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Search for microbial signatures within human and microbial calcifications using soft x-ray spectromicroscopy.

BACKGROUND: The origin of advanced arterial and renal calcification remains poorly understood. Self-replicating, calcifying entities have been detected and isolated from calcified human tissues, including blood vessels and kidney stones, and are referred to as nanobacteria. However, the microbiologic nature of putative nanobacteria continues to be debated, in part because of the difficulty in discriminating biomineralized microbes from minerals nucleated on anything else (eg, macromolecules, cell membranes). To address this controversy, the use of techniques capable of characterizing the organic and mineral content of these self-replicated structures at the submicrometer scale would be beneficial. METHODS: Calcifying gram-negative bacteria (Caulobacter crescentus, Ramlibacter tataouinensis) used as references and self-replicating calcified nanoparticles cultured from human samples of calcified aneurysms were examined using a scanning transmission x-ray microscope (STXM) at the Advanced Light Source at Lawrence Berkeley National Laboratory. This microscope uses a monochromated and focused synchrotron x-ray beam (80-2,200 eV) to yield microscopic and spectroscopic information on both organic compounds and minerals at the 25 nm scale. RESULTS: High-spatial and energy resolution near-edge x-ray absorption fine structure (NEXAFS) spectra indicative of elemental speciation acquired at the C K-edge, N K-edge, and Ca L(2,3)-edge on a single-cell scale from calcified C. crescentus and R. tataouinensis displayed unique spectral signatures different from that of nonbiologic hydroxyapatite (Ca(10)(PO(4))(6)(OH)(2)). Further, preliminary NEXAFS measurements of calcium, carbon, and nitrogen functional groups of cultured calcified nanoparticles from humans revealed evidence of organics, likely peptides or proteins, specifically associated with hydroxyapatite minerals. CONCLUSION: Using NEXAFS at the 25 nm spatial scale, it is possible to define a biochemical signature for cultured calcified bacteria, including proteins, polysaccharides, nucleic acids, and hydroxyapatite. These preliminary studies suggest that nanoparticles isolated from human samples share spectroscopic characteristics with calcified proteins.

Aneurysm↗

Genomic signatures of human versus avian influenza A viruses.

Position-specific entropy profiles created from scanning 306 human and 95 avian influenza A viral genomes showed that 228 of 4591 amino acid residues yielded significant differences between these 2 viruses. We subsequently used 15,785 protein sequences from the National Center for Biotechnology Information (NCBI) to assess the robustness of these signatures and obtained 52 "species-associated" positions. Specific mutations on those points may enable an avian influenza virus to become a human virus. Many of these signatures are found in NP, PA, and PB2 genes (viral ribonucleoproteins [RNPs]) and are mostly located in the functional domains related to RNP-RNP interactions that are important for viral replication. Upon inspecting 21 human-isolated avian influenza viral genomes from NCBI, we found 19 that exhibited > or =1 species-associated residue changes; 7 of them contained > or =2 substitutions. Histograms based on pairwise sequence comparison showed that NP disjointed most between human and avian influenza viruses, followed by PA and PB2.

Amino Acid Sequence↗

A proteomic map of thromboinflammatory signatures in antiphospholipid syndrome: results from antiphospholipid syndrome alliance for clinical trials and international networking (APS ACTION) registry.

INTRODUCTION: Antiphospholipid syndrome (APS) is an autoimmune disease with thromboembolic and obstetric morbidity arising via a model of immunothrombosis. Individuals with APS may present with thrombotic (TAPS), obstetric (OAPS), or microvascular (MAPS) disease, while many have circulating antiphospholipid antibodies (aPL) without APS classification (NoAPS). Multiple pathophysiologic mechanisms have been proposed in APS, including activation by aPL of platelets, endothelial and immune cells, as well as complement and coagulation pathways; however, the pathophysiology of APS, particularly transition of clinical APS from aPL remains unclear. METHODS: Seeking to define the inflammatory signature of APS, we carried out an unbiased proteomic screen of persistently aPL-positive patients with different clinical phenotypes from the international APS Alliance for Clinical Trials and International Networking (ACTION) Registry and compared them to 10 healthy controls. 6398 unique proteins were estimated using an DNA aptamer-based assay. Subsequently, we validated our findings in 34 additional patients. RESULTS: Our data show that the mere presence of aPL confers a distinct thromboinflammatory signature characterized by the activation of coagulation, complement, innate and adaptive immune response pathways shared by all APS subtypes. Pathway enrichment analysis revealed increasing enrichment with rising statistical significance of thrombosis, complement, neutrophil and other innate and adaptive immune activation, as well as extracellular matrix (ECM) organization with increasing clinical severity, suggesting a model of progressive thromboinflammation in evolution of APS from NoAPS to TAPS and MAPS. CONCLUSIONS: Our findings provide novel insights into the pathogenesis of APS and identify potential novel targets for diagnostic and therapeutic intervention in APS across its entire spectrum.

Humans↗

Integrated Cytokine and Immune Cell Profiling Reveals a Distinct Immune Signature Associated with High-Altitude Pulmonary Edema.

High-altitude pulmonary edema (HAPE) is a rapidly progressive, life-threatening disorder arising in otherwise healthy individuals upon ascent to high altitude, yet the mechanisms underlying maladaptive vascular leak remain poorly defined. Although elevated pulmonary arterial pressure and capillary stress failure are recognized as central hemodynamic drivers, accumulating evidence indicates that innate immune dysregulation is an equally critical, largely unexplored determinant of HAPE. To systematically delineate the immune and molecular programs that distinguish pathological responses to hypobaric hypoxia from acclimatization, peripheral blood along with clinical details was collected from low-altitude controls (LA-Cntrl, number of participants, (n = 19), healthy high-altitude sojourners (HA-Cntrl, n = 47), and HAPE patients (n = 90). Plasma proteomic markers were quantified using a targeted panel, while monocyte and dendritic cell subsets in peripheral blood mononuclear cells were immunophenotyped by multicolor flow cytometry. HA-Cntrl subjects displayed an anti-inflammatory profile, marked by the suppression of CXC chemokine receptor 3 axis chemokines. HAPE patients, in contrast, exhibited a pro-inflammatory, vascular injury signature, with elevated levels of inflammatory interleukins and myeloid and chemotactic factors. This inflammatory signature was accompanied by the expansion of classical monocytes, implicating a myeloid vascular program associated with HAPE.

Humans↗

Proteomic and Phosphoproteomic Signatures Link Molecular Remodeling to Behavioral Outcomes Following Elderberry and DHA Supplementation in Aging Mice.

Background: Aging is a risk factor for Alzheimer's disease and related dementias, which are associated with synaptic dysfunction and cognitive decline. Elderberry (Sambucus spp.) is rich in anthocyanins with antioxidant and anti-inflammatory properties. Docosahexaenoic acid (DHA), an essential fatty acid, plays a key role in neuronal membrane integrity during brain aging. However, it remains unclear whether elderberry and DHA exert overlapping or distinct effects on brain aging and how these relate to molecular signaling. This study aimed to characterize molecular signatures induced by dietary supplementation and to determine their relationships with behavioral outcomes. Methods: 44-week-old male C57BL/6J mice were randomly assigned to control, elderberry, DHA, or combined diets for 12 weeks. Behavioral testing assessed anxiety-like behavior, spatial learning and memory. Brain tissues underwent proteomic and phosphoproteomic profiling and fatty-acid analysis. Data were analyzed using Ingenuity Pathway Analysis to identify enriched pathways, upstream regulators, and functional associations. Results: Elderberry as well as DHA supplementation induced targeted remodeling of the proteome and phosphoproteome, with pathway enrichment involving synaptogenesis, glutamatergic signaling, and long-term potentiation. Upstream-regulator analysis predicted elderberry-associated CDK5 signaling, accompanied by reduced MAPT/Tau phosphorylation at selected sites, whereas DHA supplementation was associated with CAMK-related signaling. DHA supplementation altered fatty-acid composition, increasing the n-3/n-6 ratio. Elderberry reduced anxiety-like behavior and improved target-directed search during the Barnes maze probe test. Molecular signatures were examined in relation to the measured behavioral outcomes. Conclusions: Elderberry and DHA are associated with distinct molecular networks related to synaptic function and behavioral outcomes in the aging male mouse brain. These findings support further investigation of elderberry and DHA as dietary interventions targeting molecular and behavioral features of brain aging.

Animals↗

Transcript signatures in experimental asthma: identification of STAT6-dependent and -independent pathways.

The analysis of polygenic diseases such as asthma poses a challenging problem. In an effort to provide unbiased insight into disease pathogenesis, we took an empirical approach involving transcript expression profiling of lung tissue from mice with experimental asthma. Asthmatic responses were found to involve sequential induction of 4.7% of the tested genome; notably, there was ectopic expression of a series of genes not previously implicated in allergic or pulmonary responses. Genes were widely distributed throughout all chromosomes, but preferentially included genes involved in immunity, development, and homeostasis. When asthma was induced by two independent experimental regimens, unique gene transcript profiles were found depending upon the mode of disease induction. However, the majority of genes were common to both models representing an asthma signature genome. Analysis of STAT6-deficient mice revealed that an unexpectedly large segment of the asthma genes were STAT6 independent; this correlated with sustained inflammatory events in these mice. Notably, induction of asthma in STAT6-deficient mice resulted in gene induction not seen in wild-type mice. These results raise concern that therapeutic blockade of STAT6 in the asthmatic setting may reprogram the genetic signature, resulting in alternative lung pathology, which we indeed observed in STAT6-deficient mice. These results provide unprecedented insight into the complex steps involved in the pathogenesis of allergic airway responses; as such, these results have significant therapeutic and clinical implications.

Administration, Intranasal↗

Anti-HIV state but not apoptosis depends on IFN signature in CD4+ T cells.

To gain insights into the molecular mechanisms underlying early host responses to HIV in the CD4(+) T cell target population, we examined gene expression in CD4(+) T cells isolated 24 h after ex vivo HIV infection of lymphocyte aggregate cultures derived from human tonsils. Gene profiling showed a distinct up-regulation of genes related to immune response and response to virus, notably of IFN-stimulated genes (ISGs), irrespective of the coreceptor tropism of the virus. This mostly IFN-alpha-dependent gene signature suggested the involvement of plasmacytoid dendritic cells, a principal component of the antiviral immune response. Indeed, depletion of plasmacytoid dendritic cells before HIV inoculation abrogated transcriptional up-regulation of several ISGs and resulted in increased levels of HIV replication. Treatment with a blocking anti-IFN-alphaR Ab yielded increased HIV replication; conversely, HIV replication was decreased in pDC-depleted cultures treated with IFN-alpha. Among up-regulated ISGs was also TRAIL, indicating a potential role of the IFN signature in apoptosis. However, a blocking anti-TRAIL Ab did not abrogate apoptosis of CD4(+) T cells in CXCR4-tropic HIV-infected cultures, suggesting the involvement of pathways other than TRAIL mediated. We conclude that acute HIV infection of lymphoid tissue results in up-regulation of ISGs in CD4(+) T cells, which induces an anti-HIV state but not apoptosis.

Antibodies, Blocking↗

Electronic signature of computerized patient records.

The article describes the computerized system developed and used at the Ohio State University Medical Center in Columbus, Ohio to affix a physician's signature electronically. The system electronically sends dictated reports (e.g., discharge summaries, history and physical examinations, and operative reports) and attestation statements to the physician for review. The physician selects the report individually and may approve or enter comments. Comments are forwarded electronically to medical information management. The article describes the fiscal intermediary's requirements and approval process and the impact on delinquent records and accounts awaiting attestation signature before billing.

Authorship↗

HIPAA, security, and electronic signature: a closer look.

A recent notice of proposed rule making for an electronic signature standard, if adopted, will affect the way health information is managed for years to come. Here's what you need to know about electronic signature and what the proposed standard may mean for HIM.

Authorship↗

Recovery of human skeletal elements from a recent UK murder inquiry: preservational signatures.

Factors that control bone preservation are not fully understood but generally include those that reflect "natural" taphonomic or diagenetic processes and also those reflecting anthropogenic activity. The aim of this paper is to examine whether the survival of skeletal elements from a recent UK serial murder investigation (n = 12) and three archaeological cemetery sites from England (n = 112, 95, 182; Roman to early-medieval), share a similar recovery signature. Examination of this data demonstrates that even when clear evidence of traumatic and perimortem dismemberment exists within an assemblage, the distribution of missing elements can be almost identical to archaeological material buried in normal attrition cemeteries. Given that these preservational signatures are so similar, it is concluded that careful observation of bone surfaces is necessary to confidently interpret bone loss, particularly where dismemberment and/or element excision is suggested by the non-anatomical position of the skeleton within the grave. Where postmortem excision of bone is suspected, careful examination of contiguous bone surfaces, both macroscopic and microscopic, is suggested to detect fine cutmark lesions indicative of anthropogenic excision. Without this evidence other preservational factors must be considered both taphonomic and diagenetic.

Adolescent↗

The Doctrine of Signatures: a historical, philosophical, scientific view (II).

The first part of this article discussed the history of the Doctrine of Signatures. Bach developed a series of Flower Remedies for emotional states. On the basis of the Doctrine of Signatures, these can liberate the vital force from an alien disease process or 'archeus'. An understanding of the role of the active principle of a homeopathic medicine, in the physiology of its source, may aid understanding of a curative action.

Homeopathy↗

Insonation method and diagnostic flow signatures for transcranial power motion (M-mode) Doppler.

BACKGROUND AND PURPOSE: Power motion mode Doppler (PMD) simultaneously displays flow signal intensity and direction over several centimeters of intracranial space. Insonation protocol for PMD and spectral transcranial Doppler (TCD) with typical PMD flow signatures is described in serial patients with acute stroke symptoms examined via conventional windows with a PMD/TCD unit. RESULTS: Thirty-five patients were studied within 12 hours after stroke onset (age 64 +/- 15 years; 8 received intravenous and 3 intra-arterial thrombolysis). One patient had no temporal window, and 3 patients had suboptimal windows. In 90% of patients, PMD showed more than 1 ipsilateral temporal windows. In 63% of patients (n = 22), PMD simultaneously displayed the entire M1 (65-45 mm) and proximal M2 (45-30 mm) flows, leading to spectral TCD examination of the proximal M2 middle cerebral artery (MCA) in 28 of 35 patients (80%). All patients had sufficient foraminal (depth display = 60-110 mm) and orbital (depth display = 30-80 mm) windows. PMD displayed the entire basilar artery stem (75-100+ mm) in 69% (n = 24) of patients, and the distal basilar flow was detected in all patients by both PMD and TCD. TCD results were normal (12), proximal intracranial stenosis (5), large vessel occlusion (17), and cerebral circulatory arrest (1). Compared to spectral TCD, PMD signatures of similar diagnostic significance were low resistance (vessel identification and recanalization), high resistance (ophthalmic artery identification and distal obstruction), collateral (communicating arteries and leptomeningeal flow), reverberating (circulatory arrest), and branch embolization. CONCLUSIONS: PMD is a window-finding tool and a guide for spectral TCD gate placement. PMD facilitates flow detection in the M2 branches and the distal basilar artery. PMD can demonstrate recanalization of the entire MCA main stem and proximal branches, increase the yield of embolus detection and procedure monitoring, and facilitate abnormal flow pattern recognition.

Aged↗

Mutational signature of the proximate bladder carcinogen N-hydroxy-4-acetylaminobiphenyl: inconsistency with the p53 mutational spectrum in bladder cancer.

We studied the mutagenicity of the proximate bladder carcinogen, N-hydroxy-4-acetylaminobiphenyl (N-OH-AABP) in embryonic fibroblasts of the Big Blue mouse. Treatment of these cells with increasing concentrations of N-OH-AABP for 24 h resulted in a dose-dependent increase in mutation frequency of the cII transgene up to 12.8-fold over the background. Single base substitutions comprised 86% of the N-OH-AABP-induced mutations and 74% of the spontaneous cII mutations (sequenced number of mutant plaques, 141 and 145, respectively). Of these, 63 and 36%, respectively, occurred at guanine residues along the cII gene. Whereas G to T transversions predominated in the induced cII mutations (47%), insertion was the most spontaneously derived cII mutation (19%). Mapping of N-OH-AABP-induced DNA adducts along the cII gene by terminal transferase-dependent PCR showed the formation of DNA adducts at specific nucleotide positions. Five preferential DNA adduction sites were established, of which four were major mutation sites for N-OH-AABP, especially for G to T transversions. This unique mutational signature of N-OH-AABP in the cII gene was, however, in sharp contrast with the mutational spectrum of the p53 gene in human bladder cancer. G to A transitions are the dominant type of p53 mutations (53%), being also prevalent in almost all of its five mutational hotspots (codons 175, 248, 273, 280, and 285). In addition, the majority of mutations in three of these hotspots (codons 175, 248, and 273) are at a methylated CpG site, whereas in the cII gene neither the preferential N-OH-AABP DNA adduction sites nor the induced mutational hotspots are biased toward methylated CpG dinucleotides. We conclude that N-OH-AABP leaves a characteristic mutational signature in the cII transgene, which is consistent with its preferential DNA adduction profile. However, the pattern of mutation induced by N-OH-AABP in the cII gene is largely at odds with the mutational spectrum of the p53 gene in human bladder cancer.

Aminobiphenyl Compounds↗

[Some evidence for the use of doctrine of signatures in the land of Israel and its environs during the Middle Ages].

The Doctrine of Signatures, was developed in Europe in the 16th and 17th centuries, though, its traces are spread until the present day in different traditional medicine cultures. This study traces the use of the Doctrine of Signatures in the medical and pharmacological literature of the Land of Israel and it's environs during the Middle Ages. The historical sources support the claim that although this theory did not originate in the region, it was certainly practiced there. These sources have revealed 23 substances with medicinal uses based on the Doctrine, bearing witness to the extent of its influence at the time: PLANTS: Common Agrimony, Common Balm, Common Snapdragon, Coral Peony, Corn Gromwell, Lebanon Barberry, Mullein, Orchid, Panther Strangler, Red Horned Poppy, Rhubarb, Rose of Jericho, Southern Maidenhair Fern, Spiny Broom, Sumach, Walnut, Wild Dog Rose. ANIMALS: Firefly, Red Coral, White Cuttle Fish. MINERALS: Red Chalk (Haematite), Sea Urchin, White Clay. The main categories of the Doctrine uncovered were: similarity between the substance used and the human organ; resemblance in shape or behaviour to a specific animal; correlation between the colour of a substance and the colour of the symptoms; similarities between the substance and the patient's symptoms and the use of a substance that might produce symptoms of a particular disease in a healthy person to remedy those same symptoms in one who is sick.

Animals↗

Electronic data management for the Hemochron Jr. Signature coagulation analyzer.

Point-of-care testing (POC, POCT) laboratory devices are being introduced into operating suites and critical care units in ever increasing numbers. The small, portable devices have gained in popularity because of their ease of use and the rapid availability of test results. POCT is an integral part of extracorporeal technology (ECT). A challenge associated with the growth of POC technology is related to management of the data generated by these devices. In the field of ECT, storing, retrieving, analyzing, viewing and charting quality control (QC) and patient test data generated with POC coagulation instruments is essential. We evaluated a premarket version of data management software developed for the Hemochron Jr. Signature coagulation analyzer, a PC-based software capable of fulfilling our objective. A database comprised of greater than 50 plasma and electronic QC results and greater than 140 patient sample results for ACT, PT, and aPTT tests was transferred from a Hemochron Jr. Signature device to two different PCs, each equipped with Hemochron ReportMaker software supplied by the manufacturer. Data files were transferred directly from the coagulation test unit to the PCs via an RS-232 cable. A variety of charts, reports, and file listings were created from the datasets using the software menus. Transfer of the complete database required less than 5 min. The relative speed and simplicity of the data interface promotes frequent charting of QC data, permitting real-time monitoring and early identification of data trends or values requiring intervention. If a subset of QC data is found to be incomplete, altered, or unacceptable, all patient samples tested during that period can be promptly identified. The software also includes data query tools useful for sorting and selecting specific subsets of patient and QC data. Electronic data management can facilitate compliance with quality control requirements and assist clinicians and laboratory personnel in the collection, storage, and review of quality control and patient test data. In addition, the patient and QC data are readily accessible when necessary for use in risk management assessment, accreditation, or litigation proceedings.

Blood Coagulation Tests↗

New FDA guidance on electronic records and signatures.

The United States Food and Drug Administration (FDA) is in the process of developing and issuing a series of guidance documents on electronic records and signatures requirements. This article discusses two recently published FDA guidance documents: validation requirements for electronic records and signatures and a glossary of terms. It also highlights the need for industry to provide comments on these documents to FDA.

Authorship↗

Fatty acid synthase expression defines distinct molecular signatures in prostate cancer.

The androgen-regulated enzyme fatty acid synthase (FAS), required for de novo lipogenesis, is overexpressed in several cancers including prostate carcinoma and has been associated with aggressive disease. FAS expression was assessed in 81 prostate carcinomas, both by immunohistochemistry in tissue microarrays and by Affymetrix Hu95Av2 oligonucleotide arrays. Both FAS mRNA and protein were significantly overexpressed in prostate carcinomas compared with the corresponding normal tissue. FAS mRNA and protein expression increased substantially from normal to prostatic intraepithelial neoplasia, to low grade, to high grade, and to androgen-independent bone metastases. A significant correlation between FAS mRNA and protein expression was found in two thirds of the cases. In 17% of the cases, FAS protein levels were high despite low mRNA levels, and these tumors exhibited a distinct molecular signature when compared with tumors that did not express FAS protein. Whereas the latter group of tumors expressed some proapoptotic genes, tumors with high FAS levels overexpressed, among other genes, its transcriptional regulator, steroid regulator binding protein, and apolipoprotein E. These data demonstrate (1) the consistent overexpression of FAS in prostate carcinoma compared with the adjacent normal tissue, (2) a strong association between FAS and prostate tumor initiation and progression, (3) the highest FAS expression occurring in androgen-independent bone metastases, (4) the transcriptional and posttranscriptional regulation of FAS in the majority and in a subset of prostate cancers, respectively, and (5) most importantly, the identification by FAS expression of prostate tumors with unique molecular signatures and potentially diverse biologic behavior.

Androgens↗

Electronic signatures for long-lasting storage purposes in electronic archives.

Communication and co-operation in healthcare and welfare requires certain Trusted Third Party (TTP) services describing both status and relation of communicating principals as well as their corresponding keys and attributes. Additional TTP services are needed to provide trustworthy information about dynamic issues of communication and co-operation such as time and location of processes, workflow relations, and system behaviour. Electronic signatures based on asymmetric cryptography are important means for securing the integrity of a message or file as well as for accountability purposes including non-repudication of both origin and receipt. These electronic signatures along with certified time stamps are especially important for electronic health records (EHR), electronic archives in general, and other typical purposes of a long-lasting storage. Apart from technical storage problems (e.g. lifetime of the storage devices), this paper needs to look for mechanisms of e.g. re-signing of messages, archive details, or whole archives.

Access to Information↗