Molecular surveillance of carbapenem-resistant Enterobacterales in two nearby tertiary hospitals to identify regional spread of high-risk clones in Germany, 2019-2020.

BACKGROUND: Understanding the transmission dynamics of carbapenem-resistant Enterobacterales (CRE) is critical to addressing the escalating global threat of antimicrobial resistance (AMR). Although hospital transmission of CRE has been extensively studied, information on community transmission is lacking. AIM: To identify genomic clusters of CRE from two nearby institutions that may be indicative of community or inter-facility transmission. METHODS: CRE isolates between January 1st, 2019 and December 31st, 2020 from two tertiary hospitals, detected in the respective routine microbiology laboratories, were collected and characterized by short-read whole-genome sequencing. FINDINGS: A total of 272 CRE were collected, with Enterobacter cloacae complex (71/192, 37%) predominant in Heidelberg and Escherichia coli (19/80, 24%) in Mannheim. The most common carbapenem resistance gene, blaOXA-48, was detected in 38% of CRE from both centres. Several putative transmission clusters were found, including six clusters of E. cloacae complex, five clusters of Klebsiella pneumoniae, four clusters of Citrobacter freundii, and two clusters each of Escherichia coli and K. aerogenes. No clusters involved isolates from both study centres, except for an ST22 C. freundii cluster. Globally circulating clones were identified between the two centres for ST131 E. coli, ST66 E. hormaechei, and ST22 C. freundii. CONCLUSION: This study found no widespread transmission clusters among isolates from both centres, suggesting a hospital-specific clonal structure. This suggests that CRE clusters involving both institutions may indicate emerging or circulating clones in the community, highlighting the need for intersectoral surveillance and data sharing.

Awareness of Age-Related Macular Degeneration in Community-Dwelling Elderly Persons in Germany.

Background: Due to current demographic trends age-related macular degeneration (AMD) is becoming more prevalent. When disease progresses to late-stage neovascular AMD, rapid initiation of treatment is required to achieve optimal outcomes. However, many affected individuals may be unaware of their disease impeding and delaying care seeking. Therefore, in an exploratory study we assessed whether elderly persons living independently in the community were aware of their AMD. Methods: Participants were recruited in eleven seniors' community centers. Participants underwent a standardized interview followed by non-mydriatic fundus photography of the macula and the optic disc in both eyes (Canon CR-2AF, Canon, New York, USA). The images were graded by an ophthalmologist and the data were analyzed descriptively. Results: A total of 281 participants (73.9 ± 8.1 years; 71.9% women) underwent bilateral fundus photography. The fundus photographs of 208 participants (74%; 73.6 ± 7.0 years; 73.1% women) could be graded. In a third (32.2%, n = 67) no pathological changes were detected. AMD was present in 24.5% of the examined subjects (n = 51). Half of the cases had early (47.1%), followed by intermediate (41.2%) and late (11.7%) AMD. Only one third (n = 16, 31.4%) were aware of their disease. Conclusions: A quarter of community dwelling elderly had AMD but only a third of these were aware of being affected with AMD. This confirms previous studies demonstrating low awareness for age-related eye diseases in the community. Considering the increase in population aging, awareness campaigns for AMD are needed.

[Physical activity in older persons with eye diseases : Applicability of wrist-worn accelerometer]. / Physische Aktivität bei älteren Menschen mit Augenerkrankungen : Anwendbarkeit von Armbandakzelerometern.

BACKGROUND AND OBJECTIVES: Physical activity (PA) impacts age-related diseases but its measurement is difficult. The acceptance of wrist-worn accelerometers (ACC) in older patients with eye diseases was evaluated and the results were compared with a validated activity questionnaire. MATERIALS AND METHODS: In this study 50 patients underwent a clinical examination and were interviewed with the International Physical Activity Questionnaire (IPAQ). They then wore an ACC for 7 days and then for 30 days. After descriptive analysis of the data, influencing factors on the PA were assessed using multiple, linear models. RESULTS: A total of 94% of participants wore the ACC for 7 days and 74% for 30 days. For 36 patients complete data were available. In comparison, IPAQ (e. g. moderate PA: 1183.4 ± 864.3 min/week) vs. ACC across 7 and 30 days (248.5 ± 266.0 min/week and 248.8 ± 190.6 min/week, respectively) showed substantially higher results (p < 0.05 for both intervals). The ACC data across 7 and 30 days were comparable (e. g. high PA: 25.4 ± 33.7 min/week and 22.5 ± 29.0 min/week). In multiple, linear models body mass index (BMI) was associated with 7 days moderate activity (ß = -0.22 [95% confidence intervals CI: -21.7; -1.9]; p = 0.021) and vigorous activity (ß = -0.35 [95% CI: -4.3; -0.5]; p = 0.033) as well as 30 days vigorous activity (ß = -0.45 [95% CI: -5.9; -0.1]; p = 0.044). Other factors such as better visual acuity, age and gender were not associated. CONCLUSION: The use of ACCs are acceptable to the majority of older patients with eye diseases. The results of ACC are more precise with lower variation, and much lower PA than those of a comparable activity questionnaire. Measurement over 7 days is sufficient for capturing the average PA in older patients and can be easily applied in clinical trials.

[Agitation in prehospital setting: view of emergency physicians]. / Prise en charge des états d'agitation extrahospitaliers: le point de vue de l'urgentiste.

Agitation is a behavioral emergency that can have numerous underlying causes, organic as well as psychiatric. The presence of a clinician and well-trained staff determine the quality of care, thus facilitating the dialog with the patient and, if needed, ensuring the safe application of medication and physical restraint. Outside the hospital, an emergency physician is best suited to manage such situations in collaboration with paramedics and police. "De-escalation" without medication or physical restraint should be the preferred end-point of an intervention in agitation. When necessary, medication and restraint should be applied, in order to enable medical evaluation of a patient. Calming without sedation seems the most advised endpoint of intervention, according to expert-opinion.

Absence of APOBEC-1 mediated mRNA editing in human carcinomas.

The transgene expression of the catalytic subunit APOBEC-1 of the apo B mRNA editing enzyme-complex can cause hepatocellular carcinoma in mice and rabbits. It has been proposed that aberrant editing of mRNA may represent a novel oncogenic principle. This investigation aimed to define whether such aberrant hyperediting mediated by APOBEC-1 occurs in human carcinomas. Editing and hyperediting of apo B, NAT1 or NF1 mRNA was not identified in any of 28 resected tumor specimens, including hepatocellular, bile duct, gastric, colorectal, pancreatic adeno- and neuroendocrine, lung adeno-, medullary thyroid and breast carcinoma, soft tissue sarcoma and neuroblastoma. In most types of carcinoma, significant levels for full-length APOBEC-1 mRNA could not be detected. Low level expression of APOBEC-1 was found in colorectal and gastric carcinoma where most of the APOBEC-1 mRNA is inactivated by alternate splicing. The 'auxiliary' components of the apo B mRNA editing enzyme-complex are missing in many tumors including colorectal and gastric carcinoma, but are highly expressed in hepatocellular, lung adeno- and breast carcinoma all of which lack APOBEC-1. Taken together, either APOBEC-1 or the 'auxiliary' components of the apo B mRNA editing enzyme-complex or both are missing in human carcinomas resulting in the absence of mRNA editing. Currently, there is no evidence that aberrant editing mediated by APOBEC-1 contributes to the tumorigenesis of natural human carcinomas.

Interference of H2O2 with stimulus-secretion coupling in mouse pancreatic beta-cells.

1. We have reported previously that in mouse pancreatic beta-cells H2O2 hyperpolarizes the membrane and increases the ATP-sensitive K+ current recorded in the perforated patch configuration of the patch-clamp technique. The present study was undertaken to elucidate the underlying mechanisms. 2. The intracellular ATP concentration measured by chemoluminescence was reduced by H2O2. The ADP concentration increased in parallel during the first 10 min, resulting in a pronounced decrease in the ATP/ADP ratio. 3. Consistent with these results, glucose-stimulated insulin secretion from isolated islets was inhibited by H2O2. 4. Membrane hyperpolarization measured with intracellular microelectrodes in intact islets and inhibition of insulin secretion were counteracted by tolbutamide, indicating that the channels are still responsive to inhibitors and that the ATP concentration is not too low to trigger exocytosis. However, the sensitivity of the beta-cells to tolbutamide was reduced after treatment with H2O2. 5. H2O2 increased the intracellular Ca2+ activity ([Ca2+]i) in a biphasic manner. A first transient rise in [Ca2+]i due to mobilization of Ca2+ from intracellular stores was followed by a sustained increase, which was at least partly dependent on Ca2+ influx. The first phase seems to reflect Ca2+ mobilization from mitochondria. 6. Our results demonstrate that H2O2 interferes with glucose metabolism, which influences the membrane potential and ATP-sensitive K+ current via the intracellular concentration of ATP. These events finally lead to an inhibition of insulin secretion despite an increase in [Ca2+]i.

Distinct promoters induce APOBEC-1 expression in rat liver and intestine.

The expression of apolipoprotein (apo) B can be modulated by mRNA editing, a unique posttranscriptional base change in the apo B mRNA. Apo B-48, the translation product of edited apo B mRNA, is not a precursor of the atherogenic low density lipoproteins and lipoprotein(a). In humans and various other mammals, the apo B mRNA is edited in the intestine but not in the liver, which exclusively secretes apo B-100-containing lipoproteins as precursors for low density lipoprotein formation. In species such as the rat, mouse, dog, and horse, apo B mRNA is also edited in the liver, resulting in low plasma levels of low density lipoprotein. Editing of the apo B mRNA is mediated by the apo B mRNA-editing enzyme complex, of which the catalytic subunit APOBEC-1 is not expressed in the liver of species without hepatic editing. To understand the molecular basis for liver-specific expression of APOBEC-1 and the editing of hepatic apo B mRNA, the expression pattern and genomic organization of the rat APOBEC-1 gene have been characterized. The rat APOBEC-1 gene contains 6 exons and 2 promoters with distinct activities. The expression of APOBEC-1 in the rat liver is the result of a promoter located upstream, with tissue-specific exon use and alternate splicing within the 5'-untranslated region of APOBEC-1 mRNA encoded by exon 2. In addition to the liver, this promoter also induces APOBEC-1 expression in the spleen, lung, kidney, heart, and skeletal muscle. The promoter located downstream belongs to a new class of TATA-less promoters and is responsible for the abundant expression of APOBEC-1 in the intestine. Mapping of the transcriptional start sites and deletion analysis of the promoter regions by using luciferase as the reporter gene have defined the regulatory elements of both promoters. The downstream, intestine-specific promoter contains a negative regulatory element between -1100 and -500, which appears to restrict its activity to the intestine. The upstream, liver-specific promoter of the rat APOBEC-1 gene induces APOBEC-1 expression and editing of apo B mRNA in human hepatoma HuH-7 and Hep G2 cells. Understanding the molecular basis for the liver-specific expression of APOBEC-1 in the rat promises new strategies to induce APOBEC-1 expression in the human liver for the reduction of atherogenic lipoprotein levels by hepatic apo B mRNA editing.

Imidazole antimycotics affect the activity of various ion channels and insulin secretion in mouse pancreatic B-cells.

In the present paper the effects of antimycotics with imidazole structure on the activity of various ion currents of mouse pancreatic B-cells and insulin secretion from isolated islets have been studied. Clotrimazole (0.1-10 microM, bath solution without albumin) reversibly inhibited the whole-cell K + ATP current studied with the patch-clamp technique and concomitantly depolarized the membrane potential. Two other structurally related compounds, econazole and ketoconazole, exhibited similar effects on the whole-cell K + ATP current. Clotrimazole also inhibited the current through single K + ATP channels measured in the inside-out configuration. According to these results it seems unlikely that a cytoplasmic factor is involved in the action of clotrimazole on K + ATP currents. Clotrimazole (10 microM) also reduced the current through voltage-dependent Ca2+ and K+ channels and altered inactivation kinetics. Moreover, clotrimazole reversibly abolished a recently described inward current which is induced by hypotonic cell swelling. The results show that clotrimazole altered the activity of all ion currents in B-cells investigated in this study. Clotrimazole (3-100 microM, solution with albumin) irreversibly inhibited insulin secretion from isolated islets. With econazole and ketoconazole similar effects on hormone release were observed. The changes in the activity and kinetics of voltage-dependent Ca2+ and K+ currents are likely to contribute to the observed inhibition of insulin secretion. However, we cannot entirely rule out that imidazole antimycotics also interfere with a step in stimulus-secretion coupling distal to changes in membrane potential.

Effects of the nitric oxide synthase inhibitor N omega nitro-L-arginine methyl ester on electrical activity and ion channels of mouse pancreatic B cells.

The effects of the nitric oxide (NO) synthase inhibitor N omega-nitro-L-arginine methyl ester (L-NAME) were studied on electrical activity, K+ATP currents and voltage-dependent K+ and Ca2+ channel currents in mouse pancreatic B cells. In the presence of 15 mM glucose, L-NAME (> 5 mM) depolarized the B cell membrane and electrical activity became continuous. The depolarization caused by L-NAME was not reversed by the addition of a membrane permeant cyclic GMP analogue. L-NAME inhibited the whole-cell K+ATP current reversibly by reducing the single channel current conductance and open probability. The results are consistent with the assumption that L-NAME depolarizes the B cell membrane by a direct action on the K+ATP channel. L-NAME also influenced the whole-cell current through voltage-dependent K+ and Ca2+ channels.

The effects of nitric oxide on the membrane potential and ionic currents of mouse pancreatic B cells.

Nitric oxide (NO) is considered to contribute to the impairment of B cell function in insulin-dependent diabetes mellitus. The effects of compounds that release NO were tested on the membrane potential and ionic currents of mouse pancreatic B cells using intracellular microelectrodes and the whole-cell patch-clamp technique. S-Nitrosocysteine led to a concentration-dependent reduction of electrical activity induced by 15 mM glucose. At a concentration of 1 mM, S-nitrosocysteine cause a hyperpolarization of the plasma membrane with complete suppression of electrical activity. In about half of the cells tested, electrical activity reappeared during treatment with S-nitroso-cysteine or after wash-out. However, in the other cells the hyperpolarization was followed by a slow depolarization and electrical activity did not reappear. The perforated-patch whole-cell K+ATP current first increased and subsequently decreased again during exposure to 1 mM S-nitroso-cysteine. With 0.1 and 0.01 mM S-nitroso-cysteine, only the rise of the current amplitude was observed. S-nitroso-cysteine (1 mM) almost completely abolished the current through voltage-dependent Ca2+ channels (measured with Ba2+ as charge carrier). Like S-nitroso-cysteine, 100 microM sodium-nitroprusside, another donor, evoked a marked hyperpolarization of the membrane potential that was at least in part reversible. To further ascertain that the effect of S-nitroso-cysteine was mediated by NO, we tested the decomposition products of S-nitroso-cysteine. Nitrite and denitrosylated S-nitroso-cystein (1 mM) did not alter electrical activity of B cells, whereas cysteine (1 mM) caused a slight depolarization. It is concluded that exogenous NO evokes rapid changes of B cell function by influencing the activity of ion channels.