used on animals or patients with different genetic backgrounds, diets, and microbiomes, all of which can influence the effects of radiation on normal tissues. To address questions of rigor and reproducibility across different centers, we analyzed independent data sets from The University of Texas MD Anderson Cancer Center and from Lausanne University (CHUV). Both centers investigated acute effects after total abdominal irradiation to C57BL/6 animals delivered by the FLASH Mobetron system. The two centers used similar beam parameters but otherwise conducted the studies independently. The FLASH-enabled animal survival and intestinal crypt regeneration after irradiation were comparable between the two centers. These findings, together with previously published data using a converted linear accelerator, show that a robust and reproducible FLASH effect can be induced as long as the same set of irradiation parameters are used.

Spectrum of Primary Gastric Lymphoma in India: A Series of 30 Patients.

BACKGROUND: Primary gastric lymphoma is uncommonly reported in India. We retrospectively analyzed their data in a northern Indian teaching hospital. METHODS: During a 12-year period (2000-2012), endoscopic and surgical biopsies were assessed for gastric neoplasm. Gastric biopsies from normal-looking areas, rapid urease test, and Helicobacter pylori serology were done, with 2 of 3 tests positive being considered diagnostic. We aimed to study (i) the frequency of primary gastric lymphoma among gastric neoplasm patients, (ii) its clinical profile, (iii) the diagnostic procedures needed, and (iv) the frequency of H. pylori infection among them. RESULTS: Thirty out of 324 (9.2%) patients (age 56 years, range 25-72, 73.3% male) with gastric neoplasm had primary gastric lymphoma. Presentations included dyspepsia (n = 9, 30%), gastric outlet obstruction (n = 7, 23.3%), upper gastrointestinal bleeding (n = 5, 16.7%), dysphagia (n = 4, 13.3%), malignant ascites (n = 3, 10%), and others (n = 2, 6.7%). H. pylori infection was confirmed in 7 (23.3%), 12 (40%), and 21/29 (72.4%) patients by rapid urease test and histopathology and positive anti-H. pylori IgG serology, respectively. By 2 tests, H. pylori was detected in 12 (40%) patients. Though in 60% primary gastric lymphoma was diagnosed on endoscopic biopsy, in 40%, surgical resection was required. The endoscopic and surgical diagnosis groups were comparable in age (53.4 years vs. 52.7 years), sex (male 77.8% vs. 66.7%), H. pylori infection (38.9% vs. 16.7%), presentation with dyspepsia (38.9% vs. 16.7%), organic symptoms (61.1% vs. 83.3%), and the need for repeated endoscopic biopsies before diagnosis (12.% vs. 33.3%). CONCLUSION: Primary gastric lymphoma is not uncommon (9.2%) in India, often missed on endoscopic biopsy and is associated with H. pylori infection (40%).

Aegle marmelos (L.) Correa: An Underutilized Fruit with High Nutraceutical Values: A Review.

Aegle marmelos (L.) Correa (Bael) fruit, a member of the Rutaceae family, is a major cultivated fruit plant in tropical and subtropical regions in countries of southeast Asia. Bael fruit has been a major topic for studies in recent years mainly due to its high nutritional (carbohydrates, proteins, minerals, and vitamins) value and presence of various phytochemicals, which attributed to its high medicinal value. These phytochemicals include various compounds, e.g., alkaloids, flavonoids, and phenolic acids (protocatechuic acid, gallic, and ellagic acid). The fruit extract of bael has been also an important study area for its pharmacological activities, including antidiarrheal, antioxidant, antidiabetic, hepatoprotective, radioprotective, anticancer, antiulcer properties. The current review mainly highlighted the nutritional and pharmacological activities of bael fruit. The nutritional profile and phytochemical profile were discussed in the review, along with their concentration in the fruit. Moreover, the experiments carried out in vivo and in vitro of bael fruit extracts with respect to their pharmacological activities were also discussed in the article. The recent literature based on nutritional and pharmacological values of bael fruit showed its high potential as a food and pharmaceutical product. Despite having high nutritional and pharmacological value, research related to molecular mechanisms of bael fruit is still limited, and clinical trials are needed to ensure its safety as a product in the food and pharma industries.

Onion and garlic polysaccharides: A review on extraction, characterization, bioactivity, and modifications.

Allium cepa (onion) and Allium sativum (garlic) are important members of the Amaryllidaceae (Alliaceae) family and are being used both as food and medicine for centuries in different parts of the world. Polysaccharides have been extracted from different parts of onion and garlic such as bulb, straw and cell wall. The current literature portrays several studies on the extraction of polysaccharides from onion and garlic, their modification and determination of their structural (molecular weight, monosaccharide unit and their arrangement, type and position of glycosidic bond or linkage, degree of polymerization, chain conformation) and functional properties (emulsifying property, moisture retention, hygroscopicity, thermal stability, foaming ability, fat-binding capacity). In this line, this review, summarizes the various extraction techniques used for polysaccharides from onion and garlic, involving methods like solvent extraction method. Furthermore, the antioxidant, anticancer, immunomodulatory, antimicrobial, anti-inflammatory, and antidiabetic properties of onion and garlic polysaccharides as reported in in vivo and in vitro studies are also critically assessed in this review. Different studies have proved onion and garlic polysaccharides as potential antioxidant and immunomodulatory agent. Studies have implemented to improve the functionality of onion and garlic polysaccharides through various modification approaches. Further studies are warranted for utilizing onion and garlic polysaccharides in the food, nutraceutical, pharmaceutical and cosmetic industries.

Glycolytic inhibitor 2-deoxy-d-glucose attenuates SARS-CoV-2 multiplication in host cells and weakens the infective potential of progeny virions.

AIMS: Virus-infected host cells switch their metabolism to a more glycolytic phenotype, required for new virion synthesis and packaging. Therefore, we investigated the effect and mechanistic action of glycolytic inhibitor 2-Deoxy-d-glucose (2-DG) on virus multiplication in host cells following SARS-CoV-2 infection. MAIN METHODS: SARS-CoV-2 induced change in glycolysis was examined in Vero E6 cells. Effect of 2-DG on virus multiplication was evaluated by RT-PCR (N and RdRp genes) analysis, protein expression analysis of Nucleocapsid (N) and Spike (S) proteins and visual indication of cytopathy effect (CPE), The mass spectrometry analysis was performed to examine the 2-DG induced change in glycosylation status of receptor binding domain (RBD) in SARS-CoV-2 spike protein. KEY FINDINGS: We observed SARS-COV-2 infection induced increased glucose influx and glycolysis, resulting in selectively high accumulation of the fluorescent glucose analog, 2-NBDG in Vero E6 cells. 2-DG inhibited glycolysis, reduced virus multiplication and alleviated cells from virus-induced cytopathic effect (CPE) in SARS-CoV-2 infected cells. The progeny virions produced from 2-DG treated cells were found unglycosylated at crucial N-glycosites (N331 and N343) of the receptor-binding domain (RBD) in the spike protein, resulting in production of defective progeny virions with compromised infective potential. SIGNIFICANCE: The mechanistic study revealed that the inhibition of SARS-COV-2 multiplication is attributed to 2-DG induced glycolysis inhibition and possibly un-glycosylation of the spike protein, also. Therefore, based on its previous human trials in different types of Cancer and Herpes patients, it could be a potential molecule to study in COVID-19 patients.

HIF2 Regulates Intestinal Wnt5a Expression.

Radiation therapy for abdominal tumors is challenging because the small intestine is exquisitely radiosensitive. Unfortunately, there are no FDA-approved therapies to prevent or mitigate GI radiotoxicity. The EGLN protein family are oxygen sensors that regulate cell survival and metabolism through the degradation of hypoxia-inducible factors (HIFs). Our group has previously shown that stabilization of HIF2 through genetic deletion or pharmacologic inhibition of the EGLNs mitigates and protects against GI radiotoxicity in mice by improving intestinal crypt stem cell survival. Here we aimed to elucidate the molecular mechanisms by which HIF2 confers GI radioprotection. We developed duodenal organoids from mice, transiently overexpressed non-degradable HIF2, and performed bulk RNA sequencing. Interestingly, HIF2 upregulated known radiation modulators and genes involved in GI homeostasis, including Wnt5a. Non-canonical Wnt5a signaling has been shown by other groups to improve intestinal crypt regeneration in response to injury. Here we show that HIF2 drives Wnt5a expression in multiple duodenal organoid models. Luciferase reporter assays performed in human cells showed that HIF2 directly activates the WNT5A promoter via a hypoxia response element. We then evaluated crypt regeneration using spheroid formation assays. Duodenal organoids that were pre-treated with recombinant Wnt5a had a higher cryptogenic capacity after irradiation, compared to vehicle-treated organoids. Conversely, we found that Wnt5a knockout decreased the cryptogenic potential of intestinal stem cells following irradiation. Treatment with recombinant Wnt5a prior to irradiation rescued the cryptogenic capacity of Wnt5a knockout organoids, indicating that Wnt5a is necessary and sufficient for duodenal radioprotection. Taken together, our results suggest that HIF2 radioprotects the GI tract by inducing Wnt5a expression.

Plant-Based Antioxidant Extracts and Compounds in the Management of Oral Cancer.

Oral cancer continues to be a leading cause of death worldwide, and its prevalence is particularly high in developing countries, where people chew tobacco and betel nut on a regular basis. Radiation-, chemo-, targeted-, immuno-, and hormone-based therapies along with surgery are commonly used as part of a treatment plan. However, these treatments frequently result in various unwanted short- to long-term side effects. As a result, there is an urgent need to develop treatment options for oral cancer that have little or no adverse effects. Numerous bioactive compounds derived from various plants have recently attracted attention as therapeutic options for cancer treatment. Antioxidants found in medicinal plants, such as vitamins E, C, and A, reduce damage to the mucosa by neutralizing free radicals found in various oral mucosal lesions. Phytochemicals found in medicinal plants have the potential to modulate cellular signalling pathways that alter the cellular defence mechanisms to protect normal cells from reactive oxygen species (ROS) and induce apoptosis in cancer cells. This review aims to provide a comprehensive overview of various medicinal plants and phytoconstituents that have shown the potential to be used as oral cancer therapeutics.

Neuromodulatory effects of SARS-CoV2 infection: Possible therapeutic targets.

INTRODUCTION: Although SARS-CoV-2 primarily manifests in the form of respiratory symptoms, emerging evidence suggests that the disease is associated with numerous neurological complications, such as stroke and Guillain-Barre syndrome. Hence, further research is necessary to seek possible therapeutic targets in the CNS for effective management of these complications. AREAS COVERED: This review examines the neurological complications associated with SARS-CoV-2 infections and the possible routes of infection. It progresses to illuminate the possible therapeutic targets for effective management of these neuromodulatory effects and the repurposing of drugs that could serve this purpose. To this end, literature from the year 1998-2021 was derived from PubMed. EXPERT OPINION: The neurological manifestations associated with COVID-19 may be related to poor prognosis and higher comorbidity. Identification of the key molecular targets in the brain that are potential indicators of the observed neuropathology, such as inflammatory mediators and chromatin modifiers, is key. The repurposing of existing drugs to target potential candidates could reduce the mortality attributed to these associated neurological complications.

Sphingosine kinase inhibitor, SKI-II confers protection against the ionizing radiation by maintaining redox homeostasis most likely through Nrf2 signaling.

Exposure to ionizing radiation (IR) set a series of deleterious events causing acute radiation syndrome and mortality, posing the need for a potent and safe radio-protective drug. IR induces cell death predominantly by causing oxidative stress and macromolecular damage. The pre-existing antioxidant defence machinery of the cellular system plays a crucial role in protecting the cells against oxidative stress by activation of Nrf2. The current study was undertaken to investigate the radio-protective potential of sphingosine kinase inhibitor (SKI-II), which was demonstrated to activate Nrf2 signaling. The safety and efficacy of SKI-II were evaluated with cell cytotoxicity, proliferation index, and clonogenic survival assays in different cell lines, namely Raw 264.7, INT-407, IEC-6 and NIH/3T3 cell lines. A safe dose of SKI-II was found radio-protective in all the cell lines linked with the activated antioxidant defence system, thereby resulting in the amelioration of IR induced oxidative stress. SKI-II pretreatment also significantly reduced DNA damage, micronuclei expression, and accelerated DNA repair kinetics as compared to IR exposed cells. Reduced oxidative stress and enhanced DNA repair significantly reduced apoptosis and suppressed the pro-death signaling associated with IR exposure. Furthermore, the in-vitro observation was verified in the in-vivo model (C57 BL/6). The Intra-peritoneal (IP) administration of SKI-II, 2 h before a lethal dose of IR exposure (7.5 Gy) resulted in 75% survival. These results imply that SKI-II ameliorates IR-induced oxidative stress and cell death by inducing anti-oxidant defence system and DNA repair pathways, thus strengthening its potential to be used as radiation countermeasure.

C-Phycocyanin-a novel protein from Spirulina platensis- In vivo toxicity, antioxidant and immunomodulatory studies.

A pigment-protein highly dominant in Spirulina is known as C-Phycocyanin. Earlier, in vitro studies has shown that C-phycocyanin is having many biological activities like antioxidant and anti-inflammatory activities, antiplatelet, hepatoprotective, and cholesterol-lowering properties. Interestingly, there are scanty in vivo experimental findings on the immunomodulatory and antioxidant effects of C-phycocyanin. This work is aimed at in vivo evaluation of the effects of C-phycocyanin on immunomodulation and antioxidant potential in Balb/c mice. Our results of in vivo toxicity, immunomodulatory and antioxidant effects of C-Phycocyanin suggests that C-phycocyanin is very safe for consumption and having substantial antioxidant potential and also possess immunomodulatory activities in Balb/c mice in a dosage dependent manner. C-phycocyanin doesn't cause acute and subacute toxicity in the animal model (male, Balb/c mice) studied. We have reported that C-phycocyanin exhibited in vivo immunomodulation performance in this animal model.

PTEN inhibitor bpV(HOpic) confers protection against ionizing radiation.

Exposure to Ionizing radiation (IR) poses a severe threat to human health. Therefore, there is an urgent need to develop potent and safe radioprotective agents for radio-nuclear emergencies. Phosphatidylinositol-3-kinase (PI3K) mediates its cytoprotective signaling against IR by phosphorylating membrane phospholipids to phosphatidylinositol 3,4,5 triphosphate, PIP3, that serve as a docking site for AKT. Phosphatase and Tensin Homolog on chromosome 10 (PTEN) antagonizes PI3K activity by dephosphorylating PIP3, thus suppressing PI3K/AKT signaling that could prevent IR induced cytotoxicity. The current study was undertaken to investigate the radioprotective potential of PTEN inhibitor (PTENi), bpV(HOpic). The cell cytotoxicity, proliferation index, and clonogenic survival assays were performed for assessing the radioprotective potential of bpV(HOpic). A safe dose of bpV(HOpic) was shown to be radioprotective in three radiosensitive tissue origin cells. Further, bpV(HOpic) significantly reduced the IR-induced apoptosis and associated pro-death signaling. A faster and better DNA repair kinetics was also observed in bpV(HOpic) pretreated cells exposed to IR. Additionally, bpV(HOpic) decreased the IR-induced oxidative stress and significantly enhanced the antioxidant defense mechanism in cells. The radioprotective effect of bpV(HOpic) was found to be AKT dependant and primarily regulated by the enhanced glycolysis and associated signaling. Furthermore, this in-vitro observation was verified in-vivo, where administration of bpV(HOpic) in C57BL/6 mice resulted in AKT activation and conferred survival advantage against IR-induced mortality. These results imply that bpV(HOpic) ameliorates IR-induced oxidative stress and cell death by inducing AKT signaling mediated antioxidant defense system and DNA repair pathways, thus strengthening its potential to be used as a radiation countermeasure.

Mild mitochondrial uncoupling protects from ionizing radiation induced cell death by attenuating oxidative stress and mitochondrial damage.

Ionizing radiation (IR) induced mitochondrial dysfunction is associated with enhanced radiation stimulated metabolic oxidative stress that interacts randomly with intracellular bio-macromolecules causing lethal cellular injury and cell death. Since mild mitochondrial uncoupling emerged as a valuable therapeutic approach by regulating oxidative stress in most prevalent human diseases including ageing, ischemic reperfusion injury, and neurodegeneration with comparable features of IR inflicted mitochondrial damage. Therefore, we explored whether mitochondrial uncoupling could also protect from IR induced cytotoxic insult. Our results showed that DNP, BHT, FCCP, and BAM15 are safe to cells at different concentrations range depending on their respective mitochondrial uncoupling potential. Pre-incubation of murine fibroblast (NIH/3T3) cells with the safe concentration of these uncouplers followed by gamma (γ)-radiation showed significant cell growth recovery, reduced ROS generation, and apoptosis, compared to IR treatment alone. We observed that DNP pre-treatment increased the surviving fraction of IR exposed HEK-293, Raw 264.7 and NIH/3T3 cells. Additionally, DNP pre-treatment followed by IR leads to reduced total and mitochondrial oxidative stress (mos), regulated calcium (Ca2+) homeostasis, and mitochondrial bioenergetics in NIH/3T3 cells. It also significantly reduced macromolecular oxidation, correlated with the regulated ROS generation and antioxidant defence system. Moreover, DNP facilitated DNA repair kinetics evidenced by reducing the number of γ-H2AX foci formation and fragmented nuclei with time. DNP pre-incubation restrained the radiation induced pro-apoptotic factors and inhibits apoptosis. Our findings raise the possibility that mild mitochondrial uncoupling with DNP could be a potential therapeutic approach for radiation induced cytotoxic insult associated with an altered mitochondrial function.

Interleukin-6 confers radio-resistance by inducing Akt-mediated glycolysis and reducing mitochondrial damage in cells.

Interleukin-6 (IL-6)-induced glycolysis and therapeutic resistance is reported in some cell systems; however, the mechanism of IL-6-induced glycolysis in radio-resistance is unexplored. Therefore, to investigate, we treated Raw264.7 cells with IL-6 (1 h prior to irradiation) and examined the glycolytic flux. Increased expression of mRNA and protein levels of key glycolytic enzymes was observed after IL-6 treatment, which conferred glycolysis dependent resistance from radiation-induced cell death. We further established that IL-6-induced glycolysis is activated by Akt signalling and knocking down Akt or inhibition of pan Akt phosphorylation significantly abrogated the IL-6-induced radio-resistance. Moreover, reduction of IL-6-induced pAkt level suppressed the expression of Hexokinase-2 and its translocation to the mitochondria, thereby inhibiting the glycolysis-induced resistance to radiation. IL-6-induced glycolysis also minimized the radiation-induced mitochondrial damage. These results suggest that IL-6-induced glycolysis observed in cells may be responsible for IL-6-mediated therapeutic radio-resistance in cancer cells, partly by activation of Akt signalling.

Hexokinase II inhibition by 3-bromopyruvate sensitizes myeloid leukemic cells K-562 to anti-leukemic drug, daunorubicin.

An increased metabolic flux towards Warburg phenotype promotes survival, proliferation and causes therapeutic resistance, in leukemic cells. Hexokinase-II (HK-II) is expressed predominantly in cancer cells, which promotes Warburg metabolic phenotype and protects the cancer cells from drug-induced apoptosis. The HK-II inhibitor 3- Bromopyruvate (3-BP) dissociates HK-II from mitochondrial complex, which leads to enhanced sensitization of leukemic cells to anti-leukemic drugs. In the present study, we analyzed the Warburg characteristics viz. HK-II expression, glucose uptake, endogenous reactive oxygen species (ROS) level of leukemic cell lines K-562 and THP-1 and then investigated if 3-BP can sensitize the leukemic cells K-562 to anti-leukemic drug Daunorubicin (DNR). We found that both K-562 and THP-1 cells have multi-fold high levels of HK-II, glucose uptake and endogenous ROS with respect to normal PBMCs. The combined treatment (CT) of 3-BP and DNR showed synergistic effect on the growth inhibition (GI) of K-562 and THP-1 cells. This growth inhibitory effect was attributed to 3-BP induced S-phase block and DNR induced G2/M block, resulted in reduced proliferation due to CT. Further, CT resulted in low HK-II level in mitochondrial fraction, high intracellular calcium and elevated apoptosis as compared with individual treatment of DNR and 3-BP. Moreover, CT caused enhanced DNA damage and hyperpolarized mitochondria, leading to cell death. Taken together, these results suggest that 3-BP synergises the anticancer effects of DNR in the chronic myeloid leukemic cell K-562, and may act as an effective adjuvant to anti-leukemic chemotherapy.

Atypical presentation of hairy cell leukemia: Significance of CD200 on flow cytometry.

Hairy cell leukemia (HCL) is a rare, low-grade mature B-cell neoplasm with a characteristic clinical, morphological, immunophenotypic, and more recently described molecular (BRAF p.V600E mutation) profile. It typically affects middle-aged to elderly male who present with pancytopenia and massive splenomegaly. Lymphadenopathy is usually not seen. Atypical presentations such as absence of splenomegaly and presence of lymphadenopathy and leukocytosis, a hypoplastic marrow masquerading as aplastic anemia, pose a diagnostic challenge to both clinician and pathologist. A diligent morphological examination to look for the presence of hairy cells along with flow cytometric immunophenotyping showing consistent bright expression of CD200, in addition to well-described characteristic immunophenotype, helps in correctly diagnosing the case. This can be further confirmed by the consistent presence of V600E point mutation in BRAF gene. The correct identification of HCL in these unusual clinical presentations is of utmost importance owing to a different treatment approach in these cases. We present here four such cases with atypical presentation.

Mitochondrial biogenesis and metabolic hyperactivation limits the application of MTT assay in the estimation of radiation induced growth inhibition.

Metabolic viability based high throughput assays like MTT and MTS are widely used in assessing the cell viability. However, alteration in both mitochondrial content and metabolism can influence the metabolic viability of cells and radiation is a potential mitochondrial biogenesis inducer. Therefore, we tested if MTT assay is a true measure of radiation induced cell death in widely used cell lines. Radiation induced cellular growth inhibition was performed by enumerating cell numbers and metabolic viability using MTT assay at 24 and 48 hours (hrs) after exposure. The extent of radiation induced reduction in cell number was found to be larger than the decrease in MTT reduction in all the cell lines tested. We demonstrated that radiation induces PGC-1α and TFAM to stimulate mitochondrial biogenesis leading to increased levels of SDH-A and enhanced metabolic viability. Radiation induced disturbance in calcium (Ca2+) homeostasis also plays a crucial role by making the mitochondria hyperactive. These findings suggest that radiation induces mitochondrial biogenesis and hyperactivation leading to increased metabolic viability and MTT reduction. Therefore, conclusions drawn on radiation induced growth inhibition based on metabolic viability assays are likely to be erroneous as it may not correlate with growth inhibition and/or loss of clonogenic survival.