Quantitative Assessment of Secondary Healthcare Utilisation by Patients With Functional Abdominal Pain.

There is increasing awareness of the impact functional conditions have on the National Health Service (NHS). Less is known about the resources used to manage these conditions. This retrospective quantitative audit aims to determine the demographic and healthcare utilisation of functional abdominal pain patients presenting to the hospital. The most frequent hospital attenders with non-specific abdominal pain in NHS Grampian, 2018-2019, were assessed (n=144). Adult patients meeting the ROME II diagnostic criteria for functional abdominal pain diagnosis were included (n=33). Data were retrospectively collected manually from electronic medical records. Of 33 patients, 93.9% were female, with a mean age of 31.2 years. Each had accessed a mean of 11.5 specialist services, with 69.7% being referred to mental health services; 9.1% had completed treatment. Each patient had a median 4 (range 1-26) emergency/unscheduled presentations to hospital and median 2 (range 0-13) admissions for functional abdominal pain during the study period, with a total of 247 nights spent in hospital by this patient cohort for functional abdominal pain alone. The estimated total cost for these hospital admissions was £593,786.00. Extensive secondary-care input is currently required for patients with functional abdominal pain at a significant cost. Patients are re-presenting to the hospital frequently, which suggests that current management is not effective.

Multiple sclerosis, emotion perception and social functioning.

People with multiple sclerosis (MS) can experience problems in interpreting others' emotions from faces or voices. However, to date little is known about whether difficulties in emotion perception in MS are related to broader aspects of social functioning. Also, there are few studies reporting the effect of MS on more ecologically valid assessments of emotion perception using multimodal videos. The current study looks at (1) the effect of MS on perceiving emotions from faces, voices and multimodal videos; (2) the possible role of slowed processing and executive dysfunction in emotion perception problems in MS and (3) the relationship between emotion perception and broader social functioning in MS. 53 people with MS and 31 healthy controls completed tasks of emotion perception and cognition, and assessed their levels of social support and social participation. Participants with MS performed worse than demographically matched controls on all measures of emotion perception. Emotion perception performance was related to cognitive measures in those with MS. Also, significant associations were found between emotion perception difficulties in MS and poorer social function. In particular, people with MS who had poorer emotion perception also reported lower levels of social support from their friends, and regression analysis showed that this prediction was maintained even when disease severity and cognitive function were taken into account. These results show that problems with emotion perception in MS extend to more realistic tasks and may predict key aspects of social functioning.

The nature of anger in people with multiple sclerosis: a qualitative study.

Objective: People with multiple sclerosis (pwMS) who experience higher levels of anger also report poorer quality of life (QoL). This qualitative study explored the subjective experience of anger amongst pwMS, and how anger influenced their lives.Methods: A series of semi-structured, face-to-face interviews were conducted with 20 pwMS. Interviews were recorded, transcribed and then Interpretative Phenomenological Analysis was used to analyse the emerging themes.Results: The most common experience of anger was frustration that MS-related symptoms restricted participation in everyday activities. Also, some experiences of anger-with-self were focused on frustration at the inability to overcome symptom-related activity limitations. Participants reported frustration with others' insensitivity to the effects of the disease process, as well as usual daily irritations with family and colleagues. Some of the participants reported the use of coping strategies to deal with anger episodes.Conclusion: Many pwMS experience frustration at the restrictions that the disease places on them, self-directed anger, and irritation with others' attitude towards them. Much research in MS focuses on physical symptoms, but current results indicate that there is a need to better understand the emotional challenges faces by pwMS, and to provide more support for those who are experiencing frustration.

Free radical generation from an aniline derivative in HepG2 cells: a possible captodative effect.

Xenobiotic metabolism can induce the generation of protein radicals, which are believed to play an important role in the toxicity of chemicals and drugs. It is therefore important to identify chemical structures capable of inducing macromolecular free radical formation in living cells. In this study, we evaluated the ability of four structurally related environmental chemicals, aniline, nitrosobenzene, N,N-dimethylaniline, and N,N-dimethyl-4-nitrosoaniline (DMNA), to induce free radicals and cellular damage in the hepatoma cell line HepG2. Cytotoxicity was assessed using lactate dehydrogenase assays, and morphological changes were observed using phase contrast microscopy. Protein free radicals were detected by immuno-spin trapping using in-cell western experiments and confocal microscopy to determine the subcellular locale of free radical generation. DMNA induced free radical generation, lactate dehydrogenase release, and morphological changes in HepG2 cells, whereas aniline, nitrosobenzene, N,N-dimethylaniline did not. Confocal microscopy showed that DMNA induced free radical generation mainly in the cytosol. Preincubation of HepG2 cells with N-acetylcysteine and 2,2'-dipyridyl significantly prevented free radical generation on subsequent incubation with DMNA, whereas preincubation with apocynin and dimethyl sulfoxide had no effect. These results suggest that DMNA is metabolized to reactive free radicals capable of generating protein radicals which may play a critical role in DMNA toxicity. We propose that the captodative effect, the combined action of the electron-releasing dimethylamine substituent, and the electron-withdrawing nitroso substituent, leads to a thermodynamically stabilized radical, facilitating enhanced protein radical formation by DMNA.

Difficulties with emotion regulation in multiple sclerosis: Links to executive function, mood, and quality of life.

INTRODUCTION: Little is known about the influence of multiple sclerosis (MS) on the regulation of emotion. The current study tested whether people with MS report more emotion regulation difficulties than healthy controls. The relationship between emotion regulation and other important variables (mood, quality of life, and executive function) was explored. Mediation models were used to further understand the links between emotion regulation, depressed mood, and executive function in MS. METHOD: A total of 31 people with MS and 31 controls completed the Difficulties in Emotion Regulation Scales and measures of executive function (fluency and a go/no-go task), mood (Hospital Anxiety and Depression Scales), and a multidimensional assessment of quality of life (World Health Organization Quality of Life, brief version). RESULTS: People with MS reported experiencing more difficulties in emotion regulation than controls. Mediation analyses indicated that depression mediated the emotion regulation difficulties in MS, while executive dysfunction did not. Difficulties in emotion regulation predicted poorer psychological and social quality of life in MS, independent of problems with executive function. CONCLUSIONS: People with MS experience difficulties in emotion regulation, which predict poorer quality of life. These results indicate that emotional control skills should be investigated in further detail when considering interventions to enhance well-being in MS.

Investigating free radical generation in HepG2 cells using immuno-spin trapping.

Oxidative stress can induce the generation of free radicals, which are believed to play an important role in both physiological and pathological processes and a number of diseases such as cancer. Therefore, it is important to identify chemicals which are capable of inducing oxidative stress. In this study, we evaluated the ability of four environmental chemicals, aniline, nitrosobenzene (NB), N,N-dimethylaniline (DMA) and N,N-dimethyl-4-nitrosoaniline (DMNA), to induce free radicals and cellular damage in the hepatoma cell line HepG2. Cytotoxicity was assessed using lactate dehydrogenase (LDH) assays and morphological changes were observed using phase contrast microscopy. Free radicals were detected by immuno-spin trapping (IST) in in-cell western experiments or in confocal microscopy experiments to determine the subcellular localization of free radical generation. DMNA induced free radical generation, LDH release and morphological changes in HepG2 cells whereas aniline, NB and DMA did not. Confocal microscopy showed that DMNA induced free radical generation mainly in the cytosol. Preincubation of HepG2 cells with N-acetylcysteine and 2,2'-dipyridyl significantly prevented free radical generation upon subsequent incubation with DMNA, whereas preincubation with apocynin and dimethyl sulfoxide did not. These results suggest that DMNA induces oxidative stress and that reactive oxygen species, metals and free radical generation play a critical role in DMNA-induced cytotoxicity.

The mini-mental Parkinson's (MMP) as a cognitive screening tool in people with Parkinson's disease.

BACKGROUND: Cognitive decline is common in Parkinson's disease (PD) but may not be adequately identified by the mini-mental state examination (MMSE), which is better suited to Alzheimer's disease. The mini-mental Parkinson (MMP) examination is a cognitive screening tool designed in French specifically for PD. We aimed to establish the validity and reliability of the English language version of the MMP compared with the MMSE. METHODS: People with various stages of PD underwent testing with the MMP and MMSE, which was then compared with a reference standard battery of neuropsychological tests to identify those with significant cognitive impairment. RESULTS: Forty-nine patients were recruited. Both the MMP and MMSE were significantly correlated with scores on all the neuropsychological tests in the validation battery. The median MMP score was proportionally lower (80% of maximum) than the MMSE (90% of maximum) in PD patients with cognitive impairment and those with prior neuropsychiatric complications but there was no difference between the MMP and MMSE in areas under the curves (0.84) for detecting cognitive impairment. Test-retest reliability of the MMP was good (intra-class correlation coefficient 0.793). An MMP of 28 or lower out of 32 detected cognitive impairment with 87% sensitivity and 76% specificity. DISCUSSION: The English language version of the MMP has now been validated. It detects more cognitive deficits in PD patients than the MMSE and identifies significant cognitive impairment in those with PD at least as well as the MMSE.

Photooxidation of Amplex Red to resorufin: implications of exposing the Amplex Red assay to light.

The Amplex Red assay, a fluorescent assay for the detection of H2O2, relies on the reaction of H2O2, which, in the presence of horseradish peroxidase, oxidizes the colorless, nonfluorescent, Amplex Red with a 1:1 stoichiometry to form the colored, fluorescent resorufin. We have found that resorufin is artifactually formed when Amplex Red is exposed to light. This photochemistry is initiated by trace amounts of resorufin present in Amplex Red stock solutions. ESR spin-trapping studies have demonstrated that superoxide radical is an intermediate in this process. Oxygen consumption measurements further confirmed that superoxide and H2O2 were artifactually produced by the photooxidation of Amplex Red. The artifactual formation of resorufin was also significantly increased by the presence of superoxide dismutase or HRP. This photooxidation process leads to a less sensitive assay for H2O2 under ambient light exposure and potentially invalid measurements under high energy exposure such as UVA irradiation. In general, precautions should be taken to minimize exposure to light, including that from instrumental light, during measurement of oxidative stress with Amplex Red.

Development of immunoblotting techniques for DNA radical detection.

Radical damage to DNA has been implicated in cell death, cellular dysfunction, and cancer. A recently developed method for detecting DNA radicals uses the nitrone spin trap DMPO (5,5-dimethyl-1-pyrroline N-oxide) to trap radicals. The trapped radicals then decay into stable nitrone adducts detectable with anti-DMPO antibodies and quantifiable by ELISA or dot-blot assay. However, the sequences of DNA that are damaged are likely to be as important as the total level of damage. Therefore, we have developed immunoblotting methods for detection of DNA nitrone adducts on electrophoretically separated DNA, comparable to Western blotting for proteins. These new techniques not only allow the assessment of relative radical adduct levels, but can reveal specific DNA fragments, and ultimately nucleotides, as radical targets. Moreover, we have determined that denaturation of samples into single-stranded DNA enhances the detection of DNA-DMPO adducts in our new blotting methods and also in ELISA.

Site-specific detection of radicals on α-lactalbumin after a riboflavin-sensitized reaction, detected by immuno-spin trapping, ESR, and MS.

Free radicals and other oxidation products were characterized on α-lactalbumin with electron spin resonance (ESR), immuno-spin trapping, and mass spectrometry (MS) after riboflavin-mediated oxidation. Radicals were detected using the spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) in immuno-spin trapping with both enzyme-linked immunosorbent assay (ELISA) and Western blotting and further characterized with mass spectrometry. A DMPO-trapped radical was identified at His68 and another at one of the tyrosine residues, Tyr50 or Tyr36, respectively, generated by a type II or I mechanism. Not all tyrosyl radicals were trapped, as the secondary oxidation product, 3,4-dihydroxyphenylalanine (DOPA), was detected by mass spectrometry at Tyr18 and Tyr50. A further oxidation of DOPA resulted in the DOPA o-semiquinone radical, which was characterized by ESR. Both surface exposure and the neighboring residues in the local environment of the tertiary structure of α-lactalbumin seem to play a role in the generation of DMPO trapped radicals and secondary oxidation products.

Identification of proteins susceptible to thiol oxidation in endothelial cells exposed to hypochlorous acid and N-chloramines.

Hypochlorous acid (HOCl) is a potent oxidant produced by the enzyme myeloperoxidase, which is released by neutrophils under inflammatory conditions. Although important in the immune system, HOCl can also damage host tissue, which contributes to the development of disease. HOCl reacts readily with free amino groups to form N-chloramines, which also cause damage in vivo, owing to the extracellular release of myeloperoxidase and production of HOCl. HOCl and N-chloramines react readily with cellular thiols, which causes dysfunction via enzyme inactivation and modulation of redox signaling processes. In this study, the ability of HOCl and model N-chloramines produced on histamine and ammonia at inflammatory sites, to oxidize specific thiol-containing proteins in human coronary artery endothelial cells was investigated. Using a proteomics approach with the thiol-specific probe, 5-iodoacetamidofluorescein, we show that several proteins including peptidylprolyl isomerase A (cyclophilin A), protein disulfide isomerase, glyceraldehyde-3-phosphate dehydrogenase and galectin-1 are particularly sensitive to oxidation by HOCl and N-chloramines formed at inflammatory sites. This will contribute to cellular dysfunction and may play a role in inflammatory disease pathogenesis.

Photooxidation of Amplex Red to resorufin: implications of exposing the Amplex Red assay to light.

The Amplex Red assay, a fluorescent assay for the detection of H(2)O(2), relies on the reaction of H(2)O(2) and colorless, nonfluorescent Amplex Red with a 1:1 stoichiometry to form colored, fluorescent resorufin, catalyzed by horseradish peroxidase (HRP). We have found that resorufin is artifactually formed when Amplex Red is exposed to light. In the absence of H(2)O(2) and HRP, the absorption and fluorescence spectra of Amplex Red changed during exposure to ambient room light or instrumental excitation light, clearly indicating that the fluorescent product resorufin had formed. This photochemistry was initiated by trace amounts of resorufin that are present in Amplex Red stock solutions. ESR spin-trapping studies demonstrated that superoxide radical was an intermediate in this process. Oxygen consumption measurements further confirmed that superoxide and H(2)O(2) were artifactually produced by the photooxidation of Amplex Red. The artifactual formation of resorufin was also significantly increased by the presence of superoxide dismutase or HRP. This photooxidation process will result in a less sensitive assay for H(2)O(2) under ambient light exposure and potentially invalid measurements under high energy exposure such as UVA irradiation. In general, precautions should be taken to minimize exposure to light during measurement of oxidative stress with Amplex Red.

High plasma thiocyanate levels in smokers are a key determinant of thiol oxidation induced by myeloperoxidase.

Smokers have an elevated risk of atherosclerosis but the origins of this elevated risk are incompletely defined, though evidence supports an accumulation of the oxidant-generating enzyme myeloperoxidase (MPO) in the inflamed artery wall. We hypothesized that smokers would have a high level of thiocyanate (SCN(-)), a preferred substrate for MPO, which in turn would predispose to thiol oxidation, an established independent risk factor for atherosclerosis. In this study it is shown that on exposure to MPO/H(2)O(2), thiols on plasma proteins from nonsmokers were increasingly oxidized with increasing added SCN(-) concentrations. Plasma from smokers contained significantly higher endogenous levels of SCN(-) than that from nonsmokers (131±31 vs 40±24 µM, P<0.0001). When plasma from smokers and nonsmokers was exposed to MPO/H(2)O(2)-stimulated oxidation, a strong positive correlation (r=0.8139, P<0.0001) between the extent of thiol oxidation and the plasma SCN(-) concentrations was observed. Computational calculations indicate a changeover from HOCl to HOSCN as the major MPO-generated oxidant in plasma, with increasing SCN(-) levels. These data indicate that plasma SCN(-) levels are a key determinant of the extent of thiol oxidation on plasma proteins induced by MPO, and implicate HOSCN as an important mediator of inflammation-induced oxidative damage to proteins in smokers.

Specific impairments of emotion perception in multiple sclerosis.

OBJECTIVE: Multiple sclerosis (MS) often results in demyelination of a network of frontal-subcortical tracts involved in processing emotional information. We investigated the effect of MS on the ability to identify emotional and nonemotional information from static and dynamic stimuli and determined whether difficulties in emotion perception related to quality of life. METHOD: 32 MS and 33 control participants, matched for age and education, identified emotions and nonemotional information from static images of faces and dynamic videos of people interacting. They also completed cognitive assessment and quality of life ratings. RESULTS: On the static face perception tasks, participants with MS performed more poorly than healthy controls on emotion perception, t(63) = 3.30, p < .01, d = .83, but not identity perception, t(63) = 1.18, d = .30. For the dynamic tasks, the MS group were impaired on emotion perception, t(63) = 3.41, p = .001, d = .86, but not age/gender perception, t(63) = 0.15, d = .04. Ratings of social and psychological aspects of quality of life in MS were related to emotion perception scores, controlling for disease severity and duration, age, depression, and cognitive function, with r2 ranging from .17 to .24. CONCLUSIONS: These results indicate a specific deficit in decoding static and dynamic information about emotion in MS, as compared to nonemotional information. There were specific relationships between emotion perception problems and poor social and psychological quality of life, indicating that emotional skills should be considered when evaluating functioning in MS.

Protein Radical Formation Resulting from Eosinophil Peroxidase-catalyzed Oxidation of Sulfite.

Eosinophil peroxidase (EPO) is an abundant heme protein in eosinophils that catalyzes the formation of cytotoxic oxidants implicated in asthma, allergic inflammatory disorders, and cancer. It is known that some proteins with peroxidase activity (horseradish peroxidase and prostaglandin hydroperoxidase) can catalyze oxidation of bisulfite (hydrated sulfur dioxide), leading to the formation of sulfur trioxide anion radical ((.)SO(3)(-)). This free radical further reacts with oxygen to form peroxymonosulfate anion radical ((-)O(3)SOO(.)) and the very reactive sulfate anion radical (SO(4)()), which is nearly as strong an oxidant as the hydroxyl radical. However, the ability of EPO to generate reactive sulfur radicals has not yet been reported. Here we demonstrate that eosinophil peroxidase/H(2)O(2) is able to oxidize bisulfite, ultimately forming the sulfate anion radical (SO(4)()), and that these reactive intermediates can oxidize target proteins to protein radicals, thereby initiating protein oxidation. We used immuno-spin trapping and confocal microscopy to study protein oxidation by EPO/H(2)O(2) in the presence of bisulfite in a pure enzymatic system and in human promyelocytic leukemia HL-60 clone 15 cells, maturated to eosinophils. Polyclonal antiserum raised against the spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) detected the presence of DMPO covalently attached to the proteins resulting from the DMPO trapping of protein free radicals. We found that sulfite oxidation mediated by EPO/H(2)O(2) induced the formation of radical-derived DMPO spin-trapped human serum albumin and, to a lesser extent, of DMPO-EPO. These studies suggest that EPO-dependent oxidative damage may play a role in tissue injury in bisulfite-exacerbated eosinophilic inflammatory disorders.

Attentional lapses, emotional regulation and quality of life in multiple sclerosis.

OBJECTIVES: We investigated the role of attentional lapses and emotion regulation in predicting multiple aspects of Quality of Life (QoL) in MS. METHODS: Individuals with MS (n=86) reported on frequency of attentional lapses using the Cognitive Failures Questionnaire, deployment of different emotion regulation strategies using the Emotion Regulation Questionnaire, and QoL on a range of domains using the WHOQoL-BREF. RESULTS: Regression analyses indicated that attentional failures and infrequent use of emotional reappraisal strategies predicted variance in all aspects of QoL, in addition to effects of disease severity (total R(2)s ranged from .134 to .446). CONCLUSIONS: Problems with attentional lapses and a failure to use effective emotion regulation strategies may contribute to reduced QoL in MS.

Identification of plasma proteins that are susceptible to thiol oxidation by hypochlorous acid and N-chloramines.

Hypochlorous acid (HOCl), the major strong oxidant produced by myeloperoxidase, reacts readily with free amino groups to form N-chloramines. Although HOCl and N-chloramines play an important role in the human immune system by killing bacteria and invading pathogens, they have also been shown to cause damage to tissues, which is believed to contribute to a number of diseases. It has been shown previously that N-chloramines react more readily with protein thiols than with other targets in plasma, but the nature of the plasma thiol-containing proteins oxidized is unknown. In this study, the ability of N-chloramines to selectively oxidize thiol-containing plasma proteins was determined using the thiol-specific probe, 5-iodoacetamidofluorescein, combined with two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Experiments were performed with N-chloramines formed on Nalpha-acetyl-lysine, Nalpha-acetyl-histidine (HisCA), glycine, taurine, and ammonia. With the exception of HisCA, the N-chloramines were more efficient than HOCl at oxidizing plasma thiols. The thiol-containing plasma proteins alpha1-antitrypsin and transthyretin were found to be oxidized in addition to albumin, with this treatment resulting in the inactivation of alpha1-antitrypsin. A similar selectivity of reaction and extent of thiol oxidation were also observed with myeloperoxidase in the presence of hydrogen peroxide and chloride ions.

Traumatic brain injury and prospective memory: influence of task complexity.

A quantitative review indicated that prospective memory impairment is a consistent feature of traumatic brain injury (TBI). However, evidence also suggests that manipulations that increase demands on controlled attentional processes moderate the magnitude of observed deficits. A total of 16 TBI participants were compared with 15 matched controls on a task in which the number of prospective target events was manipulated. This manipulation was of interest because two competing models make different predictions as to its effect on controlled attentional processes. In the context of Smith and Bayen's (2004) preparatory attentional processes and memory processes (PAM) model increasing the number of target events should increase requirements for controlled attentional processing. In contrast, McDaniel and Einstein's (2000) multiprocess framework assumes that distinct target events presented in focal awareness of the processing activities required for the ongoing task are likely to depend on automatic processes. This latter model therefore leads to the prediction that increasing the number of target events should not increase demands upon controlled attentional processes. Consistent with McDaniel and Einstein's (2000) multiprocess framework, TBI patients were significantly and comparably impaired on the one- and the four-target-event conditions relative to controls. Further, TBI deficits could not be attributed to increased difficulty with the retrospective component of the prospective memory task. The practical and theoretical implications of these results are discussed.

Theory of mind following traumatic brain injury: the role of emotion recognition and executive dysfunction.

A number of studies have now documented that traumatic brain injury (TBI) is associated with deficits in the recognition of basic emotions, the capacity to infer mental states of others (theory of mind), as well as executive functioning. However, no study to date has investigated the relationship between these three constructs in the context of TBI. In the current study TBI participants (N=16) were compared with demographically matched healthy controls (N=17). It was found that TBI participants' recognition of basic emotions, as well as their capacity for mental state attribution, was significantly reduced relative to controls. Performance on both of these measures was strongly correlated in the healthy control, but not in the TBI sample. In contrast, in the TBI (but not the control) sample, theory of mind was substantially correlated with performance on phonemic fluency, a measure of executive functioning considered to impose particular demands upon cognitive flexibility and self-regulation. These results are consistent with other evidence indicating that deficits in some aspects of executive functioning may at least partially underlie deficits in social cognition following TBI, and thus help explain the prevalence of social dysfunction in TBI.

Cognitive and psychosocial correlates of alexithymia following traumatic brain injury.

Changes in emotional and social behaviour are considered to be amongst the most common and debilitating consequences of traumatic brain injury (TBI). Little is known of the effects of TBI on alexithymia, which refers to impairment in aspects of understanding emotions. In the current study TBI patients (N=28) were compared with demographically matched healthy controls (N=31) on the Toronto Alexithymia Scale-20 (TAS-20), a measure that taps three distinct characteristics of the alexithymia concept; difficulty in identifying emotions, difficulty in describing emotions and externally oriented thinking. Patients and controls also completed measures of anxiety, depression, quality of life, and measures of fluency to assess executive function. Patients showed greater levels of alexithymia, in terms of difficulty identifying emotions and reduced introspection. Difficulty in identifying emotions was associated with poorer quality of life, even when depression and anxiety were controlled. Difficulty in identifying emotions was also uniquely associated with executive function deficits. Thus, although studies typically focus on aspects of cognitive change following head injury, these results lend support to Becerra et al.'s (Becerra, R., Amos, A., & Jongenelis, S. (2002). Organic alexithymia: a study of acquired emotional blindness. Brain Injury, 16, 633-645.) notion of an 'organic alexithymia', and suggest that more attention should be focused upon assessment of emotional change post-head injury.