Seeds of Success: Empowering Latina STEM Girl Ambassadors Through Role Models, Leadership, and STEM-Related Experiences.

Gender stereotypes and lack of access to relatable role models, mentors, and STEM opportunities have been suggested to deter middle school girls and students from underrepresented backgrounds away from STEM. Seeds of Success, an out-of-school program, is designed to inspire girls to consider STEM careers by countering gender stereotypes through relatable role models, promoting STEM confidence through STEM workshops and hands-on activities, and encouraging alignment between cultural and STEM identities through community-based STEM projects that develop leadership skills. Since 2015, the program has impacted 453 students who in turn have reached more than 42,777 people in Puerto Rico through their STEM Ambassadors projects. A robust mix-method evaluation of the 2020 and 2021 cohorts demonstrates significant improvements in participants' STEM attitudes and science identity, as well as in their self-perception as a leader, confidence in their ability to succeed in science, knowledge about STEM careers and opportunities, and access to STEM role models. Moreover, 95% of participants intend to continue participating in STEM activities after the program and overall scores for the entire survey were significantly higher after the program than before. We discuss lessons learned for other programs seeking to empower girls from historically underserved backgrounds in STEM.

Iron Chelator Transmetalative Approach to Inhibit Human Ribonucleotide Reductase.

Efforts directed at curtailing the bioavailability of intracellular iron could lead to the development of broad-spectrum anticancer drugs given the metal's role in cancer proliferation and metastasis. Human ribonucleotide reductase (RNR), the key enzyme responsible for synthesizing the building blocks of DNA replication and repair, depends on Fe binding at its R2 subunit to activate the catalytic R1 subunit. This work explores an intracellular iron chelator transmetalative approach to inhibit RNR using the titanium(IV) chemical transferrin mimetic (cTfm) compounds Ti(HBED) and Ti(Deferasirox)2. Whole-cell EPR studies reveal that the compounds can effectively attenuate RNR activity though seemingly causing different changes to the labile iron pool that may account for differences in their potency against cells. Studies of Ti(IV) interactions with the adenosine nucleotide family at pH 7.4 reveal strong metal binding and extensive phosphate hydrolysis, which suggest the capacity of the metal to disturb the nucleotide substrate pool of the RNR enzyme. By decreasing intracellular Fe bioavailability and altering the nucleotide substrate pool, the Ti cTfm compounds could inhibit the activity of the R1 and R2 subunits of RNR. The compounds arrest the cell cycle in the S phase, indicating suppressed DNA replication, and induce apoptotic cell death. Cotreatment cell viability studies with cisplatin and Ti(Deferasirox)2 reveal a promising synergism between the compounds that is likely owed to their distinct but complementary effect on DNA replication.

An exploratory study comparing students' science identity perceptions derived from a hands-on research and nonresearch-based summer learning experience.

Although multiple efforts have been initiated to increase students' science proficiency scores, most of the schools in the United States do not reach the expected student academic performance. This study addresses the impact of a one-week summer scientific learning experience on students that worked with experimental procedures and students that did not. We describe and evaluate these two different interventions to explore what components influence high school students' perception of their scientific competence, performance, and recognition, using science identity as an analytical lens. Science identity score was increased at the end of both interventions. Interestingly, science identity change index was higher for the group that did not work with experimental procedures. Although this group did not perform any hands-on experiments, they report, through reflexive diaries and interviews that working with CRISPR-Cas9 models, being in a research laboratory, and seeing the instrumentation made them feel like scientists. Regarding science competence, both groups report exponential learning gains, although the group that performed the experiments reports more difficulties. Both groups report that mentorship was key in their competence and performance development. These findings suggest that our one-week scientific learning programs influence participants' perception of scientific competence and performance and create an opportunity to develop further studies on short scientific learning experiences using models and active learning activities.

The Characterization of Monoclonal Antibodies to Mouse TLT-1 Suggests That TLT-1 Plays a Role in Wound Healing.

Platelets play a vital role in hemostasis and inflammation. The membrane receptor TREM-like transcript-1 (TLT-1) is involved in platelet aggregation, bleeding, and inflammation, and it is localized in the α-granules of platelets. Upon platelet activation, TLT-1 is released from α-granules both in its transmembrane form and as a soluble fragment (sTLT-1). Higher levels of sTLT-1 have been detected in the plasma of patients with acute inflammation or sepsis, suggesting an important role for TLT-1 during inflammation. However, the roles of TLT-1 in hemostasis and inflammation are not well understood. We are developing the mouse model of TLT-1 to mechanistically test clinical associations of TLT-1 in health and disease. To facilitate our studies, monoclonal murine TLT-1 (mTLT-1) antibodies were produced by the immunization of a rabbit using the negatively charged region of the mTLT-1 extracellular domain 122PPVPGPREGEEAEDEK139. In the present study, we demonstrate that two selected clones, 4.6 and 4.8, are suitable for the detection of mTLT-1 by western blot, immunoprecipitation, immunofluorescent staining, flow cytometry and inhibit platelet aggregation in aggregometry assays. In addition, we found that the topical administration of clone 4.8 delayed the wound healing process in an experimental burn model. These results suggest that TLT-1 plays an important role in wound healing and because both clones specifically detect mTLT-1, they are suitable to further develop TLT-1 based models of inflammation and hemostasis in vivo.

The Rise of CRISPR/Cas for Genome Editing in Stem Cells.

Genetic manipulation is a powerful tool to establish the causal relationship between a genetic lesion and a particular pathological phenotype. The rise of CRISPR/Cas9 genome-engineering tools overcame the traditional technical bottleneck for routine site-specific genetic manipulation in cells. To create the perfect in vitro cell model, there is significant interest from the stem cell research community to adopt this fast evolving technology. This review addresses this need directly by providing both the up-to-date biochemical rationale of CRISPR-mediated genome engineering and detailed practical guidelines for the design and execution of CRISPR experiments in cell models. Ultimately, this review will serve as a timely and comprehensive guide for this fast developing technology.