Sex Hormones Regulate Tenofovir-Diphosphate in Female Reproductive Tract Cells in Culture

The conflicting results of recent pre-exposure prophylaxis (PrEP) trials utilizing tenofovir (TFV) to prevent HIV infection in women led us to evaluate the accumulation of intracellular TFV-diphosphate (TFV-DP) in cells from the female reproductive tract (FRT) and whether sex hormones influence the presence of TFV-DP in these cells. Following incubation with TFV, isolated epithelial cells, fibroblasts, CD4+ T cells and CD14+ cells from the FRT as well as blood CD4+ T cells and monocyte-derived macrophages convert TFV to TFV-DP. Unexpectedly, we found that TFV-DP concentrations (fmol/million cells) vary significantly with the cell type analyzed and the site within the FRT. Epithelial cells had 5-fold higher TFV-DP concentrations than fibroblasts; endometrial epithelial cells had higher TFV-DP concentrations than cells from the ectocervix. Epithelial cells had 125-fold higher TFV-DP concentrations than FRT CD4+ T cells, which were comparable to that measured in peripheral blood CD4+ T cells. These findings suggest the existence of a TFV-DP gradient in the FRT where epithelial cells \u3e fibroblasts \u3e CD4+ T cells and macrophages. In other studies, estradiol increased TFV-DP concentrations in endometrial and endocervical/ectocervical epithelial cells, but had no effect on fibroblasts or CD4+ T cells from FRT tissues. In contrast, progesterone alone and in combination with estradiol decreased TFV-DP concentrations in FRT CD4+ T cells. Our results suggest that epithelial cells and fibroblasts are a repository of TFV-DP that is under hormonal control. These cells might act either as a sink to decrease TFV availability to CD4+ T cells and macrophages in the FRT, or upon conversion of TFV-DP to TFV increase TFV availability to HIV-target cells. In summary, these results indicate that intracellular TFV-DP varies with cell type and location in the FRT and demonstrate that estradiol and/or progesterone regulate the intracellular concentrations of TFV-DP in FRT epithelial cells and CD4+ T cells

Seasonal changes in anthropometric and physical characteristics within English academy rugby league players.

Professional rugby league clubs implement training programmes for the development of anthropometric and physical characteristics within an academy programme. However, research that examines seasonal changes in these characteristics is limited. The purpose of the study was to evaluate the seasonal changes in anthropometric and physical characteristics of academy rugby league players by age category (i.e., under 14, 16, 18, 20). Data were collected on 75 players pre- and postseason over a 6-year period (resulting in a total of 195 assessments). Anthropometric (body mass, sum of 4 skinfolds) and physical (10- and 20-m sprint, vertical jump, Yo-Yo intermittent recovery test and 1 repetition maximum squat, bench press, and prone row) measures were collected. The under 14s and 16s showed greater seasonal improvements in body mass (e.g., under 14s = 7.4 ± 4.3% vs. under 20s = 1.2 ± 3.3%) and vertical jump performance than under 18s and under 20s. In contrast, under 18s and under 20s players showed greater seasonal improvements in Yo-Yo performance and 10-m sprint (e.g., under 14s = 1.3 ± 3.9% vs. under 20s = -1.9 ± 1.2%) in comparison to under 14s and under 16s. Seasonal strength improvements were greater for the under 18s compared with under 20s. This study provides comparative data for seasonal changes in anthropometric and physical characteristics within rugby league players aged 13-20 years. Coaches should be aware that seasonal improvements in speed may not exist within younger age categories, until changes in body mass stabilize and consider monitoring changes in other characteristics (e.g., momentum). Large interplayer variability suggests that player development should be considered on an individual and longitudinal basis

Peak Running Speeds in Professional Male Football: Influence of Division and Playing Position

Well-established physical demands of competitive professional football facilitate prescription and monitoring of training. However, many factors influence these physical demands with implications for efficacious practice. Match-play data were analyzed over 2 seasons using global positioning systems technology, differentiating English Championship (33 matches) and League One (27 matches) demands. Playing position categorized wide and central defenders and midfielders and forwards. Peak running speeds defined the outcome measure, assessing the influence of the competition level and playing position across 1, 5, and 10-minute rolling average durations using a linear mixed model. Significant effects were detected for the competition level (F 1,324.5 = 5.44, p = 0.02) and playing position (F 4,328.3 = 89.90, p < 0.001). League One matches demonstrated greater peak running speeds than Championship matches (mean difference = 2.72 m·min−1 [95% confidence intervals: 0.4, 5.0]). No difference was observed between central and wide midfielders (mean difference = 0.62 m·min−1 [95% confidence intervals: −3.1, 4.3]). Wide midfielders presented faster peak running speeds than forwards (mean difference = 18 m·min−1 [95% confidence intervals:14.1, 22.1], p < 0.05), central defenders (mean difference = 25 m·min−1 [95% confidence intervals: 21.7, 29.8], p < 0.05), and wide defenders (mean difference = 12 m·min−1 [95% confidence intervals: 8.2, 16.5], p < 0.05). Interaction effects were found for division*position (F 4,328.3 = 2.57, p = 0.038) demonstrating greater running speeds in League One, except for central defenders. Wide midfielders presented greater peak 1-minute running speeds, whereas 5 and 10-minute peak running speeds were greatest in central midfielders. The sensitivity of peak running speeds to competition level and playing position has implications for training prescription, monitoring particularly when transitioning between competition levels, determining and monitoring positional training intensities, and objective targets for progressive overload during rehabilitation

Anthropometric and physical characteristics of english academy rugby league players.

The purpose of the present study was to evaluate the anthropometric and physical characteristics of English academy rugby league players by annual-age category (under 16s-under 20s) and between backs and forwards. Data were collected on 133 academy players over a 6-year period (resulting in a total of 257 assessments). Player assessments comprised of anthropometric (height, body mass, sum of 4 skinfolds) and physical (vertical jump, 10- and 20-m sprint, estimated V[Combining Dot Above]O2max via the yo-yo intermittent recovery test level 1, absolute 1 repetition maximum [1RM], and relative squat, bench press, and prone row) measures. Univariate analysis of variance demonstrated significant (p ≤ 0.05) increases in height, body mass, vertical jump, absolute, and relative strength measures across the 5 annual-age categories (e.g., body mass: under 16s = 75.2 ± 11.1, under 20s = 88.9 ± 8.5 kg; vertical jump: under 16s = 45.7 ± 5.2, under 20s = 52.8 ± 5.4 cm; 1RM bench press: under 16s = 73.9 ± 13.2, under 20s = 114.3 ± 15.3 kg). Independent t-tests identified significant (p ≤ 0.05) differences between backs and forwards for anthropometric (e.g., under 16s body mass: backs = 68.4 ± 8.6, forwards = 80.9 ± 9.7 kg) and physical (e.g., under 19s 20-m sprint: backs = 3.04 ± 0.08, forwards = 3.14 ± 0.12s; under 18s relative squat: backs = 1.65 ± 0.18, forwards = 1.51 ± 0.17 kg·kg) characteristics that were dependent on the age category and measure assessed. Findings highlight that anthropometric and physical characteristics develop across annual-age categories and between backs and forwards in academy rugby league players. These findings provide comparative data for such populations and support the need to monitor player development in junior rugby league players

Estradiol Regulation of Nucleotidases in Female Reproductive Tract Epithelial Cells and Fibroblasts

The use of topical and oral adenosine derivatives in HIV prevention that need to be maintained in tissues and cells at effective levels to prevent transmission prompted us to ask whether estradiol could influence the regulation of catabolic nucleotidase enzymes in epithelial cells and fibroblasts from the upper and lower female reproductive tract (FRT) as these might affect cellular TFV-DP levels. Epithelial cells and fibroblasts were isolated from endometrium (EM), endocervix (CX) and ectocervix (ECX) tissues from hysterectomy patients, grown to confluence and treated with or without estradiol prior to RNA isolation. The expression of nucleotidase (NT) genes was measurable by RT-PCR in epithelial cells and fibroblasts from all FRT tissues. To determine if sex hormones have the potential to regulate NT, we evaluated NT gene expression and NT biological activity in FRT cells following hormone treatment. Estradiol increased expression of Cytosolic 5 9 -nucleotidase after 2 or 4 h in endometrial epithelial cells but not epithelial cells or fibroblasts from other sites. In studies using a modified 5 9 - Nucleotidase biological assay for nucleotidases, estradiol increased NT activity in epithelial cells and fibroblasts from the EM, CX and ECX at 24 and 48 h. In related studies, HUVEC primary cells and a HUVEC cell line were unresponsive to estradiol in terms of nucleotidase expression or biological activity. Our findings of an increase in nucleotidase expression and biological activity induced by estradiol do not directly assess changes in microbicide metabolism. However, they do suggest that when estradiol levels are elevated during the menstrual cycle, FRT epithelial cells and fibroblasts from the EM, CX and ECX have the potential to influence microbicide levels that could enhance protection of HIV-target cells (CD4 + T cells, macrophages and dendritic cells) throughout the FRT

Structure Preserving Parallel Algorithms for Solving the Bethe-Salpeter Eigenvalue Problem

The Bethe-Salpeter eigenvalue problem is a dense structured eigenvalue problem arising from discretized Bethe-Salpeter equation in the context of computing exciton energies and states. A computational challenge is that at least half of the eigenvalues and the associated eigenvectors are desired in practice. We establish the equivalence between Bethe-Salpeter eigenvalue problems and real Hamiltonian eigenvalue problems. Based on theoretical analysis, structure preserving algorithms for a class of Bethe-Salpeter eigenvalue problems are proposed. We also show that for this class of problems all eigenvalues obtained from the Tamm-Dancoff approximation are overestimated. In order to solve large scale problems of practical interest, we discuss parallel implementations of our algorithms targeting distributed memory systems. Several numerical examples are presented to demonstrate the efficiency and accuracy of our algorithms

Validation of the Aura Microwave Limb Sounder HNOmeasurements

We assess the quality of the version 2.2 (v2.2) HNO3 measurements from the Microwave Limb Sounder (MLS) on the Earth Observing System Aura satellite. The MLS HNO3 product has been greatly improved over that in the previous version (v1.5), with smoother profiles, much more realistic behavior at the lowest retrieval levels, and correction of a high bias caused by an error in one of the spectroscopy files used in v1.5 processing. The v2.2 HNO3 data are scientifically useful over the range 215 to 3.2 hPa, with single-profile precision of ∼0.7 ppbv throughout. Vertical resolution is 3–4 km in the upper troposphere and lower stratosphere, degrading to ∼5 km in the middle and upper stratosphere. The impact of various sources of systematic uncertainty has been quantified through a comprehensive set of retrieval simulations. In aggregate, systematic uncertainties are estimated to induce in the v2.2 HNO3 measurements biases that vary with altitude between ±0.5 and ±2 ppbv and multiplicative errors of ±5–15% throughout the stratosphere, rising to ∼±30% at 215 hPa. Consistent with this uncertainty analysis, comparisons with correlative data sets show that relative to HNO3 measurements from ground-based, balloon-borne, and satellite instruments operating in both the infrared and microwave regions of the spectrum, MLS v2.2 HNO3 mixing ratios are uniformly low by 10–30% throughout most of the stratosphere. Comparisons with in situ measurements made from the DC-8 and WB-57 aircraft in the upper troposphere and lowermost stratosphere indicate that the MLS HNO3 values are low in this region as well, but are useful for scientific studies (with appropriate averaging)

The Kneeling Isometric Plantar Flexor Test: Preliminary Reliability and Feasibility in Professional Youth Football

Calf injuries are common in professional football; thus, the establishment of reliable and time-efficient methods of measuring the peak force capabilities of the plantar flexors with equipment that is accessible to football practitioners is valuable. In this study, we determined the preliminary reliability and feasibility of a new test, termed the kneeling isometric plantar flexion test (KIPFT), for footballers. Twenty-one male youth footballers (age = 17.8 ± 1.1 years, height = 182 ± 5 cm, weight = 77.6 ± 5.9 kg) from English League One football clubs completed three trials of the KIPFT on a wireless force plate at the end (2022–2023) and start (2023–2024) of the season. The within-session reliability of the peak force (relative to body weight) was good–excellent for both limbs and both occasions. On average, performance of the KIPFT took just over 1 min per limb and ~2 min to set up. The peak force values were larger for the non-dominant limbs only at the start versus the end of the season, but there were no between-limb differences. From these results, it was determined that (1) the KIPFT is feasible, (2) a minimum of 32 footballers would be required to establish its between-session reliability with ≥80% statistical power and (3) large-cohort normative data for the KIPFT may be best collected at the start of the football season

A Phase 1 Trial of MSP2-C1, a Blood-Stage Malaria Vaccine Containing 2 Isoforms of MSP2 Formulated with Montanide® ISA 720

Background: In a previous Phase 1/2b malaria vaccine trial testing the 3D7 isoform of the malaria vaccine candidate Merozoite surface protein 2 (MSP2), parasite densities in children were reduced by 62%. However, breakthrough parasitemias were disproportionately of the alternate dimorphic form of MSP2, the FC27 genotype. We therefore undertook a dose-escalating, double-blinded, placebo-controlled Phase 1 trial in healthy, malaria-naïve adults of MSP2-C1, a vaccine containing recombinant forms of the two families of msp2 alleles, 3D7 and FC27 (EcMSP2-3D7 and EcMSP2-FC27), formulated in equal amounts with Montanide® ISA 720 as a water-in-oil emulsion. Methodology/Principal Findings: The trial was designed to include three dose cohorts (10, 40, and 80 μg), each with twelve subjects receiving the vaccine and three control subjects receiving Montanide® ISA 720 adjuvant emulsion alone, in a schedule of three doses at 12-week intervals. Due to unexpected local reactogenicity and concern regarding vaccine stability, the trial was terminated after the second immunisation of the cohort receiving the 40 μg dose; no subjects received the 80 μg dose. Immunization induced significant IgG responses to both isoforms of MSP2 in the 10 μg and 40 μg dose cohorts, with antibody levels by ELISA higher in the 40 μg cohort. Vaccine-induced antibodies recognised native protein by Western blots of parasite protein extracts and by immunofluorescence microscopy. Although the induced anti-MSP2 antibodies did not directly inhibit parasite growth in vitro, IgG from the majority of individuals tested caused significant antibody-dependent cellular inhibition (ADCI) of parasite growth. Conclusions/Significance: As the majority of subjects vaccinated with MSP2-C1 developed an antibody responses to both forms of MSP2, and that these antibodies mediated ADCI provide further support for MSP2 as a malaria vaccine candidate. However, in view of the reactogenicity of this formulation, further clinical development of MSP2-C1 will require formulation of MSP2 in an alternative adjuvant. Trial Registration: Australian New Zealand Clinical Trials Registry 12607000552482

Fetal microglial phenotype in vitro carries memory of prior in vivo exposure to inflammation

Objective. Neuroinflammation in utero may result in life-long neurological disabilities. The molecular mechanisms whereby microglia contribute to this response remain incompletely understood. Methods. Lipopolysaccharide (LPS) or saline were administered intravenously to non-anesthetized chronically instrumented near-term fetal sheep to model fetal inflammation in vivo. Microglia were then isolated from in vivo LPS and saline (naïve) exposed animals. To mimic the second hit of neuroinflammation, these microglia were then re-exposed to LPS in vitro. Cytokine responses were measured in vivo and subsequently in vitro in the primary microglia cultures derived from these animals. We sequenced the whole transcriptome of naïve and second hit microglia and profiled their genetic expression to define molecular pathways disrupted during neuroinflammation.Results. In vivo LPS exposure resulted in IL-6 increase in fetal plasma 3 h post LPS exposure. Even though not histologically apparent, microglia acquired a pro-inflammatory phenotype in vivo that was sustained and amplified in vitro upon second hit LPS exposure as measured by IL-1β response in vitro and RNAseq analyses. While NFKB and Jak-Stat inflammatory pathways were up regulated in naïve microglia, heme oxygenase 1 (HMOX1) and Fructose-1,6-bisphosphatase (FBP) genes were uniquely differentially expressed in the second hit microglia. Microglial calreticulin/LRP genes implicated in microglia-neuronal communication relevant for the neuronal development were up regulated in second hit microglia.Discussion. We identified a unique HMOX1down and FBPup phenotype of microglia exposed to the double-hit suggesting interplay of inflammatory and metabolic pathways as a memory of prior inflammatory insult. These findings suggest new therapeutic targets for early postnatal intervention to prevent brain injury