Prostate cancer is a clinically heterogeneous disease, ranging from indolent asymptomatic disease to very aggressive metastatic and life threatening forms of the disease. Distant metastasis ...represents the major lethal cause of prostate cancer. The most critical clinical challenge in the management of the patients is identifying those individuals at risk of developing metastatic disease. To understand the molecular mechanisms of prostate cancer metastasis and identify markers with metastatic potential, we have analyzed protein expression in two syngeneic prostate cancer cells lines PC3-N2 and PC3-ML2 using isobaric tags for relative and absolute quantitation labeling and multi-dimensional protein identification technology liquid chromatography matrix assisted laser desorption ionization tandem mass spectrometry. PC3-N2 is lowly metastatic while PC3-ML2 highly metastatic. A total of 1,756 proteins were identified in the analyses with 130 proteins showing different expression levels (p<0.01) in the two cell lines. Out of these, 68 proteins were found to be significantly up-regulated while 62 are significantly down-regulated in PC3-ML2 cells compared with PC3-N2 cells. The upregulation of plectin and vimentin which were the most significantly differentially expressed were validated by Western blot and their functional relevance with respect to invasion and migration was determined by siRNA gene silencing. To our knowledge, this study is the first to demonstrate that up-regulation of vimentin and plectin expression positively correlates with the invasion and metastasis of androgen-independent PCA.
Wealth building programs remain underutilized, and Medical Financial Partnerships serve as a potential solution. We aimed to assess the reach and adoption of an underutilized asset building program, ...Family Self Sufficiency, with a national uptake of 3%, when integrated into a healthcare system.
First, a hospital-affiliated "known provider" introduced Family Self Sufficiency to clinic patients. Second, hospital staff unknown to families conducted outreach to clinic patients. For both pilots, we tracked eligibility, interest, and enrollment rates. We evaluated the pilots using the Reach, Effectiveness, Adoption, Implementation, and Maintenance framework in addition to reviewing the qualitative feedback from the staff who introduced the program.
The reach of each pilot varied: the first pilot (n = 17) had an enrollment rate of 18%, whereas the second pilot (n = 69) had an enrollment rate of 1%. Adoption factors included prior relationship with the family and barriers to understanding the program families. However, adoption was limited by bandwidth of family to complete paperwork, staff to do outreach, and timing of the outreach to maximize benefit.
Increasing uptake of underutilized asset building programs could be part of the solution to building wealth for families with low incomes. Healthcare partnerships may be an approach to increase reach and adoption by eligible populations. Areas to consider for successful future implementation include: (1) timeline of outreach, (2) families' relationship with individuals performing outreach, and (3) current bandwidth of the family. Systematic implementation trials are needed to study these outcomes in more detail.
Objectives
Widespread global transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus causing coronavirus disease 2019 (COVID-19), continues. Many questions remain ...about asymptomatic or atypical infections and transmission dynamics. We used comprehensive contact tracing of the first 2 confirmed patients in Illinois with COVID-19 and serologic SARS-CoV-2 antibody testing to determine whether contacts had evidence of undetected COVID-19.
Methods
Contacts were eligible for serologic follow-up if previously tested for COVID-19 during an initial investigation or had greater-risk exposures. Contacts completed a standardized questionnaire during the initial investigation. We classified exposure risk as high, medium, or low based on interactions with 2 index patients and use of personal protective equipment (PPE). Serologic testing used a SARS-CoV-2 spike enzyme-linked immunosorbent assay on serum specimens collected from participants approximately 6 weeks after initial exposure to either index patient. The 2 index patients provided serum specimens throughout their illness. We collected data on demographic, exposure, and epidemiologic characteristics.
Results
Of 347 contacts, 110 were eligible for serologic follow-up; 59 (17% of all contacts) enrolled. Of these, 53 (90%) were health care personnel and 6 (10%) were community contacts. Seventeen (29%) reported high-risk exposures, 15 (25%) medium-risk, and 27 (46%) low-risk. No participant had evidence of SARS-CoV-2 antibodies. The 2 index patients had antibodies detected at dilutions >1:6400 within 4 weeks after symptom onset.
Conclusions
In serologic follow-up of the first 2 known patients in Illinois with COVID-19, we found no secondary transmission among tested contacts. Lack of seroconversion among these contacts adds to our understanding of conditions (ie, use of PPE) under which SARS-CoV-2 infections might not result in transmission and demonstrates that SARS-CoV-2 antibody testing is a useful tool to verify epidemiologic findings.
We have designed and built a magnetic tweezers device that enables the application of calibrated stresses to soft materials while simultaneously measuring their microscale deformation using confocal ...microscopy. Unlike previous magnetic tweezers designs, our device is entirely portable, allowing easy use on microscopes in core imaging facilities or in collaborators' laboratories. The imaging capabilities of the microscope are unimpaired, enabling the 3-D structures of fluorescently labeled materials to be precisely determined under applied load. With this device, we can apply a large range of forces (∼1-1200 pN) over micron-scale contact areas to beads that are either embedded within 3-D matrices or attached to the surface of thin slab gels. To demonstrate the usefulness of this instrument, we have studied two important and biologically relevant materials: polyacrylamide-based hydrogel films typical of those used in cell traction force microscopy, and reconstituted networks of microtubules, essential cytoskeletal filaments.
Using prostatic fluids rich in glycoproteins like prostate-specific antigen and prostatic acid phosphatase (PAP), the goal of this study was to identify the structural types and relative abundance of ...glycans associated with prostate cancer status for subsequent use in emerging MS-based glycopeptide analysis platforms.
A series of pooled samples of expressed prostatic secretions (EPS) and exosomes reflecting different stages of prostate cancer disease were used for N-linked glycan profiling by three complementary methods, MALDI-TOF profiling, normal-phase HPLC separation, and triple quadropole MS analysis of PAP glycopeptides.
Glycan profiling of N-linked glycans from different EPS fluids indicated a global decrease in larger branched tri- and tetra-antennary glycans. Differential exoglycosidase treatments indicated a substantial increase in bisecting N-acetylglucosamines correlated with disease severity. A triple quadrupole MS analysis of the N-linked glycopeptides sites from PAP in aggressive prostate cancer pools was done to cross-reference with the glycan profiling data.
Changes in glycosylation as detected in EPS fluids reflect the clinical status of prostate cancer. Defining these molecular signatures at the glycopeptide level in individual samples could improve current approaches of diagnosis and prognosis.
Spatial representations of threatening processes - "threat maps" - can identify where biodiversity is at risk, and are often used to identify priority locations for conservation. In doing so, ...decision makers are prone to making errors, either by assuming that the level of threat dictates spatial priorities for action or by relying primarily on the location of mapped threats to choose possible actions. We show that threat mapping can be a useful tool when incorporated within a transparent and repeatable structured decision-making (SDM) process. SDM ensures transparent and defendable conservation decisions by linking objectives to biodiversity outcomes, and by considering constraints, consequences of actions, and uncertainty. If used to make conservation decisions, threat maps are best developed with an understanding of how species respond to actions that mitigate threats. This approach will ensure that conservation actions are prioritized where they are most cost-effective or have the greatest impact, rather than where threat levels are highest.
A cardinal symptom of major depressive disorder (MDD) is the disruption of circadian patterns. However, to date, there is no direct evidence of circadian clock dysregulation in the brains of patients ...who have MDD. Circadian rhythmicity of gene expression has been observed in animals and peripheral human tissues, but its presence and variability in the human brain were difficult to characterize. Here, we applied time-of-death analysis to gene expression data from high-quality postmortem brains, examining 24-h cyclic patterns in six cortical and limbic regions of 55 subjects with no history of psychiatric or neurological illnesses (“controls”) and 34 patients with MDD. Our dataset covered ∼12,000 transcripts in the dorsolateral prefrontal cortex, anterior cingulate cortex, hippocampus, amygdala, nucleus accumbens, and cerebellum. Several hundred transcripts in each region showed 24-h cyclic patterns in controls, and >100 transcripts exhibited consistent rhythmicity and phase synchrony across regions. Among the top-ranked rhythmic genes were the canonical clock genes BMAL1(ARNTL), PER1-2-3, NR1D1(REV-ERBa), DBP, BHLHE40 (DEC1) , and BHLHE41(DEC2) . The phasing of known circadian genes was consistent with data derived from other diurnal mammals. Cyclic patterns were much weaker in the brains of patients with MDD due to shifted peak timing and potentially disrupted phase relationships between individual circadian genes. This transcriptome-wide analysis of the human brain demonstrates a rhythmic rise and fall of gene expression in regions outside of the suprachiasmatic nucleus in control subjects. The description of its breakdown in MDD suggests potentially important molecular targets for treatment of mood disorders.
While ketones are among the most versatile functional groups, their synthesis remains reliant upon reactive and low‐abundance starting materials. In contrast, amide formation is the most‐used ...bond‐construction method in medicinal chemistry because the chemistry is reliable and draws upon large and diverse substrate pools. A new method for the synthesis of ketones is presented here that draws from the same substrates used for amide bond synthesis: amines and carboxylic acids. A nickel terpyridine catalyst couples N‐alkyl pyridinium salts with in situ formed carboxylic acid fluorides or 2‐pyridyl esters under reducing conditions (Mn metal). The reaction has a broad scope, as demonstrated by the synthesis of 35 different ketones bearing a wide variety of functional groups with an average yield of 60±16 %. This approach is capable of coupling diverse substrates, including pharmaceutical intermediates, to rapidly form complex ketones.
Amine+carboxylic acid=ketone: Diverse amines and carboxylic acids can be coupled to rapidly form complex ketones. The keys to this discovery are: 1) the use of alkyl pyridinium salts, which are formed in one step from amines, as alkyl radical donors; 2) the use of in situ generated acyl fluorides or 2‐pyridyl esters as acyl donors; and 3) a nickel catalyst with a terpyridine ligand to unite the two intermediates.
A nickel-catalyzed reductive cross-coupling of alkylpyridinium salts and aryl bromides has been developed using Mn as the reductant. Both primary and secondary alkylpyridinium salts can be used, and ...high functional group and heterocycle tolerance is observed, including for protic groups. Mechanistic studies indicate the formation of an alkyl radical, and controlling its fate was key to the success of this reaction.
Highlights • We have used the prosomeric model to create a modern ontology of mammalian brain structures. • This ontology is based chiefly on gene expression during development. • This ontology will ...be useful to the field of neuroinformatics.
PDF
