Recently activated CD4 T cells in tuberculosis express OX40 as a target for host-directed immunotherapy.

After Mycobacterium tuberculosis (Mtb) infection, many effector T cells traffic to the lungs, but few become activated. Here we use an antigen receptor reporter mouse (Nur77-GFP) to identify recently activated CD4 T cells in the lungs. These Nur77-GFPHI cells contain expanded TCR clonotypes, have elevated expression of co-stimulatory genes such as Tnfrsf4/OX40, and are functionally more protective than Nur77-GFPLO cells. By contrast, Nur77-GFPLO cells express markers of terminal exhaustion and cytotoxicity, and the trafficking receptor S1pr5, associated with vascular localization. A short course of immunotherapy targeting OX40+ cells transiently expands CD4 T cell numbers and shifts their phenotype towards parenchymal protective cells. Moreover, OX40 agonist immunotherapy decreases the lung bacterial burden and extends host survival, offering an additive benefit to antibiotics. CD4 T cells from the cerebrospinal fluid of humans with HIV-associated tuberculous meningitis commonly express surface OX40 protein, while CD8 T cells do not. Our data thus propose OX40 as a marker of recently activated CD4 T cells at the infection site and a potential target for immunotherapy in tuberculosis.

18†Engaging with young people to improve research, services and workforce development: eye-YPAG and 'visually' workshops.

Involving children and young people (CYP) in service and research design improves quality and accessibility. Running events in schools to invite CYP to volunteer and explore careers in the NHS may contribute to uptake of training posts and developing the NHS workforce.Here we evaluate two activities with CYP, our Young Person's Advisory Group for research (eye-YPAG) and our workshop for secondary schools, 'visually'.We evaluated eye-YPAG in focus groups and online surveys with group members, parents/carers, researchers, facilitators and funders. We conducted thematic analysis and descriptive statistics. To evaluate 'visually', we monitored the numbers of workshops and young people applying for volunteering roles. We asked those who started working with us about their experience.eye-YPAG members valued social and creative aspects as well as learning about research and developing skills and confidence. Researchers reported that CYP gave novel suggestions, modifying research plans, and that their different perspective was helpful in making research more relevant for children and families.Over 6 months, we held 15 'visually' workshops in secondary schools. Ninety students applied for volunteering roles, and 20 have completed the Human Resources onboarding process. Young volunteers report that this work has increased their confidence and that they have gained insights into how a hospital works. One is considering training to become an orthoptist.Both eye-YPAG and 'visually' are available to all eye researchers and units in the UK and can facilitate outreach activities.

Engaging male students with mental health support: a qualitative focus group study.

BACKGROUND: Males are less likely to seek help for mental health difficulties compared to females. Despite considerable interest, a paucity of evidence-based solutions exists to address this. Concerns about students' mental health has led to the United Kingdom's Department of Education to make this a priority. Studies have shown that male students hold more negative attitudes towards the use of psychological services compared to female students and are less likely to seek help. A major concern is that male students make up 69% of university suicides, which is often associated with lower rates of help-seeking. This focus group study therefore sought to identify potential approaches that would be relevant to improving mental health help-seeking in male students. METHODS: Three focus groups comprising of 24 male students at a London University were conducted. Participants were asked questions exploring: the barriers to seeking help, what would encourage help-seeking, how an appropriate intervention should be designed, and how to publicise this intervention to male students. Thematic analysis was conducted to evaluate participants responses. RESULTS: Five distinct themes were identified. These were: 1) protecting male vulnerability, 2) providing a masculine narrative of help-seeking, 3) differences over intervention format, 4) difficulty knowing when and how to seek help, and 5) strategies to sensitively engage male students. CONCLUSIONS: These themes represent important considerations that can be used, together with the existing literature about male help-seeking, to develop more male friendly interventions that are suitable for male students. This could help improve help-seeking attitudes and the uptake of mental health interventions for male students experiencing emotional distress.

Quality of life and functional vision in children treated for cataract-a cross-sectional study.

PurposeChildren with cataract and their families face intensive medical and surgical management, with numerous hospital attendances, topical medications, and surgical procedures, as well as uncertainty about the child's future visual ability, education, and independence. Little is known about the impact on functional visual ability, vision-, and health-related quality of life (VR-, HR-QoL).Patients and methodsSeventy two children aged 2-16 years (mean 8.45, SD 4.1) treated for developmental or secondary cataract and their parents/carers completed three validated instruments measuring functional visual ability, VR-, and HR-QoL: the Cardiff Visual Ability Questionnaire for Children (CVAQC), Impact of Vision Impairment for Children (IVI-C), and PedsQL V 4.0.ResultsAll scores are markedly reduced: median (interquartile range (IQR)) CVAQC score -1.42 (-2.28 to -0.03), mean (SD) IVI-C score 65.67 (16.91), median (IQR) PedsQL family impact score 75 (56.94-88.19), parent report 71.74 (51.98-88.5), self-report 76.09 (61.96-89.13). Psychosocial PedsQL subscores are lower than physical subscores. Parent-completed tools (PedsQL family and parent report) state greater impact on HR-QoL than tools completed by children/young people, particularly in teenagers. Older children/young people have higher functional visual ability scores than younger children.ConclusionsCataract has a marked a long-term impact on functional visual ability and quality of life of children and young people, with HR-QoL affected to degrees reported in children with severe congenital cardiac defects or liver transplants.

Validation of a computerised logMAR visual acuity measurement system (COMPlog): comparison with ETDRS and the electronic ETDRS testing algorithm in adults and amblyopic children.

BACKGROUND/AIM: The COMPlog clinical visual acuity measuring system is being developed for both routine and research use. This study aimed to validate its performance in amblyopic children and both normal and diseased adults against the gold standard ETDRS chart and the E-ETDRS computerised acuity measurement algorithm. METHOD: Timed test and retest fully interpolated five letters per line logMAR visual acuity measurements were taken for 70 adults and 59 amblyopic children using the ETDRS chart and the COMPlog visual acuity measurement system. 39 of the adults also underwent computerised acuity testing using the E-ETDRS testing algorithm. The tested adults included normals as well as subjects with a range of ocular diseases. The methods of Bland and Altman were employed with test-retest variability (TRV) expressed as 95% confidence limits for agreement. RESULTS: No significant bias was observed between the gold standard ETDRS acuity measurements and those taken with either COMPlog or E-ETDRS. TRVs of +/-0.12 logMAR and +/-0.10 logMAR were respectively found for COMPlog measurements in the amblyopic children and adult groups compared with +/-0.12 logMAR for the ETDRS chart in both groups. The TRV of the E-ETDRS system was slightly greater at +/-0.16 logMAR. Median testing times for COMPlog and ETDRS were 95 and 85 s and 66 and 56 s respectively in the paediatric and adult groups and 120 s for the E-ETDRS measurements on adults. DISCUSSION: COMPlog measurements agree well with and are similarly reliable to the gold standard ETDRS chart with comparable test times. E-ETDRS algorithm measurements took approximately twice as long.

Sulphated compounds attenuate beta-amyloid toxicity by inhibiting its association with cells.

Agents that interfere with the toxic effects of beta-amyloid protein may be therapeutically useful against Alzheimer's disease. We reported recently that several sulphated glycosaminoglycans and sulphonated dyes attenuate the toxic effects of beta-amyloid fragments beta 25-35 and beta 1-40 in two clonal cell lines. We now demonstrate that this protective effect is due to interference with beta-amyloid cell association rather than effects on beta-amyloid structure. Using an enzyme-linked immunoabsorbance assay to detect cell-associated beta 1-40, we found in a range of compounds a strong correlation between inhibition of HeLa cell association of beta 1-40 and attenuation of cellular toxicity as measured by inhibition of 3-[4,5-dimethylthia-zol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) reduction. In contrast, effects on peptide structure, as measured by Congo red binding, were generally inconsistent with the attenuating effects of the compounds on cellular toxicity. These results suggest that by binding beta-amyloid these agents prevent its interaction with cells.

Sulfonated dyes attenuate the toxic effects of beta-amyloid in a structure-specific fashion.

We recently reported that several sulfate-containing glycosaminoglycans, a class of compounds associated with the beta-amyloid plaques of Alzheimer's disease, attenuate the toxic effects of beta-amyloid fragments beta 25-35 and beta 1-40. The amyloid-binding sulfonated dye Congo Red was shown to have a similar effect. Using two clonal cell lines, we now demonstrate that several sulfonated dyes attenuate beta-amyloid toxicity and that the protective effect appears specific for compounds whose sulfonate groups can interact with the beta-pleated structure of aggregated amyloid. These results suggest that by binding beta-amyloid these compounds may prevent toxic interactions of the peptide with cells.

The intracellular component of cellular 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) reduction is specifically inhibited by beta-amyloid peptides.

In vitro cell culture model systems for investigating the biochemical mechanisms involved in the neurodegenerative actions of beta-amyloid peptide (beta-AP) have been established. Using rat pheochromocytoma PC12 or human epitheloid HeLa cell lines, submicromolar concentrations of the beta-AP fragments beta 1-40, beta 1-39, and beta 25-35, but not beta 1-28, were found to inhibit the reduction of the redox dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). In both cell lines, the beta-AP-sensitive component represented approximately 70% of total cellular MTT reduction. When the reduction of a series of structurally related dyes was compared with that of MTT, the reduction of 3 alpha-naphthyl-2-phenyl-5-(4-nitrophenyl)-2H-tetrazolium chloride (NTV) was also found to be sensitive to beta 25-35, but that of seven other redox dyes was not. A property common to MTT and NTV is that they are both readily taken up into PC12 and HeLa cells and do not require an artificial electron coupling agent to be reduced. Microscopic analysis of MTT-formazan product formation in PC12 and HeLa cells following beta 25-35 treatment revealed that it was the intracellular component of the reduction of this dye that was abolished. These results support the hypothesis that the cellular reduction of MTT represents a specific indicator of the initial events underlying the mechanism of beta-AP toxicity.

Sulfated glycosaminoglycans and dyes attenuate the neurotoxic effects of beta-amyloid in rat PC12 cells.

Glycosaminoglycan (GAG)-containing proteoglycans are associated with the neuritic plaques and cerebrovascular beta-amyloid deposits of Alzheimer's disease as well as with the amyloid deposits of prion and other disorders. GAGs and other sulfate-containing compounds have previously been shown to bind beta-amyloid peptide in vitro, suggesting possible effects of beta-amyloid deposition and/or toxicity in vivo. Using reduction of the redox dye 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) to measure beta-amyloid neurotoxicity in rat pheochromocytoma PC12 cells, several polysulfated GAGs and synthetic sulfate-containing compounds were found to attenuate the neurotoxic effects of beta-amyloid fragments beta 25-35 and beta 1-40. These results suggest that by binding beta-amyloid these compounds may prevent toxic interactions of the peptide with cells.

A study of aqueous and serum levels of ceftazidime following subconjunctival administration.

Patients undergoing cataract surgery may develop an infective endophthalmitis postoperatively which may result in the loss of an eye. This study was carried out to measure aqueous humour levels and to assess patients' tolerance of ceftazidime, a potent antipseudomonal cephalosporin, given subconjunctivally. Eighteen patients received 125 mg ceftazidime subconjunctivally before they underwent routine cataract surgery. A further two patients received 62.5 mg subconjunctivally. The results show good penetration into the aqueous humour well above the minimum inhibitory concentrations (MICs) of possible pathogens. There were no postoperative infections, no local irritation, and no systemic side effects.