Health and Wellness Product from Mangosteen (Garcinia mangostana L.) Rind: Bioactive Potentials

Mangosteen rind (MSR) (Garcinia mangostana L.) is a predominant component of the fruit contributing to 62% of the whole fruit. However, utilization of the same for the preparation of health products was not explored due to its sensorially less acceptable parameters. Differential extraction in different polarity solvents of MSR was done and evaluated their acceptability for product preparation.Current study thus is a detailed investigation on bioactivity profiling of MSR fraction and utilization of the same for health product preparation. Among various extracts, 70% ethanol (70%AE) yielded the maximum (15g/100g). Xanthone:Phenolic ratio was 1: 2.8, in 70%AE as opposed to hot water extract – HWE and 50% AE, which contained Xanthone:Phenolic ratio of 1:1.4/5. Higher the phenolic content obviously reduces the bitterness of Xanthones. 70% AE contained phenolics 60.08± 0.213 mg/g and xanthones 22.56± 0.317 mg/g. HPLC analysis revealed a spectrum of phenolic acids such as gallic, chlorogenic, caffeic, epicatechin, catechin and ferulic acids at various levels. Potent Free Radical Scavenging (FRS) activity, cytoprotectivity, DNA protectivity, H+K+ATPase inhibitory (PPAI) activities were observed in 70% AE. Gallic/tannic acid appear to contribute to antioxidant activity; while ferulic acid was responsible for PPAI activity in 70%AE. Among xanthones, although α- mangostin was the dominating component, gartanin, 8 deoxygartanin and 3-isomangostin contributed to FRS activity. The products were prepared from 70%AE which are sensorially acceptable. Data thus for the first time delineate the specific health beneficial role of both phenolic and xanthone constituents in MSR particularly with higher abundance of phenolics than xanthones

EPHRINB2 Knockdown in Cervical Spinal Cord Preserves Diaphragm Innervation in a Mutant SOD1 Mouse Model of ALS

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by motor neuron loss. Importantly, non-neuronal cell types such as astrocytes also play significant roles in disease pathogenesis. However, mechanisms of astrocyte contribution to ALS remain incompletely understood. Astrocyte involvement suggests that transcellular signaling may play a role in disease. We examined contribution of transmembrane signaling molecule ephrinB2 to ALS pathogenesis, in particular its role in driving motor neuron damage by spinal cord astrocytes. In symptomatic SOD1G93A mice (a well-established ALS model), ephrinB2 expression was dramatically increased in ventral horn astrocytes. Reducing ephrinB2 in the cervical spinal cord ventral horn via viral-mediated shRNA delivery reduced motor neuron loss and preserved respiratory function by maintaining phrenic motor neuron innervation of diaphragm. EphrinB2 expression was also elevated in human ALS spinal cord. These findings implicate ephrinB2 upregulation as both a transcellular signaling mechanism in mutant SOD1-associated ALS and a promising therapeutic target

Percepciones y rasgos conductuales del uso de mascarillas durante la pandemia de cólera Covid-19: estudio transversal de Kerala

Background: The first reported case of COVID in all of India was in Thrissur, Kerala. Kerala was one among the states which had high recovery rate, low death rate and slow progression which was recognized internationally in managing and controlling the COVID-19 pandemic. For efficient control of transmission of COVID 19, mask wearing was considered as primary and popular measure. Wearing mask everytime can cause discomfort and prolonged mask usage can lead to many health impacts. Mask wearing can be made comfortable and convenient, if the discomforts and health impacts of wearing mask are worked on in future. Objective: To study perceptions and behavioural traits related to mask usage in general population of Kerala, India. Methods: An online cross sectional study was conducted in Kerala among 291 adults aged 20 to 60 years with a validated questionnaire. Data was analysed with Pearson’s Chi square and Odds ratio. Results: Majority of the participants were self-motivated, though 90% of participants reported mild discomforts, yet more than 80% were inclined to proper mask wearing practice. Among the participants, 77% washed and reused masks, more than 49% changed their face touching behavior, 67 % were inspired by family and friends in wearing mask. Older participants (greater than 40 years) had higher knowledge level of mask use (98%) and wearing masks outdoors (92.8%), lesser preference for N95 mask and double mask compared to younger participants. Conclusion: Study reinforces hypothesis of mask use efficacy in controlling and reducing pandemic spread. Community perceptions and behavioural traits study can enhance planning and implementation of public health programs.Antecedentes: El primer caso notificado de COVID en toda la India se produjo en Thrissur, Kerala. Kerala fue uno de los estados que tuvo una alta tasa de recuperación, una baja tasa de mortalidad y una progresión lenta, lo que fue reconocido internacionalmente en la gestión y el control de la pandemia de COVID-19. Para un control eficaz de la transmisión de COVID 19, el uso de mascarillas se consideró una medida primaria y popular. El uso de mascarilla cada vez puede causar incomodidad y el uso prolongado de la mascarilla puede llevar a muchos impactos en la salud. El uso de mascarillas puede ser más cómodo y conveniente si en el futuro se estudian las molestias y los efectos sobre la salud del uso de mascarillas. Objetivo: Estudiar las percepciones y los rasgos de comportamiento relacionados con el uso de mascarillas en la población general de Kerala, India. Métodos: Se realizó un estudio transversal en línea en Kerala entre 291 adultos de 20 a 60 años con un cuestionario validado. Los datos se analizaron con Chi cuadrado de Pearson y Odds ratio. Resultados: La mayoría de los participantes estaban auto-motivados, aunque el 90% de los participantes informaron de molestias leves, sin embargo, más del 80% se inclinaron por la práctica adecuada del uso de mascarillas. Entre los participantes, el 77% lavaba y reutilizaba las mascarillas, más del 49% cambiaba su conducta de tocarse la cara y el 67% se inspiraba en familiares y amigos para llevar mascarilla. Los participantes de más edad (más de 40 años) tenían un mayor nivel de conocimientos sobre el uso de mascarillas (98%) y el uso de mascarillas al aire libre (92,8%), menor preferencia por la mascarilla N95 y la mascarilla doble en comparación con los participantes más jóvenes. Conclusiones: El estudio refuerza la hipótesis de la eficacia del uso de mascarillas para controlar y reducir la propagación de la pandemia. El estudio de las percepciones y los rasgos de comportamiento de la comunidad puede mejorar la planificación y la aplicación de los programas de salud pública

Poor Survival Outcomes In Myelodysplastic Syndrome Patients With Non-t(6;9) and Non-Inv(3) Balanced Chromosomal Rearrangements Are Influenced By SRSF2 Mutations

Abstract Cytogenetics is an important predictor of overall outcomes in patients with myeloid malignancies including myelodysplastic syndromes (MDS). Traditional metaphase cytogenetics can detect chromosomal abnormalities in 45-50% of MDS patients. Unbalanced chromosomal abnormalities including deletions, monosomies and others are the predominant lesions commonly encountered. The prognostic value of most of these unbalanced chromosomal changes is well established specifically -7/-7q, -5/-5q, and +8. However, the role played by balanced chromosomal rearrangements (BCR) specifically balanced translocations and inversions in MDS is less established. Most cases especially in the absence of poor prognostic chromosomal karyotypes are assigned by default to the intermediate risk group. The importance of BCR especially translocations cannot be underscored since novel fusion proteins may form during such chromosomal events that can be subsequently targeted similarly as in the prototypical disease bcr-abl positive chronic myeloid leukemia. We hypothesized that BCR are commonly found in MDS and may confer important prognostic and therapeutic impact. We studied a total of 303 MDS patients seen at the Cleveland Clinic between the years 2002-2011. There were 197 males and 106 females. The median age of the cohort is 69 years (range: 19-92). The median follow-up time is 17 months. Cytogenetic, hematologic and survival data were collected from individual patients. Responses were assessed using the International working group criteria for MDS. Categorical data were analyzed using X2 test and survival outcomes were analyzed using Kaplan-Meier method. A p-value of <0.05 was considered statistically significant. Sanger sequencing for genes relevant in MDS pathophysiology including TET2, CBL, DNMT3A, NRAS, KRAS, TP53, SF3B1, U2AF1, SRSF2, ASXL1, RUNX1, JAK2, IDH1/2 were performed according to previously published methods. Balanced rearrangements with known poor prognostic significance specifically t(6;9) and inv(3) were excluded from the analysis. A total of 23 patients with BCR (8%) were identified. The most commonly affected chromosomes are chromosomes 1 and 7 with primarily translocations including 2 cases each of t(7;8) and t(1;7); while inversions involved various chromosomes including 2, 4, 9, 10, and 11. To understand the prognostic significance of these BCR, we compared the survival outcomes of patients with BCR with those of patients with specific chromosomal defects prognostically defined using the IPSS. Patients with BCR have worse OS similar to patients with poor risk cytogenetics as defined by IPSS (BCR=7 mos vs [Good risk=23 mos, Int=15 mos, vs. poor=5 mos], p=.0013). We previously reported the prognostic importance of single nucleotide polymorphism array analysis (SNP-A) lesions in MDS. New SNP-A and acquired somatic uniparental disomy abnormalities did not alter the prognosis of patients with BCR (SNP-A: new lesions= 6.7 mos vs. no lesions= 6.7 mos; p=.57, UPD: new lesions=4.6 mos vs. no lesions=7.7 mos, p=.19). In the advent of molecular genetics, specific molecular mutations affect outcomes in MDS and related cancers. We investigated the frequency and the role played by specific molecular mutations in the outcomes of patients with BCR. Among detected somatic mutations (TET2 (9%), ASXL1 (17%), JAK2 (13%), TP53 (9%) and SRSF2 (9%)), only patient harboring mutations in the RNA splicing gene SRSF2 conferred worse outcomes in this group (MUT= 1 mo vs. WT= 9 mos, p=.001). Therapeutically, 3 patients underwent allogeneic hematopoietic cell transplant (Allo-HCT), 11 received pharmacologic therapies (Induction chemotherapy [IC]=3; hypomethylating therapy [HMT]=11) and 5 with supportive treatments. All patients who received Allo-HCT achieved a complete remission and remain alive while 2 patients who received IC and 7 patients who received HMT did not respond to treatment. In conclusion, BCR are not uncommon in MDS. Interestingly, even when excluding known poor-risk balanced translocations like t(6;9) and inv(3), they still conferred a poor survival outcome in MDS patients. RNA splicing gene mutations specifically SRSF2 is a driver of worse outcomes within this group. Patients with BCR have better outcomes when managed with allo-HCT compared to HMT and IC. Disclosures: No relevant conflicts of interest to declare

Somatic Mutations of the Breast Cancer Amplified Sequence-1 (BCAS1), a Novel Leukemogenic Driver in Myelodysplastic Syndromes with Del(20q)

Abstract Deletion of the long arm of chromosome 20 [del(20q)] is a recurrent clonal abnormality seen in myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPN), and acute myeloid leukemia (AML). Del(20q) is believed to originate in a pluripotent stem cell of both myeloid and lymphoid cells and contribute to clonal evolution by the loss/inactivation of ≥1 tumor suppressor genes. Commonly deleted regions (CDRs) of del(20q) contain genes that are overexpressed in hematopoietic progenitor cells of del(20q) patients (pts). Although associated with good prognosis when present as a sole abnormality in MDS, the natural history may vary within similar disease subtypes. Next-generation sequencing technologies have accelerated the discovery of genetic mutations implicated in MDS pathogenesis. To date, the vast majority of these mutations are neither specific to a disease subtype nor to a cytogenetic abnormality. We hypothesized that molecular mutations are the main drivers of leukemic progression in MDS with del(20q). Whole exome sequencing (WES) was performed on 6 pts with del(20q): MDS=3, MPN=3. DNA (3ug) was used for WES. 20 million reads were run on an Illumina HiSeq2000 sequencer. A bioinformatic algorithm filtered all tumor variants based on variation score (≥30) and coverage (30X). Tumor nucleotide variants were counted if unique to the tumor excluding SNPs and germline variants. We detected 42 candidate genes. Only 1 nucleotide variation was detected on the chromosome (chr) 20 of 1 high risk MDS with del(20q) pt. The alteration resulted in a proline to a glutamine substitution at position 488 of exon 11 (P488Q) of a gene called Breast Cancer Amplified Sequence-1 (BCAS1). BCAS1 gene is located in chr 20q13.2 region. BCAS1 overexpression has been implicated in aggressive breast and colon cancer subtypes but the mechanisms for this overexpression have not been clarified. Of note, BCAS1 mutations have not been previously identified in any hematologic malignancy. A larger cohort of pts: N=395; AML=70, MPN=205, MDS=90, other bone marrow failure diseases=30, and hematologic cell lines were tested for BCAS1 mutations. We found recurrent BCAS1P488Q mutation in 10/56 (18%) of MDS with del(20q) pts who evolved to high risk disease and AML (sAML). All BCAS1 mutant (BCAS1MUT) pts have an associated del(20q) abnormality. BCAS1MUT pts have a median age of 67 yrs (range, 45-87); male/female=9:1, had more leukopenia (3 vs. 11 x109/L, P=.04), and neutropenia (1.3 vs. 8.3 x109/L, P=.04) compared to other del(20q) pts without mutation. Metaphase cytogenetics, FISH and SNP-A showed that the CDRs of the 20q region of all BCAS1MUT pts did not contain the 20q13.2 region. Screening for 12 genes relevant to MDS pathophysiology showed that BCAS1MUT pts harbored unfavorable genetic mutations: SRSF2 (1/10), TP53 (2/10) and U2AF1 (3/10) with 1 pt having both TP53 and U2AF1. BCAS1P488Q was mutually exclusive in 4 pts. BCAS1MUT had a dismal prognosis compared to BCAS1WT (PFS: 5.1 vs. 15.9 mos, P=.03). Bioinformatic analysis of publicly available gene expression data showed increased BCAS1 mRNA level in AML, CML and in the K562 cells. RT-PCR found a higher (5-fold) BCAS1 mRNA level in the index case compared to one WT pt and no expression in healthy subjects. K562 and CML primary cells (N=6) were used as positive controls. Presence of BCAS1 protein was confirmed by immunohistochemistry and western blotting. CGH revealed that 10% of breast tumors have copy-number gains of a 1-Mb region of 20q13 where BCAS1 maps. Amplification of this region was seen in aggressive subtypes of squamous cell carcinoma. We applied FISH analysis for BCAS1 finding no amplification of the 20q13 region in MUT vs WT (2 vs 2) pts suggesting that other mechanisms possibly other genetic mutations lead to BCAS1 overexpression in sAML. In the K562 cells, 3 copies of BCAS1 were identified which may explain increased BCAS1 expression in these cells. In sum, recurrent BCAS1 mutations are found specifically in MDS with del(20q) pts who transform to AML. They are exclusively found at the time of AML transformation. mRNA and protein overexpression are observed in BCAS1MUT pts and lead to inferior survival outcomes and poor response to chemotherapy. This is the first genetic mutation solely found in a specific MDS cytogenetic subset and may represent a novel mechanism for leukemogenesis. Disclosures No relevant conflicts of interest to declare

Molecular Characterization Of Myeloid Neoplasms Harboring Isochromosome 17q Abnormality

Abstract Isochromosome 17q [i(17q)], a poor prognostic cytogenetic abnormality is a product of the breakage or inappropriate division of the pericentromere leading to the duplication of the long and loss of the short arm of chromosome 17. The region of the breakpoints maps at 17p11, a region encompassing a key tumor suppressor gene: TP53. I(17q) are detected in myelodysplastic/ myeloproliferative neoplasms (MDS/ MPN), chronic myeloid leukemia (CML), and acute myeloid leukemia (AML). This abnormality can occur as a sole structural abnormality or in combination with other chromosomal defects. The presence of i(17q) is associated with poor therapeutic response, disease progression, and an unfavorable clinical outcome. Elucidation of the molecular architecture of patients (pts) carrying i(17q) may lead to better understanding of disease biology and development of novel compounds that can target this disease. We selected 11 pts with i(17q) to characterize their genomic differences. We applied whole exome sequencing (WES) in order to define latent molecular defects explaining the clinical phenotype of this disease. The index case was a male MDS/MPN pt with isolated i(17q), 27% RS, hypercellular bone marrow (BM), mild splenomegaly, and atypical megakaryocytes. The pt developed 7% BM blasts without clinical response to growth factors. Molecularly this pt was a wild type SF3B1, a gene frequently mutated in RARS-T and associated with lower transformation rate to leukemia, better survival, and good/intermediate risk cytogenetic abnormalities. WES was performed on 2 ug of total DNA extracted from BM cells. Non-clonal CD3+ cells were used as source of germ-line control. Twenty-millions reads were run on an Illumina HiSeq2000 sequencer. Using a stringent bio-informatic algorithm developed in house, all variants were filtered based on a variation score (>=30) and a coverage (30X) and the tumor nucleotide variation analysis was performed for each pair (tumor vs. germ-line), where only the variants unique to the tumor were retained. Variants were ultimately filtered in order to exclude SNPs by an in-house annotation and importing the hg19 SNP135. We detected 65 unique candidate genes. Four genes were confirmed to be somatic: 3 were novel: ZFP42 (4q35.2), P4HTM (3p21.31), and VPRBP (3p21.2) and 1 includes the newly discovered SETBP1 (18q12.3) gene. Three variants detected on the chromosome 17 had a wild type configuration. The subsequently genotyped all the pts (MDS/MPN/-U 3; AML 4; RCMD 1; CML 1; RAEB-1 2; mean age: 68 years; male/female: 8/3; i(17q)/other abnormalities:3/8) for the above genes and for a panel of genes known to be mutated in MDS/MPN and other diseases in order to find any genetic association explaining the disase phenotype. We applied Sanger sequencing to DNA derived from BM/peripheral blood cells (BM/PB:7/4) for the following genes and respective exons: TP53 (all exons), SF3B1 (13-16), SRSF2 (1-2), U2AF1 (2 and 6), TET2 (all exons), DNMT3A (18-23), IDH1/2 (4), CBL (8-9), N/KRAS (1-2), ASXL1 (12), JAK2 (12 and 14), EZH2 (16, 18 and 19), MPL (exon 10), BCAS3 (12, 15 and 16), FLT3 (11 and 17), and CSF3R (13,14, and 17). In total, we found 16 heterozygous missense mutations and 1 tandem duplication. We found somatic mutations in ZFP42, P4HTM, and VPRBP in 1 pt. The index case reported a mutation in SETBP1 and SRSF2. SF3B1 was detected as a sole abnormality in 1 patient. Of note, the patient with SF3B1 mutation (K700E) had 50% RS and achieved a complete hematologic remission after decitabine therapy. The most frequent mutations were found in SETBP1 and SRSF2. SETBP1 was found to be mutated in 4/11 (36.3%) pts (D868N, I871T, and G870S was common in 2 pts) while SRSF2 mutations (P95H/R) were found in 3/11 (27.2%) pts. Three pts showed concomitant SRSF2 and SETBP1 mutations. NRAS (G12D) was mutated in 1 pt and associated with SRSF2 and SETBP1 mutations. One pt showed mutations in TET2, JAK2, and TP53. Of note, this pt did not respond to treatment. One pt with MDS/MPN showed a mutation in CSF3R (Q741X), a novel gene discovered in chronic neutrophilic leukemia and atypical CML. The pt also has monosomy 7 and i(17q) abnormality. FLT3-ITD was found in 1 pt. As of last follow-up, only 2 pts remain alive. In sum, we found that poor risk molecular mutations in SRSF2 and SETBP1 are frequently found in i(17q) myeloid malignancies and may be the drivers of poor outcomes in this disease. Disclosures: No relevant conflicts of interest to declare