Multidisciplinary care improves outcomes for patients with carotid body paragangliomas-The UCLA experience.

Objective: To evaluate the effects and outcomes of multidisciplinary surgical approaches in the management of carotid body tumors (CBT). Methods: A single-center retrospective study at the University of California-Los Angeles Medical Center was conducted on patients who presented with CBTs and underwent surgical resections from 1998 to 2020. Statistical analysis was performed using IBM SPSS v27 and Excel. Results: A total of 75 patients with 79 CBT resections were included. Operating surgical subspecialties included: 41.8% vascular surgery, 24.1% otolaryngology head and neck surgeons (OHNS), and 31.6% combined OHNS and vascular. 68.4% of tumors underwent preoperative embolization. EBL was directly correlated with tumor size. CBT size was similar for OHNS (30 mm) and vascular (31 mm) but was significantly larger for combined OHNS and vascular cases (38 mm). EBL was higher in combined cases (301 mL) compared to OHNS (124 mL) or vascular (203 mL) alone. Incidence of postoperative cranial nerve deficits was 7.8%, with combined OHNS and vascular cases having an incidence of 4.0% when compared to OHNS (5.3%) versus vascular surgery alone (12.1%). Conclusion: CBTs can be managed effectively by single surgical specialties with similar outcomes between vascular surgery and OHNS. In larger, higher grade tumors, however, a combined vascular and OHNS approach had lower incidence of postoperative cranial nerve injuries when compared to single specialty resections, despite a larger EBL. Thus, a multidisciplinary surgical approach suggests favorable outcomes with fewer incidence of cranial nerve deficits for larger, more complex CBT resections. Level of Evidence: 2b-Individual retrospective cohort study.

Single-cell transcriptomics of human-skin-equivalent organoids.

Several methods for generating human-skin-equivalent (HSE) organoid cultures are in use to study skin biology; however, few studies thoroughly characterize these systems. To fill this gap, we use single-cell transcriptomics to compare in vitro HSEs, xenograft HSEs, and in vivo epidermis. By combining differential gene expression, pseudotime analyses, and spatial localization, we reconstruct HSE keratinocyte differentiation trajectories that recapitulate known in vivo epidermal differentiation pathways and show that HSEs contain major in vivo cellular states. However, HSEs also develop unique keratinocyte states, an expanded basal stem cell program, and disrupted terminal differentiation. Cell-cell communication modeling shows aberrant epithelial-to-mesenchymal transition (EMT)-associated signaling pathways that alter upon epidermal growth factor (EGF) supplementation. Last, xenograft HSEs at early time points post transplantation significantly rescue many in vitro deficits while undergoing a hypoxic response that drives an alternative differentiation lineage. This study highlights the strengths and limitations of organoid cultures and identifies areas for potential innovation.

Tissue-engineered vocal fold replacement in swine: Methods for functional and structural analysis.

We have developed a cell-based outer vocal fold replacement (COVR) as a potential therapy to improve voice quality after vocal fold (VF) injury, radiation, or tumor resection. The COVR consists of multipotent human adipose-derived stem cells (hASC) embedded within a three-dimensional fibrin scaffold that resembles vocal fold epithelium and lamina propria layers. Previous work has shown improved wound healing in rabbit studies. In this pilot study in pigs, we sought to develop methods for large animal implantation and phonatory assessment. Feasibility, safety, and structural and functional outcomes of the COVR implant are described. Of eight pigs studied, six animals underwent COVR implantation with harvest between 2 weeks and 6 months. Recovery of laryngeal tissue structure was assessed by vibratory and histologic analyses. Recovery of voice function was assessed by investigating acoustic parameters that were derived specifically for pigs. Results showed improved lamina propria qualities relative to an injured control animal at 6 months. Acoustic parameters reflected voice worsening immediately after surgery as expected; acoustics displayed clear voice recovery in the animal followed for 6 months after COVR. These methods form the basis for a larger-scale long-term pre-clinical safety and efficacy study.

Pharmacists in zoos? A qualitative study investigating the potential for pharmacist involvement in wildlife health care in Aotearoa, New Zealand.

BACKGROUND: The importance of wildlife health has been critically emphasized by the current global pandemic. Pharmacists play a valuable role in the health care of companion animals and livestock; however, their involvement in exotic animal health is largely unexplored. OBJECTIVES: This project consulted with zoo vets in New Zealand and investigated their practices around prescribing and dispensing of medicines to explore the opportunities for the involvement of pharmacists. METHODS: A mixed methods approach was used where data were initially collected through an online survey distributed to 26 veterinarians and animal keepers working in zoos, wildlife parks, and sanctuaries. An optional semistructured interview followed the survey. RESULTS: The facilities surveyed housed New Zealand native animal species and 85% also housed exotic animals. Veterinarians dispensed 75% of medicines at their animal facility, whereas the remaining 25% were dispensed by veterinary nurses. On average, 5-10 medications were dispensed at each animal facility per day. Common medicines dispensed were antibiotics, pain relievers, and antifungals. Most respondents felt that they could benefit from working alongside pharmacists in veterinary care. Compounding, access to medicines and identification of tailored formulations were identified as areas where collaboration would be valued. Limitations in the knowledge of pharmacists in animal medicine were distinguished as an area enhancement to assist in collaborative relationships. CONCLUSIONS: There are opportunities for the skills of pharmacists to be incorporated into the care of animals in zoos and wildlife parks in New Zealand. Strengthening the pharmacist-veterinarian relationship can enhance the health outcomes of animals in animal facilities through this interprofessional interaction.

Objective Assessment of Porcine Voice Acoustics for Laryngeal Surgical Modeling.

Pigs have become important animal models in voice research. Several objective parameters exist to characterize the pig voice, but it is not clear which of them are sensitive to the impaired voice quality after laryngeal injury or surgery. In order to conduct meaningful voice research in pigs, it is critical to have standard functional voice outcome measures that can distinguish between normal and impaired voices. For this reason, we investigated 17 acoustic parameters before and early after surgery in three Yucatan mini pigs. Four parameters showed consistent changes between pre- and post-surgery recordings, mostly related to decreased spectral energy in higher frequencies after surgery. We recommend two of these, 50% spectral energy quartile (Q50) and Flux, for objective functional voice assessment of pigs undergoing laryngeal surgery. The long-term goal of this process is to enable quantitative voice outcome tracking of laryngeal surgical interventions in porcine models.

Ectopic Laryngeal Ossification after Bone Morphogenetic Protein-2.

We report two cases of ectopic bone formation in the head and neck following treatment with recombinant human bone morphogenetic protein-2 (rhBMP-2). Surgical pathologic data, laryngoscopy imaging, CT imaging, and patient medical history were obtained. First, we report osseous metaplasia in the vocal fold in a 67-year-old male following mandibular dental implants with rhBMP-2; second, a case of severe bony overgrowth of the larynx and fusion to the anterior cervical spine (ACS) in a 73-year-old male following multiple anterior cervical discectomies and fusions with rhBMP-2. Ectopic bone formation following rhBMP-2 has been previously reported. Adverse events like local swelling and edema leading to dysphagia and even airway obstruction after cervical spine application of rhBMP-2 have also been widely reported. Due to the uncommon nature of abnormal bony growth in soft tissue areas of the head and neck and the previously documented adverse effects of rhBMP-2 use, especially in the cervical spine, we consider the two unusual case presentations of ectopic bony formation highly likely to be linked with rhBMP-2. We urge awareness of the adverse effects caused by rhBMP-2, and urge caution in dosing.

MP3 Plus: A Modified Micropulse Transscleral Cyclophototherapy Technique for the Treatment of Refractory Glaucoma.

PRéCIS:: In advanced refractory glaucoma which has failed prior MP3 laser, MP3 Plus is effective in reducing intraocular pressure (IOP) without significant complications. It is also useful as a temporizing procedure before incisional surgery. PURPOSE: To evaluate the efficacy and safety of MP3 Plus, a novel technique using modified micropulse transscleral cyclophototherapy (MPTCP) in eyes with refractory glaucoma or failed MPTCP treatment. METHODS: This is a retrospective interventional case series. In total, 32 consecutive eyes from 29 patients with uncontrolled IOPs who had previous MPTCP or refractory glaucoma underwent MP3 Plus, a modified MPTCP where additional discrete pulses are applied. The main outcome measurement was IOP at 1, 3, 6, and 12 months postprocedure, with success defined as a 20% reduction in baseline IOP, an IOP of 25 mmHg or less, and no need for further reoperation. We collected and analyzed visual acuity, number of glaucoma medications, and complications. RESULTS: The patients had an average of 1.4±0.9 glaucoma procedures before treatment, where 87.5% underwent one or more previous MPTCP with suboptimal response. Of the eyes presenting for follow-up, 51.6% (n=31), 37.0% (n=27), 35.7% (n=28), and 25.9% (n=27) achieved the primary outcome at 1, 3, 6, and 12 months, respectively. Baseline IOP was 33.7±11.6 mmHg (n=32 eyes) and posttreatment IOP at 1, 3, 6, and 12 months were 21.0±9.0 mmHg (n=31; P<0.0001), 26.2±10.8 mmHg (n=25; P<0.0001), 23.2±9.4 mmHg (n=20; P<0.05), and 24.6±9.8 mmHg (n=16; P<0.001), respectively. There was a reduction in glaucoma medications from 3.4±0.8 preoperatively to 2.8±1.2 (n=16; P<0.05) at 12 months. There were no cases of prolonged inflammation or hypotony. CONCLUSIONS: MP3 Plus is effective and safe in lowering IOP in eyes with refractory glaucoma after previous failed MPTCP.

Mechanical activation of noncoding-RNA-mediated regulation of disease-associated phenotypes in human cardiomyocytes.

How common polymorphisms in noncoding genome regions can regulate cellular function remains largely unknown. Here we show that cardiac fibrosis, mimicked using a hydrogel with controllable stiffness, affects the regulation of the phenotypes of human cardiomyocytes by a portion of the long noncoding RNA ANRIL, the gene of which is located in the disease-associated 9p21 locus. In a physiological environment, cultured cardiomyocytes derived from induced pluripotent stem cells obtained from patients who are homozygous for cardiovascular-risk alleles (R/R cardiomyocytes) or from healthy individuals who are homozygous for nonrisk alleles contracted synchronously, independently of genotype. After hydrogel stiffening to mimic fibrosis, only the R/R cardiomyocytes exhibited asynchronous contractions. These effects were associated with increased expression of the short ANRIL isoform in R/R cardiomyocytes, which induced a c-Jun N-terminal kinase (JNK) phosphorylation-based mechanism that impaired gap junctions (particularly, loss of connexin-43 expression) following stiffening. Deletion of the risk locus or treatment with a JNK antagonist was sufficient to maintain gap junctions and prevent asynchronous contraction of cardiomyocytes. Our findings suggest that mechanical changes in the microenvironment of cardiomyocytes can activate the regulation of their function by noncoding loci.

Dynamically stiffened matrix promotes malignant transformation of mammary epithelial cells via collective mechanical signaling.

Breast cancer development is associated with increasing tissue stiffness over years. To more accurately mimic the onset of gradual matrix stiffening, which is not feasible with conventional static hydrogels, mammary epithelial cells (MECs) were cultured on methacrylated hyaluronic acid hydrogels whose stiffness can be dynamically modulated from "normal" (<150 Pascals) to "malignant" (>3,000 Pascals) via two-stage polymerization. MECs form and remain as spheroids, but begin to lose epithelial characteristics and gain mesenchymal morphology upon matrix stiffening. However, both the degree of matrix stiffening and culture time before stiffening play important roles in regulating this conversion as, in both cases, a subset of mammary spheroids remained insensitive to local matrix stiffness. This conversion depended neither on colony size nor cell density, and MECs did not exhibit "memory" of prior niche when serially cultured through cycles of compliant and stiff matrices. Instead, the transcription factor Twist1, transforming growth factor ß (TGFß), and YAP activation appeared to modulate stiffness-mediated signaling; when stiffness-mediated signals were blocked, collective MEC phenotypes were reduced in favor of single MECs migrating away from spheroids. These data indicate a more complex interplay of time-dependent stiffness signaling, spheroid structure, and soluble cues that regulates MEC plasticity than suggested by previous models.

A molecular signature for anastasis, recovery from the brink of apoptotic cell death.

During apoptosis, executioner caspase activity has been considered a point of no return. However, recent studies show that cells can survive caspase activation following transient apoptotic stimuli, a process called anastasis. To identify a molecular signature, we performed whole-transcriptome RNA sequencing of untreated, apoptotic, and recovering HeLa cells. We found that anastasis is an active, two-stage program. During the early stage, cells transition from growth-arrested to growing. In the late stage, HeLa cells change from proliferating to migratory. Recovering cells also exhibited prolonged elevation of proangiogenic factors. Strikingly, some early-recovery mRNAs, including Snail, were elevated first during apoptosis, implying that dying cells poise to recover, even while under apoptotic stress. Snail was also required for recovery. This study reveals similarities in the anastasis genes, pathways, and cell behaviors to those activated in wound healing and identifies a repertoire of potential targets for therapeutic manipulation.

Influence of diet on the gut microbiome and implications for human health.

Recent studies have suggested that the intestinal microbiome plays an important role in modulating risk of several chronic diseases, including inflammatory bowel disease, obesity, type 2 diabetes, cardiovascular disease, and cancer. At the same time, it is now understood that diet plays a significant role in shaping the microbiome, with experiments showing that dietary alterations can induce large, temporary microbial shifts within 24 h. Given this association, there may be significant therapeutic utility in altering microbial composition through diet. This review systematically evaluates current data regarding the effects of several common dietary components on intestinal microbiota. We show that consumption of particular types of food produces predictable shifts in existing host bacterial genera. Furthermore, the identity of these bacteria affects host immune and metabolic parameters, with broad implications for human health. Familiarity with these associations will be of tremendous use to the practitioner as well as the patient.