Art, Intuition, and Identity in Ramón y Cajal.

In the history of neuroscience, Cajal stands tall. Many figures in the late 19th and early 20th centuries made major contributions to neuroscience-Sherrington, Ferrier, Jackson, Holmes, Adrian, and Békésy, to name a few. But in the public mind, Cajal is unique. His application of the Golgi method, with an array of histologic stains, unlocked a wealth of new knowledge on the structure and function of the brain. Here we argue that Cajal's success should not only be attributed to the importance of his scientific contributions but also to the artistic visual language that he created and to his pioneering self-branding, which exploited methods of the artist, including classical drawing and the new invention of photography. We argue that Cajal created his distinctive visual language and self-branding strategy by interweaving an ostensibly objective research product with an intimately subjective narrative about the brain and himself. His approach is evident in the use of photography, notably self-portraits, which furthered broad engagement initially inspired by his scientific drawings. Through his visual language, Cajal made an impact in art and culture far beyond the bounds of science, which has sustained his scientific legacy.

Leveraging Collective Impact to Address Structural Racism in Buncombe County, North Carolina.

Buncombe County, North Carolina, was recognized in 2014 as a Robert Wood Johnson Foundation Culture of Health Prize Winner for its work fostering collaboration and partnership to address community health needs. As part of this work, Buncombe County Health and Human Services (HHS) convened a cross-sector Public Health Advisory Council that supported community-based initiatives and ensured that community members were involved in identifying and implementing solutions to issues such as poverty and child well-being. Leveraging existing relationships and past efforts, Buncombe County has continued to build collaborative networks for systems change using a collective impact framework. Bringing together partners across sectors, including the faith community, Black fraternities and sororities, community health workers, consulates, and others, Buncombe County HHS is supporting efforts to train and equip community members to lead health promotion efforts and community conversations on historical trauma and racial healing; engage community members in the policymaking process through town halls; and archive the community's pandemic journey through storytelling. The collective impact framework has shaped an environment that supports community change by centering community aspirations and values. This environment informed recent declarations by Buncombe County HHS and the Board of Commissioners that racism is a public health crisis, as well as a resolution by the city of Asheville supporting community reparations. This article explores how the collective impact framework has been used in Buncombe County to engage and continually invest in communities of color and reviews steps taken to develop and implement an equity action plan to address structural racism.

Clinical Implications of the Submental and Sublingual Arteries in Relation to the Mylohyoid Boutonnière.

OBJECTIVE: Mylohyoid musculature may be included in the submental artery flap to protect perforators. However, blood vessels may pass through the mylohyoid muscle and therefore cause bleeding and risk to pedicle or perforator injury when a mylohyoid-containing flap is lifted. The objectives of this study were to identify the prevalence of the submental and sublingual arteries that traverse the mylohyoid and to assess relationships between vasculature transmitted through mylohyoid muscles and mylohyoid boutonnières. STUDY DESIGN: Cross-sectional human cadaveric study. SETTING: The West Virginia University School of Medicine human gross anatomic laboratories. SUBJECTS AND METHODS: A total of 43 intact mylohyoid muscles from 22 cadavers were dissected. The prevalence of submental vasculature perforating the mylohyoid was recorded in addition to the prevalence and contents of mylohyoid boutonnières. RESULTS: Of 43 mylohyoid muscles, 21 (48.8%) transmitted the submental or sublingual arteries, and 30 (69.1%) possessed boutonnières. One muscle had 2 boutonnières. Of 31 mylohyoid boutonnières, 21 transmitted blood vessels (67.7%). Specifically, 9 transmitted an artery and a vein (29.0%), and 12 transmitted an artery (38.7%). Ten boutonnières (32.3%) were exclusively occupied by fascia. CONCLUSION: This report identifies the importance of identifying and carefully ligating branches of the submental artery that pierce the mylohyoid during elevation of the submental island flap. This report also identifies that a boutonnière is often present where a submental or sublingual artery is traversing the mylohyoid to supply sublingual glands, tongue, and anterior mandible.

Bilateral absence of the musculocutaneous nerve: implications for humerus fracture and atypical median nerve palsy.

The absence of the musculocutaneous nerve represents a failure of the nerve to depart from the median nerve during early development. During a routine dissection of a 66-year-old white female cadaver, a bilateral absence of the musculocutaneous nerve was observed in the upper limbs. Muscles of the anterior flexor compartments of the arms including biceps brachii and brachialis were supplied by branches of the median nerve. The lateral cutaneous nerve of the forearm also branched from the median nerve. In a clinical case of a particularly high median nerve injury, a variation of an absent musculocutaneous nerve may not only result in typical median nerve palsy of the forearm and hand, but palsy in the arm that would manifest as deficiencies in both shoulder and elbow flexion as well as cutaneous sensory loss from the lateral forearm.

An iron triangle ROI model for health care.

OBJECTIVE: Few, if any, return on investment (ROI) analyses of health programs make systematic considerations of patient access, instead focusing principally on gains related to cost and quality. The objective of this study was to develop an open-source model that adds an estimation of gains in patient access to a traditional ROI analysis. A classification system for quantifying gains in patient access is proposed. MATERIALS AND METHODS: An Excel-based ROI model was built that not only incorporated traditional ROI considerations - cost savings and patient cases avoided - but also addressed changes in patient access. The model was then applied in a case study using New Mexico Medicaid data and two proposed initiatives - a statewide health information exchange (HIE) and a community health worker (CHW) program that focused on chronic disease patients. Savings, Health, Outreach, and Access estimates were derived from the literature. ROI estimates were produced that also incorporated relative gains in patient access. RESULTS: Combined, the HIE and CHW programs are predicted to generate a positive ROI by the fourth year, growing to 45% by the program's tenth year. Total estimated cumulative cost for both programs after 10 years is $9,555,226. Total estimated cumulative saving for both programs after 10 years is $11,332,899. Access-related costs begin moderately in year 1 at $122,766 and grow to $1,858,274 by year 10. The model estimates an Access score of 19 in year 1. This figure grew to 380 by year 10. CONCLUSION: Our model shows that a rough estimation of gains in to patient access can be incorporated to traditional ROI analyses. The results of our case study suggest that a CHW program and statewide HIE can generate a positive ROI for the state's Medicaid program.

Ozone exposure initiates a sequential signaling cascade in airways involving interleukin-1beta release, nerve growth factor secretion, and substance P upregulation.

Previous studies demonstrated that interleukin-1ß (IL-1ß) and nerve growth factor (NGF) increase synthesis of substance P (SP) in airway neurons both after ozone (O3) exposure and by direct application. It was postulated that NGF mediates O3-induced IL-1ß effects on SP. The current study specifically focused on the influence of O3 on IL-1ß, NGF, and SP levels in mice bronchoalveolar lavage fluid (BALF) and whether these mediators may be linked in an inflammatory-neuronal cascade in vivo. The findings showed that in vivo O3 exposure induced an increase of all three proteins in mouse BALF and that O3-induced elevations in both NGF and SP are mediated by the inflammatory cytokine IL-1ß. Further, inhibition of NGF reduced O3 induced increases of SP in both the lung BALF and lung tissue, demonstrating NGF serves as a mediator of IL-1ß effects on SP. These data indicate that IL-1ß is an early mediator of O3-induced rise in NGF and subsequent SP release in mice in vivo.

An IMS/ATP Assay for the Detection of Mycobacterium tuberculosis in Urine.

Background. Although sputum smears are the gold standard for diagnosis of tuberculosis, sensitivity in HIV/TB coinfection cases is low, indicating a need for alternative methods. Urine is being increasingly evaluated. Materials and Methods. A novel method for detecting Mycobacterium tuberculosis (MTB) in synthetic urine using a combined IMS/ATP assay was evaluated. Preliminary work established standard ATP conditions and the sensitivity and specificity of the MTB antibody. Eighty-four blinded samples in four replicate assays were evaluated for the presence of MTB using labeled immunomagnetic beads for capture. Beads were separated, washed, and resuspended in broth and added to a microtiter plate. Bioluminescent output was measured and signal-to-noise ratios were calculated. All samples were plated on Middlebrook 7H10 agar or trypticase soy agar to determine limit of detection and recoveries. Results and Conclusions. MTB was distinguished from common bacteriuria isolates and other nontarget bacteria by its ATP results. IMS/ATP successfully detected 19 of 28 samples of MTB in synthetic urine with a limit of detection of 10(4) CFU/ml. Sensitivity and specificity were 67.9% and 82.1%, respectively. This assay offers a possible rapid screening method for HIV-positive patients with suspected coinfection to improve MTB diagnosis.

Early Postnatal Ozone Exposure Alters Rat Nodose and Jugular Sensory Neuron Development.

Sensory neurons originating in nodose and jugular ganglia that innervate airway epithelium (airway neurons) play a role in inflammation observed following exposure to inhaled environmental irritants such as ozone (O(3)). Airway neurons can mediate airway inflammation through release of the neuropeptide substance P (SP). While susceptibility to airway irritants is increased in early life, the developmental dynamics of afferent airway neurons are not well characterized. The hypothesis of this study was that airway neuron number might increase with increasing age, and that an acute, early postnatal O(3) exposure might increase both the number of sensory airway neurons as well as the number SP-containing airway neurons. Studies using Fischer 344 rat pups were conducted to determine if age or acute O(3) exposure might alter airway neuron number. Airway neurons in nodose and jugular ganglia were retrogradely labeled, removed, dissociated, and counted by means of a novel technique employing flow cytometry. In Study 1, neuron counts were conducted on postnatal days (PD) 6, 10, 15, 21, and 28. Numbers of total and airway neurons increased significantly between PD6 and PD10, then generally stabilized. In Study 2, animals were exposed to O(3) (2 ppm) or filtered air (FA) on PD5 and neurons were counted on PD10, 15, 21, and 28. O(3) exposed animals displayed significantly less total neurons on PD21 than FA controls. This study shows that age-related changes in neuron number occur, and that an acute, early postnatal O(3) exposure significantly alters sensory neuron development.

Dead-end ultrafiltration concentration and IMS/ATP-bioluminescence detection of Escherichia coli O157:H7 in recreational water and produce wash.

The purpose of this study was to develop a detection method for viable E. coli O157:H7 in fresh produce and recreational water. The method was evaluated using eight samples of produce wash and recreational water with or without spiked E. coli O157:H7 at ≤10(2) CFU·ml(-1) and concentrated using dead-end ultrafiltration (DEUF) to produce primary and secondary retentates. Fifty-four matrix replicates of undiluted secondary retentates or dilutions (1:2 or 1:10 in buffer) were evaluated using an IMS/ATP bioluminescence assay (IMS/ATP). Combining primary and secondary DEUF yielded a 2-4 log(10) increase in E. coli O157:H7 concentrations in spiked samples and resulted in signal-to-noise ratios 2-219 fold higher than controls, depending on the sample type. DEUF increased the concentration of E. coli O157:H7 to within the detectable limits of IMS/ATP. The combined assay provided detection of viable E. coli O157:H7 in produce and recreational water. Accurate detection of microbial pathogens using DEUF and IMS/ATP could reduce disease outbreaks from contaminated water sources and food products.

Role of nerve growth factor in ozone-induced neural responses in early postnatal airway development.

Airway neural plasticity contributes to the process of airway remodeling in response to airway irritants. However, the mechanisms of neural remodeling in the airways during the early postnatal period, when responses to airway irritation may be most sensitive, have not been characterized. This study used a rat model to examine a possible mechanism of ozone (O(3))-induced neural hyperresponsiveness during a critical period of developmental, postnatal day (PD) 6, that may be mediated by the neurotrophin nerve growth factor (NGF), resulting in an enhanced release of inflammatory neuropeptide substance P (SP) from airway nerves. Rat pups between PD6-PD28 were killed 24 hours after exposure to O(3) (2 ppm, 3 hours) or filtered air (FA), to establish a timeline of NGF synthesis, or else they were exposed to O(3) or NGF on PD6 or PD21 and re-exposed to O(3) on PD28, and killed on PD29. Measurement endpoints included NGF mRNA in tracheal epithelial cells, NGF protein in bronchoalveolar lavage fluid, airway SP-nerve fiber density (NFD), and SP-positive airway neurons in vagal ganglia. Acute exposure to O(3) increased NGF in bronchoalveolar lavage fluid on PD10 and PD15, and mRNA expression in epithelial cells on PD6, compared with FA controls. NGF protein and mRNA expression in the O(3)-PD6/O(3)-PD28 groups were significantly higher than in the O(3)-PD21/O(3)-PD28 and O(3)-PD6/FA-PD28 groups. NGF-PD6/O(3)-PD28 increased the SP innervation of airway smooth muscle and SP-positive sensory neurons, compared with the NGF-PD21/O(3)-PD28 or NGF-PD6/FA-PD28 groups. NGF enhanced sensory innervation, which may mediate acute responses or prolong sensitivity to O(3) during early life. The model may be relevant in O(3) responses during early childhood.

Rapid detection and identification of bacterial pathogens by using an ATP bioluminescence immunoassay.

Rapid identification of viable bacterial contaminants in food products is important because of their potential to cause disease. This study examined a method for microbial detection by using a combined ATP bioluminescence immunoassay. Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium were selected as target organisms because of their implication in foodborne illness. Various matrices containing the target cells were examined, including ground beef homogenate, apple juice, milk, and phosphate-buffered saline. Specific antibodies were immobilized on the surface of 96-well plates, and then the sample matrices containing target cells in the wells were incubated. Sample matrix (no cells) was used to establish background. The plates were washed, and the wells were incubated with BacTiter-Glo reagent in Mueller-Hinton II broth. Bioluminescent output was measured with the GloMax 96 luminometer. Signal-to-noise ratios were calculated, resulting in a limit of detection of 10(4) CFU/ml for both E. coli O157:H7 and Salmonella Typhimurium. The limit of detection for both species was not affected by the presence of nontarget cells. The various sample matrices did not affect signal-to-noise ratios when E. coli O157:H7 was the target. A weak matrix effect was observed when Salmonella Typhimurium was the target. A strong linear correlation was observed between the number of cells and luminescent output over 4 orders of magnitude for both species. This method provides a means of simultaneously detecting and identifying viable pathogens in complex matrices, and could have wider application in food microbiology.

Sensory neural responses to ozone exposure during early postnatal development in rat airways.

Airway infections or irritant exposures during early postnatal periods may contribute to the onset of childhood asthma. The purpose of this study was to examine critical periods of postnatal airway development during which ozone (O(3)) exposure leads to heightened neural responses. Rats were exposed to O(3) (2 ppm) or filtered air for 1 hour on specific postnatal days (PDs) between PD1 and PD29, and killed 24 hours after exposure. In a second experiment, rats were exposed to O(3) on PD2-PD6, inside a proposed critical period of development, or on PD19-PD23, outside the critical period. Both groups were re-exposed to O(3) on PD28, and killed 24 hours later. Airways were removed, fixed, and prepared for substance P (SP) immunocytochemistry. SP nerve fiber density (NFD) in control extrapulmonary (EXP) epithelium/lamina propria (EPLP) increased threefold, from 1% to 3.3% from PD1-PD3 through PD13-PD15, and maintained through PD29. Upon O(3) exposure, SP-NFD in EXP-smooth muscle (SM) and intrapulmonary (INT)-SM increased at least twofold at PD1-PD3 through PD13-PD15 in comparison to air exposure. No change was observed at PD21-PD22 or PD28-PD29. In critical period studies, SP-NFD in the INT-SM and EXP-SM of the PD2-PD6 O(3) group re-exposed to O(3) on PD28 was significantly higher than that of the group exposed at PD19-PD23 and re-exposed at PD28. These findings suggest that O(3)-mediated changes in sensory innervation of SM are more responsive during earlier postnatal development. Enhanced responsiveness of airway sensory nerves may be a contributing mechanism of increased susceptibility to environmental exposures observed in human infants and children.

Prenatal and early, but not late, postnatal exposure of mice to sidestream tobacco smoke increases airway hyperresponsiveness later in life.

BACKGROUND: Cigarette smoke exposure in utero and during early postnatal development increases the incidence of asthma and airway hyperresponsiveness (AHR) later in life, suggesting that a possible critical period of developmental sensitivity exists in the prenatal and early postnatal periods. OBJECTIVE: We investigated mechanisms of susceptibility during critical developmental periods to sidestream smoke (SS) exposure and evaluated the possible effects of SS on neural responses. METHODS: We exposed three different age groups of mice to either SS or filtered air (FA) for 10 consecutive days beginning on gestation day (GD) 7 by maternal exposure or beginning on postnatal day (PND) 2 or PND21 by direct inhalation. Lung function, airway substance P (SP) innervation, and nerve growth factor (NGF) levels in broncho alveolar lavage fluid were measured after a single SS exposure on PND59. RESULTS: Methacholine (MCh) dose response for lung resistance (R(L)) was significantly elevated, and dynamic pulmonary compliance (C(dyn)) was significantly decreased, in the GD7 and PND2 SS exposure groups compared with the FA groups after SS exposure on PND59. At the same time points, the percent area of SP nerve fibers in tracheal smooth muscle and the levels of NGF were significantly elevated. MCh dose-response curves for R(L) and C(dyn), SP nerve fiber density, and the level of NGF were not significantly changed in the PND21 exposure group after SS exposure on PND59. CONCLUSIONS: These results suggest that a critical period of susceptibility to SS exposure exists in the prenatal and early postnatal period of development in mice that results in increased SP innervation, increased NGF levels in the airway, and enhanced MCh AHR later in life.

Allergen-induced substance P synthesis in large-diameter sensory neurons innervating the lungs.

BACKGROUND: Tachykinins such as substance P are localized in unmyelinated slow-conducting C fibers that can be activated by noxious stimuli and tissue inflammation. Substance P is seldom expressed in fast-conducting large-diameter (A-fiber) vagal sensory neurons. We have previously found that allergic inflammation causes a phenotypic change in tachykinergic innervation of the trachea such that the production of substance P is induced in large-diameter sensory neurons projecting mechanosensitive A fibers to the trachea. OBJECTIVE: To evaluate whether allergic inflammation also induces substance P synthesis in large-diameter sensory stretch-receptor neurons innervating guinea pig lungs, and to investigate potential mechanisms by which this may occur. METHODS: Sensitized guinea pigs were exposed to allergen (ovalbumin) aerosol. One day later, immunohistochemical analysis was performed on vagal sensory neurons that had been retrogradely labeled from the lungs. RESULTS: Ovalbumin inhalation caused a significant increase in substance P expression in large-diameter neurofilament-positive nodose ganglion neurons that innervate the lungs (P < .05). This effect was decreased by ipsilateral vagotomy. Exposing isolated nodose ganglia to the sensitizing antigen, ovalbumin, also significantly increased substance P expression compared with control. CONCLUSION: Allergic inflammation induces substance P synthesis in large-diameter (A-fiber) nodose ganglion neurons innervating guinea pig lungs. This could contribute to the hyperreflexia seen in allergic airway disease. The full expression of this phenotypic switch in vagus nodose ganglion neurons requires intact vagus nerve, but if allergen reached the systemic circulation in sufficient quantities, it could also affect substance P synthesis by local activation of vagal ganglionic mast cells.

Expression of tachykinins in nonnociceptive vagal afferent neurons during respiratory viral infection in guinea pigs.

Immunohistochemistry was combined with retrograde labeling to characterize the effect of respiratory infection with Sendai virus on the number of Substance P/Neurokinin A-containing vagal afferent neurons whose cell bodies resided in the nodose ganglia and whose receptive fields were located in guinea pig trachea. Of the neurons labeled from the trachea of vehicle-inoculated guinea pigs, few stained positively for Substance P/Neurokinin A (approximately 3% of total labeled neurons). These neurons had small diameter cell bodies (mode = 16-20 microm), a feature of nociceptive-like C-fibers. Viral infection (Day 4 after inoculation) was associated with a significantly greater number of labeled neurons containing Substance P/Neurokinin A (approximately 20% of total labeled neurons). The majority of these had a relatively large cell body diameter (mode = 36- 40 microm), a feature of nonnociceptive afferent neurons. This induction appeared to be reversible as there were significantly fewer Substance P/Neurokinin A positive neurons in nodose ganglia from virus-inoculated guinea pigs at Day 28 after inoculation, a time point when virus-induced airway inflammation had all but resolved. These findings support the hypothesis that viral infection leads to a qualitative change in the vagal afferent innervation of guinea pig airways such that both small diameter nociceptive-like neurons and large diameter nonnociceptive neurons express tachykinins.