A Survey of Proteomic Biomarkers for Heterotopic Ossification in Blood Serum

Background: Heterotopic ossification (HO) is a significant problem for wounded warriors surviving high-energy blast injuries; however, currently, there is no biomarker panel capable of globally characterizing, diagnosing, and monitoring HO progression. The aim of this study was to identify biomarkers for HO using proteomic techniques and blood serum. Methods: Isobaric tags for relative and absolute quantitation (iTRAQ) was used to generate a semi-quantitative global proteomics survey of serum from patients with and without heterotopic ossification. Leveraging the iTRAQ data, a targeted selection reaction monitoring mass spectrometry (SRM-MS) assay was developed for 10 protein candidates: alkaline phosphatase, osteocalcin, alpha-2 type I collagen, collagen alpha-1(V) chain isoform 2 preprotein, bone sialoprotein 2, phosphatidate phosphatase LPIN2, osteomodulin, protein phosphatase 1J, and RRP12-like protein. Results: The proteomic survey of serum from both healthy and disease patients includes 1220 proteins and was enriched for proteins involved in the response to elevated platelet Ca+2, wound healing, and extracellular matrix organization. Proteolytic peptides from three of the ten SRM-MS proteins, osteocalcin preprotein, osteomodulin precursor, and collagen alpha-1(v) chain isoform 2 preprotein from serum, are potential clinical biomarkers for HO. Conclusions: This study is the first reported SRM-MS analysis of serum from individuals with and without heterotopic ossification, and differences in the serum proteomic profile between healthy and diseased subjects were identified. Furthermore, our results indicate that normal wound healing signals can impact the ability to identify biomarkers, and a multi-protein panel assay, including osteocalcin preproprotein, osteomodulin precursor, and collagen alpha-1(v) chain isoform 2 preprotein, may provide a solution for HO detection and monitoring

Reproductive Biology and Embryonic Diapause as a Survival Strategy for the East Asian Endemic Eagle Ray Aetobatus narutobiei

Batoids comprise five of the seven most threatened families of sharks and rays. The East Asian endemic Naru eagle ray Aetobatus narutobiei is a large bodied ray whose estuarine habitat overlaps with an economically valuable bivalve fishery. In response to decreased bivalve yields, the government initiated a predator control program and as a result, Naru eagle rays have faced intense and targeted fishing pressure during the last two decades. The long-term impacts of the predator control program on the population of rays and bivalves and their balance in the ecosystem are unknown because the life history of the Naru eagle ray has not been characterized. To begin to fill these critical knowledge gaps, the reproductive life history of the Naru eagle was described. Females mature at a larger size than males and require nearly twice as many years to reach maturity (DW50, 952.0 mm vs. 764.2 mm; Age50, 6.0 years vs. 3.5 years). Both males and females reproduce annually and their reproductive cycles are synchronized and seasonal. Females have a single ovary and paired uteri, are viviparous, and reproduce via matrotrophic histotrophy. Mating occurs in August and September and gestation lasts approximately 12 months including a 9.5-month diapause that begins soon after mating and ends in June of the following year, leaving 2.5 months for embryos to complete development. Fecundity ranged from 1 to 7 embryos per brood (n = 158, mean ± SD = 3.36 ± 1.26) and was positively correlated with female disc width (linear regression; F = 105.73, d.f. = 151, P < 0.05). Naru eagle rays are vulnerable to overfishing because of their low fecundity, long reproductive cycle and long time to reach sexual maturity. Obligate embryonic diapause during overwintering and seasonal migrations is a survival strategy that benefits the adults and neonates. This research is a valuable resource to help guide science-based management, conservation and protection of the endemic Asian A. narutobiei and its nursery areas

Monitoring egg fertility, embryonic morbidity, and mortality in an oviparous elasmobranch using ultrasonography

Ultrasonography is widely used to monitor pregnancy in viviparous species, but it is underutilized as a tool to characterize embryonic development in oviparous species. Currently, a multi-institutional effort is underway to re-wild the endangered zebra shark (Stegostoma tigrinum) to locations where this species was previously extirpated by leveraging the reproductive efforts of aquarium sharks as a source of brood stock. Zebra sharks are oviparous and fecund, but a large percentage of their yolked eggs do not result in hatchlings. Therefore, ultrasonography represents a potential tool for distinguishing fertile eggs with developing embryos from degrading eggs, and to diagnose changes in early embryonic development predictive of poor outcomes. The objectives of the current study were to use ultrasonography to assess egg fertility, monitor early embryonic development, and identify morphological indicators that may be predictive of early embryonic mortality. Freshly laid eggs from four female zebra sharks were collected and inventoried daily at Aquarium of the Pacific. Eggs were incubated undisturbed for 2 to 4 weeks and subsequently examined weekly via ultrasound to assess fertility and monitor embryo development. Among 120 fertile eggs, embryos were identified as early as 8 days post-oviposition, with average (±SD) time to first observation at 30 ± 7 days. Morphological and behavioral abnormalities were observed for most embryos (n = 84, 70%) as early as 16 days and up to 95 days post-oviposition. Common abnormalities included: bent or curled tails, vesicle(s) at the base of the yolk stalk, and slow or weak movement. Only one embryo survived to hatch during the study and was genetically-confirmed parthenogenetic, suggesting hatching success for parthenotes is low (&lt;1%). Ultrasonography was demonstrated to be an effective and non-invasive method to determine egg fertility, identify embryos with developmental abnormalities, and monitor embryo growth

Position of Insurance Broker in Insurance Market

Import 22/07/2015Abstrakt Jméno studenta: Bc. Klaudia Mičová Název práce: Postavení pojišťovacího makléře na pojistném trhu Práce se zabývá postavením pojišťovacího makléře na pojistném trhu. V teoretické části je vymezena základní charakteristika zprostředkovatelské činnosti, například rozdělení pojišťovacích zprostředkovatelů podle kategorií, registr, formy odměňování atd. Pojišťovací makléř jde zde podrobně popsán od povinností až po etický kodex. V praktické části dochází ke zhodnocení postavení pojišťovacího makléře na pojistném trhu jak ze strany klienta, tak ze strany pojišťovny.This work deals with the position of an insurance broker in the insurance market. In the theoretical part is defined the basic characteristic of brokering activity, such as the division of insurance brokers by category, register, forms of remuneration etc. Insurance Broker is described in detail in terms of obligations to the Code of Ethics. The practical part assesses the status of the insurance broker in the insurance market from both sides - the client and the insurance company.119 - Katedra právavelmi dobř

Granulation tissue of chronic pressure ulcers as a predictive indicator of wound closure.

: To describe the temporal relationship between the quantity of granulation tissue in a chronic pressure ulcer (PrU) and its clinical outcome. : Study participants were seen on days 0, 1, 2, 3, 4, 7, 8, 9, 10, 11, 14, 21, 28, 35, and 42. On each visit, the wounds were digitally photographed with a 3-cm calibration target. Images were analyzed using VeV MD (version 1.1.14; VERG Inc, Winnipeg, Manitoba, Canada) and Adobe Photoshop CS3 Extended (version 10.0.1; Adobe Systems Inc, San Jose, California). Granulation tissue was selected from calibrated digital images by 1 of 2 methods: manual selection and automated selection. Granulation tissue area was expressed as a percentage of total wound area. : Academic research laboratory. : Thirty-one chronic PrUs were observed in 27 subjects. : Quantitative measure of granulation tissue area. : There was no relationship between the amount of granulation tissue expressed as a percentage of the total PrU area and wound outcome. : This study is the first to both quantitatively measure the amount of granulation tissue in a chronic PrU and attempt to correlate it to wound outcome. Although counterintuitive, the amount of granulation tissue was not predictive of outcome, and no temporal trends could be described

Longitudinal study of stage III and stage IV pressure ulcer area and perimeter as healing parameters to predict wound closure

Documentation of healing progress is central to the plan of care for patients with a pressure ulcer. Several studies have shown that a reduction in wound area is a predictor of chronic wound healing, but data about pressure ulcers are limited. Furthermore, consensus is lacking as to which wound characteristics such as volume, area, and perimeter should be measured and what methods or tools to use when collecting measurements. This hampers comparisons among research studies and their eventual translation into clinical practice. The purpose of this longitudinal, repeated measurements study was to calculate healing parameters using wound area and perimeter measurements and evaluate their potential to predict closure. Twenty-seven (27) patients with 31 Stage III and Stage IV pressure ulcers participated in the 42-day study. Wound length, width, and perimeter were measured at 15 time points or until healing, and the following healing parameters were calculated: absolute area, percent area reduction, mean percent area reduction, trajectory, and three variations of the linear healing parameter. Ulcer size at day 0 was a significant predictor of time to heal (P = 0.0231). Smaller wounds required less time, but initial size did not influence wound outcome (P = 0.3537). Among ulcers that closed 81% or more of their original area, the initial linear healing rate (4 weeks) was 0.16 ± 0.02 cm/week and mean percent area reduction was 35.37% + 4.83, compared to 0.021 + 0.02 cm/week and - 4.66% + 6.24, respectively, for ulcers that closed 40% or less of their original area. Percent area reduction and linear healing parameters all were predictive of wound outcomes. Percent area measurements are easiest to determine but sensitive to initial wound size. The linear healing parameter requires calculation of both wound area and perimeter, but it is independent of initial wound size and yields rates directly comparable among wounds. These findings confirm that change in wound size after 4 weeks of care is a predictor of healing Stage III and Stage IV pressure ulcers. Future research studies should include other wound characteristics and patient comorbidities to further refine acceptable rates of wound closure

Fat Deposition and Fat Effects on Meat Quality—A Review

Growth is frequently described as weight gain over time. Researchers have used this information in equations to predict carcass composition and estimate fat deposition. Diet, species, breed, and gender all influence fat deposition. Alterations in diets result in changes in fat deposition as well as the fatty acid profile of meat. Additionally, the amount and composition of the fat can affect lipid stability and flavor development upon cooking. Fat functions not only as a storage of energy and contributor of flavor compounds, but also participates in signaling that affects many aspects of the physiological functions of the animal. Transcription factors that are upregulated in response to excess energy to be stored are an important avenue of research to improve the understanding of fat deposition and thus, the efficiency of production. Additionally, further study of the inflammation associated with increased fat depots may lead to a better understanding of finishing animals, production efficiency, and overall health

Ángel González Palencia: Moros y cristianos en la España medieval. Estudios históricos-literarios. Tercera seria. Madrid, Instituto Antonio de Nebrija (CSIC), 1945. VI + 350 pàgs. + 8 làms. - Del "Lazarillo" a Quevedo. Estudios histórico-literarios. Cuarta seria. Madrid, Instituto Antonio de Nebrija (CSIC), 1946. VIII + 432 pàgs. + 9 làms. - Eruditos y libreros del siglo XVIII. Estudios histórico-literarios. Quinta serie. Madrid, Instituto Antonio de Nebrija (CSIC), 1948. VIII + 446 pàgs. + I quadre genealògic.

Heterotopic ossification (HO) refers to the abnormal formation of bone in soft tissue. Although some of the underlying processes of HO have been described, there are currently no clinical tests using validated biomarkers for predicting HO formation. As such, the diagnosis is made radiographically after HO has formed. To identify potential and novel biomarkers for HO, we used isobaric tags for relative and absolute quantitation (iTRAQ) and high-throughput antibody arrays to produce a semi-quantitative proteomics survey of serum and tissue from subjects with (HO+) and without (HO−) heterotopic ossification. The resulting data were then analyzed using a systems biology approach. We found that serum samples from subjects experiencing traumatic injuries with resulting HO have a different proteomic expression profile compared to those from the matched controls. Subsequent quantitative ELISA identified five blood serum proteins that were differentially regulated between the HO+ and HO− groups. Compared to HO− samples, the amount of insulin-like growth factor I (IGF1) was up-regulated in HO+ samples, whereas a lower amount of osteopontin (OPN), myeloperoxidase (MPO), runt-related transcription factor 2 (RUNX2), and growth differentiation factor 2 or bone morphogenetic protein 9 (BMP-9) was found in HO+ samples (Welch two sample t-test; P \u3c 0.05). These proteins, in combination with potential serum biomarkers previously reported, are key candidates for a serum diagnostic panel that may enable early detection of HO prior to radiographic and clinical manifestations