Timing the Emergence of Resistance to Anti-HIV Drugs with Large Genetic Barriers

New antiretroviral drugs that offer large genetic barriers to resistance, such as the recently approved inhibitors of HIV-1 protease, tipranavir and darunavir, present promising weapons to avert the failure of current therapies for HIV infection. Optimal treatment strategies with the new drugs, however, are yet to be established. A key limitation is the poor understanding of the process by which HIV surmounts large genetic barriers to resistance. Extant models of HIV dynamics are predicated on the predominance of deterministic forces underlying the emergence of resistant genomes. In contrast, stochastic forces may dominate, especially when the genetic barrier is large, and delay the emergence of resistant genomes. We develop a mathematical model of HIV dynamics under the influence of an antiretroviral drug to predict the waiting time for the emergence of genomes that carry the requisite mutations to overcome the genetic barrier of the drug. We apply our model to describe the development of resistance to tipranavir in in vitro serial passage experiments. Model predictions of the times of emergence of different mutant genomes with increasing resistance to tipranavir are in quantitative agreement with experiments, indicating that our model captures the dynamics of the development of resistance to antiretroviral drugs accurately. Further, model predictions provide insights into the influence of underlying evolutionary processes such as recombination on the development of resistance, and suggest guidelines for drug design: drugs that offer large genetic barriers to resistance with resistance sites tightly localized on the viral genome and exhibiting positive epistatic interactions maximally inhibit the emergence of resistant genomes.

Y3-xBa3+xCu6O14+δ system: A new family of superconductors

Compounds of the Y3-x Ba3+x Cu6O14+δ system, which YBa2Cu3O7-δ (x = 1) is member, have been prepared. A relatively low temperature nitrate decomposition method gives almost single phase compounds with tetragonal structure. The phases are metastable and show superconducting transitions (zero-resistance) around 50K.

Understanding and management of resistance to Group M, L and I Herbicides

This project will develop better understanding of resistance to glyphosate, paraquat and Group I herbicides to better inform weed management. The project will develop a range of tools for farm advisors to improve their confidence in decision making with respect to reducing the risk of glyphosate, Group I and paraquat resistance. These will include risk assessments, case studies and scenario exploring tools. The project will discuss with commercial providers the potential for new herbicide registrations. The project will establish farm advisor learning groups to work on the application of the research in local areas where resistance is already a major problem and to improve adoption of research outcomes from this and other projects.

Germination Biology and Occurrence of Polyembryony in Two Forms of Cats Claw Creeper Vine, Dolichandra unguis-cati (Bignoniaceae): Implications for Its Invasiveness and Management

Cat’s claw creeper vine, Dolichandra unguis-cati (L.) Lohmann (syn. Macfadyena unguis-cati (L.) Gentry), is a major environmental weed in Australia. Two forms of the weed with distinctive leaf morphology and reproductive traits, including varying fruit size, occur in Queensland, Australia. The long pod form occurs in a few localities in Queensland, while the short pod form is widely distributed in Queensland and northern part of New South Wales. This investigation aimed to evaluate germination behavior and occurrence of polyembryony (production of multiple seedlings from a single seed) in the two forms of the weed. Seeds were germinated in growth chambers set to 10/20°C, 15/25°C, 20/30°C, 30/45°C and 25°C, representing ambient temperature conditions of the region. Germination and polyembryony were monitored over a period of 12 weeks. For all the treatments in this study, seeds from short pod plants exhibited significantly higher germination rates and higher occurrence of polyembryony than those from long pod plants. Seeds from long pod plants did not germinate at the lowest temperature of 10/20°C; in contrast, those of the short pod form germinated under this condition, albeit at a lower rate (reaching a maximum 45% germination at week 12). Results from this study could explain why the short pod form of D. unguis-cati is the more widely distributed plants in Australia, while the long pod is confined to a few localities. The results have implication in predicting future range of both forms of the invasive D. unguis-cati, as well as inform management decisions for control of the weed.

The high- and low-activity forms of human plasma phospholipid transfer protein (PLTP)

Plasma phospholipid transfer protein (PLTP) plays a crucial role in high-density lipoprotein (HDL) metabolism and reverse cholesterol transport (RCT). It mediates the generation of pre-beta-HDL particles, enhances the cholesterol efflux from peripheral cells to pre-beta-HDL, and metabolically maintains the plasma HDL levels by facilitating the transfer of post-lipolytic surface remnants of triglyceride-rich lipoproteins to HDL. In addition to the antiatherogenic properties, recent findings indicate that PLTP has also proatherogenic characteristics, and that these opposite characteristics of PLTP are dependent on the site of PLTP expression and action. In human plasma, PLTP exists in a high-activity (HA-PLTP) and a low-activity form (LA-PLTP), which are associated with macromolecular complexes of different size and composition. The aims of this thesis were to isolate the two PLTP forms from human plasma, to characterize the molecular complexes in which the HA- and LA-PLTP reside, and to study the interactions of the PLTP forms with apolipoproteins (apo) and the ability of apolipoproteins to regulate PLTP activity. In addition, we aimed to study the distribution of the two PLTP forms in a Finnish population sample as well as to find possible regulatory factors for PLTP by investigating the influence of lipid and glucose metabolism on the balance between the HA- and LA-PLTP. For these purposes, an enzyme-linked immunosorbent assay (ELISA) capable of determining the serum total PLTP concentration and quantitating the two PLTP forms separately was developed. In this thesis, it was demonstrated that the HA-PLTP isolated from human plasma copurified with apoE, whereas the LA-PLTP formed a complex with apoA-I. The separation of these two PLTP forms was carried out by a dextran sulfate (DxSO4)-CaCl2 precipitation of plasma samples before the mass determination. A similar immunoreactivity of the two PLTP forms in the ELISA could be reached after a partial sample denaturation by SDS. Among normolipidemic Finnish individuals, the mean PLTP mass was 6.6 +/- 1.5 mg/l and the mean PLTP activity 6.6 +/- 1.7 umol/ml/h. Of the serum PLTP concentration, almost 50% represented HA-PLTP. The results indicate that plasma HDL levels could regulate PLTP concentration, while PLTP activity could be regulated by plasma triglyceride-rich very low-density lipoprotein (VLDL) concentration. Furthermore, new evidence is presented that PLTP could also play a role in glucose metabolism. Finally, both PLTP forms were found to interact with apoA-I, apoA-IV, and apoE. In addition, both apoE and apoA-IV, but not apoA-I, were capable of activating the LA-PLTP. These findings suggest that the distribution of the HA- and LA-PLTP in human plasma is subject to dynamic regulation by apolipoproteins.

Development of Novel Assays for Measuring Different Molecular Forms of Prostate Specific Antigen

Prostate cancer (PCa) is the most commonly diagnosed non-skin cancer and second leading cause of cancer-related death of men in developed countries. Measurement of prostate specific antigen (PSA) is a very sensitive method for diagnosing and monitoring of prostate cancer (PCa), but the specificity needs improvement. Measurements of different molecular forms of PSA have been shown to improve differentiation between PCa and benign prostatic diseases. However, accurate measurement of some isoforms has not been achieved in previous assays. The aim of the present study was to develop new assays that reliably measure enzymatically active PSA, PSA-α1-chymotryposin (PSA-ACT) and PSA-α1-protease inhibitor (PSA-API), and to evaluate their diagnostic value. Double-label immunofluorometric assays using a novel monoclonal antibody (MAb) and another antibody to either free PSA (fPSA) or total PSA (tPSA) were developed and used to measure PSA-ACT and fPSA or tPSA at the same time. These assays provide enough sensitivity for measurement of PSA-ACT in sera with low PSA levels. The results obtained confirmed that proportion of PSA-ACT to tPSA (%PSA-ACT) was as useful as proportion of fPSA to tPSA (%fPSA) for discrimination between PCa and benign prostatic hyperplasia (BPH). We developed an immunoassay for detection of PSA-API based on proximity ligation, which improved assay sensitivity 10-fold compared with conventional assays. Our results confirmed previous findings that the PSA-API level is somewhat lower in men with than without PCa, and the combination of %fPSA and proportion of PSA-API to tPSA (%PSA-API) provides diagnostic improvement compared with either method alone. Assays based on this principle should be applicable to other immunoassays in which the nonspecific background is a problem. An immunopeptidometric sandwich assay (IPMA) was developed to measure the enzymatically active PSA. This assay showed high specificity, but sensitivity was not good enough for measurement of PSA concentrations in the gray zone, 2-10 µg/L, in which tPSA does not efficiently differentiate between PCa and BPH. We further developed a solid-phase proximity ligation immunoassay, which provided a 10-fold improvement in sensitivity. This proof of concept study shows that peptides reacting with proteins are potentially useful for sensitive and specific measurement of protein variants for which specific MAbs cannot be obtained.

ABOUT FORMS OF ELECTRONIC AND IONIC DEVICES WITH THERMIONIC CATHODES.

The miniaturization of electronic and ionic devices with thermionic cathodes and thc improvement of their vacuum properties are questions of very great interest to the electronic engineer. However there have bcen no proposals so far to analyse the problem of miniaturization of such devices In a fundamental way. The present work suggests a choice of the geometrical shape of the cathode, the anode and the envelope of the device, that may help towards such a fundamcnlal approach.It is shown that a design, in which the cathode and the envelope of the tube are made of thm prismatic shape and the anode coincides with the cnvclope, offers a slriknrg advantage over the conventional cylindrical design, in respect of over-all size. The use of the prismatic shape will lead to considerable economy in msterials and may facilitate simpler prodoct~ont echn~ques. I n respect of the miin criteria of vacuum, namely the grade of vacuum, the internal volume occupied by residual gases, the evolution of gases in the internal space and the diffusion of gases from outside into the devicc, it is shown that the prismatic form is at least as good as, if not somewhat superior lo, the cylindrical form.In the actual construction of thin prismatic tubes, manv practical problems will arise, the most important being the mechanical strength and stablity of the structure. But the changeover from the conventional cylindrical to the new prirmaiic form, with its basic advantages, is a development that merits close attention.

Descriptions of new genera and species of microlepidoptera from Panama (with one plate)

The following decriptions [sic] of new forms of Microlepidoptera are published in advance of proposed papers, dealing with the lepidopterous fauna of Panama as a whole, based on material collected by the writer as a member of the Smithsonian Biological Survey of the Panama Canal Zone during the first half of the year 1911. ... (PDF contains 13 pages)

Measurements of resistance and reactance in fish with the use of bioelectrical impedance analysis: sources of error

New technologies can be riddled with unforeseen sources of error, jeopardizing the validity and application of their advancement. Bioelectrical impedance analysis (BIA) is a new technology in fisheries research that is capable of estimating proximate composition, condition, and energy content in fish quickly, cheaply, and (after calibration) without the need to sacrifice fish. Before BIA can be widely accepted in fisheries science, it is necessary to identify sources of error and determine a means to minimize potential errors with this analysis. We conducted controlled laboratory experiments to identify sources of errors within BIA measurements. We concluded that electrode needle location, procedure deviations, user experience, time after death, and temperature can affect resistance and reactance measurements. Sensitivity analyses showed that errors in predictive estimates of composition can be large (>50%) when these errors are experienced. Adherence to a strict protocol can help avoid these sources of error and provide BIA estimates that are both accurate and precise in a field or laboratory setting.

On-going generation of multiple forms of HIV-1 intersubtype recombinants in the Yunnan Province of China

Objectives: To investigate the molecular epidemiology of HIV in China's Yunnan Province, where the initial HIV-1 outbreak among injecting drug users (IDU) occurred in 1989, and to analyse the genesis and interrelationship of the epidemic with that in surrounding areas. Design: A molecular epidemiological investigation was conducted among IDU in three prefectures in Yunnan Province, including Wenshan (east), Honghe (southeast) and Dehong (west). Methods: Thirty-nine specimens were collected from consenting IDU in 2000-2001. The nucleotide sequences of 2.6 kb gag-RT and 340 base pair (bp) env (C2/V3) regions were determined. Phylogenetic tree and recombination breakpoint analyses were performed. Results: The circulating recombinant form (CRF), CRF08_BC, predominated in east Yunnan near Guangxi Province (89% in Wenshan and 81% in Honghe), whereas it was not detected in Dehong(0/14) in the west. In contrast, 71% (10/14) of the Dehong isolates were unique recombinant forms (URF), mostly between subtypes B' (Thailand variant of subtype B) and C, with distinct profiles of recombination breakpoints. The subtype B' accounts for the remaining 29% (4/14) of Dehong isolates. Interestingly, two Honghe isolates (2/16) shared some of the precise B'/C recombination breakpoints with CRF07_BC. Conclusion: New recombinant strains are arising continually in west Yunnan near the Myanmar border. Some appeared to be secondary recombinants derived from CRF07_BC that had further recombined with other strains. The uneven distribution of subtypes, CRF and URF, suggests the presence of independent transmission networks and clusters among IDU in Yunnan. (C) 2002 Lippincott Williams Wilkins.

A new structure of In-based ohmic contacts to n-type GaAs

Electrical, structural and reaction characteristics of In-based ohmic contacts to n-GaAs were studied. Attempts were made to form a low-band-gap interfacial phase of InGaAs to reduce the barrier height at the metal/semiconductor junction, thus yielding low-resistance, highly reliable contacts. The contacts were fabricated by e-beam sputtering Ni, NiIn and Ge targets on VPE-grown n(+)-GaAs film (approximate to 1 mu m, 2 x 10(18) cm(-3)) in ultrahigh vacuum as the structure of Ni(200 Angstrom)/NiIn(100 Angstrom)/Ge(40 Angstrom)/n(+)-GaAs/SI-GaAs, followed by rapid thermal annealing at various temperatures (500-900 degrees C). In this structure, a very thin layer of Ge was employed to play the role of heavily doping donors and diffusion limiters between In and the GaAs substrate. Indium was deposited by sputtering NiIn alloy instead of pure In in order to ensure In atoms to be distributed uniformly in the substrate; nickel was chosen to consume the excess indium and form a high-temperature alloy of Ni3In. The lowest specific contact resistivity (rho(c)) of (1.5 +/- 0.5)x 10(-6) cm(2) measured by the Transmission Line Method (TLM) was obtained after annealing at 700 degrees C for 10 s. Auger sputtering depth profile and Transmission Electron Microscopy (TEM) were used to analyze the interfacial microstructure. By correlating the interfacial microstructure to the electronical properties, InxGa1-xAs phases with a large fractional area grown epitaxially on GaAs were found to be essential for reduction of the contact resistance.

Metagentiana, a new genus of Gentianaceae

The evidence from cross morphology, floral anatomy, chromosomes, palynology, and embryology all indicates that sect. Stenogyne is discordant within the genus Gentiana and is as distinct from the other sections of Gentiana as are other genera, such as Tripterospermum and Crawfurdia. In light of these characters, sect. Stenogyne is removed from Gentiana and given generic rank as the new genus Metagentiana. It is more related to Tripterospermum and Crawfurdia than to Gentiana, though it is more primitive than the first two genera. Together with Tripterospermum and Crawfurdia the new genus forms a monophyletic group, which is the sister group to the genus Gentiana. Fourteen new combinations required at specific rank are proposed.

Typical reggaeton in Morocco: forms of female political affirmation throught dance

Through an ethnographic account, this text analyses how social dance may become a discourse involving the cultural affirmation of a subordinate group. It describes how a group of girls faced with a complex of outlooks that construed them as Moroccan, Muslim or unattractive —or as objects of education and intervention— responded by affirming their own culture with an unanticipated corporal discourse. The way in which looking construes bodies is explored through metaphors: a hand that touches, a chisel that sculpts, a whip that lashes and a cobweb that controls and traps bodies. Owing to this political dimension of dance, workshops can also be an oppressive and silencing tool; to prevent this, the article concludes with a series of recommendations to implement dance in social intervention processes.

Enhanced cAMP generation and insulin-releasing potency of two novel Tyr(1)-modified enzyme-resistant forms of glucose-dependent insulinotropic polypeptide is associated with significant antihyperglycaemic activity in spontaneous obesity-diabetes

Glucose-dependent insulinotropic polypeptide (GIP) is an important incretin hormone, which potentiates glucose-induced insulin secretion. Antihyperglycaemic actions of GIP provide significant potential in Type 11 diabetes therapy. However, inactivation of GIP by the enzyme dipeptidyl peptidase IV (DPP IV) and its consequent short circulating half-life limit its therapeutic use. Therefore two novel Tyr(1)-Modified analogues of GIP, N-Fmoc-GIP (where Fmoc is 9-fluorenylmethoxycarbonyl) and N-palmitate-GIP, were synthesized and tested for metabolic stability and biological activity. Both GIP analogues were resistant to degradation by DPP IV and human plasma. In Chinese hamster lung (CHL) cells expressing the cloned human GIP receptor, both analogues exhibited a 2-fold increase in cAMP-generating potency compared with native GIP (EC50 values of 9.4, 10.0 and 18.2 nM respectively). Using clonal BRIN-BD11 cells, both analogues demonstrated strong insulinotropic activity compared with native GIP (P <0.01 to P <0.001). In obese diabetic (ob/ob) mice, administration of N-Fmoc-GIP or N-palmitate-GIP (25 nmol/kg) together with glucose (18 mmol/kg) significantly reduced the peak 15 min glucose excursion (1.4- and 1.5-fold respectively; P <0.05 to P <0.01) compared with glucose alone. The area under the curve (AUC) for glucose was significantly lower after administration of either analogue compared with glucose administered alone or in combination with native GIP (1.5-fold; P <0.05). This was associated with a significantly greater AUC for insulin (2.1-fold; P <0.001) for both analogues compared with native GIP. A similar pattern of in vivo responsiveness was evident in lean control mice. These data indicate that novel N-terminal Tyr(1) modification of GIP with an Fmoc or palmitate group confers resistance to degradation by DPP IV in plasma, which is reflected by increased in vitro potency and greater insulinotropic and antihyperglycaemic activities in an animal model of Type 11 diabetes mellitus.