Identification of Novel CD4+ T Cell Subsets in the Target Tissue of Sjögren's Syndrome and Their Differential Regulation by the Lymphotoxin/LIGHT Signaling Axis.

Despite being one of the most common rheumatologic diseases, there is still no disease-modifying drug for primary Sjögren's syndrome (pSS). Advancing our knowledge of the target tissue has been limited by the low dimensionality of histology techniques and the small size of human salivary gland biopsies. In this study, we took advantage of a molecularly validated mouse model of pSS to characterize tissue-infiltrating CD4+ T cells and their regulation by the lymphotoxin/LIGHT signaling axis. Novel cell subsets were identified by combining highly dimensional flow and mass cytometry with transcriptomic analyses. Pharmacologic modulation of the LTßR signaling pathway was achieved by treating mice with LTßR-Ig, a therapeutic intervention currently being tested in pSS patients (Baminercept trial NCT01552681). Using these approaches, we identified two novel CD4+ T cell subsets characterized by high levels of PD1: Prdm1+ effector regulatory T cells expressing immunoregulatory factors, such as Il10, Areg, Fgl2, and Itgb8, and Il21+ effector conventional T cells expressing a pathogenic transcriptional signature. Mirroring these observations in mice, large numbers of CD4+PD1+ T cells were detected in salivary glands from Sjögren's patients but not in normal salivary glands or kidney biopsies from lupus nephritis patients. Unexpectedly, LTßR-Ig selectively halted the recruitment of PD1- naive, but not PD1+, effector T cells to the target tissue, leaving the cells with pathogenic potential unaffected. Altogether, this study revealed new cellular players in pSS pathogenesis, their transcriptional signatures, and differential dependency on the lymphotoxin/LIGHT signaling axis that help to interpret the negative results of the Baminercept trial and will guide future therapeutic interventions.

BAFF overexpression increases lymphocytic infiltration in Sjögren's target tissue, but only inefficiently promotes ectopic B-cell differentiation.

B-cell activating factor (BAFF) levels are increased in rheumatoid arthritis, lupus and primary Sjögren's syndrome (pSS). However, BAFF contribution to pathogenesis is not completely understood. In pSS, immune infiltration of the salivary and lacrimal glands leads to xerostomia and xerophtalmia. Glandular B cell hyperactivation, differentiation into germinal center (GC)-like structures and plasma cell accumulation are histopathological hallmarks that were attributed to increased BAFF. Here, we experimentally tested this hypothesis by overexpressing BAFF in a mouse model of pSS. BAFF overexpression enhanced lymphocytic infiltration and MHCII expression on B cells. Increased BAFF also induced B cell differentiation into GC B cells within the autoimmune target tissue. However, even in these conditions, GC B cells only accounted for <1% of glandular B cells, demonstrating that BAFF is not efficiently promoting ectopic GC formation in pSS and warranting further investigation of therapeutics targeting both BAFF and the related TNF-family member APRIL.

Assessment of diversity-based approaches used by American Universities to increase recruitment and retention of biomedical sciences research faculty members: A scoping review protocol.

Diversity enriches the educational experience by improving intellectual engagement, self-motivation, citizenship, cultural engagement, and academic skills like critical thinking, problem-solving, and writing for students of all races. Faculty role models from similar backgrounds are essential for students from traditionally underrepresented groups as it sends a powerful message of support, belonging, and the confidence to pursue higher education. However, in the biomedical sciences, the percentage of historically underrepresented tenure-track faculty is far lower than that of their white colleagues. For this to change, a strong strategic plan and commitment from the university are imperative. This scoping review will assess the size and scope of available peer-reviewed research literature on diversity programs that aim to increase the recruitment and retention of biomedical sciences research faculty and are implemented and evaluated at American Universities. The information provided in this scoping review will help universities identify novel, successful diversity-based approaches for recruiting and retaining biomedical science faculty that might suit their own unique academic and geographic needs and be incorporated into their diversity initiatives and policies. The review follows the Population-Concept-Context methodology for Joanna Briggs Institution Scoping Reviews. Relevant peer-reviewed studies published in English between June 1, 2012, to June 1, 2022, will be identified from the following electronic databases; MEDLINE (PubMed), Scopus (Elsevier), EMBASE (Elsevier), CINAHL (EBSCO), and ERIC (EBSCO). The search strings using the key variables "biomedical research faculty," "recruitment/retention," "diversity/ minority/ underrepresented, and "mentoring" will be conducted using Boolean logic. Two independent reviewers will conduct all title and abstract screening, followed by a full article screening and data extraction. Due to the possible heterogeneity of the studies, we hope to use either a narrative analysis and/or descriptive figures/tables to depict the results.

RAS-inhibiting biologics identify and probe druggable pockets including an SII-α3 allosteric site.

RAS mutations are the most common oncogenic drivers across human cancers, but there remains a paucity of clinically-validated pharmacological inhibitors of RAS, as druggable pockets have proven difficult to identify. Here, we identify two RAS-binding Affimer proteins, K3 and K6, that inhibit nucleotide exchange and downstream signaling pathways with distinct isoform and mutant profiles. Affimer K6 binds in the SI/SII pocket, whilst Affimer K3 is a non-covalent inhibitor of the SII region that reveals a conformer of wild-type RAS with a large, druggable SII/α3 pocket. Competitive NanoBRET between the RAS-binding Affimers and known RAS binding small-molecules demonstrates the potential to use Affimers as tools to identify pharmacophores. This work highlights the potential of using biologics with small interface surfaces to select unseen, druggable conformations in conjunction with pharmacophore identification for hard-to-drug proteins.

The periosteal requirement and temporal dynamics of BMP2-induced middle phalanx regeneration in the adult mouse.

Regeneration of mammalian limbs is restricted to amputation of the distal digit tip, the terminal phalanx (P3). The adjacent skeletal element, the middle phalanx (P2), has emerged as a model system to investigate regenerative failure and as a site to test approaches aimed at enhancing regeneration. We report that exogenous application of bone morphogenetic protein 2 (BMP2) stimulates the formation of a transient cartilaginous callus distal to the amputation plane that mediates the regeneration of the amputated P2 bone. BMP2 initiates a significant regeneration response during the periosteal-derived cartilaginous healing phase of P2 bone repair, yet fails to induce regeneration in the absence of periosteal tissue, or after boney callus formation. We provide evidence that a temporal component exists in the induced regeneration of P2 that we define as the "regeneration window." In this window, cells are transiently responsive to BMP2 after the amputation injury. Simple re-injury of the healed P2 stump acts to reinitiate endogenous bone repair, complete with periosteal chondrogenesis, thus reopening the "regeneration window" and thereby recreating a regeneration-permissive environment that is responsive to exogenous BMP2 treatment.

A simplified method to assess structurally identifiable parameters in Monod-based activated sludge models.

The first step in the estimation of parameters of models applied for data interpretation should always be an investigation of the identifiability of the model parameters. In this study the structural identifiability of the model parameters of Monod-based activated sludge models (ASM) was studied. In an illustrative example it was assumed that respirometric (dissolved oxygen or oxygen uptake rates) and titrimetric (cumulative proton production) measurements were available for the characterisation of nitrification. Two model structures, including the presence and absence of significant growth for description of long- and short-term experiments, respectively, were considered. The structural identifiability was studied via the series expansion methods. It was proven that the autotrophic yield becomes uniquely identifiable when combined respirometric and titrimetric data are assumed for the characterisation of nitrification. The most remarkable result of the study was, however, that the identifiability results could be generalised by applying a set of ASM1 matrix based generalisation rules. It appeared that the identifiable parameter combinations could be predicted directly based on the knowledge of the process model under study (in ASM1-like matrix representation), the measured variables and the biodegradable substrate considered. This generalisation reduces the time-consuming task of deriving the structurally identifiable model parameters significantly and helps the user to obtain these directly without the necessity to go too deeply into the mathematical background of structural identifiability.

Modeling aerobic carbon source degradation processes using titrimetric data and combined respirometric-titrimetric data: structural and practical identifiability.

The structural and practical identifiability of a model for description of respirometric-titrimetric data derived from aerobic batch substrate degradation experiments of a C(x)H(y)O(z) carbon source with activated sludge was evaluated. The model processes needed to describe titrimetric data included substrate uptake, CO(2) production, and NH(3) uptake for biomass growth. The structural identifiability was studied using the Taylor series method and a recently proposed generalization method. It showed that combining respirometric and titrimetric data allows structural identifiability of one extra parameter combination, the biomass yield, Y(H), compared to estimation on separate data sets, on condition that the nitrogen fraction in biomass (i(XB)) is known. However, data from short-term batch substrate degradation experiments were not sufficiently informative to allow practical identification of all structurally identifiable parameters. Combining respirometry and titrimetry resulted in improvements of parameter confidence intervals compared to estimation on separate respirometric or titrimetric data sets. However, the level of the improvement seems to be substrate dependent: parameter confidence intervals improved considerably more for dextrose than for acetate degradation models. Noteworthy is the finding that the half-saturation substrate concentrations can be different depending on whether they are estimated from respirometric or titrimetric data. Moreover, this difference appears to be dependent on the carbon source considered: for dextrose, titrimetry-based K(S) values are higher than respirometry-based values while for acetate the opposite was found. It was hypothesized that this can be explained by the different point in cell metabolism where the proton production or consumption takes place, leading to a corresponding difference in timing between pH effect and oxygen consumption. Finally, the biomass yield Y(H) and the nitrogen content of the biomass i(XB) could be estimated from combined respirometric-titrimetric data obtained with addition of a known amount of carbon source. Y(H) can also be estimated from r(O) data when the initial substrate concentration S(S)(0) is known. The values found correspond to values reported in literature, but, interestingly, also seem able to reflect the occurrence of storage processes when pulses of acetate and dextrose are added.

Modeling aerobic carbon source degradation processes using titrimetric data and combined respirometric-titrimetric data: experimental data and model structure.

Experimental data are presented that resulted from aerobic batch degradation experiments in activated sludge with simple carbon sources (acetate and dextrose) as substrates. Data collection was done using combined respirometric-titrimetric measurements. The respirometer consists of an open aerated vessel and a closed non-aerated respiration chamber for monitoring the oxygen uptake rate related to substrate degradation. The respirometer is combined with a titrimetric unit that keeps the pH of the activated sludge sample at a constant value by addition of acid and/or base. The experimental data clearly showed that the activated sludge bacteria react with consumption or production of protons during aerobic degradation of the two carbon sources under study. Thus, the cumulative amount of added acid and/or base could serve as a complementary information source on the degradation processes. For acetate, protons were consumed during aerobic degradation, whereas for dextrose protons were produced. For both carbon sources, a linear relationship was found between the amount of carbon source added and the amount of protons consumed (in case of acetate: 0.38 meq/mmol) or produced (in case of dextrose: 1.33 meq/mmol) during substrate degradation. A model taking into account substrate uptake, CO(2) production, and NH(3) uptake for biomass growth is proposed to describe the aerobic degradation of a C(x)H(y)O(z)-type carbon source. Theoretical evaluation of this model for reference parameters showed that the proton effect due to aerobic substrate degradation is a function of the pH of the liquid phase. The proposed model could describe the experimental observations with both carbon sources.