[Effects of an In-House Antibiotic Stewardship Initiative on Diagnosis and Management of Community-Acquired Pneumonia in Childhood and Adolescence]. / Ergebnisse der Einführung eines Antibiotic-Stewardship-Programms im stationären Bereich am Beispiel der ambulant erworbenen Pneumonie des Kindes- und Jugendalters.

BACKGROUND: Pediatric community-acquired pneumonia (pCAP) often leads to prescription of antibiotics and hospital admission of children. Unfortunately, adherence to diagnosis and treatment guidelines is inconsistent, and misuse of antibiotics may occur. Antimicrobial stewardship interventions, which were started in many hospitals during the last decade, can optimize management of pCAP without negative patient outcomes. OBJECTIVES: The objective of this retrospective analysis was to assess the influence of a newly implemented in-house pediatric antibiotic stewardship (ABS) initiative on guideline adherence and treatment quality. MATERIALS AND METHODS: Retrospective, patients' file-based analysis of the effects of an ABS initiative in a pediatric university hospital from January 2017 until March 2020. ABS initiative included creation of a local pCAP guideline for hospitalized children aged 90 days - 18 years, periodic training and continuous ABS support. RESULTS: A total of 230 patients with pCAP were included (145 before and 85 after intervention). Implementation of the ABS program led to reduction of antibiotics prescription without clear indication from 26 % to 10 % (p < 0.05). The inappropriate use of antibiotics decreased from 64 % to 27 % (p < 0.05), the rate of incorrect doses declined from 17 % to 10 % (p < 0.05) and the mean duration of antibiotic treatment declined from 10 to 7 days (p < 0.05). There were no differences between the two groups regarding length of stay, treatment failure or readmissions for respiratory infection. CONCLUSIONS: Pediatric antibiotic stewardship is an appropriate and safe method, and is beneficial to hospitalized patients with pCAP. Application of ABS programs may increase adherence to clinical guidelines and improve appropriate antimicrobial use without negative impact on patient outcomes. Multicenter follow-up studies are needed to clarify long-term effects of ABS programs.

Pathogenesis of varroosis at the level of the honey bee (Apis mellifera) colony.

The parasitic mite Varroa destructor, in interaction with different viruses, is the main cause of honey bee colony mortality in most parts of the world. Here we studied how effects of individual-level parasitization are reflected by the bee colony as a whole. We measured disease progression in an apiary of 24 hives with differing degree of mite infestation, and investigated its relationship to 28 biometrical, physiological and biochemical indicators. In early summer, when the most heavily infested colonies already showed reduced growth, an elevated ratio of brood to bees, as well as a strong presence of phenoloxidase/prophenoloxidase in hive bees were found to be predictors of the time of colony collapse. One month later, the learning performance of worker bees as well as the activity of glucose oxidase measured from head extracts were significantly linked to the timing of colony collapse. Colonies at the brink of collapse were characterized by reduced weight of winter bees and a strong increase in their relative body water content. Our data confirm the importance of the immune system, known from studies of individually-infested bees, for the pathogenesis of varroosis at colony level. However, they also show that single-bee effects cannot always be extrapolated to the colony as a whole. This fact, together with the prominent role of colony-level factors like the ratio between brood and bees for disease progression, stress the importance of the superorganismal dimension of Varroa research.

[Trauma of periorbital soft tissue. Reconstruction with regard to functional and aesthetic aspects]. / Trauma der periorbitalen Weichteile. Rekonstruktion unter funktionellen und ästhetischen Gesichtspunkten.

Complex midfacial trauma requires interdisciplinary management. A wide range of reconstructive procedures are needed and if necessary secondary and even tertiary interventions should be performed at the appropriate time. We present the case of a 45-year-old man who was involved in a car accident and presented with severe injuries to the periorbital soft tissue. The focus was on the reconstruction of this sensitive region with regard to functional and aesthetic aspects. In view of the severity of the injuries the final clinical status is considered to be very important. Special issues and pitfalls in the reconstruction of trauma cases and potential management are discussed.

Prophylactic intravenous nimodipine treatment in skull base surgery: pharmacokinetic aspects.

BACKGROUND: Nimodipine is primarily used in subarachnoid hemorrhage (SAH). Clinical trials revealed also a beneficial effect of prophylactic nimodipine treatment on cranial nerve functions following vestibular schwannoma surgery. OBJECTIVE: The unknown pharmacokinetics of prophylactically administered nimodipine were investigated. METHODS: Samples were taken from 27 patients with skull base lesions. Prophylactic intravenous nimodipine infusion was started 5.8-25.8 h (mean 17.9 h) before surgery. Nimodipine concentrations were determined in serum (intra- and postoperatively), cerebrospinal fluid (CSF) (intraoperatively), and tissue samples. RESULTS: Wide interindividual differences were observed. Mean concentrations for nimodipine were 46.9 ng/ml (SD: 6.4; min. 4.1 and max. 92.7 ng/ml) in intraoperative serum, 73.2 ng/ml (SD: 16.7; min. 6.6 and max. 253 ng/ml) in postoperative serum and 8.3 ng/ml (SD: 1.5; min. 1.0 und max. 29.7 ng/ml) in intraoperative CSF. The correlation between intra- and postoperative serum (p=0.004, r=0.560) and between intra-operative serum and CSF concentration (p=0.003, r=0.567) were statistically significant. Furthermore the correlation between intraoperative serum concentration and concentrations collected from vestibular nerves was high (r=0.711), but not statistically significant (p=0.178). CONCLUSIONS: Interindividually, continously administered intravenous nimodipine produces considerably variable serum levels. Controls of nimodipine serum concentrations may be useful to optimize nimodipine medication in skull base surgery and in the management of SAH. The serum nimodipine level is a useful marker for CSF and intracranial nerve tissue concentrations of nimodipine.

Nimodipine promotes regeneration of peripheral facial nerve function after traumatic injury following maxillofacial surgery: an off label pilot-study.

BACKGROUND: Animal tests, retro- and prospective clinical trials in neurosurgical departments have shown a beneficial effect of nimodipine on the preservation and recovery of facial and acoustic nerve function following vestibular schwannoma surgery. Encouraged by these positive results a pilot-study of nimodipine treatment in patients with a peripheral facial nerve (FN) paresis following maxillofacial surgery was performed. The rate and time of FN recovery were analysed and compared with the results in the literature. METHODS: Thirteen patients (n = 13) suffering from a moderate (1/13) up to a severe (12/13) peripheral FN paresis after maxillofacial surgery were treated with orally administered nimodipine. The anatomical main course of the FN was preserved in all patients with a 2nd to 3rd degree of Sunderland-injury (Sunderland, 1951). After no evidence of a spontaneous regeneration had shown, oral medication with nimodipine was started as an "off-label" use. RESULTS: An improvement of the FN function correlated to the start of the vasoactive medication and as a consequence a recovery of the FN function up to House-Brackmann (HB) grade I°-II° was observed in all the patients within a period of 2 months after the beginning of treatment (p = 0.00027). CONCLUSIONS: The clinical observations in these patients suggest a positive effect of nimodipine on the acceleration of peripheral FN regeneration after surgically caused trauma. The results of this pilot-study are very promising. A prospective study with a larger number of patients is planned to approve the beneficial effect of nimodipine on the peripheral FN in maxillofacial or otorhinolaryngological surgery.

In vitro investigation of the secondary palate development in two strains of mice.

The pathogenesis of cleft lip and palate (CL/P) is studied in animal experiments. This study revealed significant differences in foetal secondary palate development in two strains of mice (NMRI, A/WySnJ) using a palatal organ model. Palatal shelves of 114 NMRI embryos, resistant to cleft occurrence, and 93 A/WySnJ embryos, a strain with a high spontaneous CL/P rate, were micro-dissected at 14.25 GD (gestational day), before palatal fusion takes place. After cultivation in serum-free medium, palatal development was investigated microscopically and scored in a six-step system. At death (14.25 GD) the palatal shelves of the NMRI embryos (mean 3.5) were significant more developed than those of A/WySnJ (mean 2.7; p=0.05). After incubation, 53% (60/114) NMRI and 14% (13/93) A/WySnJ cultures had over two-thirds fusion to stage V-VI, therefore in 17% NMRI (19/114) and 1% A/WySnJ cultures (1/93) fusion was macroscopically complete. 62% of the A/WySnJ cultures showed no significant development in vitro (mean 2.84; p=0.094). There is a significant palatal development difference between normally developed NMRI (mean 4.45, p=0.05) and CL/P appearance in A/WySnJ mice (mean 2.84). Palatal development of both strains was significantly delayed in organ culture (p=0.05). The A/WySnJ strain was more susceptible to manipulation and vulnerable.

Pleiotropic effects of CXC chemokines in gastric carcinoma: differences in CXCL8 and CXCL1 expression between diffuse and intestinal types of gastric carcinoma.

CXC chemokines modulate host immunity, neovascularization, growth and invasive behaviour of tumours. Despite their relevance in tumour biology, chemokine expression in intestinal- and diffuse-type gastric carcinoma, which exhibit a completely different growth pattern, has not been investigated in detail. In this study, expression of the CXC chemokines CXCL8 [interleukin (IL)-8], CXCL1 [growth-related oncogene alpha (Gro alpha)], CXCL9 [monokine induced by interferon (IFN)-gamma] and CXCL10 [IFN-gamma-inducible protein-10 (IP-10)] and the corresponding chemokine receptors CXCR1-3 was investigated by immunohistochemistry in intestinal- and diffuse-type gastric carcinoma. Tumour cells of all patients expressed CXCL8. CXCL8 expression was significantly stronger in tumour cells of diffuse- rather than intestinal-type gastric carcinoma (P < 0.01) as determined by a semiquantitative score. CXCL1 was expressed almost exclusively by diffuse- but not intestinal-type carcinoma cells. The corresponding chemokine receptors, CXCR1 and CXCR2, were found on carcinoma cells. Furthermore, CXCL8 expression correlated with number of tumour vessels (P < 0.01), suggesting an angiogenetic function in gastric carcinoma not only in vitro but also in vivo. CXCL10 and CXCL9, attractants for T cells, were expressed by peritumorous macrophages in close proximity to IFN-gamma-producing CXCR3-positive T cells in both tumour types. These chemokines may attract gastric carcinoma-infiltrating T cells via an IFN-gamma-mediated pathway and enhance host immunity against the tumour. In gastric carcinoma a complex interplay between CXC-chemokine signals derived from both tumour cells and tumour-infiltrating immune cells may exhibit pleiotropic effects in tumour biology that go far beyond their originally described functions as leucocyte chemoattractants. Because CXCL8 and CXCL1, which are known to increase growth and invasive behaviour of malignant tumours, are significantly stronger expressed in diffuse- than intestinal-type gastric carcinoma, one may speculate that these chemokines influence the different growth pattern of gastric carcinoma types.

Functional dissection of the hexamerin receptor and its ligand arylphorin in the blowfly Calliphora vicina.

The process of receptor-mediated uptake of hexamerin storage proteins from insect haemolymph by fat body cells is a unique feature of the class Insecta. We identified the binding domains of the hexamerin receptor and the hexamerin ligand arylphorin in the blowfly, by means of the yeast-two-hybrid-system. The receptor-binding domain of arylphorin was located within domain 3 of the arylphorin monomer. The ligand-binding domain of the hexamerin receptor was mapped to the extreme N-terminus of the receptor. The binding domains identified exhibit no similarity to any functional protein domains known to date. Additionally, we identified two previously unknown protein-interactors of the hexamerin receptor. The results of this study provide further insights regarding the mechanism of the receptor-mediated endocytosis of storage proteins in insects.

Molecular cloning and localization of a cDNA encoding a crustacean hyperglycemic hormone from the South African spiny lobster, Jasus lalandii.

A cDNA, encoding a crustacean hyperglycemic hormone (cHH) of the South African spiny lobster, Jasus lalandii has been cloned. The cDNA consists of 1773 bp with an open reading frame of 399 bp that encodes a preprohormone of 133 amino acid residues. The preprohormone consists of a 25 amino acid hydrophobic signal peptide, a 32 amino acid cHH precursor-related peptide (CPRP) and the cHH sequence of 72 amino acid residues. The cHH sequence is flanked N-terminally by a Lys-Arg cleavage site and C-terminally by Gly-Lys, where Gly serves as an amidation site. The deduced amino acid sequence of the CPRP is in complete agreement with a peptide previously elucidated from sinus glands of J. lalandii, code-named CPRP 2 and the sequence of the cHH peptide matches that of the minor cHH isoform of J. lalandii, i.e. crustacean hyperglycemic hormone-II (cHH-II), which was also previously obtained by peptide sequencing. In situ hybridization on eyestalks revealed strong cHH-II mRNA expression in a subset of neurosecretory cells of the X-organ.

Complete sequence, expression and evolution of two members of the hexamerin protein family during the larval development of the rice moth, Corcyra cephalonica.

Three distinct types of storage hexamerins are expressed in the "last-instar" larvae of the rice moth, Corcyra cephalonica. A cDNA expression library was constructed from fat body-RNA and screened with a polyclonal antibody raised against purified hexamerin (SP2) of Corcyra cephalonica. Two slightly different "full-length" hexamerin cDNA clones (Hex2a and Hex2b) were isolated and sequenced. Both include open reading frames of 2109 bp which are translated into polypeptides of 703 amino acids with 92.5% identity. Signal peptides of 19 amino acids are present at the N-termini. The 684 amino acids native proteins have a high content of aryl groups (17.6%). According to both the criteria for amino acid composition and the phylogenetic analysis, Hex2a and Hex2b belong to the lepidopteran arylphorins. Northern blot studies revealed that the Hex2 genes are species- and tissue-specifically expressed in fat body cells of "last-instar" (= 5th) larvae.

Corazonin gene expression in the waxmoth Galleria mellonella.

We cloned and sequenced a full length cDNA coding for [Arg7]-corazonin in the greater wax moth Galleria mellonella. The deduced corazonin preprohormone consists of a nineteen amino acid signal peptide, the actual eleven amino acid corazonin sequence, followed by a Gly serving for amidation, a Lys-Arg processing site and an eighty amino acid corazonin precursor-related peptide. The data confirm the phylogenetic conservation of the actual corazonin sequence. The signal peptide and the precursor-related peptide exhibit a similar spacing of a few amino acids as detected in the corazonin preprohormone of Drosophila melanogaster. Northern blots and in situ hybridization experiments revealed that the G. mellonella corazonin gene is tissue-specifically expressed in four pairs of lateral neurosecretory cells in the brains of penultimate and last instar larvae, as well as of pupae and adults. No corazonin mRNA was detected in other cells of the nervous system, fat body, gut, and several other organs.

CXC chemokines Gro(alpha)/IL-8 and IP-10/MIG in Helicobacter pylori gastritis.

Infection with Helicobacter pylori causes chronic active gastritis, which is characterized by neutrophils infiltrating the gastric epithelial layer and the underlying lamina propria as well as by T, B lymphocytes and macrophages accumulating in the lamina propria. In this study, the chemokine profile responsible for the recruitment of these inflammatory cells is investigated. Using both RNA/RNA in situ hybridization and immunohistochemistry, the expression of the neutrophil and/or lymphocyte-attractant CXC chemokines growth-related oncogene alpha (Gro(alpha)), IL-8, interferon-gamma (IFN-gamma)-inducible protein-10 (IP-10), monokine induced by IFN-gamma (MIG) and the CC chemokines macrophage inflammatory protein-1alpha (MIP-1alpha), -1beta, regulated on activation normal T cell expressed and secreted (RANTES) and monocyte chemoattractant protein-1 (MCP-1) is studied and microanatomically localized in the gastric mucosa. Macrophages in the lamina propria at sites with neutrophil infiltration and gastric epithelium infiltrated by neutrophils highly expressed the neutrophil-attractant chemokines Gro(alpha) and IL-8. Additionally, Gro(alpha) and IL-8 were expressed by neutrophils themselves localized within gastric epithelium, in the foveolar lumen and in the cellular debris overlying mucosal erosion. IP-10 and to a lower extent MIG, both selectively chemotactic for inflammatory T cells, were expressed by endothelial cells of gastric mucosal vessels and by mononuclear cells at sites with T cell infiltration. Expression of all other CC chemokines tested was significantly lower. These in vivo data indicate that a set of predominantly CXC chemokines modulates the inflammation in H. pylori gastritis. Gro(alpha) and IL-8 may play an important role in neutrophil trafficking from the mucosal vessel into the gastric epithelium, whereas IP-10 and MIG contribute to the recruitment of inflammatory T cells into the mucosa.

Localization of glucocorticoid hormone receptors in mitochondria of human cells.

Glucocorticoid hormones regulate the transcription of nuclear genes by way of their cognate receptors. In addition, these hormones also modulate mitochondrial gene transcription by mechanisms which are as yet poorly understood. Using immunofluorescence labeling and confocal laser scanning microscopy we show that the glucocorticoid receptor of HeLa and Hep-2 cells is specifically enriched at the sites of the mitochondria which were visualized by labeling with the vital dye CMX and antibodies against cytochrome oxidase subunit I. Immunogold electron microscopy demonstrated that the receptor was located within the inner space of the mitochondria. Immunoblotting experiments also revealed the presence of glucocorticoid receptor in mitochondria isolated from HeLa and Hep-2 cells. Finally, living HeLa cells expressing green fluorescent-glucocorticoid receptor fusion protein revealed a distinct mitochondrial GFP fluorescence. Our results support the concept of a receptor-mediated direct action of steroid hormones on mitochondrial gene transcription.

Ligands and receptors: common theme in insect storage protein transport.

The passage of macromolecules through biological membranes is an essential process for all multicellular organisms. Insects have developed a mechanism different from that known for all other eukaryotes investigated so far. This review discusses the function and evolution of this mechanism. Insect pupae do not feed during metamorphosis. Therefore they depend on material that has been accumulated during the larval life. At the end of this period, shortly before pupariation, a rise in titer of ecdysteroid hormones induces the incorporation of a large fraction of storage proteins (hexamerins) from the body fluid into the fat body cells. The transport of hexamerins across the cell-membrane is mediated by a specific ecdysteroid-controlled receptor. It is synthesized as a precursor protein that is subsequently processed into the active receptor. This receptor protein is very unusual because it is closely related to its own hexamerin ligand. Sequence comparison shows that the hexamerins and hexamerin receptors diverged early in insect evolution and derive from a common hemocyanin ancestor.

Detection of oestrogen receptor variants in endometrium, myometrium, leiomyoma and peripheral blood mononuclear cells: comparison to variants present in breast cancer.

Oestradiol has mitogenic and regulatory effects on various organs and cells, mediated mainly by its nuclear receptor (ER). The presence of aberrant ER forms in Oestrogen-dependent tumours has been discussed in correlation with tumour progression. ER variants, generated by alternative splicing, have been detected in human breast cancer, but also in normal mammary glands, therefore their role in tumorigenesis has been questioned. We have investigated, by the use of the reverse transcription polymerase chain reaction amplification technique, the possible existence of ER variants in other normal oestrogen target organs and cells, such as uterus (myometrium and endometrium), in peripheral blood mononuclear cells and in a benign uterus tumour (leiomyoma). We have detected variant ER in these samples and have compared the variant profile to that observed in breast cancer. All tissues and cells studied expressed both wild-type ER and variant species. Variant forms encompassed ER with deletions of exons 2, 5 and 7. Variants with exon 5 deleted were detected only in peripheral blood mononuclear cells and in breast cancer. Variants with exons 2 and 7 deleted were present in all specimens tested. These results corroborate previous findings that the presence of ER variants is not a characteristic of breast cancer. The physiological significance and possible clinical relevance of the variant ER forms remain to be elucidated.

Complete sequence, expression, and evolution of the hexamerin LSP-2 of Calliphora vicina.

In cyclorraphan Diptera, two different types of hemolymph proteins exist which belong to the hexamerin family. During the last larval instar, Calliphora vicina synthesizes, besides the major fraction of arylphorin, a second hexameric protein, LSP-2. Here the developmentally regulated biosynthesis of this protein was analyzed. Western blot analyses showed that LSP-2 is not present in eggs, 1st, and 2nd instar larvae, whereas it can be detected in all tissues of last instar larvae. We report the characterization of the complete cDNA sequence that encodes a LSP-2 subunit, a nascent polypeptide of 701 amino acids with a molecular mass of 83.16 kDa. By Northern blotting, a mRNA of about 2.2 kb coding for LSP-2 is identified exclusively in the fat body of 3rd larval instars reflecting the stage and tissue specificity of LSP-2 gene expression. Phylogenetic analysis demonstrates the existence of two distinct groups of hexamerins in Diptera.

Characterization of the P25 silk gene and associated insertion elements in Galleria mellonella.

Insect silk genes attract attention by their precise territorial and developmental regulations and extremely high expression rates. Our present investigations demonstrated that the P25 silk gene of Galleria mellonella is down-regulated by ecdysteroid hormones. The gene was identified within 5217 nucleotides (nt) of two genomic clones. In contrast to other silk genes, Galleria P25 lacks the canonical TATA box. Transcription is initiated within a region of three nucleotides that lie at the end of a capsite initiator sequence ACAGT and about 90 nt downstream from a CAAT box. A stretch of 32 nt with a core sequence CTTTT was detected in the 5' region of Galleria P25 as well as in the presumptive regulatory regions of all other silk genes that are expressed in the posterior silk gland. However, consensus sequences reported for the regulatory regions of Bombyx silk genes are not obvious in Galleria P25. The coding sequence of this gene included 654 nt, is interrupted by 4 introns, and ends in position +3369; a potential polyadenylation signal starts at +4382. The gene contains 3 copies of a short interspersed nuclear element (SINE), which are located in the upstream region (-833 to -579) and in the first (+542 to +840) and second (+2259 to +2556) introns. The repeat, which was named Gm1, occurs in some other Galleria genes and exhibits homology to Bm1 SINE of the silkworm and to a similar element of a spider. Another insertion of at least 150 nt and with loosely defined borders is present in the 3' untranslated region (UTR) of Galleria P25. It includes a box (+3453 to +3552) of 99 nt that is tentatively called Lep1 because it was disclosed also in some other Lepidoptera. Lep1 seems to represent the core region of insertion elements that occur in the genomes of lepidopteran insects in various species specific and region specific modifications.