Communicating Branches of the Facial Nerve: Descriptions and Clinical Considerations.

BACKGROUND: Major branching patterns of the facial nerve have been extensively studied because damage to branches of the nerve is associated with complications ranging from weakness to paralysis. However, communicating branches of the facial nerve have received far less attention despite being hypothesized as a means of motor recovery following facial nerve injury. OBJECTIVES: The aim of this study was to characterize the frequency of communicating branches of the facial nerve to provide clarity on their anatomy and clinical correlations. METHODS: Bilateral facial dissections were completed on cadaveric donors (n = 20) to characterize the frequency and location of communicating branches across terminal branches of the facial nerve. Statistical analyses were employed to analyze differences between the location of communications by side and whether the communicating branches were more likely to occur on the left or right side (P < 0.05). RESULTS: Communicating branches were identified among all terminal branches of the facial nerve and their frequencies reported. The highest frequencies of communicating branches were identified between the buccal-to-marginal mandibular and zygomatic-to-buccal branches, at 67.5% (27 comm/40 hemifaces). The second highest frequency was identified between the temporal-to-zygomatic branches in 62.5% (25/40) of donors. The marginal mandibular-to-cervical branches had communicating branches at a frequency of 55% (22/40). Location or sidedness of communicating branches did not significantly differ. CONCLUSIONS: Our characterization more accurately defines generalizable areas in which communicating branches are located. These locations of branches, described in relation to nearby landmarks, are fundamental for clinical and surgical settings to improve procedural awareness.

Osteopathic Manipulative Treatment as an Intervention to Reduce Stress, Anxiety, and Depression in First Responders: A Pilot Study.

In this pilot study, we examined the efficacy of Osteopathic Manipulative Treatment (OMT) for improving symptoms of stress, anxiety, and depression (SAD) to determine a correlation between overall improvement in health and quality of life for first responders. Participants received weekly OMT or sham OMT targeting autonomic imbalance. Indicators of SAD were examined pre- and post-study. Overall, this pilot study suggests improvement in both the social-psychological (mental) self-assessments, and alterations in SAD-associated biomarkers from OMT.

Destination integration: linking physiology, histology, and embryology content in foundational sciences.

Anatomy and physiology are tightly linked disciplines that complement each other, however, in medical education delivery of this content is often siloed and divided. To address this, we created combined anatomy and physiology content for the female reproductive system, and team-taught designated histology and embryology topics integrated with the physiology content. Collectively, this created a more holistic incorporation of topics for student learning. Here we describe the format and approach for this teaching innovation.

Student perceptions of remote versus on-campus gross anatomy laboratories during COVID-19.

At Kansas City University, anatomy laboratories were delivered via remote (REM) or on-campus (OC) formats due to COVID-19, creating an opportunity to evaluate student perceptions of differences in laboratory delivery format. A six-item survey of Likert scale and open-ended questions explored the utility of anatomy software, prelab instruction handouts, and prosection reviews. Likert scale validity was analyzed using Cronbach's α; responses were compared among REM and OC formats using Chi-square. Descriptive codes were applied to summarize responses, which were grouped and converted into percentages. Statistically significant differences in REM versus OC formats were determined for the helpfulness of the prelab handouts (χ2 , 28.00; df, 4; p < 0.001) and effectiveness of cadavers in learning anatomy (χ2 , 20.58; df, 4; p < 0.0004). Trends in responses noted disagreement in the effectiveness of anatomy software (REM, 69.8%; OC, 51.08%), but agreement with the helpfulness of prosection reviews (REM, 85.9%; OC, 61.6%) (Cronbach α: REM, 0.648; OC, 0.646). Themes from narrative REM comments (n = 496) noted anatomy software was difficult to use (33.1%) and had issues with orientation (15.5%), as well as a student preference for OC laboratories (12.5%). The OC format responses (n = 456) noted poor software design (47.9%), unnecessary for studying (35.4%), and preference for in-person laboratories (7.4%). Qualitative analysis of narrative comments detailed other resources used, including Complete Anatomy™ and YouTube™. Trends highlighted the prelab handouts and prosection reviews for learning, the ineffectiveness of anatomy software, and a preference for OC laboratories. We highlight student perspectives of REM versus OC laboratory formats in response to COVID-19.

COVID-19 vaccine hesitancy: Considerations for reluctance and improving vaccine uptake.

The emergence of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; COVID-19) pandemic during the fall of 2019 led to the rapid development of vaccines aimed at curbing viral infection, spread, and its potential eradication. A recent trend is an overall increase in vaccine hesitancy, leading to the World Health Organization citing this as a problem which needs to be addressed. With the development and approval of vaccines for COVID-19, this trend has quickened, leading to potential negative ramifications in the ability controlling COVID-19 spread. Here we describe reported examples in overall vaccine hesitancy prior to the emergence of COVID-19, as well as summarizing recent reports on vaccine hesitancy related to COVID-19 vaccines. Gaining a better understanding of the reasons individuals have, as well as potential methods for decreasing hesitancy in the future, will hopefully lead to a greater percentage of vaccinated individuals and aid in ending the current pandemic.

Sterilization and sanitizing of 3D-printed personal protective equipment using polypropylene and a Single Wall design.

BACKGROUND: The emergence of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic during the fall of 2019 and into the spring of 2020 has led to an increased demand of disposable N95 respirators and other types of personal protective equipment (PPE) as a way to prevent virus spread and help ensure the safety of healthcare workers. The sudden demand led to rapid modification, development, and dissemination of 3D printed PPE. The goal of this study was to determine the inherent sterility and re-sterilizing ability of 3D printed PPE in order to provide sterile equipment to the healthcare field and the general public. METHODS: Samples of polylactic acid (PLA), thermoplastic polyurethane (TPU) (infill-based designs) and polypropylene (single-wall hollow design) were 3D printed. Samples were inoculated with E. coli for 24 h and then sanitized using various chemical solutions or heat-based methods. The samples were then incubated for 24- or 72-h in sterile LB medium at 37°C, and bacterial growth was measured by optical density at 600nm. Statistical analysis was conducted using GraphPad Prism v8.2.1. RESULTS: Significant bacterial growth was observed in all PLA and TPU based samples following re-sterilization, regardless of the methods used when compared to controls (p < 0.05). The single-walled hollow polypropylene design was not only sterile following printing, but was also able to undergo re-sanitization following bacterial inoculation, with no significant bacterial growth (p > 0.05) observed regardless of sanitization method used. CONCLUSION: The cost effectiveness, ease of sanitization, and reusability of 3D printed PPE, using our novel single-walled polypropylene design can help meet increased demands of PPE for healthcare workers and the general public that are needed to help decrease the viral transmission of the coronavirus disease of 2019 (COVID-19) pandemic. 3D printing also has the potential to lead to the creation and production of other sterile material items for the healthcare industry in the future. The ability to re-sterilize 3D printed PPE, as our design shows, would also contribute less to the increase in biomedical waste (BMW) being experienced by COVID-19.

Temporally and spatially controlled expression of transgenes in embryonic and adult tissues.

Using ES cell-mediated transgenesis, we generated a novel mouse strain that permits a temporally and spatially controlled expression of responder genes in embryonic and multiple adult tissues. The transgene was constructed in a way that a CMV enhancer linked to the chicken beta-actin promoter (CAG) drives the expression of the tetracycline-controlled transactivator (tTA) in particular tissues upon Cre-mediated excision of a floxed betageo marker located between the promoter and the tTA. Based on the enzymatic activity of lacZ, the CAG-betageo-tTA construct exhibits a widespread expression and appears to be very strong in the brain, heart, muscle, pancreas, and skin. Like the embryonic stem cell line that was used to generate this strain, the CAG-betageo-tTA transgene is already highly active in preimplantation embryos. Using in vivo bioluminescence imaging on MMTV-Cre, CAG-betageo-tTA, TetO-Luciferase triple transgenic mice and their controls, we demonstrated that the expression of the tTA, which is strictly dependent on the presence of Cre recombinase, induces the activation of the reporter transgene in the absence of any ligands. The tTA-mediated transactivation can be completely ablated through administration of doxycycline, and its subsequent withdrawal lifts the transcriptional block. Based on these characteristics, this novel strain may be useful in experiments that require a sustained expression of transgenes in particular cell types over a prolonged period followed by a rapid downregulation, for example in studies that examine the therapeutic value of cancer-initiating oncogenes during disease progression.

Associations between Pregnane X Receptor and Breast Cancer Growth and Progression.

Pregnane X receptor (PXR, NR1I2) is a member of the ligand-activated nuclear receptor superfamily. This receptor is promiscuous in its activation profile and is responsive to a broad array of both endobiotic and xenobiotic ligands. PXR is involved in pivotal cellular detoxification processes to include the regulation of genes that encode key drug-metabolizing cytochrome-P450 enzymes, oxidative stress response, as well as enzymes that drive steroid and bile acid metabolism. While PXR clearly has important regulatory roles in the liver and gastrointestinal tract, this nuclear receptor also has biological functions in breast tissue. In this review, we highlight current knowledge of PXR's role in mammary tumor carcinogenesis. The elevated level of PXR expression in cancerous breast tissue suggests a likely interface between aberrant cell division and xeno-protection in cancer cells. Moreover, PXR itself exerts positive effect on the cell cycle, thereby predisposing tumor cells to unchecked proliferation. Activation of PXR also plays a key role in regulating apoptosis, as well as in acquired resistance to chemotherapeutic agents. The repressive role of PXR in regulating inflammatory mediators along with the existence of genetic polymorphisms within the sequence of the PXR gene may predispose individuals to developing breast cancer. Further investigations into the role that PXR plays in driving tumorigenesis are needed.

Longitudinal analysis of mammogenesis using a novel tetracycline-inducible mouse model and in vivo imaging.

We generated a novel mouse model, which expresses the tetracycline-inducible transactivator under the regulation of the endogenous whey acidic protein gene. Using a tet-responsive luciferase reporter transgene, we demonstrated that the Wap-rtTA knockin allele allows a tightly controlled temporal and spatial expression of transgenes in the mammary gland in a ligand-inducible manner. The longitudinal analysis of individual females throughout their reproductive cycles using in vivo bioluminescence imaging confirmed that the expression of the Wap-rtTA knockin allele is highly upregulated during lactation. However, the extent of the transcriptional activation of the targeted Wap locus is dependent on the suckling stimulus and milk retrieval. In addition, we used WAP-rtTA/TetO-H2B-GFP double-transgenic females to monitor the presence of GFP-labeled parity-induced mammary epithelial cells (PI-MECs) during the postlactational involution period. The study shows that, unlike their progeny in mammary epithelial transplants as reported previously, PI-MECs themselves may not belong to the long-term label-retaining epithelial subtype.

The Janus kinase 2 is required for expression and nuclear accumulation of cyclin D1 in proliferating mammary epithelial cells.

Using a conditional knockout approach, we previously demonstrated that the Janus kinase 2 (Jak2) is crucial for prolactin (PRL) signaling and normal mammary gland development. PRL is suggested to synchronously activate multiple signaling cascades that emerge on the PRL receptor (PRLR). This study demonstrates that Jak2 is essential for the activation of the signal transducer and activator of transcription 5 (Stat5) and expression of Cish (cytokine-inducible SH2-containing protein), a Stat5-responsive negative regulator of Jak/Stat signaling. However, Jak2 is dispensable for the PRL-induced activation of c-Src, focal adhesion kinase, and the MAPK pathway. Despite activation of these kinases that are commonly associated with proliferative responses, the ablation of Jak2 reduces the multiplication of immortalized mammary epithelial cells (MECs). Our studies show that signaling through Jak2 controls not only the transcriptional activation of the Cyclin D1 gene, but, more importantly, it regulates the accumulation of the Cyclin D1 protein in the nucleus by altering the activity of signal transducers that mediate the phosphorylation and subsequent nuclear export of Cyclin D1. In particular, the levels of activated Akt (protein kinase B) and inactive glycogen synthase kinase-3beta (i.e. a kinase that regulates the nuclear export and degradation of Cyclin D1) are reduced in MECs lacking Jak2. The proliferation of Jak2-deficient MECs can be rescued by expressing of a mutant form of Cyclin D1 that cannot be phosphorylated by glycogen synthase kinase-3beta and therefore constitutively resides in the nucleus. Besides discriminating Jak2-dependent and Jak2-independent signaling events emerging from the PRLR, our observations provide a possible mechanism for phenotypic similarities between Cyclin D1 knockouts and females lacking individual members of the PRLR signaling cascade, in particular the PRLR, Jak2, and Stat5.

Gain-of-function of Stat5 leads to excessive granulopoiesis and lethal extravasation of granulocytes to the lung.

The Signal Transducer and Activator of Transcription 5 (Stat5) plays a significant role in normal hematopoiesis and a variety of hematopoietic malignancies. Deficiency in Stat5 causes impaired cytokine-mediated proliferation and survival of progenitors and their differentiated descendants along major hematopoietic lineages such as erythroid, lymphoid, and myeloid cells. Overexpression and persistent activation of Stat5 are sufficient for neoplastic transformation and development of multi-lineage leukemia in a transplant model. Little is known, however, whether a continuous activation of this signal transducer is essential for the maintenance of hematopoietic malignancies. To address this issue, we developed transgenic mice that express a hyperactive mutant of Stat5 in hematopoietic progenitors and derived lineages in a ligand-controlled manner. In contrast to the transplant model, expression of mutant Stat5 did not adversely affect normal hematopoiesis in the presence of endogenous wildtype Stat5 alleles. However, the gain-of-function of this signal transducer in mice that carry Stat5a/b hypomorphic alleles resulted in abnormally high numbers of circulating granulocytes that caused severe airway obstruction. Downregulation of hyperactive Stat5 in diseased animals restored normal granulopoiesis, which also resulted in a swift clearance of granulocytes from the lung. Moreover, we demonstrate that Stat5 promotes the initiation and maintenance of severe granulophilia in a cell autonomous manner. The results of this study show that the gain-of-function of Stat5 causes excessive granulopoiesis and prolonged survival of granulocytes in circulation. Collectively, our findings underline the critical importance of Stat5 in maintaining a normal balance between myeloid and lymphoid cells during hematopoiesis, and we provide direct evidence for a function of Stat5 in granulophilia-associated pulmonary dysfunction.

Stat5 promotes survival of mammary epithelial cells through transcriptional activation of a distinct promoter in Akt1.

The signal transducer and activator of transcription 5 (Stat5) plays a pivotal role in the proliferation, secretory differentiation, and survival of mammary epithelial cells. However, there is little information about Stat5 target genes that facilitate these biological processes. We provide here experimental evidence that the prolactin-mediated phosphorylation of Stat5 regulates the transcriptional activation of the Akt1 gene. Stat5 binds to consensus sequences within the Akt1 locus in a growth factor-dependent manner to initiate transcription of a unique Akt1 mRNA from a distinct promoter, which is only active in the mammary gland. Elevating the levels of active Akt1 restores the expression of cyclin D1 and proliferation of Jak2-deficient mammary epithelial cells, which provides evidence that Akt1 acts downstream of Jak/Stat signaling. The ligand-inducible expression of Stat5 in transgenic females mediates a sustained upregulation of Akt1 in mammary epithelial cells during the onset of postlactational involution. Stat5-expressing mammary glands exhibit a delay in involution despite induction of proapoptotic signaling events. Collectively, the results of the present study elucidate an underlying mechanism by which active Stat5 mediates evasion from apoptosis and self-sufficiency in growth signals.

What a fish can learn from a mouse: principles and strategies for modeling human cancer in mice.

This review highlights the current techniques used to generate transgenic mouse models of cancer, with an emphasis on recent advances in the use of ubiquitous promoters, models that use Cre-loxP and Flip-FRT recombinase technology, inducible systems, RNAi to target genes, and transposon mutagenesis. A concluding section discusses new imaging systems that visualize tumor progression and the microenvironment in vivo. In this review, these techniques and strategies used in mouse models of cancer are highlighted, as they are pertinent and relevant to the development of zebrafish models of cancer.