Treatment of tuberculosis in a region with high drug resistance: outcomes, drug resistance amplification and re-infection

Introduction

Emerging antituberculosis drug resistance is a serious threat for tuberculosis (TB) control, especially in Eastern European countries.

Methods

We combined drug susceptibility results and molecular strain typing data with treatment outcome reports to assess the influence of drug resistance on TB treatment outcomes in a prospective cohort of patients from Abkhazia (Georgia). Patients received individualized treatment regimens based on drug susceptibility testing (DST) results. Definitions for antituberculosis drug resistance and treatment outcomes were in line with current WHO recommendations. First and second line DST, and molecular typing were performed in a supranational laboratory for Mycobacterium tuberculosis (MTB) strains from consecutive sputum smear-positive TB patients at baseline and during treatment.

Results

At baseline, MTB strains were fully drug-susceptible in 189/326 (58.0%) of patients. Resistance to at least H or R (PDR-TB) and multidrug-resistance (MDR-TB) were found in 69/326 (21.2%) and 68/326 (20.9%) of strains, respectively. Three MDR-TB strains were also extensively resistant (XDR-TB). During treatment, 3/189 (1.6%) fully susceptible patients at baseline were re-infected with a MDR-TB strain and 2/58 (3.4%) PDR-TB patients became MDR-TB due to resistance amplification. 5/47 (10.6%) MDR- patients became XDR-TB during treatment. Treatment success was observed in 161/189 (85.2%), 54/69 (78.3%) and 22/68 (32.3%) of patients with fully drug susceptible, PDR- and MDR-TB, respectively. Development of ofloxacin resistance was significantly associated with a negative treatment outcome.

Conclusion

In Abkhazia, a region with high prevalence of drug resistant TB, the use of individualized MDR-TB treatment regimens resulted in poor treatment outcomes and XDR-TB amplification. Nosocomial transmission of MDR-TB emphasizes the importance of infection control in hospitals.     

Use of the paired samples (cerebrospinal fluid and serum) in immunodiagnostic of active and inactive human neurocysticercosis

Paired samples of cerebrospinal fluid (CSF) and serum of 30 patients--10 with active, 10 with inactive neurocysticercosis (NCC), and 10 control subjects--were evaluated by enzyme-linked immunosorbent assay (ELISA) using two Taenia crassiceps metacestode extracts as antigen in order to detect IgG antibodies. In active NCC, high levels of IgG were detected (p < 0.05). The CSF samples showed 80% (CI 72-88) of reactivity in the saline extract (S) and 90% (CI 84-95) in sodium dodecyl sulphate (SDS) and the serum samples were reactive in 90% (CI 84-95) and 100% (CI 98-100) in the S and SDS antigenic extracts, respectively. The use of the paired samples of CSF and serum in active NCC showed equivalent results suggesting that the serum samples could be used as a screening in those patients whose CSF puncture is counter-indicated.     

Circular dichroism of the peripheral chlorophylls in photosystem II reaction centers revealed by electrochemical oxidation

Visible absorption spectra and circular dichroism (CD) of the red absorption band of isolated photosystem II reaction centers were measured at room temperature during progressive bleaching by electrochemical oxidation, in comparison with aerobic photochemical destruction, and with anaerobic photooxidation in the presence of the artificial electron acceptor silicomolybdate. Initially, selective bleaching of peripheral chlorophylls absorbing at 672 nm was obtained by electrochemical oxidation at +0.9 V, whereas little selectivity was observed at higher potentials. Illumination in the presence of silicomolybdate did not cause a bleaching but a spectral broadening of the 672-nm band was observed, apparently in response to the oxidation of carotene. The 672-nm absorption band is shown to exhibit a positive CD, which accounts for the 674-nm shoulder in CD spectra at low temperature. The origin of this CD is discussed in view of the observation that all CD disappears with the 680-nm absorption band during aerobic photodestruction.     

Geldanamycins trigger a novel Ron degradative pathway, hampering oncogenic signaling

Ron, the tyrosine kinase receptor for macrophage-stimulating protein is responsible for proliferation and migration of cells from different tissues. Ron can acquire oncogenic potential by single point mutations in the kinase domain, and dysregulated Ron signaling has been involved in the development of different human cancers. We have previously shown that ligand-activated Ron recruits the negative regulator c-Cbl, which mediates its ubiquitylation and degradation. Here we report that Ron is ubiquitylated also by the U-box E3 ligase C-terminal Hsc70-interacting protein (CHIP), recruited via chaperone intermediates Hsp90 and Hsc70. Gene silencing shows that CHIP activity is necessary to mediate Ron degradation upon cell treatment with Hsp90 inhibitors geldanamycins. The oncogenic Ron(M1254T) receptor escapes from c-Cbl negative regulation but retains a strong association with CHIP. This constitutively active mutant of Ron displays increased sensitivity to geldanamycins, enhanced physical interaction with Hsp90, and more rapid degradation rate. Cell growth and migration, as well as the transforming potential evoked by Ron(M1254T), are abrogated upon Hsp90 inhibition. These data highlight a novel mechanism for Ron degradation and propose Hsp90 antagonists like geldanamycins as suitable pharmacological agents for therapy of cancers where altered Ron signaling is involved.     

The influence of mandibular implant-retained overdentures in masticatory efficiency

doi: 10.1111/j.1741‐2358.2011.00539.x The influence of mandibular implant‐retained overdentures in masticatory efficiency Objective: To evaluate the masticatory efficiency of patients rehabilitated with conventional dentures (CDs) or implant‐retained mandibular overdentures. Background: Despite the evident benefits of implants on mastication as assessed by subjective patient‐based outcomes, the extent of implant overdenture treatment effect on food comminution is not well established. Materials and methods: A randomised clinical trial was carried out with 29 completely edentulous patients divided into two groups. The first group was rehabilitated with a mandibular overdenture retained by two splinted implants with bar‐clip system, while the second group was rehabilitated with a mandibular CD. Both groups also were rehabilitated with maxillary CDs. Masticatory efficiency and patient satisfaction were assessed 3 months after denture insertion. Masticatory efficiency was evaluated through the colorimetric method with the beads as the artificial test‐food. Comparisons for masticatory efficiency and patient satisfaction were performed using Student’s t‐test (α = 0.05). Results: No significant statistical difference was found for masticatory efficiency (p = 0.198). Patient overall satisfaction was significantly higher for the mandibular overdenture (p < 0.001). In addition, mandibular overdenture patients were significantly more satisfied with chewing experience (p < 0.05) and retention of the lower denture (p < 0.005). Conclusion: The results of this study suggest that mandibular overdenture significantly improves chewing experience, although limited effect on masticatory efficiency has been observed.     

Effects of grazing and invasive grasses on desert vertebrates in California

Much of California's San Joaquin Valley is a desert and, like portions of other North American deserts, is experiencing an ecological shift from being dominated by ephemeral native forbs, with widely spaced shrubs, to fire-prone non-native annual grasses. Small terrestrial vertebrates, many of which are adapted to open desert habitats, are declining. One hypothesis is that the invasive plants contribute to the decline by altering vegetative structure. Although cattle may have originally been a factor in the establishment of these non-native plants, their grazing may benefit the terrestrial vertebrates by maintaining an open structure, especially during average or wet winters when the exotic grasses grow especially dense. We experimentally tested the effect of cattle grazing on invasive plants and a community of small vertebrates at a site in the southwestern San Joaquin Desert. We established and monitored 4 treatment (grazed) and 4 control (ungrazed) plots from 1997 to 2006, and assessed the abundances of blunt-nosed leopard lizards (Gambelia sila), giant kangaroo rats (Dipodomys ingens), short-nosed kangaroo rats (Dipodomys nitratoides nitratoides), and San Joaquin antelope squirrels (Ammospermophilus nelsoni), all of which are listed as threatened or endangered by state or federal agencies. We also recorded abundances of the non-protected western whiptail lizards (Aspidoscelis tigris), side-blotched lizards (Uta stansburiana), San Joaquin pocket mice (Perognathus inornatus inornatus), and Heermann's kangaroo rats (Dipdomys heermanni). Based on repeated measures analysis of variance (ANOVA) and a 0.05 alpha level, only Heermann's kangaroo rats showed a treatment effect; they were more abundant on the control plots. However, this effect could be accounted for by the natural re-establishment of saltbush (Atriplex spp.) on part of the study site. Saltbush return also favored western whiptail lizards and San Joaquin antelope squirrels. A regression analysis indicated that populations of blunt-nosed leopard lizard and giant kangaroo rat increased significantly faster in grazed plots than the ungrazed controls, and abundances of 6 of 8 species were negatively correlated with increased residual dry matter. With relaxed alpha values to decrease Type II error, populations of blunt-nosed leopard lizards (500% greater), San Joaquin antelope squirrels (85% greater), and short-nosed kangaroo rats (73% greater) increased significantly on grazed plots over the course of the study compared to ungrazed plots. We did not find grazing to negatively affect abundance of any species we studied. When herbaceous cover is low during years of below average rainfall in deserts and other arid areas, grazing may not be necessary to maintain populations of small vertebrates. However, if cattle grazing is closely monitored in space and time to minimize adverse effects on the habitat, it could be an effective tool to control dense stands of non-native grasses and benefit native wildlife. © 2011 The Wildlife Society.     

Position of the third Na+ site in the aspartate transporter GltPh and the human glutamate transporter, EAAT1

Glutamate transport via the human excitatory amino acid transporters is coupled to the co-transport of three Na(+) ions, one H(+) and the counter-transport of one K(+) ion. Transport by an archaeal homologue of the human glutamate transporters, Glt(Ph), whose three dimensional structure is known is also coupled to three Na(+) ions but only two Na(+) ion binding sites have been observed in the crystal structure of Glt(Ph). In order to fully utilize the Glt(Ph) structure in functional studies of the human glutamate transporters, it is essential to understand the transport mechanism of Glt(Ph) and accurately determine the number and location of Na(+) ions coupled to transport. Several sites have been proposed for the binding of a third Na(+) ion from electrostatic calculations and molecular dynamics simulations. In this study, we have performed detailed free energy simulations for Glt(Ph) and reveal a new site for the third Na(+) ion involving the side chains of Threonine 92, Serine 93, Asparagine 310, Aspartate 312, and the backbone of Tyrosine 89. We have also studied the transport properties of alanine mutants of the coordinating residues Threonine 92 and Serine 93 in Glt(Ph), and the corresponding residues in a human glutamate transporter, EAAT1. The mutant transporters have reduced affinity for Na(+) compared to their wild type counterparts. These results confirm that Threonine 92 and Serine 93 are involved in the coordination of the third Na(+) ion in Glt(Ph) and EAAT1.