Minding the gut: extending embodied cognition and perception to the gut complex.

Scientific and philosophical accounts of cognition and perception have traditionally focused on the brain and external sense organs. The extended view of embodied cognition suggests including other parts of the body in these processes. However, one organ has often been overlooked: the gut. Frequently conceptualized as merely a tube for digesting food, there is much more to the gut than meets the eye. Having its own enteric nervous system, sometimes referred to as the "second brain," the gut is also an immune organ and has a large surface area interacting with gut microbiota. The gut has been shown to play an important role in many physiological processes, and may arguably do so as well in perception and cognition. We argue that proposals of embodied perception and cognition should take into account the role of the "gut complex," which considers the enteric nervous, endocrine, immune, and microbiota systems as well as gut tissue and mucosal structures. The gut complex is an interface between bodily tissues and the "internalized external environment" of the gut lumen, involved in many aspects of organismic activity beyond food intake. We thus extend current embodiment theories and suggest a more inclusive account of how to "mind the gut" in studying cognitive processes.

Protective Microbiota: From Localized to Long-Reaching Co-Immunity.

Resident microbiota do not just shape host immunity, they can also contribute to host protection against pathogens and infectious diseases. Previous reviews of the protective roles of the microbiota have focused exclusively on colonization resistance localized within a microenvironment. This review shows that the protection against pathogens also involves the mitigation of pathogenic impact without eliminating the pathogens (i.e., "disease tolerance") and the containment of microorganisms to prevent pathogenic spread. Protective microorganisms can have an impact beyond their niche, interfering with the entry, establishment, growth, and spread of pathogenic microorganisms. More fundamentally, we propose a series of conceptual clarifications in support of the idea of a "co-immunity," where an organism is protected by both its own immune system and components of its microbiota.

Immune-Mediated Repair: A Matter of Plasticity.

Though the immune system is generally defined as a system of defense, it is increasingly recognized that the immune system also plays a crucial role in tissue repair and its potential dysregulations. In this review, we explore how distinct immune cell types are involved in tissue repair and how they interact in a process that is tightly regulated both spatially and temporally. We insist on the concept of immune cell plasticity which, in recent years, has proved fundamental for the success/understanding of the repair process. Overall, the perspective presented here suggests that the immune system plays a central role in the physiological robustness of the organism, and that cell plasticity contributes to the realization of this robustness.

Impact of cleft width in clefts of secondary palate on the risk of velopharyngeal insufficiency.

Objective To investigate the association between velopharyngeal insufficiency (VPI), a common finding after cleft palate repair, and various risk factors, including cleft width.Methods We performed a retrospective cohort study of patients with isolated cleft palates repaired from 2003 to 2008 at a tertiary children's hospital. Patients were observed postoperatively for development of VPI and palatal fistula. The primary risk factor was cleft width. Covariates included cleft length, age at surgery, and presence of associated syndrome. Logistic regression analysis was used to calculate adjusted and unadjusted odds ratios (ORs).Results The cohort comprised 61 patients. Mean (SD) patient age at the time of cleft repair was 13 (3) months. Fistula rate was 3%. Overall rate of postoperative VPI was 32%. We found significant associations between VPI diagnosis and increasing age in months at the time of palate repair (OR, 1.4 [95% CI, 1.2-1.7]) and between VPI and cleft width greater than 10 mm (OR, 5.3 [95% CI, 1.8-15.6]). The association between VPI and cleft width was similar after adjustment for cleft length, patient age, and syndrome presence (OR, 4.5 [95% CI, 1.1-18.7]).Conclusions Our results suggest that increased palatal cleft width is associated with a greater risk of postoperative VPI. Clinicians should consider this when counseling patient families for cleft palate repair.

Minimizing superficial thermal injury using bilateral cryogen spray cooling during laser reshaping of composite cartilage grafts.

Composite cartilage grafts were excised from New Zealand rabbit ears. Flat composite grafts (of cartilage and overlying skin graft on both surfaces) were obtained from each ear and cut into a rectangle measuring 50 mm by 25 mm (x by y) with an average thickness of approximately 1.3 mm (z), skin included. Specimens were manually deformed with a jig and maintained in this new position during laser illumination. The composite cartilage grafts were illuminated on the concave surface with an Nd:YAG laser (1,064 nm, 3 mm spot) at 10 W, 20 W, 30 W, 40 W, 50 W. Cryogen spray cooling (CSC) was applied to both exterior (convex) and interior (concave) surfaces of the tissue to reduce thermal injury to the grafts. CSC was delivered: (1) in controlled applications (cryogen released when surface reached 40 degrees C, and (2) receiving only laser at above wattage, no CSC [representing the control group]. The specimens were maintained in a deformation for 15 minutes after illumination and serially examined for 14 days. The control group with no CSC caused injury to all specimens, ranging from minor to full thickness epidermal thermal injury. Although most levels of laser and CSC yielded a high degree of reshaping over an acute time period, after 14 days specimens exposed to 30 W, 40 W, 50 W retained shape better than those treated at 10 W and 20 W. The specimens exposed to 50 W with controlled CSC retained its new shape to the highest degree over all others, and thermal injury was minimal. In conclusion, combinations of laser and CSC parameters were effective and practical for the reshaping of composite cartilage grafts.

Using the zebrafish lateral line to screen for ototoxicity.

The zebrafish is a valuable model for studying hair cell development, structure, genetics, and behavior. Zebrafish and other aquatic vertebrates have hair cells on their body surface organized into a sensory system called the lateral line. These hair cells are highly accessible and easily visualized using fluorescent dyes. Morphological and functional similarities to mammalian hair cells of the inner ear make the zebrafish a powerful preparation for studying hair cell toxicity. The ototoxic potential of drugs has historically been uncovered by anecdotal reports that have led to more formal investigation. Currently, no standard screen for ototoxicity exists in drug development. Thus, for the vast majority of Food and Drug Association (FDA)-approved drugs, the ototoxic potential remains unknown. In this study, we used 5-day-old zebrafish larvae to screen a library of 1,040 FDA-approved drugs and bioactives (NINDS Custom Collection II) for ototoxic effects in hair cells of the lateral line. Hair cell nuclei were selectively labeled using a fluorescent vital dye. For the initial screen, fish were exposed to drugs from the library at a 100-muM concentration for 1 h in 96-well tissue culture plates. Hair cell viability was assessed in vivo using fluorescence microscopy. One thousand forty drugs were rapidly screened for ototoxic effects. Seven known ototoxic drugs included in the library, including neomycin and cisplatin, were positively identified using these methods, as proof of concept. Fourteen compounds without previously known ototoxicity were discovered to be selectively toxic to hair cells. Dose-response curves for all 21 ototoxic compounds were determined by quantifying hair cell survival as a function of drug concentration. Dose-response relationships in the mammalian inner ear for two of the compounds without known ototoxicity, pentamidine isethionate and propantheline bromide, were then examined using in vitro preparations of the adult mouse utricle. Significant dose-dependent hair cell loss in the mouse utricle was demonstrated for both compounds. This study represents an important step in validating the use of the zebrafish lateral line as a screening tool for the identification of potentially ototoxic drugs.

Photodynamic therapy on keloid fibroblasts in tissue-engineered keratinocyte-fibroblast co-culture.

BACKGROUND AND OBJECTIVES: Keloids are disfiguring, proliferative scars that are a pathologic response to cutaneous injury. An organotypic tissue culture system (the Raft model 1-10) was used to investigate the feasibility of using photodynamic therapy (PDT) as an adjunctive therapy to treat keloids following surgical excision. The Raft co-culture system mimics skin by layering keratinocytes on top of fibroblasts embedded in a collagen matrix. PDT uses drugs that produce singlet oxygen in situ when irradiated by light, and may lead to a number of effects in living tissues varying from the modulation of growth to apoptosis. PDT is already used to treat several benign and malignant diseases in organs such as the skin, retina, and esophagus. STUDY DESIGN/MATERIALS AND METHODS: Normal adult, neonatal, and keloid fibroblasts and keratinocytes were isolated from skin obtained from patients undergoing elective procedures and used to construct the Rafts. Mature Rafts (after 4 days) were incubated with 5-amino levulinic acid (5-ALA), a photosensitizer, for 3 hours and were laser-irradiated (635 nm) for total energy delivery of 5 J/cm2, 10 J/cm2, or 20 J/cm2. Rafts were examined 24 hours and 14 days later. Cell viability was determined using confocal imaging combined with live-dead fluorescent dyes. Multi-photon microscope (MPM) imaged collagen structure and density. As Rafts contract over time, surface area was measured using optical micrometry daily. RESULTS: At 10 and 20 J/cm2, near-total cell death was observed in all constructs, while at 5 J/cm2 cell viability was comparable to controls. Cell viability in keloid and neonatal Rafts was greater than that observed in normal adult Rafts. Treated Rafts contracted less over the 14-day period compared to controls. Contraction and collagen density were greatest in keloid and neonatal Rafts. CONCLUSIONS: A PDT dosimetry range was established, which reduces tissue contraction and collagen density while minimizing injury to fibroblasts.