Integrating Blood Collection Within Household Surveys: Lessons Learned From Nesting a Measles and Rubella Serological Survey Within a Post-Campaign Coverage Evaluation Survey in Southern Province, Zambia

Age-specific population immunity to many vaccine-preventable diseases can be measured using serological surveys. However, stand-alone serological surveys are infrequently conducted in low- and middle-income countries because of costs, operational challenges, and potential high refusal rates for blood collection. Nesting a serosurvey within a household cluster survey may overcome some of these challenges. We share lessons learned from nesting a serosurvey within a measles and rubella vaccination post-campaign coverage evaluation survey (PCES). In 15 of the 26 PCES clusters in Southern Province, Zambia, we collected dried blood spots from 581 participants aged 9 months and older. Household participation rates for the main PCES were higher in the serosurvey clusters (86%) than PCES-only clusters (71%), suggesting that a serosurvey can be successfully integrated without adversely affecting PCES participation. Among households that participated in the PCES, 80% also participated in the serosurvey and 86% of individuals available in the household provided a blood sample for the serosurvey. Substantial planning and coordination, additional staff training, and community mobilization were critical to the success of the serosurvey. Most challenges stemmed from using different data collecting tools and teams for the serosurvey and PCES. A more efficient design would be to fully integrate the serosurvey by adding blood collection and additional questions to the PCES

Inhibitory Potential of Prodomain of Plasmodium falciparum Protease Serine Repeat Antigen 5 for Asexual Blood Stages of Parasite

Plasmodium falciparum serine repeat antigen 5 (SERA5) is a target for both drug and vaccine intervention against malaria. SERA5 is secreted in the parasitophorous vacuole where it is proteolytically processed before schizont rupture. Among the processed products is a 50.8-kDa central domain of the protease, which possesses chymotrypsin-like activity and consists of a 28.9-kDa catalytic domain with a 21.9-kDa N-terminal prodomain, which remain attached together. Because SERA5 has been implicated in merozoite egress from host erythrocytes, the effect of the prodomain and a heptapeptide derived from its C-terminus spanning from D560 to F566 (DNSDNMF) on parasite growth was studied. When E. coli-expressed prodomain was incubated with parasite culture, a significant delay in transition from schizont to ring stages was observed up to nanomolar concentrations. The peptide, DNSDNMF also showed similar effects but at nearly 1000-fold higher concentrations. The peptide was also found to interact with the catalytic domain. These data demonstrate the crucial role of SERA5 prodomain for the egress process. Given the inhibitory potential of the prodomain for the parasite, we suggest that peptidomimetic inhibitors based on SERA5 prodomain sequences can be developed as future therapeutics against malaria

NF-kappaB p65-Dependent Transactivation of miRNA Genes following Cryptosporidium parvum Infection Stimulates Epithelial Cell Immune Responses

Cryptosporidium parvum is a protozoan parasite that infects the gastrointestinal epithelium and causes diarrheal disease worldwide. Innate epithelial immune responses are key mediators of the host's defense to C. parvum. MicroRNAs (miRNAs) regulate gene expression at the posttranscriptional level and are involved in regulation of both innate and adaptive immune responses. Using an in vitro model of human cryptosporidiosis, we analyzed C. parvum-induced miRNA expression in biliary epithelial cells (i.e., cholangiocytes). Our results demonstrated differential alterations in the mature miRNA expression profile in cholangiocytes following C. parvum infection or lipopolysaccharide stimulation. Database analysis of C. parvum-upregulated miRNAs revealed potential NF-κB binding sites in the promoter elements of a subset of miRNA genes. We demonstrated that mir-125b-1, mir-21, mir-30b, and mir-23b-27b-24-1 cluster genes were transactivated through promoter binding of the NF-κB p65 subunit following C. parvum infection. In contrast, C. parvum transactivated mir-30c and mir-16 genes in cholangiocytes in a p65-independent manner. Importantly, functional inhibition of selected p65-dependent miRNAs in cholangiocytes increased C. parvum burden. Thus, we have identified a panel of miRNAs regulated through promoter binding of the NF-κB p65 subunit in human cholangiocytes in response to C. parvum infection, a process that may be relevant to the regulation of epithelial anti-microbial defense in general

Comparative genome analysis of two Cryptosporidium parvum isolates with different host range

Parasites of the genus Cryptosporidium infect the intestinal and gastric epithelium of different vertebrate species. Some of the many Cryptosporidium species described to date differ with respect to host range; whereas some species’ host range appears to be narrow, others have been isolated from taxonomically unrelated vertebrates. To begin to investigate the genetic basis of Cryptosporidium host specificity, the genome of a C. parvum isolate belonging to a sub-specific group found exclusively in humans was sequenced and compared to the reference C. parvum genome representative of the zoonotic group. Over 12,000 single-nucleotide polymorphisms (SNPs), or 1.4 SNP per kilobase, were identified. The genome distribution of SNPs was highly heterogeneous, but non-synonymous and silent SNPs were similarly distributed. On many chromosomes, the most highly divergent regions were located near the ends. Genes in the most diverged regions were almost twice as large as the genome-wide average. Transporters, and ABC transporters in particular, were over-represented among these genes, as were proteins with predicted signal peptide. Possibly reflecting the presence of regulatory sequences, the distribution of intergenic SNPs differed according to the function of the downstream open reading frame. A 3-way comparison of the newly sequenced anthroponotic C. parvum, the reference zoonotic C. parvum and the human parasite C. hominis identified genetic loci where the anthroponotic C. parvum sequence is more similar to C. hominis than to the zoonotic C. parvum reference. Because C. hominis and anthroponotic C. parvum share a similar host range, this unexpected observation suggests that proteins encoded by these genes may influence the host range

Mucins and Pathogenic Mucin-Like Molecules Are Immunomodulators During Infection and Targets for Diagnostics and Vaccines

Mucins and mucin-like molecules are highly O-glycosylated proteins present on the cell surface of mammals and other organisms. These glycoproteins are highly diverse in the apoprotein and glycan cores and play a central role in many biological processes and diseases. Mucins are the most abundant macromolecules in mucus and are responsible for its biochemical and biophysical properties. Mucin-like molecules cover various protozoan parasites, fungi and viruses. In humans, modifications in mucin glycosylation are associated with tumors in epithelial tissue. These modifications allow the distinction between normal and abnormal cell conditions and represent important targets for vaccine development against some cancers. Mucins and mucin-like molecules derived from pathogens are potential diagnostic markers and targets for therapeutic agents. In this review, we summarize the distribution, structure, role as immunomodulators, and the correlation of human mucins with diseases and perform a comparative analysis of mucins with mucin-like molecules present in human pathogens. Furthermore, we review the methods to produce pathogenic and human mucins using chemical synthesis and expression systems. Finally, we present applications of mucin-like molecules in diagnosis and prevention of relevant human diseases

Structural and aesthetic design applications of flexible, thin-film solar cells to power off-grid tensile structures

Thesis: S.M. in Engineering and Management, Massachusetts Institute of Technology, System Design and Management Program, May, 2020Thesis: S.M., Massachusetts Institute of Technology, Department of Mechanical Engineering, May, 2020Cataloged from the official pdf version of thesis. "May 2020."Includes bibliographical references (pages [75]-[77]).Despite global trends in decreasing costs of silicon-based solar panels, the adoption of solar energy solutions as an alternative to fossil fuels has been impeded by high installation and manufacturing costs, as well as challenges in the customization of solar panels for different products and environments. Moreover, silicon-based photovoltaic cells, due to their rigid nature, change the aesthetic of the surfaces on which they are placed and often only provide the singular function of harvesting energy. The current solar energy products function independently from the architecture on which they are installed, making them difficult to blend in with the design and functional requirements of the products and buildings on which they are installed. Fundamentally, the installation costs associated with silicon crystalline PV cells account for a significant percentage of solar energy solutions. This thesis aims to push the boundaries of solar panels to provide the dual functionality of energy harvesting and architectural structure, while either maintaining or improving the aesthetics of the architecture on which they are placed. To achieve this, this research explores a new use case for flexible thin-cell solar panels that includes the use of organic photovoltaic (OPV) and perovskite solar cell technology. Through a product-design approach, this thesis explores use cases where the technology's uniquely-flexible, ultra-thin, lightweight, and low-cost key features are best applied as a solar energy source. Particularly, this research focuses on off-grid architecture with non-rigid roofing structures where fossil fuels are currently used as the primary energy source. Through design research and stakeholder interviews, a key insight that was uncovered was the opportunity to integrate flexible OPV solar cells in glamping and luxury safari camp as an alternative to the current option of diesel fuel. This achieves the goal of providing a clean energy source while maintaining the aesthetic of the luxury camp and the outdoor safari experience.by Juliet Wanjiru Wanyiri.S.M. in Engineering and ManagementS.M.S.M.inEngineeringandManagement Massachusetts Institute of Technology, System Design and Management ProgramS.M. Massachusetts Institute of Technology, Department of Mechanical Engineerin

Leveraging microgravity to investigate earth- And space-based centrifugal casting of wax

A multi-year research effort aimed at increasing understanding of the centrifugal casting process of wax fuels for hybrid chemical propulsion in multiple thermal and gravitational environments is described. As both radiative and convective heat transfer drive the casting process, the suborbital and orbital microgravity environments are critical to disentangling these contributions to heat transfer away from the fuel. The experimental effort comprises testing on multiple platforms, including the ambient atmosphere of the laboratory, as well as various mobile microgravity platforms. Testing onboard reduced-gravity aircraft facilitates increased understanding of how these types of fluids perform in the microgravity environment, while a suborbital spaceflight and orbital platform under standard atmosphere allow for longer-term observation of natural convection sans buoyancy. An orbital platform subjected to the space environment facilitates understanding of the contribution of radiation to the heat transfer away from the liquid fuel. Each progressive testing environment requires updates to the experimental setup in order to accommodate respective physical and electrical constraints which are described in detail herein. An image analysis routine was developed in order to automate post-processing and determine the solidification front speed for each test. A rotation rate actuation routine is in development which aims to improve the accuracy of the centrifuge control system by leveraging electromagnetic sensing and feeding back rotation rate measurements to the motor driver. Preliminary modeling work was conducted which aims to elucidate the fundamental physics of the centrifugal casting problem; specifically, the impact of rotation rate, material properties, and environmental conditions on the heat transfer and fluid mechanics which constitute the larger casting problem. Both paraffin wax - a solid fuel with two decades of heritage - and the more novel beeswax are considered in this study

Role of CpSUB1, a subtilisin-like protease, in Cryptosporidium parvum infection in vitro.

The apicomplexan parasite Cryptosporidium is a significant cause of diarrheal disease worldwide. Previously, we reported that a Cryptosporidium parvum subtilisin-like serine protease activity with furin-type specificity cleaves gp40/15, a glycoprotein that is proteolytically processed into gp40 and gp15, which are implicated in mediating infection of host cells. Neither the enzyme(s) responsible for the protease activity in C. parvum lysates nor those that process gp40/15 are known. There are no furin or other proprotein convertase genes in the C. parvum genome. However, a gene encoding CpSUB1, a subtilisin-like serine protease, is present. In this study, we cloned the CpSUB1 genomic sequence and expressed and purified the recombinant prodomain. Reverse transcriptase PCR analysis of RNA from C. parvum-infected HCT-8 cells revealed that CpSUB1 is expressed throughout infection in vitro. In immunoblots, antiserum to the recombinant CpSUB1 prodomain revealed two major bands, of approximately 64 kDa and approximately 48 kDa, for C. parvum lysates and proteins "shed" during excystation. In immunofluorescence assays, the antiserum reacted with the apical region of sporozoites and merozoites. The recombinant prodomain inhibited protease activity and processing of recombinant gp40/15 by C. parvum lysates but not by furin. Since prodomains are often selective inhibitors of their cognate enzymes, these results suggest that CpSUB1 may be a likely candidate for the protease activity in C. parvum and for processing of gp40/15. Importantly, the recombinant prodomain inhibited C. parvum infection of HCT-8 cells. These studies indicate that CpSUB1 plays a significant role in infection of host cells by the parasite and suggest that this enzyme may serve as a target for intervention