Nasal Lipopolysaccharide Challenge and Cytokine Measurement Reflects Innate Mucosal Immune Responsiveness

<div><p>Background</p><p><b>P</b>ractical methods of monitoring innate immune mucosal responsiveness are lacking. Lipopolysaccharide (LPS) is a component of the cell wall of Gram negative bacteria and a potent activator of Toll-like receptor (TLR)-4. To measure LPS responsiveness of the nasal mucosa, we administered LPS as a nasal spray and quantified chemokine and cytokine levels in mucosal lining fluid (MLF).</p><p>Methods</p><p>We performed a 5-way cross-over, single blind, placebo-controlled study in 15 healthy non-atopic subjects (n = 14 <i>per protocol</i>). Doses of ultrapure LPS (1, 10, 30 or 100μg/100μl) or placebo were administered by a single nasal spray to each nostril. Using the recently developed method of nasosorption with synthetic adsorptive matrices (SAM), a series of samples were taken. A panel of seven cytokines/chemokines were measured by multiplex immunoassay in MLF. mRNA for intercellular cell adhesion molecule-1 (ICAM-1) was quantified from nasal epithelial curettage samples taken before and after challenge.</p><p>Results</p><p>Topical nasal LPS was well tolerated, causing no symptoms and no visible changes to the nasal mucosa. LPS induced dose-related increases in MLF levels of IL-1β, IL-6, CXCL8 (IL-8) and CCL3 (MIP-1α) (AUC at 0.5 to 10h, compared to placebo, p<0.05 at 30 and 100μg LPS). At 100μg LPS, IL-10, IFN-α and TNF-α were also increased (p<0.05). Dose-related changes in mucosal ICAM-1 mRNA were also seen after challenge, and neutrophils appeared to peak in MLF at 8h. However, 2 subjects with high baseline cytokine levels showed prominent cytokine and chemokine responses to relatively low LPS doses (10μg and 30μg LPS).</p><p>Conclusions</p><p>Topical nasal LPS causes dose-dependent increases in cytokines, chemokines, mRNA and cells. However, responsiveness can show unpredictable variations, possibly because baseline innate tone is affected by environmental factors. We believe that this new technique will have wide application in the study of the innate immune responses of the respiratory mucosa.</p><p>Key Messages</p><p>Ultrapure LPS was used as innate immune stimulus in a human nasal challenge model, with serial sampling of nasal mucosal lining fluid (MLF) by nasosorption using a synthetic absorptive matrix (SAM), and nasal curettage of mucosal cells. A dose response could be demonstrated in terms of levels of IL-1β, IL-6, CXCL8 and CCL3 in MLF, as well as ICAM-1 mRNA in nasal curettage specimens, and levels of neutrophils in nasal lavage. Depending on higher baseline levels of inflammation, there were occasional magnified innate inflammatory responses to LPS.</p><p>Trial Registration</p><p>Clinical Trials.gov <a href="https://clinicaltrials.gov/ct2/show/NCT02284074?term=nasal+lipopolysaccharide&rank=1" target="_blank">NCT02284074</a></p></div

Somatic Symptom Disorder, Conversion Disorder, and Chronic Pain: Pediatric Clinician Perspectives

Objectives: The appropriateness and relevance of somatic symptom disorder and conversion disorder (American Psychiatric Association, 2013) diagnoses are in question as they relate to pediatric chronic pain. This survey-based study on pediatric chronic pain explores how these psychiatric diagnoses are made and perceived and how treatment of patients is approached by Canadian health care professionals working in pediatric chronic pain clinics. Method: Health care professionals (N = 50) completed the survey, which contained both qualitative and quantitative items. Results: Of participants, 88% reported moderate/advanced training in pain, whereas only 26% reported moderate/advanced training in somatic symptom disorder and conversion disorder. Somatic symptom disorder and conversion disorder were reportedly diagnosed in approximately 17% and 5% of young people with chronic pain, respectively; however, overall, the participants were not confident or only slightly confident when diagnosing these disorders. There were no major differences in the reported interventions used to treat pain, somatic symptom disorder, or conversion disorder. Conclusions: These results highlight the need for standardized training in pain and psychiatric assessment, diagnosis, and treatment; diagnostic guidelines; and how to best provide this training to health care staff who work with young people with chronic pain

Urban Malaria Scheme - Past, Present & Future

Urban Malaria, as a specific problem in India, was first recognized in 1969, after an in- depth review of the situation of malaria in India was done by Madhok Committee. However, main malaria vector in urban areas of India is An. stephensi, which was first recognized in early part of the 20th century. Many of the local bodies carrying out anti- larval operations earlier failed to continue the same due to paucity of funds. During that time, malaria in urban areas was not considered as a major problem because the epidemics recorded earlier in Bombay, Delhi, Lucknow etc. could immediately be contained. Based on recommendations of Madhok Committee to control malaria in urban areas, the Urban Malaria Scheme (UMS) was launched in the country in 1971–7 2. A large scale migration of population, creation of slum clusters, construction activities and water storage practices have contributed in the establishment of malaria foci. Urban malaria control is therefore based on source reduction, larviciding, minor engineering interventions, legislative measures, building bye laws and limited spraying of indoor residual spray in peri- urban and jhuggies. Initially 131 towns with a population of > 40,000 and reporting > 2 API were included in the UMS. Later in 1978, due to merger of National Filaria control programme, 206 towns were also brought under vector control support and the preventive measures against malaria vector breeding in clean water as well as filaria vector breeding in polluted water were integrated. Though there has been a well structured separate programme for prevention and control of malaria in urban areas since 1971, the priority attached to it and its implementation has been an issue primarily because it is the responsibility of the local body. Moreover, in recent past, upsurge in cases of Dengue and Chikungunya in urban areas has diverted attention on both disease surveillance and vector control. Coincidently, the vectors of malaria and dengue/ Chikungunya in urban areas breed in clean water and control strategies are same but surveillance with limited resources has affected in assessing the actual magnitude of problem. This has resulted in upsurge in cases and deaths due to malaria in Mumbai during 2010. Historical background and future vision of Urban Malaria Scheme has been discussed in present article

Evaluation of Direct Rapid Immunohistochemistry Test (DRIT) for Postmortem Diagnosis of Rabies

Direct fluorescent antibody test (DFAT) is considered as the gold standard for diagnosis of rabies in infected mammals as it has high sensitivity and specificity. However, high cost and technical demand limits its utilization, particularly in developing countries including India. Therefore, in this study we evaluated recently developed direct rapid immunohistochemistry test (DRIT) for diagnosis of rabies in comparison with the DFAT. A total of 109 brain samples received during the period of 6 years from different regions of India were tested following standard protocol. The results showed 100% correlation between the two tests.

Development of SSR markers and construction of a linkage map in jute

Jute is an important natural fibre crop, which is only second to cotton in its importance at the global level. It is mostly grown in Indian subcontinent and has been recently used for the development of genomics resources. We recently initiated a programme to develop simple sequence repeat markers and reported a set of 2469 SSR that were developed using four SSR-enriched libraries (Mir et al. 2009). In this communication, we report an additional set of 607 novel SSR in 393 SSR containing sequences. However, primers could be designed for only 417 potentially useful SSR. Polymorphism survey was carried out for 374 primer pairs using two parental genotypes (JRO 524 and PPO4) of a mapping population developed for fibre fineness; only 66 SSR were polymorphic. Owing to a low level of polymorphism between the parental genotypes and a high degree of segregation distortion in recombinant inbred lines, genotypic data of only 53 polymorphic SSR on the mapping population consisting of 120 RIL could be used for the construction of a linkage map; 36 SSR loci were mapped on six linkage groups that covered a total genetic distance of 784.3 cM. Hopefully, this map will be enriched with more SSR loci in future and will prove useful for identification of quantitative trait loci/genes for molecular breeding involving improvement of fibre fineness and other related traits in jute

A tablet computer-assisted motor and language skills training programme to promote communication development in children with autism: development and pilot study

Autism is a heterogenous condition, encompassing many different subtypes and presentations. Of those people with autism who lack communicative speech, some are more skilled at receptive language than their expressive difficulty might suggest. This disparity between what can be spoken and what can be understood correlates with motor and especially oral motor abilities, and thus may be a consequence of limits to oral motor skill. Point OutWords, tablet-based software targeted for this subgroup, builds on autistic perceptual and cognitive strengths to develop manual motor and oral motor skills prerequisite to communication by pointing or speaking. Although typical implementations of user-centred design rely on communicative speech, Point OutWords users were involved as co-creators both directly via their own nonverbal behavioural choices and indirectly via their communication therapists’ reports; resulting features include vectorised, high-contrast graphics, exogenous cues to help capture and maintain attention, customisable reinforcement prompts, and accommodation of open-loop visuomotor control