Covariational reasoning and item context affect language in undergraduate mass balance written explanations.

Mass balance (MB) reasoning offers a rich topic for examination of students' scientific thinking and skills, as it requires students to account for multiple inputs and outputs within a system and apply covariational reasoning. Using previously validated constructed response prompts for MB, we examined 1,920 student-constructed responses (CRs) aligned to an emerging learning progression to determine how student language changes from low (1) to high (4) covariational reasoning levels. As students' abilities and thinking change with Context, we used the same general prompt in six physiological contexts. We asked how Level and Context affect student language and what language is conserved across Contexts at higher reasoning Levels. Using diversity methods, we found student language becomes more similar as covariational reasoning level increases. Using text analysis, we found context-dependent words at each Level; however, the type of context words changed. Specifically, at Level 1, students used context words that are tangential to MB reasoning, while Level 4 responses used words that specify inputs and outputs for the given Item Context. Further, at Level 4, students shared 30% of language across the six contexts and leveraged context-independent words including rate, equal, and some form of slower/lower/smaller. Together, these data demonstrate that Context affects undergraduate MB language at all covariational reasoning levels, but that the language becomes more specific and similar as Level increases. These findings encourage instructors to foster context-independent, comparative, and summative language during instruction to functionally build MB and covariational reasoning skills across contexts.NEW & NOTEWORTHY This article builds on the work of Scott et al. (Scott EE, Cerchiara J, McFarland JL, Wenderoth MP, Doherty JH. J Res Sci Teach 1: 37, 2023) and Shiroda et al. (Shiroda M, Fleming MP, Haudek KC. Front Educ 8: 989836, 2023) to quantitatively examine student language in written explanations of mass balance across six contexts using constructed response assessments. These results present an evaluation of student mass balance language and provide researchers and practitioners with tools to assist students in constructing scientific mass balance reasoning explanations.

Lactobacillus strains vary in their ability to interact with human endometrial stromal cells.

The placental membranes that surround the fetus during pregnancy were suggested to contain a low abundance microbiota. Specifically, abundance of Lactobacillus, a probiotic and dominant member of the microbiome of the lower reproductive tract, has been shown to correlate with healthy, term pregnancies. We therefore sought to assess the interactions between four different Lactobacillus strains with immortalized decidualized endometrial cells (dT-HESCs), which were used as a model to represent the outermost layer of the placental membranes. Notably, we demonstrated that all four Lactobacillus strains could associate with dT-HESCs in vitro. L. crispatus was significantly more successful (p < 0.00005), with 10.6% of bacteria attaching to the host cells compared to an average of 0.8% for the remaining three strains. The four strains also varied in their ability to form biofilms. Dependent on media type, L. reuteri 6475 formed the strongest biofilms in vitro. To examine the impact on immune responses, levels of total and phosphorylated protein p38, a member of the Mitogen Activated Protein Kinase (MAPK) pathway, were examined following Lactobacillus association with dT-HESCs. Total levels of p38 were reduced to an average of 44% that of the cells without Lactobacillus (p < 0.05). While a trend towards a reduction in phosphorylated p38 was observed, this difference was not significant (p > 0.05). In addition, association with Lactobacillus did not result in increased host cell death. Collectively, these data suggest that varying types of Lactobacillus can attach to the outermost cells of the placental membranes and that these interactions do not contribute to inflammatory responses or host cell death. To our knowledge this is the first in vitro study to support the ability of Lactobacillus to interact with placental cells, which is important when considering its use as a potential probiotic within the reproductive tract.

The impact of Lactobacillus on group B streptococcal interactions with cells of the extraplacental membranes.

Group B Streptococcus (GBS) causes adverse pregnancy outcomes and neonatal disease. The recommended preventative measure is intrapartum antibiotic prophylaxis, which can prevent early onset neonatal disease but not chorioamnionitis, preterm labor, stillbirth, or late-onset disease. Novel prevention methods are therefore needed. Use of probiotics including Lactobacillus spp., has been suggested given that they are dominant members of the lower reproductive tract microbiome. Although Lactobacillus was shown to reduce recto-vaginal colonization of GBS, no studies have examined how Lactobacillus impacts GBS in the extraplacental membranes. Since Lactobacillus has been detected in the placental membranes, we sought to characterize GBS-Lactobacillus interactions in vitro using a colonizing and invasive GBS strain. While live Lactobacillus did not affect growth or biofilms in GBS, co-culture with L. gasseri led to a 224-fold increase in GBS association with decidualized human endometrial stromal cells for both GBS strains (p < 0.005). Increased association did not result in increased invasion (p > 0.05) or host cell death, though some GBS and Lactobacillus combinations contributed to a significant reduction in host cell death (p < 0.05). Since Lactobacillus secretes many inhibitory compounds, the effect of Lactobacillus supernatants on GBS was also examined. The supernatants inhibited GBS growth, biofilm formation and invasion of host cells, though strain dependent effects were observed. Notably, supernatant from L. reuteri 6475 broadly inhibited growth in 36 distinct GBS strains and inhibited GBS growth to an average of 46.6% of each GBS strain alone. Together, these data show that specific Lactobacillus strains and their secreted products have varying effects on GBS interactions with cells of the extraplacental membranes that could impact pathogenesis. Understanding these interactions could help guide new treatment options aimed at reducing GBS-associated maternal complications and disease.

RpoS impacts the lag phase of Salmonella enterica during osmotic stress.

Salmonella enterica can survive harsh environmental conditions, including hyperosmotic stress. It is well established that the alternative sigma factor, σ(s) (RpoS), is required for maximal survival of enteric pathogens, including S. enterica. Although RpoS levels are greatest during stationary phase or stress conditions, RpoS can be found in S. enterica during growth. However, its activity during growth is poorly characterized. In this study, the impact of RpoS levels on the growth of S. enterica in LB supplemented with 6% NaCl (LB-NaCl) was examined. Cells in stationary phase prior to inoculation into LB-NaCl had a shorter lag phase than did exponential-phase cells. In addition, the deletion of rpoS from S. enterica Typhimurium M-09 (M-09 ΔrpoS) increased the length of lag phase in LB-NaCl relative to the parental strain. Complementation of M-09 ΔrpoS in trans by an inducible plasmid encoding rpoS reduced the length of lag phase. The length of lag phase in both the rpoS mutant and complemented strain was independent of their growth phase prior to inoculation of LB-NaCl. The results from this study demonstrate that the level of RpoS influences the length of lag phase and the growth of S. enterica in hyperosmotic growth conditions.