Simultaneous determination of antidementia drugs by HPLC-MS: validation data of the method and plasma level determinations in 300 patients

Aims: A rapid and simple HPLC-MS method was developed for the simultaneousdetermination of antidementia drugs, including donepezil, galantamine, rivastigmineand its major metabolite NAP 226 - 90, and memantine, for TherapeuticDrug Monitoring (TDM). In the elderly population treated with antidementiadrugs, the presence of several comorbidities, drug interactions resulting frompolypharmacy, and variations in drug metabolism and elimination, are possiblefactors leading to the observed high interindividual variability in plasma levels.Although evidence for the benefit of TDM for antidementia drugs still remains tobe demonstrated, an individually adapted dosage through TDM might contributeto minimize the risk of adverse reactions and to increase the probability of efficienttherapeutic response. Methods: A solid-phase extraction procedure with amixed-mode cation exchange sorbent was used to isolate the drugs from 0.5 mL ofplasma. The compounds were analyzed on a reverse-phase column with a gradientelution consisting of an ammonium acetate buffer at pH 9.3 and acetonitrile anddetected by mass spectrometry in the single ion monitoring mode. Isotope-labeledinternal standards were used for quantification where possible. The validatedmethod was used to measure the plasma levels of antidementia drugs in 300patients treated with these drugs. Results: The method was validated accordingto international standards of validation, including the assessment of the trueness(-8 - 11 %), the imprecision (repeatability: 1-5%, intermediate imprecision:2 - 9 %), selectivity and matrix effects variability (less than 6 %). Furthermore,short and long-term stability of the analytes in plasma was ascertained. Themethod proved to be robust in the calibrated ranges of 1 - 300 ng/mL for rivastigmineand memantine and 2 - 300 mg/mL for donepezil, galantamine and NAP226 - 90. We recently published a full description of the method (1). We found ahigh interindividual variability in plasma levels of these drugs in a study populationof 300 patients. The plasma level measurements, with some preliminaryclinical and pharmacogenetic results, will be presented. Conclusion: A simpleLC-MS method was developed for plasma level determination of antidementiadrugs which was successfully used in a clinical study with 300 patients.

Rearing method of Oryzaephilus surinamensis (L.) (Coleoptera, Silvanidae) on various wheat grain granulometry

Oryzaephilus surinamensis is one of most common insect pest of grains and a variety of stored products, and has been found in high numbers in almost all storage facilities. However, laboratory mass rearing of this insect for bioassays is not a simple task, mainly because of its feeding behavior, small size, and high mobility. Thus, the aim of this work was to develop a simple and efficient laboratory rearing method for O. surinamensis, using wheat kernels milled into different granulometry to obtain large number and standardized population at different life stages for bioassays. The adults were collected from storage grain facilities in the southern region of Brazil and 100 specimens were placed inside glass jars with wheat kernels milled at different grades and kept at 25±0.5ºC and 65±5% relative humidity. The insects were allowed to copulate and lay eggs for 10 days and then removed. The number of eggs, larvae, and pupae was counted at five-day intervals; longevity of the second generation adults was evaluated. The kernels milled at grade 20 were the best medium for offspring production: 89% of eggs by the 5th day; 30.5% larvae by the 10th day; 43% pupae by the 30th day and 63.4% adults at the 46th day. The adults survived up to 450 days. Culturing O. surinamensis under the described conditions, transferring the parental adults by the 10th day after infestation and replacing the media when population builds up will produce enough insects of each stage for various laboratory bioassays.

Calceology : an introduction to North Western European archaeological leather footwear (Neolithic period to c. AD 1600)

Calceology is the study of recovered archaeological leather footwear and is comprised of conservation, documentation and identification of leather shoe components and shoe styles. Recovered leather shoes are complex artefacts that present technical, stylistic and personal information about the culture and people that used them. The current method in calceological research for typology and chronology is by comparison with parallel examples, though its use poses problems by an absence of basic definitions and the lack of a taxonomic hierarchy. The research findings of the primary cutting patterns, used for making all leather footwear, are integrated with the named style method and the Goubitz notation, resulting in a combined methodology as a basis for typological organisation for recovered footwear and a chronology for named shoe styles. The history of calceological research is examined in chapter two and is accompanied by a review of methodological problems as seen in the literature. Through the examination of various documentation and research techniques used during the history of calceological studies, the reasons why a standard typology and methodology failed to develop are investigated. The variety and continual invention of a new research method for each publication of a recovered leather assemblage hindered the development of a single standard methodology. Chapter three covers the initial research with the database through which the primary cutting patterns were identified and the named styles were defined. The chronological span of each named style was established through iterative cross-site sedation and named style comparisons. The technical interpretation of the primary cutting patterns' consistent use is due to constraints imposed by the leather and the forms needed to cover the foot. Basic parts of the shoe patterns and the foot are defined, plus terms provided for identifying the key points for pattern making. Chapter four presents the seventeen primary cutting patterns and their sub-types, these are divided into three main groups: six integral soled patterns, four hybrid soled patterns and seven separately soled patterns. Descriptions of the letter codes, pattern layout, construction principle, closing seam placement and list of sub-types are included in the descriptions of each primary cutting pattern. The named shoe styles and their relative chronology are presented in chapter five. Nomenclature for the named styles is based on the find location of the first published example plus the primary cutting pattern code letter. The named styles are presented in chronological order from Prehistory through to the late 16th century. Short descriptions of the named styles are given and illustrated with examples of recovered archaeological leather footwear, reconstructions of archaeological shoes and iconographical sources. Chapter six presents documentation of recovered archaeological leather using the Goubitz notation, an inventory and description of style elements and fastening methods used for defining named shoe styles, technical information about sole/upper constructions and the consequences created by the use of lasts and sewing forms for style identification and fastening placement in relation to the instep point. The chapter concludes with further technical information about the implications for researchers about shoemaking, pattern making and reconstructive archaeology. The conclusion restates the original research question of why a group of primary cutting patterns appear to have been used consistently throughout the European archaeological record. The quantitative and qualitative results from the database show the use of these patterns but it is the properties of the leather that imposes the use of the primary cutting patterns. The combined methodology of primary pattern identification, named style and artefact registration provides a framework for calceological research.

Simple method for detection of MYH11 DNA rearrangements in patients with inv(16)(p13q22) and acute myeloid leukemia.

The pericentric inversion on chromosome 16 [inv(16)(p13q22)] and related t(16;16)(p13;q22) are recurrent aberrations associated with acute myeloid leukemia (AML) M4 Eo. Both abberations result in a fusion of the core binding factor beta (CBFB) and smooth muscle myosin heavy chain gene (MYH11). A selected genomic 6.9-kb BamHl probe detects MYH11 DNA rearrangements in 18 of 19 inv(16)/t(16;16) patients tested using HindIII digested DNA. The rearranged fragments were not detectable after remission in two cases tested, while they were present after relapse in one of these two cases tested.

Rate of convergence of a particle method to the solution of the Mc Kean-Vlasov's equation

This paper studies the rate of convergence of an appropriatediscretization scheme of the solution of the Mc Kean-Vlasovequation introduced by Bossy and Talay. More specifically,we consider approximations of the distribution and of thedensity of the solution of the stochastic differentialequation associated to the Mc Kean - Vlasov equation. Thescheme adopted here is a mixed one: Euler/weakly interactingparticle system. If $n$ is the number of weakly interactingparticles and $h$ is the uniform step in the timediscretization, we prove that the rate of convergence of thedistribution functions of the approximating sequence in the $L^1(\Omega\times \Bbb R)$ norm and in the sup norm is of theorder of $\frac 1{\sqrt n} + h $, while for the densities is ofthe order $ h +\frac 1 {\sqrt {nh}}$. This result is obtainedby carefully employing techniques of Malliavin Calculus.

A new alternative method for testing skin irritation using a human skin model: a pilot study.

Studies assessing skin irritation to chemicals have traditionally used laboratory animals; however, such methods are questionable regarding their relevance for humans. New in vitro methods have been validated, such as the reconstructed human epidermis (RHE) model (Episkin®, Epiderm®). The comparison (accuracy) with in vivo results such as the 4-h human patch test (HPT) is 76% at best (Epiderm®). There is a need to develop an in vitro method that better simulates the anatomo-pathological changes encountered in vivo. To develop an in vitro method to determine skin irritation using human viable skin through histopathology, and compare the results of 4 tested substances to the main in vitro methods and in vivo animal method (Draize test). Human skin removed during surgery was dermatomed and mounted on an in vitro flow-through diffusion cell system. Ten chemicals with known non-irritant (heptylbutyrate, hexylsalicylate, butylmethacrylate, isoproturon, bentazon, DEHP and methylisothiazolinone (MI)) and irritant properties (folpet, 1-bromohexane and methylchloroisothiazolinone (MCI/MI)), a negative control (sodiumchloride) and a positive control (sodiumlaurylsulphate) were applied. The skin was exposed at least for 4h. Histopathology was performed to investigate irritation signs (spongiosis, necrosis, vacuolization). We obtained 100% accuracy with the HPT model; 75% with the RHE models and 50% with the Draize test for 4 tested substances. The coefficients of variation (CV) between our three test batches were <0.1, showing good reproducibility. Furthermore, we reported objectively histopathological irritation signs (irritation scale): strong (folpet), significant (1-bromohexane), slight (MCI/MI at 750/250ppm) and none (isoproturon, bentazon, DEHP and MI). This new in vitro test method presented effective results for the tested chemicals. It should be further validated using a greater number of substances; and tested in different laboratories in order to suitably evaluate reproducibility.

Nonlinear models and small sample performance of the generalized method of moments

In this paper I explore the issue of nonlinearity (both in the datageneration process and in the functional form that establishes therelationship between the parameters and the data) regarding the poorperformance of the Generalized Method of Moments (GMM) in small samples.To this purpose I build a sequence of models starting with a simple linearmodel and enlarging it progressively until I approximate a standard (nonlinear)neoclassical growth model. I then use simulation techniques to find the smallsample distribution of the GMM estimators in each of the models.