Structural Characterization of Phosphatidyl-myo-Inositol Mannosides from Mycobacterium bovis Bacillus Calmette Gúerin by Multiple-Stage Quadrupole Ion-Trap Mass Spectrometry with Electrospray Ionization. II. Monoacyl- and Diacyl-PIMs

The multiple-stage ion-trap mass spectrometric approaches towards to the structural characterization of the monoacyl-PIM (triacylated PIM) and the diacyl-PIM (tetracylated PIM), namely, the PIM (diacylated PIM) consisting of one or two additional fatty acid substituents attached to the glycoside, respectively, were described. While the assignment and confirmation of the fatty acid substituents on the glycerol backbone can be easily achieved by the methods described in the previous article, the identity of the glycoside moiety and its acylation state can be determined by the observation of a prominent acylglycoside ion arising from cleavage of the diacylglycerol moiety ([M − H − diacylglycerol]−) in the MS2-spectra of monoacyl-PIM and diacyl-PIM. The distinction of the fatty acid substituents on the 2-O-mannoside (i.e., R3CO2H) from that on the inositol (i.e., R4CO2H) is based on the findings that the MS3-spectrum of [M − H − diacylglycerol]− contains a prominent ion arising from further loss of the fatty acid at the 2-O-mannoside (i.e., the [M − H − diacylglycerol − R3CO2H]− ion), while the ion arising from loss of the fatty acid substituent at the inositol (i.e., the [M − H − diacylglycerol − R4CO2H]− ion) is of low abundance. The fatty acyl moiety on the inositol can also be identified by the product-ion spectrum from MS4 of the [M − H − diacylglycerol − R3CO2H]− ion, which gives rise to a prominent ion corresponding to loss of R4CO2H. An [M − H − acylmannose]− ion was also observed in the MS2-spectra and, thus, the identity of the fatty acid substituent attached to 2-O-mannoside can be confirmed. The combined information obtained from the multiple-stage product-ion spectra from MS2, MS3, and MS4 permit the assignment of the complex structures of monoacyl-PIMs and diacyl-PIMs in a mixture isolated from M. bovis Bacillus Calmette Guérin

Precipitation from Space: Advancing Earth System Science

Of the three primary sources of spatially contiguous precipitation observations (surface networks, ground-based radar, and satellite-based radar/radiometers), only the last is a viable source over ocean and much of the Earth's land. As recently as 15 years ago, users needing quantitative detail of precipitation on anything under a monthly time scale relied upon products derived from geostationary satellite thermal infrared (IR) indices. The Special Sensor Microwave Imager (SSMI) passive microwave (PMW) imagers originated in 1987 and continue today with the SSMI sounder (SSMIS) sensor. The fortunate longevity of the joint National Aeronautics and Space Administration (NASA) and Japan Aerospace Exploration Agency (JAXA) Tropical Rainfall Measuring Mission (TRMM) is providing the environmental science community a nearly unbroken data record (as of April 2012, over 14 years) of tropical and sub-tropical precipitation processes. TRMM was originally conceived in the mid-1980s as a climate mission with relatively modest goals, including monthly averaged precipitation. TRMM data were quickly exploited for model data assimilation and, beginning in 1999 with the availability of near real time data, for tropical cyclone warnings. To overcome the intermittently spaced revisit from these and other low Earth-orbiting satellites, many methods to merge PMW-based precipitation data and geostationary satellite observations have been developed, such as the TRMM Multisatellite Precipitation Product and the Climate Prediction Center (CPC) morphing method (CMORPH. The purpose of this article is not to provide a survey or assessment of these and other satellite-based precipitation datasets, which are well summarized in several recent articles. Rather, the intent is to demonstrate how the availability and continuity of satellite-based precipitation data records is transforming the ways that scientific and societal issues related to precipitation are addressed, in ways that would not be otherwise possible. These developments have taken place in parallel with the growth of an increasingly interconnected scientific environment. Scientists from different disciplines can easily interact with each other via information and materials they encounter online, and collaborate remotely without ever meeting each other in person. Likewise, these precipitation datasets are quickly and easily available via various data portals and are widely used. Within the framework of the NASA/JAXA Global Precipitation Measurement (GPM mission, these applications will become increasingly interconnected. We emphasize that precipitation observations by themselves provide an incomplete picture of the state of the atmosphere. For example, it is unlikely that a richer understanding of the global water cycle will be possible by standalone missions and algorithms, but must also involve some component of data, where model analyses of the physical state are constrained alongside multiple observations (e.g., precipitation, evaporation, radiation). The next section provides examples extracted from the many applications that use various high-resolution precipitation products. The final section summarizes the future system for global precipitation processing

Nanobodies Raised against Monomeric α-Synuclein Distinguish between Fibrils at Different Maturation Stages

AbstractNanobodies are single-domain fragments of camelid antibodies that are emerging as versatile tools in biotechnology. We describe here the interactions of a specific nanobody, NbSyn87, with the monomeric and fibrillar forms of α-synuclein (αSyn), a 140-residue protein whose aggregation is associated with Parkinson's disease. We have characterized these interactions using a range of biophysical techniques, including nuclear magnetic resonance and circular dichroism spectroscopy, isothermal titration calorimetry and quartz crystal microbalance measurements. In addition, we have compared the results with those that we have reported previously for a different nanobody, NbSyn2, also raised against monomeric αSyn. This comparison indicates that NbSyn87 and NbSyn2 bind with nanomolar affinity to distinctive epitopes within the C-terminal domain of soluble αSyn, comprising approximately amino acids 118–131 and 137–140, respectively. The calorimetric and quartz crystal microbalance data indicate that the epitopes of both nanobodies are still accessible when αSyn converts into its fibrillar structure. The apparent affinities and other thermodynamic parameters defining the binding between the nanobody and the fibrils, however, vary significantly with the length of time that the process of fibril formation has been allowed to progress and with the conditions under which formation occurs, indicating that the environment of the C-terminal domain of αSyn changes as fibril assembly takes place. These results demonstrate that nanobodies are able to target forms of potentially pathogenic aggregates that differ from each other in relatively minor details of their structure, such as those associated with fibril maturation

A Possible Sterilizing Cure of HIV-1 Infection Without Stem Cell Transplantation

Background: A sterilizing cure of HIV-1 infection has been reported in 2 persons living with HIV-1 who underwent allogeneic hematopoietic stem cell transplantations from donors who were homozygous for the CCR5D32 gene polymorphism. However, this has been considered elusive during natural infection. Objective: To evaluate persistent HIV-1 reservoir cells in an elite controller with undetectable HIV-1 viremia for more than 8 years in the absence of antiretroviral therapy. Design: Detailed investigation of virologic and immunologic characteristics. Setting: Tertiary care centers in Buenos Aires, Argentina, and Boston, Massachusetts. Patient: A patient with HIV-1 infection and durable drug-free suppression of HIV-1 replication. Measurements: Analysis of genome-intact and replication-competent HIV-1 using near-full-length individual proviral sequencing and viral outgrowth assays, respectively; analysis of HIV-1 plasma RNA by ultrasensitive HIV-1 viral load testing. Results: No genome-intact HIV-1 proviruses were detected in analysis of a total of 1.188 billion peripheral blood mononuclear cells and 503 million mononuclear cells from placental tissues. Seven defective proviruses, some of them derived from clonally expanded cells, were detected. A viral outgrowth assay failed to retrieve replication-competent HIV-1 from 150 million resting CD4+ T cells. No HIV-1 RNA was detected in 4.5 mL of plasma. Limitations: Absence of evidence for intact HIV-1 proviruses in large numbers of cells is not evidence of absence of intact HIV-1 proviruses. A sterilizing cure of HIV-1 can never be empirically proved. Conclusion: Genome-intact and replication-competent HIV-1 were not detected in an elite controller despite analysis of massive numbers of cells from blood and tissues, suggesting that this patient may have naturally achieved a sterilizing cure of HIV-1 infection. These observations raise the possibility that a sterilizing cure may be an extremely rare but possible outcome of HIV-1 infection.Fil: Turk, Gabriela Julia Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Seiger, Kyra. Massachusetts Institute of Technology; Estados UnidosFil: Lian, Xiaodong. Massachusetts Institute of Technology; Estados UnidosFil: Sun, Weiwei. Massachusetts Institute of Technology; Estados UnidosFil: Parsons, Elizabeth M.. Massachusetts Institute of Technology; Estados UnidosFil: Gao, Ce. Massachusetts Institute of Technology; Estados UnidosFil: Rassadkina, Yelizaveta. Massachusetts Institute of Technology; Estados UnidosFil: Polo, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Czernikier, Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Ghiglione, Yanina Alexandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Vellicce, Alejandra Fabiana. Universidad de Buenos Aires. Facultad de Medicina. Hospital de Clínicas General San Martín; ArgentinaFil: Varriale, Joseph. No especifíca;Fil: Lai, Jun. No especifíca;Fil: Yuki, Yuko. No especifíca;Fil: Martin, Maureen. No especifíca;Fil: Rhodes, Ajantha. University of Melbourne; AustraliaFil: Lewin, Sharon R.. University of Melbourne; Australia. Monash University; AustraliaFil: Walker, Bruce D.. Massachusetts Institute of Technology; Estados UnidosFil: Carrington, Mary. Massachusetts Institute of Technology; Estados UnidosFil: Siliciano, Robert. No especifíca;Fil: Siliciano, Janet. No especifíca;Fil: Lichterfeld, Mathias. Massachusetts Institute of Technology; Estados UnidosFil: Laufer, Natalia Lorna. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Yu, Xu G.. Massachusetts Institute of Technology; Estados Unido

Internal standard-based analysis of microarray data2—Analysis of functional associations between HVE-genes

In this work we apply the Internal Standard-based analytical approach that we described in an earlier communication and here we demonstrate experimental results on functional associations among the hypervariably-expressed genes (HVE-genes). Our working assumption was that those genetic components, which initiate the disease, involve HVE-genes for which the level of expression is undistinguishable among healthy individuals and individuals with pathology. We show that analysis of the functional associations of the HVE-genes is indeed suitable to revealing disease-specific differences. We show also that another possible exploit of HVE-genes for characterization of pathological alterations is by using multivariate classification methods. This in turn offers important clues on naturally occurring dynamic processes in the organism and is further used for dynamic discrimination of groups of compared samples. We conclude that our approach can uncover principally new collective differences that cannot be discerned by individual gene analysis

Long-Term Blocking of Calcium Channels in mdx Mice Results in Differential Effects on Heart and Skeletal Muscle

The disease mechanisms underlying dystrophin-deficient muscular dystrophy are complex, involving not only muscle membrane fragility, but also dysregulated calcium homeostasis. Specifically, it has been proposed that calcium channels directly initiate a cascade of pathological events by allowing calcium ions to enter the cell. The objective of this study was to investigate the effect of chronically blocking calcium channels with the aminoglycoside antibiotic streptomycin from onset of disease in the mdx mouse model of Duchenne muscular dystrophy (DMD)

A randomised clinical trial of subgrouping and targeted treatment for low back pain compared with best current care. The STarT Back Trial Study Protocol

Back pain is a major health problem and many sufferers develop persistent symptoms. Detecting relevant subgroups of patients with non-specific low back pain has been highlighted as a priority area for research, as this could enable better secondary prevention through the targeting of prognostic indicators for persistent, disabling symptoms. We plan to conduct a randomised controlled trial to establish whether subgrouping using a novel tool, combined with targeted treatment, is better than best current care at reducing long-term disability from low back pain

SmCL3, a Gastrodermal Cysteine Protease of the Human Blood Fluke Schistosoma mansoni

Parasitic infection caused by blood flukes of the genus Schistosoma is a major global health problem. More than 200 million people are infected. Identifying and characterizing the constituent enzymes of the parasite's biochemical pathways should reveal opportunities for developing new therapies (i.e., vaccines, drugs). Schistosomes feed on host blood, and a number of proteolytic enzymes (proteases) contribute to this process. We have identified and characterized a new protease, SmCL3 (for Schistosoma mansoni cathepsin L3), that is found within the gut tissue of the parasite. We have employed various biochemical and molecular biological methods and sequence similarity analyses to characterize SmCL3 and obtain insights into its possible functions in the parasite, as well as its evolutionary position among cathepsin L proteases in general. SmCL3 hydrolyzes major host blood proteins (serum albumin and hemoglobin) and is expressed in parasite life stages infecting the mammalian host. Enzyme substrate specificity detected by positional scanning-synthetic combinatorial library was confirmed by molecular modeling. A sequence analysis placed SmCL3 to the cluster of other cathepsins L in accordance with previous phylogenetic analyses