Mustafa G Mujtaba

Getting real-time feedback on pedagogical methods used in the classroom can be a challenge due to a variety of reasons, including the classroom format, instructor lecturing styles, as well as weariness of students to engage instructors. However, a structured semester-long student-faculty partnership can serve as a powerful tool to enhance teaching effectiveness and student engagement. We describe here our version of a student-faculty partnership program (SFPP) model in an upper-level undergraduate Immunology course where a student from a non-science major was embedded in the course to observe classroom dynamics between students and instructor and provide formative feedback to the instructor throughout the semester. This collaboration led to several pedagogical adjustments suggested by the student-partner, including increased use of flipped classroom sessions, integration of lab experiments within lecture sessions, and the use of interactive tools, such as clickers for concept reinforcement and game apps for exam reviews. As a result, the course section implementing the SFPP model had high engagement in lectures and labs, increased performance on lab assignment submitted, and better satisfaction with the revised course format. The end of the semester student surveys and comments indicated student enthusiasm and comfort with the course content as a result of the revised instructional strategies. Thus, the SFPP model offers an adaptable approach for instructors seeking to create a more responsive, real-time, and student-centered learning environment in immunology and biology education.

Abstract Description
 Brevetoxins are marine algal biotoxins present in warm coastal regions during red tide blooms and bind to and modulate voltage-gated sodium ion channel (VGSC) function on neuronal cells to confer their deleterious activity. In this study, we evaluate the effect of PbTx-2, the most prevalent type of brevetoxins produced during red tide blooms, on macrophage (RAW 264.7) inflammatory responses using ELISA, nitric oxide assay, as well as toxicity assays, and determine if VGSCs play a part in brevetoxin’s mechanism via patch-clamp analysis. Brevetoxin (< 2000 ng/mL) was not toxic to the macrophage cell line when incubated for as long as 24 hours. In addition, brevetoxin did not inhibit or enhanced the lipopolysaccharide (LPS) induction of nitric oxide as well as TNFa production by the RAW 264.7 macrophages. Brevetoxin did, however, stimulate TNFa production in the macrophages in a dose-dependent manner when incubated alone with the cells. Furthermore, brevetoxin inhibited LPS stimulation of IL-1 and IL-6. VGSCs are not expressed on the plasma membrane of RAW 264.7 macrophage cells in the naïve or LPS-activated state as no sodium ion current flow could be measured upon test pulse depolarizations in patch-clamp experiments. Thus, brevetoxin modulates inflammatory responses in RAW 264.7 mouse macrophages differentially, and these effects are not due to binding of brevetoxin to VGSCs on the surface of RAW 264.7 macrophage cells since sodium ion current could not be detected.
Funding Sources
Supported by the Holmes Development Funds Grant managed by Whitaker Center for STEM Education at Florida Gulf Coast University.
Topic Categories
Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)

Macrophages are leukocytes that are an integral part of the innate immune system. Recently, we have shown that the red tide toxin, brevetoxin, and the local anesthetic, bupivacaine, modulate the release of inflammatory cytokines from RAW 264.7 macrophage cells. Both bupivacaine and brevetoxin bind to voltage-gated sodium ion channels (VGSCs), found on the plasma membrane. In this study, we determine if the mechanism of action of brevetoxin and bupivacaine on macrophage cells is through the modulation of VGSCs by conducting whole-cell patch-clamp electrophysiology experiments. Activation and inactivation patch-clamp protocols were conducted to measure gating properties of VGSCs on RAW 264.7 macrophages in the presence and absence of lipopolysaccharide, a macrophage activator, as well as control neuronal pituitary GH3 cells that are known to express native VGSCs. Our study reveals that GH3 cell VGSCs are affected by both bupivacaine and brevetoxin. Bupivacaine served as a sodium channel antagonist as sodium ion currents were blocked in GH3 cells. Unlike bupivacaine, brevetoxin activated the VGSCs in GH3 cells. On the other hand, it was shown that macrophage cells do not express any VGSCs on their plasma membrane as no sodium ion currents were detected upon depolarizing test pulses. The data in this study illustrates that brevetoxin and bupivacaine effects on inflammatory responses from macrophage cells are not due to modulation of plasma membrane bound VGSCs.
 
Macrophages are leukocytes that are an integral part of the innate immune system. Recently, we have shown that the red tide toxin, brevetoxin, and the local anesthetic, bupivacaine, modulate the release of inflammatory cytokines from RAW 264.7 macrophage cells. Both bupivacaine and brevetoxin bind to voltage-gated sodium ion channels (VGSCs), found on the plasma membrane. In this study, we determine if the mechanism of action of brevetoxin and bupivacaine on macrophage cells is through the modulation of VGSCs by conducting whole-cell patch-clamp electrophysiology experiments. Activation and inactivation patch-clamp protocols were conducted to measure gating properties of VGSCs on RAW 264.7 macrophages in the presence and absence of lipopolysaccharide, a macrophage activator, as well as control neuronal pituitary GH3 cells that are known to express native VGSCs. Our study reveals that GH3 cell VGSCs are affected by both bupivacaine and brevetoxin. Bupivacaine served as a sodium channel antagonist as sodium ion currents were blocked in GH3 cells. Unlike bupivacaine, brevetoxin activated the VGSCs in GH3 cells. On the other hand, it was shown that macrophage cells do not express any VGSCs on their plasma membrane as no sodium ion currents were detected upon depolarizing test pulses. The data in this study illustrates that brevetoxin and bupivacaine effects on inflammatory responses from macrophage cells are not due to modulation of plasma membrane bound VGSCs.
Funding Sources
This study was supported by the Holmes Development Fund Grant from the Whitaker Center at FGCU
Topic Categories
Innate Immune Responses and Host Defense: Molecular Mechanisms (INM)

The impact of acculturation on leadership style is an important topic
for the global community. A quantitative, non-experimental,
descriptive-correlational design using primary retrospective data was
employed in this study. The objective of the study was to see if
gender, education, income, primary language spoken at home, level
of Afghan acculturation, and level of American acculturation predict
task- and relationship-oriented leadership styles. Respondents were
Afghan American adults currently working in the U.S. Health System
as registered nurses with one or both parents originally from
Afghanistan, N = 171 participants. The results showed that levels of
task-oriented leadership correspond to the span of control (number of
subordinates overseen) directly. The more subordinates, the greater
the tendencies toward task-oriented leadership. The fewer
subordinates, the lower the tendencies toward task-oriented
leadership. None of the other variables contributed significantly to
predicting task-oriented leadership. Due to the constant changing
nursing field, diverse workforce, and high demand for healthcare
administrators, nursing curriculum should emphasize the importance of leadership skills. Implications, limitations, and directions for future research are discussed.

The Ouchterlony double immunodiffusion assay is a serological technique used in the detection of antibodies and antigens for diagnostic purposes and also used in immunology laboratory courses as a common teaching assay where students observe the geometrical precipitation line patterns that form in the agarose, elucidating degrees of homology between antigens. In this classical technique, students must wait several hours to days to obtain results when protein antigens and antibodies are used.
ABSTRACT The Ouchterlony double immunodiffusion assay is a serological technique used in the detection of antibodies and antigens for diagnostic purposes and also used in immunology laboratory courses as a common teaching assay where students observe the geometrical precipitation line patterns that form in the agarose, elucidating degrees of homology between antigens. In this classical technique, students must wait several hours to days to obtain results when protein antigens and antibodies are used. Furthermore, these proteins degrade over time if not frozen or stored in the refrigerator and are the most expensive consumables of the laboratory exercise. In this study, inexpensive and commonly used inorganic ionic salt solutions that are stable and can be stored at room temperature for several years were used to mimic antigens and antibodies. The precipitation lines started to form in the agarose plates after 15 min and fully developed within an hour, showing different geometrical precipitation patterns and spur formations that could be identified by students as full identity, partial identity, and nonidentity between the simulated (inorganic) antigens. Students conducting this exercise in a combined lecture and laboratory immunology course were able to finish the exercise as well as record and discuss results within class time, and tvhey showed increased interest in the laboratory exercise and had a better understanding of antibody-antigen reactions. Thus, this simulated laboratory experiment is an inexpensive, safe, and fast exercise that allows students to observe precipitations reactions of the Ouchterlony assay within the class session time.

The staphylococcal enterotoxins (SEs) are classified as superantigens due to their potent stimulation of the immune system resulting in T cell activation and prodigious cytokine production and toxicity. This study examined the ability of superantigens to induce prophylactic antiviral activity and and evaluated potential superantigen mimetic peptides. Prophylactic treatment of mice with intraperitoneal injections of SE superantigens SEA and SEB (both at 20  g/day for 3 days) prevented encephalomyocarditis virus (EMCV)-induced lethality in 100% and 80% of mice, respectively, as compared with control saline-treated groups in which EMCV was lethal to all mice. Furthermore, SEA (2  g/mL) and SEB (1  g/mL) induced antiviral activity in mouse splenocytes to produce an antiviral factor since their supernatant prevented EMCV lysis of L929 cells in tissue culture. It was found that superantigens do not directly prevent EMCV infection, but rather indirectly through inducing interferon gamma (IFN ) production in cells as the antiviral factor. Evaluation of various superantigen mimetic peptides showed that one peptide (SEA3) had superantigen-like activity by inducing IFN production in cells but without the cellular proliferation, as associated with superantigens. However, the induction of IFN activation by the SEA3 peptide was not as pronounced, and took a much higher peptide concentration, when compared with the parent superantigen. If the negative side effects of superantigens can be eliminated, their beneficial properties can be harnessed for prophylactic treatment of viral infections and other pathologies requiring a robust immune response.

The impact of acculturation on leadership style is an important topic for the global community, especially during stressful years of the Covid-19 pandemic. A quantitative, non-experimental, descriptive-correlational design using primary retrospective data was employed in this study. We used established instruments such as the Acculturation Rating Scale Questionnaire and the Leadership Style Questionnaire (LSQ) with 172 working adult respondents in the U.S. Health System as registered nurses. The results showed that Afghan male and female nurses tended to identify with the Afghan culture or the American culture, not both. Afghan male nurses’ relationship-oriented leadership was related to both Afghan and (more weakly) American acculturation (the more acculturation, the more relationship orientation).

All undergraduates majoring in a physical or natural science in the College of Arts and Sciences at Florida Gulf Coast University (FGCU) are required to take an interdisciplinary-science methods course entitled Scientific Process. This course is designed to help professionalize students by introducing them to the history, practice, philosophy, and ethics associated with being a working scientist (Meers, Demers and Savarese 2003). Most students take the course early in their junior year as the first class in a sequence of research courses that culminates in conducting and presenting independent research during their senior year.

Suppressor of cytokine signaling (SOCS)-1 protein modulates signaling by IFN-gamma by binding to the autophosphorylation site of JAK2 and by targeting bound JAK2 to the proteosome for degradation. We have developed a small tyrosine kinase inhibitor peptide (Tkip) that is a SOCS-1 mimetic. Tkip is compared in this study with the kinase inhibitory region (KIR) of SOCS-1 for JAK2 recognition, inhibition of kinase activity, and regulation of IFN-gamma-induced biological activity. Tkip and a peptide corresponding to the KIR of SOCS-1, ((53))DTHFRTFRSHSDYRRI((68)) (SOCS1-KIR), both bound similarly to the autophosphorylation site of JAK2, JAK2(1001-1013). The peptides also bound to JAK2 peptide phosphorylated at Tyr(1007), pJAK2(1001-1013). Dose-response competitions suggest that Tkip and SOCS1-KIR similarly recognize the autophosphorylation site of JAK2, but probably not precisely the same way. Although Tkip inhibited JAK2 autophosphorylation as well as IFN-gamma-induced STAT1-alpha phosphorylation, SOCS1-KIR, like SOCS-1, did not inhibit JAK2 autophosphorylation but inhibited STAT1-alpha activation. Both Tkip and SOCS1-KIR inhibited IFN-gamma activation of Raw 264.7 murine macrophages and inhibited Ag-specific splenocyte proliferation. The fact that SOCS1-KIR binds to pJAK2(1001-1013) suggests that the JAK2 peptide could function as an antagonist of SOCS-1. Thus, pJAK2(1001-1013) enhanced suboptimal IFN-gamma activity, blocked SOCS-1-induced inhibition of STAT3 phosphorylation in IL-6-treated cells, enhanced IFN-gamma activation site promoter activity, and enhanced Ag-specific proliferation. Furthermore, SOCS-1 competed with SOCS1-KIR for pJAK2(1001-1013). Thus, the KIR region of SOCS-1 binds directly to the autophosphorylation site of JAK2 and a peptide corresponding to this site can function as an antagonist of SOCS-1.

Suppressor of cytokine signaling (SOCS)-1 protein modulates signaling by IFN-γ by binding to the autophosphorylation site of JAK2 and by targeting bound JAK2 to the proteosome for degradation. We have developed a small tyrosine kinase inhibitor peptide (Tkip) that is a SOCS-1 mimetic. Tkip is compared in this study with the kinase inhibitory region (KIR) of SOCS-1 for JAK2 recognition, inhibition of kinase activity, and regulation of IFN-γ-induced biological activity. Tkip and a peptide corresponding to the KIR of SOCS-1, (53)DTHFRTFRSHSDYRRI(68) (SOCS1-KIR), both bound similarly to the autophosphorylation site of JAK2, JAK2(1001–1013). The peptides also bound to JAK2 peptide phosphorylated at Tyr1007, pJAK2(1001–1013). Dose-response competitions suggest that Tkip and SOCS1-KIR similarly recognize the autophosphorylation site of JAK2, but probably not precisely the same way. Although Tkip inhibited JAK2 autophosphorylation as well as IFN-γ-induced STAT1-α phosphorylation, SOCS1-KIR, like SOCS-1, did not inhibit JAK2 autophosphorylation but inhibited STAT1-α activation. Both Tkip and SOCS1-KIR inhibited IFN-γ activation of Raw 264.7 murine macrophages and inhibited Ag-specific splenocyte proliferation. The fact that SOCS1-KIR binds to pJAK2(1001–1013) suggests that the JAK2 peptide could function as an antagonist of SOCS-1. Thus, pJAK2(1001–1013) enhanced suboptimal IFN-γ activity, blocked SOCS-1-induced inhibition of STAT3 phosphorylation in IL-6-treated cells, enhanced IFN-γ activation site promoter activity, and enhanced Ag-specific proliferation. Furthermore, SOCS-1 competed with SOCS1-KIR for pJAK2(1001–1013). Thus, the KIR region of SOCS-1 binds directly to the autophosphorylation site of JAK2 and a peptide corresponding to this site can function as an antagonist of SOCS-1.