bluuebunny/crossref_metadata_2025 huggingface dataset:
Review of Quality Deficiencies Found in Active Pharmaceutical Ingredient Master Files Submitted to the WHO Prequalification of Medicines Programme
Authors: Ortega Diego Isabel , Fake Antony , Stahl Matthias , Rägo Lembit
Published: March 2016

Purpose. The aim of this work was to determine the number and type of active pharmaceutical ingredient (API) quality deficiencies in API Master Files (APIMFs) as submitted to the World Health Organization (WHO) Prequalification of Medicines Programme (PQP). Methods. We conducted a retrospective review of API quality deficiencies identified following the assessment of new APIMFs for non-sterile APIs during a 6-year period from 1 January 2007 to 31 December 2012. All deficiencies were collected, classified and quantified according to the Common Technical Document (CTD) sections and subsections and as groups of commonly raised questions. Results. There were 5446 deficiencies collected from 159 APIMF deficiency letters by CTD section, by selected CTD subsections and by selected CTD subsections and year. More than 50% of the total number of deficiencies related to the manufacturing sections of the CTD, followed by deficiencies concerning the impurities, the API specification and the stability sections of the CTD.  A pattern of API deficiencies across the different CTD subsections and over time was identified. Conclusions. The most frequent critical deficiencies were related to how the specific manufacturing process and the key materials used, in particular the API starting material, impact the API impurities content. The number and pattern of APIMF deficiencies did not change over time. The results are compared to the findings in similar studies as reported by the United States Food and Drug Administration (USFDA), the European Directorate for the Quality of Medicines (EDQM) and the European Medicines Agency (EMA) and similarities and differences are discussed. Our findings highlight the need for greater guidance and technical assistance for API manufacturers submitting APIMFs to the PQP. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.
Using novobiocin as a specific inhibitor of breast cancer resistant protein to assess the role of transporter in the absorption and disposition of topotecan
Authors: Su Yaming , Hu Peidi , Lee Sung-Hack , Sinko Patrick J
Published: March 2016

Purpose. To investigate the role of intestinal breast cancer resistant protein (BCRP) in the absorption and disposition of topotecan (TPT) using novobiocin (NOV) as a specific inhibitor. Methods. Transporter inhibition specificity of NOV was assessed in cells overexpressing BCRP or Pgp. Sprague-Dawley rats were orally or intravenously dosed with TPT (2 and 1 mg/kg for p.o. and i.v., respectively) with or without oral co-administration of NOV (50 mg/kg). Pharmacokinetic parameters of TPT were obtained by noncompartmental analysis. To assess the role of BCRP in TPT intestinal permeation, rat ileal segment was perfused with 10 uM TPT in the presence or absence of NOV (500 uM), TPT permeability was calculated based on drug appearance in mesenteric blood. To assess the role of BCRP in TPT intestinal secretion, rat ileal segment was perfused with saline in the presence or absence of NOV (500 uM), while TPT was i.v. infused into rat. Intestinal secretion of TPT was calculated based on drug appearance in the perfusate. Results. NOV significantly inhibited efflux activity of BCRP, but not Pgp. Coadministration of NOV markedly increased oral TPT AUC(0–720) and Cmax by 3- and 4.5-fold, respectively, and decreased systemic clearance of i.v. injected TPT (from 44.40±7.28 without NOV to 29.44±1.99 ml/min/kg with NOV). The inclusion of NOV in perfusate significantly increased TPT permeability from 0.81±0.30 x10(-6) to 1.26±0.12 x10(-6) cm/s, while, the intestinal secretion of TPT was reduced by ~50% when NOV was included in perfusate. Conclusions. The present study establishes in vitro and in vivo inhibition potency and specificity of NOV on BCRP and provides direct evidence that intestinal BCRP plays an important role in limiting the oral absorption and influencing the systemic elimination of TPT.
In vitro Anti-Inflammatory Effects of Hyaluronic Acid in Ethanol-Induced Damage in Skin Cells
Authors: Neuman Manuela G , Nanau Radu M , Oruña Loida , Coto Gabriel
Published: March 2016

ABSTRACT - Ethyl alcohol (ethanol) is commonly applied in cosmetic and pharmaceutical preparations, as well as disinfectant for chronic wounds. Objective: The present study aimed to appraise physiological levels of ethanol-induced damage in skin cells in vitro and the possible repair by hyaluronic acid (HA). In addition, we aimed to establish cytokine-chemokine networks in the cellular media and the modulation of cytokines such as tumor necrosis factor-alpha (TNF-B), interferon-alpha (IFN-α), transforming growth factor-beta (TGF-B), interleukins (IL) such as IL1-B and IL-6, as well as matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP). Design and Methods: We treated human A431 epidermoid skin cells and mouse fibroblasts with ethanol at a concentration of 100 mM over 24 hours (h). A separate experiment looked at the effects of 2 consecutive treatments with 100 mM ethanol for 24 h each. HA obtained from umbilical cord excision was used at two concentration levels (2% and 4%) to determine its efficacy in the treatment. We measured cytotoxicity and cytokine networks in the media. Results: Treatment of cells with 100 mM ethanol increased cytotoxicity, as well as the release of pro-inflammatory cytokines into the culture medium. Conclusions: Ethanol may induce cytotoxicity in skin cells by enhancing the effects of pro-inflammatory cytokines. HA reduced the amount of pro-inflammatory cytokines released into the media both in human A431 epidermoid skin cells and in mouse fibroblasts.

This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.
Cationic Solid Lipid Nanoparticles Loaded by Cystein Proteinase Genes as a Novel anti-Leishmaniasis DNA Vaccine Delivery System: Characterization and in vitro Evaluations
Authors: Doroud Delaram , Vatanara Alireza , Zahedifard Farnaz , Gholami Elham , Vahabpour Rouhollah , Rouholamini Najafabadi Abdolhossein , Rafati Sima
Published: March 2016

Purpose: Leishmaniasis is a major health problem in many tropical and sub-tropical countries and development of a safe and easily-available vaccine has high priority. Although several antigens potentially capable of inducing protective immunity have been studied, in the absence of pharmaceutical industry interest they have remained as fine publications only. Amongst them, Cathepsin L-like cysteine proteinases (CPs) have received considerable attention and type I and II CPs have been used in a heterologous prime-boost vaccination regime for experimental visceral leishmaniasis in dogs. Due to the promising results of the mentioned vaccination regime, we aimed to evaluate cationic solid lipid nanoparticles (cSLNs) for in vitro delivery of cpa, cpb and cpbCTE intended to be used as a cocktail DNA vaccine in our forthcoming studies.

Methods: cSLNs were formulated of cetyl palmitate, cholesterol, DOTAP and Tween 80 via melt emulsification method followed by high shear homogenization. Different formulations were prepared by anchoring pDNAs on the surface of cSLNs via charge interaction. The formulations were characterized according to their size and zeta potential as well as pDNA integrity and stability against DNase I treatment. Lipoplexes' cytotoxicity was investigated on COS-7 cells by MTT test. The effect of the DOTAP:pDNA ratio on protection ability and cytotoxicity was also studied. In vitro transfection efficiency was qualified by florescent microscopy and quantified using flow cytometry technique.

Results: cSLN-pDNA complexes were formulated with suitable size and zeta potential. Efficiency/cytotoxicity ratio of cSLN-pDNAs formulations was comparable to linear PEI-25KD-pDNAs polyplexes while exhibiting significantly lower cytotoxicity.

Conclusion: Tested formulations were able to deliver immunogenic CP genes efficiently. This data proves the ability of this system as a promising DNA vaccine carrier for leishmaniasis to cover the main drawback of naked pDNA delivery that is rapid elimination from the circulation.
Design and Pharmaceutical Evaluation of a Nano-Enabled Crosslinked Multipolymeric Scaffold for Prolonged Intracranial Release of Zidovudine
Authors: Harilall Sheri-Lee , Choonara Yahya E , Modi Girish , Tomar Lomas K , Tyagi Charu , Kumar Pradeep , Du Toit Lisa C , Iyuke Sunny E , Danckwerts Michael P , Pillay Viness
Published: March 2016

Purpose. Nanomedicine explores and allows for the development of drug delivery devices with superior drug uptake, controlled release and fewer drug side-effects. This study explored the use of nanosystems to formulate an implantable drug delivery device capable of sustained zidovudine release over a prolonged period. Methods. Pectin and alginate nanoparticles were prepared by applying a salting out and controlled gelification approach, respectively. The nanoparticles were characterized by attenuated total reflectance-fourier transform infrared spectroscopy (ATR-FTIR), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and dynamic light scattering (DLS) and were further evaluated for zidovudine (AZT) entrapment efficiency. Multipolymeric scaffolds were prepared by crosslinking carboxymethyl cellulose, polyethylene oxide and epsilon caprolactone for entrapment of zidovudine-loaded alginate nanoparticles to impart enhanced controlled release of zidovudine over the time period. Swelling and textural analysis were conducted on the scaffolds. Prepared scaffolds were treated with hydrochloric acid (HCl) to reduce the swelling of matrix in the hydrated environment thereby further controlling the drug release. Drug release studies in phosphate buffered saline (pH 7.4, 37°C) were undertaken on both zidovudine-loaded nanoparticles and native scaffolds containing alginate nanoparticles. Results. A higher AZT entrapment efficiency was observed in alginate nanoparticles. Biphasic release was observed with both nanoparticle formulations, exhibiting an initial burst release of drug within hours of exposure to PBS, followed by a constant release rate of AZT over the remaining 30 days of nanoparticle analysis. Exposure of the scaffolds to HCl served to reduce the drug release rate from the entrapped alginate nanoparticles and extended the AZT release up to 30 days. Conclusions. The crosslinked multipolymeric scaffold loaded with alginate nanoparticles and treated with 1% HCl showed the potential for prolonged delivery of zidovudine over a period of 30 days and therefore may be a potential candidate for use as an implantable device in treating Aids Dementia Complex.

This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.
Applications of β-limit Dextrin as a Matrix Forming Excipient for Fast Disintegrating Buccal Dosage Formats
Authors: Qi Xin , Tester Richard , Liu Yu , Mullin Margaret
Published: March 2016

ABSTRACT - Purpose. To compare the properties of buccal delivery matrices (wafers) made with dextrin, β-limit dextrin and pre-gelatinised starch. Methods. The constituent α-glucans were tested for their mucoadhesive properties in solution plus their content of crystalline material (differential scanning calorimetry, DSC). Wafers were made by lyophilisation of aqueous solutions/dispersions of the α-glucans. Physical properties of the wafers were evaluated using texture analysis, dissolution coupled to photography and scanning electron microscopy (SEM). Results. The results highlighted how the β-limit dextrins chemical and physical properties were ideally suited for the production of buccal delivery wafers. Dissolution testing confirmed the excellent hydration profile of the β-limit dextrin (within wafers) with time. Using SEM it was evident that the homogeneous “bee-hive” like structure of the β-limit dextrin wafers, unlike the other α-glucans, provided a rapidly hydratable strong porous matrix. Conclusions. The β-limit dextrin α-glucan makes a superb (lyophilised) mucoadhesive delivery structure for the delivery of active agents to the buccal mucosa.

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Population Pharmacokinetic/Pharmacodynamic Analysis of the DPP-4 Inhibitor Linagliptin in Japanese Patients with Type 2 Diabetes Mellitus
Authors: Tadayasu Yusuke , Sarashina Akiko , Tsuda Yasuhiro , Tatami Shinji , Friedrich Christian , Retlich Silke , Staab Alexander , Takano Mikihisa
Published: March 2016

[Objectives] Linagliptin is a novel, highly selective and long acting DPP-4 inhibitor for the treatment of type 2 diabetes mellitus (T2DM). Linagliptin exhibits non-linear pharmacokinetics (PK) due to saturable binding to plasma and tissue DPP-4. The aim of this study was to characterize the PK and PK/DPP-4 inhibition relationship of linagliptin in Japanese patients with T2DM using a population PK/DPP-4 model and to support the rationale for the therapeutic dose in Japanese patients by simulation. [Methods] Linagliptin plasma concentration and DPP-4 inhibition measurements from a placebo-controlled, parallel group multiple (28 days) dose trial that included 36 T2DM patients (18 patients each in 2.5 mg and 10 mg dose group) were used for analysis. Modeling was performed using FOCE INTERACTION estimation method implemented in NONMEM V. The linagliptin plasma concentration- and DPP-4 inhibition- time profiles were simulated for Japanese patients receiving 5 mg linagliptin once daily by the model established. [Results] Nonlinear PK of linagliptin in T2DM patients were well described by a 2-compartment model assuming concentration-dependent binding to DPP-4 in the central and peripheral compartment. Plasma DPP-4 inhibition was integrated in the model by relating the model-predicted DPP-4 occupancy with linagliptin linearly to DPP-4 inhibition. The simulation predicted that for the 5 mg dose group the trough DPP-4 inhibition at steady-state was 84.2%, which is higher than the target inhibition (≥80%) for an effective dose of DPP-4 inhibitor. In 2.5 mg dose group, steady-state DPP-4 inhibition of >80% was not maintained over 24 hours (observed and simulated). [Conclusions] The nonlinear PK of linagliptin and its plasma DPP-4 inhibition in patients were well characterized by a target-mediated drug disposition model relating DPP-4 occupancy with linagliptin to DPP-4 inhibition. Simulations of plasma DPP-4 inhibition suggest that 5 mg linagliptin once daily is an appropriate therapeutic dose for Japanese patients with T2DM.This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.
Bioreduction of Idarubicin and Formation of ROS Responsible for DNA Cleavage by NADPH-Cytochrome P450 Reductase and its Potential Role in the Antitumor Effect
Authors: Çelik Haydar , Arinç Emel
Published: March 2016

PURPOSE. Idarubicin is a clinically effective synthetic anthracycline analog used in the treatment of several human neoplasms. Anthracyclines have the potential to undergo bioactivation by flavoenzymes to free radicals and thus exert their cytotoxic actions. In this study, our main objective was to investigate the possible involvement of NADPH-cytochrome P450 reductase in the bioreductive activation of idarubicin to DNA-damaging species. METHODS. A pBR322 plasmid DNA damage assay was used as a sensitive method for detecting strand breaks in DNA exposed to idarubicin in the presence of P450 reductase and cofactor NADPH under various incubation conditions. In addition, the rates of idarubicin reduction by P450 reductases purified from phenobarbital-treated rabbit liver, beef liver and sheep lung microsomes were determined by measuring NADPH oxidation at 340 nm. RESULTS. The plasmid DNA experiments demonstrated that idarubicin could undergo bioreduction by P450 reductase with the resulting formation of DNA strand breaks. The antioxidant enzymes SOD and catalase, and hydroxyl radical scavengers, DMSO and thiourea, afforded significant levels of protection against idarubicin-induced DNA strand breaks. These findings suggested that DNA damage by idarubicin occurs through a mechanism which involves its redox cycling with P450 reductase to generate reactive oxygen species (ROS). The extent of DNA damage by idarubicin was found to increase with increasing concentrations of drug or enzyme as well as with increasing incubation time. The capacity of idarubicin to induce DNA damage under above incubation conditions was compared with that of a model compound, mitomycin C. Finally, enzyme assays carried out with purified P450 reductases revealed that idarubicin exhibited about two-fold higher rate of reduction than mitomycin C. CONCLUSION. Our findings implicated bioreduction of idarubicin by P450 reductase and subsequent redox cycling under aerobic conditions as being one mode of idarubicin action potentially contributing to its antitumor effect.
The metabolism of amiodarone by various CYP isoenzymes of human and rat, and the inhibitory influence of ketoconazole
Authors: Elsherbiny Marwa E. , El-Kadi Ayman O.S. , Brocks Dion R.
Published: March 2016

PURPOSE. To evaluate the metabolism of amiodarone (AM) to desethylamiodarone (DEA) by selected human and rat cytochrome P450, and the inhibitory effect of ketoconazole (KTZ). METHODS. Some important CYP isoenzymes (rat CYP1A1, 1A2, 2C6, 2C11, 2D1, 2D2, and 3A1 and human CYP1A1, 1A2, 2D6 and 3A4) were spiked with various concentrations of AM to determine the relative kinetic parameters for formation of DEA in the presence and absence of various concentrations of KTZ. RESULTS. The formation of DEA was observed when AM was exposed to each of the CYP tested, although the rates were varied. Human CYP1A1 followed by 3A4 had the highest intrinsic clearance (CLint) for DEA formation whereas in rat, CYP2D1 followed by CYP2C11 had the highest CLint. Human and rat CYP1A2 seemed to have the lowest CLint. At high concentrations of AM and KTZ, near those expected in vivo, significant inhibition of all isoforms except for rat CYP1A2 was observed. At lower concentration ranges of both drugs, the inhibitory constant was determined. At these levels, KTZ was found to potently inhibit human CYP1A1 and 3A4 and rat 2D2 and 1A1. CONCLUSION. Human CYP1A1 and 3A4 and rat CYP2D1 and 2C11 were most efficient in converting AM to DEA. For DEA formation, the in vivo administration of KTZ could inhibit other CYP isoforms besides CYP3A in human and rat.
Random Allocation in Controlled Clinical Trials: A Review
Authors: Egbewale Bolaji Emmanuel
Published: March 2016

PURPOSE: An allocation strategy that allows for chance placement of participants to study groups is crucial to the experimental nature of randomised controlled trials. Following decades of the discovery of randomisation considerable erroneous opinion and misrepresentations of its concept both in principle and practice still exists. In some circles, opinions are also divided on the strength and weaknesses of each of the random allocation strategies. This review provides an update on various random allocation techniques so as to correct existing misconceptions on this all important procedure. METHODS: This is a review of literatures published in the Pubmed database on concepts of common allocation techniques used in controlled clinical trials.  RESULTS: Allocation methods that use; case record number, date of birth, date of presentation, haphazard or alternating assignment are non-random allocation techniques and should not be confused as random methods. Four main random allocation techniques were identified. Minimisation procedure though not fully a random technique, however, proffers solution to the limitations of stratification at balancing for multiple prognostic factors, as the procedure makes treatment groups similar in several important features even in small sample trials. CONCLUSIONS: Even though generation of allocation sequence by simple randomisation procedure is easily facilitated, a major drawback of the technique is that treatment groups can by chance end up being dissimilar both in size and composition of prognostic factors. More complex allocation techniques that yield more comparable treatment groups also have certain drawbacks. However, it is important that whichever allocation technique is employed, unpredictability of random assignment should not be compromised. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.
Statins Against Drug-Induced Nephrotoxicity
Authors: Dashti-Khavidaki Simin , Moghaddas Azadeh , Heydari Behrooz , Khalili Hossein , Lessan-Pezeshki Mahboob , Lessan-Pezeshki Mahbooh
Published: March 2016

Drug-induced nephrotoxicity (DIN) accounts for up to 60% of hospital acquired acute kidney injury with considerable morbidity and mortality. Several efforts have been made to reduce drug-induced nephrotoxicity; however, DIN remains a matter of concern. Statins with their antioxidant, anti-inflammatory and anti-apoptotic effects may have the potential to protect kidney against DIN. The present review evaluated all of the available in vitro and in vivo studies that examined the use of statins as renoprotective agents against nephrotoxic drugs. Materials for this review were obtained by searching Medline, PubMed, Scopus, Cochrane central register of controlled trials, and Cochrane database of systematic reviews. Key words used as search terms included “statin”, “3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors, “HMG-CoA reductase inhibitors”, “nephroprotective”, “renoprotective”, “drug-induced renal diseases”, “drug-induced nephrotoxicity”, “drug-induced renal toxicity”, “drug-induced nephropathy”, “drug-induced renal side effects”, and “contrast-induced nephropathy”. This search was performed without time limitation. Only English language articles were included in this review. This review concluded that chronic statin user may be less prone to contrast-induced nephropathy (CIN) compared with statin non-users. Short-term high dose statin administration may also reduce the incidence of CIN in statin naïve patients. This renoprotective effect of statins against CIN is seen in low risk patients with normal kidney function or mild kidney dysfunction, but probably not in patients with moderate to severe renal dysfunction.  Based on available animal data, statins may protect kidney against gentamicin-, cisplatin- and cyclosporine-induced nephrotoxicity, however, theses animal results have not yet been confirmed by human data. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.
Effects of pioglitazone on erectile dysfunction in sildenafil poor-responders: a randomized, controlled study
Authors: Gholamine Babak , Motevallian Manijeh , Shafiei Massoumeh , Mahmoudian Massoud
Published: March 2016

ABSTRACT

Purpose.To assess the effects of pioglitazone on sildenafil responsiveness in men with erectile dysfunction (ED) and a history of poor response to sildenafil. Methods. In a double -blinded study, 38 men aged 47 ± 1.5 years with moderate-to-severe ED and poor response to sildenafil were randomly assigned to take : pioglitazone 30 mg once daily (n=19) or placebo (n=19) for 9 weeks. Erectile function (EF) scores which were assessed by EF domain of International Index of Erectile Function (IIEF) along with responses to Global Assessment Questions (GAQs) were major outcome measures. Serum levels of total testosterone (T), dehydroepiandrosterone sulfate (DHEAS), glucose, lipid profiles and liver function tests were minor outcome measures. Results. Pioglitazone significantly improved major outcome measures compared with placebo ( p < 0.05). A decrease from baseline of total cholesterol level was more in pioglitazone group than in placebo group (p < 0.05). In 84% of the sildenafil poor responders (n=38), at least one of the associated risk factors of ED was found. There was undiagnosed hypercholesterolemia in 34% of the subjects. Serum levels of T, DHEAS, glucose and other parameters remained unchanged in both groups. Pioglitazone was well tolerated either alone or in co- administration with sildenafil. Conclusions. Pioglitazone safely increased sildenafil response to improve ED of men with prior sildenafil failure. This improvement is regardless of fasting glucose and sex hormones levels. Screening for identifying and treatment of risk factors of ED is beneficial for sildenafil poor responders

Key words:Erectile dysfunction, Pioglitazone, Sildenafil,Testosterone
Swim Training Reduces Metformin Levels in Fructose-Induced Insulin Resistant Rats
Authors: Chien Kuei-Yu , Huang Chi-Chang , Hsu Ku-Fu , Kuo Chia-Hua , Hsu Mei-Chich
Published: March 2016

Purpose: Regular exercise training and metformin medication are widely considered to increase insulin sensitivity and protect against type 2 diabetes, however, evaluating the effect of exercise training on the disposition and pharmacokinetics of metformin is unclear. Methods: We investigated the effect of a 4-wk swim training program (45 min/day, 5 days/wk) on the pharmacokinetics for the use of metformin in fructose-induced insulin resistant rats. Fructose-induced insulin resistant rats were assigned into two groups (n=6/group): swim training with metformin (SM), and non-swim training with metformin (CM). Blood samples were collected from 12 h-fasted rats at baseline and at 0.25, 0.5, 0.75, 1, 2, 3, 4, 6, 8, 10, and 12 h after an oral glucose tolerance test (OGTT) with administration of a single dose of metformin (450 mg/kg). Results: Our study revealed that both glucose and insulin levels in the SM group were significantly lower than those in the CM group at 15 min following OGTT. The maximum concentration (Cmax) and area under the serum concentration-time curve (AUC) for the SM group were significantly lower than CM group. The apparent distribution volume (Vd) and the time-averaged total body clearance (CL) for the SM group were significantly higher than those for the CM group. There were no significant differences in the time to maximum concentration (Tmax) or the time to half-life concentration (t1/2) between the two groups. Conclusions: Our data demonstrate that swim training reduces metformin serum levels.

This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.
Iron Complexation to Histone Deacetylase Inhibitors SAHA and LAQ824 in PEGylated Liposomes Can Considerably Improve Pharmacokinetics in Rats
Authors: Wang Yan , Tu Sheng , Steffen Dana , Xiong May
Published: March 2016

PURPOSE. The formulation of histone deacetylase inhibitors (HDACi) is challenging due to poor water solubility and rapid elimination of drugs in vivo. This study investigated the effects of complexing iron (Fe3+) to the HDACi suberoylanilide hydroxamic acid (SAHA) and LAQ824 (LAQ) prior to their encapsulation into PEGylated liposomes, and investigated whether this technique could improve drug solubility, in vitro release and in vivo pharmacokinetic (PK) properties. METHODS. The reaction stoichiometry, binding constants and solubility were measured for Fe complexes of SAHA and LAQ. The complexes were passively encapsulated into PEGylated liposomes and characterized by size distribution, zeta-potential, encapsulation efficiency (EE), and in vitro drug release studies. PC-3 cells were used to verify the in vitro anticancer activity of the formulations. In vivo pharmacokinetic properties of liposomal LAQ-Fe (L-LAQ-Fe) was evaluated in rats. RESULTS. SAHA and LAQ form complexes with Fe at 1:1 stoichiometric ratio, with a binding constant on the order of 104 M-1. Fe complexation improved the aqueous solubility and the liposomal encapsulation efficiency of SAHA and LAQ (29-35% EE, final drug concentration > 1 mM). Liposomal encapsulated complexes (L-HDACi-Fe) exhibited sustained in vitro release properties compared to L-HDACi but cytotoxicity on PC-3 cells was comparable to free drugs. The PK of L-LAQ-Fe revealed 15-fold improvement in the plasma t1/2 (12.11 h)and 211-fold improvement in the AUC∞ (105.7 µg·h/ml) compared to free LAQ (0.79 h, 0.5 µg·h/ml). Similarly, the plasma t1/2 of Fe was determined to be 11.83 h in a separate experiment using radioactive Fe-59. The majority of Fe-59 activity was found in liver and spleen of rats and correlates with liposomal uptake by the mononuclear phagocyte system. CONCLUSIONS. We have demonstrated that encapsulation of Fe complexes of HDACi into PEGylated liposomes can improve overall drug aqueous solubility, in vitro release and in vivo pharmacokinetic properties. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.
Selective Induction of Apoptosis by Azadarichta indica Leaf Extract by Targeting Oxidative Vulnerabilities in Human Cancer Cells
Authors: Roma Alessia , Ovadje Pamela , Steckle Matthew , Nicoletti Leah , Saleem Ammar , Pandey Siyaram
Published: March 2016

Purpose: Natural products have been a great source of medications used in conventional medicines for the treatment of various diseases; more importantly, they have played a significant role in the development of anti-cancer drugs for a number of decades. The benefits to employing whole extracts of natural health products, rather than a single ingredient, for cancer treatment remains unexplored. Our research group has previously demonstrated the potential anti-cancer benefits of several natural health products (NHPs), prompting further studies into other NHPs, such as Neem (Azadarichta indica), a tree native to India and has been used in Ayurvedic medicine for over 4000 years. The objective of this study is to determine the possible anti-cancer potential of aqueous and ethanolic Neem leaf extracts (NLEs) and to identify the specific mode(s) of action. Methods: Cells were treated with NLE and cell viability was then assessed using a water-soluble tetrazolium salt. Cell death was confirmed using the fluorescent dye propidium iodide and apoptosis was identified using the Annexin-V binding assay. Mitochondrial membrane permeabilization was visualized using JC-1 staining and the production of whole cell and mitochondrial ROS was measured with 2’,7’-dichlorodihydrofluorescein diacetate (H2DCFDA) and Amplex Red, respectively. In vivo efficacy of aqueous NLE was assessed in human tumour xenografts in CD-1 nu/nu immunocompromised mice. Results: Results indicate that both ethanolic and aqueous extracts of Neem leaf were effective in inducing apoptosis in leukemia and colon cancer cells, following destabilization of the mitochondrial membrane. Furthermore, an increase in the production of reactive oxygen species (ROS) was observed in cancer cells treated with NLEs, indicating that oxidative stress may play a role in the mechanism of cell death. Additionally, in vivo results showed that aqueous NLE (delivered orally) was well tolerated and inhibited tumour growth of human xenografts in mice. Conclusions: These findings suggest the potential of NLEs as safer and effective alternatives to conventional chemotherapy. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.
Development of a highly reproducible system to evaluate inhibition of cytochrome P450 3A4 activity by natural medicines
Authors: Sato Yu , Sasaki Takamitsu , Takahashi Shogo , Kumagai Takeshi , Nagata Kiyoshi
Published: March 2016

PURPOSE: In recent years, a number of natural medicines have been reported to have inductive or inhibitive effects on the activity of drug metabolizing enzymes, upon co-administration with prescribed medicines. However, information regarding natural medicine-drug interactions that influence drug metabolism is limited owing to the lack of efficient screening method for such interactions. Therefore, to understand whether P450 activity is affected by natural medicine in small intestines, we have established frozen recombinant P450-expressing cells infected with human CYP3A4 expressing adenovirus (Ad-CYP3A4) to evaluate the effect of natural medicines on CYP3A4 activity. METHODS: Ad-CYP3A4 cells were created by infecting HepG2 cells with Ad-CYP3A4 at 10 multiplicity of infection (MOI) and these cells were stored using cryopreservation medium (fAd-CYP3A4 cells) to obtain long-term consistent data and stable supplies of cells expressing a constant level of CYP3A4 activity. RESULTS: The CYP3A4 activity in fAd-CYP3A4 cells remained unaffected at the end of each frozen period (0, 1, 2, and 6 months). Inhibitory effect on CYP3A4 activity by typical inhibitors (ketoconazole, hyperforin) and natural medicines (Cat’s Claw, Devil’s Claw, Feverfew, Peppermint Oil, Red Clover, and Siberian Eleuthero) were evaluated. The inhibitors had nearly equal IC50 values in fAd-CYP3A4 cells, Ad-CYP3A4 cells and recombinant CYP3A4 microsomes. Cat’s Claw, Peppermint Oil and Siberian Eleuthero inhibited CYP3A4 activity more potently than 0.1 μM ketoconazole in fAd-CYP3A4 cells. CONCLUSIONS: In the present study, we have successfully developed a highly reproducible system to evaluate CYP3A4 inhibition in small intestines by natural medicines.
Preparation and Preliminary Evaluation of Novel β-Cyclodextrin/IUDR Prodrug Formulations
Authors: Wiebe Leonard I. , Yang Xiao-Hong , Singh Shradha , Diakur Jim
Published: March 2016

PURPOSE. Iododeoxyuridine (IUdR) has a very short in vivo half-life and consequently achieves low target-tissue concentrations with concomitant lower efficacy than would be predicted from in vitro studies. This work reports the preparation of IUdR:?-cyclodextrin (?-CyD) inclusion complexes designed to reduce in vivo inactivation of IUdR. METHODS. IUdR was derivatized with either 1-adamantanecarbonyl chloride or 4-(1-adamantyl-carbamoyl)butanoic acid, to prepare 5’-O-(1-adamantoyl)-5-iodo-2’-deoxyuridine 1 and 5’-O-(4-(1-adamantylcarbamoyl)butoyl)-5-iodo-2’-deoxy-uridine 4, respectively. ?-CyD complexes 5 and 6 were formed by vigorous stirring of 1:1 solutions of ?-CyD and 1 or 4, respectively, in D2O under argon. Complexation was inferred from DSC, powder x-ray diffractometry and NMR spectrometry. The dissociation of 5 in water and under cholesterol challenge, and the effect of complexation on the stability of 1 was determined by incubation in plasma. RESULTS. IUdR coupling with adamantanecarbonyl chloride proceeded smoothly to afford 1 (69 %) and the di-substituted derivative, 3’,5’-di-O-(1-adamantoyl)-5-iodo-2’-deoxyuridine 2 (8 %); 4 was obtained in 42 % yield. The formation of 1:1 complexes 5 and 6 was inferred from NMR chemical shift data. In serum, 1 was 90 % hydrolyzed to IUdR in 30 min, compared to 10 % hydrolysis of 1 to IUdR when from complex 5. CONCLUSIONS. Inclusion complexes were formed between ?-CyD and adamantamine-IUdR conjugates at 1:1 molar ratios.

The complex 5 was resistant to dissociation by cholesterol challenge, and 5 was more slowly converted to IUdR than non-complexed 1. In vivo studies are required to further exploit the ?-CyD inclusion complex approach for improved delivery of nucleoside derivatives.
Development of Lyophilized Gemini Surfactant-Based Gene Delivery Systems: Influence of Lyophilization on the Structure, Activity and Stability of the Lipoplexes
Authors: Mohammed-Saeid Waleed , Michel Deborah , El-Aneed Anas , Verrall Ronald E , Low Nicholas H , Badea Ildiko
Published: March 2016

Purpose. Cationic gemini surfactants have been studied as non-viral vectors for gene therapy. Clinical applications of cationic lipid/DNA lipoplexes are restricted by their instability in aqueous formulations. In this work, we investigated the influence of lyophilization on the essential physiochemical properties and in vitro transfection of gemini surfactant-lipoplexes. Additionally, we evaluated the feasibility of lyophilization as a technique for preparing lipoplexes with long term stability. Methods. A gemini surfactant [12-7NH-12] and plasmid DNA encoding for interferon-γ were used to prepare gemini surfactant/pDNA [P/G] lipoplexes. Helper lipid DOPE [L] was incorporated in all formulation producing a [P/G/L] system. Sucrose and trehalose were utilized as stabilizing agents. To evaluate the ability of lyophilization to improve the stability of gemini surfactant-based lipoplexes, four lyophilized formulations were stored at 25˚C for three months. The formulations were analyzed at different time-points for physiochemical properties and in vitro transfection. Results. The results showed that both sucrose and trehalose provided anticipated stabilizing effect. The transfection efficiency of the lipoplexes increased 2-3 fold compared to fresh formulations upon lyophilization. This effect can be attributed to the improvement of DNA compaction and changes in the lipoplex morphology due to the lyophilization/rehydration cycles. The physiochemical properties of the lyophilized formulations were maintained throughout the stability study. All lyophilized formulations showed a significant loss of gene transfection activity after three months of storage. Nevertheless, no significant losses of transfection efficiency were observed for three formulations after two months storage at 25 ˚C. Conclusion. Lyophilization significantly improved the physical stability of gemini surfactant-based lipoplexes compared to liquid formulations. As well, lyophilization improved the transfection efficiency of the lipoplexes. The loss of transfection activity upon storage is most probably due to the conformational changes in the supramolecular structure of the lipoplexes as a function of time and temperature rather than to DNA degradation.

This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.
Evaluation of Synthesized Cross Linked Polyvinyl Alcohol as Potential Disintegrant
Authors: Patel Ashok , Vavia Pradeep
Published: March 2016

PURPOSE: The present study deals with evaluation of crosslinked poly vinyl alcohol (PVA) as a potential disintegrant.

METHODS: Crosslinking of PVA was carried out using glutaraldehyde as a crosslinker, in presence of acidic conditions. The crosslinking reaction was optimized for a) polymer: crosslinker ratio; b) temperature requirement and c) reaction duration. Certain physical parameters of the disintegrant (including sedimentation volume, hydration capacity, specific surface area and bulk and tap density) were determined and compared to the known disintegrants. Characterization was carried out using FT-IR, DSC, XRD, SEM and Photo microscopy studies. The developed excipient was also studied for acute toxicity in rats and found to be safe for oral use.

RESULTS: Disintegration property of formed product was compared to known disintegrant (Ac-Di-Sol) and it was found to give better results. The disintegration mechanism of developed disintegrant was postulated based on results obtained from various physical evaluations including: Study of effect of disintegrant concentration, fillers, and hardness, mode of incorporation and method of granulation on disintegration activity.

CONCLUSIONS: By changing the condition parameters of well known crosslinking reaction of PVA, we obtained a crosslinked product which had excellent disintegration activity, good flow and optimal tableting properties.
Plasma Membrane Calcium ATPases as Novel Candidates for Therapeutic Agent Development
Authors: Strehler Emanuel E
Published: March 2016

Plasma membrane Ca2+ ATPases (PMCAs) are highly regulated transporters responsible for Ca2+ extrusion from all eukaryotic cells. Different PMCA isoforms are implicated in various tasks of Ca2+ regulation including bulk Ca2+ transport and localized Ca2+ signaling in specific membrane microdomains. Accumulating evidence shows that loss, mutation or inappropriate expression of different PMCAs is associated with pathologies ranging from hypertension, low bone density and male infertility to hearing loss and cerebellar ataxia. Compared to Ca2+ influx channels, PMCAs have lagged far behind as targets for drug development, mainly due to the lack of detailed understanding of their structure and specific function. This is rapidly changing thanks to integrated efforts combining biochemical, structural, cellular and physiological studies suggesting that selective modulation of PMCA isoforms may be of therapeutic value in the management of different and complex diseases. Both structurally informed rational design and high-throughput small molecule library screenings are promising strategies that are expected to lead to specific and isoform-selective modulators of PMCA function. This short review will provide an overview of the diverse roles played by PMCA isoforms in different cells and tissues and their emerging involvement in pathophysiological processes, summarize recent progress in obtaining structural information on the PMCAs, and discuss current and future strategies to develop specific PMCA inhibitors and activators for potential therapeutic applications.

This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.
MÉTODO PARA CORREÇÃO DE IMAGENS DE PHANTOM E CLASSIFICAÇÃO DE SUAS ESTRUTURAS DE INTERESSE
Authors: Barufaldi Bruno , Cavalcanti Amanda B. , Batista Leonardo V. , Góis Renata F. , Schiabel Homero , Carvalho José F. G.
Published: March 2016

O controle de câncer de mama representa um dos grandes desafios que os serviços de saúde pública enfrentam atualmente. Para realizar o controle de qualidade de sistemas mamográficos o Ministério da Saúde exige o uso de simuladores radiográficos (phantoms) de mama. Com o objetivo de reduzir a subjetividade na avaliaçáo das imagens de phantom pela inspeçáo visual humana, está sendo desenvolvido um sistema computadorizado que utiliza um método de correçáo em imagens digitalizadas, associado à classificaçáo de suas estruturas de interesse pelo critério de visibilidade. Ao comparar os resultados da classificaçáo através do algoritmo J48 da ferramenta WEKA com e sem a correçáo das imagens, essa técnica apresentou uma melhora significativa na eficácia para determinadas estruturas do phantom.
REVIEWING THE SACARWEB DESIGN BASED IN DISCOUNT USABILITY ENGINEERING
Authors: Ferracioli Fabrício , Camargo-Brune Maria Angélica de Oliveira
Published: March 2016

This work analysed the SacarWeb's project and its objectives facing a usability analysis. The objective is to verify if the software attends its objetives of learnability, errors rate, if it is usable and to integrate usability techniques in the development cycle of the SacarWeb team. For this it was tested the Discount Usability Engineering approach, that proved to be adequate to teams with low usability knowledge, since it was found many valid usability problems and coincident among different evaluation types and evaluators. Our results have shown that is possible to a team make usability inspections based in the literature, that is possible to use the usability problems as a source of tasks to development team and that is not a difficult approach to learn, so it is possible to integrate in teams without usability specialists. Additionally we conclude that usability can't be neglected in the software development cycle, since many adoption problems can occur because of this kind of problem.
PERFORMANCE OF A CAD SCHEME APPLIED TO IMAGES OBTAINED FROM MAMMOGRAPHIC FILM DIGITIZATION AND FULL-FIELD DIGITAL MAMMOGRAPHY (FFDM)
Authors: Angelo Michele Fúlvia , Schiabel Homero , Patrocinio Ana Claudia
Published: March 2016

This work has as purpose to compare the effects of a CAD scheme applied to digitized and direct digital mamograms sets. A routine designed to be applied to mammogram in DICOM standard was developed and a schema based on the Watershed Transform to masses detection was applied to 252 ROIs from 130 digitized mammograms, resulting in 92% of true positive and 10% of false positives. For clustered microcalcifications detection, another procedure was applied to 165 ROIs from 120 mammograms, resulting in 93% of true positive and 16% of false positive. By using the same procedures to 154 digital mammograms obtained from FFDM, the rates have shown a little decrease in the scheme performance: 89% of true positive and 16% of false positive for masses detection; 90% of true positive and 27% of false positive for clusters detection. Although the tests with digital mammograms have been carried with a smaller number of images and different cases compared to the digitized ones, including several dense breasts images, the results can be considered comparable, mainly for clustered microcalcifications detection with a difference of only 3% between the sensibility rates for the both images sets. Another important feature affecting these results is the contrast difference between the two images set. This implies the need of  extensive investigations not only with a larger number of cases from FFDM but also on the parameters related to its image acquisition as well as to its corresponding processing
ESCLEROSE LATERAL AMIOTRÓFICA: UMA REVISÃO DO ESTADO DA ARTE PARA SISTEMAS DE MONITORAMENTO EM HOMECARE
Authors: Morais Antonio Higor Freire de , Valentim Ricardo Alexsandro de Medeiros , Brandão Gláucio Bezerra
Published: March 2016

Esse artigo apresenta uma revisão bibliográfica do Estado da Arte dos trabalhos publicados nas bases de pesquisa do IEEE, Scielo, Elsevier e CAPES, nos últimos 2 anos, referentes aos sistemas de monitoramento homecare para Esclerose Lateral Amiotrófica (ELA). Foram encontrados 2 artigos que tratam especificamente de sistemas de monitoramento em homecare para ELA e outros 9 artigos que não tratam especificamente, mas que podem ser adaptados para tal. Diante das características dos diversos sistemas, os trabalhos apresentados oferecem diversas funções que favorecem o atendimento/cuidado de pacientes em casa, bem como a melhoria da qualidade de vida.
PROPOSTA DE UNIDADE TERAPÊUTICA DOMICILIAR DE BAIXO CUSTO BASEADA NO PROTOCOLO PEDIASUIT
Authors: Dallegrave Caroline Paola Mickosz , Neves Eduardo Borba , Gödke Francisco , Rosário Marcelo Oliveira , Costin Ana Cláudia Martins Szizypiorin , Chiarello Claudiana Renata
Published: March 2016

A Paralisia Cerebral é uma doença neuromuscular que afeta o cérebro em desenvolvimento, causando alterações sensório-motoras que podem comprometer a coordenação motora fina, a manutenção da postura e a deambulação. O uso de tecnologias assistivas específicas é de grande importância para o desenvolvimento da criança portadora de Paralisia Cerebral, uma vez que pode permitir a correção de padrões corporais errôneos e auxiliar na realização de tarefas cotidianas. Este estudo teve como objetivo descrever o desenvolvimento de uma versão de baixo custo e baixa mecanização da Unidade de Tratamento Domiciliar (UTD), baseada no protocolo Pediasuit, bem como suas vantagens e desvantagens em relação ao projeto original. Foi utilizada uma abordagem qualitativa, baseada nos pareceres de terapeutas e responsáveis após um período de testes de seis meses, durante o qual o equipamento foi utilizado por 14 crianças entre 3 e 10 anos de idade, portadoras de desordens neuromotoras. Foi constatado que o produto exige menor uso de força pelo paciente sem comprometimento do padrão da marcha. Observou-se ainda que a confecção do equipamento foi facilitada pelo uso da matéria-prima (PVC) que não exige maquinário ou acabamentos específicos, fato que também contribuiu na  redução do custo final do equipamento.