Corrigendum to "Single-chain fragment variable antibody targeting cholecystokinin-B receptor for pain reduction" [Neurobiol. Pain 10 (2021) 100067].

[This corrects the article DOI: 10.1016/j.ynpai.2021.100067.].

Evaluation of daily environmental cleaning and disinfection practices in veterans affairs acute and long-term care facilities: A mixed methods study.

OBJECTIVES: To describe daily environmental cleaning and disinfection practices and their associations with cleaning rates while exploring contextual factors experienced by healthcare workers involved in the cleaning process. METHODS: A convergent mixed methods approach using quantitative observations (ie, direct observation of environmental service staff performing environmental cleaning using a standardized observation form) and qualitative interviews (ie, semistructured interviews of key healthcare workers) across 3 Veterans Affairs acute and long-term care facilities. RESULTS: Between December 2018 and May 2019 a total of sixty-two room observations (N = 3602 surfaces) were conducted. The average observed surface cleaning rate during daily cleaning in patient rooms was 33.6% for all environmental surfaces and 60.0% for high-touch surfaces (HTS). Higher cleaning rates were observed with bathroom surfaces (Odds Ratio OR = 3.23), HTSs (OR = 1.57), and reusable medical equipment (RME) (OR = 1.40). Lower cleaning rates were observed when cleaning semiprivate rooms (OR = 0.71) and rooms in AC (OR = 0.56). In analysis stratified by patient presence (ie, present, or absent) in the room during cleaning, patient absence was associated with higher cleaning rates for HTSs (OR = 1.71). In addition, the odds that bathroom surfaces being cleaned more frequently than bedroom surfaces decreased (OR = 1.97) as well as the odds that private rooms being cleaned more frequently than semi-private rooms also decreased (OR = 0.26; 0.07-0.93). Between January and June 2019 eighteen qualitative interviews were conducted and found key themes (ie, patient presence and semiprivate rooms) as potential barriers to cleaning; this supports findings from the quantitative analysis. CONCLUSION: Overall observed rates of daily cleaning of environmental surfaces in both acute and long-term care was low. Standardized environmental cleaning practices to address known barriers, specifically cleaning practices when patients are present in rooms and semi-private rooms are needed to achieve improvements in cleaning rates.

Single-chain Fragment variable antibody targeting cholecystokinin-B receptor for pain reduction.

The cholecystokinin B receptor and its neuropeptide ligand are upregulated in chronic neuropathic pain models. Single-chain Fragment variable antibodies were generated as preferred non-opioid targeting therapy blocking the cholecystokinin B receptor to inhibit chronic neuropathic pain models in vivo and in vitro. Engineered antibodies of this type feature binding activity similar to monoclonal antibodies but with stronger affinity and increased tissue penetrability due to their smaller size. More importantly, single-chain Fragment variable antibodies have promising biotherapeutic applications for both nervous and immune systems, now recognized as interactive in chronic pain. A mouse single-chain Fragment variable antibody library recognizing a fifteen amino acid extracellular peptide fragment of the cholecystokinin B receptor was generated from immunized spleens. Ribosome display, a powerful cell-free technology, was applied for recombinant antibody selection. Antibodies with higher affinity, stability, solubility, and binding specificity for cholecystokinin B not A receptor were selected and optimized for in vivo and in vitro efficacy. A single dose of the lead candidate reduced mechanical and cold hypersensitivity in two rodent models of neuropathic pain for at least seven weeks. Continuing efficacy was evident with either intraperitoneal or intranasal dosing. Likewise, the lead single-chain Fragment variable antibody totally prevented development of anxiety- and depression-like behaviors and cognitive deficits typical in the models. Reduction of neuronal firing frequency was evident in trigeminal ganglia primary neuronal cultures treated in vitro with the cholecystokinin B receptor antibody. Immunofluorescent staining intensity in the trigeminal neuron primary cultures was significantly reduced incrementally after overnight binding with increasingly higher dilutions of the single-chain Fragment variable antibody. While it is reported that single-chain Fragment variable antibodies are removed systemically within 2-6 h, Western blot evidence indicates the His-tag marker remained after 7 weeks in the trigeminal ganglia and in the dorsolateral medulla, providing evidence of brain and ganglia penetrance known to be compromised in overactivated states. This project showcases the in vivo efficacy of our lead single-chain Fragment variable antibody indicating its potential for development as a non-opioid, non-addictive therapeutic intervention for chronic pain. Importantly, studies by others have indicated treatments with cholecystokinin B receptor antagonists suppress maintenance and reactivation of morphine dependence in place preference tests while lowering tolerance and dose requirements. Our future studies remain to address these potential benefits that may accompany the cholecystokinin B receptor biological therapy. Both chronic sciatic and orofacial pain can be unrelenting and excruciating, reducing quality of life as well as diminishing physical and mental function. An effective non-opiate, non-addictive therapy with potential to significantly reduce chronic neuropathic pain long term is greatly needed.

Performance of endoscopic repair with endonasal flaps for congenital choanal atresia. A systematic review / Rendimiento de reparación endoscópica de atresia de coanas con colgajos endonasales. Revisión sistemática

Endoscopic repair of congenital choanal atresia is the gold standard surgical treatment today. Though several controversies on treatment have been reported, surgical techniques for better outcomes are still in discussion. The objective of this study is to evaluate the performance of endoscopic choanal atresia repair with endonasal flaps and no stents. Publications in English in the last 5 years were searched in the PUBMED database and were systematically reviewed. A total of 9 articles were included according to the inclusion criteria, obtaining a total of 266 patients managed for congenital choanal atresia with endoscopic surgery, endonasal flaps, and no stents. Surgical results, type of atresia, atresia laterality, associated pathologies and follow up were evaluated. Successful surgery was obtained in 237 (89%) patients while 29 (11%) patients required a new surgical intervention during the follow-up period. Fourteen percent of the patients were diagnosed with CHARGE syndrome and 5% of the patients had some associated heart disease. Bony-Membranous stenosis was observed in 74% of the patients, while a total bony obstruction was recognized in 26% of the patients. Unilateral atresia was observed in 37% of the cases and 63% of the cases had bilateral atresia. The mean follow-up period was 39.5 months (range 3-168 months). An important functional success rate can be accomplished by correcting congenital choanal atresia using functional endoscopic surgery, covering raw areas with endonasal vascularized flaps, avoiding postoperative endonasal stenting

La reparación endoscópica de la atresia de coanas es hoy en día el tratamiento de elección. Existen controversias con respecto a la técnica quirúrgica que aporte los mejores resultados. El objetivo de este estudio es evaluar el rendimiento de la reparación endoscópica de la atresia de coanas con uso de colgajos intranasales, sin uso de stents. Se realizó una revisión sistemática de los artículos escritos en inglés publicados en la base de datos de PUBMED en los últimos 5 años. Un total de 9 artículos cumplieron los criterios de inclusión, reuniendo 266 pacientes que fueron tratados de una atresia de coanas con técnica endoscópica con colgajos intranasales, sin uso de stents. Las variables evaluadas fueron: el tipo de atresia, los resultados quirúrgicos, la lateralidad, la enfermedad asociada y el seguimiento. En 237 pacientes (89%) se consiguió un resultado satisfactorio, mientras que 29 pacientes (11%) requirieron una nueva intervención quirúrgica durante el seguimiento. El 14% de los pacientes fueron diagnosticados de síndrome de CHARGE y un 5% tuvieron alguna cardiopatía asociada. Una estenosis óseo-membranosa fue encontrada en un 74%, mientras una estenosis totalmente ósea fue observada en un 26%. Lesiones unilaterales fueron observadas en el 37% de los casos y bilaterales en el 63% de los casos. El seguimiento medio fue de 39,5 meses (rango: 3-168 meses). Un éxito quirúrgico funcional se puede obtener usando cirugía endoscópica nasal, cubriendo las zonas de exposición ósea con colgajos nasales vascularizados y evitando el uso de stents postoperatorios

Performance of endoscopic repair with endonasal flaps for congenital choanal atresia. A systematic review.

Endoscopic repair of congenital choanal atresia is the gold standard surgical treatment today. Though several controversies on treatment have been reported, surgical techniques for better outcomes are still in discussion. The objective of this study is to evaluate the performance of endoscopic choanal atresia repair with endonasal flaps and no stents. Publications in English in the last 5 years were searched in the PUBMED database and were systematically reviewed. A total of 9 articles were included according to the inclusion criteria, obtaining a total of 266 patients managed for congenital choanal atresia with endoscopic surgery, endonasal flaps, and no stents. Surgical results, type of atresia, atresia laterality, associated pathologies and follow up were evaluated. Successful surgery was obtained in 237 (89%) patients while 29 (11%) patients required a new surgical intervention during the follow-up period. Fourteen percent of the patients were diagnosed with CHARGE syndrome and 5% of the patients had some associated heart disease. Bony-Membranous stenosis was observed in 74% of the patients, while a total bony obstruction was recognized in 26% of the patients. Unilateral atresia was observed in 37% of the cases and 63% of the cases had bilateral atresia. The mean follow-up period was 39.5 months (range 3-168 months). An important functional success rate can be accomplished by correcting congenital choanal atresia using functional endoscopic surgery, covering raw areas with endonasal vascularized flaps, avoiding postoperative endonasal stenting.

Transcanal endoscopic type 1 tympanoplasty in children: Cartilage butterfly and fascia temporalis graft.

INTRODUCTION: Endoscopic type 1 tympanoplasty is every day gaining numerous adepts for tympanic membrane repair. Due to the value of reducing postauricular approaches, decreasing postoperative morbidity and hospitalization time. The objective of this study is to present surgical results of endoscopic type 1 tympanoplasty in the pediatric population using fascia temporalis or cartilage butterfly graft. MATERIALS AND METHODS: Prospective study regarding the pediatric population, mean age of 10.7 years old. Patients diagnosed with chronic otitis media without cholesteatoma and intact ossicular chain. Tympanic membrane reconstruction using inlay cartilage butterfly graft or underlay fascia temporalis graft according to surgical needs. Audiograms were evaluated preoperatively and 6 months after surgery. No postauricular approaches were performed. RESULTS: A total of 54 ears were operated, 25 utilizing underlay fascia temporalis graft and 29 using inlay cartilage butterfly graft. Six months following surgery, dry and closed tympanic membranes were obtained in 54 cases (92.6%). Preoperative and postoperative air conduction (AC) thresholds, bone conduction (BC) thresholds and air-bone gaps (ABG) were assessed. Preoperative AC of 24.6 dB, BC of 8.9 dB and an ABG of 15.5 dB. Postoperative AC of 16.3 dB, BC of 8.9 and an ABG of 6.9 dB. A postoperative ABG reduction of 8.5 dB was reached. CONCLUSION: Transcanal endoscopic type 1 tympanoplasty can be achieved in every pediatric patient with chronic otitis media without cholesteatoma, and, is a safe and efficient procedure.

Bone repair cells for craniofacial regeneration.

Reconstruction of complex craniofacial deformities is a clinical challenge in situations of injury, congenital defects or disease. The use of cell-based therapies represents one of the most advanced methods for enhancing the regenerative response for craniofacial wound healing. Both somatic and stem cells have been adopted in the treatment of complex osseous defects and advances have been made in finding the most adequate scaffold for the delivery of cell therapies in human regenerative medicine. As an example of such approaches for clinical application for craniofacial regeneration, Ixmyelocel-T or bone repair cells are a source of bone marrow derived stem and progenitor cells. They are produced through the use of single pass perfusion bioreactors for CD90+ mesenchymal stem cells and CD14+ monocyte/macrophage progenitor cells. The application of ixmyelocel-T has shown potential in the regeneration of muscular, vascular, nervous and osseous tissue. The purpose of this manuscript is to highlight cell therapies used to repair bony and soft tissue defects in the oral and craniofacial complex. The field at this point remains at an early stage, however this review will provide insights into the progress being made using cell therapies for eventual development into clinical practice.

Development and characterization of an in vitro gingival epithelial model.

A 3-dimensional gingival epithelial model has been developed and characterized. Oral epithelial cells and connective tissue fibroblasts were isolated from human gingival tissue and used to create an in vitro oral mucosa co-culture model. Fibroblasts were seeded on a scaffold of nylon mesh, allowed to proliferate and secrete collagen and extracellular matrix proteins to form a stroma capable of supporting the growth of epithelial cells. Epithelial cells were seeded on top of a confluent stromal layer, proliferated and differentiated to form a stratified squamous epithelium. Resident epithelial cells were stimulated, by manipulation of growth medium and culture conditions, to form a multi-layered oral mucosa-like tissue. Histologic analyses revealed cellular architecture exhibiting stromal-epithelial interaction which supports the growth and differentiation of an epithelial layer. Immunohistochemical analyses confirmed production of types I and III collagen. Immunofluorescence of the stromal layer identified type IV collagen and fibronectin. Fibronectin was also detected on surface epithelium. Differentiation of basal, spinous and granular cells was observed, and the presence of differentiation markers, acidic (K10, 14-16, 19) and basic (K1-8) cytokeratins were confirmed using broad spectrum cytokeratin antibodies, AE1 and AE3. Development of a discontinuous basal lamina zone, with hemidesmosomes, was observed by electron microscopy. The co-culture was metabolically active, as measured by the thiazoyl blue (MTT) assay for mitchondrial function and [3H] thymidine incorporation into DNA. The human gingival epithelial co-culture model was viable up to 35 days post-epithelial seed. This model may offer opportunities for limited study of periodontal tissue responsiveness.

Comparison of measures used to determine substance abuse in an inpatient psychiatric population.

Eighty-nine of 178 consecutively admitted inpatients were administered the substance abuse sections of the Structured Clinical Interview for DSM-III-R--Patient Version (SCID-P). Patients also provided a urine sample for toxicologic analysis. In addition, primary clinicians assigned admission and discharge diagnoses. Test characteristics (e.g., sensitivity) for confirming a current diagnosis of psychoactive substance abuse were calculated for each measure and compared. Urine toxicology analyses and admission and discharge diagnoses were significantly less accurate in diagnosing psychoactive substance abuse than the SCID-P. This finding indicates that substance abuse is frequently not noted during routine admission and discharge assessments.

Composite grafts of human keratinocytes grown on a polyglactin mesh-cultured fibroblast dermal substitute function as a bilayer skin replacement in full-thickness wounds on athymic mice.

We have developed and tested in athymic mice a new, cultured, dermal-epidermal graft composed of two human cell types coupled with a biodegradable dermal scaffold. Cultured, proliferating human keratinocytes (HK) were applied to the surface of a living dermal tissue replacement that is composed of human fibroblasts cultured on a polyglactin mesh. After 4 to 6 days of coculture, proliferating HKs achieved confluency on the surface of the living dermal tissue replacement. Grafts were then transferred to full-thickness wounds on the dorsum of athymic mice. Sixteen animals were grafted, and the mean percentage of graft take (original wound area covered) on day 20 after grafting was 51.25%. Staining with antibody specific for human involucrin confirmed the presence of HKs on closed wounds, and staining with antibody specific for human laminin revealed a continuous layer of laminin at the dermal-epidermal junction on day 20. Animals closed with living dermal tissue replacement alone markedly contracted, whereas application of living dermal tissue replacement-HK grafts appeared to retard contraction. Because polyglactin mesh fibers are absorbed by hydrolysis rather than by enzymatic degradation, this living composite graft may be more resistant to destruction when placed on excised human wounds than are composite grafts, which are composed of a collagen matrix. The inclusion of the living dermal substitute may ultimately provide better skin quality than is achieved from the use of cultured keratinocytes alone. Fragility of the epidermal layer is probably due to the short-term culture of HKs on the living dermal tissue replacement, and further efforts to develop a thicker epithelial layer may improve graft durability.

Culturing keratinocytes and fibroblasts in a three-dimensional mesh results in epidermal differentiation and formation of a basal lamina-anchoring zone.

The purpose of this study was to characterize an in vitro co-culture model in which fibroblasts grown in a three-dimensional nylon mesh were recombined with human keratinocytes. The cultures were kept for 3 and 5 weeks and then processed for electron microscopy. Keratinocytes showed reconstruction of an epidermis consisting of a basal layer with hemidesmosomes, a stratified epithelium with tonofilaments and desmosomes, a granular layer with keratinosomes and keratohyaline granules, and a transitional stratum corneum. Anchoring filaments, lamina densa, anchoring fibrils, bundles of elastin-associated microfibrils (diameters 10 nm) and fine collagen fibrils were formed. Collagen fibrils near the epidermis were much thinner than those in the lower levels. The present study shows that the dermal model containing metabolically active fibroblasts in their natural environment will support epidermal morphogenesis and differentiation including the formation of a basal lamina and anchoring zone.

Characterization, barrier function, and drug metabolism of an in vitro skin model.

We have characterized an in vitro skin model consisting of neonatal keratinocytes and fibroblasts grown on a nylon mesh. To produce a dermal model, fibroblasts were seeded onto nylon mesh and grown for 4 weeks until a physiologic dermal-like matrix was formed. This matrix was found to consist of collagens I and III, fibronectin, and glycosaminoglycans. Keratinocytes were then seeded onto the dermal model and the co-culture was grown at the air/liquid interface. A differentiated epidermis with distinct basal, spinous, granular, and stratum corneum layers was formed. When incubated in the presence of keratinocytes, fibronectin immunofluorescence increased throughout the dermis compared to cultures incubated similarly in the absence of keratinocytes. A basement membrane zone rich in laminin, collagen IV, and heparan sulfate proteoglycan was detected. The epidermis, isolated from the co-culture by thermolysin digestion, was analyzed for differentiation markers. K1 keratin (67-kDa) and involucrin were detected by immunologic techniques. Ceramide lipids (types III and IV), thought to be important in barrier function, were detected by thin-layer chromatography. The permeability of the co-culture to a panel of compounds, including [3H]-water, was determined using Franz and side-by-side diffusion cells. The permeability coefficient for water was of the same order of magnitude as that determined for neonatal foreskin. The co-culture also showed selective permeability to a panel of compounds of differing lipid solubility. This co-culture metabolized [3H]-testosterone to a profile of metabolites similar to that of neonatal foreskin. We believe that this in vitro skin model will be useful for the study of drug permeability and metabolism.

Evaluation of a biodegradable matrix containing cultured human fibroblasts as a dermal replacement beneath meshed skin grafts on athymic mice.

Meshed, expanded split-thickness skin grafts (MSTSG) frequently achieve poor results when used to cover full-thickness wounds. Poor cosmetic and functional results occur in part because the epithelium that grows across the skin graft interstices lacks a dermis. We used a living dermal replacement composed of either polyglycolic acid (PGA) or polyglactin-910 (PGL) mesh containing confluent, cultured human fibroblasts. These grafts were applied to full-thickness wounds on athymic mice; widely expanded, 3:1 ratio human MSTSG was then placed over the dermal graft. Histologic examination of wounds during a 99-day period after graft placement showed that PGA/PGL-fibroblast grafts vascularized to the wound, and the MSTSG simultaneously vascularized to the PGA/PGL-fibroblast graft. Epithelialization from the MSTSG bridges proceeded rapidly across the surface of the PGA/PGL-fibroblast grafts, resulting in an epithelialized layer that covered a densely cellular substratum that resembled dermis. Basement membrane formation at the dermal-epidermal junction of the epithelialized interstices was confirmed by immunohistochemical microscopy. Minimal inflammatory reaction to the PGA/PGL-fibroblast grafts was seen. Grafts composed of PGA or PGL biodegradable meshes combined with cultured fibroblasts vascularize in full-thickness wounds, resulting in formation of organized tissue beneath the epithelialized surface that resembles dermis.

In vivo optimization of a living dermal substitute employing cultured human fibroblasts on a biodegradable polyglycolic acid or polyglactin mesh.

The design of a skin-substitute must address the need for a dermal component, as this mesenchymally-derived tissue is important in maintaining the integrity and function of skin. An in vivo study was undertaken to assess the use of two biodegradable meshes, polyglycolic acid and polyglactin-910, as carriers for cultured human fibroblasts in a living dermal replacement. The consistent vascularization and epithelialization of these grafts placed on athymic mice showed that this has potential in re-creating the dermis in a skin-substitute.

The use of cultured epithelial and endothelial cells for drug transport and metabolism studies.

In an effort to develop novel strategies for delivery of drug candidates arising from rational drug design and recombinant DNA technology, pharmaceutical scientists have begun to employ the techniques of cell culture to study drug transport and metabolism at specific biological barriers. This review describes some of the general factors that should be considered in developing a cell culture model for transport studies and metabolism studies. In addition, we review in detail the recent progress that has been made in establishing, validating, and using cell cultures of epithelial barriers (e.g., cells that constitute the intestinal, rectal, buccal, sublingual, nasal, and ophthalmic mucosa as well as the epidermis of the skin) and the endothelial barriers (e.g., brain microvessel endothelial cells).

Part II: Sex differences in persons found not guilty by reason of insanity: analysis of data from the Connecticut NGRI Registry.

Thirty-one female insanity acquittess from Connecticut were matched to a group of 31 male NGRIs. The samples were compared with regard to demographic, criminal, and clinical characteristics. Logistic regression analyses were used to determine predictors of criminal recidivism for the sample. Results indicated that women NGRIs were older, more likely to be married, less likely to be substance abusers, had less extensive criminal records, and were released from hospitals sooner than the men. A significant racial difference was noted: white women had less extensive criminal records and were hospitalized for shorter periods than minority women. Results of the logistic regression analyses showed that the strongest independent predictors of criminal recidivism were race and having a diagnosis other than psychosis (schizophrenia, affective or organic disorders). Findings support recent APA policy guidelines on the insanity defense.

An analysis of the scholarly activities of respiratory care faculty in southern academic health centers.

UNLABELLED: Little is known about the scholarly production of faculty members who teach in respiratory care programs. METHOD: We studied the scholarly activities of respiratory care faculty members in southern academic health centers via a mailed survey. RESULTS: An analysis of the responses (n = 33, 86.8%) revealed: (1) The respondents' principal scholarly activity was the reporting of research findings in refereed journals, with a productivity index (number of articles/years on faculty) of 0.25, or one published article for every 4 years of employment in higher education, which was significantly less than that of other allied health faculty (productivity index 0.69, P less than 0.05). (2) Less than a majority of respondents had presented a paper at a professional meeting during the 3 years preceding the survey. (3) Only a small percentage of respondents had been involved in research. (4) Promotion opportunities and academic preparation are the primary factors that encourage scholarly pursuits, and heavy teaching responsibility is the primary discouraging factor. (5) Scholarly activity is perceived as an important consideration in academic promotion decisions. CONCLUSION: Respiratory care program faculty and administration should take steps to increase the scholarly production of faculty members.

Partial characterization of phospholipase C activity in normal, psoriatic uninvolved, and lesional epidermis.

Arachidonic acid (AA), the precursor of prostaglandins and leukotrienes, can be directly liberated from membrane phospholipids by phospholipase A2 or indirectly by phospholipase C. One or both of these enzymes may be responsible for the increased content of AA found in psoriatic lesional epidermis. Keratome biopsies were obtained from normal and psoriatic individuals. After homogenization and sonication, a 10,000 g supernatant was used as the enzyme source. The activities of both phospholipase A2 and C were assayed in each sample using phosphatidylcholine and phosphatidylinositol, respectively, as substrates. Phospholipase A2 activity was found to be significantly higher than normal in both uninvolved and lesional psoriatic epidermis. In contrast, phospholipase C activity was significantly higher than normal in only the psoriatic plaque on the basis of wet weight (p less than 0.001), protein (p = 0.01), and DNA (p = 0.004) content. Phospholipase C activity in pmol diacylglycerol formed/min/microgram DNA was: normal 4.96 +/- 0.80, n = 13; uninvolved 7.29 +/- 1.06, n = 18; plaque 14.44 +/- 2.50, n = 18. Analysis (pH profile, calcium requirement, substrate specificity, and saturation kinetics) of pooled epidermal extracts showed no inherent differences in phospholipase C from normal and psoriatic epidermis, suggesting either a higher concentration or the presence of an activated form of the enzyme in psoriatic plaque. Since phospholipase C activity, in contrast to phospholipase A2 activity, is elevated only in lesional epidermis, it is possible that this enzyme contributes to AA accumulation observed in this tissue.