Differences in Transporters Rather than Drug Targets Are the Principal Determinants of the Different Innate Sensitivities of Trypanosoma congolense and Trypanozoon Subgenus Trypanosomes to Diamidines and Melaminophenyl Arsenicals

The animal trypanosomiases are infections in a wide range of (domesticated) animals with any species of African trypanosome, such as Trypanosoma brucei, T. evansi, T. congolense, T. equiperdum and T. vivax. Symptoms differ between host and infective species and stage of infection and are treated with a small set of decades-old trypanocides. A complication is that not all trypanosome species are equally sensitive to all drugs and the reasons are at best partially understood. Here, we investigate whether drug transporters, mostly identified in T. b. brucei, determine the different drug sensitivities. We report that homologues of the aminopurine transporter TbAT1 and the aquaporin TbAQP2 are absent in T. congolense, while their introduction greatly sensitises this species to diamidine (pentamidine, diminazene) and melaminophenyl (melarsomine) drugs. Accumulation of these drugs in the transgenic lines was much more rapid. T. congolense is also inherently less sensitive to suramin than T. brucei, despite accumulating it faster. Expression of a proposed suramin transporter, located in T. brucei lysosomes, in T. congolense, did not alter its suramin sensitivity. We conclude that for several of the most important classes of trypanocides the presence of specific transporters, rather than drug targets, is the determining factor of drug efficacy.     

MicroRNA 148a Suppresses Tuberculous Fibrosis by Targeting NOX4 and POLDIP2

Extracellular matrix production by pleural mesothelial cells in response to Mycobacterium tuberculosis contributes to tuberculous fibrosis. NOX4 is involved in the pathogenesis of tuberculous fibrosis. In this study, we evaluated whether NOX4 gene-targeting microRNAs showed protective effects in tuberculosis fibrosis. TargetScan prediction software was used to identify candidate microRNAs that bind the 3′ UTRs of NOX4, and microRNA-148a (miR-148a) was selected as the best miRNA candidate. A repressed and forced expression assay in Met5A cells was performed to investigate the causal relationship between miR-148a and NOX4. The role of miR-148a in tuberculous pleural fibrosis was studied using a murine model of Mycobacterium bovis bacillus Calmette–Guérin (BCG) pleural infection. Heat-killed M. tuberculosis (HKMT) induces NOX4 and POLDIP2 expression. We demonstrated the inhibitory effect of miR-148a on NOX4 and POLDIP2 expression. The increased expression of miR-148a suppressed HKMT-induced collagen-1A synthesis in PMC cells. In the BCG pleurisy model, miR-148a significantly reduced fibrogenesis and epithelial mesenchymal transition. High levels of miR-148a in tuberculous pleural effusion can be interpreted as a self-limiting homeostatic response. Our data indicate that miR-148a may protect against tuberculous pleural fibrosis by regulating NOX4 and POLDIP2.     

A Toxoplasma gondii Oxopurine Transporter Binds Nucleobases and Nucleosides Using Different Binding Modes

Toxoplasma gondii is unable to synthesize purines de novo, instead salvages them from its environment, inside the host cell, for which they need high affinity carriers. Here, we report the expression of a T. gondii Equilibrative Nucleoside Transporter, Tg244440, in a Trypanosoma brucei strain from which nucleobase transporters have been deleted. Tg244440 transported hypoxanthine and guanine with similar affinity (K_m ~1 µM), while inosine and guanosine displayed K_i values of 4.05 and 3.30 µM, respectively. Low affinity was observed for adenosine, adenine, and pyrimidines, classifying Tg244440 as a high affinity oxopurine transporter. Purine analogues were used to probe the substrate-transporter binding interactions, culminating in quantitative models showing different binding modes for oxopurine bases, oxopurine nucleosides, and adenosine. Hypoxanthine and guanine interacted through protonated N1 and N9, and through unprotonated N3 and N7 of the purine ring, whereas inosine and guanosine mostly employed the ribose hydroxy groups for binding, in addition to N1H of the nucleobase. Conversely, the ribose moiety of adenosine barely made any contribution to binding. Tg244440 is the first gene identified to encode a high affinity oxopurine transporter in T. gondii and, to the best of our knowledge, the first purine transporter to employ different binding modes for nucleosides and nucleobases.     

Spatial concentration of manufacturing firms in China

Applying the methodology developed by Duranton and Overman, we analyse localization and dispersion of firms in China. Using a unique and detailed dataset on manufacturing firms in China, we are able to follow the changes in location patterns of firms between 2002 and 2008. Our analysis shows that firms in China are more localized than in the UK or Japan. Localization is comparable to that in the US, and takes place at relatively small scales that are consistent with the size of Chinese cities. Localization increases rapidly, even in the relative short period between 2002 and 2008, especially new entrants localize. Private firms, firms from Hong‐Kong, Macao and Taiwan, and foreign firms are more localized than state‐owned firms. Our findings are consistent with the notion that China is increasingly liberalizing its economy, enabling (profit seeking) manufacturing firms to benefit from agglomeration economies.     

Adsorption of lutein from soybean oil on silicic acid I. Isotherms

Adsorption of lutein from crude soybean oil miscella on dispersed silicic acid resulted in three different Freundlich type isotherms, depending on the amount of adsorbent used. Normally, changing the amount of adsorbent gives a new point on the same isotherm. Further investigation of the lutein adsorption revealed that addition of 1% isopropanol to the hexane solvent and deactivation of the silica acid with water decreased lutein adsorption. Purification of the lutein resulted in increased adsorption, indicating that triglyceride was competing for adsorption sites. The triglyceride competition was confirmed by calculating an adsorption isotherm for triglyceride from crude soybean oil miscella. Use of silicic acid as a column rather than dispersed yielded multiple isotherms for lutein adsorption based on the amount of adsorbent used. Also, lutein adsorption on columns was concentrated at the entrance to the column. Experiments showed that lutein adsorption was not due to partitioning between bound triglyceride and hexane solvent.     

Study of the reaction of cokes with KOH under nitrogen

Five cokes of increasing content of anisotropic carbon were prepared. Polished surfaces of these cokes were characterized by optical microscopy in terms of components of optical texture. These surfaces were reacted with KOH at 873, 1073 and 1273 K in an inert atmosphere for 2 h and the resultant topography monitored by scanning electron microscopy (SEM). The extent of potassium take-up by coke particles was measured and the diffusion of potassium was detected by EDAX. Microstrength testing was made on the cokes before and after reaction with the alkali. Coke reactivity measurements were obtained for untreated and treated cokes. Results indicate that in an inert atmosphere the alkali reacts preferentially on the prismatic edges of anisotropic carbon and that the rates of reaction increase with increasing temperature. Potassium is able to diffuse into the interior of the more anisotropic coke particles and this casues weakening of the coke. The reactivity measurements indicate that for the more anisotropic cokes the effect of potassium as a catalyst in the solution-loss reaction is more pronounced than for the least anisotropic coke. These conclusions suggest that metallurgical coke in the blast furnace in the presence of alkali materials can lose strength by direct reaction over and above considerations of gasification processes.     

Substituent distribution within cross-linked and hydroxypropylated sweet potato starch and potato starch

Revealing the substituents distribution within starch can help to understand the changes of starch properties after modification. The distribution of substituents over cross-linked and hydroxypropylated sweet potato starch was investigated and compared with modified potato starch. The starches were cross-linked with sodium trimetaphosphate and/or hydroxypropylated with propylene oxide. The native and modified starches were gelatinized and hydrolysed by pullulanase, β-amylase, α-amylase and a combination of pullulanase, α-amylase and amyloglucosidase. The hydrolysates were analysed by HPSEC, HPAEC and MALDI-TOF mass spectrometry. Cross-linking had only a slight effect on the enzymatic hydrolysis, where hydroxypropylation evidently limited the enzymatic hydrolysis. The results obtained suggest that the hydroxypropyl substituents are not distributed regularly over the starch chains. Although the average substitution was around 2 hydroxypropyl groups per 10 glucose units, in the enzyme digests of hydroxypropylated starches, oligomer fragments of 10–15 glucose units, carrying 5–8 hydroxypropyl groups, were identified. It is hypothesised that higher levels of substituents are present in the amorphous regions and periphery of clusters of starch granules. This is the first time that the location of hydroxypropyl groups within sweet potato starch has been examined in this detail. Despite significant differences in granule architecture between starches from potato and sweet potato, similar patterns of hydroxypropylation have been found.     

Automatic Absorbance Calibration Routine for a Computerized Uv-Visible Rapid Scanning Spectrophotometer

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A Fortran IV computer program is used to calculate absorbance at varying wavelengths in a UV-visible rapid scanning spectrophotometer. Using the program eliminates the necessity of calibrating the absorbance axis in the usual manner using standard solutions or filters and plotting calibration curves, because the voltage output of the computer is automatically identical to the actual absorbance of a sample.     

Determination of the distribution of air and water in porous media by electrical impedance tomography and magneto-electrical imaging

Monitoring the distribution of water content is essential for understanding hydrological processes in the lithosphere and the pedosphere. The movement of water in unsaturated rock formations and in the vadose zone is influenced by different processes (mainly infiltration, evaporation, percolation and capillary flow) which may be rate determining depending on the actual conditions. The interdependence of these processes also strongly influences the transport and distribution of solutes in the pore space. In order to gain a better understanding of the movement and distribution of water in unsaturated media, systematic investigations with non-invasive or minimal invasive methods appear to be most suitable. Studies on the distribution of electrical conductivity can improve risk analysis concerning waste disposals in general and nuclear waste repositories in particular. Induced polarization and magnetic flux density determined with two highly sensitive accessories yield additional information and may allow for better discrimination of coupled flow processes. Electrical impedance tomography (EIT) with 20 current injection and 48 voltage electrodes was used here to monitor the evaporation of tap water from a container filled with sand under laboratory conditions at 20 °C. The results are compared with data obtained by determining spectral induced polarization (SIP) of sand during desaturation in a multi-step outflow equipment. Infiltration processes and evaporation from sand saturated with 0.01 M CaCl₂ were determined by magneto-electrical resistivity imaging technique (MERIT). The results were obtained from a long-term experiment under controlled conditions.