Cord blood adipokines and lipids and adolescent nonalcoholic fatty liver disease

© 2016 by the Endocrine Society. Context: Maternal adiposity in pregnancy is associated with offspring adiposity and metabolic dysfunction postnatally, including greater risk of nonalcoholic fatty liver disease (NAFLD). Recent genetic analyses suggest a causal effect of greater maternal body mass index on offspring birth weightandponderal index, but the relative roles of the environment in utero or later in life remains unclear. Objective: We sought to determine whether markers of infant adiposity (birth weight, umbilical cord blood leptin, adiponectin, and lipids) were associated with markers of NAFLD in adolescence. Design, Setting, and Participants: This was aUK prospective birth cohort with 17 years of follow-up with liver function tests (aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltransferase) (n = 1037 participants), and ultrasound scan assessed liver fat, volume, and sheer velocity at age 17 (n = 541 participants). Missing covariate data were imputed. Main Outcomes: Ultrasound and biochemical measures of NAFLD were measured. Results: Birth weight, cord blood leptin, and adiponectin were not associated with a diagnosis of NAFLD. In adjusted analyses, 2 of 42 associations attained conventional 5% levels of significance. Birth weight was positively associated with liver volume (1.0% greater per 100 g [95% confidence interval 0.5%-2.0%]). Cord high-density lipoprotein-cholesterol was positively associated with alanine aminotransferase (11.6% higher per 1 mmol/L [95% confidence interval 0.3, 23.4]); however, this association was primarily mediated via offspring adiposity. Conclusions: In this extensive analysis, we found little evidence measurements of infant fat mass and birth size were related to adolescent markers of NAFLD. The association between birth weight and adolescent liver volume may indicate the contribution of greater organ size to birth weight and tracking of organ size

Preclinical evaluation of a TEX101 protein ELISA test for the differential diagnosis of male infertility

BACKGROUND: TEX101 is a cell membrane protein exclusively expressed by testicular germ cells and shed into seminal plasma. We previously verified human TEX101 as a biomarker for the differential diagnosis of azoospermia, and developed a first-of-its-kind TEX101 ELISA. To demonstrate the clinical utility of TEX101, in this work we aimed at evaluating ELISA performance in a large population of fertile, subfertile, and infertile men. METHODS: Mass spectrometry, size-exclusion chromatography, ultracentrifugation, and immunohistochemistry were used to characterize TEX101 protein as an analyte in seminal plasma. Using the optimized protocol for seminal plasma pretreatment, TEX101 was measured by ELISA in 805 seminal plasma samples. RESULTS: We demonstrated that TEX101 was present in seminal plasma mostly in a free soluble form and that its small fraction was associated with seminal microvesicles. TEX101 median values were estimated in healthy, fertile pre-vasectomy men (5436 ng/mL, N = 64) and in patients with unexplained infertility (4967 ng/mL, N = 277), oligospermia (450 ng/mL, N = 270), and azoospermia (0.5 ng/mL, N = 137). Fertile post-vasectomy men (N = 57) and patients with Sertoli cell-only syndrome (N = 13) and obstructive azoospermia (N = 36) had undetectable levels of TEX101 (≤0.5 ng/mL). A cut-off value of 0.9 ng/mL provided 100% sensitivity at 100% specificity for distinguishing pre- and post-vasectomy men. The combination of a concentration of TEX101 > 0.9 ng/mL and epididymis-specific protein ECM1 > 2.3 μg/mL provided 81% sensitivity at 100% specificity for differentiating between non-obstructive and obstructive azoospermia, thus eliminating the majority of diagnostic testicular biopsies. In addition, a cut-off value of ≥0.6 ng/mL provided 73% sensitivity at 64% specificity for predicting sperm or spermatid retrieval in patients with non-obstructive azoospermia. CONCLUSIONS: We demonstrated the clinical utility of TEX101 ELISA as a test to evaluate vasectomy success, to stratify azoospermia forms, and to better select patients for sperm retrieval. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12916-017-0817-5) contains supplementary material, which is available to authorized users

Role of quorum sensing in adaptation of Bacillus subtilis to high salinity

Bacillus subtilis živi v okolju, kjer potekajo neprestane osmotske spremembe. V magistrski nalogi smo želeli ugotoviti vpliv sistema za zaznavo celične gostote oziroma kvoruma ComQXPA in z DegQ uravnanega sistema DegS-DegU na prilagajanje bakterije B. subtilis na povišano slanost. Vključili smo divji tip seva PS-216, mutanto PS-216 ΔcomQ, ki ne tvori aktivnega signala ComX sistema ComQXPA, in mutanto PS-216 ΔdegQ, ki ne tvori DegQ. Sevom smo določili rast in morfološke lastnosti kolonij v tekočih in na trdnih gojiščih SM in MSgg z različnimi koncentracijami NaCl. V nadaljevanju smo spremljali izražanje fluorescenčno označenih promotorjev genov proH, epsA in srfAA pri različnih slanostih tekočega gojišča MSgg. Primerljive velikosti površin nastalih kolonij pri mutanti ΔcomQ pri vseh slanostih in hitrejši začetek izražanja promotorja gena proH ob povišani slanosti v primerjavi z divjim tipom kažejo na boljšo prilagoditev mutante ΔcomQ na povišano slanost. Izražanje promotorja gena epsA je bilo pri mutanti ΔcomQ višje kot pri divjem tipu, kar kaže na boljšo tvorbo biofilma. Manjša aktivnost sistema DegS-DegU pri mutanti ΔdegQ vpliva na morfologijo kolonij, medtem ko je dinamika začetka tvorbe kolonij in njihova končna površina podobna kot pri divjem tipu. Promotorja genov proH in epsA se izražata pozneje in manj intenzivno, kar kaže na slabšo prilagoditev na povišano slanost ter inhibirano tvorbo biofilma. Izražanje promotorja gena srfAA je bilo najvišje pri mutanti ΔdegQ, sledil je divji tip in najnižjo aktivnost smo zasledili pri mutanti ΔcomQ. Iz pridobljenih rezultatov lahko sklepamo, da odsotnost sistema za zaznavanje kvoruma ComQXPA pospeši prilagoditev mutante ΔcomQ na povišano slanost, medtem ko manjša aktivnost sistema DegS-DegU prilagoditev mutante ΔdegQ na povišano slanost upočasni.Bacillus subtilis lives in an environment with frequent osmotic changes. In this master\u27s thesis we aimed to investigate the influence of the DegQ-controlled DegS-DegU system and ComQXPA system, involved in cell density or quorum sensing, on the adaptation of B. subtilis to high salinity. Strains included were PS-216 wild-type (wt) strain, mutant strain PS-216 ΔcomQ without production of an active signal ComX of the ComQXPA system, and mutant strain PS-216 ΔdegQ without DegQ production. Growth and morphological properties of colonies in liquid and solid SM and MSgg media with different NaCl concentrations were determined. In addition, the expression of fluorescently labelled proH, epsA, and srfAA gene promoters was monitored at different salt levels in MSgg liquid medium. Similar areas of the colonies of the mutant ΔcomQ at all salinities and faster beginning of the proH gene promoter expression at increased salinity in comparison to the wt strain show more efficient mechanism of adaptation of the mutant ΔcomQ strain to high salinity. Expression of the epsA gene promoter was higher in the ΔcomQ mutant than in the wt strain, indicating better biofilm formation. The lower activity of the DegS-DegU system in the ΔdegQ mutant affects colony morphology, while the dynamics of colony formation and their final size are similar to those of the wt strain. The promoters of proH and epsA genes are expressed later and less intensely, indicating poorer adaptation to increased salinity and inhibited biofilm formation. The highest expression of srfAA gene promoter was in the ΔdegQ mutant, followed by the wt and ΔcomQ mutant. From the results obtained, we can conclude that the absence of the ComQXPA system accelerates the adaptation of the ΔcomQ mutant to increased salinity, while the lower activity of the DegS-DegU system slows down the adaptation of the ΔdegQ mutant to increased salinity

Visual identity redesign of the Belokranjski museum in Metlika

V diplomskem delu sem oblikoval vizualno identiteto Belokranjskega muzeja Metlika. V teoretičnem delu sem raziskal pojem vizualne identitete in zgodovino njenega nastanka, raziskal in predstavil strukturo dinamične vizualne identitete, okoliščine in tehnologije, ki so vplivale na njen nastanek, in njen vpliv na naslovnika sporočila. Analitični del sestavljajo primeri dobrih praks oblikovanja za kulturne institucije, pri čemer sem predstavil vizualne identitete glasbenega centra Casa de Musica, muzeja sodobne umetnosti MoMA in filmskega inštituta Cinémathèque française. Sledi predstavitev Belokranjskega muzeja Metlika, njegove vsebine in poslanstva ter analiza njegove obstoječe vizualne identitete. V praktičnem delu sem oblikoval vizualno identiteto Belokranjskega muzeja Metlika. Predstavil sem osnovne gradnike vizualne identitete, konstrukcijo, pravila za načrtovanje in uporabo dinamičnega logotipa ter digitalne in tiskane aplikacije.For my thesis, I designed a visual identity of the Bela krajina Museum Metlika. In the theoretical part, I researched the concept of a visual identity, history of its creation, I presented the structure of a dynamic visual identity, the circumstances and technology that influenced its creation and its impact on the recipient of the message. In the analytical part of the thesis I researched some of the best practices of designing for cultural institutions. I presented the visual identities of the musical center Casa de Musica, the museum of contemporary art MoMA and the film institute Cinémathèque française. This is followed by a presentation of the Bela krajina Museum Metlika, its contents and mission, as well as an analysis of its existing visual identity. In the practical part, I designed the visual identity of the Bela krajina Museum Metlika. I presented the building blocks of the visual identity, construction, rules for designing and usage of the dynamic logo, as well as some digital and printed applications

Problems of penological treatment of juvenile and older minor offenders in SR Slovenia

Purpose of these research was to give an evaluation of influence on executing institutional measures on juvenile and older minor offenders in SR Slovenia. Starting point of these research was, that judicial system and system of executing punishment by juvenile offenders can, especially at the beginning, promote rise of criminal career (recidivism). The survey was conducted in two parts. The first part (F2) was carried out among employees in the Slovenian detention centers, the second part (F1) was carried out between those convicted in criminal Slovenian detention centers.Purpose of these research was to give an evaluation of influence on executing institutional measures on juvenile and older minor offenders in SR Slovenia. Starting point of these research was, that judicial system and system of executing punishment by juvenile offenders can, especially at the beginning, promote rise of criminal career (recidivism). The survey was conducted in two parts. The first part (F2) was carried out among employees in the Slovenian detention centers, the second part (F1) was carried out between those convicted in criminal Slovenian detention centers

IgG-mediated Immune Suppression: the Effect on the Host Immune System

One of the most effective immunological interventions for human disease prevention is the administration of anti-red blood cell (RBC) IgG, more specifically, anti-D IgG, for prevention of hemolytic disease of the fetus and newborn (HDN), a serious and potentially fatal condition caused by the maternal immune response against the Rhesus (Rh) blood group system D antigen on fetal RBC. Despite its widespread clinical use, the mechanism of the suppressive anti-RBC IgG effect is not fully understood. In a murine model of immunity to foreign RBCs, transfusion of mice with IgG-opsonized RBCs strongly attenuated the antibody response compared to transfusion of untreated RBCs. This model was used to study the anti-RBC IgG effect on the host immune response. Contrary to the predominant theories of the anti-D effect, here it is shown that IgG-mediated RBC clearance is not sufficient for the attenuation of antibody responses. IgG-opsonized RBCs internalized by the mononuclear phagocytic cells could stimulate T and B cell responses against RBC antigens. This thesis also shows that the adaptive tolerance at the T or B cell level is not the reason for the attenuation of the antibody response. Instead, IgG selectively prevented the appearance of antigen-primed RBC-specific B cells and, surprisingly, induced the host B cell response against the IgG in complex with RBCs. These results suggest that the inability of RBC-specific B cells to recognize and present RBC-specific epitopes may explain the inhibitory IgG effect.Ph