Increased case-finding and uptake of direct-acting antiviral treatment essential for micro-elimination of hepatitis C among people living with HIV: a national record linkage study.

OBJECTIVES: Micro-elimination of hepatitis C virus (HCV) in people living with HIV (PLHIV) and co-infected with HCV has been proposed as a key contribution to the overall goal of HCV elimination. While other studies have examined micro-elimination in HIV-treated cohorts, few have considered HCV micro-elimination among those not treated for HIV or at a national level. METHODS: Through data linkage of national and sentinel surveillance data, we examined the extent of HCV testing, diagnosis and treatment among a cohort of PLHIV in Scotland identified through the national database of HIV-diagnosed individuals, up to the end of 2017. RESULTS: Of 5018 PLHIV, an estimated 797 (15%) had never been tested for HCV and 70 (9%) of these had undiagnosed chronic HCV. The odds of never having been tested for HCV were the highest in those not on HIV treatment [adjusted odds ratio (aOR) = 7.21, 95% confidence interval (CI): 5.15-10.10). Overall HCV antibody positivity was 11%, and it was at its highest among people who inject drugs (49%). Most of those with chronic HCV (91%) had attended an HCV treatment clinic but only half had been successfully treated (54% for those on HIV treatment, 12% for those not) by the end of 2017. The odds of never having been treated for HCV were the highest in those not on HIV treatment (aOR = 3.60, 95% CI: 1.59-8.15). CONCLUSIONS: Our data demonstrate that micro-elimination of HCV in PLHIV is achievable but progress will require increased effort to engage and treat those co-infected, including those not being treated for their HIV.

A matched comparison study of hepatitis C treatment outcomes in the prison and community setting, and an analysis of the impact of prison release or transfer during therapy.

Prisoners are a priority group for hepatitis C (HCV) treatment. Although treatment durations will become shorter using directly acting antivirals (DAAs), nearly half of prison sentences in Scotland are too short to allow completion of DAA therapy prior to release. The purpose of this study was to compare treatment outcomes between prison- and community-based patients and to examine the impact of prison release or transfer during therapy. A national database was used to compare treatment outcomes between prison treatment initiates and a matched community sample. Additional data were collected to investigate the impact of release or transfer on treatment outcomes. Treatment-naïve patients infected with genotype 1/2/3/4 and treated between 2009 and 2012 were eligible for inclusion. 291 prison initiates were matched with 1137 community initiates: SVRs were 61% (95% CI 55%-66%) and 63% (95% CI 60%-66%), respectively. Odds of achieving a SVR were not significantly associated with prisoner status (P=.33). SVRs were 74% (95% CI 65%-81%), 59% (95% CI 42%-75%) and 45% (95% CI 29%-62%) among those not released or transferred, transferred during treatment, or released during treatment, respectively. Odds of achieving a SVR were significantly associated with release (P<.01), but not transfer (P=.18). Prison-based HCV treatment achieves similar outcomes to community-based treatment, with those not released or transferred during treatment doing particularly well. Transfer or release during therapy should be avoided whenever possible, using anticipatory planning and medical holds where appropriate.

Attendance at specialist hepatitis clinics and initiation of antiviral treatment among persons chronically infected with hepatitis C: examining the early impact of Scotland's Hepatitis C Action Plan.

Primary goals of the Hepatitis C Action Plan for Scotland Phase II (May 2008-March 2011) were to increase, among persons chronically infected with the hepatitis C (HCV) virus, attendance at specialist outpatient clinics and initiation on antiviral therapy. We evaluated progress towards these goals by comparing the odds, across time, of (a) first clinic attendance within 12 months of HCV diagnosis (n = 9747) and (b) initiation on antiviral treatment within 12 months of first attendance (n = 5736). Record linkage between the national HCV diagnosis (1996-2009) and HCV clinical (1996-2010) databases and logistic regression analyses were conducted for both outcomes. For outcome (a), 32% and 45% in the respective pre-Phase II (before 1 May 2008) and Phase II periods attended a specialist clinic within 12 months of diagnosis; the odds of attendance within 12 months increased over time (OR = 1.05 per year, 95% CI: 1.04-1.07), but was not significantly greater for persons diagnosed with HCV in the Phase II era, compared with the pre-Phase II era (OR = 1.1, 95% CI: 0.9-1.3), after adjustment for temporal trend. For outcome (b), 13% and 28% were initiated on treatment within 12 months of their first clinic attendance in the pre-Phase II and Phase II periods, respectively. Higher odds of treatment initiation were associated with first clinic attendance in the Phase II (OR = 1.9, 95% CI: 1.5-2.4), compared with the pre-Phase II era. Results were consistent with a positive impact of the Hepatitis C Action Plan on the treatment of chronically infected individuals, but further monitoring is required to confirm a sustained effect.

Risk factors for postoperative delirium: An umbrella review of systematic reviews.

BACKGROUND: Postoperative delirium (POD) is associated with increased mortality, increased length of hospital stays and increased rates and severity of subsequent cognitive decline including dementia. A wide range of risk factors for POD have been suggested in the literature across multiple surgical specialities. However few are validated and no accurate prognostic models exist. We therefore aimed to map the existing evidence regarding risk factors for POD to help guide future research by undertaking an umbrella review of systematic reviews examining risk factors for POD in any context. MATERIALS AND METHODS: We systematically searched multiple medical databases for systematic reviews examining the risk factors for POD in adults undergoing any surgery. We then selected relevant reviews with minimal overlap in primary studies and extracted information about individual risk factors. RESULTS: Thirty-five relevant reviews were identified of which ten were in trauma and orthopaedic surgery patients (four exclusively examined hip fractures), five were in cardiac surgery patients, and four were in vascular surgery patients. Due to substantial overlap in reviews, eighteen reviews were analysed in detail finding the widely examined and consistent risk factors were increasing age, nursing home residency, pre-existing cognitive impairment, psychiatric disorders, cerebrovascular disease, end stage renal failure, low albumin, higher ASA score, and intra-operative blood transfusion. Many other risk factors were examined, but they were either not studied in multiple systematic reviews, or inconsistent either in results or in categorisation (which for many factors was heterogenous even within systematic reviews). There are also a large number of existing prognostic models, many of which remain unvalidated. CONCLUSION: Given the wealth of existing literature, future research should avoid simple risk factor evaluation except for novel candidates, validate existing prognostic models where possible, and instead focus on interventional research.

Elevation of the provitamin A content of transgenic tomato plants.

Tomato products are the principal dietary sources of lycopene and major source of beta-carotene, both of which have been shown to benefit human health. To enhance the carotenoid content and profile of tomato fruit, we have produced transgenic lines containing a bacterial carotenoid gene (crtI) encoding the enzyme phytoene desaturase, which converts phytoene into lycopene. Expression of this gene in transgenic tomatoes did not elevate total carotenoid levels. However, the beta-carotene content increased about threefold, up to 45% of the total carotenoid content. Endogenous carotenoid genes were concurrently upregulated, except for phytoene synthase, which was repressed. The alteration in carotenoid content of these plants did not affect growth and development. Levels of noncarotenoid isoprenoids were unchanged in the transformants. The phenotype has been found to be stable and reproducible over at least four generations.

The purification of phytoene dehydrogenase from Phycomyces blakesleeanus.

The carotenogenic enzyme phytoene dehydrogenase has been purified from the C9carR21(-) (lycopene-accumulating) mutant of the filamentous fungus Phycomyces blakesleeanus. Solubilization of the membrane-bound enzyme with 1% Tween-60 was followed by a 250-fold purification to homogeneity using polyethylene glycol precipitation, CM-Sepharose, gel filtration and isoelectric focusing. Multiple peaks of enzymic activity were found in eluates from ion-exchange and gel filtration chromatography, with the lowest molecular weight fraction having an apparent molecular mass of approx. 14 kDa. All active fractions catalyzed the dehydrogenation of 15-cis phytoene into all-trans lycopene, with a cis-trans isomerization occurring at phytofluene. Both NADP+ and FAD were required for the dehydrogenation reaction. The presence of > 0.5% Tween-60 was necessary to maintain enzymic activity, although in its absence lipids restored some activity. The enzyme could be stored for at least 6 weeks at -70 degrees C in the presence of 20% (v/v) glycerol.

The subcellular distribution of carotenoids in Phycomyces blakesleeanus C115 car-42 mad-107 (--).

1. Subcellular fractions of the C115 car-42 mad-107(--) strain of Phycomyces blakesleeanus, prepared by differential centrifugation, were irreversibly associated with extraneous lipids. Such contamination was avoided by the use of density gradient centrifugation. 2. The two beta-carotene-containing fractions, a particulate fraction and lipid globules, also contained sterol, phospholipid and alkaline p-nitrophenylphosphatase activity. Both fractions revealed close similarities on amino acid analysis.

Characterization of ent-kaurene oxidase activity from Gibberella fujikuroi.

Cell extracts of wild-type and mutant strains of Gibberella fujikuroi were assayed for kaurene oxidase activity, using ent-[3H]kaurene as the substrate. Extracts from strain SG78 exhibited the highest specific activity, and were used in subsequent experiments. The microsomal enzyme activity was solubilized with buffers or salt solutions at a concentration of 400 mM. Both the soluble and microsomal preparations showed characteristic cytochrome P-450 spectra, ligand binding spectra with the substrate and with the plant growth regulator, paclobutrazol, and inhibition of enzymic activity by carbon monoxide. The addition of 20% glycerol to the extraction buffer stabilized the activity to some extent. Loss of enzymic activity on storage was accompanied by conversion of P-450 to P-420. Michaelis-Menten kinetic parameters for the membrane-bound and soluble enzyme have been estimated, as have constants for the binding of ent-kaurene and paclobutrazol to the P-450 and P-420 forms of the protein.

Carotenoid biosynthesis in wild type and mutant strains of Mucor circinelloides.

Carotenoid biosynthesis in wild type Mucor circinelloides has been investigated and the biochemical characterisation of the MS1 and MS9 mutant strains, impaired in carotenoid formation, carried out. In liquid cultures, all strains produced carotenoids (mainly beta-carotene, but also xi-carotene, lycopene and gamma-carotene) at the onset of stationary phase of growth. Carotenogenesis was light dependent. In liquid cultures carotenoid formation in wild type was affected by diphenylamine, which prevented desaturation, nicotine, resulting in reduced carotenoid levels, but CPTA caused an increase in the total carotenoid content but a reduced beta-carotene level, with the accumulation of lycopene and gamma-carotene. The mutant strains MS1 and MS9 contained only 5.0 and 11.5% of wild type carotenoid levels, respectively. Cell extracts of light-grown mycelia, incubated with 3(R)-[2-14C] mevalonic acid, produced beta-carotene, but incorporations into carotenoids were substantially reduced in the cell extracts of MS1 and MS9. Analysis of prenyl diphosphate intermediates indicated that, compared to wild type, geranylgeranyl diphosphate accumulated in MS1. MS9 extracts produced a larger amount of prenyl phosphates and a more even distribution of radioactivity from mevalonic acid into farnesyl and geranylgeranyl diphosphates. Squalene and long chain prenyl phosphates were formed by the cell extracts of all strains. It is proposed that the MS1 strain possesses a mutation in a gene responsible for phytoene formation, whilst a regulatory mutation, affecting prenyl transferase activities has occurred in MS9.

Carotenoid Biosynthesis during Tomato Fruit Development (Evidence for Tissue-Specific Gene Expression).

Tomato (Lycopersicon esculentum Mill. cv Ailsa Craig) fruit, at five stages of development, have been analyzed for their carotenoid and chlorophyll (Chl) contents, in vitro activities of phytoene synthase, phytoene desaturase, and lycopene cyclase, as well as expression of the phytoene synthase (Psy) and phytoene desaturase (Pds) genes. During ripening, the total carotenoids increased with a concomitant decrease in Chl. Although the highest carotenoid content (consisting mainly of lycopene and [beta]-carotene) was found in ripe fruit, the greatest carotenogenic enzymic activities were found in green fruit. Phytoene synthase was located in the plastid stroma, whereas the metabolism of phytoene was associated with plastid membranes during all stages of fruit development. The in vitro products of phytoene desaturation altered from being predominantly phytofluence and [zeta]-carotene in chloroplasts to becoming mainly lycopene in chromoplasts. The expression of Psy was detected in breaker and ripe fruit, as well as flowers, but was not detectable by northern blot analysis in leaves or green fruits. The Pds gene transcript was barely detectable in green fruit and leaves but was expressed in flowers and breaker fruit. These results suggest that transcription of Psy and Pds is regulated developmentally, with expression being considerably elevated in chromoplast-containing tissues. Antiserum to the Synechococcus phytoene synthase cross-reacted with phytoene synthase of green fruit only on western blots and not with the enzyme from ripe fruit. In contrast, a monoclonal antibody to the Psy gene product only cross-reacted with phytoene synthase from ripe fruit. The enzymes from green and ripe fruit had different molecular masses of 42 and 38 kD, respectively. The absence of detectable Psy and Pds mRNA in green tissues using northern blot analyses, despite high levels of phytoene synthase and desaturase activity, lends support to the hypothesis of divergent genes encoding these enzymes.

Expression of prokaryotic 1-deoxy-D-xylulose-5-phosphatases in Escherichia coli increases carotenoid and ubiquinone biosynthesis.

Isopentenyl diphosphate (IPP) acts as the common, five-carbon building block in the biosynthesis of all isoprenoids. The first reaction of IPP biosynthesis in Escherichia coli is the formation of 1-deoxy-D-xylulose-5-phosphate, catalysed by 1-deoxy-D-xylulose-5-phosphate synthase (DXPS). E. coli engineered to produce lycopene, was transformed with dxps genes cloned from Bacillus subtilis and Synechocystis sp. 6803. Increases in lycopene levels were observed in strains expressing exogenous DXPS compared to controls. The recombinant strains also exhibited elevated levels of ubiquinone-8. These increases corresponded with enhanced DXP synthase activity in the recombinant E. coli strains.

Antioxidant activities of carotenes and xanthophylls.

The purpose of this study was to assess the relative antioxidant activities of a range of carotenes and xanthophylls through the extent of their abilities to scavenge the ABTS(.+) radical cation. The results show that the relative abilities of the carotenoids to scavenge the ABTS(.+) radical cation are influenced by the presence of functional groups with increasing polarities, such as carbonyl and hydroxyl groups, in the terminal rings, as well as by the number of conjugated double bonds.

Intracellular cations and transmembrane cation transport in essential hypertension: the importance of controlled clinical observations.

We measured Na+, K+ and total Ca2+ concentrations as well as first order efflux rate constants for Na+ and Ca2+ in red cells from 22 patients with untreated essential hypertension and 22 normotensive control subjects. Both groups were individually and prospectively matched for age, sex and weight. Intracellular Na+ and K+ were also measured in a separate population consisting of 31 hypertensive patients and 41 normotensive subjects who were not matched. In order to test for possible plasma inhibitors of Na+ or Ca2+ transport in hypertension, Na+ and Ca2+ efflux rate constants in 15 paired patients were measured in red cells which were incubated in either homologous or paired plasma. Intracellular Na+ and K+ concentrations and Na+ and Ca2+ efflux rate constants were similar in the matched subjects. Significant positive correlations were found between intracellular Na+ and body weight (r = 0.27, P = 0.041) and between intracellular K+ and age (r = 0.29, P = 0.028). In the unmatched population intracellular Na+ correlated significantly with age (r = 0.25, P = 0.029) and was significantly increased in hypertensive patients. This difference was due wholly to increased intracellular Na+ in hypertensive females. After correction for age and weight, intracellular Na+ was similar in both hypertensive and normotensive subjects. Intracellular K+ did not differ between these groups but was significantly lower in males. Intracellular Ca2+ was similar in the matched groups after correction for age and weight, but the regression equations containing age and weight as independent variables were significantly different in hypertensive and normotensive subjects.(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of a tomato cDNA coding for phytoene synthase in Escherichia coli, phytoene formation in vivo and in vitro, and functional analysis of the various truncated gene products.

Full length and truncated cDNA expression constructs of the phytoene synthase (psy) gene from tomato have been ligated into a pUC8 cloning vector. One of the truncated constructs was introduced into Escherichia coli carrying the Erwinia uredovora GGPP synthase gene. This transformant produced 15,15'-cis-phytoene, which was identified on the basis of its UV and IR spectral data, from geranylgeranyl diphosphate. The function of this gene product was further confirmed by in vitro assay using cell-free extract of E. coli harboring the construct. On transformation with the above constructs together with a plasmid containing the carotenoid gene cluster from E. uredovora devoid of the phytoene synthase (crtB) gene, yellow, carotenoid-containing, E. coli colonies were produced. The amounts of carotenoids synthesized by the transformed cells, related to the steady-state levels of psy mRNA, varied depending upon the psy constructs. The full-length psy clone produced 16-fold less carotenoids per unit amount of RNA than cells containing phytoene synthase without the first 114 N-terminal amino acids. Removal of further amino acids from the N-terminus caused a large decrease in carotenogenesis. A Western blot of ripe fruit stroma with a monoclonal antibody raised against phytoene synthase revealed a single protein band of apparent molecular mass 38 kDa. Based upon this immunological evidence, we conclude that the size of the transit peptide of phytoene synthase from ripe tomato fruit is approximately 9 kDa, corresponding to about 80 amino acid residues.(ABSTRACT TRUNCATED AT 250 WORDS)

Yeast flora of the mouth and skin during and after irradiation for oral and laryngeal cancer.

The quantitative and qualitative changes occurring the fungal flora of 22 patients with oral and 9 with laryngeal carcinoma were studied during and after radiation therapy. Each patient received 6000 rad of externally applied radiation in divided doses for 5 weeks. The fungal flora was isolated from the patients' oral cavity and irradiated skin sites during irradiation and 2 weeks and 4-6 months afterwards. The number and types of fungi increased in both groups of patients after the start of irradiation and persisted at high levels for at least 4-6 months after treatment. Candida albicans and C. tropicalis were the principal yeasts isolated throughout the period studied but seven other species were also identified. All the yeast isolates were sensitive in vitro to miconazole, ketoconazole, amphotericin B and nystatin. Any of these antifungal agents should be appropriate for therapy.

The relative antioxidant activities of plant-derived polyphenolic flavonoids.

The relative antioxidant activities, against radicals generated in the aqueous phase, of a range of plant-derived polyphenolic flavonoids, constituents of fruit, vegetables, tea and wine, have been assessed. The results show that compounds such as quercetin and cyanidin, with 3',4' dihydroxy substituents in the B ring and conjugation between the A and B rings, have antioxidant potentials four times that of Trolox, the vitamin E analogue. Removing the ortho-dihydroxy substitution, as in kaempferol, or the potential for electron delocalisation by reducing the 2,3 double bond in the C ring, as in catechin and epicatechin, decreases the antioxidant activity by more than 50%, but these structures are still more effective than alpha-tocopherol or ascorbate. The relative significance of the positions and extents of hydroxylation of the A and B rings to the total antioxidant activity of these plant polyphenolics is demonstrated.

Isomerization of dietary lycopene during assimilation and transport in plasma.

Diets of individuals were supplemented with tomatoes, either cooked or as tomato pureé in order to compare uptake of lycopene from intact and homogenized fruit tissue matrices. Following a diet containing cooked tomatoes over three consecutive 7-day periods, little change in the carotenoid levels in plasma lipoproteins occurred. In contrast, a diet supplemented with concentrated tomato pureé, over a 2 week period, caused a significant (p < 0.05) increase in lycopene levels in plasma, showing that the lycopene within intact cells is less bioavailable than that from processed tissue. The isomeric composition of plasma lycopene was significantly different to that of the ingested pureé. A number of cis-isomers (predominantly 5-cis, 13-cis and 9-cis-) were detected in plasma, that are not present in the lycopene from pureé. The significance of the increase in lycopene following dietary supplementation with respect to bioavailability and the causes of isomerization are discussed.