Reliability of growth prediction with hand-wrist radiographs

The aim of this study was to investigate the validity of hand-wrist radiographic analysis in estimating the amount of remaining craniofacial growth. The material compromised cephalograms of 22 males and 27 females with a Class I malocclusion. The median age of the females at the beginning (T1) was 11 years 10 months and of the males 12 years 6 months and at the end (T2) of treatment 14 years 7 months and 15 years 3 months, respectively. Statural height was measured and a lateral cephalogram was obtained for every patient at T1 and T2. A hand-wrist radiograph was taken only at T1. The cephalograms were scanned and analyzed. Relative dimensional growth changes in statural height as well as of the length of the cranial base (N-S), the maxilla (Ptm-A), and the dimensions of the mandible (Co-Gn, Go-Gn, and Co-Gn) from T1 to T2 were determined and statistically compared (Pearson's correlation coefficients) with the growth prediction assessed with the help of hand-wrist radiographs according to Greulich and Pyle. The results showed a highly significant correlation between statural growth increases and growth prediction assessed from the hand-wrist radiographs (females: r = 0.68; males: r = 0.7). Concerning craniofacial structures, the increase in mandibular corpus showed the highest correlation with growth prediction (females: r = 0.21; males: r = 0.52), but this association would not allow a reliable growth prediction. There was no significant correlation between growth increases of the cranial base, the maxilla, the ramus, and the effective length of the mandible and growth prediction assessed with the help of hand-wrist radiographs. As each patient has an individual growth pattern and different craniofacial structures show individual growth potential, it is questionable if quantitative craniofacial growth prediction with the help of hand-wrist radiographs is reliable. However, in an individual case for the assessment of the timing of the growth process, a hand-wrist radiograph can contribute to treatment plannin

Intestinal BMP-9 locally upregulates FGF19 and is down-regulated in obese patients with diabetes

believed to be mainly produced in the liver. The serum levels of BMP-9 were reported to be reduced in newly diagnosed diabetic patients and BMP-9 overexpression ameliorated steatosis in the high fat diet-induced obesity mouse model. Furthermore, injection of BMP-9 in mice enhanced expression of fibroblast growth factor (FGF)21. However, whether BMP-9 also regulates the expression of the related FGF19 is not clear. Because both FGF21 and 19 were described to protect the liver from steatosis, we have further investigated the role of BMP-9 in this context. We first analyzed BMP-9 levels in the serum of streptozotocin (STZ)-induced diabetic rats (a model of type I diabetes) and confirmed that BMP-9 serum levels decrease during diabetes. Microarray analyses of RNA samples from hepatic and intestinal tissue from BMP-9 KO- and wild-type mice (C57/Bl6 background) pointed to basal expression of BMP-9 in both organs and revealed a down-regulation of hepatic Fgf21 and intestinal Fgf19 in the KO mice. Next, we analyzed BMP-9 levels in a cohort of obese patients with or without diabetes. Serum BMP-9 levels did not correlate with diabetes, but hepatic BMP-9 mRNA expression negatively correlated with steatosis in those patients that did not yet develop diabetes. Likewise, hepatic BMP-9 expression also negatively correlated with serum LPS levels. In situ hybridization analyses confirmed intestinal BMP-9 expression. Intestinal (but not hepatic) BMP-9 mRNA levels were decreased with diabetes and positively correlated with intestinal E-Cadherin expression. In vitro studies using organoids demonstrated that BMP-9 directly induces FGF19 in gut but not hepatocyte organoids, whereas no evidence of a direct induction of hepatic FGF21 by BMP-9 was found. Consistent with the in vitro data, a correlation between intestinal BMP-9 and FGF19 mRNA expression was seen in the patients’ samples. In summary, our data confirm that BMP-9 is involved in diabetes development in humans and in the control of the FGF-axis. More importantly, our data imply that not only hepatic but also intestinal BMP-9 associates with diabetes and steatosis development and controls FGF19 expression. The data support the conclusion that increased levels of BMP-9 would most likely be beneficial under pre-steatotic conditions, making supplementation of BMP-9 an interesting new approach for future therapies aiming at prevention of the development of a metabolic syndrome and liver steatosis

Gene expression analysis of tumors demonstrates an induction of Th1 type immune response following intratumoral administration of ONCOS-102 in refractory solid tumor patients

2014-11-0

Methionine Sulfoxide Reductases Are Essential for Virulence of Salmonella Typhimurium

Production of reactive oxygen species represents a fundamental innate defense against microbes in a diversity of host organisms. Oxidative stress, amongst others, converts peptidyl and free methionine to a mixture of methionine-S- (Met-S-SO) and methionine-R-sulfoxides (Met-R-SO). To cope with such oxidative damage, methionine sulfoxide reductases MsrA and MsrB are known to reduce MetSOs, the former being specific for the S-form and the latter being specific for the R-form. However, at present the role of methionine sulfoxide reductases in the pathogenesis of intracellular bacterial pathogens has not been fully detailed. Here we show that deletion of msrA in the facultative intracellular pathogen Salmonella (S.) enterica serovar Typhimurium increased susceptibility to exogenous H2O2, and reduced bacterial replication inside activated macrophages, and in mice. In contrast, a ΔmsrB mutant showed the wild type phenotype. Recombinant MsrA was active against free and peptidyl Met-S-SO, whereas recombinant MsrB was only weakly active and specific for peptidyl Met-R-SO. This raised the question of whether an additional Met-R-SO reductase could play a role in the oxidative stress response of S. Typhimurium. MsrC is a methionine sulfoxide reductase previously shown to be specific for free Met-R-SO in Escherichia (E.) coli. We tested a ΔmsrC single mutant and a ΔmsrBΔmsrC double mutant under various stress conditions, and found that MsrC is essential for survival of S. Typhimurium following exposure to H2O2, as well as for growth in macrophages, and in mice. Hence, this study demonstrates that all three methionine sulfoxide reductases, MsrA, MsrB and MsrC, facilitate growth of a canonical intracellular pathogen during infection. Interestingly MsrC is specific for the repair of free methionine sulfoxide, pointing to an important role of this pathway in the oxidative stress response of Salmonella Typhimurium

Novel routes of hydroperoxide detoxification in Mycobacterium tuberculosis

Drug resistance and virulence of M. tuberculosis are in part related to the pathogen's antioxidant defense systems. KatG strains are resistant to the first line tuberculostatic isoniazid but they need to compensate their catalase deficiency by alternative peroxidase systems to remain virulent. So far only NADH-driven and AhpD-mediated hydroperoxide reduction by AhpC has been implicated as such a virulence-determining mechanism. In search of more likely metabolic links of AhpC, the genome of M. tuberculosis was screened for homologues of AhpF, thioredoxin-related proteins and peroxiredoxins. The genes were expressed heterologously in E. coli or M. smegmatis and purified to homogeneity. By means of those tools, two novel pathways were discovered that underscore the importance of the thioredoxin system for antioxidant defense in M. tuberculosis. They are: (1) NADPH-driven hydroperoxide reduction by AhpC that is mediated by thioredoxin reductase and thioredoxin C and (2) hydroperoxide reduction by the atypical peroxiredoxin TPx that equally depends on thioredoxin reductase but can use both thioredoxin B and C. According to kinetic analyses of hydroperoxide and peroxynitrite reduction and estimated enzyme levels, the redox cascade comprising thioredoxin reductase, thioredoxin C and TPx should be the most efficient system in situ to protect M. tuberculosis against oxidative and nitrosative stress.Antibiotika-Resistenz und Virulenz von Mycobacterium tuberculosis sind teilweise mit den antioxidativen Verteidigungssystemen des Pathogens verknüpft. KatG-Stämme sind Isoniazid-resistent. Sie müssen den KatG-Verlust durch alternative Peroxidase-Systeme ausgleichen, um ihre Virulenz erhalten zu können. Bisher ist nur ein NADH-gespeistes AhpD- vermitteltes Hydroperoxid-Reduktionssystem durch AhpC bekannt. Auf der Suche nach wahrscheinlicheren Reaktionspartnern der AhpC und weiteren Peroxidase-Systemen wurden in der vorliegenden Arbeit eine Vielzahl mykobakerieller Gene kloniert, heterolog exprimiert und durch funktionelle Analyse der isolierten Genprodukte auf Relevanz für den mykobakteriellen Peroxid-Stoffwechsel getestet. Die Auswahl der untersuchten Gene basierte auf bioinformatischen Homologie- bzw. Analogie-Analysen. In vitro Rekombination der Genprodukte ergab zwei neuartige Redoxkaskaden, die die Relevanz des Thioredoxin-Systems in Mykobakterien unterstreichen. Diese sind: 1. NADPH-abhängige Hydroperoxid-Reduktion durch AhpC, die durch TrxC und Thioredoxin-Reduktase vermittelt wird und 2. Hydroperoxid-Reduktion durch das atypische Peroxiredoxin TPx, die ebenfalls Thioredoxin-Reduktase- und TrxC- oder TrxB- vermittelt ist. Nach Maßgabe von in-vivo-Konzentrationen der beteiligten Enzyme und deren kinetischer Effizienz, ist der zweite beschriebene Weg der mykobakteriellen Peroxid- Detoxifikation in situ als maßgeblich einzustufen, um M. tuberculosis gegen oxidativen und nitrosativen Stress zu schützen

Molecular Dynamics Reveal Binding Mode of Glutathionylspermidine by Trypanothione Synthetase

The trypanothione synthetase (TryS) catalyses the two-step biosynthesis of trypanothione from spermidine and glutathione and is an attractive new drug target for the development of trypanocidal and antileishmanial drugs, especially since the structural information of TryS from Leishmania major has become available. Unfortunately, the TryS structure was solved without any of the substrates and lacks loop regions that are mechanistically important. This contribution describes docking and molecular dynamics simulations that led to further insights into trypanothione biosynthesis and, in particular, explains the binding modes of substrates for the second catalytic step. The structural model essentially confirm previously proposed binding sites for glutathione, ATP and two $Mg^{2+}$ ions, which appear identical for both catalytic steps. The analysis of an unsolved loop region near the proposed spermidine binding site revealed a new pocket that was demonstrated to bind glutathionylspermidine in an inverted orientation. For the second step of trypanothione synthesis glutathionylspermidine is bound in a way that preferentially allows $N^1$-glutathionylation of $N^8$-glutathionylspermidine, classifying $N^8$-glutathionylspermidine as the favoured substrate. By inhibitor docking, the binding site for $N^8$-glutathionylspermidine was characterised as druggable