When molecular docking simu lation among tylophorine ligand and VEGFR2 protein was analyzed, it was identified that tylophorine has bound at slightly distinct location towards N terminal domain from original bound ligand 42Q with seven. 00 Kcal/mol binding af finity within the ATP binding pocket. There are actually five amino acids i. e, Lys868, Leu870, His879, Leu882 and Leu912 are actively involved in the binding of tylophorine. His879 is definitely an active amino acid with the ATP binding pocket has participated in hydrogen bond with tylophorine. Rest amino acids are hydrophobic in nature and have manufactured robust ? ? bonds using the ligand. Consequently hydrophobic interaction is far more dominant than hydrogen and electrostatic interaction in tylophorine VEGFR2 complex.
When structure of tylophorine was inspected, it’s observed that its core construction has produced up with three fused benzene rings that are also hydrophobic nature suggesting, it might be motive for dominancy selleck chemical of hydrophobic interaction. Such binding pattern of tylophorine inside VEGFR2 may perhaps prohibit the binding from the ATP at its binding pocket and in this way it has offered a path for devel opment of tiny all-natural inhibitors. Discussion The current examine demonstrated that tylophorine exhibited anti angiogenic pursuits in vivo and suppressed crucial ways involved with angiogenesis together with proliferation, migration, invasion, tubulogenesis and expression of pro MMP2 as detected by gelatin zymography in endothelial cells. By dir ectly blocking VEGFR2 phosphorylation and activation, tylophorine inhibited VEGFR2 kinase activity too as suppressed VEGFR2 signaling pathway in vitro.
Supporting evidences regarding in vivo anti angiogenesis results inhibitor CP-690550 of tylophorine then came from sponge implant angiogenesis model and Ehrlich ascites carcinoma tumor model. Tylophorine significantly inhibited blood vessels formation in sponge implant assay and considerably suppressed tumor growth accompanied by reduction in microvessel density in tumor tissues. Our review provides a novel and mechanistic insights in to the mechanism by which tylophorine has an effect on the numerous facets of vascular endothelial angiogenic signal ing as a result of VEGFR1 and VEGFR2. Phosphorylated Tyr1175 of VEGFR2 mediates activation in the mitogen activated protein kinase/ERK cascade and was proven to contribute to cell proliferation in endothelial cells. Src household kinase is considerably involved with VEGF induced angiogenesis in vitro and in vivo. Other signaling molecules which have been involved in VEGF induced migration by means of VEGFR2 include FAK and its substrate paxillin, which are participated in focal adhesion throughout cell migration.

Success have been presented at ASCO 2010 displaying that carboplatin plus PLD is just not superior to carboplatin plus paclitaxel when it comes to PFS, the median PFS was 19 and 16. eight months during the former as well as lat ter arms, respectively. Having said that, provided the observed con fidence interval along with the unique toxicity profile it has been proposed that carboplatin plus PLD may very well be con sidered an different to conventional treatment. Quite a few randomized trials are performed in pla tinum delicate individuals. A multicenter phase III study, recently published, the Calypso review, has com pared efficacy and safety of PLD carboplatin and carbo platin paclitaxel in 976 relapsed platinum sensitive ovarian cancer sufferers. The trial showed superiority of the experimental arm when it comes to PFS.
The security profile of PLD carboplatin seems remarkably unique from that of carboplatin plus paclitaxel. The PLD carboplatin mixture was associated using a larger incidence of anemia and thrombocytopenia in addition to a larger incidence of stomatitis and cutaneous toxicity. Notably, having said that, the PLD carbo platin mixture was connected with a knockout post a really low inci dence of hair reduction and neurotoxicity in contrast in between the 2 arms was located regarding response price. A single intriguing observation of this trial was in PLD car or truck boplatin arm in contrast to carboplatin paclitaxel there was the reduction in the price of hypersensitive response Therapeutic Approaches in Epithelial Ovarian Cancer and this is often significant informa tion given that hypersensitive reactions are reported while in the general practice in individuals taken care of with carboplatin as much as 25%.
Therapy of clear cell type of EOC Even though clear cell kind is categorized in Kind I ovarian cancer, it’s known to demonstrate relatively sturdy resistance to carboplatin and paclitaxel routine and so poor prognosis compared to serous adenocar cinoma, particularly in advanced phases. Previously Sugiyama MGCD0103 Mocetinostat et al. investigated clinical qualities of clear cell carcinoma with the ovary and showed that individuals with CCC were significantly far more likely to have FIGO Stage I illness than had been individuals with SAC. However, a higher recurrence rate was mentioned in these sufferers with Stage IC CCC plus the survival prices for anyone stage IC CCC patients have been lower than individuals for patients with SAC. Also, the 3 year and five yr survival costs for Stage III CCC patients have been drastically reduced in contrast with Stage III SAC patients. Enomoto et al. demonstrated that clear cell or muci nous carcinoma histologic sort didn’t react on the carboplatin paclitaxel combination chemotherapy. Considering individuals past reports, choice chemotherapy regimens or novel treatment method for clear cell and mucinous carcinoma need to be investigated.

2 nM and 186. 7 nM. There was no major distinction among the response to SNS 032 as well as traits of AML individuals. How ever, a smaller fraction of your specimens was rela tively resistant to SNS 032 mediated cell death. Also, a significant lower inside the number of colony formation was observed while in the primary blasts obtained from 4 individuals with newly diagnostic AML, but not within the bone marrow cells from healthier volunteers. SNS 032 induced apoptosis and inhibited not just phosphorylation of RNA Pol II but additionally phosphorylation of mTOR and its downstream targets Former research showed that induction of apoptosis can be a crucial action for SNS 032 induced cell death in AML and CML. We thus evaluated the result of SNS 032 on apoptosis of AML cell lines. Cells were handled with increas ing doses of the drug for 24 h, then apoptotic cells have been determined by annexin V FITC.
The 50% successful concen tration of KG one and HL 60 cell lines was 192. 2 and 194. eight nM, respectively. In contrast, read this article HEL cells were resistant to SNS 032 induced apoptosis. There was tiny cell death at 24 h right after SNS 032 treatment method, even at concentration of 200 nM. To examine the cell cycle effects, HL 60 cells had been cultured with SNS 032 or Rapamycin, respect ively, and cell cycle analysis was performed. The cells exposed to SNS 032 showed accumulations of cells in G1 phase, consistent with prior reports that exhibiting that SNS 032 induces a cell cycle arrest. The greater percentages of cells during the G1 phases have been also observed in HL 60 cells taken care of with Rapamycin. Subsequent, we set out to unravel the molecular mechanism of action of SNS 032. On western blot evaluation, we observed that SNS 032 dose dependently decreased phosphorylation of RNA pol II at Ser2 and Ser5 in KG one and HL 60 cells following 6 h of incubation.
They are constant together with the former report. Interestingly, we uncovered that SNS 032 strongly inhibited phosphorylation of mTOR on Ser2448, a marker for mTORC1 exercise, also as phosphorylation of mTOR protein discover this on Ser2481, a marker to the presence of mTORC2 complexes. The ac tivity of mTORC1 and mTORC2 in HL 60 and KG 1 cells was completely inhibited through the treatment method with 200 and 400 nM SNS 032 accompanied by slight degradation of protein expression of mTOR. The downregulation of endogenous levels of mTOR protein phosphorylated at Ser2448 was also confirmed from the taken care of HL 60 cells using ELISA assays. To check the impact of SNS 032 on unrelated signaling pathways, immunoblotting examination was performed. The addition in the drug didn’t suppress extracellular signal regulated kinase Thr202/ Tyr204 phosphorylation, p38 mitogen activated protein kin ase Thr180/Tyr182 phosphorylation in HL 60 cells, and also did not lessen signal transducer and activa tor of transcription 5 Tyr694 phosphorylation and STAT3 Tyr705 phosphorylation.

Both floating cells and trypsinized adherent cells were collected and mixed for examination. Cells had been fixed by dropwise addition into ice cold etha nol and stored at 20 overnight. Cells were then pel leted, washed, and stained for 1 hour with 50 ug/ml propidium iodide in PBS containing 0. one mg/ml ribonu clease A and 0. 05% Triton X 100. Right after gating to exclude debris, the DNA content material was measured making use of a LSR II erismodegib Smoothened Inhibitors movement cytometer. Information had been analyzed with ModFit LT computer software. Data are represented as percent reside cells of two independent experiments. Plasmid packaging and steady cell line generation HEK293T cells had been plated at 5. 5 ? 106 in a 10 cm dish in 10 ml of 10% MEM and permitted to adhere in excess of night. The next day, the HEK293t cells have been co transfected together with the pLEmiR non silencing turbo red fluorescent protein vector construct and the trans lentiviral packaging mix and pLEX transfer vector utilizing the TransLenti Viral pLEX packaging sys tem, following the manufacturers directions.
Virus was harvested 48 hrs submit transfection and stored at 80 C. TNBC TRAM-34 cell lines had been plated at 70% confluence in 10 cm dishes with 10 ml of 10% MEM and permitted to adhere over night. The following day, cells had been transduced with virus containing the pLEmiR tRFP vector in serum free media following the producers proto col. Immediately after 4 hours, the transduction media was removed and replaced with 10% MEM. Immediately after 24 hours, cells had been treated with puromycin to pick for vector expression. The resultant stable trans fectants had been designated as MDA MB 231 tRFP and BT 549 tRFP. Animal xenograft studies Xenograft tumor studies were performed as previously described. In short, CB 17/SCID female mice had been obtained from Charles River Laboratories.
The animals were permitted a time period of adaptation within a sterile and patho gen absolutely free environment with meals and water ad libitum. MDA MB 231 tRFP and BT 549 tRFP cells had been har vested while in the exponential development phase utilizing a PBS/ ethylenediaminetetraacetic acid solution and washed. Viable cells in 50 ul of sterile PBS sus pension had been mixed with a hundred ul decreased growth component Matrigel and injected bilaterally to the inguinal mammary unwanted fat pad. On day three post cell injection, mice were randomized into therapy groups of five mice each. Beginning on day 14 publish cell injection, animals obtained intraperitoneal injections from the corre sponding drug treatment on a five day on and two day off schedule for 28 days. Tumor size was measured by using a digital caliper and calculated utilizing the formula 4/3?LS2. At necropsy, animals have been euthanized by cervical disloca tion following CO2 publicity. Tumors, livers, lungs, and brains had been removed and snap frozen or fixed in 10% formalin for long term evaluation.

Serial passaging of mammospheres inside the absence of TRAIL and/or FLIPi uncovered enrichment of MFUs in all cell cultures except those pre handled with each TRAIL and FLIPi. MFU enrichment is indicative of stem cell self renewal resulting from symmetric cell division. The comprehensive reduction of mammospheres from TRAIL/FLIPi handled cultures in subsequent passages suggests that the couple of surviving cancer initiating cells from 18 hours com bined treatment method had been severely compromised and not able to undergo additional symmetric cell divisions. Precisely the same effects have been also observed applying an different c FLIP siRNA target sequence. The ablation of practical MFUs represents a preferen tial sensitization of bCSCs to TRAIL compared towards the rest of your tumour cell population. This was confirmed by flow cytometry utilizing the marker ALDH1 which has previously been shown to enrich HER2 beneficial breast cancer cell populations for tumour initiating cells.
The HER2 good cell lines BT474 and SKBR3 had been subjected to TRAIL/FLIPi or control siRNA for 18 hrs and only the surviving adherent cells stained for ALDH1 exercise. The two cell lines exhibited signif icant reductions in the relative proportion of ALDH1 posi tive cells from the surviving populations following combined treatment method. In order selleck inhibitor to address which c FLIP isoform was responsi ble for the ablation from the self renewing action of your can cer stem cell population, siRNA sequences specific for cFLIP short and c FLIP lengthy transcripts have been utilised prior to mammosphere assay. Silencing of c FLIP long, but not c FLIP quick, mimicked the cytotoxic results of international c FLIP suppression in cancer stem cells, which confirmed an earlier observation of c FLIP lengthy mediated survival in MCF 7 cells. Suppression of c FLIP isoforms also sensitized cancer stem cells to sub toxic amounts of TRAIL.
TRAIL concentrations have been diminished from twenty ng/ml to one ng/ml, ranges that failed to activate a cell death response from the TRAIL delicate MDA MB 231 cell line, and mammosphere cultures were carried out as described over. TRAIL addition alone had diminished results on mammosphere integrity, nonetheless mixed treatment abrogated MFUs in BT474, SKBR3 and MDA MB 231 cell cultures, as previously observed selleck chemical with larger concentrations of TRAIL. The poorest responding cells to combined therapy, MCF seven, designed self renewing MFUs at very very low frequency in lowered TRAIL conditions. The mammosphere formation assay mainly exams the cells capability to resist anoikis, that’s a essential property of tumour initiating cells. As c FLIP has pre viously been reported to become an inhibitor of anoikis in other tumour cell sorts we wished to test regardless of whether the MFU sensitization to TRAIL was dependent on the added stresses imparted through the non adherent conditions.

Trastuzumab, a recombinant humanized monoclonal antibody directed towards an extracellular area of HER2, was the very first HER2 targeted treatment authorized from the United states of america Foods and Drug Administration for your treatment method of HER2 overexpressing MBC. Moreover, trastuzumab with adjuvant chemotherapy drastically enhanced disorder free and general survival charges in sufferers with early stage HER2 overexpressing breast cancer. Trastuzumab, mechanisms of antitumor results The mechanisms by which trastuzumab induces regression of HER2 overexpressing tumors are still getting elucidated, but numerous molecular and cellular results are actually reported while in the literature. Trastuzumab decreases signaling mediated by HER2 through the phosphatidylinositol 3 kinase and mitogen activated protein kinase cascades. Reduced downstream signaling by way of these pathways induces the cyclin dependent kinase inhibitor p27kip1, which promotes cell cycle arrest and apoptosis.
Trastuzumab quickly dissociates the non receptor tyrosine kinase Src from HER2, lowering Src action this kind of that the phosphatase and tensin homolog deleted on chromosome ten is dephosphorylated and translocated on the plasma membrane wherever it really is energetic. The PI3K down stream effectors Akt and mammalian target of rapamycin are then inhibited. The efficacy of trastuzumab can also rely on its pifithrin alpha capacity to induce an immune response. HER2 targeted antibodies, together with trastuzumab, had been shown to promote apoptosis in many breast cancer cell lines by way of antibody dependent cellular cytotoxicity. Importantly, mice that have been null to the Fc gamma receptor expressed on organic killer cells, that are vital for ADCC, misplaced considerably in the antitumor impact of trastuzumab, with only 29% tumor development inhibition observed versus 96% in control mice expressing the Fc gamma receptor and with intact all-natural killer cell perform.
Therefore, an lively immune response to trastuzumab might be partially accountable for cytotoxic exercise. Additionally, a larger in situ infiltration of leukocytes and ADCC action have been observed in sufferers achieving finish or partial remission after receiving preoperative trastuzumab relative to those who did not reply to this routine. Since patients with sophisticated MBC are BMS599626 immunosuppressed, it can be hard to enjoy the magnitude from the contribution of ADCC to trastuzumab mediated tumor inhibition. Extra in depth in vivo studies are necessary to grasp precisely how crucial the contribution of ADCC should be to mediating the response to trastuzumab and irrespective of whether other targeted antibodies made use of towards reliable tumors also depend on immune modulation to realize response. Trastuzumab has also been shown to inhibit angiogenesis, leading to decreased microvessel density in vivo and decreased endothelial cell migration in vitro.

IL 21 concentration was determined implementing a validated ELISA, the reduce limit of quantification of this assay was 0. 112 ng/mL. Sorafenib concentration selleck was established using liquid chromatography together with the tandem mass spectrometric de tection method. Cmax, AUC0 t, and t1/2, z were estimated using WinNonlin Expert v5. two. one soft ware. As a result of sparse sampling scheme, reported t1/2, z values must be interpreted with caution. Serum samples to find out soluble CD25 con centration applying a validated enzyme linked immuno sorbent assay were drawn at select time points during course one. Baseline serum and plasma VEGF ranges had been established using a validated immunoassay. The relationships of baseline VEGF and sCD25 ranges and that of transform in sCD25 levels to clinical efficacy endpoints had been explored.
Serum specimens were collected at decide on time points to display for IL 21 binding antibodies working with ELISA, samples containing IL 21 particular antibodies had been evaluated for neutralizing action by a cell based selleckchem Dabrafenib bioassay. Statistical evaluation Based to the binomial distribution, it had been determined that a sample dimension of 30 subjects inside the phase 2 portion on the study would supply around 95% probability of observing a related security occasion in 1 or far more subjects in case the true population incidence fee was 10% or higher. Offered the early phase of this study, formal assessments of energy for efficacy endpoints weren’t carried out. Kaplan Meier estimates for median PFS have been computed using the earliest assessment of progression treated as time of progression. SAS version 9. 1 was implemented to perform all analyses. The association in between baseline ranges of VEGF and sCD25, as well as sCD25 induction, with outcomes of clinical efficacy was explored using a series of Cox regression and logistic regression designs.
Introduction Metaplastic breast cancer is really a rare and histolog ically diverse subtype of breast carcinoma. It accounts for much less than 1% of all breast cancers. MBC was not officially recognized like a distinct histologic entity right up until 2000, and study on this disease approach continues to be limited as a result of its rarity along with the selection of tumor kinds included in this diagnosis. This class of breast malignancies encompasses tumors during which adenocarcinoma is noticed to co exist with an admixture of spindle cell, squamous, chondroid or bone forming neoplastic cells. Most metaplastic cancers are estrogen receptor, progesterone receptor and Her2 neu damaging and have a tendency to possess a worse prognosis than other triple detrimental breast cancers with fewer therapeutic choices. Within this evaluation, we discuss the histopathology and clinical features of MBC and their relevance to prognosis and systemic therapy alternatives likewise as long term instructions for investigate.

For those grafts surviving 10 years, nonetheless, the subsequent yearly failure fee has increased over time. Function of immunosuppression in long lasting outcomes The position of chronic immunosuppression in adverse patient outcomes has long been recognised. The contri bution to mortality from infection is famous. On top of that, the worsening of metabolic abnormalities such as diabetes and hyperlipidaemia and elevations in blood stress contribute to cardiovascular danger. Immunosup pression in blend with other oncogenic stimuli this kind of as viruses and ultraviolet radiation brings about signicant increases from the possibility of advancement of selected cancers this kind of as Kaposi sarcoma, lymphoma and nonmelanotic skin cancers. Dierent immunosuppressive drugs exhibit dierent proles of toxicity and may well inuence in dierent strategies the adverse outcomes viewed from the long run.
inhibitor Rucaparib Substantially attention is directed towards the contribution of chronic nephrotoxicity of calcineurin inhibitors to long run graft failure. A examine of protocol biopsies of renal allografts from 3 months to ten many years after transplantation by Nankivell and colleagues described the development of continual allograft nephropathy in excess of time. When alterations of CAN particularly, interstitial brosis and tubular atrophy developed within the rst yr after transplantation, these had been attributed to early events this kind of as rejection and acute tubular necrosis. Lesions this kind of as arteriolar hyalinosis and glomerulosclerosis developed later on and have been mentioned to indicate damage from cyclosporine exposure. Not too long ago, doubt has been cast within the relative contri bution of CNI nephrotoxicity to long-term graft failure relative to other leads to of graft harm this kind of as T cell mediated and antibody mediated rejection, polyoma virus infection and recurrent glomerulonephritis.
In the North American research of biopsies of failing grafts, the dominant TG100115 histological lesion noticed at five years was persistent antibody mediated rejection. Nevertheless, a lot of the biopsies were performed in response to deteriorating renal perform or other clinical occasions at a median time of 17 months soon after transplantation. This study therefore diers from the study by Nankivell and colleagues, where the biopsies have been carried out at protocol derived time factors to 10 years. There are actually very few studies where the challenging endpoints of graft loss or patient death give an indication with the adverse eects of long-term CNI therapy. One of these research will be the Australian Multicentre Cyclosporine Renal Transplant Study, which has lately published benefits following 20 many years of comply with up. Patients randomised to ongoing cyclosporine A had an inferior end result in contrast with patients who acquired CsA for 3 months in advance of converting to azathioprine and corticosteroid. This review for that reason supports that CNI exposure contri butes to deteriorating renal function and graft loss inside the really long run following renal transplantation.

Thus, by identifying the modifications from the luciferase shRNA versus untransduced experi ments and subtracting individuals, the subsequent pathway analyses could focus on pathways that were exclusively targeted by knockdown from the ASD related genes and never determine artifacts of your transduction. We next analyzed the gene lists from the shRNA experiments by two pathway analysis approaches to obtain diverse perspectives to the information. Essentially the most prominent pathways uncovered by analysis with NextBio were a number of pathways related to neurologic signaling and function. Secondarily, NextBio indicated that various pathways concerned common cellular metabolic process and growth had been also impacted.
One prominent pathway, the Peroxisome proliferator activated receptor gamma coactivator 1 A pathway, is based over the MSigDBs version of BioCartas pathway and has Mef2A and various calcium dependent kinases, which show gene expression modifications in all shRNA experiments. A single aspect on the NextBio analysis is the fact that directionality of transform is reported. Nearly all the pathways selleck chemicals Cediranib are downregulated with ASD shRNA knockdown, suggesting the genes we chose for this work are needed for your expression of these pathways and therefore their activity. Pathway evaluation with GenSensor also recognized several pathways relevant to neuronal signaling and perform. As using the NextBio evaluation, numerous development and metabolism pathways have been also impacted. For the duration of an examination from the individual pathways recognized through the two pathway evaluation techniques, we noted a recurring involvement from the mitogen activated protein kinase kinase /ERK signaling pathway.
These effects would happen both directly via a kinase signaling cascade or through cAMP. To further investigate this potential commonality, we employed CRE evaluation to identify possible underlying mechanisms in the shRNA datasets. As opposed to pathway analysis, which identifies pathways TW-37 solubility with altered gene expression, CRE examination predicts likely mechanisms behind gene modifications based within the concordance from the number of genes that alter expression, along with the directionality of that alter. The results of CRE examination are interlinked hypotheses of probable driving mechanisms or experimental treatment options that exhibit related gene improvements. It really is exciting that three of your eight most conserved hypotheses have a biological function suggestive of growth and/or immune function, suggesting similar driving mechanisms.
Likewise, you’ll find very conserved hypotheses concerned with neurogenesis, synaptic action and differentiation, as anticipated, while not mutually unique. Picking out the Mef2d experiment as being a representa tive from the 6 most conserved shRNA therapies, the major ranking clusters could be connected like a molecular inter action map with, cyclic AMP and ERK serving as dual hubs of your network immediately connecting 7 and eight related hypotheses respectively.

Following a protocol described by Irwin and Schoichet, molecules obtained through the Zinc database have been selected for comparison, given 95 99% very similar ity, together with the structures observed from the Kegg and PubChem databases. Additionally, se lected structures had been downloaded in mol2 and pdb formats for subsequent Docking Research and Molecular Dynamics. Docking studies The structures downloaded from your Kegg, Pubchem and Zinc databases were 1st checked in PyMOL one. 4 to assess the presence or absence of hydrogen, the stereochemistry of chiral carbons, substructure, superstructure, amount of rotatable bonds, amount of rings and number of hydrogen acceptors and donors. The ligand and receptor molecules were prepared in AutoDockTools 1. 5. six, All polar hydrogens had been additional to your receptor and Kollman United Atomic Fees had been computed.
For all ligands we additional polar hydrogens and computed the Gasteiger expenses. The grid definition, adjusted to the RPO lively website, was setup manually by fol lowing the recommendations from the system guide, The structures on the lig pop over to this website and and receptor were then saved in pqbqt format to be utilized for docking calculations. AutoDock Vina was used to perform Docking Scoring for every ligand receptor complicated, In advance of operating each Docking calculation, a configuration file was created with information about grid size and coordinates and indicating the ligand and receptor files. The reports for each calculation had been analyzed to obtain affinity vitality values for each ligand conformation in its respective complex. In addition, we employed PyMOL 1.
4 to verify the amount of hydrogen bonds and non covalent interactions be tween every ligand conformation and also the catalytic residues of RPO which have been concerned within the recognition and polymerization mechanisms. So as to optimize the preference of an ideal complicated we selected only one ligand that match greatest within the RPO active site, MK2206 thinking about all stereochemical elements previously evaluated plus the totally free vitality effects. Molecular dynamics of complicated On this study we utilized the MM PBSA protocol to determine affinity and stability from the ligand receptor RPO complicated interaction, using the package Amber twelve, At first, we utilized the Antechamber plan to make the ff99 force discipline identify the kinds of atom in each ligand and receptor and stay away from errors throughout the calculations.
tLEaP was utilized to neutralize charges and also the RPO ligand complicated was immersed in a rectangular box of TIP3P water molecules. Following the protocol we used Sander to carry out a Molecular Dynamics equilibrium, restricted to a area in the protein that incorporates the active web page, in accordance towards the following pa rameters. 1000 cycles of steepest descent and one thousand cycles of conjugate gradient minimization, heating MD for 200 picoseconds, density equilibrium for 200 ps, followed by Equilibrium Dynamics for 600 ps at continuous stress and 300 K.