Genetic findings in miscarriages and their relation to the number of previous miscarriages.

PURPOSE: Early pregnancy loss leads to a devastating situation for many couples. Genetic disorders found in the pregnancy tissue are a frequent cause of miscarriages. It is unclear whether maternal age or previous miscarriages are associated with a higher chromosomal anomaly rate. This study aimed to determine the cytogenetical distribution of chromosomal disorders in couples after one or more previous miscarriages as well as the influence of maternal age. METHODS: 406 fetal tissue samples obtained after spontaneous abortion between 2010 and 2014 were successfully karyotyped. This included 132 couples with at least two losses and 274 couples with sporadic miscarriage. Normal and abnormal karyotype rate was determined for age, parity, gravidity, gestational week and number of previous miscarriages by logistic regression analysis. RESULTS: 145 (35.71%) fetal tissue samples had a normal karyotype, and 261 (64.8%) did not. After adjusting for age, older patients have a statistically significantly higher probability of genetic disorders in the pregnancy tissue (p < 0.001, OR 1.064, 95% CI 1.03-1.11). With each additional year, the probability of finding chromosomal abnormalities in a miscarriage increased by 6.4%. Patients younger than 35 years have a lower probability of having chromosomal disorders in the aborted material after two or more miscarriages than after sporadic miscarriages (50.7 vs. 58.9%) (p = 0.014, OR 0.67, 95% CI 0.48-0.914). Nevertheless, the risk of embryonic chromosomal disorders in patients aged 35 and above increased from 75.5% in sporadic miscarriages to 82.4% after more than one pregnancy losses (p = 0.59, OR 1.14, 95% CI - 0.72 to 1.92). CONCLUSION: Chromosomal disorders found after one or more previous miscarriages are related to patients' age. Couples suffering two or more miscarriages should be further researched, especially in younger patients.

Na(+) doping induced changes in the reduction and charge transport characteristics of Al2O3-stabilized, CuO-based materials for CO2 capture.

Chemical looping combustion (CLC) and chemical looping with oxygen uncoupling (CLOU) are emerging CO2 capture technologies that could reduce appreciably the costs associated with the capture of CO2. In CLC and CLOU, the oxygen required to combust a hydrocarbon is provided by a solid oxygen carrier. Among the transition metal oxides typically considered for CLC and CLOU, copper oxide (CuO) stands out owing to its high oxygen carrying capacity, exothermic reduction reactions and fast reduction kinetics. However, the low Tammann (sintering) temperature of CuO is a serious drawback. In this context, it has been proposed to support CuO on high Tammann temperature and low cost alumina (Al2O3), thus, reducing the morphological changes occurring over multiple CLC or CLOU redox cycles and stabilizing, in turn, the high activity of CuO. However, in CuO-Al2O3 systems, phase stabilization and avoiding the formation of the CuAl2O4 spinel is key to obtaining a material with a high redox stability and activity. Here, we report a Na(+) doping strategy to phase stabilize Al2O3-supported CuO, yielding in turn an inexpensive material with a high redox stability and CO2 capture efficiency. We also demonstrate that doping CuO-Al2O3 with Na(+) improves the oxygen uncoupling characteristics and coke resistance of the oxygen carriers. Utilizing in situ and ex situ X-ray absorption spectroscopy (XAS), the local structure of Cu and the reduction pathways of CuO were determined as a function of the Na(+) content and cycle number. Finally, using 4-point conductivity measurements, we confirm that doping of Al2O3-supported CuO with Na(+) lowers the activation energy for charge transport explaining conclusively the improved redox characteristics of the new oxygen carriers developed.

MID1, mutated in Opitz syndrome, encodes an ubiquitin ligase that targets phosphatase 2A for degradation.

The gene MID1, the mutation of which causes X-linked Opitz G/BBB syndrome (OS, MIM 300000), encodes a microtubule-associated protein (MAP). We show that mutation of MID1 leads to a marked accumulation of the catalytic subunit of protein phosphatase 2A (PP2Ac), a central cellular regulator. PP2Ac accumulation is caused by an impairment of a newly identified E3 ubiquitin ligase activity of the MID1 protein that normally targets PP2Ac for degradation through binding to its alpha4 regulatory subunit in an embryonic fibroblast line derived from a fetus with OS. Elevated PP2Ac causes hypophosphorylation of MAPs, a pathological mechanism that is consistent with the OS phenotype.

Opitz G/BBB syndrome, a defect of midline development, is due to mutations in a new RING finger gene on Xp22.

Opitz syndrome (OS) is an inherited disorder characterized by midline defects including hypertelorism, hypospadias, lip-palate-laryngotracheal clefts and imperforate anus. We have identified a new gene on Xp22, MID1 (Midline 1), which is disrupted in an OS patient carrying an X-chromosome inversion and is also mutated in several OS families. MID1 encodes a member of the B-box family of proteins, which contain protein-protein interaction domains, including a RING finger, and are implicated in fundamental processes such as body axis patterning and control of cell proliferation. The association of MID1 with OS suggests an important role for this gene in midline development.

Long-term follow-up of vascular reconstructions after supracondylar humerus fracture with vascular lesion in childhood.

INTRODUCTION: Supracondylar humerus fractures in childhood present with a pulseless but well-perfused hand in 2.6% of cases and with limb-threatening ischaemia in <1%. Conservative treatment is widely used in non-limb-threatening ischaemia, in particular if the child is very young (<2.5 years). It has been sufficiently proven that conservative treatment may retard growth. The aim of our study was to determine long-term patency rates after surgical reconstruction and growth impairment, if any, after surgical vascular reconstruction. PATIENTS AND METHODS: Between June 1990 and June 2004, 12 children (mean age 6.6 years, eight boys and four girls) with supracondylar fracture with vascular lesions underwent surgical reconstruction at the Department of Vascular Surgery at the University Hospital, Graz. Patient files were reviewed retrospectively. All patients were recalled for physical (forearm length and volume) and ultrasonographic examinations (forearm blood flow) in 2005 and for ultrasonographic examinations (reconstructed vascular area) in 2011, with a final mean follow-up time of 14.0 years (range 6.8-20.9 years). RESULTS: Twelve patients, 10 of whom had undergone growth measurements in 2005, were available for the latter examination. All were doing well, with patent vascular reconstructions. Seven reconstructed brachial arteries were enlarged, two of which with intramural calcifications, four did not show abnormalities and one presented with 45% thinning. There were no differences between affected and healthy forearms concerning volume, length and blood flow. CONCLUSIONS: Our data emphasise that surgical reconstruction is effective in terms of blood supply and growth. In cases with interposition of greater saphenous vein or venous patch plasty, we found a high risk for development of enlargements. We suggest that these patients be followed periodically, with ultrasound studies, to detect aneurysms and/or thrombotic changes as early as possible.

Dynamic longitudinal behavior in animals exposed to chronic social defeat stress.

Chronic social defeat (CSD) can lead to impairments in social interaction and other behaviors that are supposed to model features of major depressive disorder (MDD). Not all animals subjected to CSD, however, develop these impairments, and maintained social interaction in some animals is widely used as a model for resilience to stress-induced mental dysfunctions. So far, animals have mainly been studied shortly (24 hours and 7 days) after CSD exposure and longitudinal development of behavioral phenotypes in individual animals has been mostly neglected. We have analyzed social interaction and novel object recognition behavior of stressed mice at different time points after CSD and have found very dynamic courses of behavior of individual animals. Instead of the two groups, resilient or susceptible, that are found at early time points our data suggest four groups with (i, ii) animals behaving resilient or susceptible at early and late time points, respectively (iii) animals that start susceptible and recover with time or (iv) animals that are resilient at early time points but develop vulnerability later on.

[Dosage forms in the therapy of cardiovascular and circulatory diseases]. / Arzneiformen in der Herz-Kreislauftherapie.

A collection of dosage forms is presented, which are currently used in the treatment of cardiovascular and circulatory diseases including rarely used dosage forms. For each form important details to be considered during application are listed which are based on individual properties of the specific dosage form. Furthermore, general recommendations on the application of cardiovascular drugs are presented as well as information on dividing and crushing of dosage forms for dose reduction or application via tube, which is permitted for instant release dosage forms whereas modified release systems restrict the ability to divide or crush. Another chapter summarizes the prerequisites for a safe substitution of drugs which requires consideration of the particular dosage form.

Chaperone-assisted expression of authentic bovine adrenodoxin reductase in Escherichia coli.

Adrenodoxin reductase is an essential component of the mitochondrial monooxygenase systems that are involved in the synthesis of steroid hormones and related compounds. After removing by mutagenesis a secondary ribosome binding site and an mRNA loop formed between the gene and the vector, large amounts of the enzyme could be produced in Escherichia coli by coexpression with the HSP60-chaperone system. The purified protein was homogeneous enough for reproducible crystallization. The crystals diffracted X-rays isotropically beyond 1.7 A resolution permitting a structure analysis.

Molecular phylogeography of the nose-horned viper (Vipera ammodytes, Linnaeus (1758)): evidence for high genetic diversity and multiple refugia in the Balkan peninsula.

The nose-horned viper (Vipera ammodytes) occurs in a large part of the south-eastern Europe and Asia Minor. Phylogenetic relationships were reconstructed for a total of 59 specimens using sequences from three mitochondrial regions (16S and cytochrome b genes, and control region, totalling 2308 bp). A considerable number of clades were observed within this species, showing a large genetic diversity within the Balkan peninsula. Splitting of the basal clades was evaluated to about 4 million years ago. Genetic results are in contradiction with presently accepted taxonomy based on morphological characters: V. a. gregorwallneri and V. a. ruffoi do not display any genetic difference compared with the nominotypic subspecies (V. a. ammodytes), involving that these subspecies can be regarded as synonyms. High genetic divergence in the central part of the Balkan peninsula is not concordant with low morphological differentiation. Finally, the extensive genetic diversity within the Balkan peninsula and the colonisation routes are discussed.

[Intubation conditions and circulatory effects 90 seconds after a divided mivacurium dose with three different TIVA induction methods]. / Intubationsbedingungen und Kreislaufverhalten 90 s nach einer geteilten Mivacuriumdosis bei drei unterschiedlichen TIVA-Einleitungsformen.

UNLABELLED: The aim of this study was to compare the intubating conditions of a mivacurium-induced neuromuscular block 90 s after a divided administration with three different methods of induction of anaesthesia. METHODS: After approval by the local ethics committee, we investigated 36 ASA I and II patients undergoing a 2-h scheduled, elective surgery, in whom a TIVA was induced by one of three different drugs, edomidate, methohexital or propofol. After stable anaesthesia was reached, 0.15 mg/kg and 0.1 mg/kg of mivacurium, spaced 30 s apart, was injected. Endotracheal intubation was performed 90 s after the first micacurium injection and the intubation conditions were graded (1: excellent, 2: good, 3: poor; 4: impossible). The neuromuscular function was stimulated every 20 s by a nerve stimulator in a train-of-four (TOF) pattern, and the time to complete distinction of a TOF response as well as the time of reoccurrence of the first twitch was taken. A minute prior to injection of the relaxant and every minute for 5 min, the systolic and diastolic blood pressure, mean arterial pressure (MAP) and heart rate were measured. The neuromuscular block was maintained with a mivacurium infusion on a level of one twitch response. After cessation of the mivacurium infusion we recorded the time of reappearance of the second, third and fourth twitch responses. RESULTS: All patients could be intubated 90 s after mivacurium except for one, who was excluded for abnormal difficult intubation conditions. The etomidate group had significantly (chi 2 test) worse intubation grades than the methohexital group. In none of the groups did we observe any significant cardiovascular response due to the mivacurium injection, neither in blood pressure nor in heart rate. All groups showed similar onset of the maximal neuromuscular block (4 +/- 1.8 min) and recovery of the first TOF reaction (11.3 +/- 3.4 min). There was no difference in recovery from neuromuscular block maintained by infusion at the end of surgery. CONCLUSIONS: A dose of mivacurium 3.57 times the ED95 does not produce any haemodynamic instability, if it is divided into two parts to induce a TIVA. After this dose, all patients could be safely intubated within 90 s. A prolongation of the neuromuscular block after higher mivacurium doses could not be seen, and this dose did not produce a more rapid onset of the maximal block in any group. The time for recovery from a mivacurium infusion did not differ among the groups. Etomidate, due to its short half-life, seems not ideal for induction of a TIVA together with mivacurium in the dosage used. Mivacurium meets the demands of good controllability as required for a TIVA and can be recommended for a 90-s injection-intubation interval as well as for maintenance of the neuromuscular block.

Stable gene transfer and tissue-specific expression of a human globin gene using adenoviral vectors.

Helper-free double recombinant adenoviruses containing a genomic human globin gene and the neomycin resistance gene (neoR) have been constructed. The inserted globin and neoR genes are stable and transcription of two human globin genes (beta and a hybrid gamma-beta gene) is correctly initiated at the respective globin promoter during lytic infection in 293 cells. The neoR gene driven by the SV40 early promoter confers G418 resistance to human fibroblasts and K562 human erythro-leukemia cells transformed with these viruses. Most neoR clones contain the entire recombinant viral genome, including the inserted globin gene, integrated into their chromosomes. Normally, K562 cells express their gamma but not their beta globin genes. The transferred human beta globin gene was not expressed in either K562 cells or fibroblasts. However, the hybrid gamma-beta globin gene was expressed in all K562 clones that contained the gene whereas gamma-beta mRNA was barely detectable in the fibroblasts. This demonstrates tissue-specific expression of the adenovirus-transferred globin gene. Furthermore, the two transferred genes, globin and neoR, which are situated more than 20 kb apart in the viral genome appear to be independently regulated.

Retroviral-mediated transfer of genomic globin genes leads to regulated production of RNA and protein.

A high-titer amphotropic retroviral vector containing the neomycin resistance gene and a hybrid gamma-beta genomic human globin gene has been constructed. Mouse erythroleukemia cells infected with this virus were found to contain the full transcriptional unit of the transferred human globin gene by Southern blot analysis. These cells contain normally initiated, spliced, and terminated human globin mRNA. The human globin mRNA level increased 5- to 10-fold upon induction of the mouse erythroleukemia cells. Human globin chains were produced but only in a fraction of the cells as detected by immunofluorescent staining. A similar retrovirus containing a human beta-globin gene was used to transduce mouse erythroleukemia cells resulting in much higher levels of human globin synthesis than detected in mouse erythroleukemia cells transduced with the gamma-beta globin virus.

AIRE encodes a nuclear protein co-localizing with cytoskeletal filaments: altered sub-cellular distribution of mutants lacking the PHD zinc fingers.

The gene responsible for autoimmune polyendocrino-pathy candidiasis ectodermal dystrophy (APECED) recently has been positionally cloned to 21q22.3. This novel gene, AIRE, encodes for a predicted 57.7 kDa protein featuring two PHD-type zinc fingers shared by other proteins involved in chromatin-mediated tran-scriptional regulation. APECED is an autosomal recessive condition characterized by multiple polyendocrinopathies, and the typical triad of APECED symptoms includes hypoparathyroidism, primary adrenocortical failure and chronic mucocutaneous candidiasis. The aetiology of APECED is linked directly to mutations within the coding region of AIRE. These mutations are predicted to lead to truncated forms of the protein lacking at least one of the PHD zinc fingers. In this study, we have investigated the sub-cellular localization of AIRE expressed transiently in COS cells and fibroblasts. We found that AIRE has a dual nuclear and cytoplasmic localization. The wild-type protein is directed to speckled domains in the nucleus and also shows co-localization with cytoskeletal filaments. N-terminal AIRE fragments deleted for the PHD domain show altered nuclear localization, suggesting that the APECED mutations may elicit their primary effects in the nucleus.

Neuromuscular blockade by IgG antibodies from patients with Guillain-Barré syndrome: a macro-patch-clamp study.

Guillain-Barré syndrome (GBS) is often associated with serum antibodies to glycoconjugates such as GM1 and GQ1b. The pathogenic role of these antibodies and other serum factors has not yet been clarified. We have investigated the effect of serum, plasma filtrate, and highly purified IgG and IgM from 10 patients with typical GBS on motor nerve terminals in the mouse hemidiaphragm. Quantal endplate currents were recorded by means of a perfused macro-patch-clamp electrode. The plasma filtrate of all GBS patients led to a 5- to 20-fold reduction of evoked quantal release within 7 to 15 minutes of continuous superfusion. In 4 patients, the amplitudes of single quanta were clearly reduced (by 10-66% of control values), indicating an additional postsynaptic action. Blocking effects could be reversed to a variable degree within 15 to 18 minutes after washout. Purified IgG was as effective as native serum, whereas a purified GBS IgM fraction did not block transmission. Sera from convalescent patients and IgG from healthy subjects were without blocking effect. The effects were complement independent and there was no link to the presence (in 6 patients) or absence (in 4 patients) of detectable antibodies to GM1 or GQ1b. In GBS, antibodies to an undetermined antigen depress the presynaptic transmitter release and, in some cases, the activation of postsynaptic channels. We suggest that weakness in the acute stage of GBS may be caused in part by circulating antibodies.

DXS6673E encodes a predominantly nuclear protein, and its mouse ortholog DXHXS6673E is alternatively spliced in a developmental- and tissue-specific manner.

DXS6673E is a candidate gene for nonspecific X-linked mental retardation and encodes a novel Zn-finger protein. The ortholog murine gene DXHXS6673E in XC-D was isolated and characterized. It is ubiquitously expressed in all embryonic stages and adult tissues. Two different transcription start sites exist that result in two major transcripts of 6055 and 5352 nucleotides, each composed of 25 exons. Exon 1A is tissue specific, whereas exon 1B is transcribed constitutively. Both variants are translated into the same 1370-amino-acid protein. Transcripts are subject to alternative splicing at the 5'-end. Some of the isoforms are developmental stage and tissue specific. Among them, one was present only in embryos and adult brain. Sequence analysis demonstrated evolutionary conservation down to the arthropods and defined several conserved protein motifs. Subcellular localization studies with green fluorescent protein as a reporter showed that DXS6673E is predominantly located in the nucleus due to several functional nuclear localization signals. Three distinct protein distribution patterns in COS-7 cells could be identified.

The Opitz syndrome gene product, MID1, associates with microtubules.

Opitz syndrome (OS) is a genetically heterogeneous disorder characterized by defects of the ventral midline, including hypertelorism, cleft lip and palate, heart defects, and mental retardation. We recently identified the gene responsible for X-linked OS. The ubiquitously expressed gene product, MID1, is a member of the RING finger family. These proteins are characterized by an N-terminal tripartite protein-protein interaction domain and a conserved C terminus of unknown function. Unlike other RING finger proteins for which diverse cellular functions have been proposed, the function of MID1 is as yet undefined. By using the green fluorescent protein as a tag, we show here that MID1 is a microtubule-associated protein that influences microtubule dynamics in MID1-overexpressing cells. We confirm this observation by demonstrating a colocalization of MID1 and tubulin in subcellular fractions and the association of endogenous MID1 with microtubules after in vitro assembly. Furthermore, overexpressed MID1 proteins harboring mutations described in OS patients lack the capability to associate with microtubules, forming cytoplasmic clumps instead. These data give an idea of the possible molecular pathomechanism underlying the OS phenotype.