Beta Elemene induces cytotoxic effects in FLT3 ITD-mutated acute myeloid leukemia by modulating apoptosis.

OBJECTIVE: ß-Elemene, a sesquiterpene with a broad anti-cancer spectrum, is particularly effective against drug-resistant and complex tumors. It can also be efficient against FLT3-expressed acute myeloid leukemia. This research aims to determine whether ß-Elemene has cytotoxic effects on FLT3 ITD-mutated AML cells. MATERIALS AND METHODS: Cytotoxicity, cell morphology, mRNA analysis with apoptotic markers, and analysis of 43 distinct protein markers related to cell death, survival, and resistance were all performed to elucidate its mechanism. Additionally, in order to understand how ß-Elemene and FLT3 interact, molecular docking, molecular dynamics simulations, and computational ADME investigations were performed. RESULTS: ß-Elemene exhibited cytotoxic activity against FLT3-mutated MV4-11 and FLT3 wild-type THP-1 cells, with an IC50 of around 25 µg/ml. The molecular studies revealed that ß-Elemene inhibited cell proliferation by inducing p53, and the involvement of p21, p27, HTRA, and HSPs were also demonstrated. The interactive inhibition in proliferation was confirmed via molecular docking and dynamics analyses. ß-Elemene occupied the FLT3 enzymatic pocket with good stability at the FLT3 active site. CONCLUSIONS: We concluded from our observations that ß-Elemene causes cell death in ITD mutant AML cells, together with the effects of stress factors and inhibiting cell division.

The opioid peptide biphalin modulates human corneal epithelial wound healing in vitro.

PURPOSE: Analgesic drugs, including nonselective opioids and non-steroidal anti-inflammatory drugs, should be used with great precautions to relieve pain after trauma to the corneal epithelium because of their unfavorable effects on wound healing. Biphalin is a synthetic opioid peptide that has been demonstrated to possess a strong analgesic effect on rodents. The purpose of this study is to investigate the effects of biphalin on human corneal epithelial wound healing. METHODS: An immortalized human corneal epithelial cell (HCEC) culture was used to analyze the effects of biphalin on wound healing. The toxicity of biphalin at various concentrations was measured by the MTT assay. The effects of 1µM and 10µM biphalin on wound closure, cell migration and proliferation were tested in an in vitro scratch assay of HCECs. Naloxone, a non-selective competitive opioid receptor antagonist, was also used to inhibit the effects of biphalin in all experiments. RESULTS: Biphalin did not cause any toxic effect on HCECs at concentrations lower than 100µM at various incubation time points. Biphalin significantly increased wound healing at 1µM concentration in an in vitro scratch assay of HCECs (P<0.05). It also significantly increased migration of HCECs (P<0.01). There was no significant difference between the biphalin and control groups of HCECs in the Ki67 proliferation assay. CONCLUSION: Biphalin, which is a synthetic opioid peptide, promotes corneal epithelial wound healing by increasing cell migration. This role should be evaluated in further in vivo and clinical studies.

Active compounds extracted from extra virgin olive oil counteract mesothelial-to-mesenchymal transition of peritoneal mesothelium cells exposed to conventional peritoneal dialysate: in vitro and in vivo evidences.

During peritoneal dialysis (PD), peritoneal mesothelial cells undergo a transition from an epithelial phenotype to a mesenchymal phenotype that, together with the inflammatory process, promotes tissue fibrosis and a failure of peritoneal membrane function. To date, there is no definitive treatment for the progressive thickening and angiogenesis of the peritoneal membrane associated with PD. In this study we tested, in vitro and in vivo, the ability of active compounds extracted from extra virgin olive oil (AC-EVOO) to counteract the mesothelial-to-mesenchymal transition process (MMT) observed in mesothelial cells chronically exposed to the conventional peritoneal dialysate (DL). In particular, we used a cultivar from southern Italy known to have a high polyphenol content. Our results showed that, in mesothelial cells exposed to DL, the combined treatment with AC-EVOO prevented the genic and protein upregulation of key mesenchymal and inflammatory markers, as well as the MCs' migratory capacity. Concomitantly, we tested the antifibrotic efficacy of AC-EVOO in mesothelial cells obtained from effluents of patients undergoing PD, whose "fibroblast-like" phenotype was defined by flow-cytometry assay. We observed that in these cells AC-EVOO significantly mitigated, but did not reverse, the MMT process. In conclusion, our preliminary results suggest that AC-EVOO can interfere with critical factors in the process of differentiation, preventing myofibroblast formation, but once fibrosis has already progressed it is unable to promote the redifferentiation to the epithelial phenotype. Further studies are needed to establish whether AC-EVOO could represent a new therapeutic target to prevent peritoneal fibrosis.

Early cytomegalovirus-specific T-cell response and estimated glomerular filtration rate identify patients at high risk of infection after renal transplantation.

BACKGROUND: Assessing the risk of cytomegalovirus (CMV) viremia in kidney transplant recipients (KTR) may be helpful to indicate in which patient it is worth starting antiviral treatment during preemptive strategy. METHODS: In 40 CMV-seropositive KTR preemptively treated with ganciclovir, we used interferon (IFN)-γ ELISpot test to evaluate whether monitoring T cells directed against phosphoprotein (pp) 65 and immediate early (IE)-1 antigens could predict the onset of viremia. RESULTS: CMV viremia occurred in 24 patients (60%) within 120 days after transplantation. Non-viremic patients had higher anti-pp65, anti-IE-1 T cells, and estimated glomerular filtration rate (eGFR) in the first 90 days after transplantation. At logistic regression, anti-pp65, anti-IE-1 T cells, and eGFR measured at day 30 were significantly associated with CMV infection. Cutoff values of 15 spot-forming cells (SFCs)/200,000 peripheral blood mononuclear cells (PBMCs) for anti-IE, 40 SFCs/200,000 PBMCs for anti-pp65, and 46.6 mL/min/1.73 m(2) for eGFR, respectively, predicted the risk of CMV infection with high sensitivity and specificity (area under the receiver operating characteristic curve >0.75). Using a classification tree model, we identified as high-risk patients those showing anti-pp65 <42 SFCs/200,000 PBMCs and eGFR <62 mL/min/1.73 m(2) , as well as anti-pp65 ≥42 and anti-IE-1 <6.5 SFCs/200,000 PBMCs. CONCLUSION: Monitoring CMV-specific T-cell responses and eGFR in the first month post transplant can identify patients at high risk of CMV infection, for whom preemptive antiviral therapy is recommended.

Laparoscopic placement of "self-locating catheter": our experience and a review of literature.

Among the available devices for peritoneal dialysis, the Di Paolo self-locating catheter (SLC) represents a milestone using to its ability to ensure a permanent reliable means of access to the peritoneum. Our experience included 20 laparoscopic peritoneal catheter placements from 2008 to 2011. We performed the laparoscopic surgical technique using 3 trocars: 2 10 mm and 1 5 mm. The technique allows catheter introduction into the pouch of Douglas under direct vision. Among 20 treated patients, 1 died due to causes unrelated to peritoneal dialysis; 1 underwent transplantation, and 1 was switched to hemodialysis because of ultrafiltration failure. The complications included 2 catheter displacements, only 1 of them needing repositioning by open laparotomy, and 1 case of peritonitis. No infection in the subcutaneous tunnel or obstruction and malfunction occurred among our patients. The Di Paolo SLC is similar to Tenckhoff catheter but includes a small tungsten cylinder at the tip that engenders continuous gravity in the peritoneal cavity, producing a reduced risk of dislocation. In a large series of cases, Di Paolo et al. reported a 0.8% dislocation rate after SLC placement compared with 12% using Tenckhoff catheters. They also demonstrated a reduced risk of other complications, such as peritonitis, infection, obstruction, and failure. These data have been confirmed by other authors with smaller case series. Thus, introduction of the SLC and improved surgical techniques result in better efficiency of peritoneal dialysis.

GPE and GPE analogues as promising neuroprotective agents.

The tripeptide glycine-proline-glutamate (GPE) is the naturally cleaved N-terminal tripeptide of insulin-like growth factor-1 (IGF-1) in brain tissues by an acid protease. Although GPE does not bind to IGF-1 receptors and its mode of action is not clear, in vitro studies have demonstrated its ability to stimulate acetylcholine and dopamine release, as well as to protect neurones from diverse induced brain injures. More importantly, GPE has been shown to have potent neuroprotective effects in numerous animal models of hypoxic-ischemic brain injury and neurodegenerative diseases such as Parkinson's, Alzheimer's and Huntington's diseases. As a consequence, GPE was suggested to be a potential target for the rational design of neuroprotective agents. Unfortunately, the use of GPE as a therapeutic agent is limited because of its unfavorable biochemical and pharmacokinetic properties. This review will focus on structural modifications performed on the GPE molecule in order to obtain bioactive analogues with increased pharmacokinetic profile useful for the treatment of central nervous system (CNS) injures and neurodegenerative disorders.

2,5-diketopiperazines as neuroprotective agents.

2,5-diketopiperazines are the simplest cyclic peptides found in nature, commonly biosynthesized from amino acids by different organisms, and represent a promising class of biologically active natural products. Their peculiar heterocyclic structure confers high stability against the proteolysis and constitutes a structural requirement for the active intestinal absorption. Furthermore, the diketopiperazine-based motif is considered as a novel brain shuttle for the delivery of drugs with limited ability to cross the blood-brain barrier (BBB) and can be proposed as an ideal candidate for the rational development of new therapeutic agents. Although these cyclic peptides have been known since the beginning of the 20th century, only recently have they attracted substantial interest with respect to the wide spectrum of their biological properties, including antitumor, antiviral, antifungal, antibacterial and antihyperglycemic activities. In addition to these, the most challenging function of the diketopiperazine derivatives is related with their remarkable neuroprotective and nootropic activity. The aim of the present paper is to provide an overview of the two major classes of diketopiperazines, the TRH-related and the unsaturated derivatives both characterized by a significant ability to protect against neurotoxicity in several experimental models. The neuroprotective profile of these compounds suggests that they may have a future utility in the therapy of neuronal degeneration in vivo, potentially through several different mechanisms.

Thyroid status and kidney transplantation outcomes.

INTRODUCTION: It is known that end-stage renal disease patients can display abnormal thyroid gland function, which may cause autoimmune hypothyroidism or subclinical alterations. The impact of thyroid function on graft outcomes is not completely clear among renal transplant patients. The aim of this study was to evaluate thyroid function among a cohort of 136 consecutive renal recipients in correlation with clinical parameters of graft function. MATERIALS AND METHODS: We performed a cross-sectional study on 136 subjects including 84 males and 52 females of overall mean age of 49.71 ± 10.98 years who underwent renal transplantations between 2005 and 2009 and had a mean follow-up of 28.3 ± 15.7 months. All patients were treated with a calcineurin inhibitor, steroids, and mycophenolate mofetil. The exclusion criteria were age below 18 years, multiorgan transplantation, graft failure in the first 6 months, or presence of a thyroid neoplasm. We evaluated levels of serum FT3, FT4, and thyroid-stimulating hormone (TSH) in relation to the following parameters: body mass index (BMI), serum creatinine, estimated glomerular filtration rate estimated glomerular filtration rate (eGFR) by Modification of Diet in Renal Disease (MDRD) formula, proteinuria/24 hours, serum sodium, potassium, calcium, phosphorus, cholesterol, high-density lipoprotein, low-density lipoprotein, triglycerides, and hemoglobin (Hb). RESULTS: Only 6.4% of our transplant recipients were treated with levothyroxine sodium. The patients showed an average FT3 of 3.24 ± 0.5 mg/dL; average FT4 of 0.84 ± 0.1 mg/dL, and mean TSH of 1.29 ± 0.8 mg/dL. The study showed no relationship between thyroid hormones and age of the transplant, while there was a significant difference in FT3 levels between men and women. We also observed a significant correlation between FT3 and serum creatinine, eGFR, serum sodium, BMI, and Hb; whereas there was no correlation with other variables. The correlations between FT4 and TSH and all examined variables were not significant. CONCLUSIONS: The interactions between the thyroid and the kidney have been incompletely studied among patients with renal transplants. Our data showed that the presence of low serum FT3 levels correlated with worse graft function, anemia, BMI, and serum sodium. Thus low FT3 levels could be predictive of graft function, especially in the 5 years posttransplantation.

Hybrid alpha/beta-peptides: for-Met-Leu-Phe-OMe analogues containing geminally disubstituted beta2,2- and beta 3,3-amino acids at the central position.

The two fMLF-OMe analogues For-Met-beta(3)hAc(6)c-Phe-OMe (6) and For-Met-beta(2)hAc(6)c-Phe-OMe (12) and their corresponding N-Boc derivatives 5 and 11 have been synthesized and their biological activity towards human neutrophils evaluated. The N-formyl peptides 6 and 12 exhibit good activity as chemoattractans and 12 is highly active in superoxide anion production. The preferred solution conformation of the two N-formyl derivatives has been discussed.

Carboplatin sustained delivery system using injectable microspheres.

A controlled carboplatin delivery system using biodegradable polymer has been used in this study. The purpose was to evaluate the local and systemic effects of injectable, biodegradable microspheres containing carboplatin when injected as aqueous suspension subcutaneously in rats. Biocompatibility and toxicity of empty microspheres and microspheres loaded with carboplatin were evaluated by clinical and histological examination. The diffusion of carboplatin in tissues and time of drug release were evaluated by platinum determination in plasma and tissues over the time. The results of the study suggest that microspheres provide a sustained slow release of carboplatin and that multiple inoculations of microspheres containing drug and no evidence of local or systemic toxicity is found. This device may be useful in the treatment of solid tumours.

On-line biofilm monitoring by "BIOX" electrochemical probe.

The innovative electrochemical monitoring probe (BIOX) recently developed to improve the antifouling treatments of cooling systems in industrial plants is presented. On the basis of the good results obtained from applications on marine sites, some research has been stated to validate this technique in biofilm growth and prevention of microbial corrosion in fresh and drinking waters.

Isopeptide bonds in chemotactic tripeptides. Synthesis and activity of lysine-containing fMLF analogs.

In order to explore the properties of chemotactic N-formylpeptides containing isopeptide bonds within their backbones, a group of lysine-containing analogs of the prototypical chemotactic tripeptide N-formylmethionyl-leucyl-phenylalanine (fMLF) was synthesized. The new analogs were designed by adding to the HCO-Met or Boc-Met residue a dipeptide fragment made up of Lys and Phe residues joined through Lys N alpha or N epsilon bonds, in all possible combinations. Thus, the following six pairs of tripeptides were synthesized and examined for their bioactivity: RCO-Met-Lys(Z)-Phe-OMe (2a, b), RCO-Met-Lys(Z-Phe)-OMe (3a, b), Z-Lys(RCO-Met)-Phe-OMe (4a, b), Z-Phe-Lys(RCO-Met)-OMe (5a, b), RCO-Met-Phe-Lys(Z)-OMe (6a, b) and Z-Lys(RCO-Met-Phe)-OMe (7a, b), with R=OC(CH3)(3 )and R=H for compounds a and b, respectively. All the new models were characterized fully and their activity (chemotaxis, superoxide anion production and lysozyme release) on human neutrophils determined as agonists (compounds b) and antagonists (compounds a). All N-formyl derivatives 2b-7b are less potent than fMLF-OMe as chemoattractants, but compound 7b exhibits selective activity as superoxide anion producer. Derivatives 2a-7a do not show antagonistic activity towards fMLF induced chemotaxis and O(2)(-) production, however, all these compounds except 4a antagonize lysozyme release by 60%.

Gorham's disease: a literature review and case reports.

Gorham's disease (disappearing bone disease, massive osteolysis, idiopathic osteolysis, essential osteolysis, progressive atrophy of bone, spontaneous absorption of bone, phantom bone, hemangiomatosis/lymphangiomatosis of bone, progressive osteolysis) is an extremely rare occurrence. There are fewer than 150 reported cases in the literature. This disorder can be characterized by spontaneous or posttraumatic progressive resorption of bone. The etiology is still very speculative, the prognosis unpredictable, and any effective therapy still unknown. This paper presents a review of the literature and two case reports of suspected Gorham's disease of the bones of the foot.

A statistical analysis on the reliability of the proximal articular set angle.

The authors investigated the statistical relationship and significance between the transverse and sagittal plane proximal articular set angles both radiographically and intraoperatively. The analysis revealed a highly significant difference between the means of the respective measurements. Although the radiographic and intraoperative findings were found to be related for the transverse plane proximal articular set angle, the transverse plane PASA was approximately 7 degrees greater when measured intraoperatively. The concept of a sagittal plane PASA was also introduced. The significance of accurate measurements in the surgical correction of hallux valgus was also examined.