Trapped lipopolysaccharide and LptD intermediates reveal lipopolysaccharide translocation steps across the Escherichia coli outer membrane

Lipopolysaccharide (LPS) is a main component of the outer membrane of Gram-negative bacteria, which is essential for the vitality of most Gram-negative bacteria and plays a critical role for drug resistance. LptD/E complex forms a N-terminal LPS transport slide, a hydrophobic intramembrane hole and the hydrophilic channel of the barrel, for LPS transport, lipid A insertion and core oligosaccharide and O-antigen polysaccharide translocation, respectively. However, there is no direct evidence to confirm that LptD/E transports LPS from the periplasm to the external leaflet of the outer membrane. By replacing LptD residues with an unnatural amino acid p-benzoyl-L-phenyalanine (pBPA) and UV-photo-cross-linking in E.coli, the translocon and LPS intermediates were obtained at the N-terminal domain, the intramembrane hole, the lumenal gate, the lumen of LptD channel, and the extracellular loop 1 and 4, providing the first direct evidence and “snapshots” to reveal LPS translocation steps across the outer membrane

Amebiasis in HIV-1-Infected Japanese Men: Clinical Features and Response to Therapy

Invasive amebic diseases caused by Entamoeba histolytica are increasing among men who have sex with men and co-infection of ameba and HIV-1 is an emerging problem in developed East Asian countries. To characterize the clinical and epidemiological features of invasive amebiasis in HIV-1 patients, the medical records of 170 co-infected cases were analyzed retrospectively, and E. histolytica genotype was assayed in 14 cases. In this series of HIV-1-infected patients, clinical presentation of invasive amebiasis was similar to that described in the normal host. High fever, leukocytosis and high CRP were associated with extraluminal amebic diseases. Two cases died from amebic colitis (resulting in intestinal perforation in one and gastrointestinal bleeding in one), and three cases died from causes unrelated to amebiasis. Treatment with metronidazole or tinidazole was successful in the other 165 cases. Luminal treatment was provided to 83 patients following metronidazole or tinidazole treatment. However, amebiasis recurred in 6 of these, a frequency similar to that seen in patients who did not receive luminal treatment. Recurrence was more frequent in HCV-antibody positive individuals and those who acquired syphilis during the follow-up period. Various genotypes of E. histolytica were identified in 14 patients but there was no correlation between genotype and clinical features. The outcome of metronidazole and tinidazole treatment of uncomplicated amebiasis was excellent even in HIV-1-infected individuals. Luminal treatment following metronidazole or tinidazole treatment does not reduce recurrence of amebiasis in high risk populations probably due to amebic re-infection

Novel insights into host-fungal pathogen interactions derived from live-cell imaging

Acknowledgments The authors acknowledge funding from the Wellcome Trust (080088, 086827, 075470 and 099215) including a Wellcome Trust Strategic Award for Medical Mycology and Fungal Immunology 097377 and FP7-2007–2013 grant agreement HEALTH-F2-2010-260338–ALLFUN to NARG.Peer reviewedPublisher PD

Analysis of parcel-based image classification methods for monitoring the activities of the Land Bank of Galicia (Spain)

[EN] The abandonment of agricultural plots entails a low economic productivity of the land and a higher vulnerability to wildfires and degradation of affected areas. In this sense, the local government of Galicia is promoting new methodologies based on high-resolution images in order to classify the territory in basic and generic land uses. This procedure will be used to control the sustainable management of plots belonging to the Land Bank. This paper presents an application study for maintaining and updating land use/land cover geospatial databases using parcel-oriented classification. The test is performed over two geographic areas of Galicia, in the northwest of Spain. In this region, forest and shrublands in mountain environments are very heterogeneous with many private unproductive plots, some of which are in a high state of abandonment. The dataset is made of high spatial resolution multispectral imagery, cadastral cartography employed to define the image objects (plots), and field samples used to define evaluation and training samples. A set of descriptive features is computed quantifying different properties of the objects, i.e. spectral, texture, structural, and geometrical. Additionally, the effect on the classification and updating processes of the historical land use as a descriptive feature is tested. Three different classification methodologies are analyzed: linear discriminant analysis, decision trees, and support vector machine. The overall accuracies of the classifications obtained are always above 90 % and support vector machine method is proved to provide the best performance. Forest and shrublands areas are especially undefined, so the discrimination between these two classes is low. The results enable to conclude that the use of automatic parcel-oriented classification techniques for updating tasks of land use/land cover geospatial databases, is effective in the areas tested, particularly when broad and well defined classes are required.The authors appreciate the collaboration and support provided by Xunta de Galicia, Sociedade para o Desenvolvemento Comarcal de Galícia, and Banco de Terras de Galicia. The financial support provided by the Spanish Ministerio de Ciencia e Innovación in the framework of the projects CGL2010-19591/BTE and CGL2009-14220 is also acknowledged.Hermosilla, T.; Díaz Manso, J.; Ruiz Fernández, LÁ.; Recio Recio, JA.; Fernández-Sarría, A.; Ferradáns Nogueira, P. (2012). Analysis of parcel-based image classification methods for monitoring the activities of the Land Bank of Galicia (Spain). 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Int Arch Photogramm Remote Sens Spat Inf Sci 38:217–222Perveen F, Nagasawa R, Ali S, Husnain (2008) Evaluation of ASTER spectral bands for agricultural land cover mapping using pixel-based and object-based classification approaches. Int Arch Photogramm Remote Sens Spat Inf Sci 37(4-C1)Petit CC, Lambin EF (2002) Impact of data integration technique on historical land-use/land-cover change: comparing historical maps with remote sensing data in the Belgian Ardennes. Landsc Ecol 17:117–132Quinlan JR (1993) C4.5: Programs for machine learning. Kaufmann, San FranciscoRabe A, van der Linden S, Hostert P (2010) imageSVM, Version 2.1. www.hu-geomatics.deRecio JA, Hermosilla T, Ruiz LA, Fernández-Sarría A (2011) Historical land use as a feature for image classification. Photogramm Eng Remote Sens 77(4):377–387Ruiz LA, Fernández-Sarría A, Recio JA (2004) Texture feature extraction for classification of remote sensing data using wavelet decomposition: a comparative study. 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Postoperative IOP prophylaxis practice following uncomplicated cataract surgery: a UK-wide consultant survey

BACKGROUND: In order to minimise postoperative intraocular pressure (IOP) rise, after routine uncomplicated cataract surgery, prophylaxis may be adopted. Currently, there are no specific guidelines in this regard resulting in wide variation in practice across the UK. We sought to document these variations through a questionnaire survey. METHODS: A questionnaire was sent to all consultant ophthalmic surgeons in the UK. RESULTS: 62.6% of surgeons did not use any IOP lowering agents. 37.4% surgeons routinely prescribed some form of medication. The majority (86.8%) used oral diamox. 20.6% of surgeons said they based their practice on evidence, 43.3% on personal experience, and 17.6% on unit policy. Surprisingly, among the two groups of surgeons (those who gave routine prophylaxis, and those who did not) the percentages of surgeons quoting personal experience, unit policy, or presence of evidence was strikingly similar. The timing of the first postoperative IOP check varied from the same day to beyond 2 weeks. Only 20.2% of surgeons had ever seen an adverse event related to IOP rise; this complication is thus very rare. CONCLUSION: This survey highlights a wide variation in the practice and postoperative management of phacoemulsification cataract surgery. What is very striking is that there is a similar proportion of surgeons in the diametrically opposite groups (those who give or do not give routine IOP lowering prophylaxis) who believe that there practice is evidence based. The merits of this study suggests that consideration must be given to drafting a uniform guideline in this area of practice

Characterization of mercury bioremediation by transgenic bacteria expressing metallothionein and polyphosphate kinase

<p>Abstract</p> <p>Background</p> <p>The use of transgenic bacteria has been proposed as a suitable alternative for mercury remediation. Ideally, mercury would be sequestered by metal-scavenging agents inside transgenic bacteria for subsequent retrieval. So far, this approach has produced limited protection and accumulation. We report here the development of a transgenic system that effectively expresses metallothionein (<it>mt-1</it>) and polyphosphate kinase (<it>ppk</it>) genes in bacteria in order to provide high mercury resistance and accumulation.</p> <p>Results</p> <p>In this study, bacterial transformation with transcriptional and translational enhanced vectors designed for the expression of metallothionein and polyphosphate kinase provided high transgene transcript levels independent of the gene being expressed. Expression of polyphosphate kinase and metallothionein in transgenic bacteria provided high resistance to mercury, up to 80 μM and 120 μM, respectively. Here we show for the first time that metallothionein can be efficiently expressed in bacteria without being fused to a carrier protein to enhance mercury bioremediation. Cold vapor atomic absorption spectrometry analyzes revealed that the <it>mt-1 </it>transgenic bacteria accumulated up to 100.2 ± 17.6 μM of mercury from media containing 120 μM Hg. The extent of mercury remediation was such that the contaminated media remediated by the <it>mt-1 </it>transgenic bacteria supported the growth of untransformed bacteria. Cell aggregation, precipitation and color changes were visually observed in <it>mt-1 </it>and <it>ppk </it>transgenic bacteria when these cells were grown in high mercury concentrations.</p> <p>Conclusion</p> <p>The transgenic bacterial system described in this study presents a viable technology for mercury bioremediation from liquid matrices because it provides high mercury resistance and accumulation while inhibiting elemental mercury volatilization. This is the first report that shows that metallothionein expression provides mercury resistance and accumulation in recombinant bacteria. The high accumulation of mercury in the transgenic cells could present the possibility of retrieving the accumulated mercury for further industrial applications.</p

Correcting non cephalic presentation with moxibustion: study protocol for a multi-centre randomised controlled trial in general practice

<p>Abstract</p> <p>Background</p> <p>Non cephalic presentation in childbirth involves various risks to both the mother and the foetus. The incidence in Spain is 3.8% of all full-term pregnancies. The most common technique used to end the gestation in cases of non cephalic presentation is that of caesarian section, and although it provokes a lower rate of morbi-mortality than does vaginal delivery in such situations, there remains the possibility of traumatic injury to the foetal head and neck, while maternal morbidity is also increased. The application of heat (moxibustion) to an acupuncture point, in order to correct non cephalic presentation, has been practised in China since ancient times, but as yet there is insufficient evidence of its real effectiveness.</p> <p>Methods/Design</p> <p>The experimental design consists of a multi-centre randomised controlled trial with three parallel arms, used to compare real moxibustion, sham moxibustion and the natural course of events, among pregnant women with a non cephalic presentation and a gestational duration of 33–35 weeks (estimated by echography). The participants in the trial will be blinded to both interventions. The results obtained will be analyzed by professionals, blinded with respect to the allocation to the different types of intervention. In addition, we intend to carry out a economic analysis.</p> <p>Discussion</p> <p>This trial will contribute to the development of evidence concerning moxibustion in the correction of non cephalic presentations. The primary outcome variable is the proportion of cephalic presentations at term. As secondary outcomes, we will evaluate the proportion of cephalic presentations at week 38 of gestation, determined by echography, together with the safety of the technique, the specificity of moxibustion and the control of the blinding process.</p> <p>This study has been funded by the Health Ministry of the Andalusian Regional Government.</p> <p>Trial registration</p> <p>Current Controlled Trials ISRCTN10634508.</p

An unusual presentation of anetoderma: a case report

BACKGROUND: Anetoderma is a benign condition with focal loss of dermal elastic tissue resulting in localized areas of flaccid or herniated saclike skin. Currently, anetoderma is classified as either primary (idiopathic), or secondary anetoderma (which is associated with a variety of skin conditions, penicillamine use, or neonatal prematurity). Lesions appear on the upper arms, trunk, and thighs. CASE PRESENTATION: We report a 14-year-old boy, which was noticed to have had multiple, white, non-pruritic areas on the acral sites of upper and lower extremities for two years. In physical examination, the patient had normal mental development. Skin lesions consisted of scattered, white to skin-colored papules, less than 1 cm in diameter, and with central protrusion, with distribution on dorsal part of the index finger, forearms, distal portion of thighs and calves. Lesions were detected neither on the trunk nor the proximal areas of extremities. There are no sensory changes associated with the lesions. Otherwise, his general health was good. He did not have any medication consumption history. Family history was negative. Laboratory examinations were within normal limits. Skin biopsy from one of his lesions was done, that confirmed the diagnosis of anetoderma. CONCLUSIONS: In summary, we report a case of anetoderma on unusual sites of the skin. We could not find similar reports of anetoderma developing on distal extremities without involvement of the upper trunk and proximal arms, in the medical literature

Immunological predictors of CD4+ T cell decline in antiretroviral treatment interruptions

<p>Abstract</p> <p>Background</p> <p>The common response to stopping anti-HIV treatment is an increase of HIV-RNA load and decrease in CD4⁺, but not all the patients have similar responses to this therapeutic strategy. The aim was to identify predictive markers of CD4⁺cell count declines to < 350/μL in CD4-guided antiretroviral treatment interruptions.</p> <p>Methods</p> <p>27 HIV-infected patients participated in a prospective multicenter study in with a 24 month follow-up. Patients on stable highly active antiretroviral therapy (HAART), with CD4⁺count > 600/μL, and HIV-RNA < 50 copies/ml for at least 6 months were offered the option to discontinue antiretroviral therapy. The main outcome was a decline in CD4⁺cell count to < 350/μL.</p> <p>Results</p> <p>After 24 months of follow-up, 16 of 27 (59%) patients (who discontinued therapy) experienced declines in CD4⁺cell count to < 350/μL. Patients with a CD4⁺nadir of < 200 cells/μL had a greater risk of restarting therapy during the follow-up (RR (CI95%): 3.37 (1.07; 10.36)). Interestingly, lymphoproliferative responses to <it>Mycobacterium tuberculosis </it>purified protein derivative (PPD) below 10000 c.p.m. at baseline (4.77 (1.07; 21.12)), IL-4 production above 100 pg/mL at baseline (5.95 (1.76; 20.07)) in PBMC cultured with PPD, and increased IL-4 production of PBMC with p24 antigen at baseline (1.25 (1.01; 1.55)) were associated to declines in CD4⁺cell count to < 350/μL.</p> <p>Conclusion</p> <p>Both the number (CD4⁺nadir) and the functional activity of CD4⁺(lymphoproliferative response to PPD) predict the CD4⁺decrease associated with discontinuation of ART in patients with controlled viremia.</p