Search Results for: Probiotici: in vivo vs in vitro

Objectives
Intensive care unit (ICU) clinicians stop antibiotics more often, with a negative infection: point-of-care test (PCR-POCT). Simulated cases of diagnostic uncertainty regarding infection resolution led clinicians to choose options such as procalcitonin (PCT) and/or PCR-POCTs +/– de-escalation to aid stop decisions. We hypothesised that a direct infection indicator, PCR-POCT, would influence stop judgements more than indirect PCT. Accordingly, we tested antibiotic-stop decisions when presented with a negative PCR-POCT despite borderline-positive PCT.

Designs
Observational prospective study.

Setting
ICU.

Participants
66 ICU clinicians from University hospitals.

Methods
Clinicians saw four scenarios of different clinico-biological trajectories: (1) clear improvement, (2) clear worsening, (3) discordant—clinically better/biologically worse and (4) discordant—clinically worse/biologically better. Participants gave an initial decision (stop/continue/continue–escalate/continue–de-escalate). Then PCR-POCT and/or PCT was offered (accept/decline). After a negative PCR-POCT and borderline-positive PCT result, a final antibiotic decision was taken.

Measures
Proportion of stop decisions before versus after test results per scenario. The association of the final decision with the clinician’s change in confidence, willingness to request the biomarker(s) and the case trajectory was determined.

Results
Fewer clinicians than expected stopped antibiotics versus baseline (36%, 94/264 vs 42%, 110/264, p=0.045). This was so in three of four scenarios, significantly less in the improvement (p

Objective
To assess the effectiveness of a potent topical corticosteroid (TCS) as an initial treatment in primary care for children with moderate flare-ups of atopic dermatitis (AD), compared to starting on a mild TCS.

Design
An observational prospective cohort study with an embedded pragmatic multicentre open-label randomised controlled trial.

Setting
A total of 53 general practices in the southwest of the Netherlands took part in the study.

Participants
209 children aged 3 months to 17 years diagnosed with AD (International Classification of Primary Care codes S87 or S88) who visited their general practitioner (GP) for AD or received repeat prescriptions for AD in the previous 12 months were included in the cohort study through the general practices. Finally, 32 patients (15%) were randomised and assigned to the trial (13 girls; 19 boys; median age 4.0 years).

Interventions
If cohort participants experienced a moderate flare-up (ie, need to intensify topical treatment from the child’s and/or parents’ point of view of AD and a three-item severity score from three to

Circulation, Ahead of Print. BACKGROUND:Cardiac fibrosis, characterized by excessive extracellular matrix (ECM) deposition in the myocardium, is an important target for heart disease treatments.Pw1(paternally expressed gene 3) is an imprinted gene expressed from the paternal allele, and de novo purine biosynthesis (DNPB) is a crucial pathway for nucleotide synthesis. However, the roles of PW1 and DNPB in ECM production by cardiac fibroblasts during myocardial ischemia are not yet understood.METHODS:To induce myocardial damage, we performed left anterior descending coronary artery ligation. We generatedPw1CreER-2A-eGFPandPw12A-CreERknock-in mouse lines to evaluate the expression of the 2Pw1alleles in normal and injured hearts. Bisulfite sequencing was used to analyze the DNA methylation of thePw1imprinting control region. We identified the phosphoribosylformylglycinamidine synthase (Pfas) gene, encoding the DNPB enzyme PFAS, as a direct target of PW1 using chromatin immunoprecipitation sequencing and real-time quantitative polymerase chain reaction. The role of DNPB in ECM production and cardiac fibrosis after injury was examined in vitro using cultured cardiac fibroblasts and in vivo withPfas-deficient mice.RESULTS:Our study demonstrates that myocardial infarction reduces DNA methylation at the imprinting control region of the maternally imprinted genePw1, triggering a switch from monoallelic imprinting to biallelic expression ofPw1in cardiac fibroblasts. In activated cardiac fibroblasts, increasedPw1expression promotes purine biosynthesis and induces ECM production by transcriptionally activating the DNPB factorPfas. We identified that DNPB is essential for ECM production in activated fibroblasts and that loss ofPfasin fibroblasts limits cardiac fibrosis and improves heart function after injury.CONCLUSIONS:This study demonstrates thatPw1imprinting is disrupted after injury and reveals a novel role for the downstream target PFAS in ECM production and cardiac fibrogenesis. Targeting the PW1/PFAS signaling pathway presents a promising therapeutic strategy for improving cardiac repair after injury.

Stroke, Volume 56, Issue 1, Page 170-182, January 1, 2025. BACKGROUND:Ischemic stroke is a common cause of death worldwide and a main cause of morbidity. Presently, laser speckle contrast imaging, x-ray computed tomography, and magnetic resonance imaging are the mainstay for stroke diagnosis and therapeutic monitoring in preclinical studies. These modalities are often limited in terms of their ability to map brain perfusion with sufficient spatial and temporal resolution, thus calling for development of new brain perfusion techniques featuring rapid imaging speed, cost-effectiveness, and ease of use.METHODS:We report on a new preclinical high-resolution angiography technique for murine ischemic stroke imaging based on large-field high-speed multifocal illumination fluorescence microscopy. We subsequently showcase the proposed method by monitoring therapeutic effects of thrombolysis in stroke (n=6), further performing cross-strain comparison of perfusion dynamics (n=6) and monitoring the therapeutic effects of sensory stimulation–based treatment (n=11).RESULTS:Quantitative readings of hemodynamic and structural changes in cerebral vascular network and pial vessels were attained with 14.4-µm spatial resolution at 80-Hz frame rate fully transcranially. The in vivo perfusion maps accurately delineated the ischemic core and penumbra, further exhibiting a strong correlation (86.1±4.5%) with ex vivo triphenyl tetrazolium chloride staining, significantly higher than for the conventional laser speckle contrast imaging method. Monitoring of therapeutic effects of thrombolysis confirmed that early recanalization could effectively save the penumbra while reducing the infarct area. Cross-strain comparison of perfusion dynamics affirmed that C57BL/6 mice feature a larger penumbra and smaller infarct core as compared with BALB/c mice, which have few or no collaterals. Sensory stimulation–based treatment could effectively enhance blood flow and abolish perfusion deficits in the ischemic core and penumbra regions.CONCLUSIONS:A high-speed fluorescence-based angiography method for transcranial stroke imaging in mice is introduced, which is capable of localizing brain perfusion changes and accurately assessing the ischemic penumbra. Compared with the whole-brain x-ray computed tomography and magnetic resonance imaging methods, which are conventionally used for stroke diagnosis and therapeutic monitoring, the new approach is simple and cost-effective, further offering high resolution and speed for in vivo studies. It thus opens new venues for brain perfusion research under various disease conditions such as stroke, neurodegeneration, or epileptic seizures.

Portal vein thromboses (PVTs) are common in patients with cirrhosis and are associated with advanced portal hypertension and mortality. The treatment of PVTs remains a clinical challenge due to limited evidence and competing risks of PVT-associated complications vs bleeding risk of anticoagulation. Significant heterogeneity in PVT phenotype based on anatomic, host, and disease characteristics, and an emerging spectrum of therapeutic options further complicate PVT management. This Clinical Practice Update (CPU) aims to provide best practice advice for the evaluation and management of PVT in cirrhosis, including the role of direct oral anticoagulants and endovascular interventions.

This narrative review explores treatment with surgical aortic valve replacement vs transcatheter aortic valve implantation in older adults with severe symptomatic calcific aortic stenosis, including the type and timing of valve replacement.

Objective
We compared processes of antepartum, intrapartum and postpartum care and obstetrical outcomes between physicians and non-physicians.

Design
This is a population-based retrospective matched cohort study.

Setting
The study was conducted in Ontario, Canada.

Participants
Physicians and non-physicians residing in high-income urban areas from 1 April 2009 to 26 November 2018 were included. Physicians were matched to non-physicians on maternal age, calendar year, parity, conception by assisted reproductive technology and singleton versus multifetal gestation. We compared processes of antepartum, intrapartum and postpartum care between physicians and non-physicians.

Outcome measures
The primary outcome was mode of delivery (caesarean section, C-section vs vaginal delivery). Secondary outcomes included obstetrical anal sphincter injury among those experiencing vaginal birth and differences in urgent healthcare contacts (maternal and neonatal) during the postpartum period.

Results
7327 physicians were matched 1:5 to 36 185 non-physicians and were well balanced except for comorbidities (physicians had fewer comorbidities). Physicians had more antenatal ultrasounds and invasive prenatal testing, received labour anaesthesia more often and were more often delivered by their own care provider. In adjusted analyses, physicians and non-physicians had a similar risk of C-section (aRR 0.97, 95% CI 0.93 to 1.00, p=0.07). There was no difference in neonatal urgent care contacts; non-physicians had a higher risk of maternal urgent postpartum care (adjusted relative risk [aRR] 1.22, 95% CI 1.08 to 1.37, p

Objectives
Recent studies have suggested a potential link between opium consumption and microvascular dysfunction in coronary arteries, which may contribute to the development of coronary slow-flow syndrome. This study aims to investigate the relationship between opium use and coronary slow-flow syndrome.

Design and setting
This retrospective study analysed medical records of patients who underwent coronary angiography at the Tehran Heart Center from 2006 to 2020. It focused on those with coronary slow flow phenomenon (CSFP) or non-obstructive coronary artery disease, excluding patients with significant left ventricular dysfunction (left ventricular ejection fraction

Objective
To analyse the effects of tracheostomy timing on COVID-19 outcomes by comparing mortality rates at different time points (7, 10 and 14 days).

Design
Systematic review and meta-analysis.

Data sources
PubMed, Embase, Cochrane Library, Web of Science and Scopus were searched from 31 August 2023 to 6 September 2023.

Primary and secondary outcomes measures
The primary outcome was short-term mortality, defined as intensive care unit (ICU) mortality, hospital mortality and 28-day or 30-day mortality. The secondary outcomes included mechanical ventilation duration, ICU and hospital days.

Results
Among 3465 patients from 12 studies, the 10-day subgroup analysis revealed higher mortality for earlier tracheostomy than for later tracheostomy (49.7% vs 32.6%, OR 1.91, 95% CI 1.37–2.65). No significant differences were observed at 7- and 14-day marks. Earlier tracheostomy was associated with shorter mechanical ventilation (mean difference=–7.35 days, 95% CI –11.63 to –0.38) and ICU stays (mean difference=–11.24 days, 95% CI –18.50 to –3.97) compared with later tracheostomy. Regarding hospital stay, the later tracheostomy group exhibited a trend towards longer-term inpatients, with no significant difference.

Conclusions
No significant difference in short-term mortality was observed between patients undergoing tracheostomy at 7 and 14 days; however, at 10 days, later tracheostomy resulted in a lower mortality rate. Accordingly, subtle timing differences may impact short-term results in COVID-19 patients. Considering that the later tracheostomy group had longer mechanical ventilation and ICU stays, additional research is required to determine an optimal timing that reduces mortality cost-effectively.

Objective
Sphincter of Oddi disorders (SOD) are contentious conditions in patients whose abdominal pain, idiopathic acute pancreatitis (iAP) might arise from pressurisation at the sphincter of Oddi. The present study aimed to measure the benefit of sphincterotomy for suspected SOD.

Design
Prospective cohort conducted at 14 US centres with 12 months follow-up. Patients undergoing first-time endoscopic retrograde cholangiopancreatography (ERCP) with sphincterotomy for suspected SOD were eligible: pancreatobiliary-type pain with or without iAP. The primary outcome was defined as the composite of improvement by Patient Global Impression of Change (PGIC), no new or increased opioids and no repeat intervention. Missing data were addressed by hierarchal, multiple imputation scheme.

Results
Of 316 screened, 213 were enrolled with 190 (89.2%) of these having a dilated bile duct, abnormal labs, iAP or some combination. By imputation, an average of 122/213 (57.4% (95% CI 50.4% to 64.4%)) improved; response rate was similar for those with complete follow-up (99/161, 61.5% (54.0% to 69.0%)); of these, 118 (73.3%) improved by PGIC alone. Duct size, elevated labs and patient characteristics were not associated with response. AP occurred in 37/213 (17.4%) at a median of 6 months post ERCP and was more likely in those with a history of AP (30.9% vs 2.9%, p

Background
Colonic motility in constipation can be assessed non-invasively using MRI.

Objective
To compare MRI with high-resolution colonic manometry (HRCM) for predicting treatment response.

Design
Part 1: 44 healthy volunteers (HVs), 43 patients with irritable bowel syndrome with constipation (IBS-C) and 37 with functional constipation (FC) completed stool diaries and questionnaires and underwent oral macrogol (500–1000 mL) challenge. Whole gut transit time (WGTT), segmental colonic volumes (CV), MRI-derived Motility Index and chyme movement by ‘tagging’ were assessed using MRI and time to defecation after macrogol recorded. Left colonic HRCM was recorded before and after a 700 kcal meal. Patients then proceeded to Part 2: a randomised cross-over study of 10-days bisacodyl 10 mg daily versus hyoscine 20 mg three times per day, assessing daily pain and constipation.

Results
Part 1: Total CVs median (range) were significantly greater in IBS-C (776 (595–1033)) and FC (802 (633–951)) vs HV (645 (467–780)), p

Background
The potency of T cell-mediated responses is a determinant of immunotherapy effectiveness in treating malignancies; however, the clinical efficacy of T-cell therapies has been limited in hepatocellular carcinoma (HCC) owing to the extensive immunosuppressive microenvironment.

Objective
Here, we aimed to investigate the key genes contributing to immune escape in HCC and raise a new therapeutic strategy for remoulding the HCC microenvironment.

Design
The genome-wide in vivo clustered regularly interspaced short palindromic repeats (CRISPR) screen library was conducted to identify the key genes associated with immune tolerance. Single-cell RNA-seq (scRNA-seq), flow cytometry, HCC mouse models, chromatin immunoprecipitation and coimmunoprecipitation were used to explore the function and mechanism of adenylate cyclase 7 (ADCY7) in HCC immune surveillance.

Results
Here, a genome-wide in vivo CRISPR screen identified a novel immune modulator-ADCY7. The transmembrane protein ADCY7 undergoes subcellular translocation via caveolae-mediated endocytosis and then translocates to the nucleus with the help of leucine-rich repeat-containing protein 59 (LRRC59) and karyopherin subunit beta 1 (KPNB1). In the nucleus, it functions as a transcription cofactor of CCAAT/enhancer binding protein alpha (CEBPA) to induce CCL5 transcription, thereby increasing CD8+ T cell infiltration to restrain HCC progression. Furthermore, ADCY7 can be secreted as exosomes and enter neighbouring tumour cells to promote CCL5 induction. Exosomes with high ADCY7 levels promote intratumoural infiltration of CD8+ T cells and suppress HCC tumour growth.

Conclusion
We delineate the unconventional function and subcellular location of ADCY7, highlighting its pivotal role in T cell-mediated immunity in HCC and its potential as a promising treatment target.

Background and aims
The efficacy of colorectal endoscopic mucosal resection (EMR) is limited by recurrence and the necessity for conservative surveillance. Margin thermal ablation (MTA) after EMR has reduced the incidence of recurrence at the first surveillance colonoscopy at 6 months (SC1). Whether this effect is durable to second surveillance colonoscopy (SC2) is unknown. We evaluated long-term surveillance outcomes in a cohort of LNPCPs that have undergone MTA.

Methods
LNPCPs undergoing EMR and MTA from four academic endoscopy centres were prospectively recruited. EMR scars were evaluated at SC1 and in the absence of recurrence, SC2 colonoscopy was conducted in a further 12 months. A historical control arm was generated from LNPCPs that underwent EMR without MTA. The primary outcome was recurrence at SC2 in all LNPCPs with a recurrence-free scar at SC1.

Results
1152 LNPCPs underwent EMR with complete MTA over 90 months until October 2022. 854 LNPCPs underwent SC1 with 29/854 (3.4%) LNPCPs demonstrating recurrence. 472 LNPCPs free of recurrence at SC1 underwent SC2. 260 LNPCPs with complete SC2 follow-up formed the control arm from January 2012 to May 2016. Recurrence at SC2 was significantly less in the MTA arm versus controls (1/472 (0.2%) vs 9/260 (3.5%); p

Objective
Fat mass and obesity-associated protein (FTO), an eraser of N 6-methyadenosine (m6A), plays oncogenic roles in various cancers. However, its role in hepatocellular carcinoma (HCC) is unclear. Furthermore, small extracellular vesicles (sEVs, or exosomes) are critical mediators of tumourigenesis and metastasis, but the relationship between FTO-mediated m6A modification and sEVs in HCC is unknown.

Design
The functions and mechanisms of FTO and glycoprotein non-metastatic melanoma protein B (GPNMB) in HCC progression were investigated in vitro and in vivo. Neutralising antibody of syndecan-4 (SDC4) was used to assess the significance of sEV-GPNMB. FTO inhibitor CS2 was used to examine the effects on anti-PD-1 and sorafenib treatment.

Results
FTO expression was upregulated in patient HCC tumours. Functionally, FTO promoted HCC cell proliferation, migration and invasion in vitro, and tumour growth and metastasis in vivo. FTO knockdown enhanced the activation and recruitment of tumour-infiltrating CD8+ T cells. Furthermore, we identified GPNMB to be a downstream target of FTO, which reduced the m6A abundance of GPNMB, hence, stabilising it from degradation by YTH N 6-methyladenosine RNA binding protein F2. Of note, GPNMB was packaged into sEVs derived from HCC cells and bound to the surface receptor SDC4 of CD8+ T cells, resulting in the inhibition of CD8+ T cell activation. A potential FTO inhibitor, CS2, suppresses the oncogenic functions of HCC cells and enhances the sensitivity of anti-PD-1 and sorafenib treatment.

Conclusion
Targeting the FTO/m6A/GPNMB axis could significantly suppress tumour growth and metastasis, and enhance immune activation, highlighting the potential of targeting FTO signalling with effective inhibitors for HCC therapy.

Circulation, Ahead of Print. BACKGROUND:Ca2+mishandling in cardiac Purkinje cells is a well-known cause of cardiac arrhythmias. The Purkinje cell resident inositol 1,4,5-trisphosphate receptor 1 (ITPR1) is believed to play an important role in Ca2+handling, and ITPR1 gain-of-function (GOF) has been implicated in cardiac arrhythmias. However, nearly all known disease-associated ITPR1 variants are loss-of-function and are primarily linked to neurological disorders. Whether ITPR1 GOF has pathological consequences, such as cardiac arrhythmias, is unclear. This study aimed to identify human ITPR1 GOF variants and determine the impact of ITPR1 GOF on Ca2+handling and arrhythmia susceptibility.METHODS:There are a large number of rare ITPR1 missense variants reported in open data repositories. Based on their locations in the ITPR1 channel structure, we selected and characterized 33 human ITPR1 missense variants from open databases and identified 21 human ITPR1 GOF variants. We generated a mouse model carrying a human ITPR1 GOF variant, ITPR1-W1457G (W1447G in mice).RESULTS:We showed that the ITPR1-W1447G±and recently reported ITPR1-D2594K±GOF mutant mice were susceptible to stress-induced ventricular arrhythmias. Confocal Ca2+and voltage imaging in situ in heart slices and Ca2+imaging and patch-clamp recordings of isolated Purkinje cells showed that ITPR1-W1447G±and ITPR1-D2594K±variants increased the occurrence of stress-induced spontaneous Ca2+release, delayed afterdepolarization, and triggered activity in Purkinje cells. To assess the potential role of ITPR1 variants in arrhythmia susceptibility in humans, we looked up a gene-based association study in the UK Biobank data set and identified 7 rare ITPR1 missense variants showing potential association with cardiac arrhythmias. Remarkably, in vitro functional characterization revealed that all these 7 ITPR1 variants resulted in GOF.CONCLUSIONS:Our studies in mice and humans reveal that enhanced function of ITPR1, a well-known movement disorder gene, increases the risk for cardiac arrhythmias.

This Viewpoint discusses enrollment in Medicare Advantage vs traditional Medicare among older adults and common reasons for plan disenrollment, including the lack of in-network physicians and hospitals.