Insulin and glycolysis dependency of cardioprotection by nicotinamide riboside.

Decreased nicotinamide adenine dinucleotide (NAD+) levels contribute to various pathologies such as ageing, diabetes, heart failure and ischemia-reperfusion injury (IRI). Nicotinamide riboside (NR) has emerged as a promising therapeutic NAD+ precursor due to efficient NAD+ elevation and was recently shown to be the only agent able to reduce cardiac IRI in models employing clinically relevant anesthesia. However, through which metabolic pathway(s) NR mediates IRI protection remains unknown. Furthermore, the influence of insulin, a known modulator of cardioprotective efficacy, on the protective effects of NR has not been investigated. Here, we used the isolated mouse heart allowing cardiac metabolic control to investigate: (1) whether NR can protect the isolated heart against IRI, (2) the metabolic pathways underlying NR-mediated protection, and (3) whether insulin abrogates NR protection. NR protection against cardiac IRI and effects on metabolic pathways employing metabolomics for determination of changes in metabolic intermediates, and 13C-glucose fluxomics for determination of metabolic pathway activities (glycolysis, pentose phosphate pathway (PPP) and mitochondrial/tricarboxylic acid cycle (TCA cycle) activities), were examined in isolated C57BL/6N mouse hearts perfused with either (a) glucose + fatty acids (FA) ("mild glycolysis group"), (b) lactate + pyruvate + FA ("no glycolysis group"), or (c) glucose + FA + insulin ("high glycolysis group"). NR increased cardiac NAD+ in all three metabolic groups. In glucose + FA perfused hearts, NR reduced IR injury, increased glycolytic intermediate phosphoenolpyruvate (PEP), TCA intermediate succinate and PPP intermediates ribose-5P (R5P) / sedoheptulose-7P (S7P), and was associated with activated glycolysis, without changes in TCA cycle or PPP activities. In the "no glycolysis" hearts, NR protection was lost, whereas NR still increased S7P. In the insulin hearts, glycolysis was largely accelerated, and NR protection abrogated. NR still increased PPP intermediates, with now high 13C-labeling of S7P, but NR was unable to increase metabolic pathway activities, including glycolysis. Protection by NR against IRI is only present in hearts with low glycolysis, and is associated with activation of glycolysis. When activation of glycolysis was prevented, through either examining "no glycolysis" hearts or "high glycolysis" hearts, NR protection was abolished. The data suggest that NR's acute cardioprotective effects are mediated through glycolysis activation and are lost in the presence of insulin because of already elevated glycolysis.

Ferroelastic Twin Angles at the Surface of CaTiO_{3} Quantified by Photoemission Electron Microscopy.

We use photoemission electron microscopy to measure the ferroelastic twin wall angles at the surface of CaTiO_{3} (001) and deduce the strain ordering. We analyze the angular dependence of the photoelectron emission from different domain surfaces, each with its own characteristic tilt angle in the factory rooflike topography. By considering the surface topography as a field perturbation, the offset in the photoemission threshold can be directly related to the tilt angles. With knowledge of the symmetry allowed twin walls we quantify the twin angles between 179.1° to 180.8°.

Multi-criteria analysis of strategies towards sustainable recycling of phosphorus from sewage sludge in Austria.

To promote optimal phosphorus (P) recovery from municipal wastewater and sewage sludge with viable legal instruments, it is imperative to understand the regional and national consequences of different legal requirements for recycling. In this study we develop a scenario-based analysis to assess the environmental and economic impact of different national P recovery strategies in the context of a detailed representation of the existing Austrian wastewater infrastructure. This assessment combines material flow analysis, life cycle assessment and life cycle costing and includes the indicators P recycling rate, P utilization degree, heavy metal removal rate, share of heavy metals' content in wastewater redirected to agricultural soils, global warming potential, cumulated energy demand, terrestrial acidification potential, volume of freight transport and annual costs. The following main conclusions can be drawn. P recovery from ash shows the highest potential regarding the utilization of P from wastewater. A high P utilization from wastewater should rely on recovery technologies that decontaminate products, otherwise pollutant loads to agricultural soils might increase. P recovery to the extent of 60-85 % of P in WWTPs influent can be achieved by savings/costs of -0.8 to +4.7 EUR inhabitant-1 yr-1 in addition to current cost of the wastewater treatment/sludge disposal system. Key factors to be considered for costs are the choice of recovery process, revenues from products, and the use of existing incineration infrastructure. P recovery can lead to the reduction of greenhouse gas emissions in Austria if nitrous oxide emissions from sludge incineration are limited and efficient heat utilization strategies are implemented. There is a trade-off in terms of environmental and economic costs in choosing a more centralized or decentralized mono-incineration strategy.

Two phase-matching regimes in high-order harmonic generation.

High-order harmonic generation (HHG) provides scalable sources of coherent extreme ultraviolet radiation with pulse duration down to the attosecond time scale. Efficient HHG requires the constructive interplay between microscopic and macroscopic effects in the generation volume, which can be achieved over a large range of experimental parameters from the driving field properties to those of the generating medium. Here, we present a systematic study of the harmonic yield as a function of gas pressure and medium length. Two regimes for optimum yield are identified, supporting the predictions of a recently proposed analytical model. Our observations are independent on the focusing geometry and, to a large extent, on the pulse duration and laser intensity, providing a versatile approach to HHG optimization.

Chemical and mechanical activation of resident cardiac macrophages in the living myocardial slice ex vivo model.

Resident cardiac macrophages (rcMACs) are among the most abundant immune cells in the heart. Plasticity and activation are hallmarks of rcMACs in response to changes in the microenvironment, which is essential for in vitro experimentation. The in vivo investigation is confounded by the infiltration of other cells hindering direct studies of rcMACs. As a tool to investigate rcMACs, we applied the ex vivo model of living myocardial slices (LMS). LMS are ultrathin ex vivo multicellular cardiac preparations in which the circulatory network is interrupted. The absence of infiltration in this model enables the investigation of the rcMACs response to immunomodulatory and mechanical stimulations. Such conditions were generated by applying interferon-gamma (IFN-γ) or interleukine-4 (IL-4) and altering the preload of cultured LMS, respectively. The immunomodulatory stimulation of the LMS induced alterations of the gene expression pattern without affecting tissue contractility. Following 24 h culture, low input RNA sequencing of rcMACs isolated from LMS was used for gene ontology analysis. Reducing the tissue stretch (unloading) of LMS altered the gene ontology clusters of isolated rcMACs with intermediate semantic similarity to IFN-γ triggered reaction. Through the overlap of genes affected by IFN-γ and unloading, we identified Allograft inflammatory factor 1 (AIF-1) as a potential marker gene for inflammation of rcMACs as significantly altered in whole immunomodulated LMS. MicroRNAs associated with the transcriptomic changes of rcMACs in unloaded LMS were identified in silico. Here, we demonstrate the approach of LMS to understand load-triggered cardiac inflammation and, thus, identify potential translationally important therapeutic targets.

[Prevalence of hypophosphatasia in adult patients in rheumatology]. / Prävalenz der Hypophosphatasie bei adulten Patienten in der Rheumatologie.

BACKGROUND: Hypophosphatasia (HPP) is a genetic disorder caused by one or more mutations in the alkaline phosphatase (ALP) gene, responsible for encoding tissue-specific ALP and for the mineralization process. OBJECTIVE: Identification of the prevalence of HPP in rheumatology patients. MATERIAL AND METHODS: Medical records of all adult rheumatology patients with pathologically low total ALP levels (<35 U/L) treated in the Department of Rheumatology at the Clinic of Internal Medicine III, University Hospital Bonn between January 2017 and June 2019, were retrospectively examined for clinical signs as well as for results of genetic tests for HPP. RESULTS: In 60 out of 2289 patients (2.62%) pathologically low ALP levels were detected. Of these 30 (1.31%) were found to have persistently low ALP levels. Genetic testing for ALP gene mutations was performed in 19 of these 30 patients and 7 of the 19 patients (36.84%) had HPP signs (insufficiency fractures, or bad dental status since childhood), all with pathologic ALP mutations. Of these patients 3 (15.78%) each had a history of insufficiency fracture with normal bone densitometry. Overall, 13 out of the 19 patients (68.42%) had mutations in the ALP gene. Interestingly, no association with chondrocalcinosis was detected in any of the patients. CONCLUSION: The HPP seems to be an underdiagnosed disease with a higher proportion of affected rheumatology patients. Therefore, future studies should aim to develop a diagnostic protocol in the clinical practice.

Long-term effects of splint therapy in patients with posttraumatic stress disease (PTSD).

OBJECTIVES: The aim of a pilot study was to clarify the question of whether mouth opening restrictions in patients with PTSD by means of splint therapy (st) show long-term therapeutic effects in the case of functional disorders. MATERIAL AND METHODS: In 31 of 36 inpatients (soldiers, average age 37.1 ± 7.3 years, 26.7 ± 2.1 teeth) with confirmed posttraumatic stress disorder, chronic pain intensity > 6 (visual analogue scale 0 to 10), the mouth opening was determined, and the functional status (RDC-TMD) was recorded. All participants received a splint that was worn at night. A control of the therapeutic effect of the splint occurred after 6 weeks, 3, 6, and 12 months. RESULTS: The mouth opening initially had an average of 30.9 ± 6.5 mm (median 31 mm). The pain intensity (PI) was reported to be on average VAS 8.3 ± 0.9, the chronic degree of pain according to von Korff was 3.9 ± 03. Six weeks after the st (n = 31), the average mouth opening was 49.5 ± 6.3 mm (median 51.5). PI was given as VAS 2.3 ± 1.1 on average. After 3, 6, and 12 months, 24, 15, and 14 subjects could be interviewed regarding PI. Based on the last examination date of all subjects, the average PI was given as 1.1 ± 0.9 (median 1). CONCLUSION: The presented data show that the therapeutic short-term results achieved by means of a splint remain valid on the long term despite continued PTSD. CLINICAL RELEVANCE: The presented study shows that patients will benefit in the long term from a splint and remain symptom-free, even if this mental illness persists.

Obesity-induced changes in lipid mediators persist after weight loss.

BACKGROUND: Obesity induces significant changes in lipid mediators, however, the extent to which these changes persist after weight loss has not been investigated. SUBJECTS/METHODS: We fed C57BL6 mice a high-fat diet to generate obesity and then switched the diet to a lower-fat diet to induce weight loss. We performed a comprehensive metabolic profiling of lipid mediators including oxylipins, endocannabinoids, sphingosines and ceramides in key metabolic tissues (including adipose, liver, muscle and hypothalamus) and plasma. RESULTS: We found that changes induced by obesity were largely reversible in most metabolic tissues but the adipose tissue retained a persistent obese metabolic signature. Prostaglandin signaling was perturbed in the obese state and lasting increases in PGD2, and downstream metabolites 15-deoxy PGJ2 and delta-12-PGJ2 were observed after weight loss. Furthermore expression of the enzyme responsible for PGD2 synthesis (hematopoietic prostaglandin D synthase, HPGDS) was increased in obese adipose tissues and remained high after weight loss. We found that inhibition of HPGDS over the course of 5 days resulted in decreased food intake in mice. Increased HPGDS expression was also observed in human adipose tissues obtained from obese compared with lean individuals. We then measured circulating levels of PGD2 in obese patients before and after weight loss and found that while elevated relative to lean subjects, levels of this metabolite did not decrease after significant weight loss. CONCLUSIONS: These results suggest that lasting changes in lipid mediators induced by obesity, still present after weight loss, may play a role in the biological drive to regain weight.

Effects of combined helium pre/post-conditioning on the brain and heart in a rat resuscitation model.

BACKGROUND: The noble gas helium induces cardio- and neuroprotection by pre- and post-conditioning. We investigated the effects of helium pre- and post-conditioning on the brain and heart in a rat resuscitation model. METHODS: After approval by the Animal Care Committee, 96 Wistar rats underwent cardiac arrest for 6 min induced by ventricular fibrillation. Animals received 70% helium and 30% oxygen for 5 min before cardiac arrest and for 30 min after restoration of spontaneous circulation (ROSC). Control animals received 70% nitrogen and 30% oxygen. Hearts and brains were excised after 2, 4 h or 7 days. Neurological degeneration was evaluated using TUNEL and Nissl staining in the hippocampal CA-1 sector. Cognitive function after 7 days was detected with the tape removal test. Molecular targets were measured by infrared western blot. Data are shown as median [Interquartile range]. RESULTS: Helium treatment resulted in significantly less apoptosis (TUNEL positive cells/100 pixel 73.5 [60.3-78.6] vs.78.2 [70.4-92.9] P = 0.023). Changes in Caveolin-3 expression in the membrane fraction and Hexokinase-II in the mitochondrial fraction were observed in the heart. Caveolin-1 expression of treated animals significantly differed from control animals in the membrane fraction of the heart and brain after ROSC. CONCLUSION: Treatment with helium reduced apoptosis in our resuscitation model. Differential expression levels of Caveolin-1, Caveolin-3 and Hexokinase II in the heart were found after helium pre- and post-conditioning. No beneficial effects were seen on neurofunctional outcome.

Life-history strategies of the rock hind grouper Epinephelus adscensionis at Ascension Island.

Epinephelus adscensionis sampled from Ascension Island, South Atlantic Ocean, exhibits distinct life-history traits, including larger maximum size and size at sexual maturity than previous studies have demonstrated for this species in other locations. Otolith analysis yielded a maximum estimated age of 25 years, with calculated von Bertalanffy growth parameters of: L∞ = 55·14, K = 0·19, t0 = -0·88. Monthly gonad staging and analysis of gonad-somatic index (IG ) provide evidence for spawning from July to November with an IG peak in August (austral winter), during which time somatic growth is also suppressed. Observed patterns of sexual development were supportive of protogyny, although further work is needed to confirm this. Mean size at sexual maturity for females was 28·9 cm total length (LT ; 95% C.I. 27·1-30·7 cm) and no females were found >12 years and 48·0 cm LT , whereas all confirmed males sampled were mature, >35·1 cm LT with an age range from 3 to 18 years. The modelled size at which 50% of individuals were male was 41·8 cm (95% C.I. 40·4-43·2 cm). As far as is known, this study represents the first comprehensive investigation into the growth and reproduction of E. adscensionis at its type locality of Ascension Island and suggests that the population may be affected less by fisheries than elsewhere in its range. Nevertheless, improved regulation of the recreational fishery and sustained monitoring of abundance, length frequencies and life-history parameters are needed to inform long-term management measures, which could include the creation of marine reserves, size or temporal catch limits and stricter export controls.

Management of systemic AL amyloidosis: recommendations of the Myeloma Foundation of Australia Medical and Scientific Advisory Group.

Systemic AL amyloidosis is a plasma cell dyscrasia with a characteristic clinical phenotype caused by multi-organ deposition of an amyloidogenic monoclonal protein. This condition poses a unique management challenge due to the complexity of the clinical presentation and the narrow therapeutic window of available therapies. Improved appreciation of the need for risk stratification, standardised use of sensitive laboratory testing for monitoring disease response, vigilant supportive care and the availability of newer agents with more favourable toxicity profiles have contributed to the improvement in treatment-related mortality and overall survival seen over the past decade. Nonetheless, with respect to the optimal management approach, there is a paucity of high-level clinical evidence due to the rarity of the disease, and enrollment in clinical trials is still the preferred approach where available. This review will summarise the Clinical Practice Guidelines on the Management of Systemic Light Chain (AL) Amyloidosis recently prepared by the Medical Scientific Advisory Group of the Myeloma Foundation of Australia. It is hoped that these guidelines will assist clinicians in better understanding and optimising the management of this difficult disease.

Selection of hospital antimicrobial prescribing quality indicators: a consensus among German antibiotic stewardship (ABS) networkers.

PURPOSE: Simple, valid, and evidence-based indicators to measure the quality of antimicrobial prescribing in acute-care hospitals are urgently needed and increasingly requested by policymakers. The aim of this study was to develop new consensus quality indicators (QIs) for hospital antibiotic stewardship (ABS) and infection management which will be further evaluated for internal quality management and external quality assessment in Germany. METHODS: Based on an extensive literature review, the Austrian-German hospital ABS Guideline Committee and selected members of the German ABS Expert Network discussed and drafted a list of 99 potential indicators for hospitals that reflect structural prerequisites for ABS (35 items), ABS core activities (18 items), additional ABS measures (5 items), and process of care indicators (both generic and disease-specific-12 and 29 items, respectively). Questionnaires were mailed to German ABS experts and healthcare professionals with further education in ABS. Participants scored (on a nine-point Likert scale) relevance (clinical, ecological/resistance, economical/expenses) and presumed practicability (six categories: clarity of definition, effort to collect data, barrier to implementation, verifiability, suitability for external quality assessment, quality gap), taking into account their local work environment. The scores were processed according to the RAND/UCLA appropriateness method, and QIs were judged relevant if the median (clinical + ecological and/or economical) scores were >6. The indicators thus assessed to be potentially relevant were then filtered according to their practicability. Highly relevant QIs with borderline practicability scores and items with disagreements and overlapping areas were re-discussed in a final multidisciplinary panel consensus workshop convened in November 2012. RESULTS: Of the 340 questionnaires that were mailed, 75 questionnaires were completed and returned. Of 99 initially proposed items, 32 were excluded due to insufficient scores. Of the remaining 67 items, 21 structural and 21 process of care QIs were finally selected, including four QIs with high clinical and ecological but limited economical relevance, and three QIs with high clinical and economical but limited ecological relevance. Among the selected QIs, efforts to collect data and implementation barriers were scored as suboptimal in many cases. CONCLUSIONS: A catalog of consensus structural and process of care ABS-QIs was established. These should undergo further pilot and feasibility studies in the German hospital healthcare sector. The panelists were most critical regarding resource use/complexity issues and presumed implementation barriers. How this may limit applicability of QIs remains to be determined.

Lidocaine and ropivacaine, but not bupivacaine, demethylate deoxyribonucleic acid in breast cancer cells in vitro.

BACKGROUND: Lidocaine demethylates deoxyribonucleic acid (DNA) in breast cancer cells. This modification of epigenetic information may be of therapeutic relevance in the perioperative period, because a decrease in methylation can reactivate tumour suppressor genes and inhibit tumour growth. The objectives of this study were to determine the effect of two amide local anaesthetics, ropivacaine and bupivacaine, on methylation in two breast cancer cell lines and to detect whether the combination of lidocaine with the chemotherapy agent 5-aza-2'-deoxycytidine (DAC) would result in additive demethylating effects. METHODS: Breast cancer cell lines BT-20 [oestrogen receptor (ER)-negative] and MCF-7 (ER-positive) were incubated with lidocaine, bupivacaine, and ropivacaine to assess demethylating properties. Then, we tested varying concentrations of lidocaine and DAC to assess whether their demethylating effects were additive. Cell numbers and global methylation status were analysed. RESULTS: Lidocaine decreased methylation in BT-20 and MCF-7 cells, ropivacaine decreased methylation in BT-20 cells, and bupivacaine had no demethylating effect. When combined, lidocaine and DAC had additive demethylating effects. CONCLUSIONS: At clinically relevant doses, lidocaine and ropivacaine exert demethylating effects on specific breast cancer cell lines, but bupivacaine does not. The demethylating effects of lidocaine and DAC are indeed additive.

Patterns of direct and indirect contact between cattle and badgers naturally infected with tuberculosis.

Tuberculosis (TB) due to infection with Mycobacterium bovis is transmitted between cattle and badgers (Meles meles) in the UK and Ireland but it is unclear where or when transmission occurs. We investigated direct and indirect interactions between badgers and cattle using automated proximity loggers on animals and at badger latrines located on pasture, in an area of south-west England with a high-density badger population. Direct contacts (interactions within 1.4 m) between badgers and cattle at pasture were very rare (four out of >500000 recorded animal-to-animal contacts) despite ample opportunity for interactions to occur. Indirect interactions (visits to badger latrines by badgers and cattle) were two orders of magnitude more frequent than direct contacts: 400 visits by badgers and 1700 visits by cattle were recorded. This suggests that indirect contacts might be more important than direct contacts in terms of disease transmission at pasture. The TB infection status of individual badgers (ascribed with 93% accuracy using three diagnostic tests) did not affect the frequency or duration of their visits to latrines located on pasture grazed by cattle. Nevertheless, there was wide variation in contact behaviour between individuals, which highlights the importance of understanding heterogeneity in contact patterns when developing strategies to control disease spread in wildlife and livestock.

2D imaging spin-filter for NanoESCA based on Au/Ir(001) or Fe(001)-p(1×1)O.

A two-dimensional imaging spin-filter for photo-emission electron microscopy is described. The spin-filter is capable of imaging the electron spin polarization of either real space or momentum space electron distributions. As a scattering target either Au/Ir(001) comes into use, where spin sensitivity results from using spin-orbit scattering or Fe(001)-p(1×1)O that exploits exchange interaction. Both scattering targets were characterized with respect to their working points and Sherman function in a separate setup. Spin-polarization images of secondary electrons from the magnetic domains of a poly-crystalline iron sample are shown using both scattering targets. Images with a spin-filter using Au/Ir(001) show more than 104 discrete detection channels which increases the effective two-dimensional figure-of-merit (FoM) of this spin-filter by four orders of magnitude compared to single-channel spin detectors. Using the exchange scattering target two spin-components have been imaged for the first time. A method to detect all three spin-components is also outlined.

[Giant hamartoma of the lung]. / Riesiges Hamartom der Lunge.

Hamartomas are the most common benign tumors of the lungs and usually present as solitary intraparenchymal lesions a few centimeters in diameter which are detected radiologically as an incidental finding. This article reports on a case of a giant pulmonary hamartochondroma 29 cm in diameter, which impressed preoperatively and intraoperatively as an extrapulmonary neoplasm. The detection of smaller mesenchymal neoplastic lesions in the lungs and pleura is a further special feature of this case.

A New Frontier in Temporal Bone Imaging: Photon-Counting Detector CT Demonstrates Superior Visualization of Critical Anatomic Structures at Reduced Radiation Dose.

BACKGROUND AND PURPOSE: Photon-counting detector CT is a new technology with a limiting spatial resolution of ≤150 µm. In vivo comparisons between photon-counting detector CT and conventional energy-integrating detector CT are needed to determine the clinical impact of photon counting-detector CT in temporal bone imaging. MATERIALS AND METHODS: Prospectively recruited patients underwent temporal bone CT examinations on an investigational photon-counting detector CT system after clinically indicated temporal bone energy-integrating detector CT. Photon-counting detector CT images were obtained at an average 31% lower dose compared with those obtained on the energy-integrating detector CT scanner. Reconstructed images were evaluated in axial, coronal, and Pöschl planes using the smallest available section thickness on each system (0.4 mm on energy-integrating detector CT; 0.2 mm on photon-counting detector CT). Two blinded neuroradiologists compared images side-by-side and scored them using a 5-point Likert scale. A post hoc reassignment of readers' scores was performed so that the scores reflected photon-counting detector CT performance relative to energy-integrating detector CT. RESULTS: Thirteen patients were enrolled, resulting in 26 image sets (left and right sides). The average patient age was 63.6 [SD, 13.4] years; 7 were women. Images from the photon-counting detector CT scanner were significantly preferred by the readers in all reconstructed planes (P < .001). Photon-counting detector CT was rated superior for the evaluation of all individual anatomic structures, with the oval window (4.79) and incudostapedial joint (4.75) receiving the highest scores on a Likert scale of 1-5. CONCLUSIONS: Temporal bone CT images obtained on a photon-counting detector CT scanner were rated as having superior spatial resolution and better critical structure visualization than those obtained on a conventional energy-integrating detector scanner, even with a substantial dose reduction.

[Biopsy diagnosis of papillary adenoma of the lung by endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA)]. / Bioptische Diagnose eines papillären Adenoms der Lunge mittels Ultraschall-gesteuerter transbronchialer Nadelaspiration (EBUS-TBNA).

The papillary adenoma of the lung is a rare benign neoplasia with intraparenchymal localisation and is thus, in general, difficult to access for biopsy diagnosis. Due to the rarity of this lesion, the intra-operative histological diagnosis by frozen section may be problematic. We report on a 75-year-old male patient with a lung nodule, biopsied by means of bronchoscopy with TBNA. The tissue obtained was processed histologically as a cell block and examined microscopically. The diagnosis of a papillary adenoma was made. Our case is the first report on the diagnosis of a papillary adenoma of the lung by TBNA biopsy. For our patient with impaired lung function the diagnostic procedure described here represented a less invasive alternative to the diagnostic evaluation by surgery with intraoperative frozen section examination. This case shows that the TBNA biopsy combined with processing of the tissue as a cell block may represent a contribution to the appropriate management of the patients and serve as an aid for therapy planning.

Low protein diets without medicinal zinc oxide for weaned pigs reduced diarrhea treatments and average daily gain.

The use of medicinal zinc oxide (ZnO) in post-weaning diets must be phased out in the European Union by 2022, resulting in urgent needs for alternative strategies to prevent diarrhea in pigs. The objective of this study was to test the effect of four different dietary protein strategies with different amino acid profiles on diarrhea frequency and pig performance as alternative diet plans when medicinal ZnO was will be excluded from the diet. A total of 6 800 Duroc x (Danish Landrace x Yorkshire) pigs, weaned around 28 days of age, were randomly assigned by sex and size to six dietary treatments. The treatments were; standard CP levels (191, 184, 184 g/kg CP) and allocated 2 500 ppm ZnO in phase 1 (PC = positive control), standard CP levels (191, 184, 184 g/kg CP) and no added ZnO in phase 1 (NC = negative control), CP levels of 166, 184, 184 g/kg (LSS = low-standard-standard), CP levels of 166, 162, 192 g/kg (LLH = low-low-high), CP levels of 140, 193, 192 g/kg (VHH = very low-high-high), and lastly 140, 174, 192 g/kg CP levels (VMH = very low-medium-high). The pigs entered the trial at ~7 kg BW and exited at ~30 kg BW. As expected, the PC treatment resulted in 42% fewer diarrhea pen treatments in the total trial period compared to the NC group (P < 0.05), whereas both PC and LLH had fewer diarrhea treatment days per pig compared to NC pigs (P < 0.05). Additionally, at the point of treatment, the NC pens had 33% fecal floor samples positive for pathogens, compared to 80% samples positive for pathogens in the PC pens. This suggests that ZnO has a particular positive effect on non-infectious diarrhea without bacterial involvement. A reduction in dietary CP levels in phase 1 led to a reduced average daily gain (ADG) in LLH and VMH pigs and a poorer feed conversion ratio (FCR) in VHH pigs during the overall study period compared to the NC pigs (P < 0.05). Conclusively, a diet with low CP levels from weaning to about 15 kg BW had a reducing effect on diarrhea, but decreased ADG without affecting the FCR.