Regulation of Human Epidermal Keratinocyte Differentiation by the Vitamin D Receptor and its Coactivators DRIP205, SRC2, and SRC3

It has long been known that the active metabolite of vitamin D, 1,25 dihydroxyvitamin D3, stimulates differentiation and inhibits proliferation in epidermal keratinocytes through interaction with the vitamin D receptor (VDR). VDR functions through the coordinate binding of vitamin D response elements in the DNA and specific coactivator proteins which help to initiate transcription. It was recently observed that VDR binds to two major coactivator complexes, DRIP (VDR-interacting protein) and SRC (steroid receptor coactivator), during keratinocyte differentiation. To determine the role of VDR and its coactivators in mediating keratinocyte differentiation, we developed an adenoviral system to knock down, or in the case of VDR, overexpress these genes. In order to study all stages of keratinocyte development, we employed an advanced differentiated normal human keratinocyte culture system that produces a multilayer phenotype similar to that of normal skin. These studies have shown that VDR, DRIP, and SRC are all required for promotion of both early and late keratinocyte differentiation. Additionally, each individual differentiation marker that was assayed has a different specificity for the coactivators that regulate its expression

Bioluminescence, photophysical, computational and molecular docking studies of fully conformationally restricted enamine infraluciferin

A new rationally designed fully rotationally restricted luciferin has been synthesised. This synthetic luciferin, based upon the structure of infraluciferin, has two intramolecular H-bonds to reduce degrees of freedom, an amine group to enhance ICT process, and an alkenyl group to increase π-conjugation. In the spectroscopic measurements and computational calculations, enamine luciferin showed more red-shifted absorption and fluorescence emission than LH2 and iLH2. With PpyWT luciferase enamine luciferin gave bioluminescence at 564 nm which is similar to LH2 at 561 nm. Further investigation by docking studies revealed that the emission wavelength of enamine luciferin might be attributed to the unwanted twisted structure caused by Asp531 within the enzyme. With mutant luciferase FlucRed, the major emission peak was shifted to 606 nm, a distinct shoulder above 700 nm, and 21% of its spectrum located in the nIR range

The impact of factors beyond Breslow depth on predicting sentinel lymph node positivity in melanoma

BACKGROUND. In addition to Breslow depth, the authors previously described how increasing mitotic rate and decreasing age predicted sentinel lymph node (SLN) metastases in patients with melanoma. The objectives of the current study were to verify those previous results and to create a prediction model for the better selection of which patients with melanoma should undergo SLN biopsy. METHODS. The authors reviewed 1130 consecutive patients with melanoma in a prospective database who underwent successful SLN biopsy. After eliminating patients aged <16 years and patients who had melanomas that measured <1 mm, 910 remaining patients were reviewed for clinical and pathologic features and positive SLN status. Univariate association of patient and tumor characteristics with positive SLN status was explored by using standard logistic regression techniques, and the best multivariate model that predicted lymph node metastases was constructed by using a backward stepwise-elimination technique. RESULTS. The characteristics that were associated significantly with lymph node metastasis were angiolymphatic invasion, the absence of regression, increasing mitotic rate, satellitosis, ulceration, increasing Breslow depth, decreasing age, and location (trunk or lower extremity compared with upper extremity or head/neck). Previously reported interactions between mitotic rate and age and between Breslow depth and age were confirmed. The best multivariate model included patient age (linear), angiolymphatic invasion, the number of mitoses (linear), the interaction between patient age and the number of mitoses, Breslow depth (linear), the interaction between patient age and Breslow depth, and primary tumor location. CONCLUSIONS. Younger age, increasing mitotic rate (especially in younger patients), increasing Breslow depth (especially in older patients), angiolymphatic invasion, and trunk or lower extremity location of the primary tumor were associated with a greater likelihood of positive SLN status. The current results support the use of factors beyond Breslow depth to determine the risk of positive SLN status in patients with cutaneous melanoma. Cancer 2007. © 2006 American Cancer SocietyPeer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/55870/1/22382_ftp.pd

What is a sentinel node? Re-evaluating the 10% rule for sentinel lymph node biopsy in melanoma

Introduction Many surgeons use the “10% rule” to define whether a lymph node is a sentinel node (SLN) when staging malignant melanoma. However, this increases the number of SLN removed and the time and cost of the procedure. We examined the impact of raising this threshold on the accuracy of the procedure. Methods We reviewed the records of 561 patients with melanoma (624 basins) who underwent SLN with technetium Tc99 labeled sulfur colloid using a definition of a SLN as 10% of that of the node with the highest counts per minute (CPM). Results Of the 624 basins, 154 (25%) were positive for metastases. An average of 1.9 nodes per basin were removed (range 1–6). Metastases were found in the hottest node in 137 cases (89% of positive basins, 97% of basins overall). Increasing the threshold above 10% decreased the number of nodes excised and the costs involved, but incrementally raised the number of false negative cases above baseline (a 4% increase for a “20% rule,” 5% for a “30% rule,” 6% for a “40% rule,” and 7% for a “50% rule”). Taking only the hottest node would raise the false negative rate by 11%. Conclusions Although using thresholds higher than 10% for the definition of a SLN will minimize the extent of surgery and decrease the costs associated with the procedure, it will compromise the accuracy of the procedure and is not recommended. J. Surg. Oncol. 2007;95:623–628. © 2007 Wiley-Liss, Inc.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/56055/1/20729_ftp.pd

A CANDELS - 3D-HST Synergy: Resolved Star Formation Patterns at 0.7 < z < 1.5

We analyze the resolved stellar populations of 473 massive star-forming galaxies at 0.7 < z < 1.5, with multi-wavelength broad-band imaging from CANDELS and Halpha surface brightness profiles at the same kiloparsec resolution from 3D-HST. Together, this unique data set sheds light on how the assembled stellar mass is distributed within galaxies, and where new stars are being formed. We find the Halpha morphologies to resemble more closely those observed in the ACS I band than in the WFC3 H band, especially for the larger systems. We next derive a novel prescription for Halpha dust corrections, which accounts for extra extinction towards HII regions. The prescription leads to consistent SFR estimates and reproduces the observed relation between the Halpha/UV luminosity ratio and visual extinction, both on a pixel-by-pixel and on a galaxy-integrated level. We find the surface density of star formation to correlate with the surface density of assembled stellar mass for spatially resolved regions within galaxies, akin to the so-called 'main sequence of star formation' established on a galaxy-integrated level. Deviations from this relation towards lower equivalent widths are found in the inner regions of galaxies. Clumps and spiral features, on the other hand, are associated with enhanced Halpha equivalent widths, bluer colors, and higher specific star formation rates compared to the underlying disk. Their Halpha/UV luminosity ratio is lower than that of the underlying disk, suggesting the ACS clump selection preferentially picks up those regions of elevated star formation activity that are the least obscured by dust. Our analysis emphasizes that monochromatic studies of galaxy structure can be severely limited by mass-to-light ratio variations due to dust and spatially inhomogeneous star formation histories.Comment: Accepted by The Astrophysical Journal, 18 pages, 1 table, 10 figure

3D-HST+CANDELS : the evolution of the galaxy size-mass distribution since z=3

Spectroscopic+photometric redshifts, stellar mass estimates, and rest-frame colors from the 3D-HST survey are combined with structural parameter measurements from CANDELS imaging to determine the galaxy size-mass distribution over the redshift range 0 < z < 3. Separating early- and late-type galaxies on the basis of star-formation activity, we confirm that early-type galaxies are on average smaller than late-type galaxies at all redshifts, and we find a significantly different rate of average size evolution at fixed galaxy mass, with fast evolution for the early-type population, R eff∝(1 + z)–1.48, and moderate evolution for the late-type population, R eff∝(1 + z)-0.75Peer reviewedFinal Accepted Versio

Rapid dissemination of Francisella tularensis and the effect of route of infection

<p>Abstract</p> <p>Background</p> <p><it>Francisella tularensis </it>subsp. <it>tularensis </it>is classified as a Category A bioweapon that is capable of establishing a lethal infection in humans upon inhalation of very few organisms. However, the virulence mechanisms of this organism are not well characterized. <it>Francisella tularensis </it>subsp. <it>novicida</it>, which is an equally virulent subspecies in mice, was used in concert with a microPET scanner to better understand its temporal dissemination in vivo upon intranasal infection and how such dissemination compares with other routes of infection. Adult mice were inoculated intranasally with <it>F. tularensis </it>subsp. <it>novicida </it>radiolabeled with ⁶⁴Cu and imaged by microPET at 0.25, 2 and 20 hours post-infection.</p> <p>Results</p> <p>⁶⁴Cu labeled <it>F. tularensis </it>subsp. <it>novicida </it>administered intranasally or intratracheally were visualized in the respiratory tract and stomach at 0.25 hours post infection. By 20 hours, there was significant tropism to the lung compared with other tissues. In contrast, the images of radiolabeled <it>F. tularensis </it>subsp. <it>novicida </it>when administered intragastrically, intradermally, intraperitoneally and intravenouslly were more generally limited to the gastrointestinal system, site of inoculation, liver and spleen respectively. MicroPET images correlated with the biodistribution of isotope and bacterial burdens in analyzed tissues.</p> <p>Conclusion</p> <p>Our findings suggest that Francisella has a differential tissue tropism depending on the route of entry and that the virulence of Francisella by the pulmonary route is associated with a rapid bacteremia and an early preferential tropism to the lung. In addition, the use of the microPET device allowed us to identify the cecum as a novel site of colonization of <it>Francisella tularensis </it>subsp. <it>novicida </it>in mice.</p

Deficiency and Also Transgenic Overexpression of Timp-3 Both Lead to Compromised Bone Mass and Architecture In Vivo

Tissue inhibitor of metalloproteinases-3 (TIMP-3) regulates extracellular matrix via its inhibition of matrix metalloproteinases and membrane-bound sheddases. Timp-3 is expressed at multiple sites of extensive tissue remodelling. This extends to bone where its role, however, remains largely unresolved. In this study, we have used Micro-CT to assess bone mass and architecture, histological and histochemical evaluation to characterise the skeletal phenotype of Timp-3 KO mice and have complemented this by also examining similar indices in mice harbouring a Timp-3 transgene driven via a Col-2a-driven promoter to specifically target overexpression to chondrocytes. Our data show that Timp-3 deficiency compromises tibial bone mass and structure in both cortical and trabecular compartments, with corresponding increases in osteoclasts. Transgenic overexpression also generates defects in tibial structure predominantly in the cortical bone along the entire shaft without significant increases in osteoclasts. These alterations in cortical mass significantly compromise predicted tibial load-bearing resistance to torsion in both genotypes. Neither Timp-3 KO nor transgenic mouse growth plates are significantly affected. The impact of Timp-3 deficiency and of transgenic overexpression extends to produce modification in craniofacial bones of both endochondral and intramembranous origins. These data indicate that the levels of Timp-3 are crucial in the attainment of functionally-appropriate bone mass and architecture and that this arises from chondrogenic and osteogenic lineages

The effects of pioglitazone, a PPARγ receptor agonist, on the abuse liability of oxycodone among nondependent opioid users

Aims: Activation of PPARγ by pioglitazone (PIO) has shown some efficacy in attenuating addictive-like responses in laboratory animals. The ability of PIO to alter the effects of opioids in humans has not been characterized in a controlled laboratory setting. The proposed investigation sought to examine the effects of PIO on the subjective, analgesic, physiological and cognitive effects of oxycodone (OXY). Methods: During this investigation, nondependent prescription opioid abusers (N = 17 completers) were maintained for 2-3 weeks on ascending daily doses of PIO (0 mg, 15 mg, 45 mg) prior to completing a laboratory session assessing the aforementioned effects of OXY [using a within-session cumulative dosing procedure (0, 10, and 20 mg, cumulative dose = 30 mg)]. Results: OXY produced typical mu opioid agonist effects: miosis, decreased pain perception, and decreased respiratory rate. OXY also produced dose-dependent increases in positive subjective responses. Yet, ratings such as: drug "liking," "high," and "good drug effect," were not significantly altered as a function of PIO maintenance dose. Discussion: These data suggest that PIO may not be useful for reducing the abuse liability of OXY. These data were obtained with a sample of nondependent opioid users and therefore may not be applicable to dependent populations or to other opioids. Although PIO failed to alter the abuse liability of OXY, the interaction between glia and opioid receptors is not well understood so the possibility remains that medications that interact with glia in other ways may show more promise

Nativity Differences in Stress among Asian and Pacific Islander American Women

According to the Stress Process Theory, people who are marginalized in society encounter more stress than those in more advantaged positions. Immigrants are one such marginalized group in the United States (US) who may experience greater psychological stress than their US-born counterparts due to (1) severing of social ties; (2) social disadvantage and marginalization; and (3) adaptation to a new environment. This study examines the disparity in stress by nativity, and how social factors contribute to this disparity for Asian and Pacific Islander (API) women. Data come from the Asian Community Health Initiative, which included a sample of 291 foreign-born and 155 US-born API women in the San Francisco Bay Area. Multivariable linear regression was used to estimate associations between nativity status and stress, measured using the Cohen Perceived Stress Scale, accounting for various social stressors. Foreign-born women had higher levels of stress compared to US-born. Stress was greater among women experiencing fewer socioeconomic resources, more discrimination, more acculturative stress, and low English proficiency. English proficiency accounted for much of the disparity in stress between foreign-born and US-born API women. This study contributes to our understanding of how stress among APIs is influenced by social disadvantage and marginalization in US society. Future research should further study how aspects of the immigrant experience are associated with stress among APIs over time