Peridomestic conditions affect La Crosse virus entomological risk by modifying the habitat use patterns of its mosquito vectors.

Anthropogenic land-use change may affect the transmission risk for endemic vector-borne diseases such as La Crosse encephalitis. In this study, we applied a comparative ecological approach to evaluate differences in vector species abundance, gonotrophic status, and environmental variables among six residential habitats (historical case houses) and six paired adjacent forest patches in a La Crosse virus endemic area of North Carolina. A total of 93,158 container Aedes spp. eggs were obtained by ovistrips and 1,040 resting mosquito adults were collected by large-bore aspiration from 10 June through 8 October, 2010. At sites characterized by high densities of artificial containers, the totals of eggs and adult mosquitoes were higher in the peridomestic plots. At sites characterized by lower densities of artificial containers, the totals of eggs and adult mosquitoes were higher in the forested plots. Although Aedes triseriatus, the primary La Crosse virus vector, was more commonly found in forested habitat overall, in sites characterized by high density of artificial containers, it was found in higher abundance in the peridomestic habitat. Similarly, the proportion of gonotrophically active (parous and gravid) mosquitoes was higher in the peridomestic habitat for sites with highartificial container density. Our study suggests that La Crosse virus transmission risk may be higher at peridomestic habitats with higher densities of artificial containers and thus reinforces the importance of public health measures to improve source reduction efforts.

Control of Persistent Salmonella Infection Relies on Constant Thymic Output Despite Increased Peripheral Antigen-Specific T Cell Immunity.

Recent thymic emigrants are the youngest subset of peripheral T cells and their involvement in combating persistent bacterial infections has not been explored. Here, we hypothesized that CD4+ recent thymic emigrants are essential immune mediators during persistent Salmonella infection. To test this, we thymectomized adult mice either prior to, or during, persistent Salmonella infection. We found that thymic output is crucial in the formation of protective immune responses during the early formation of a Salmonella infection but is dispensable once persistent Salmonella infection is established. Further, we show that thymectomized mice demonstrate increased infection-associated mortality and bacterial burdens. Unexpectedly, numbers of Salmonella-specific CD4+ T cells were significantly increased in thymectomized mice compared to sham control mice. Lastly, we found that T cells from thymectomized mice may be impaired in producing the effector cytokine IL-17 at early time points of infection, compared to thymically intact mice. Together, these results imply a unique role for thymic output in the formation of immune responses against a persistent, enteric pathogen.

Aquatic thermal conditions predict the presence of native and invasive rock pool Aedes (Diptera: Culicidae) in the southern Appalachians, U.S.A.

The native rock pool mosquito, Aedes atropalpus (Coquillett), and the invasive Aedes japonicus (Theobald) have been found in many types of artificial and natural containers throughout North America. Little is known about the ecology of these two species in habitats where they co-occur, although multiple investigators have reported the decline of the native species concurrent with the introduction and spread of the invasive species. Here we report the results of riverine rock pool collections (n=503) in the southern Appalachian Mountains between 2009-2015. Surface water temperatures strongly predicted the presence of each species across a broad range of observed temperatures (11-39.8° C). For every unit of increase in temperature (°C) the odds of collecting Ae. atropalpus larvae increased by 0.34 while the odds of collecting Ae. japonicus larvae decreased by 0.28. No Ae. japonicus larvae or pupae were collected at temperatures greater than 36° C; however, immature Ae. atropalpus were found in rock pools with temperatures up to 39.8° C. In contrast, Ae. japonicus were highly abundant in cooler rock pools (<17° C) where Ae. atropalpus were infrequent or absent. Our findings suggest that in spite of the successful invasion by Ae. japonicus, Ae. atropalpus remains well established in the southern Appalachian Mountains. Given the strong correlation of temperature with the presence of the two species and the contrasting absence of each species at observed temperature extremes, the role of thermal conditions should be carefully explored in the context of other ecological factors likely influencing the range and abundance of these mosquitoes.

Adjuvant selection regulates gut migration and phenotypic diversity of antigen-specific CD4+ T cells following parenteral immunization.

Infectious diarrheal diseases are the second leading cause of death in children under 5 years, making vaccines against these diseases a high priority. It is known that certain vaccine adjuvants, chiefly bacterial ADP-ribosylating enterotoxins, can induce mucosal antibodies when delivered parenterally. Based on this, we reasoned vaccine-specific mucosal cellular immunity could be induced via parenteral immunization with these adjuvants. Here, we show that, in contrast to the Toll-like receptor-9 agonist CpG, intradermal immunization with non-toxic double-mutant heat-labile toxin (dmLT) from enterotoxigenic Escherichia coli drove endogenous, antigen-specific CD4+ T cells to expand and upregulate the gut-homing integrin α4ß7. This was followed by T-cell migration into gut-draining lymph nodes and both small and large intestines. We also found that dmLT produces a balanced T-helper 1 and 17 (Th1 and Th17) response, whereas T cells from CpG immunized mice were predominantly Th1. Immunization with dmLT preferentially engaged CD103+ dendritic cells (DCs) compared with CpG, and mice deficient in CD103+ DCs were unable to fully license antigen-specific T-cell migration to the intestinal mucosae following parenteral immunization. This work has the potential to redirect the design of existing and next generation vaccines to elicit pathogen-specific immunity in the intestinal tract with non-mucosal immunization.

Salmonella infection: Interplay between the bacteria and host immune system.

Salmonella infection causes morbidity and mortality throughout the world with the host immune response varying depending on whether the infection is acute and limited, or systemic and chronic. Additionally, Salmonella bacteria have evolved multiple mechanisms to avoid or subvert immunity to its own benefit and often the anatomical location of infection plays a role in both the immune response and bacterial fate. Here, we provide an overview of the interplay between the immune system and Salmonella, while discussing how different host and bacterial factors influence the outcome of infection.

Synovitis and the risk of knee osteoarthritis: the MOST Study.

OBJECTIVE: To identify the independent relation of synovitis with incident radiographic knee osteoarthritis (OA) after adjusting for other structural factors known to cause synovitis. DESIGN: We examined MRIs from knees that developed incident radiographic OA from the Multicenter Osteoarthritis Study (MOST) and compared these case knees with controls that did not develop OA. We examined baseline MRIs for knees developing OA at any time up to 84 months follow-up. We scored lesions in cartilage, meniscus, bone marrow and synovitis. Synovitis scores were summed (0-9) across three regions, suprapatellar, infrapatellar and intercondylar region, each of which was scored 0-3. After bivariate analyses examining each factor's association with incidence, we carried out multivariable regression analyses adjusting for age, sex, BMI, alignment and cartilage and meniscal damage. RESULTS: We studied 239 case and 731 control knees. In bivariate analyses, cartilage lesions, meniscal damage, synovitis and bone marrow lesions were all risk factors for OA. After multivariable analyses, synovitis was associated with incident OA. A higher synovitis score increased the risk of incident OA (adjusted OR per unit increase 1.1; (95% CI 1.0, 1.2, P = .02)), but increased risk was associated only with synovitis scores of ≥3 (adjusted OR 1.6; 95% CI 1.2, 2.1, P = .003). CONCLUSIONS: Synovitis, especially when there is a substantial volume within the knee, is an independent cause of OA.

Physical activity, alignment and knee osteoarthritis: data from MOST and the OAI.

OBJECTIVE: To determine the effect of physical activity on knee osteoarthritis (OA) development in persons without knee injury and according to knee alignment. DESIGN: We combined data from Multicenter Osteoarthritis (MOST) and Osteoarthritis Initiative (OAI), studies of persons with or at high risk of OA. Subjects had long limb and repeated posteroanterior knee radiographs and completed the physical activity survey for the elderly (PASE). We studied persons without radiographic OA and excluded knees with major injury and without long limb films. We followed subjects 30 months (in MOST) and 48 months (in OAI) for one of two incident outcomes: (1) symptomatic tibiofemoral OA (radiographic OA and knee pain), or (2) tibiofemoral narrowing. 'Active' persons were those with PASE score in the highest quartile by gender. We examined risk of OA in active group using logistic regression adjusting for age, gender, body mass index (BMI), Western Ontario and McMaster Arthritis Index (WOMAC) pain score, Kellgren and Lawrence (KL) grade (0 or 1), and study of origin. We also analyzed knees from malaligned and neutrally aligned limbs. RESULTS: The combined sample comprised 2,073 subjects (3,542 knees) with mean age 61 years. The cumulative incidence of symptomatic tibiofemoral OA was 1.12% in the active group vs 1.82% in the others (odds ratio (OR) among active group 0.6, 95% confidence interval (CI) 0.3, 1.3). Joint space narrowing occurred in 3.41% of knees in the active group vs 4.04% in the others (OR among active group 0.9 (95% CI 0.5, 1.5)). Results did not differ by alignment status. CONCLUSIONS: Physical activity in the highest quartile did not affect the risk of developing OA.

Can anatomic alignment measured from a knee radiograph substitute for mechanical alignment from full limb films?

OBJECTIVES: To examine whether categories of anatomic alignment (varus, neutral, valgus) measured from knee X-rays agree with similar categories of mechanical alignment from the full limb film and whether varus anatomic malalignment predicts medial joint space loss on knee X-rays as well as varus mechanical alignment. METHODS: We used data from the Osteoarthritis Initiative (OAI) (full limb and flexed knee X-rays) to examine agreement of anatomic and mechanical alignment and data from Boston Osteoarthritis of the Knee Study (BOKS) to evaluate the association of full limb mechanical alignment vs knee X-ray anatomic alignment with joint space loss. A 4 degree offset was used to correct for the more valgus angulation of the anatomic alignment. RESULTS: Of 143 subjects whose knee X-rays and full limb films were publicly released from the OAI, the agreement of varus, neutral and valgus alignment was only moderate (kappa=0.43, P<0.001). In BOKS, varus mechanical and anatomic alignments measured from full limb and knee X-rays respectively both predicted a high risk of medial joint space loss vs neutral alignment--for mechanical alignment, odds ratio (OR)=4.82 [95% confidence interval (CI) 1.93, 12.00] and for anatomic alignment OR=4.25 (95% CI 2.08, 8.72). CONCLUSIONS: While agreement of alignment from knee X-ray to full limb film was only moderate, varus malalignment measured from a flexed knee predicted the likelihood of progression well. Flexed knee alignment may be more relevant to knee osteoarthritis (OA) risk than that of a fully extended knee, but a measurement of alignment from a short limb is an imperfect surrogate for full limb alignment.

The association between patellar alignment and patellofemoral joint osteoarthritis features--an MRI study.

OBJECTIVE: The aim of our study was to evaluate the association between patellar alignment (using standard MRI images of extended knees) and MRI indices of patellofemoral (PF) osteoarthritis (OA) features. METHODS: In this cross-sectional observational study, subjects were recruited to participate in the Boston Osteoarthritis of the Knee Study (BOKS). The association of patellar alignment [patellar length ratio (PLR), sulcus angle (SA), lateral patellar tilt angle (LPTA) and bisect offset (BO)] with measures of PF OA [cartilage morphology and bone marrow lesion (BML) in the medial and lateral PF compartment] were examined using a logistic regression model while adjusting for age, sex and BMI. RESULTS: Study sample comprised 126 males (mean age 68.0, BMI 31.2) and 87 females (mean age 64.7, BMI 31.6). All measurements of patellar alignment were statistically significantly associated with cartilage morphology and BML in the lateral compartment of PF joint. PLR and SA were significantly associated with medial cartilage loss. With increasing PLR there was an increased prevalence of lateral and medial cartilage loss as well as of lateral BML. Increasing SA was positively associated with increased lateral and medial cartilage loss and lateral BML. LPTA range was negatively associated with lateral cartilage loss and BML. More laterally displaced patella (higher BO) was associated with increased lateral cartilage loss and BML. CONCLUSIONS: The results of our study clearly indicated that patellar alignment is associated with manifestations of PF OA such as cartilage thickness loss and BML.

Osteophytes and progression of knee osteoarthritis.

OBJECTIVES: Osteophytes are thought to stabilize an osteoarthritic joint, thereby preventing structural progression. Meagre longitudinal data suggest, however, that they are associated with an increased risk of structural progression. Our objective was to evaluate the effect of osteophyte size on radiographic progression in osteoarthritis (OA). METHODS: Using data from a natural history study of persons with symptomatic knee OA, we obtained fluoroscopically positioned postero-anterior (PA) radiographs at baseline, 15 and 30 months. Using an atlas, osteophyte size was scored on a scale of 0-3 at each of four sites on the PA film and, for each knee, both compartment-specific (i.e. medial; lateral) and overall osteophyte scores were computed. Progression was defined as an increase over follow-up in medial or lateral joint space narrowing, based on a semiquantitative grading. Mechanical alignment was assessed using long limb films at the 15 month examination. Logistic regression was used to evaluate the relation of osteophyte size with progression, adjusting for age, gender and body mass index, and with and without adjustment for alignment. RESULTS: Of 270 subjects who had 470 eligible knees with follow-up, 104 (22%) knees showed progression. Overall, osteophyte score modestly increased the risk of progression [odds ratio (OR) per S.D. increase of osteophyte score=1.4 (95% CI 1.1, 1.8, P=0.02)], but this effect weakened and became non-significant after adjustment for limb alignment (OR=1.3). Compartment osteophyte score was strongly associated with malalignment to the side of the osteophyte (e.g. medial osteophyte and varus). Compartment-specific osteophyte score markedly increased the risk of ipsilateral progression (e.g. medial osteophytes --> medial progression) [OR per S.D.=1.9 (95% CI 1.5, 2.5, P<0.001)] and decreased the risk of contralateral progression [OR per S.D.= 0.6 (95% CI 0.5, 0.8, P=0.002)], but these associations diminished when we adjusted for limb alignment (OR=1.5 and 0.7 respectively). CONCLUSIONS: Large osteophytes do not affect the risk of structural progression. They are strongly associated with malalignment to the side of the osteophyte, and any relation they have with progression is partly explained by the association of malalignment with progression.

Relationship between arterial thrombosis and neutralization of a polyethylene ionomer.

The influence of three levels of sodium neutralization of an ethylene/methacrylic acid copolymer on in vivo blood compatibility was studied in a canine arterial model. Effects due to sample fabrication methods (thermal pressing versus solution casting) were also monitored. Sodium content, sodium release, hydrogen dissociation, and localization of anionic groups were noted. Polymer surface energy, surface morphology, water uptake, and thermal properties were characterized. Material characterization and in vivo implantation disclose the following: 1) Thermal pressing generated oxidation degradation products that decreased in vivo blood compatibility. Solution-cast samples adhered and activated fewer blood elements. 2) Platelets and leukocytes were sensitive to differences in shear rate in the carotid and femoral arteries, with the femoral site tending toward higher shear, more platelet deposition and fewer leukocytes. 3) The surface properties of the polyethylene control, 0% Na, and 50% Na samples tended to be similar. These properties were different from the 100% Na sample. The 100% Na ionomer was more hydrophilic, had a higher polar component for its surface energy, and was unique in exhibiting discrete ionic clusters 1-10 microns in diameter on its surface. 4) These differences were manifested in vivo by platelet activation and thrombus development on the polyethylene, 0% Na, and 50% Na implants, while the 100% Na implant surfaces were predominantly covered by singly adherent, unactivated platelets. 5) It is proposed that the improvement in biocompatibility for the 100% Na ionomer is due to the cluster development in the neutralized methacrylic component and that either directly, or through appropriate protein adsorption and/or conformational adjustment to the cluster regions, platelets are not activated and do not initiate the coagulation mechanism.

Vascular graft sealants.

Although the use of vascular graft sealants is primarily intended to decrease the porosity of grafts upon implantation, sealants also may serve to passivate polymeric surfaces or act as a temporary scaffold for cell attachment and subsequent graft healing. The safety and efficacy of vascular grafts sealed with albumin, collagen, and gelatin have been established through many preclinical and clinical studies. Such products have been used in clinical practice since the mid-1980s. Any differences in the long-term human healing response to these sealants and the clinical relevance of such differences can be assessed only as the time of implantation increases and the data are evaluated.

Tissue culture of fetal rat islets: corticosterone promotes D cell maintenance and function.

Fetal pancreatic islets were cultured using a recently described technique (1). After 1 day in culture, half of the plates were continued in control medium and half were grown in identical medium supplemented with corticosterone (0.1 microgram/ml). Media were renewed daily, and culture was continued for a total of 8 days. Insulin, glucagon, and somatostatin contents in the media were determined daily. These hormones were also estimated in the tissue at the time of plating, and after 1 and 8 days in culture. Islets were fixed on day 8, and the cells containing each of these hormones were identified by immunocytochemical staining. Corticosterone supplementation of the medium resulted in a 3-fold increase in the somatostatin concentration of the medium. The insulin and glucagon contents of the supplemented medium were slightly reduced. On day 8, there was no difference in the insulin content of the cultured tissue regardless of medium. The glucagon and somatostatin contents of the tissue grown in the steroid-supplemented medium were greater (1.8- and 3.1-fold, respectively) than those of the tissue grown in control medium. D cells were rare in the islets grown in control medium volume density, 0.4%, but were more numerous in the islets maintained in supplemented medium (2.2%). Islets grown in corticosterone-supplemented medium lacked an insulin secretory response to 22 mM glucose. These findings indicate that the volume densities of the cells within the islets can be altered during an 8-day period in culture, suggesting that nutritional and other requirements of the individual subpopulations of islet cells may be different. In addition, corticosterone may prevent the maturation of the secretory responsiveness of cultured B cells to glucose.

Research and development contracting at the National Institute of Dental Research.

Government contracting for biomedical research and development is a cooperative venture of high public visibility. It must be soundly conceived, openly instituted, and skillfully managed. Because of the cooperative nature of contract activities, it is important for the scientific community to understand the contract mechanism and the concepts upon which its use is based. Therefore, these concepts are described and how this mechanism serves as an aid in accomplishing the mission of the National Institute of Dental Research is discussed.