[Diabetes and Pregnancy]. / Diabetes mellitus und Schwangerschaft.

The quality of metabolic control at the beginning of pregnancy already determines the course and outcome of pregnanies with type 1 and type 2 diabetes mellitus. The preconceptional counseling and support provided by experienced teams is more important than modern technical equipment with insulin pumps and sensors for continuous glucose measurement. The incidence of congenital malformations is significantly reduced by a periconceptional HbA1c level < 6.5 % and folic acid supplementation started preconceptionally. To prevent preeclampsia, all women with type 1 and type 2 diabetes mellitus should be offered low-dose ASA, starting before 16 weeks of pregnancy. If the pregnant woman has a BMI < 25 kg/m² and persistently elevated fasting blood glucose levels, a GCK-MODY should be considered. For the diagnosis of asymptomatic gestational diabetes mellitus, all women in Germany with 24 + 0 to 27 + 6 weeks of pregnancy are offered a two-stage screening. Structured follow-up care is required after gestational diabetes mellitus, because these women have an increased risk of developing type 2 diabetes mellitus and cardiovascular complications. Pregnant women with COVID-19 and hyperglycemia have an increased risk of a severe course of the infection, which is further increased by obesity - they are an important target group for vaccination with an mRNA vaccine.

No Evidence for Ionotropic Pheromone Transduction in the Hawkmoth Manduca sexta.

Insect odorant receptors (ORs) are 7-transmembrane receptors with inverse membrane topology. They associate with the conserved ion channel Orco. As chaperon, Orco maintains ORs in cilia and, as pacemaker channel, Orco controls spontaneous activity in olfactory receptor neurons. Odorant binding to ORs opens OR-Orco receptor ion channel complexes in heterologous expression systems. It is unknown, whether this also occurs in vivo. As an alternative to this ionotropic transduction, experimental evidence is accumulating for metabotropic odor transduction, implicating that insect ORs couple to G-proteins. Resulting second messengers gate various ion channels. They generate the sensillum potential that elicits phasic-tonic action potentials (APs) followed by late, long-lasting pheromone responses. Because it is still unclear how and when Orco opens after odor-OR-binding, we used tip recordings to examine in vivo the effects of the Orco antagonist OLC15 and the amilorides MIA and HMA on bombykal transduction in the hawkmoth Manduca sexta. In contrast to OLC15 both amilorides decreased the pheromone-dependent sensillum potential amplitude and the frequency of the phasic AP response. Instead, OLC15 decreased spontaneous activity, increased latencies of phasic-, and decreased frequencies of late, long-lasting pheromone responses Zeitgebertime-dependently. Our results suggest no involvement for Orco in the primary transduction events, in contrast to amiloride-sensitive channels. Instead of an odor-gated ionotropic receptor, Orco rather acts as a voltage- and apparently second messenger-gated pacemaker channel controlling the membrane potential and hence threshold and kinetics of the pheromone response.

In situ tip-recordings found no evidence for an Orco-based ionotropic mechanism of pheromone-transduction in Manduca sexta.

The mechanisms of insect odor transduction are still controversial. Insect odorant receptors (ORs) are 7TM receptors with inverted membrane topology. They colocalize with a conserved coreceptor (Orco) with chaperone and ion channel function. Some studies suggest that insects employ exclusively ionotropic odor transduction via OR-Orco heteromers. Other studies provide evidence for different metabotropic odor transduction cascades, which employ second messenger-gated ion channel families for odor transduction. The hawkmoth Manduca sexta is an established model organism for studies of insect olfaction, also due to the availability of the hawkmoth-specific pheromone blend with its main component bombykal. Previous patch-clamp studies on primary cell cultures of M. sexta olfactory receptor neurons provided evidence for a pheromone-dependent activation of a phospholipase Cß. Pheromone application elicited a sequence of one rapid, apparently IP3-dependent, transient and two slower Ca(2+)-dependent inward currents. It remains unknown whether additionally an ionotropic pheromone-transduction mechanism is employed. If indeed an OR-Orco ion channel complex underlies an ionotropic mechanism, then Orco agonist-dependent opening of the OR-Orco channel pore should add up to pheromone-dependent opening of the pore. Here, in tip-recordings from intact pheromone-sensitive sensilla, perfusion with the Orco agonist VUAA1 did not increase pheromone-responses within the first 1000 ms. However, VUAA1 increased spontaneous activity of olfactory receptor neurons Zeitgebertime- and dose-dependently. We conclude that we find no evidence for an Orco-dependent ionotropic pheromone transduction cascade in M. sexta. Instead, in M. sexta Orco appears to be a slower, second messenger-dependent pacemaker channel which affects kinetics and threshold of pheromone-detection via changes of intracellular Ca(2+) baseline concentrations.

Transcriptional activity of the dominant gut mucosal microbiota in chronic inflammatory bowel disease patients.

Dysbiosis of the gut mucosa-associated microbiota (MAM) plays a pivotal role in the pathogenesis of chronic inflammatory bowel diseases (IBD). To date, dysbiosis only describes the altered composition of the different bacterial populations, but little is known about transcriptional activity, metabolism and the 'live' status of the MAM. In this study we investigated the transcriptional activity of the dominant intestinal bacterial populations in patients with IBD. Colonic mucosal biopsies from patients with active Crohn's disease (CD; n=10), active ulcerative colitis (UC; n=10) and healthy individuals (HI; n=10) were compared by 16S rRNA gene and rRNA profiles using clone libraries with more than 1700 sequenced clones. Bacterial richness was significantly lower in clone libraries based on rRNA compared to those based on the rRNA genes in the CD group (3.01 vs 3.91) and the UC group (3.61 vs 4.15), but showed no difference in HI (3.81 vs 3.85). The qualitative composition of rRNA and rRNA gene clone libraries was significantly different, with the phylum Bacteroidetes being the most active (P<0.01) compared to other populations in all clinical groups. In contrast, Actinobacteria and Firmicutes were inactive in the CD group, while Escherichia sp. were both abundant and active in the CD and UC groups. Most of the phylotypes showing the highest activity index ratios represented less than 1 % of the microbiota. Our findings indicate that specific bacterial populations are activated in IBD patients, while other groups are in an inactive or 'dormant' state. The transcriptional activity points to a more functional role of the intestinal mucosal microbiota and may lead to the identification of therapeutic targets in the active modulation of microbial factors.

The keyhole access expansion technique for flapless implant stage-two surgery: technical note.

In addition to osseointegration and restoration of function, patient satisfaction is a key element in the success of implant therapy. Especially in the esthetic zone, an essential part of the procedure aims at creating a definitive implant restoration that cannot be distinguished from the adjacent natural teeth. The present patient shows that, after localized ridge defects are reconstructed during implant surgery, a favorable esthetic and functional result can be achieved employing the keyhole access expansion technique for stage-two surgery, which is easy to perform, safe, and minimally invasive.

Perfusion with cAMP analogue affects pheromone-sensitive trichoid sensilla of the hawkmoth Manduca sexta in a time-dependent manner.

Octopamine causes time-dependent disadaptation of pheromone-sensitive olfactory receptor neurons (ORNs) of Manduca sexta. Because the majority of insect octopamine receptors are positively coupled to adenylyl cyclases we examined whether cyclic adenosine monophosphate (cAMP) mimics octopamine-dependent modulation of pheromone transduction in a time-dependent manner. Long-term tip recordings of single trichoid sensilla of Manduca sexta were performed during three zeitgeber times (ZTs, ZT 0=lights on), while stimulating the sensilla with two doses of the main pheromone component bombykal in a non-adapting protocol. The membrane-permeable cAMP analogue 8bcAMP increased the normalized sensillar potential amplitude in a time- and bombykal dose-dependent way. At the higher bombykal dose only, the applied 8bcAMP antagonized an endogenous decrease in the mean sensillar potential amplitude at ZT 1-4 and ZT 8-11 when ORNs were adapted but not at ZT 22-1, when ORNs were sensitized. In contrast to octopamine, 8bcAMP did not consistently affect the initial pheromone-dependent action potential frequency, the phasic/tonic response pattern, or the time-dependent shift to lower mean action potential frequencies at ZT 8-11. Furthermore, 8bcAMP increased the spontaneous action potential frequency time dependently, but differently from octopamine. In conclusion, our results show that cAMP only partly mimics the octopamine-dependent disadaptation of olfactory receptor neurons during photophase, apparently due to another missing octopamine-dependent synergistic factor such as defined intracellular calcium levels.

Multidimensional chromatography: a powerful tool for the analysis of membrane proteins in mouse brain.

Understanding the function of membrane proteins is of fundamental importance due to their crucial roles in many cellular processes and their direct association with human disorders. However, their analysis poses a special challenge, largely due to their highly amphipathic nature. Until recently, analyses of proteomic samples mainly were performed by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), due to the unprecedented separation power of the technique. However, in conventional 2D-PAGE membrane proteins are generally underrepresented due to their tendency to precipitate during isoelectric focusing and their inefficient transfer from the first to the second dimension. As a consequence, several other separation techniques, primarily based on liquid chromatography (LC), have been employed for analysis of this group of proteins. In the present study, different LC-based methods were compared for the analysis of crude protein extracts. One- and two-dimensional high-performance liquid chromatographic (1D- and 2D-HPLC) separations of brain protein tryptic digests with a predicted concentration range of up to 5 orders of magnitude were found to be insufficient, thus making a preceding fractionation step necessary. An additional protein separation step was introduced and a 3D-PAGE-HPLC analysis was performed. The results of these experiments are compared with results of 2D-PAGE/matrix-assisted laser desorption ionization mass spectrometric (MALDI MS) analyses of the same samples. Features, challenges, advantages, and disadvantages of the respective systems are discussed. The brain (mouse and human) was chosen as the analyzed tissue as it is of high interest in medical and pharmaceutical research into neurological diseases such as multiple sclerosis, stroke, Alzheimer's disease, and Parkinson's disease. The study is part of our ongoing research aimed at identifying new biomarkers for neurodegenerative diseases.

Pex19p-dependent targeting of Pex17p, a peripheral component of the peroxisomal protein import machinery.

Pex19p is required for the topogenesis of peroxisomal membrane proteins (PMPs). Here we have demonstrated that Pex19p is also required for the peroxisomal targeting and stability of Pex17p, a peripheral component of the docking complex of the peroxisomal protein import machinery. We have demonstrated that Pex17p is associated with the peroxisomal Pex13p-Pex14p complex as well as with Pex19p. We have identified the corresponding binding sites for Pex14p and Pex19p and demonstrated that a specific loss of the Pex19p interaction resulted in mistargeting of Pex17p. We have shown that a construct consisting only of the Pex19p- and Pex14p-binding sites of Pex17p is sufficient to direct an otherwise cytosolic reporter protein to the peroxisomal membrane in a Pex19p-dependent manner. Our data show that the function of Pex19p as chaperone or import receptor is not restricted to integral membrane proteins but may also include peripheral PMPs. As a consequence of our data, the previous definition of a targeting signal for PMPs (mPTS) as a Pex19p-binding motif in conjunction with a transmembrane segment should be extended to regions comprising a Pex19p-binding motif and a peroxisomal anchor sequence.