Langerhans cell sarcoma with BRAF?V600E mutation and hemophagocytosis: An autopsy report

Langerhans cell sarcoma (LCS) is a rare high-grade neoplasm of langerhans cell phenotype having unambiguous malignant cytological features. We report such a rare case in a 20-year-old man who presented with dyspnea and high-grade fever. On evaluation, he had generalized lymphadenopathy, hepatosplenomegaly, and a large anterior mediastinal mass. Fine needle aspiration from the mediastinal mass and bone marrow aspirate showed numerous atypical cells, many of which showed grooved nuclei. In addition, the bone marrow showed prominent hemophagocytosis. The patient had a stormy hospital stay and succumbed to the illness. The autopsy revealed a rare multisystem involvement by LCS involving the lymph nodes, liver, spleen, lungs, and intestine, which harbored a BRAFV600E mutation and was associated with hemophagocytosis

In Vitro Antioxidant Potentiality Of Purified Anthocyanin From Floral Petals Of Wild Balsam Species

Phenolics are the largest group of phytochemicals ubiquitous in plant species with considerable interest economically. Recently, search of novel polyphenols increasingly becomes an area of intensive pharmacological research due to their multiple bioactive features such as antioxidant, antimicrobial, anti-carcinogenic, anti-viral and anti-inflammatory potentialities. Anthocyanins are flavonoid group of polyphenols, a group predominant in flowers, fruits and vegetables. The flavonoids, perhaps the unique single group of phenolics in foods, comprise a group of over 4200 C15 aromatic molecules with multiple structural patterns. The functions of anthocyanins as medicinal have been well-accepted in folk medicine throughout the world. In fact, these molecules are connected to an amazingly broad-based range of health benefits. In this juncture, the aim of this work was to evaluate the antioxidant activities of purified anthocyanin from wild balsam species. Initially, anthocyanin was extracted from floral leaves of wild balsam species and purified by chromatographic techniques. Subsequently, it was subjected to NMR and LC MS analysis. The major fractions identified were hesperidin, dimethoxy antirrhinin and trimethoxy antirrhinin. Further, the anthocyanin extracts were subjected to in vitro protocols like 2,2’-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical cation, DPPH scavenging assay, ferric reducing antioxidant power (FRAP), beta carotene bleaching assay, metal chelating and H2O2 scavenging power. Interestingly, ABTS, FRAP analyzes yielded significant results as compared to others. The data were comparable with that of synthetic antioxidants like ascorbate and catechin. Meanwhile, beta carotene and H2O2 scavenging assay showed moderate results. DPPH and metal chelating protocols displayed the values 71% and 64% respectively at 25 µg/ml concentration. This study provides model systems for the evaluation of natural antioxidants like anthocyanin. Future in vivo clinical studies are warranted to confirm the obtained data.

Bioactivity And Apoptotic Efficacy Of The Purified Terpenoid Extract From The Moss Brachythecium Buchananii (Hook.) A. Jaeger Against Mg 63 Cells

Bryophytes are primitive non vascular plants. A little is explored regarding the medicinal effects of bryophytes on carcinoma. This study investigated the biological effects of purified terpenoids from Brachythecium buchananii on selected cell lines such as HeLa, MDAMB-231 and MG63 human osteosarcoma cells and also elucidating the regu­latory signaling pathways underlying the effects of terpenoids towards caspase cascade and the antioxidant enzyme system. The cell lines were treated with various concentrations of purified terpenoid extracts interms of evaluating viability (MTT assay). Interestingly, MG63 cell lines showed poor viability as compared to other ongo cells and was subjected to further molecular evaluations. Migration and invasion assay results using wound-healing and transwell assays, respectively reveal the pro-antimetastatic potential of the purified terpenoids from the moss. The flow cytometry study substantiated terpenoid induced apoptosis in MG63 cells. Cell cycle analysis revealed the significant increase in the number of cells arrested at the S growth phase. Terpenoid extract also displayed DNA fragmentation in the cells. Western blot analysis revealed the down regulation of the anti‑apoptotic proteins Bcl‑2, pro‑caspase 3 and over expression of the pro‑apoptotic protein Bax. In addition, the caspase cascade profile of the terpenoid extract substantiated their efficacy in tumour inhibition. Thus, the overall results confirmed the biological features of terpenoid induced apoptosis in the MG63 cells.

Analysis of Bioactive Compounds by GC-MD from the Young Leaves of Tectona Grandisl.F.

The timber value of Tectona grandis has been established from decades. Teak is an exotic species cultivated along most parts of tropical regions. Anthocyanins are natural colourants which have raised a growing demand due to their extensive range of colours, innocuous and beneficial health effects. Anthocyanins belong to large group of polyphenolics - flavonoids, which are secondary metabolites synthesized by higher plants. Despite the remarkable application of anthocyanins in food, pharmaceutical and cosmetic industries, still it is not properly exploited. In the present study, the bioactive components of Tectona grandis young leaves have been evaluated using Perkin-Elmer Gas chromatography-Mass spectrometry. GC-MS analysis revealed the existence of eleven compounds. 5,9-Methanobenzocycloocten-1(2H)-one,3,4,5,6,7,8,9,10-octahydro-5,10-dihydroxy-3,3,7,7,9- pentamethyl (76.02%) and 1-naphthalenecarboxylic acid,5-[2-(3-furanyl)ethyl]decahydro-1,4a-dimethyl-6- methylene-[1R-(1.alpha.,4a.)] (13.95%). Other compounds present in minor quantities were ledol(0.92%), 3- Buten-2-one,4-(2,6,6-trimethyl-1-cyclohexen-1-yl)orionone(0.49%),9,12,15 Octadecatrienoic acid, methyl ester, (Z,Z,Z)- or Linolenic acid methyl ester (0.82%), Phytol (0.69%), Cedran-diol, 8S,14- (0.60%), Lupeol (0.71%), 3- Methoxymethyl-2,5,5,8a-tetramethyl-6,7,8a-tetrahydro-5H-chromene (3.45%) and Retinol(1.27%). This is the first report of identification of active constituents from the young leaves of Tectona grandis.

Anti-ulcer activity of arachidonic acid (PUFA) oils in different induced ulcer animal models.

Background: Ulcers of the lower part of the oesophagus, the stomach and the first part of the duodenum are also known as peptic ulcers. Peptic ulcers can affect people of any age, but they are more common as you get older. There is a focus on research for better tolerated and efficacious anti-ulcer agents. Methods: Effect of anti-ulcer activity of fish oil and Arasco oil was evaluated in different animal models of ulcers i.e. ethanol induced, water immersion and pyloric ligation techniques. The Superoxide dismutase activity in gastric tissue was also ascertained in two groups of animals. The animals received either fish oil (40 μl, PO), Arasco oil (40 μl, PO), omeprazole (20 mg/kg PO) or ranitidine (30 mg/kg PO). The gastro-protection was calculated based on ulcer index, pH and gastric juice volume. Results: The results of this study suggest that poly unsaturated fatty acid (PUFA) contained in fish oil and Arasco oil have moderate anti-ulcer activity although probably lesser in potency than the available anti-ulcer drugs like omeprazole and ranitidine. Conclusion: These results have shown that PUFA containing oils provided moderate gastrointestinal protection in all the induced ulcer models employed. Thus it can be concluded that PUFA containing oils like the Fish oil and Arasco oil have anti-ulcer properties and the mechanisms involved in these actions need to be investigated.

Cardiovascular and biochemical studies on the effects of thrombin and dabigatran and the interaction with vasopressor molecules.

Background: The effect of serine protease thrombin and its directly acting inhibitor dabigatran were evaluated on the heart rate, blood pressure, and phospholipase C (PLC) enzyme activity and the intracellular calcium levels in the platelets. Methods: Heart rate and blood pressure were estimated using electrophysiology equipment. Results: While thrombin was unable to significantly affect the heart rate and blood pressure, the inhibitor dabigatran was able to reduce the heart rate appreciably but its effects on the blood pressure were minimal. The thrombin induced increase in PLC enzyme activity, and intracellular calcium levels were attenuated by dabigatran in the platelets. The posterior pituitary hormone, vasopressin, and the adrenergic agonist noradrenaline were used to stimulate the PLC and calcium levels in platelets. Conclusion: The thrombin inhibitor, dabigatran reduces vascular oxidative stress and inflammation, improves endothelial function, and decreases atherosclerosis in rodents.

Lipoprotein associated phospholipase A2 enzyme; possible new roles and inhibition for therapeutic intervention.

Lipoprotein-Associated Phospholipase A2 (Lp-PLA2) is a 45-kDa protein of 441 amino acids encoded by the pla2g7 gene in the humans. In the blood it is associated mainly with Low Density Lipoprotein (LDL) and less than 20% is associated with High Density Lipoprotein (HDL). This enzyme is characterized by its ability to specifically hydrolyze PAF as well as glycerophospholipids containing short, truncated, and/or oxidized fatty acyl groups at the sn-2 position of the glycerol backbone. Genetic studies conducted in humans harboring an inactivating mutation at this locus suggest that loss of Lp-PLA2 function is a risk factor for inflammatory and vascular conditions. Consistently, overexpression of Lp-PLA2 has anti-inflammatory or pro-inflammatory actions and anti-atherogenic properties in animal models. This article discusses two simple techniques to estimate Lp-PLA2 activity. New therapeutic agents inhibiting the activity of Lp-PLA2 are being investigated for curative purpose.

R-Loop Hybrid Structure Formation and their Role in Health and Disease; Possible Implications for Unusual Nucleic Acid Structure Formation.

The R-loop is a stable RNA–DNA hybrid structure in which the RNA strand is base-paired with one DNA strand of a DNA duplex, leaving the opposite DNA strand single-stranded. This structure can be involved in the hypermutation and double stranded DNA breaks in mammalian immunoglobulin (Ig) genes, oncogenes and neurodegenerative disease related genes. R-loops have not been studied at the genome scale extensively. It has been shown that many oncogenes and tumour suppressors (e.g. Tp53, BRCA1, BRCA2 and Ptprd) and neurodegenerative diseases related genes (e.g. ATM, Park2, and GLDC) could be prone to significant R-loop formation. The recent findings suggest that R-loops provide a novel level of RNA–DNA interactome complexity, playing key roles in gene expression controls, mutagenesis, recombination process, chromosomal rearrangement, alternative splicing, DNAediting a R-loops have been described in vivo at the immunoglobulin class switch sequences and at prokaryotic and mitochondrial origins of replication. However, the biochemical mechanism and determinants of R-loop formation are unclear and how also they can affect epigenetic modifications needs to be elucidated. R-loop hybrid structure could be used as a novel source of prospective therapeutic targets.

Evaluation of the effect of metformin on the inhibitory effect of oxytocin on potassium chloride stimulated goat ileum.

The posterior pituitary hormone, oxytocin is expressed in the myenteric and submucous ganglia and nerve fi bers along the entire human gastrointestinal (GI) tract. The role for oxytocin in the physiology and pathophysiology of the bowel remains to be clearly elucidated. Many studies have described that oxytocin exerts stimulatory or inhibitory effects on gut functions. Recently, mRNA for oxytocin and its receptor was found throughout the entire human GI tract. In this study, we examined the responses of the posterior pituitary hormone, oxytocin on the contractile responses to KCl and the effect of metformin on these responses as it affects the glucose transport and causes monoamine release in the gut.

The Influence of Epithelial Sodium Channels in Isolated Goat Ileum Segment; Interaction with Steroids.

The contractile effect of Acetylcholine (Ach) in the isolated longitudinal ileal muscle of adult goats was studied over a varying concentration range. Ach produced a concentration dependent-response curve indicative of an interaction with muscarinic receptors in the ileum, with a maximum contraction seen at 12 μM. On the other hand, pretreatment with the ENaC blocker, Amiloride (100 μM) substantially reduced the Ach induced contractions by 67.11 %. However, pretreatment with Prednisolone (2mM) restored this effect and the relaxation induced was only 14.26 %. This change was found to be statistically significant. This study emphasizes the importance of ENaC channels in the goat intestinal smooth muscle.

ABL Tyrosine Kinases and DNA Methylation May Mediate in High Glucose Induced Hyperosmolality; Role of Target Proteins.

Phosphorylation of proteins on their threonine, serine, and tyrosine residues is one of the most commonly occurring posttranslational modifications in eukaryotic cells. Cellular phosphorylation cascades facilitates the amplification of extracellular signals following changes in environmental conditions via the ability of phosphorylated activators to modulate the expression of numerous genes. Because these reactions are rapidly reversible, they are important for the regulation of many cellular functions including signal transduction, cell division, and proliferation. Hyperosmolality can induce tyrosine phosphorylation of TonEBP/OREBP. Tyrosine phosphorylation by Abl kinases of several target proteins is a key mechanism for modulating signal transduction in hyperosmolar conditions. In this review article we discuss the role of Abl tyrosine kinases and DNA methylation during glucose induced hyperosmolality.

Application of Liquid Chromatography-Mass Spectrometry for Detecting Terlipressin Levels in Rat Plasma.

This work describes a liquid chromatography– electrospray tandem mass spectrometry method for detection of terlipressin in Wistar rat plasma in the low nano-gram range. Terlipressin is a synthetic analogue of the antidiuretic hormone arginine vasopressin and it might be used when Arginine vasopressin (AVP) is not readily available. For experimental evaluation of the pharmacokinetics of terlipressin, rats can be injected 3.0 μg/kg or 6.0 μg/kg i.v. of terlipressin and blood can be collected for Mass spectrometry characterization of terlipressin which can be performed with a high-resolution Orbitrap-based mass spectrometer.

Biosynthesis of silver nano-particles by the bacterium micrococcus luteus.

Nanotechnology is the application of science to control matter at the molecular level. It is a field that is burgeoning in the recent times, making an impact in all spheres of human life. Microorganisms play an important role in the eco-friendly synthesis of metal nanoparticles. The use of microorganisms for the synthesis of nanoparticles in the lime light of modern nanotechnology is a novel approach. Biological methods of synthesis have paved the way for the greener synthesis of these nanoparticles which can have application in biomedical sciences. In this study, we report the synthesis of nanoparticles of silver by the reduction of aqueous Ag+ by a culture of Micrococcus luteus. The formation of silver nano-particles was monitored by X-ray diffraction spectroscopy (XRD) and elucidated by transmission electron microscopy (TEM).

Is oxytocin a key hormone for gene regulation and physiological function in postmenopausal women; an inquest into possible mechanisms.

Oxytocin (OT) is a pituitary hormone and acts on the oxytocin receptor which are expressed in the brain, heart, kidney and blood vessels. OT has been implicated in its role in social bonding, and oxytocin's role in the parasympathetic nervous system includes the control of memory and learning processes and of various types of behaviour such as feeding, locomotion, as well as maternal and sexual behaviour. Oxytocin is also suggested to participate in the control of cardiovascular functions, thermoregulation, and pain threshold and fluid balance. Inefficient thermoregulation leads to hot flashes, and is associated with memory loss is a common symptom of menopause. Several studies suggest that the use of oxytocin and additional substances that amplify its effects can be used for treating weight changes, mood swings, hot flushes, somatic discomfort, dry and ulcerous mucous membranes, fissures, and bone loss during pre-menopause, menopause itself and post menopause. This review suggests its possible role in gene regulation and physiological function in post menopausal women and the possible mechanisms.

Effect of methanolic and water extract of Leucobryum bowringii Mitt. on growth, migration and invasion of MCF 7 human breast cancer cells in vitro.

Inhibitory effects of methanol and water extract of L. bowringii. on the adhesion, migration, invasion and matrix metalloproteinase (MMP) activities of MCF 7 human breast cancer cell line are reported. Cells were cultured with 10, 25, 50 µg/mL methanolic or water extract of L. bowringii. Culture medium containing 0.1% DMSO was used as a solvent control. Ultra structural analysis by electron microscopy revealed typical features of apoptosis. A remarkable dose-response parallelism was observed between methanolic extract with growth, migration and invasion of breast cancer cells. Fractionation of methanolic extract by RP-HPLC revealed a pool of phenolic acids. Hoechst 33342 staining assay reveals massive chromatin condensation and subsequent cleavage of structural components of nucleus. The results indicate that methanol extracts inhibit the growth of human breast cancer cells partially through the inhibition of metallo proteinases MMP-2 and MMP-9 activities. Methanolic extract has more anti-metastatic effects in cell based assay than water extract. Clinical application of L. bowringii extract as a bioactive chemopreventive compound may be helpful in limiting breast carcinoma invasion and metastasis.

Wound healing activity of methanolic and aqueous extracts of Plagiochila beddomei Steph. thallus in rat model.

Wound healing occurs as a fundamental response to tissue injury. Polyphenols are considered to be principle constituent to promote wound healing. Plagiochila beddomei was applied clinically by Paliyar tribes of Madurai district to treat intractable wounds. To validate the ethnotherapeutic claims of the plant in skin diseases, the effect of methanolic and aqueous extracts from P. beddomei on wound healing as well as angiogenesis was studied. 7th day and 10th day after excision wounds creation, the percent wound contraction of the extracts group was higher than that of Madecassol® (reference drug) group. On 3rd, 7th and 10th day after wounds creation, the wound healing quality of the extracts group was better than that of Madecassol® group on terms of granulation formation and collagen organization. On 3rd day after wounds creation, the micro vessel density and vascular endothelial growth factor expression of methanolic extracts group was higher than that of Madecassol® group. Phytochemical analysis of the extracts showed the presence of flavonoids, saponins, tannins and phenols. The results show that P. beddomei extract has potent wound healing property probably resulting from the remarkable angiogenic activity.

Application of western blotting technique for evaluating the expression of vasopressin receptors in the heart cells; importance in the cardiovascular system.

Vasopressin, a posterior pituitary hormone is responsible for water reabsorption by the kidneys and maintenance of cardio-vascular homeostasis. Vasopressin receptors are characterized as VR 1 (V1a), VR2 (V2), and VR3 (V1b). VR1, which is abundant in vascular smooth muscles, causes vasoconstriction by increasing intracellular calcium via the phosphatidylinositol bisphosphonate pathway and a positive inotropic effect in cardiac muscle. VR2 has also been shown to be expressed in the heart. There is emerging role for vasopressin receptors in health and disease. This study describes the application of Western blotting to elucidate the importance of vasopressin receptors in the heart cells.

Central nervous regulation of spleen function: New insights from animal studies.

The spleen is located in the upper left quadrant of the abdomen. It has two main functions that is acting as part of the immune system and as a filter. The spleen has a thin connective tissue capsule from which short septa extend inwards. These septa are, in turn, connected to a complex reticulin framework.There are two distinct components of the spleen, the red pulp and the white pulp. The red pulp consists of large numbers of sinuses and sinusoids filled with blood and is responsible for the filtration function of the spleen. The splenic venomotor fibres join the left phrenic nerve in the mid-cervical region. Coursing with it as non-medullated fibres, they eventually perforate the diaphragm, where for a time they accompany the inferior phrenic artery. Deviating towards the celiac ganglion, they next join company with the splenic vein, and are eventually distributed to localised parts of the vein. This review article evaluates the conventional knowledge and points to new insights into neural regulation of spleen.

Investigation of seizure activity after cyclic nucleotide phosphodiesterase inhibition with second messenger and calcium ion channel inhibition in mice.

The role of PDE-4 inhibitor etazolate, was evaluated in the presence of PDE-7 inhibitor, BRL-50481, in animal models of epilepsy. Seizures were induced in the animals by subjecting them to injection of chemical convulsants, Pilocarpine, Kainic acid (KA) and maximal electroshock (MES). The combination of etazolate and BRL50481 treated mice showed a significant (P<0.001) quick onset of action, jerky movements and convulsion when compared to gabapentin. The combination of etazolate and sGC inhibitor, methylene blue (MB) treated mice showed a significant (P<0.001) delay in onset of action, jerky movements and convulsion when compare to gabapentin as well as against the combination of etazolate with BRL 50481.The present study mainly highlights the individual effects of etazolate and combination with BRL-50481 potentiates (P<0.001) the onset of seizure activity against all models of convulsion. The study mainly comprises the onset of seizures, mortality/recovery, percentage of prevention of seizures (anticonvulsant) and total duration of convulsive time. The total convulsive time was prolonged significantly (P<0.05 and P<0.01) in combination of methylene blue with etazolate treated (28.59% and 35.15 %) groups, compared to DMSO received group (100%) in the MES model. In the same way, the combination of calcium channel modulator (CCM) and calcium channel blocker (CCB) amiodarone and nifedipine respectively, with etazolate showed a significant (P<0.001) delay in onset of seizures, compared to DMSO and etazolate treated groups in all models of epilepsy. This confirms that both CCM and CCB possess anticonvulsant activity. Finally, the study reveals that identification of new cAMP mediated phosphodiesterases family members offers a potential new therapy for epilepsy management in future.

Vasopressin hormone receptor density and gene expression in lithium induced polyuria rats.

Lithium, a drug frequently used for treatment of affective disorders, is known to cause a vasopressin resistant state, leading to polyuria and polydipsia. It has been suggested that lithium interacts with the renal V2-vasopressin receptor and makes the receptor dysfunctional. Detailed studies on the influence of lithium on the AVP receptor, however, have so far been difficult due to the lack of a suitable radioligand with high specific activity and high affinity. In this study, the effect of Lithium carbonate (20mg/kg i.p) on vasopressin receptor binding in the kidney and brain and the effect on mRNA expression was determined. The results of this study suggest that there was significant change in the receptor binding and gene expression in tissues of polyuria rats as compared to the control rats.