From syndromes to normal variation: a candidate gene study of interorbital distances

Hypertelorism and telecanthus are clinical phenotypes associated with many genetic syndromes. To date, research is limited regarding whether disease-causing genes are related to normal craniofacial development in unaffected individuals. The aim of this study is to determine whether common genetic variation in forty selected genes implicated in hypertelorism/telecanthus-related syndromes contribute to normal variation of intercanthal and outer-canthal distances of the orbits. Hypertelorism/telecanthus-related genes were selected based on significant prevalence of the phenotype in the described genetic syndrome. Using the 3D Facial Norms (TDFN) Repository, genomic and anthropometric data were utilized to test genetic association for common variants in two phenotypes: intercanthal and outer-canthal distances. Suggestive SNPs with evidence of association were annotated for relevant gene function related to craniofacial development. For the intercanthal distance measurement, one statistically significant SNP (p<4.05x10-6) in LINC00482 and two suggestive SNPs (p<10-4), one in HMGCS2 and another within 200kB of FAM58A, were observed. For the outer-canthal distance measurement, five suggestive SNPs (p<10-4) were observed near ADAMTS18, GLI3, ACTG1, MEGF11, and SPECC1L. We hypothesize that identified SNPs have regulatory effects on the expression of these genes and contribute to interorbital distances in unaffected individuals. Identifying genetic determinants of craniofacial development in the normal population is important for the understanding of mechanisms underlying craniofacial dysmorphology. In addition, understanding the mechanisms that contribute to the transition from normal variation to a disease state in a population is important to public health because most genetic diseases exist on a spectrum. With better understanding the unaffected side of the spectrum allows us to better identify the disease side of the spectrum, allowing for better diagnosis and treatment for individuals with craniofacial anomalies. This study attempts to identify these risk loci and hypothesize what impact these loci might have on craniofacial development

Exogenous influences on skatole formation in the pig

Castration of male piglets without anesthesia and analgesia has been an ancient and traditional practice in several European countries to prevent problems with boar taint. The commencement of the National Protection of Animals Act on 13th July 2013 intends to end castration without anesthesia in Germany by 2019. In Europe, the goal is to end castration of piglets by raising entire male pigs only from 2018 onwards. This, however, is a challenge for the entire supply chain of the pig production industry, since sufficient consumer protection from tainted boar meat cannot be guaranteed so far. Off-odors in boar meat are caused by the testicular steroid androstenone and the tryptophane metabolite skatole. Exogenous and endogenous factors which favor the formation of both boar taint substances, but especially of skatole, have so far been only partially identified. Thus the aim of the present thesis was to determine exogenous influences on skatole formation and deduce measures to reduce the amount of tainted carcasses. In the first part of the thesis (Chapter 3), the current state of research on the impact of feeding strategies on skatole physiology was summarized. The mechanisms of different feeding strategies and feed additives were described in their effect on formation, metabolism and fat deposition of skatole. Based on a deduced formation cascade of skatole, different feeding strategies aiming to reduce skatole can be evaluated in a simplified manner. It could be shown that promising feeding strategies to reduce skatole have to be effective on more than one level of the formation cascade at the same time. In the second part of this thesis (Chapter 4), the reasons for varying androstenone and skatole concentrations found at slaughter plants in animals from the same origin were investigated. In order to identify the reasons leading to said differences, a study with 169 boars from three different farms was conducted. Each farm delivered animals, split into two groups, to two different slaughter plants with a time interval of one day. The duration of transport as well as the duration of pre-unloading and the time spent on the vehicle before unloading were recorded. During the slaughter process, samples of blood, feces and urine were collected for hormone analysis. Carcasses were scored visually for lesions in cold storage after slaughter, and adipose tissue was removed for boar taint analysis. Even in animals of the same origin, the same genotype as well as the same feeding regimen, significant differences in androstenone and skatole concentrations in fat could be measured, which could be traced back to the different transport and pre-unloading times. In contrast to androstenone, skatole in the fat of the animals was influenced predominately by the pre-unloading time and increased by more than 20 ng/g with every hour of pre-unloading time. In addition, animals with higher lesion scores had higher skatole concentrations in fat. Transport time, on the other hand, had an effect on androstenone concentrations in fat, which increased by about 0.1 µg/g per hour transport. Skatole concentrations, however, were only slightly affected by transport time. Positive correlations could be found between cortisol and testosterone in various substrates with deposition of boar taint substances in fat. However, further research is required to clarify the mechanisms of these effects in detail. Minimizing transport and pre-unloading times before the slaughter process, however, seems mandatory to reduce the amount of tainted carcasses. The third part of this thesis (Chapter 5) investigated the impact of management factors on variability of skatole concentrations in blood and fat. Modern breeding companies take fat biopsy samples to estimate the breeding value for the trait boar taint in AI boars. However, it is not yet known to which extent repeated biopsies or different sampling locations may themselves affect skatole levels. Furthermore, the influence skin contamination of animals and of transdermal skatole diffusion have been matters of heated discussion for decades. The published results, however, are in part contradictory and it is not clear to which extent skatole can diffuse through the skin and to which degree this source may contribute to the concentrations in the carcass. As a consequence, the clarification of these complex relationships was the subject of the third study. The results show that skatole concentrations reveal a low variability in samples from the dorsal part of the carcass, although 20% higher concentrations were measured in the ventral area. The transdermal diffusion of skatole was confirmed in this study, but it was also shown that this diffusion is local and that skatole levels of the carcass in general were not increased. Repeated biopsies under total anesthesia temporarily increased skatole levels in blood and were accompanied by a similar course of cortisol concentrations in blood. Punch biopsies in conscious animals had no effect on the course of skatole or cortisol in blood. The present thesis illustrates that, besides nutrition, stress is an important factor for off-odors in boar carcasses. The results from the experimental studies suggest that an improvement of animal welfare can reduce the risk of off-odor in entire male pig production and thus contribute to consumer protection. However, the results also show that any success of the farmer in the reduction of boar taint can be partially undone on the way to slaughter.Die betäubungslose Kastration männlicher Ferkel ist seit Jahrhunderten in vielen europäischen Ländern übliche Praxis um Schlachtkörper mit Ebergeruch zu verhindern. Mit Inkrafttreten der Novellierung des Tierschutzgesetzes am 13. Juli 2013, wurde beschlossen bis zum Jahr 2019 in Deutschland aus der betäubungslosen Ferkelkastration auszusteigen. Europaweit ist man bestrebt bereits 2018 auf die Ferkelkastration gänzlich zu verzichten und Jungeber zu mästen. Dies stellt allerdings eine Herausforderung für die gesamte Kette der Schweinefleischerzeugung dar, da bisher ein ausreichender Schutz der Verbraucher vor geruchsbelastetem Fleisch nicht sichergestellt ist. Die Geruchsbelastungen von Eberfleisch werden durch das Hodensteroid Androstenon und den Tryptophanmetaboliten Skatol verursacht. Exogene und endogene Faktoren die die Bildung der beiden Ebergeruchstoffe - insbesondere von Skatol - begünstigen, sind allerdings nur zum Teil identifiziert. Das Hauptziel dieser Arbeit war es daher exogene Einflussgrößen auf die Bildung von Skatol zu untersuchen und hieraus Maßnahmen zur Verminderung des Anteils geruchsbelasteter Schlachtkörper abzuleiten. Im ersten Teil dieser Arbeit (Chapter 3) wurde der aktuelle Kenntnisstand zur Auswirkung von Fütterungsmaßnahmen auf die Skatolphysiologie zusammengefasst. Dabei wurden die Mechanismen beschrieben, über die verschiedene Fütterungsstrategien oder Futterzusätze in die Bildung, Metabolisierung und Einlagerung von Skatol ins Fett eingreifen. Anhand einer daraus abgeleiteten Bildungskaskade können skatolreduzierende Fütterungsmaßnahmen hinsichtlich ihrer Wirksamkeit vereinfacht beurteilt werden. Dabei konnte gezeigt werden, dass erfolgversprechende Fütterungsmaßnahmen zur Reduzierung von Skatol möglichst gleichzeitig auf mehreren Stufen der Bildungskaskade ihre Wirkung entfalten müssen. Im zweiten Teil dieser Arbeit (Chapter 4) wurde untersucht, warum Androstenon- und Skatolkonzentrationen bei Tieren gleicher Herkunft zwischen Schlachthöfen variieren können. Zur Identifikation der Ursachen wurde eine Studie an zwei verschiedenen Schlachthöfen durchgeführt, an denen 169 Eber aus drei verschieden Betrieben geschlachtet wurden Jeder Betrieb lieferte seine Tiere in zwei Gruppen an die beiden Schlachthöfe im Anstand von einem Tag an. Dabei wurden die Transportzeit und die Wartezeit im LKW vor dem Abladen am Schlachthof erfasst. Während des Schlachtvorgangs wurden Blut, Kot und Urin für die Hormonanalytik gesammelt. Im Kühlhaus nach dem Schlachten wurden die Schlachtkörperhälften nach Verletzungsspuren bonitiert und Fettgewebe für die Ebergeruchsbestimmung entnommen. Bei Tieren gleicher Herkunft, gleicher Genetik sowie gleicher Fütterung konnten Unterschiede in Androstenon- und Skatolwerten aufgezeigt werden, welche durch die unterschiedlichen Transport- und Wartezeiten erklärt werden konnten. Dabei wurde im Gegensatz zu Androstenon Skatol im Fett der Tiere maßgeblich von der Dauer der Wartezeit beeinflusst, je Stunde Wartezeit stiegen die Skatolkonzentrationen um mehr als 20 ng/g Fett an. Zudem wiesen Tiere mit höherem Verletzungsgrad auch höhere Skatolkonzentrationen auf. Die Transportzeit hatte hingegen Einfluss auf die Androstenonkonzentrationen, diese stiegen je Stunde Fahrt um etwa 0,1 µg/g Fett an, während die Skatoleinlagerung nur geringfügig beeinflusst wurde. Dabei konnten positive Beziehungen zwischen Cortisol und Testosteron in den verschiedenen Substraten und der Einlagerung geruchsaktiver Substanzen ins Fett nachgewiesen werden, allerdings sind weitere Untersuchungen notwendig um die detaillierten Mechanismen darstellen zu können. Eine Minimierung von Transport- und Wartezeit bei der Schlachtung von Jungebern scheint zwingend, um den Anteil geruchsbelasteter Schlachtkörper zu minimieren. Im dritten Teil dieser Arbeit (Chapter 5) wurde der Einfluss von Managementfaktoren auf die Variabilität von Skatolkonzentration in Blut und Fett untersucht. Moderne Zuchtunternehmen entnehmen Fettbiopsien am lebenden Schwein um die Eigenleistung für den Zuchtwert Ebergeruch potentieller Besamungseber bestimmen zu können. Es ist jedoch unklar inwieweit wiederholte Biopsien oder unterschiedliche Entnahmestellen die gemessenen Skatolkonzentrationen beeinflussen können. Zudem wird der Einfluss des Verschmutzungsgrades der Tiere und einer Diffusion von Skatol durch die Haut bereits seit langem diskutiert. Allerdings sind die publizierten Ergebnisse hierzu teilweise widersprüchlich und lassen die Frage offen, in welchem Maß Skatol es durch die Haut diffundieren kann und welchen Beitrag diese Quelle zu den Konzentrationen im Schlachtkörper leistet. Die Klärung dieser komplexen Zusammenhänge war Gegenstand der dritten Untersuchung. Die Ergebnisse zeigen, dass die Skatolkonzentration in Proben aus dem dorsalen Bereich eines Schlachtkörpers nur eine geringe Variation aufweisen, allerdings im ventralen Bereich um 20% höher sind. Die transdermale Diffusion von Skatol konnte prinzipiell bestätigt werden. Dabei konnte auch gezeigt werden, dass die Diffusion lokal begrenzt ist und nicht systemisch die Skatolkonzentrationen eines Schlachtkörpers erhöht. Wiederholte Biopsien unter Narkose erhöhen vorübergehend die Skatolkonzentrationen im Blut und sind von einem gleichsinnigen Verlauf der Cortisolkonzentrationen begleitet. Eine Schussbiopsie ohne Narkose zur Gewebenentnahme hatte keinen Effekt auf den Verlauf der Skatol- und Cortisolkonzentrationen im Blut. Die vorgelegte Arbeit zeigt, dass neben der Ernährung der Faktor Stress eine wichtige Einflußgröße auf die Geruchsbelastung von Eberschlachtkörpern ist. Die Ergebnisse des experimentellen Teils legen nahe, dass eine Verbesserung des Tierschutzes im Produktionsverfahren Ebermast das Risiko von Geruchsabweichungen bei Ebern vermindern kann und somit auch zum Verbraucherschutz beiträgt. Die Ergebnisse zeigen aber auch, dass auf dem Weg zur Schlachtung die Erfolge des Landwirts bei der Reduzierung des Ebergeruchs partiell wieder zunichte gemacht werden können

Role of Rad54, Rad54b and Snm1 in DNA damage repair

The aim of this thesis is to investigate the function of a number of genes involved in mammalian DNA damage repair, in particular in repair of DNA double-strand breaks (DSBs). Among a large number of different damages that can be introduced to DNA, DSBs are especially toxic. If left unrepaired, DSBs can trigger apoptosis or induce chromosomal rearrangements that can lead to carcinogenesis. Two main pathways are responsible for repair of DSBs: homologous recombination (HR) and nonhomologous end joining (NHEJ). HR is generally an error-free mechanism that restores missing information on the basis of homologous sequence obtained from sister chromatid or homologous chromosome. By contrast, NHEJ is generally error-prone. During repair by NHEJ the DNA ends can be directly ligated or short stretches of homology at the ends can be used, leading to deletions or insertions at the site of the break. A number of genes have been identified as players in DSB repair, both in prokaryotes and eukaryotes. Many of them are found in all the kingdoms of life and are similar in aspects of their sequence and function, although there are genes characteristic only for prokaryotes or eukaryotes. Many subtle differences in function also emerge from studies on HR proteins in different species. These differences might have appeared because of diverse functions repair genes have to perform in more complexed organisms, or because the initial function(s) of a gene has been distributed over multiple paralogues. Herein I concentrate on genes involved mainly in DSB repair via HR in mammalian systems. Two members of the group of HR genes are studied: Rad54 and its paralogue Rad548. Using mice and cells deficient in these genes we try to define the role of both Rad54 and Rad54B in HR and their contribution to other cellular processes. Additionally, we investigate the link between Rad54 and Snm1, a gene originally identified as being important for interstrand cross link (ICL) repair, with regard to ionizing radiation-induced DNA damage repair

Quantitative analysis of active pharmaceutical ingredients (APIs) using a potentiometric electronic tongue in a SIA flow system

Research funding: National Science Centre. Grant Numbers: DEC-2013/09/B/ST4/00957, LIDER/17/202/L-1/09/NCBiR/2010An advanced potentiometric electronic tongue and Sequential Injection Analysis (SIA) measurement system was applied for the quantitative analysis of mixtures containing three active pharmaceutical ingredients (APIs): acetaminophen, ascorbic acid and acetylsalicylic acid, in the presence of various amounts of caffeine as interferent. The flow-through sensor array was composed of miniaturized classical ion-selective electrodes based on plasticized PVC membranes containing only ion exchangers. Partial Least Squares (PLS) analysis of the steady-state sensor array responses, measured in API mixtures prepared by the SIA system permitted a correct quantitative analysis of acetylsalicylic acid and ascorbic acid. Further optimization using multiway PLS fed by dynamic responses without additional feature extraction did not improve significantly the resolution of acetaminophen. Lastly, the chemometric treatment, involving the extraction of dynamic components of the transient response employing the Wavelet transform, the removal of less-significant coefficients by means of Causal Index pruning and training of an Artificial Neural Network (ANN) with the selected coefficients, allowed the simultaneous determination of all the three studied APIs, while counterbalancing any interference due to caffeine

The epigenetic landscape of clear-cell renal cell carcinoma

Clear cell renal cell carcinoma (ccRCC) is the most common subtype of all kidney tumors. During the last few years, epigenetics has emerged as an important mechanism in ccRCC pathogenesis. Recent reports, involving large-scale methylation and sequencing analyses, have identified genes frequently inactivated by promoter methylation and recurrent mutations in genes encoding chromatin regulatory proteins. Interestingly, three of detected genes (PBRM1, SETD2 and BAP1) are located on chromosome 3p, near the VHL gene, inactivated in over 80% ccRCC cases. This suggests that 3p alterations are an essential part of ccRCC pathogenesis. Moreover, most of the proteins encoded by these genes cooperate in histone H3 modifications. The aim of this review is to summarize the latest discoveries shedding light on deregulation of chromatin machinery in ccRCC. Newly described ccRCC-specific epigenetic alterations could potentially serve as novel diagnostic and prognostic biomarkers and become an object of novel therapeutic strategies

Building Capacity for Volunteerism and Social Support at Leeway, Inc., a Skilled Nursing Facility

Leeway Inc., a skilled nursing center in New Haven that cares for individuals with HIV/AIDS, was founded in 1995. Its continuum of care includes skilled nursing care, residential care, independent housing, and community case management. It provides a wide variety of care, which includes medical, behavioral, and nursing services, as well as mental health services and addiction treatment. One of Leeway’s priorities is to address the psychiatric disorders that many HIV/AIDS patients have, such as depression, anxiety, addiction, and substance-related disorders (Leeway, 2012). Among patients admitted to Leeway between October 1995 and December 1999, 25% had severe, long-term comorbid psychiatric disorders. Each year, the prevalence of psychiatric comorbid disorders has increased. Between 1996 and 1998, the prevalence of psychiatric comorbid disorders increased from 38% to 53%. This trend could be due to low discharge rates and longer lengths of stay among these individuals (Goulet, 2000). Leeway places a significant emphasis on creating a plan of care that addresses and manages these disorders (Leeway, 2012). Some of the obstacles faced by the residents include social stigma, lack of engagement in their community, and lack of companionship. Moreover, the resident population is unique for several reasons, one of which is age. Leeway residents are younger than those at the typical nursing home facility, which introduces a variety of challenges to keep them engaged in activities. A current volunteer program has attempted to address many of these obstacles, but demonstrates the need for improvement. Residents reported differing levels of interest in both the frequency and structure of volunteer engagement. Staff also expressed the needs to balance the benefits and drawbacks of an individualized program, a “buddy” or more group activities facilitated by volunteers which would help increase companionship and engagement among residents. Residents also frequently expressed satisfaction with activities outside of the facility, which is one way in which the program could be expanded in the future. Recommendations and Products: 1. Advertise the volunteer positions as “Community Programming Volunteers” to boost enrollment and encourage a sense of agency and investment, 2. Hire a volunteer coordinator to serve as a point of contact, summarize feedback for staff, lead new initiatives, and perform basic administrative tasks, 3. Divide the year into three terms, and hold one orientation session at the beginning of each term, 4. Replace the ad hoc system of volunteer applications, training, and feedback with concise online platforms that automatically feed into a central system of records, 5. Create a separate but converging program structure that allows for two screening and orientation processes for regular and sporadic volunteers, 6. Create a two-part schedule for volunteer sessions: a. Matching interested residents with regular volunteers based on interests. Residents and volunteer buddies will meet for a 10-20 minute check-in with the option of joining the group activities and b. Regular and sporadic volunteers engage in a group activity or game, 7. Implement a hybrid individualized-group volunteer program structure, 8. Provide volunteers with nametags, and 9. Connect with schools and colleges throughout New Haven to access new volunteer pools.https://elischolar.library.yale.edu/ysph_pbchrr/1050/thumbnail.jp

Signal Integration of IFN-I and IFN-II With TLR4 Involves Sequential Recruitment of STAT1-Complexes and NFkB to Enhance Pro-inflammatory Transcription

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Independent comparison study of six different electronic tongues applied for pharmaceutical analysis

Electronic tongue technology based on arrays of cross-sensitive chemical sensors and chemometric data processing has attracted a lot of researchers' attention through the last years. Several so far reported applications dealing with pharmaceutical related tasks employed different e-tongue systems to address different objectives. In this situation, it is hard to judge on the benefits and drawbacks of particular e-tongue implementations for R&D in pharmaceutics. The objective of this study was to compare the performance of six different e-tongues applied to the same set of pharmaceutical samples. For this purpose, two commercially available systems (from Insent and AlphaMOS) and four laboratory prototype systems (two potentiometric systems from Warsaw operating in flow and static modes, one potentiometric system from St. Petersburg, one voltammetric system from Barcelona) were employed. The sample set addressed in the study comprised nine different formulations based on caffeine citrate, lactose monohydrate, maltodextrine, saccharin sodium and citric acid in various combinations. To provide for the fair and unbiased comparison, samples were evaluated under blind conditions and data processing from all the systems was performed in a uniform way. Different mathematical methods were applied to judge on similarity of the e-tongues response from the samples. These were principal component analysis (PCA), RV' matrix correlation coefficients and Tuckeŕs congruency coefficients

Genome-wide inhibition of pro-atherogenic gene expression by multi-STAT targeting compounds as a novel treatment strategy of CVDs

Cardiovascular diseases (CVDs), including atherosclerosis, are globally the leading cause of death. Key factors contributing to onset and progression of atherosclerosis include the pro-inflammatory cytokines Interferon (IFN)a and IFN? and the Pattern Recognition Receptor (PRR) Toll-like receptor 4 (TLR4). Together, they trigger activation of Signal Transducer and Activator of Transcription (STAT)s. Searches for compounds targeting the pTyr-SH2 interaction area of STAT3, yielded many small molecules, including STATTIC and STX-0119. However, many of these inhibitors do not seem STAT3-specific. We hypothesized that multi-STAT-inhibitors that simultaneously block STAT1, STAT2, and STAT3 activity and pro-inflammatory target gene expression may be a promising strategy to treat CVDs. Using comparative in silico docking of multiple STAT-SH2 models on multi-million compound libraries, we identified the novel multi-STAT inhibitor, C01L-F03. This compound targets the SH2 domain of STAT1, STAT2, and STAT3 with the same affinity and simultaneously blocks their activity and expression of multiple STAT-target genes in HMECs in response to IFNa. The same in silico and in vitro multi-STAT inhibiting capacity was shown for STATTIC and STX-0119. Moreover, C01L-F03, STATTIC and STX-0119 were also able to affect genome-wide interactions between IFN? and TLR4 by commonly inhibiting pro-inflammatory and pro-atherogenic gene expression directed by cooperative involvement of STATs with IRFs and/or NF-κB. Moreover, we observed that multi-STAT inhibitors could be used to inhibit IFN?+LPS-induced HMECs migration, leukocyte adhesion to ECs as well as impairment of mesenteric artery contractility. Together, this implicates that application of a multi-STAT inhibitory strategy could provide great promise for the treatment of CVDsThis publication was supported by grants UMO-2015/17/B/NZ2/00967 (HB) and UMO-2015/16/T/NZ2/00055 (MS) from National Science Centre Poland. This work was supported by the KNOW RNA Research Centre in Poznan (No. 01/KNOW2/2014) and in part by PL-Grid Infrastructure (MS