Circum-Mediterranean phylogeography of a bat coupled with past environmental niche modeling: A new paradigm for the recolonization of Europe?

The isolation of populations in the Iberian, Italian and Balkan peninsulas during the ice ages define four main paradigms that explain much of the known distribution of intraspecific genetic diversity in Europe. In this study we investigated the phylogeography of a wide-spread bat species, the bent-winged bat, Miniopterus schreibersii around the Mediterranean basin and in the Caucasus. Environmental Niche Modeling (ENM) analysis was applied to predict both the current distribution of the species and its distribution during the last glacial maximum (LGM). The combination of genetics and ENM results suggest that the populations of M. schreibersii in Europe, the Caucasus and Anatolia went extinct during the LGM, and the refugium for the species was a relatively small area to the east of the Levantine Sea, corresponding to the Mediterranean coasts of present-day Syria, Lebanon, Israel, and northeastern and northwestern Egypt. Subsequently the species first repopulated Anatolia, diversified there, and afterwards expanded into the Caucasus, continental Europe and North Africa after the end of the LGM. The fossil record in Iberia and the ENM results indicate continuous presence of Miniopterus in this peninsula that most probably was related to the Maghrebian lineage during the LGM, which did not persist afterwards. Using our results combined with similar findings in previous studies, we propose a new paradigm explaining the general distribution of genetic diversity in Europe involving the recolonization of the continent, with the main contribution from refugial populations in Anatolia and the Middle East. The study shows how genetics and ENM approaches can complement each other in providing a more detailed picture of intraspecific evolution.

Evaluation and enhancement of physical stability of semi-solid dispersions containing piroxicam into hard gelatin capsules.

The aim of the study was to investigate the physical stability of the semi-solid dispersions into the hard gelatine capsules prepared with Gelucire 44/14, Labrasol and different additives such as microcrystalline cellulose (MCC), mannitol and lactose (alpha-monohydrate) used for enhancing the stability of the formulations. The master dispersion prepared with only Gelucire 44/14 (20% w/w) and Labrasol (80% w/w) was stored in a refrigerator (5 +/- 3 degrees C), while the modified dispersions with the additives (2% w/w) were kept in a climatic chamber (25 +/- 2 degrees C / 60 +/- 5% RH) for 12 months. Dissolution tests of the semi-solid dispersions were performed in media with different pH's immediatly after preparation and after 3, 6 and 12 months of storage. FTIR and DSC studies were also carried out at the same time points. The ideal storage condition for the master dispersion was found to be at 5 degrees C. The addition of MCC, mannitol and lactose (alpha-monohydrate) to the original dispersion afforded a solidification effect on the formulation at room temperature and showed the same dissolution behavior (not less than 85% of piroxicam within 30 min in pH 1.2, 4.5 and 6.8; and water) with the master. The dispersion including lactose was stable at 25 degrees C for 12 months. However, the ideal period of storage for the modified dispersions including MCC and mannitol was 6 months at 25 degrees C. FTIR and DSC results both confirmed the amorphous state of piroxicam in all semi-solid dispersions under storage conditions for 12 months.

Nanoscale Delivery Systems of Lutein: An Updated Review from a Pharmaceutical Perspective.

Carotenoids are natural lipid-soluble pigments that produce yellow to red colors in plants as well as providing bright coloration in vegetables and fruits. Lutein belongs to the xanthophyll subgroup of the carotenoid family, which plays an essential role in photosynthesis and photoprotection in nature. In the human body, lutein, together with its isomer zeaxanthin and its metabolite meso-zeaxanthin, accumulates in the macula of the eye retina, which is responsible for central, high-resolution, and color vision. As a bioactive phytochemical, lutein has essential physiological functions, providing photoprotection against damaging blue light, along with the neutralization of oxidants and the preservation of the structural and functional integrity of cellular membranes. As a potent antioxidant and anti-inflammatory agent, lutein unfortunately has a low bioavailability because of its lipophilicity and a low stability as a result of its conjugated double bonds. In order to enhance lutein stability and bioavailability and achieve its controlled delivery to a target, nanoscale delivery systems, which have great potential for the delivery of bioactive compounds, are starting to be employed. The current review highlights the advantages and innovations associated with incorporating lutein within promising nanoscale delivery systems, such as liposomes, nanoemulsions, polymer nanoparticles, and polymer-lipid hybrid nanoparticles, as well as their unique physiochemical properties.

Poly (epsilon-caprolactone) microparticles containing Levobunolol HCl prepared by a multiple emulsion (W/O/W) solvent evaporation technique: effects of some formulation parameters on microparticle characteristics.

The aim of this study was to prepare poly (epsilon-caprolactone) (PCL) microparticles of Levobunolol HC1 (L-HC1) for use as an anti-glaucomatous drug to the eye. The double emulsion (W/O/W) solvent evaporation technique was used for encapsulating L-HC1 as a hydrophilic drug. The study examined the impact of different factors including the pH and volume of the external aqueous phase, the concentration of polyvinylalcohol (PVA) and Pluronic F68 (PF68) used as stabilizers and drug/polymer ratios on the characteristics of the microparticles. Scanning electron microscopy (SEM) and differential scanning calorimetry (DSC) were used to identify the physical state of the drug and polymer. The zeta potential of the particles was also identified. Entrapment efficiency was found to be highest with a 0.5% PVA concentration and 100 mL volume of external aqueous phase at pH 12. The high efficiency was due to a reduction in the degree of drug ionization. The microparticles were spherical and appropriately sized for ophthalmic application. Drug release from the microparticles appears to consist of two components, with an initial rapid release followed by a slower stage. Drug release was slower when the microparticle was incorporated into the thermally reversible gel (Pluronic F127) in comparison to drug release from the free drug incorporated into the gel and drug release from the free microparticle.

Water-immiscible room temperature ionic liquids (RTILs) as drug reservoirs for controlled release.

Water-immiscible room temperature ionic liquids (RTILs) have a largely unexplored potential as pharmaceutical solvents and reservoirs. This paper explores some relevant properties of the hexafluorophosphate (PF6(-)) salts of butyl, hexyl and octyl-3-methylimidazolium cations (BMIM, HMIM, OMIM, respectively). The dodecyl analogue is solid at room temperature, but its melting point can be lowered by addition of the lower homologues. Although water-immiscible, the liquids absorb water to an extent depending on their structure, the higher alkyl analogues having a lower affinity for absorbed water. The RTIL/water partition coefficients of sucrose, penicillin V potassium, dexametasone, progesterone and dehydro-epiandrosterone have been compared with octanol-water coefficients. The viscosities of the salts were measured in anhydrous, water-saturated and intermediate states. The PF6(-) ionic liquids display a low and decreasing aqueous solubility as the alkyl chain length is increased: 0.035 moll(-1) (BMIM), 0.032 moll(-1) (HMIM) and 0.09 moll(-1) (OMIM). The release of sucrose and dexametasone from RTIL reservoirs into water can be prolonged over 48 h. Saturated solutions of these RTILs show little toxicity towards Caco-2 cells, although the OMIM derivative, which is more surface-active, has a small effect on cell viability.

Template Synthesis of Tubular Nanostructures for Loading Biologically Active Molecules.

The template synthesis is a low cost, simple and versatile nanofabrication method to produce cylindrical/tubular nanostructures with controllable dimensions such as length, diameter and aspect ratio. This method utilizes nanoporous membranes such as anodized aluminum oxide (AAO) or polycarbonate (PC) as templates which have nanosized specific, cylindrical and uniform inner pores to be coated with the desired material. Template synthesized nanotubular structures have been produced from variety of materials including ceramics, polymers and proteins for loading biologically active molecules. Available procedures of material deposition into the template nanopores consist of several techniques like wetting (melt or solution wetting), layer-by-layer (LbL) assembly and sol-gel chemistry. Template synthesis enables not only control of the geometry of the resulting nanostructures but also provides nanovehicles having separated inner and outer surfaces which can be variously functionalized. Tubular nanostructures fabricated by this method have numerous potential applications including delivery of biologically active molecules such as drugs, gene, enzymes and proteins. In this review we aimed to present up-to-date works on the template based synthesis which has greatly facilitated the fabrication of polymer and protein tubular nanostructures, principally. The strategies regarding the synthesis and designing of these promising tubular nanostructures together with recent approaches relevant of drug delivery was also presented.

Acute myocardial infarction following an arthropod bite: is hereditary thrombophilia a contributing factor?

Acute myocardial infarction (AMI) due to arthropod envenomation has rarely been reported in the literature. In the present report, we describe two cases who developed AMI following an arthropod bite. Coronary angiograms revealed normal coronary arteries in both patients. Both events were probably secondary to coronary artery thrombosis and/or coronary artery vasospasm. Both patients were subsequently found to be heterozygous for prothrombin mutation (G20210A). As a result, we recommend ruling out the possibility of hereditary thrombophilias in young patients with AMI developing after an arthropod bite.

A selective and sensitive stability-indicating HPLC method for the validated assay of etoposide from commercial dosage form and polymeric tubular nanocarriers.

Etoposide is a topoisomerase II enzyme inhibitor type chemotherapeutic agent which is widely used in the therapy of various cancers. Its short half-life and toxicity to normal tissues are the major drawbacks in its clinical applications. Polymeric nanoparticulate drug delivery systems are rational carriers to deliver etoposide with higher efficiency and fewer side effects. In addition tubular shaped drug carriers are found to show a great potential for drug delivery on the basis of promising results regarding particle shape and cellular uptake. In this study, etoposide loaded polymeric tubular nanocarriers have been developed by template wetting method using porous anodic aluminum oxide membranes as templates. The developed poly(methyl methacrylate) nanocarriers were evaluated for structural analysis, in vitro drug release studies and drug release kinetics. Accurate and reliable determination of the drug release from newly developed nanocarriers, is of great importance. For this reason a selective and sensitive reversed phase liquid chromatography method was developed and fully validated from the point of system suitability, specificity, linearity and range, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy and robustness for the reliable determination of etoposide. Stability indicating capability was shown with forced degradation studies and the chromatographic conditions were optimized on ACE 5C18 (150 mm × 4.6mm I.D., 5 µm) analytical column. Related to the calibration results ETP was found linear in the range between 0.2 from 100 µg mL(-1) with the LOD as 0.015 µg.mL(-1). The resultant conditions were applied for the selective and sensitive determination of etoposide from its commercial dosage form with the high accuracy values (99.82-100.65%). The method was successfully detected assay of etoposide release from newly developed polymeric tubular nanocarriers, which was found as 72.2% at the end of 24h.

Ofloxacin Loaded Electrospun Fibers for Ocular Drug Delivery: Effect of Formulation Variables on Fiber Morphology and Drug Release.

Ofloxacin (OFL) loaded poly(ε-caprolactone) (PCL) and PCL: poly(butylene succinate) PBS fibers as a drug delivery system in the treatment of ocular infections were prepared by electrospinning. In particular, the effect of some formulation variables including polymer:drug ratio (9:1, 8:2 and 7:3 w/w), solvent systems like dichloromethane (DCM), N,N-dimethylformamide (DMF), N,Ndimethylacetamide (DMAc) and dimethylsulfoxide (DMSO), polymer blends of PCL:PBS at 80:20, 60:40 and 40:60 ratios on fiber morphology, fiber size were investigated. The morphology and diameter of the electrospun fibers were investigated by scanning electron microscopy (SEM) images also the thermal properties were evaluated by differential scanning calorimetry (DSC). The drug release behaviour from fibers and in vitro antibacterial activity were also studied. It was noticed that the average fiber diameter decreased with decreasing polymer amount in initial composition meanwhile the release of drug increased with increasing amount of drug in formulations. Solvent system of DCM:DMF at 80:20 ratio improved fiber morphology and resulted in a reduction in fiber diameter. It was found that smooth surface, flexible fibers with uniform morphology were obtained with 80:20 ratio of PCL:PBS compositions. All fibers showed a burst release of OFL. The initial amount of the released OFL was found to vary as a function of PCL:OFL ratio and polymer composition in the fiber. The microbiological activity of optimized formulation was evaluated using P. aeruginosa, S. epidermidis, S. Aureus and E. coli strains and the results of this study clearly demonstrated that freely released OFL from fibers inhibited the growth of the tested bacteria. The process of electrospinning had no adverse effect on the activity of incorporated drug in fibers.

Improved solubility and dissolution rate of piroxicam using gelucire 44/14 and labrasol.

Piroxicam is a nonsteroidal anti-inflammatory drug that is characterized by low solubility-high permeability. The present study was designed to improve the dissolution rate of piroxicam at the physiological pH's through its increased solubility by preparing semi-solid dispersions of drug using Gelucires and Labrasol. These excipients are essentially characterized by their melting points and HLB (hydrophilic-lipophilic balance) values. The dissolution tests of the preparations were performed in the media with different pH's. Differential scanning calorimetry (DSC), were used to examine the interaction between piroxicam and excipients. Gelucire 44/14 and Labrasol at the concentration of 15% w/v in water provided 20- and 50-fold increase in the solubility of piroxicam, respectively. The semi-solid dispersion containing 1/20 of drug/excipient mixture (20% Gelucire 44/14 and 80% Labrasol in w/w) produced the dissolution not less than 85% of piroxicam within 30 min in each dissolution media (simulated gastric fluid (SGF), pH 1.2; phosphate buffers, pH 4.5 and 6.8; and water). DSC analysis of this semi-solid dispersion indicated that there was no chemical reaction between the drug and excipients, and that a solid-state solution of piroxicam with excipient formed.

Enhanced bioavailability of piroxicam using Gelucire 44/14 and labrasol: in vitro and in vivo evaluation.

Piroxicam is a non-steroidal anti-inflammatory drug that is characterized by low solubility and high permeability. The purpose of the study was to investigate the in vitro and in vivo performance of the semi-solid dispersion prepared with Gelucire 44/14 and Labrasol into hard gelatin capsules (GL) for enhancing the dissolution rate of the drug. The results were evaluated by comparing with pure piroxicam filled into hard gelatin capsules (PP) and a commercially available tablet dosage form containing a piroxicam:beta-cyclodextrin complex (CD). The in vitro dissolution testing of the dosage forms was performed in different media (simulated gastric fluid, pH 1.2; phosphate buffers, pH 4.5 and 6.8; and water). Amongst the dosage forms, GL provided at least 85% piroxicam dissolution within 30 min in each of the media, behaving like a fast-dissolving immediate release drug product. Oral bioavailability of 20 mg piroxicam in GL, CD, and PP was compared after administration of a single dose to eight healthy volunteers. Three treatments were administered in crossover fashion, separated by a washout period of 2 weeks. Piroxicam was monitored in plasma by high-performance liquid chromatography. The apparent rate of absorption of piroxicam from GL (Cmax=2.64 micrograms/ml, tmax=82.5 min) was significantly higher than that of the PP (Cmax=0.999 micrograms/ml, tmax=144 min) (P<0.05) and similar to that of CD (Cmax=2.44 micrograms/ml, tmax=120 min) (P>0.05). The relative bioavailability values as the ratios of mean total AUC for GL relative to PP and CD, were 221 and 98.6%. Piroxicam is characterized by a slow and gradual absorption via the oral route and this causes a delayed onset of therapeutic effect. Thus, plain piroxicam preparations are not indicated for analgesia. The results of the in vivo study revealed that the GL dosage form would be advantageous with regards to rapid onset of action, especially in various painful conditions where an acute analgesic effect is desired.

A new record for Turkish lice fauna: Dennyus hirundinis (Linnaeus, 1761) (Mallophaga: Menoponidae).

In this study, a bird louse Dennyus hirundinis was recorded for the first time from the swift, Apus apus living in Turkey. Since so little information is available on this rare species, the new record is briefly presented here.

Comparative evaluation of granules made with different binders by a fluidized bed method.

Granules were prepared using three different binders, pregelatinized starch (PGS), gelatin (GEL), and polyvinylpyrrolidone (K30) by a fluidized bed method. As a quantitative measurement of mechanical strength or abrasion resistance, granules were subjected to a friability test for certain periods of time, and friability indexes (FI) as a function of time were calculated. The data obtained were analyzed by applying standard mathematical models. According to the derived parameters of the logistic and Weibull models, which fit best to the data, mechanical strength of granules made with K30 was observed to be lower than that of the granules of PGS and GEL which have similar values of model parameters. Flow properties, consolidation, and compressibility behaviors of unfriabled (UFR) and friabled (FR) granules, which were selected based upon their Weibull time parameter, were investigated as comparative. The flow rate of granules decreased due to diminishing particle size depending on binder type and friability, but the values of angle of repose were within the acceptable limits. Regarding consolidation behavior, the change of relative density vs. the number of taps, i.e., packing rate for FR granules of GEL was slower than that of its UFR form, whereas FR granules of PGS and K30 showed faster change in relative density compared to their UFR forms. According to the parameters obtained from the Heckel equation, PGS and K30 were found to produce softer, more plastic and readily deformable granules than GEL, and the compressibility of their FR forms was not influenced negatively.