Drug bioactivation, covalent binding to target proteins and toxicity relevance

A number of therapeutic drugs with different structures and mechanisms of action have been reported to undergo metabolic activation by Phase I or Phase II drug-metabolizing enzymes. The bioactivation gives rise to reactive metabolites/intermediates, which readily confer covalent binding to various target proteins by nucleophilic substitution and/or Schiff's base mechanism. These drugs include analgesics (e.g., acetaminophen), antibacterial agents (e.g., sulfonamides and macrolide antibiotics), anticancer drugs (e.g., irinotecan), antiepileptic drugs (e.g., carbamazepine), anti-HIV agents (e.g., ritonavir), antipsychotics (e.g., clozapine), cardiovascular drugs (e.g., procainamide and hydralazine), immunosupressants (e.g., cyclosporine A), inhalational anesthetics (e.g., halothane), nonsteroidal anti-inflammatory drugs (NSAIDSs) (e.g., diclofenac), and steroids and their receptor modulators (e.g., estrogens and tamoxifen). Some herbal and dietary constituents are also bioactivated to reactive metabolites capable of binding covalently and inactivating cytochrome P450s (CYPs). A number of important target proteins of drugs have been identified by mass spectrometric techniques and proteomic approaches. The covalent binding and formation of drug-protein adducts are generally considered to be related to drug toxicity, and selective protein covalent binding by drug metabolites may lead to selective organ toxicity. However, the mechanisms involved in the protein adduct-induced toxicity are largely undefined, although it has been suggested that drug-protein adducts may cause toxicity either through impairing physiological functions of the modified proteins or through immune-mediated mechanisms. In addition, mechanism-based inhibition of CYPs may result in toxic drug-drug interactions. The clinical consequences of drug bioactivation and covalent binding to proteins are unpredictable, depending on many factors that are associated with the administered drugs and patients. Further studies using proteomic and genomic approaches with high throughput capacity are needed to identify the protein targetsof reactive drug metabolites, and to elucidate the structure-activity relationships of drug's covalent binding to proteins and their clinical outcomes.

Clincial pharmacogenetics and potential application in personalized medicine

The current `fixed-dosage strategy' approach to medicine, means there is much inter-individual variation in drug response. Pharmacogenetics is the study of how inter-individual variations in the DNA sequence of specific genes affect drug responses. This article will highlight current  pharmacogenetic knowledge on important drug metabolizing enzymes, drug transporters and drug targets to understand interindividual variability in drug clearance and responses in clinical practice and potential use in  personalized medicine. Polymorphisms in the cytochrome P450 (CYP) family may have had the most impact on the fate of pharmaceutical drugs. CYP2D6, CYP2C19 and CYP2C9 gene polymorphisms and gene duplications account for the most frequent variations in phase I metabolism of drugs since nearly 80% of drugs in use today are metabolised by these enzymes. Approximately 5% of Europeans and 1% of Asians lack CYP2D6 activity, and these  individuals are known as poor metabolizers. CYP2C9 is another clinically significant drug metabolising enzyme that demonstrates genetic variants. Studies into CYP2C9 polymorphism have highlighted the importance of the CYP2C9*2 and CYP2C9*3 alleles. Extensive polymorphism also occurs in a majority of Phase II drug metabolizing enzymes. One of the most important polymorphisms is thiopurine S-methyl transferases (TPMT) that catalyzes the S-methylation of thiopurine drugs. With respect to drug transport  polymorphism, the most extensively studied drug transporter is  P-glycoprotein (P-gp/MDR1), but the current data on the clinical impact is limited. Polymorphisms in drug transporters may change drug's distribution, excretion and response. Recent advances in molecular research have revealed many of the genes that encode drug targets demonstrate genetic polymorphism. These variations, in many cases, have altered the targets sensitivity to the specific drug molecule and thus have a profound effect on drug efficacy and toxicity. For example, the β2-adrenoreceptor, which is encoded by the ADRB2 gene, illustrates a clinically significant genetic variation in drug targets. The variable number tandem repeat polymorphisms in serotonin transporter (SERT/SLC6A4) gene are associated with response to antidepressants. The distribution of the common variant alleles of genes that encode drug metabolizing enzymes, drug transporters and drug targets has been found to vary among different populations. The promise of pharmacogenetics lies in its potential to identify the right drug at the right dose for the right individual. Drugs with a narrow therapeutic index are thought to benefit more from pharmacogenetic studies. For example, warfarin serves as a good practical example of how pharmacogenetics can be utilized prior to commencement of therapy in order to achieve maximum efficacy and minimum toxicity. As such, pharmacogenetics has the potential to achieve optimal quality use of medicines, and to improve the efficacy and safety of both prospective and licensed drugs.

Improving English language teaching in Thailand

The teaching of English in Thailand is a matter of national concern. The national government believes that the ability of Thai people to use English for effective communication is very important for the continuing economic development of Thailand. However many students who have had primary, secondary and university exposure to English find it difficult to conduct a conversation with a native speaker of English. The reasons for this include lack of student motivation and contextual support, large classes, the dominating effects of assessment on what is taught, and the English language competency of the teachers. The research in this thesis focuses on the teaching of English as a foreign language in secondary schools in Khon Kaen. The research reported here consists of one major and three minor studies. In the major study some of the principles of action research were used to explore strategies that would improve the teaching of English in a number of secondary schools in Khon Kaen in Thailand. In the first phase of the major study I worked with two teachers to design and implement a series of classroom activities that encouraged lower secondary students to use English. In the second phase I worked with a group of teachers to design and deliver a professional development program for twenty school teachers interested in improving their English language teaching. In the third phase I used data from the first two phases to design five new activities that were used in classrooms by two teachers. Findings from the three phases indicated that working collaboratively with school teachers can be a mutually beneficial professional experience and can improve student interest and learning. In the first minor study I used interview-conversations to investigate the perceptions that subject co-ordinators and teachers have towards English language teaching. The conversations covered the merits of detailed curricula and curricula frameworks, professional development, assessment, resources, and integration of English language with other subjects. It was clear that the teachers were aware of the national government s policies for the improving English language teaching and accepted the need for change. It was equally clear that the preparation of teachers and the resources available were major limiting factors in schools to teacher effectiveness. In the second minor study I examined the teaching of Mandarin in an Australian school that suffered from some of the same resource problems as Thai schools. Although there was only one teacher available for all of the Mandarin classes in the school she was extremely effective. Her teaching was an example of best practice. It included thorough preparation, the ability to manage lessons at the pace of the learners, active classes and individual attention, detailed assessment records, and the integration of language and culture. Some or all of these could be used in Thai schools. The third minor study was an investigation of the professional development experiences of English language teachers in Thai schools. In most schools there are consultative and administrative mechanisms, acceptable to principals and teachers, in place to support professional development. Access to native speakers was seen as very important. However, the schools in Khon Kaen province have little or no access to native speakers of English. Even if they were available, the schools do not have the funds to employ them. Findings from the four studies indicate that it is quite possible to use interactive, participatory or student-centred pedagogies to teach English as a foreign language in Thai classrooms. However, one cannot expect teachers to adopt such pedagogies unless they are convinced of their value. This can be achieved most effectively through a systematic and sustained program of professional development.

Rotational and translational mobility of a highly plastic salt : Dimethylpyrrolidinium thiocyanate

Thermal analysis, impedance spectroscopy, NMR and Raman spectroscopy have been used to investigate the plastic crystal dimethylpyrrolidinium thiocyanate in order to gain further insight into the properties of organic ionic plastic crystals. This compound has a solid–solid phase transition at 82 °C, and melts at 122 °C. A step increase in conductivity of about one order of magnitude is observed at the phase transition, followed by a decrease in activation energy for conduction. A large entropy gain occurs at the II → I transition, and ¹H NMR linewidth measurements together with second moment calculations showed that the dimethylpyrrolidinium cation goes from a static state, to full isotropic tumbling. Raman measurements confirm that the cation as well as the anion exhibit increased rotational mobility when entering phase I.

Plastic crystal behaviour in tetraethylammonium dicyanamide

The plastic crystal tetraethylammonium dicyanamide ([N_2,2,2,2][dca]) has been investigated with an emphasis on structure and dynamics in the plastic phase. It was found that almost all of the volume expansion occurs at the II → I transition, with no volume expansion at the melt transition (as normally observed for crystals). The conductivity of this material shows a rapid increase at temperatures below the II → I transition, reaching values ~ 10^{− 3} S/cm in Phase I, and 0.1 S/cm in the melt. The NMR measurements show that there is a sudden onset of rotational motions of the cations at the plastic transition; below this temperature the cations appear static. The rotational motion of the cation in Phase I has been discussed in terms of isotropic tumbling.

Structure and transport properties in an N,N-substituted pyrrolidinium tetrafluoroborate plastic crystal system

The structure and transport of N-propyl-N-methylpyrrolidinium tetrafluoroborate (P₁₃BF₄) has been investigated over a wide temperature range in consequence to exhibiting properties suitable for potential solid-state superionic electrolyte applications. Prior to melting, the organic salt, P₁₃BF₄, transforms into a plastic crystal phase. Intrinsic conductivity in this solid, phase I (45–65 °C), is comparable to that in the melt (_~10^−3 S cm^−1). Ionic motion and transport properties were investigated by ¹H and ¹¹B nuclear magnetic resonance (NMR) spectroscopy. Pressure-induced plastic flow in this system may accommodate volume changes in device application and to this extent, X-ray diffraction (XRD) has been used. Scanning electron microscopy (SEM) revealed complex surface morphology and lattice imperfections associated with the strong orientational disorder of the plastic state.

Thermal and physical properties of an archetypal organic ionic plastic crystal electrolyte

The thermal and mechanical properties of the ionic plastic crystal <i>Ni>-methyl-<i>Ni>-propylpyrrolidinium hexafluorophosphate have been investigated and the effect of adding a miscible polymer on the mechanical properties is reported. The physical properties of the pure plastic crystal are discussed in detail and for the first time the change in volume with temperature for an organic ionic plastic crystal is reported. An increase in volume in conjunction with increased conductivity supports the hypothesis that ion conduction within the plastic crystal proceeds via defects. For phase I and melting, the magnitude of the volume increase does not appear to be in accord with the subtle change in conductivity. This is suggested to be due to the presence of layer defects, which allow for correlated ionic motion, which does not increase the conductivity. Addition of polymer to the plastic crystal significantly increases the mechanical strength, decreases the conductivity, but has little effect on the phase behaviour, further supporting the hypothesis of vacancy-mediated conduction.

Proton transport in choline dihydrogen phosphate/H3PO4 mixtures

Mixtures of the plastic crystal material choline dihydrogen phosphate [Choline][DHP] and phosphoric acid, from 4.5 mol% to 18 mol% H₃PO₄, were investigated and shown to have significantly higher proton conductivity compared to the pure [Choline][DHP]. This was particularly evident from the electrochemical hydrogen reduction reaction and the proton NMR diffusion measurements, in addition to ionic conductivity measured from the impedance spectroscopy. The ionic conductivity was observed to increase by more than an order of magnitude in phase I (<i>i.e.i> the highest temperature solid phase in [Choline][DHP]) reaching up to 10^&minus;2 S cm^&minus;1. The multinuclear NMR spectroscopy data suggest that, at least on the timescale of the NMR measurement, the H⁺ cations and [DHP] anions are equivalent in both phases. The pulsed field gradient NMR diffusion measurements of the 18 mol% acid sample indicate that all three ions are mobile, however the H⁺ diffusion coefficient is an order of magnitude higher than for the [Choline] cation or the [DHP] anion, and therefore conduction in these materials is dominated by proton conductivity. The thermal stability, as measured by TGA, is unaffected with increasing acid additions and remains high; <i>i.e.i> no significant mass loss below 200 °C.

Disposition pathways and pharmacokinetics of herbal medicines in humans

Pharmacokinetic studies have become an integral part of modern drug development, but these studies are not regulatory needs for herbal remedies. This paper updates our current knowledge on the disposition pathways and pharmacokinetic properties of commonly used herbal medicines in humans. To retrieve relevant data, the authors have searched through computer-based literatures by full text search in Medline (via Pubmed), ScienceDirect, Current Contents Connect (ISI), Cochrance Library, CINAHL (EBSCO), CrossRef Search and Embase (all from inception to May 2010). Many herbal compounds undergo Phase I and/or Phase II metabolism in vivo, with cytochrome P450s (CYPs) and uridine diphosphate glucuronosyltransferases (UGTs) playing a major role. Some herbal ingredients are substrates of P-glycoprotein (P-gp) which is highly expressed in the intestine, liver, brain and kidney. As such, the activities of these drug metabolizing enzymes and drug transporters are determining factors for the in vivo bioavailability, disposition and distribution of herbal remedies. There are increasing pharmacokinetic studies of herbal remedies, but these studies are mainly focused on a small number of herbal remedies including St John's wort, milk thistle, sculcap, curcumin, echinacea, ginseng, ginkgo, and ginger. The pharmacokinetic data of a small number of purified herbal ingredients, including anthocyanins, berberine, catechins, curcumin, lutein and quercetin, are available. For the majority of herbal remedies used in folk medicines, data on their disposition and biological fate in humans are lacking or in paucity. For a herbal medicine, the pharmacological effect is achieved when the bioactive agents or the metabolites reach and sustain proper levels at their sites of action. Both the dose levels and fates of active components in the body govern their target-site concentrations after administration of an herbal remedy. In this regard, a safe and optimal use of herbal medicines requires a full understanding of their pharmacokinetic profiles. To optimize the use of herbal remedies, further clinical studies to explore their biological fate including the disposition pathways and kinetics in the human body are certainly needed.

Lithium doped N,N-dimethyl pyrrolidinium tetrafluoroborate organic ionic plastic crystal electrolytes for solid state lithium batteries

The organic ionic plastic crystal material N,N-dimethyl pyrrolidinium tetrafluoroborate ([C1mpyr][BF4]) has been mixed with LiBF4 from 0 to 8 wt% and shown to exhibit enhanced ionic conductivity, especially in the higher temperature plastic crystal phases (phases II and I). The materials retain their solid state well above 100 °C with the melt not being observed up to 300 °C. Interestingly the conductivity enhancement is highest with the lowest level of LiBF4 addition in phase II, but then the order of enhancement is reversed in phase I. In all cases, a conductivity drop is observed at the II → I phase transition (105 °C) which is associated with increased order in the pure matrix, as previously reported, although the conductivity drop is least for the highest LiBF4 amount (8 wt%). The 8 wt% sample displays different conductivity behaviours compared to the lower LiBF4 concentrations, with a sharp increase above 50 °C, which is apparently not related to the formation of an amorphous phase, based on XRD data up to 120 °C. Symmetric cells, Li/OIPC/Li, were prepared and cycled at 50 °C and showed evidence of significant preconditioning with continued cycling, leading to a lower over-potential and a concomitant decrease in the cell resistivity as measured by EIS. An SEM investigation of the Li/OIPC interfaces before and after cycling suggested significant grain refinement was responsible for the decrease in cell resistance upon cycling, possibly as a result of an increased grain boundary phase.

Neighbourhood environments and children's social capital : investigating the pattern of relationships

The main purpose of this work was to investigate the pattern of relationships among three constructs: neighbourhood socio-physical environment, children's social interactions and their social capital. This work was designed as a two-phase mixed-methods research. Phase I included several qualitative studies to develop a scale of neighbourhood socio-physical environment, a scale of children's social interactions and a scale of children's social capital. Phase II was a cross-national survey that used these three scales to collect information from high school students in Beijing and Sydney. The main finding of this work was that there were strong and significant correlations among the three constructs. Children's assessment of their neighbourhood socio-physical environment was positively correlated with their social interactions and social capital, which indicated that children who lived in better neighbourhoods had more social interactions and larger volumes of social capital. Strong positive relationship was also found between children's social interactions and social capital, which implied that better-connected children interacted with their friends more.

Hormonal responses to extreme fasting in subantarctic fur seal (Arctocephalus tropicalis) pups

Surviving prolonged fasting implies closely regulated alterations in fuel provisioning to meet metabolic requirements, while preserving homeostasis. Little is known, however, of the endocrine regulations governing such metabolic adaptations in naturally fasting free-ranging animals. The hormonal responses to natural prolonged fasting and how they correlate to the metabolic adaptations observed, were investigated in subantarctic fur seal (Arctocephalus tropicalis) pups, which, because of the intermittent pattern of maternal attendance, repeatedly endure exceptionally long fasting episodes throughout their development (1–3 mo). Phase I fasting was characterized by a dramatic decrease in plasma insulin, glucagon, leptin, and total L-thyroxine (T4) associated with reductions in mass-specific resting metabolic rate (RMR), plasma triglycerides, glycerol, and urea-to-creatine ratio, while nonesterified fatty acids (NEFA) and β-OHB increased. In contrast, the metabolic steady-state of phase II fasting reached within 6 days was associated with minimal concentrations of insulin, glucagon, and leptin; unchanged cortisol and triiodothyronine (T3); and moderately increased T4. The early fall in insulin and leptin may mediate the shift to the strategy of energy conservation, protein sparing, and primary reliance on body lipids observed in response to the cessation of feeding. In contrast to the typical mammalian starvation response, nonelevated cortisol and minimal glucagon levels may contribute to body protein preservation and downregulation of catabolic pathways, in general. Furthermore, thyroid hormones may be involved in a process of energy conservation, independent of pups' nutritional state. These original hormonal settings might reflect an adaptation to the otariid repeated fasting pattern and emphasize the crucial importance of a tight physiological control over metabolism to survive extreme energetic constraints.

Capecitabine and oral cyclophosphamide : a novel oral treatment combination for advanced cancer

Background: This dose escalation study assessed feasibility of a totally oral chemotherapy regimen using cyclophosphamide and capecitabine. The rationale for this combination was based on the observation that preclinical models of cyclophosphamide up-regulated tumor thymidine phosphorylase and increased the activation of capecitabine. Methods: Eligible patients with advanced cancer were treated with oral cyclophosphamide and capecitabine on a 28-day cycle. If no dose limiting toxicities (DLT) were encountered during the first two treatment cycles, the next patient group was assigned to the next highest dose level until the maximum tolerable dose (MTD) was determined. Results: Twenty-seven patients entered treatment. The majority of non-DLT were grades 1 and 2. DLT experienced in the first 8-week observation period were grade 3 diarrhea (one patient, level III) and grade 3 emesis (two patients, level V). MTD was observed at level 5, 1331 mg/m2/day capecitabine days 1–28 with 125 mg/m2/day cyclophosphamide days 1–14 of the 28-day cycle. The recommended phase II dose is therefore 1331 mg/m2/day capecitabine with 100 mg/m2/day cyclophosphamide. The best response evaluation showed four partial responses (breast, colon, ovary and pancreas). Conclusion: Cyclophosphamide and capecitabine can be combined at their full oral single agent dose with promising tolerability and activity.