No description available.

Protein phosphorylation is a reversible post-translation modification that is essential in cell signaling. It is estimated that a third of all cellular proteins are phosphorylated (reviewed in Ficarro et al., 2002), with more than 98% of those phosphorylation events occurring on serine and threonine residues (Olsen et al., 2006). Kinases are the necessary enzymes for phosphorylation and protein phosphatases dynamically reverse this action. While the mechanisms of substrate recognition for kinases have been well-characterized to date, the same is not true for phosphatases that play an equally important role in opposing kinase function and determining global phosphorylation levels in cells. This dichotomy has also translated into the clinic, where there has been a persistently narrow research focus on the development of small-molecule kinase inhibitors for use as chemotherapeutic agents, without an equal effort being placed into the generation of the analogous phosphatase activators (reviewed in Westermarck, 2018). Members of the phosphoprotein phosphatase (PPP) family of serine/threonine phosphatases are responsible for the majority of dephosphorylation in eukaryotic cells, with protein phosphatase 1 (PP1) and protein phosphatase 2A (PP2A) accounting for more than 90% of the total phosphatase activity (Moorhead et al., 2007; Virshup and Shenolikar, 2009). Structurally, PP2A is a trimeric holoenzyme consisting of a scaffold (A) subunit, a regulatory (B) subunit, and a catalytic (C) subunit. B55α is a ubiquitous regulatory subunit that is reported to target many substrates with critical functions in processes including cell division. A long-standing question that has persisted in the field of cellular signaling is as to how the most abundant serine/threonine PP2A holoenzyme, PP2A/B55α, specifically recognizes substrates and presents them to the enzyme active site for subsequent dephosphorylation. Such critical data have only recently become well understood for the B56 family of ‘B’ regulatory subunits, where an LxxIxE short linear motif (or SLiM) has been identified in a subset of protein targets and shown via crystal structure analysis to dock into a 100% conserved binding pocket on the B56 surface (Hertz et al., 2016; Wang et al., 2016a; Wang et al., 2016b; Wu et al., 2017). Here, we show how B55α recruits p107, a pRB-related tumor suppressor and B55α substrate. Using molecular and cellular approaches, we identified a conserved region 1 (R1, residues 615-626) encompassing the strongest p107 binding site. This enabled us to identify an “HxRVxxV619-625” SLiM in p107 as necessary for B55α binding and dephosphorylation of the proximal pSer-615 in vitro and in cells. Numerous additional PP2A/B55α substrates, including TAU, contain a related SLiM C-terminal from a proximal phosphosite, allowing us to propose a consensus SLiM sequence, “p[ST]-P-x(5-10)-[RK]-V-x-x-[VI]-R”. In support of this, mutation of conserved SLiM residues in TAU dramatically inhibits dephosphorylation by PP2A/B55α, validating its generality. Moreover, a data-guided computational model details the interaction of residues from the conserved p107 SLiM, the B55α groove, and phosphosite presentation to the PP2A/C active site. Altogether, these data provide key insights into PP2A/B55α mechanisms of substrate recruitment and active site engagement, and also facilitate identification and validation of new substrates, a key step towards understanding the role of PP2A/B55α in many key cellular processes. As a parallel continuation of our efforts to identify novel B55α substrates/regulators, we generated mutant B55α constructs that occlude PP2A/A-C dimer engagement but retain substrate binding to the β-propeller structure (allowing us to interrogate direct interactors). Our preliminary AP-MS data led to the identification of several proteins that bound better to our “monomeric B55α” mutant compared to wild-type B55α in the context of the PP2A/B55α heterotrimer, including the centrosomal proteins HAUS6 and CEP170 (two substrates previously validated in a phosphoproteomic screen by our lab), suggesting that these mutants trap substrates as they cannot be dephosphorylated by PP2A/C. These analyses also identified an enrichment of T-complex protein 1 subunits in the “monomeric B55α” mutant elutions, further supporting the notion that these mutants may function as dominant negatives. Several additional proteins of interest were identified in the two independent rounds of mass spectrometry, including subunits of the DNA-directed RNA polymerases I, II, and IV, as well as the double-strand break repair protein MRE11, which can be followed up as potential novel B55α substrates. These studies can contribute to significant advances in our understanding of the network of proteins that B55α interacts with, and thus the signaling pathways that can be modulated by PP2A/B55α complexes in cells. Moreover, these advances can also provide translational benefits as has been demonstrated through the study of PP2A activators termed SMAPs, which demonstrate selective stabilization of PP2A/B56α complexes in cells that result in selective dephosphorylation of substrates including the oncogenic target c-MYC. / Biomedical Sciences

No description available.

Eighteen well trained males (1RM Squat: 150.57 ± 26.79 kg) were assigned to two groups: full ROM training (control) and full ROM with partial ROM training (CP) for the seven-week training intervention. There was a significant time effect (p

Documentation exists that mothers can pass the cariogenic bacteria Streptococcus mutans to their infants. The newest technology to identify Streptococcus mutans is a rapid detection saliva test. Two hundred patients above the age of 18 were targeted using random selection in a Louisville, Kentucky dental office. Patients signed an informed consent form and were given a qualifying questionnaire. Patients received 2 bitewing x-rays and a charted DMFT index and were administered the saliva test. While the null hypothesis was rejected using the chi square test, the results were inconclusive due to expected values. However, other chi square results revealed that the test worked or had the potential to work. Furthermore, it was concluded that the test had high specificity. Further research is warranted; however, the saliva test in combination with the DMFT and x-rays are instrumental tools for the dental professional in educating patients and prevention.

Abstract available in the Clinical Pharmacology in Drug Development.

No description available.

THE PROPER APPLICATION OF THE PRINCIPLE OF SPECIFICITY IS ESSENTIAL TO ANY STRENGTH AND CONDITIONING PROGRAM. HOWEVER, THE TRANSFER OF RESISTANCE TRAINING TO SPORT IS HIGHLY COMPLEX, DIFFICULT TO PREDICT, AND CHALLENGING TO ASSESS. THIS BRIEF REVIEW EXAMINES THE PRINCIPLE OF DYNAMIC CORRESPONDENCE AS AN AID TOWARD BETTER UNDERSTANDING AND PREDICTING AN EXERCISE OR TRAINING METHOD'S POTENTIAL TRANSFER TO SPORT. PRACTICAL TRAINING RECOMMENDATIONS ARE GIVEN BASED ON THE RESEARCH REVIEWED.

Lyme borreliosis, more commonly referred to as Lyme disease, is the fastest growing zoonotic disease in North America with approximately 30,000 confirmed cases and 300,000 estimated infections per year. In nature, the causative agent of Lyme disease, the bacterium Borrelia burgdorferi, cycles between Ixodes sp. ticks and small mammals. Humans become infected with Lyme disease after being bitten by an infected tick. The primary indicator of a Borrelia burgdorferi infection is a bull’s eye rash typically followed by flu-like symptoms with treatment consisting of a 2-4 week course of antibiotics. If not treated, later stages of the disease can result in arthritis, cardiovascular and neurological symptoms. Diagnosis of Lyme disease is challenging and currently requires a complex laboratory diagnostic using indirect detection of host-generated antibodies by a two-tiered approach consisting of an enzyme linked immunosorbent assay (ELISA) followed by IgM and IgG immunoblots. Although two-tier testing has provided an adequate approach for Lyme disease diagnosis, it has weaknesses including subjective analysis, complex protocols and lack of reagent standardization. Immuno-PCR (iPCR) is a method that combines ELISA-based detection specificity with the sensitivity of PCR signal amplification and has demonstrated increased sensitivity for many applications such as detection of disease biomarkers but has yet to be applied for diagnosis of Lyme disease. Herein, using iPCR and recombinant B. burgdorferi antigens, an assay for both the direct and the indirect detection of Lyme disease was developed iv and demonstrated improved sensitivity for detection of B. burgdorferi antibodies using a murine model. Moreover, we present evidence using human Lyme disease patient serum samples that iPCR using both multiple antigens and a unique single hybrid antigen is capable of achieving increased sensitivity and specificity compared to existing methodology. These data represent the first demonstration of iPCR for Lyme disease diagnosis and support the replacement of two-tier testing with a more simplified and objective approach.

At Santa Margarita Ranch, California, oak mistletoe (Phoradendron villosum) parasitizes valley oak and blue oak but cannot be found growing on coast live oak despite its abundance and ability to parasitize coast live oak in other areas. It seems as though this species of mistletoe is specializing on certain host oak trees, but the mechanisms of this specialization are unknown. In order to investigate this pattern, we utilized a type of machine learning in GIS called supervised classification as well as a reciprocal transplant study in the field. The three species of oak trees were classified with 87% accuracy using drone imagery and 95% accuracy using open source NAIP imagery. This classification technique could be applied to the whole state of California as long as ground truth points for each species were collected. This could be extremely useful for large scale forest management projects and ecological questions. Unfortunately, the classifier was unsuccessful at distinguishing mistletoe from host and so the number of mistletoe on each host could not be quantified using this technique. The reciprocal transplant study involved collecting mistletoe fruit from individuals growing on each of the three hosts and experimentally applying them back onto all three hosts. This allowed us to test whether there are host races of mistletoe that specialize at growing on certain hosts. We found that seeds from each host origin germinated equally well regardless of where they were dispersed, and seeds survived best on coast live oak, regardless of where they originated from. Based on these results, there must be some mechanism, other than host races, that explains the lack of mistletoe on coast live oaks at Santa Margarita Ranch. Future projects should investigate whether evidence for host races can be found at a later stage of seedling development and the roll of bird dispersers in creating the pattern.