Intelligent Liver Function Testing: Working Smarter to Improve Patient Outcomes in Liver Disease.

Chronic liver disease (CLD) is a significant health problem affecting millions of people worldwide. In Scotland, CLD is a major cause of premature mortality. Liver function tests (LFTs) are a panel of frequently requested blood tests which may indicate liver disease. However, LFTs commonly contain at least one abnormal result, and abnormalities are rarely investigated to the extent recommended by national guidelines. The intelligent Liver Function Testing (iLFT) pathway is a novel, automated system designed to improve early diagnosis of liver disease. Initial abnormal LFT results trigger a cascade of reflexive testing to help identify the cause of any liver dysfunction. Algorithms combine these results with demographic and clinical data (such as patient age, body mass index, and alcohol intake) and fibrosis estimates to produce an electronic diagnosis and management plan. The pilot trial demonstrated that iLFT increased diagnosis of liver disease whilst remaining cost-effective. As such, iLFT has been fully operational across our region (NHS Tayside, Scotland) since August 2018. In the first year, iLFT generated over 2000 diagnoses from 1824 patient samples with an abnormality in the initial LFTs. The majority of these patients could be safely managed in primary care. iLFT allows maximal value to be obtained from liver blood tests across biochemistry, virology, immunology, and hematology with only minor changes to working practices. 'Intelligent', algorithm-led testing pathways break down the barrier between clinical and laboratory medicine and offer solutions to many of the challenges experienced in modern healthcare systems.

Intelligent liver function testing (iLFT): A trial of automated diagnosis and staging of liver disease in primary care.

BACKGROUND & AIMS: Liver function tests (LFTs) are frequently requested blood tests which may indicate liver disease. LFTs are commonly abnormal, the causes of which can be complex and are frequently under investigated. This can lead to missed opportunities to diagnose and treat liver disease at an early stage. We developed an automated investigation algorithm, intelligent liver function testing (iLFT), with the aim of increasing the early diagnosis of liver disease in a cost-effective manner. METHODS: We developed an automated system that further investigated abnormal LFTs on initial testing samples to generate a probable diagnosis and management plan. We integrated this automated investigation algorithm into the laboratory management system, based on minimal diagnostic criteria, liver fibrosis estimation, and reflex testing for causes of liver disease. This algorithm then generated a diagnosis and/or management plan. A stepped-wedged trial design was utilised to compare LFT outcomes in general practices in the 6 months before and after introduction of the iLFT system. Diagnostic outcomes were collated and compared. RESULTS: Of eligible patients with abnormal LFTs, 490 were recruited to the control group and 64 were recruited to the intervention group. The primary diagnostic outcome was based on the general practitioner diagnosis, which agreed with the iLFT diagnosis in 67% of cases. In the iLFT group, the diagnosis of liver disease was increased by 43%. Additionally, there were significant increases in the rates of GP visits after diagnosis and the number of referrals to secondary care in the iLFT group. iLFT was cost-effective with a low initial incremental cost-effectiveness ratio of £284 per correct diagnosis, and a saving to the NHS of £3,216 per patient lifetime. CONCLUSIONS: iLFT increases liver disease diagnoses, improves quality of care, and is highly cost-effective. This can be achieved with minor changes to working practices and exploitation of functionality existing within modern laboratory diagnostics systems. LAY SUMMARY: There is a growing epidemic of advanced liver disease, this could be offset by early detection and management. Checking liver blood tests (LFTs) should be an opportunity to diagnose liver problems, but abnormal results are often incompletely investigated. In this study we were able to substantially increase the diagnostic yield of the abnormal LFTs using the automated intelligent LFT system. With the addition of referral recommendations and management plans, this strategy provides optimum investigation and management of LFTs and is cost saving to the NHS.

Correction to: Proteomic identification and characterization of hepatic glyoxalase 1 dysregulation in non-alcoholic fatty liver disease.

[This corrects the article DOI: 10.1186/s12953-018-0131-y.].

Proteomic identification and characterization of hepatic glyoxalase 1 dysregulation in non-alcoholic fatty liver disease.

Background: Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease worldwide. However, its molecular pathogenesis is incompletely characterized and clinical biomarkers remain scarce. The aims of these experiments were to identify and characterize liver protein alterations in an animal model of early, diet-related, liver injury and to assess novel candidate biomarkers in NAFLD patients. Methods: Liver membrane and cytosolic protein fractions from high fat fed apolipoprotein E knockout (ApoE-/-) animals were analyzed by quantitative proteomics, utilizing isobaric tags for relative and absolute quantitation (iTRAQ) combined with nano-liquid chromatography and tandem mass spectrometry (nLC-MS/MS). Differential protein expression was confirmed independently by immunoblotting and immunohistochemistry in both murine tissue and biopsies from paediatric NAFLD patients. Candidate biomarkers were analyzed by enzyme-linked immunosorbent assay in serum from adult NAFLD patients. Results: Through proteomic profiling, we identified decreased expression of hepatic glyoxalase 1 (GLO1) in a murine model. GLO1 protein expression was also found altered in tissue biopsies from paediatric NAFLD patients. In vitro experiments demonstrated that, in response to lipid loading in hepatocytes, GLO1 is first hyperacetylated then ubiquitinated and degraded, leading to an increase in reactive methylglyoxal. In a cohort of 59 biopsy-confirmed adult NAFLD patients, increased serum levels of the primary methylglyoxal-derived advanced glycation endproduct, hydroimidazolone (MG-H1) were significantly correlated with body mass index (r = 0.520, p < 0.0001). Conclusion: Collectively these results demonstrate the dysregulation of GLO1 in NAFLD and implicate the acetylation-ubquitination degradation pathway as the functional mechanism. Further investigation of the role of GLO1 in the molecular pathogenesis of NAFLD is warranted.

Genetic Factors Influencing Drug-Induced Liver Injury: Do They Have a Role in Prevention and Diagnosis?

PURPOSE OF REVIEW: The pathogenesis of DILI is currently unknown; however, research has shown strong genetic associations with some DILIs. This paper describes the variant alleles uncovered by GWAS and discusses their potential role as susceptibility biomarkers. RECENT FINDINGS: An association with HLADRB1*15:01 and amoxicillin/clavulanate DILI has been shown by a number of research groups. The presence of the HLA-B*57:01 allele has been associated with an 81-fold increased risk of flucloxacillin DILI. The HLA-B*35:02 allele has significant association with minocycline DILI. SUMMARY: With the exception of abacavir for HIV therapy, no other prospective genetic screening tests have met the threshold for clinical application. This is largely because DILI incidence is too low to warrant the cost and effort associated with testing. Perhaps, with the development of personalised medicine, a panel of genes for disease susceptibility, drug efficacy and adverse reactions could be tested once off. This would change the cost-effectiveness paradigm, personalise healthcare and reduce DILI risk by avoiding medications in patients with specific HLA alleles.

Systematic review of genetic association studies involving histologically confirmed non-alcoholic fatty liver disease.

UNLABELLED: Non-alcoholic fatty liver disease has an increasing prevalence in Western countries, affecting up to 20% of the population. OBJECTIVE: The aim of this project was to systematically review and summarise the genetic association studies that investigate possible genetic influences that confer susceptibility to non-alcoholic fatty liver disease and non-alcoholic steatohepatitis. DESIGN: The MEDLINE and SCOPUS databases were searched to identify candidate gene studies on histologically diagnosed non-alcoholic fatty liver disease. RESULTS: A total of 85 articles have been summarised and categorised on the basis of the general pathway each candidate gene is involved in, including lipid metabolism, lipoprotein processing, cholesterol synthesis, glucose homoeostasis, inflammatory response, protection against oxidative stress and whole body metabolism. CONCLUSIONS: The main findings demonstrate a small but consistent association of PNPLA3 with non-alcoholic fatty liver disease and non-alcoholic steatohepatitis. Genetic association studies have investigated general disease susceptibility, histological characteristics, severity and progression. However, further study is required to better elucidate the genetic factors influencing fatty liver disease.

Complications and outcomes of phacoemulsification cataract surgery complicated by anterior capsule tear.

PURPOSE: To study the results of phacoemulsification cataract surgery complicated by anterior capsule tear. DESIGN: Retrospective interventional controlled case series. METHODS: Consecutive series of eyes suffering intraoperative anterior capsule tear and others with uneventful cataract surgery at Moorfields Eye Hospital were investigated. Biometric, intraoperative, and postoperative details were recorded. The exclusion criteria were combined surgical procedures, planned manual extracapsular cataract extraction, and history of previous intraocular surgery or eye trauma. The main outcome measures were intraoperative capsule complication rates, refractive and visual outcomes, and incidence of short-term postoperative complications. Two-sided Fisher exact and paired t tests were used for categorical and continuous data, respectively. RESULTS: The study and control groups included 239 and 212 eyes, respectively. In the study group, planned phacoemulsification was converted to manual extraction in 5 cases (2%); a concurrent posterior capsule rupture occurred in 58 eyes (24%) with a rate of nuclear lens material drop in the posterior segment of 5% (11 cases). Over 11% of eyes (n = 27) underwent unplanned secondary surgical procedures. Overall, the refractive outcomes were poor in 22.4% of eyes (postoperative refraction ≥1 diopter of target), and were statistically worse (P < .0001). A significant visual improvement was observed in the majority of the study group eyes (71%); permanent visual loss occurred in 4 eyes (1.7%). CONCLUSIONS: Anterior capsule tear can lead to additional intraoperative complications, with a relatively high incidence of secondary interventions. Overall, permanent visual loss can be observed and worse refractive outcomes are to be expected, particularly if the lens is being implanted out of the bag.

Early diagnosis improves outcomes in hepatitis C.

Chronic hepatitis C (HCV) infection affects 0.8-1.0% of the UK population, with up to 70% having ongoing chronic infection. HCV is curable but if left untreated can progress to end stage liver disease and potentially hepatocellular carcinoma. HCV management options have changed dramatically over the past five years, with improvement in cure rates and tolerability; cure rates of more than 90% can now be achieved. The main risk factors for acquiring HCV infection in the UK are injecting drug use and sharing drug using equipment. Other risk factors include receipt of blood products in the UK before 1991; tattooing or acupuncture with non-sterile equipment; medical procedures; needlestick injuries and contact with blood from an infected person. Acute hepatitis C infection has mild symptoms only and is likely to go undiagnosed. The estimated diagnosis rate in England is 35%, suggesting that 65% of the total HCV-positive population remains undiagnosed. The most common method of detecting HCV is case finding in high- risk groups. Those who test positive for HCV antibodies should be tested for persisting viral presence through HCV PCR testing - a positive result confirms active infection. GPs can play a major role in identifying those at risk of the disease, which includes patients with known risk factors and those with unexplained abnormal liver function tests, providing information and arranging testing. Patients with confirmed active HCV infection should be referred to the local specialist hepatology or infectious disease service in accordance with locally agreed pathways.

Update on the treatment of hepatitis C genotypes 2-6.

PURPOSE OF REVIEW: The hepatitis C virus remains a global health issue, and the established standard of care has consisted of pegylated interferon alpha in conjunction with ribavirin. However, this regimen is associated with significant side-effects and poor sustained virological responses. The aim of this review is to assess the effects of the direct-acting antivirals upon hepatitis C genotypes 2-6 from publications from the past 18 months. RECENT FINDINGS: The impact of direct-acting antivirals has already substantially improved treatments for genotypes 2-6, with the size of improvement much less marked for genotype 3. Although still responsive to these agents, genotype 3 has inherent resistance to treatments possibly owing to its effects on host metabolic pathways. These treatments have moved sustained virological responses to the threshold of 90%, with reduced side-effects and shortened courses of treatment and some options for interferon-free therapy. These newer medications are transforming clinical guidelines at a rapid rate, but this will have to be balanced with the impact it places on global health budgets. SUMMARY: Although direct-acting antivirals are transforming the treatment of all hepatitis C genotypes, ongoing studies will optimize treatment duration and provide interferon-free alternatives.

Serum proteome of nonalcoholic fatty liver disease: a multimodal approach to discovery of biomarkers of nonalcoholic steatohepatitis.

BACKGROUND AND AIM: Nonalcoholic fatty liver disease (NAFLD) is a common condition affecting up to 25% of the developed world. It is a progressive disease, leading in some to the development of liver cirrhosis. Currently, accurate diagnosis and staging of this condition is only possible with histological examination of a liver biopsy. This gold standard test is neither suitable nor practical for large-scale use as is necessary for a condition as common as NAFLD. The aim of this study is to describe the proteome of human NAFLD using two distinct shotgun proteomic methods, translating the findings into potential biomarkers of NAFLD. METHODS: Two distinct shotgun proteomic techniques (iTRAQ and label free) were used to describe the proteome of NAFLD. Thereafter, candidate biomarkers were selected for validation by ELISA. RESULTS: Over 550 protein identifications were made in the description of the NAFLD proteome. Several proteins were found to be significantly up/downregulated in nonalcoholic steatohepatitis compared with control, including apolipoprotein E (fold ratio of 1.67), insulin-like growth factor-binding protein 3 (IGFBP3, fold ratio of 1.642), Vitamin D-binding protein (fold ratio of 4.587), and lymphocyte cytosolic protein1 (LCP1, fold ratio of 4.356). ELISA validation of a subset of these proteins confirms the validity of the proteomic studies and suggests possible new biomarkers of NAFLD. CONCLUSION: Serum markers are able to distinguish between the stages of disease in NAFLD as well as detect the grade of fibrosis. Ultimately, noninvasive serum markers may replace liver biopsy in the investigation of patients with suspected NAFLD.

Bacterial translocation in cirrhosis is not caused by an abnormal small bowel gut microbiota.

Sepsis is common in liver cirrhosis, and animal studies have shown the gut to be the principal source of infection, through bacterial overgrowth and translocation in the small bowel. A total of 33 patients were recruited into this study, 10 without cirrhosis and 23 with cirrhotic liver disease. Six distal duodenal biopsies were obtained and snap frozen for RNA and DNA extraction, or frozen for FISH. Peripheral venous bloods were obtained from 30 patients, including 17 chronic liver disease patients. Samples were analysed by real-time PCR, to assess total bacteria, bifidobacteria, bacteroides, enterobacteria, staphylococci, streptococci, lactobacilli, enterococci, Helicobacter pylori and moraxella, as well as TNF-α, IL-8 and IL-18. There was no evidence of bacterial overgrowth with respect to any of the individual bacterial groups, with the exception of enterococci, which were present in higher numbers in cirrhotic patients (P = 0.04). There were no significant differences in any of the cytokines compared to the controls. The small intestinal mucosal microbiota in cirrhotic patients was qualitatively and quantitatively normal, and this shifts the focus of disease aetiology to factors that reduce gut integrity, failure of mechanisms to remove translocating bacteria, or the large bowel as the source of sepsis.

Systematic review of performance of non-invasive biomarkers in the evaluation of non-alcoholic fatty liver disease.

This systematic review evaluates the many studies carried out to discover and evaluate non-invasive markers of non-alcoholic fatty liver disease (NAFLD). Many different strategies and methods have been used in this task, from the discovery of new markers by global 'shotgun' studies to hypothesis-driven approaches, to the development of algorithm tests based on routinely available clinical and biochemical parameters. We examined the various different approaches, summarising the findings in an attempt to give an overview of the field of non-invasive markers in NAFLD, encompassing markers of steatosis, necro-inflammation and fibrosis. The body of literature surrounding this topic is complex and varied, encompassing not only different methodologies but also different patient characteristics, different disease definitions, as well as different end points. This reflects the heterogeneity of NAFLD, which, however, introduces considerably difficulty when trying to draw a conclusion between studies. We have divided this review into three main chapters based on the characteristics of the studies. The Genomics/Proteomics chapter reviews studies using a non-hypothesis-driven approach to biomarker discovery. Thereafter, we evaluate studies of association - studies that target-specific markers, comparing levels between disease and control groups. Finally, we examine the algorithm tests - mathematical systems developed on the basis of previously described markers and assessed, usually, by receiver operator curve analysis. While radiological examination and investigations offer important diagnostic information, such studies are not discussed in this review - the body of literature surrounding blood and anthropological markers is complex and varied, demanding close attention.

YAG laser peripheral iridotomy for the prevention of pigment dispersion glaucoma a prospective, randomized, controlled trial.

PURPOSE: To test the hypothesis that neodymium:yttrium-aluminum-garnet (Nd:YAG) laser peripheral iridotomy (LPI) significantly reduces the incidence of conversion from pigment dispersion syndrome (PDS) with ocular hypertension (OHT) to pigmentary glaucoma (PG). DESIGN: Prospective, randomized, controlled 3-year trial. PARTICIPANTS: One hundred sixteen eyes of 116 patients with PDS and OHT. INTERVENTION: Patients were assigned randomly either to Nd:YAG LPI or to a control group (no laser). MAIN OUTCOME MEASURES: The primary outcome measure was conversion to PG within 3 years, based on full-threshold visual field (VF) analysis using the Ocular Hypertension Treatment Study criteria. Secondary outcome measures were whether eyes required topical antiglaucoma medications during the study period and the time to conversion or medication. RESULTS: Fifty-seven patients were randomized to undergo laser treatment and 59 were randomized to no laser (controls). Age, gender, spherical equivalent refraction, and intraocular pressure at baseline were similar between groups. Outcome data were available for 105 (90%) of recruited subjects, 52 in the laser treatment group and 53 in the no laser treatment group. Patients were followed up for a median of 35.9 months (range, 10-36 months) in the laser arm and 35.9 months (range, 1-36 months) in the control arm. Eight eyes (15%) in the laser group and 3 eyes (6%) in the control group converted to glaucoma in the study period. The proportion of eyes started on medical treatment was similar in the 2 groups: 8 eyes (15%) in the laser group and 9 eyes (17%) in the control group. Survival analyses showed no evidence of any difference in time to VF progression or commencement of topical therapy between the 2 groups. Cataract extraction was performed on 1 patient in the laser group and in 1 patient in the control group during the study period (laser eye at 18 months; control eye at 34 months). CONCLUSIONS: This study suggests that there was no benefit of Nd:YAG LPI in preventing progression from PDS with OHT to PG within 3 years of follow-up.

Quinolone efflux pumps play a central role in emergence of fluoroquinolone resistance in Streptococcus pneumoniae.

The preferential use of older antimicrobial agents is, in general, sound public health policy and is meant to maintain susceptibility to newer agents. In the case of fluoroquinolones, however, this strategy is flawed and may actually hasten the spread of Streptococcus pneumoniae strains resistant to newer members of the class. In a mouse thigh infection model, we were unable to isolate clones of pneumococci resistant to the newer fluoroquinolone levofloxacin at 2 x or 4 x the baseline MIC. An initial exposure in vivo to the older agent, ciprofloxacin, allowed straightforward selection of clones resistant to levofloxacin in a subsequent experiment. The original ciprofloxacin exposure generated clones without changes in the parC/E and gyrA/B quinolone target sites almost exclusively but did allow overexpression of a reserpine-responsive pump. While this caused only minimal change in the levofloxacin MIC (0.6 mg/liter to 0.8 mg/liter), it allowed a major change in the mutational frequency to resistance for levofloxacin (<1/10(8.5) to approximately 1/10(4.5)), which allowed levofloxacin-resistant clones to be isolated in a subsequent in vivo experiment. The reason underlying ciprofloxacin's propensity to select for pump-overexpressed clones is likely related to its hydrophilicity. To preserve the susceptibility of Streptococcus pneumoniae to newer members of the class of quinolones, use of ciprofloxacin for community-acquired respiratory infections should be minimized.

Cognitive Testing Battery: differences among groups with dementia, Huntington's disease, and controls.

This study evaluated a new test for dementia, the Cognitive Testing Battery, used for over six years in outpatient clinics for aging-related disorders. This battery was developed to be brief, easy to administer, and to provide useful feedback as a cognitive profile for clinicians, patients, and caregivers. 489 participants were tested during a 5-yr. period. Multidisicplinary teams diagnosed these patients with dementia, depression, or Huntington's Disease. A control group was also included. Groups were then compared on the 10 key subscales that subserve dementia populations. Analysis showed there were significant differences among the groups on all measures and that selected scales differentiated the groups. The total score maximally differentiated the groups, and the dementia group was most different from the other groups.

Application of a mathematical model to prevent in vivo amplification of antibiotic-resistant bacterial populations during therapy.

The worldwide increase in the prevalence of multi-antibiotic-resistant bacteria has threatened the physician's ability to provide appropriate therapy for infections. The relationship between antimicrobial drug concentration and infecting pathogen population reduction is of primary interest. Using data derived from mice infected with the bacterium Pseudomonas aeruginosa and treated with a fluoroquinolone antibiotic, a mathematical model was developed that described relationships between antimicrobial drug exposures and changes in drug-susceptible and -resistant bacterial subpopulations at an infection site. Dosing regimens and consequent drug exposures that amplify or suppress the emergence of resistant bacterial subpopulations were identified and prospectively validated. Resistant clones selected in vivo by suboptimal regimens were characterized. No mutations were identified in the quinolone resistance-determining regions of gyrA/B or parC/E. However, all resistant clones demonstrated efflux pump overexpression. At base line, MexAB-OprM, MexCD-OprJ, and MexEF-OprN were represented in the drug-resistant population. After 28 hours of therapy, MexCD-OprJ became the predominant pump expressed in the resistant clones. The likelihood of achieving resistance-suppression exposure in humans with a clinically prescribed antibiotic dose was determined. The methods developed in this study provide insight regarding how mathematical models can be used to identify rational dosing regimens that suppress the amplification of the resistant mutant population.