Development of novel multi-protein chimeric immunogens that protect against infection with the Lyme disease agent, Borreliella burgdorferi.

Lyme disease is the most common tick-borne disease in North America. A vaccine for use in humans is not available. Here, we detail the development of two chimeric vaccine antigens, BAF and Chv2M. BAF elicits Abs that target proteins and protein variants produced by Borreliella species in ticks (OspB and OspA) and mammals (FtlA/B). OspB serves as the backbone structure for the BAF chimeric. Two OspA221-240 epitope-containing domain (ECD) variants (#A1 and #A15) were inserted into a loop in OspB. The N-terminal region of the FtlA protein was joined to the C-terminus of the chimeric. The second chimeric, Chv2M, consists of L5 (loop 5) and H5 (helix 5) ECDs derived from diverse OspC proteins. Borreliella species produce OspC upon exposure to the bloodmeal and during early infection in mammals. Here, we demonstrate that BAF and Chv2M are potent immunogens that elicit Abs that bind to each component protein (FtlA, FtlB, OspB, and multiple OspA and OspC variants). Anti-BAF and anti-Chv2M Abs kill Borreliella burgdorferi strains through Ab-mediated complement-dependent and complement-independent mechanisms. Eighty percent (32/40) of mice that received a three-dose vaccine regimen were protected from infection with B. burgdorferi B31. Efficacy increased to 90% (18/20) when the amount of Chv2M was increased in the third vaccine dose. Readouts for infection were flaB PCR and seroconversion to VlsE. This study establishes proof of principle for a chimeric immunogen vaccine formulation that elicits Abs to multiple targets on the B. burgdorferi cell surface produced during tick and mammalian stages of the enzootic cycle.IMPORTANCELyme disease is a growing public health threat across parts of the Northern Hemisphere. Regions that can support sustained tick populations are expanding, and the incidence of tick-borne diseases is increasing. In light of the increasing risk of Lyme disease, effective preventive strategies are needed. Most vaccine development efforts have focused on outer surface protein A, a Borreliella burgdorferi protein produced only in ticks. Herein, we describe the development of a novel vaccine formulation consisting of two multivalent chimeric proteins that are immunogenic and elicit antibodies with bactericidal activity that target several cell surface proteins produced by the Lyme disease spirochetes in feeding ticks and mammals. In a broader sense, this study advances efforts to develop custom-designed vaccinogens comprised of epitope-containing domains from multiple proteins.

Rationale and design of the Baylor Infant Twin Study-A study assessing obesity-related risk factors from infancy.

BACKGROUND: Early childhood (0-3 years) is a critical period for obesity prevention, when tendencies in eating behaviors and physical activity are established. Yet, little is understood about how the environment shapes children's genetic predisposition for these behaviors during this time. The Baylor Infant Twin Study (BITS) is a two phase study, initiated to study obesity risk factors from infancy. Data collection has been completed for Phase 1 in which three sub-studies pilot central measures for Phase 2. A novel infant temperament assessment, based on observations made by trained researchers was piloted in Behavior Observation Pilot Protocol (BOPP) study, a new device for measuring infant feeding parameters (the "orometer") in the Baylor Infant Orometer (BIO), and methods for analyzing DNA methylation in twins of unknown chorionicity in EpiTwin. METHODS: EpiTwin was a cross-sectional study of neonatal twins, while up to three study visits occurred for the other studies, at 4- (BOPP, BIO), 6- (BOPP), and 12- (BOPP, BIO) of age. Measurements for BOPP and BIO included temperament observations, feeding observations, and body composition assessments while EpiTwin focused on collecting samples of hair, urine, nails, and blood for quantifying methylation levels at 10 metastable epialleles. Additional data collected include demographic information, zygosity, chorionicity, and questionnaire-based measures of infant behaviors. RESULTS: Recruitment for all three studies was completed in early 2020. EpiTwin recruited 80 twin pairs (50% monochorionic), 31 twin pairs completed the BOPP protocol, and 68 singleton infants participated in BIO. CONCLUSIONS: The psychometric properties of the data from all three studies are being analyzed currently. The resulting findings will inform the development of the full BITS protocol, with the goal of completing assessments at 4-, 6-, 12-, and 14-month of age for 400 twin pairs.

A genomic atlas of systemic interindividual epigenetic variation in humans.

BACKGROUND: DNA methylation is thought to be an important determinant of human phenotypic variation, but its inherent cell type specificity has impeded progress on this question. At exceptional genomic regions, interindividual variation in DNA methylation occurs systemically. Like genetic variants, systemic interindividual epigenetic variants are stable, can influence phenotype, and can be assessed in any easily biopsiable DNA sample. We describe an unbiased screen for human genomic regions at which interindividual variation in DNA methylation is not tissue-specific. RESULTS: For each of 10 donors from the NIH Genotype-Tissue Expression (GTEx) program, CpG methylation is measured by deep whole-genome bisulfite sequencing of genomic DNA from tissues representing the three germ layer lineages: thyroid (endoderm), heart (mesoderm), and brain (ectoderm). We develop a computational algorithm to identify genomic regions at which interindividual variation in DNA methylation is consistent across all three lineages. This approach identifies 9926 correlated regions of systemic interindividual variation (CoRSIVs). These regions, comprising just 0.1% of the human genome, are inter-correlated over long genomic distances, associated with transposable elements and subtelomeric regions, conserved across diverse human ethnic groups, sensitive to periconceptional environment, and associated with genes implicated in a broad range of human disorders and phenotypes. CoRSIV methylation in one tissue can predict expression of associated genes in other tissues. CONCLUSIONS: In addition to charting a previously unexplored molecular level of human individuality, this atlas of human CoRSIVs provides a resource for future population-based investigations into how interindividual epigenetic variation modulates risk of disease.

The Female Genital Self-image Scale in Adult Women With Vulvar Lichen Sclerosus.

OBJECTIVE: There is no agreed upon standard way to measure vulvar lichen sclerosus disease severity. The Female Genital Self-Image Scale (FGSIS) is a validated survey tool assessing female genital self-image and is positively correlated with women's sexual function. A lower score represents a negative genital self-image. We evaluated the FGSIS in women with vulvar lichen sclerosus. METHODS: Women with biopsy-proven lichen sclerosus and women presenting for routine gynecologic care without lichen sclerosus matched by age were surveyed with the 7-item FGSIS. National surveys of healthy women in the United States have shown the mean 7-item FGSIS score is 21. To detect one standard deviation (20% absolute difference) between groups with a power of 80% at p < .05, 15 women would are needed in each group. RESULTS: Sixteen women with lichen sclerosus and 16 matched controls were surveyed between February and July 2018. The mean ± SD age of women with lichen sclerosus was 56.8 ± 13.5 years, 94% were white, 69% married, 81% college educated, 69% postmenopausal, and 18% on hormone replacement therapy. None of these differences were statistically different from control women. Women with vulvar lichen sclerosus had a significantly lower median 7-item FGSIS when compared with control subjects, 18 (interquartile range = 16-21) versus 25 (interquartile range = 23-27), respectively, Mann-Whitney U test, p < .001. CONCLUSIONS: Women with vulvar lichen sclerosus have a lower score on the 7-item Female Genital Self-Image Scale compared with healthy controls.

Prevention and management of hysterectomy complications.

Hysterectomy is the most common gynecologic surgical procedure performed in the United States. Although most hysterectomies proceed without incident, complications with serious consequences may occur. This chapter reviews the incidence, predisposing factors, intraoperative risk, diagnosis, and management and prevention of complications of hysterectomy. These include hemorrhage, infection, thromboembolism, injury to viscera, and neuropathy. The prepared surgeon is familiar with anatomy, surgical risk factors, current recommendations for prophylaxis and prevention, as well as modern management of complications of hysterectomy.

Hysteroscopic sterilization in the office setting.

Hysteroscopic sterilization is growing in popularity. Nearly 500,000 women have been sterilized using this method, and an increasing number of physicians are now performing this procedure in the office setting. The office setting can provide a cost-effective, convenient, and safe environment for hysteroscopic sterilization. Patients may benefit from avoiding hospital preoperative visits, excessive laboratory evaluation, operating room wait times, and expense associated with hospital care. Physicians may improve productivity through remaining in their office or avoiding operating room delays. This article reviews office-hysteroscopic sterilization with the Essure microinsert system.

Salivary progesterone levels and rate of ovulation are significantly lower in poorer than in better-off urban-dwelling Bolivian women.

BACKGROUND: Agriculturalists in less-developed countries (LDC) have lower progesterone levels than urban industrialized populations. However, it is unknown if urban LDC populations are also relatively lower. We tested whether urban Bolivia samples-poorer (Bol-p) and better-off (Bol-b)-have lower progesterone than a Chicago (USA) sample, and whether progesterone and rate of ovulation are lower in Bol-p than in Bol-b. METHODS: Serial salivary samples collected from Bolivians, screened according to strict exclusion criteria during two complete menstrual cycles, were radioimmunoassayed for progesterone; anthropometrics were collected at mid-follicular and mid-luteal phases. RESULTS: Progesterone levels are lower in the Bolivia samples, and higher in the Bol-b than Bol-p; ovulation rate is greater in Bol-b than Bol-p. For only ovulatory cycles, mean-follicular-P (pmol/l), mean-luteal-P (pmol/l), and mean-peak-P (pmol/l) are respectively 65, 142 and 208 in Bol-p; 76, 167 and 232 in Bol-b; and 96, 240 and 330 in Chicago. Principal components representing body-size and progesterone level are positively correlated (r = 0.404, P = 0.005). CONCLUSIONS: Progesterone levels appear to be influenced by chronic and acute ecological conditions, evidenced by the association with body-size and the probability of ovulation respectively. These findings have implications for understanding cancer aetiology, developing population-appropriate hormonal contraceptives, and modelling the evolution and functioning of the reproductive system.