Intranasally administered extracellular vesicles from human induced pluripotent stem cell-derived neural stem cells quickly incorporate into neurons and microglia in 5xFAD mice.

Introduction: Extracellular vesicles (EVs) released by human-induced pluripotent stem cell (hiPSC)-derived neural stem cells (NSCs) have robust antiinflammatory and neurogenic properties due to therapeutic miRNAs and proteins in their cargo. Hence, hiPSC-NSC-EVs are potentially an excellent biologic for treating neurodegenerative disorders, including Alzheimer's disease (AD). Methods: This study investigated whether intranasally (IN) administered hiPSC-NSC-EVs would quickly target various neural cell types in the forebrain, midbrain, and hindbrain regions of 3-month-old 5xFAD mice, a model of ß-amyloidosis and familial AD. We administered a single dose of 25 × 109 hiPSC-NSC-EVs labeled with PKH26, and different cohorts of naïve and 5xFAD mice receiving EVs were euthanized at 45 min or 6 h post-administration. Results: At 45 min post-administration, EVs were found in virtually all subregions of the forebrain, midbrain, and hindbrain of naïve and 5xFAD mice, with predominant targeting and internalization into neurons, interneurons, and microglia, including plaque-associated microglia in 5xFAD mice. EVs also came in contact with the plasma membranes of astrocytic processes and the soma of oligodendrocytes in white matter regions. Evaluation of CD63/CD81 expression with the neuronal marker confirmed that PKH26 + particles found within neurons were IN administered hiPSC-NSC-EVs. At 6 h post-administration, EVs persisted in all cell types in both groups, with the distribution mostly matching what was observed at 45 min post-administration. Area fraction (AF) analysis revealed that, in both naïve and 5xFAD mice, higher fractions of EVs incorporate into forebrain regions at both time points. However, at 45 min post-IN administration, AFs of EVs within cell layers in forebrain regions and within microglia in midbrain and hindbrain regions were lower in 5xFAD mice than naïve mice, implying that amyloidosis reduces EV penetrance. Discussion: Collectively, the results provide novel evidence that IN administration of therapeutic hiPSC-NSC-EVs is an efficient avenue for directing such EVs into neurons and glia in all brain regions in the early stage of amyloidosis. As pathological changes in AD are observed in multiple brain areas, the ability to deliver therapeutic EVs into various neural cells in virtually every brain region in the early stage of amyloidosis is attractive for promoting neuroprotective and antiinflammatory effects.

Advanced virtual mentoring of dentists in oral cancer screening and tobacco cessation - An interventional study.

Context: Oral cancer is the third common cancer in India. Its mortality can be reduced through early detection and tobacco cessation ideally by dentists owing to their forte of work. Aim: This study was conducted to discuss effectiveness of an advanced tele-mentoring programme in oral cancer screening and tobacco cessation for dentists across India. Settings and Design: Online, interventional study. Methods and Material: The 14-week long training programme with 52 participants/spokes from across India had weekly hour-long online sessions comprising of an expert-led didactic and case discussions by spokes. Online evaluation (pre- and post-training, post-session), weekly and post-one-year feedback were conducted. Successful spokes attended a hands-on workshop subsequently. Statistical Analysis Used: One and independent sample t-tests determined the significance of the evaluation scores of the participants. Findings on attitudes and practice-related questions are presented as simple percentages. Results: A notable increase in the overall and per-session mean knowledge score, and confidence in oral cancer screening was observed. Many participants started these services at their clinics, thereby reducing further referrals, and were also motivated to spread community awareness about the same. Conclusion: This tele-mentoring programme, based on the novel Extension for Community Healthcare Outcomes model, is the first oral cancer screening training programme for dentists. This model-comprising of expert didacts, case discussions, and significant spoke-expert interaction-is a promising best-practices tool for reducing the disparity in knowledge and skills regarding oral cancer prevention among dentists across different locations. This would enable these most appropriate healthcare providers to contribute toward the overall goal of oral cancer prevention.

Barriers and facilitators in implementing population based common cancer screening through community health workers.

Population based cancer screening was initiated in India in 2016 owing to an increased burden of cancers. A feasibility health system study was done by utilising community health workers (CHWs) to conduct the cancer screening. The current study is a qualitative study to elicit the barriers and facilitators in implementing population based cancer screening through CHWs. The study was conducted at three subcentres of Dholai block of Cachar district, Assam, India and Cachar Cancer Hospital and Research Center, Silchar. The participants of the study were CHWs, master trainer nurses and women from community. Three focus group discussions (FGDs) and one in-depth interview (IDI) were conducted at the provider level and seven IDIs of women from the community. The FGDs and IDIs were audio recorded after taking verbal consent from the participants. The verbatims were prepared following translation and transcription and data analysis using ATLAS ti ver 8. The major barrier faced by the community was a lack of motivation to get screened which stemmed from various factors such as personal beliefs, attitudes and fear. The major facilitators were accessibility of tests, family support and CHWs as screening service providers. The major barriers for CHWs were difficulty in motivating the community, lack of support from supervisors and lack of motivation to work. The major facilitators were convenience of screening during home visits, empowerment, skill enhancement and teamwork. Population based cancer screening was a new concept for the community under study. Cancer screening by CHWs was well accepted by the community. Awareness generation among the community was a major factor in improving screening coverage. The study highlights that training and motivation of CHWs can improve the uptake of cancer screening services. CHWs felt empowered with the new skills imparted and were able to carry out screening.

Capacity Building of Gynecologists in Cancer Screening Through Hybrid Training Approach.

Training health care professionals (HCPs) is one of the most challenging and key factors for the success of a cancer screening program. In order to make this onerous task possible, a hybrid training model, combining the online knowledge-sharing tool of ECHO (Extension of Community Health Outcomes) and in-person training, was proposed by the National Institute of Cancer Prevention and Research (NICPR). The main aim of this article is disseminating our experience on the effectiveness of this hybrid model in training health care providers in cancer prevention. A group of gynecologists was trained using a structured curriculum in cervical and breast cancer screening through a 14-week online course, followed by a three-day in-person training (group A). To analyze the effectiveness of this model, a group of gynecologists who were not part of the online course were enrolled for face-to-face training (group B). All the participants were offered pre- and post-training questionnaires and a pictorial quiz. Group A participants had 60% and 40% more knowledge in cervical and breast cancer screening, respectively, compared with group B before the in-person training. Though group B demonstrated a 51% increase in knowledge post-training, group A performed 26% better than group B in the pictorial quiz-demonstrating better knowledge acquisition. This hybrid training model, when employed in capacity building in cancer screening among gynecologists, works very effectively in improving knowledge and skill set in cancer screening. This can be a potent tool for the government for efficient training of HCPs in cancer screening.

Project ECHO: a Potential Best-Practice Tool for Training Healthcare Providers in Oral Cancer Screening and Tobacco Cessation.

The oral cancer pandemic and inadvertent tobacco consumption have rendered timely oral cavity screening and tobacco cessation essential, skills which most healthcare providers (HCPs) lack. Project "Extension for Community Healthcare Outcomes" (ECHO) is a proven best-practice tool for virtual telementoring of primary care providers by experts at academic health centers, in managing complex medical conditions in rural, expert-deficient setups. For the first time, our organization in India has utilized this method for training HCPs in oral cancer screening, across the country and abroad. The program comprised eight, weekly, hour-long sessions, on oral cancer screening and tobacco cessation, hosted online by our Organization (hub) through the Zoom web-conferencing application, with 48 HCPs (spokes) attending from their respective locations (pan-India, n = 47; Libya, n = 1). Each session comprised one expert-led didactic and two participant-led case presentations, culminating with educative discussions. Participants filled out online, program-evaluation (pre and post) questionnaires having 10 similar, multiple-choice questions each (score for every correct response = 1); total responses were later statistically analyzed. Lesser participants completed the post-evaluation questionnaire which could be due to it being optional, their busy schedule, or apprehension of being assessed. The program evaluation results illustrate a significant knowledge gain among participants regarding oral cancer screening and tobacco cessation, i.e., from a mean knowledge score of 6.7 in pre-evaluation to 7.4 in post-evaluation (p < 0.05). Thus, the ECHO model can be utilized as a convenient, cost-effective, large-scale, best-practice, telementoring tool for training HCPs in oral cancer screening and tobacco cessation, especially in populous, resource-deficient countries.

Altered subcellular localization of the NeuN/Rbfox3 RNA splicing factor in HIV-associated neurocognitive disorders (HAND).

The anti-NeuN antibody has been widely used for over 15 years to unambiguously identify post-mitotic neurons in the central nervous system of a wide variety of vertebrates including mice, rats and humans. In contrast to its widely reported nuclear localization, we found significantly higher NeuN reactivity in the cytoplasm of neurons in brain sections from HIV-infected individuals with cognitive impairment compared to controls. The protein target of anti-NeuN antisera was recently identified as the neuron-specific RNA splicing factor, Rbfox3, but its significance in diseases affecting the brain has not been previously reported. RNA splicing occurs in the nucleus hence, the altered localization of RbFox3 to the cytoplasm may lead to the downregulation of neuronal gene expression.

A consensus surface activation marker signature is partially dependent on human immunodeficiency virus type 1 Nef expression within productively infected macrophages.

BACKGROUND: The high prevalence of HIV-associated comorbidities including neurocognitive disorder, high levels of residual inflammatory mediators in the plasma and cerebrospinal fluid and the resurgence of HIV replication upon interruption of antiviral treatment in HIV-1 infected individuals, strongly suggests that despite therapy HIV persists in its cellular targets which include T-lymphocytes and cells of the myeloid lineage. These reservoirs present a major barrier against eradication efforts. Knowledge of the molecular mechanisms used by HIV to modulate innate macrophage immune responses and impair viral clearance is quite limited. To explore the role of HIV in potentially modulating macrophage function through changes in protein expression, we used single-cell analyses with flow cytometry to determine whether, in unpolarized cultures, macrophage surface marker phenotype was altered by HIV infection in a manner that was independent of host genetic background. RESULTS: These analyses revealed that at several time points post-infection, GFP + HIV-infected macrophages were significantly enriched in the CD14+ fraction (3 to 5-fold, p = .0001) compared to bystander, or uninfected cells in the same culture. However, the enrichment and higher levels of CD14 on HIV expressing macrophages did not depend on the production of HIV Nef. Sixty to eighty percent of macrophages productively infected with HIV after day 28 post-infection were also enriched in the population of cells expressing the activation markers CD69 (2 to 4-fold, p < .0001) and CD86 (2 to 4-fold, p < .0001 ) but suppressed amounts of CD68 (3 to 10-fold, p < .0001) compared to bystander cells. Interestingly, there was no enrichment of CD69 on the surface of HIV producing cells that lacked Nef or expressed a variant of Nef mutated in its SH3-binding domain. CONCLUSIONS: These findings suggest that HIV actively regulates the expression of a subset of surface molecules involved in innate and inflammatory immune signaling in primary human macrophages through Nef-dependent and Nef-independent mechanisms acting within productively infected cells.