Cytidine 5'-Diphosphocholine Corrects Alveolar Type II Cell Mitochondrial Dysfunction in Influenza-infected Mice.

Development of acute respiratory distress syndrome (ARDS) in influenza A virus (IAV)-infected mice is associated with inhibition of ATII (alveolar type II) epithelial cell de novo phosphatidylcholine synthesis, and administration of the phosphatidylcholine precursor cytidine 5'-diphosphocholine (CDP-choline) attenuates IAV-induced acute respiratory distress syndrome in mice. We hypothesized inhibition of phosphatidylcholine synthesis would also impact the function of ATII cell mitochondria. To test this hypothesis, adult C57BL/6 mice of both sexes were inoculated intranasally with 10,000 pfu/mouse influenza A/WSN/33 (H1N1). Control mice were mock-infected with virus diluent. Mice were treated with saline vehicle or CDP-choline (100 µg/mouse i.p.) once daily from 1 to 5 days postinoculation (dpi). ATII cells were isolated by a standard lung digestion protocol at 6 dpi for analysis of mitochondrial function. IAV infection increased uptake of the glucose analog fludeoxyglucose F 18 by the lungs and caused a switch from oxidative phosphorylation to aerobic glycolysis as a primary means of ATII cell ATP synthesis by 6 dpi. Infection also induced ATII cell mitochondrial depolarization and shrinkage, upregulation of PGC-1α, decreased cardiolipin content, and reduced expression of mitofusin 1, OPA1, DRP1, complexes I and IV of the electron transport chain, and enzymes involved in cardiolipin synthesis. Daily CDP-choline treatment prevented the declines in oxidative phosphorylation, mitochondrial membrane potential, and cardiolipin synthesis resulting from IAV infection but did not fully reverse the glycolytic shift. CDP-choline also did not prevent the alterations in mitochondrial protein expression resulting from infection. Taken together, our data show ATII cell mitochondrial dysfunction after IAV infection results from impaired de novo phospholipid synthesis, but the glycolytic shift does not.

Development of a nanoparticle-based tendon-targeting drug delivery system to pharmacologically modulate tendon healing.

Satisfactory healing following acute tendon injury is marred by fibrosis. Despite the high frequency of tendon injuries and poor outcomes, there are no pharmacological therapies in use to enhance the healing process. Moreover, systemic treatments demonstrate poor tendon homing, limiting the beneficial effects of potential tendon therapeutics. To address this unmet need, we leveraged our existing tendon healing spatial transcriptomics dataset and identified an area enriched for expression of Acp5 (TRAP) and subsequently demonstrated robust TRAP activity in the healing tendon. This unexpected finding allowed us to refine and apply our existing TRAP binding peptide (TBP) functionalized nanoparticle (NP) drug delivery system (DDS) to facilitate improved delivery of systemic treatments to the healing tendon. To demonstrate the translational potential of this DDS, we delivered niclosamide (NEN), an S100a4 inhibitor. While systemic delivery of free NEN did not alter healing, TBP-NPNEN enhanced both functional and mechanical recovery, demonstrating the translational potential of this approach to enhance the tendon healing process.

CD30+ lymphomatoid drug reactions.

We report 5 cases of cutaneous CD30+ lymphomatoid drug reactions that occurred shortly after the onset of drug exposure and resolved promptly upon withdrawal of the offending agents. The cases showed protean dermatologic manifestations ranging from diffuse erythema with desquamation to macules, papules, and annular plaques. The suspect drugs were amlodipine (a calcium channel blocker) for 2 cases, sertraline (a selective serotonin reuptake inhibitor) for 1 case, gabapentin for 1 case, and levofloxacin (a fluoroquinolone) versus cefepime (a fourth generation cephalosporin), and metoprolol (a beta blocker), in the fifth case. The histopathologic findings included varying combinations of spongiotic dermatitis, lichenoid infiltrates, and interface dermatitis with a dermal infiltrate of large atypical lymphocytes. Three of the 5 cases contained as much as 30% CD30+ staining of all lymphocytes, whereas the remaining 2 showed 5%-15% positivity. Three patients had a history of allergy or immune dysregulation. Increased knowledge of CD30 positivity in lymphomatoid drug reactions may be relevant in an era of targeted drug therapies. Recognition of these findings may help clinicians to tailor appropriate clinical evaluation and treatment including a review of medications and the removal of possible offending agents.

Development of a Nanoparticle-Based Tendon-Targeting Drug Delivery System to Pharmacologically Modulate Tendon Healing.

Tendon regeneration following acute injury is marred by a fibrotic healing response that prevents complete functional recovery. Despite the high frequency of tendon injuries and the poor outcomes, including functional deficits and elevated risk of re-injury, there are currently no pharmacological therapies in clinical use to enhance the healing process. Several promising pharmacotherapies have been identified; however, systemic treatments lack tendon specificity, resulting in poor tendon biodistribution and perhaps explaining the largely limited beneficial effects of these treatments on the tendon healing process. To address this major unmet need, we leveraged our existing spatial transcriptomics dataset of the tendon healing process to identify an area of the healing tendon that is enriched for expression of Acp5. Acp5 encodes tartrate-resistant acid phosphatase (TRAP), and we demonstrate robust TRAP activity in the healing tendon. This unexpected finding allowed us to refine and apply our existing TRAP binding peptide (TBP) functionalized nanoparticle (NP) drug delivery system (DDS) to facilitate improved delivery of systemic treatments to the healing tendon. To demonstrate the translational potential of this drug delivery system, we delivered the S100a4 inhibitor, Niclosamide to the healing tendon. We have previously shown that genetic knockdown of S100a4 enhances tendon healing. While systemic delivery of Niclosamide did not affect the healing process, relative to controls, TBP-NP delivery of Niclosamide enhanced both functional and mechanical outcome measures. Collectively, these data identify a novel tendon-targeting drug delivery system and demonstrate the translational potential of this approach to enhance the tendon healing process.

Moisturizers: reality and the skin benefits.

The function of the skin as a barrier protects underlying tissues from infection, desiccation, chemicals, and mechanical stress. Disruption of this function results in increased transepidermal water loss or TEWL and is associated with conditions like atopic dermatitis and other chronic skin diseases. Moisturizers have been shown to improve these conditions through restoration of the integrity of the stratum corneum, acting as a barrier to water loss and replacement of skin lipids and other compounds. Also, moisturizers are commonly used to reduce fine lines and make skin appear smooth and soft. While many products make extensive claims of skin rejuvenation, many of the beneficial effects of these products are actually due to the moisturizers they contain: ingredients like glycerin, petrolatum, and dimethicone. Some newer formulations like prescription-device moisturizers, which received 510 K approval on the basis of reducing TEWL, are significantly more expensive than traditional moisturizers and recent literature does not indicate that they are more effective than their over-the-counter counterparts.

Over-the-counter topical skincare products: a review of the literature.

Topical "anti-aging" products, with their seemingly limitless list of ingredients, make extensive claims to reduce wrinkles, fine lines, and sun damage, among others. Sales in the United States alone for cosmeceutical products are expected to increase by 7.4% per year to $8.2 billion by 2012. However, in this enormous industry, there has been a significant lack of rigorous controlled trials of efficacy. It is difficult for both dermatologists and consumers to make informed decisions in a market that is yet to be clearly defined and regulated. We elucidate the scientific basis for, as well as the literature behind, common active ingredients found in products intended to reverse photoaging, discuss some interesting new activities, and provide a review of several comprehensive studies on over-the-counter (OTC) products.

Lindane toxicity: a comprehensive review of the medical literature.

Lindane is an organochloride pesticide, first introduced as a scabicide for human use in the 1950s. Because of its low cost and efficacy, it quickly became a first-line treatment for scabies and head lice, but not long after its use became widespread, its safety was called into question, primarily regarding its neurotoxic effects. We intend to show through review of the literature and databases that lindane has been associated with numerous severe and fatal adverse reactions. This review will summarize 67 cases of adverse reactions and deaths associated with the medical use of lindane. Many of the most serious events and fatalities occurred in pediatric and geriatric populations. We concede that toxicity frequently arises from misuse, but labeled usage of lindane caused 43% of the serious adverse reactions of this review. Moreover, the numerous instances of lindane misuse elucidate the importance of clear labeling and instructions, especially with a product with a narrow therapeutic index. Therefore, this report attempts to elucidate the dangers of lindane as a medical treatment for scabies and head lice by providing a comprehensive review of all documented adverse drug reactions since its introduction.

Developmental cardiac hypertrophy in a mouse model of prolidase deficiency.

BACKGROUND: Hypertrophic cardiomyopathy, characterized by thickened ventricular walls and reduced ventricular chamber volume, is a common cause of sudden cardiac death in young people. Most inherited forms result from mutations in genes encoding sarcomeric proteins. METHODS: Histologic analysis identified embryonic cardiac hypertrophy in dark-like mutant mice. BrdU analysis was performed to measure proliferation and cardiomyocytes were isolated to measure cell size. The dark-like mutation was identified by positional cloning. RESULTS: The dark-like mutation causes cardiomyocyte hypertrophy due to loss-of-function of peptidase d (Pepd), which encodes prolidase, a cytosolic enzyme that recycles proline for collagen re-synthesis. Prolidase deficiency is a rare autosomal recessive disease in humans with a broad phenotypic spectrum not reported to include heart defects, but a conserved role for prolidase in heart development was confirmed by morpholino knockdown in zebrafish. We tested the hypothesis that loss of prolidase function disrupts collagen-mediated integrin signaling and determined that the levels of several key integrin transducers were reduced in the hearts of dark-like mutant embryos. CONCLUSIONS: This work identifies dark-like mice as a model of prolidase deficiency that will be valuable for studying the role of proline metabolism in normal physiology and disease processes, and suggests that integrin signaling may regulate the onset of hypertrophic cardiac growth.

Evaluation of the Sterility of Reynolds Wrap Aluminum Foil for Use During Rodent Surgery.

In biomedical research, surgeons are often responsible for simultaneously conducting rodent surgical procedures, monitoring anesthesia, and adjusting nonsterile equipment. Maintaining appropriate aseptic technique can be challenging when working under these conditions. Applying a sterile barrier material such as aluminum foil to nonsterile surfaces in these circumstances offers an innovative, inexpensive option to improve asepsis. The purpose of this study was to validate the sterility of foodgrade aluminum foil for use as a sterile barrier on nonsterile equipment during rodent surgery. In this investigation, 10 boxes of aluminum foil were assessed for sterility by using ATP swabs and replicate organism detection and counting (RODAC) plates at 0, 14, and 28 d and 6 mo. At 6 mo, foil was applied to surgical equipment, and sterility was assessed by using ATP swabs and RODAC plates. Results revealed no ATP-positive results at any time point. During assessment of samples obtained directly from boxes, RODAC plates yielded minimal bacterial growth (1 cfu per plate) in 2 of the 10 boxes at initial testing and in 1 box at the day 0, day 14, and 6 mo time points. No growth was observed at day 28 (tested directly from the box) or at 6-mo apparatus testing. Our data revealed minimal bacterial growth on tested samples and support the use of Reynolds Wrap aluminum foil as a sterile barrier on nonsterile surfaces during aseptic rodent surgery.

Metabolic shifts modulate lung injury caused by infection with H1N1 influenza A virus.

Influenza A virus (IAV) infection alters lung epithelial cell metabolism in vitro by promoting a glycolytic shift. We hypothesized that this shift benefits the virus rather than the host and that inhibition of glycolysis would improve infection outcomes. A/WSN/33 IAV-inoculated C57BL/6 mice were treated daily from 1 day post-inoculation (d.p.i.) with 2-deoxy-d-glucose (2-DG) to inhibit glycolysis and with the pyruvate dehydrogenase kinase (PDK) inhibitor dichloroacetate (DCA) to promote flux through the TCA cycle. To block OXPHOS, mice were treated every other day from 1 d.p.i. with the Complex I inhibitor rotenone (ROT). 2-DG significantly decreased nocturnal activity, reduced respiratory exchange ratios, worsened hypoxemia, exacerbated lung dysfunction, and increased humoral inflammation at 6 d.p.i. DCA and ROT treatment normalized oxygenation and airway resistance and attenuated IAV-induced pulmonary edema, histopathology, and nitrotyrosine formation. None of the treatments altered viral replication. These data suggest that a shift to glycolysis is host-protective in influenza.

Repeated Orotracheal Intubation in Mice.

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ventral neck for direct confirmation of cannula placement in the trachea. The relative difficulty or the tissue trauma induced to the subject by such procedures can be an impediment to an investigator's ability to perform longitudinal studies. This article illustrates a technique in which physical manipulation of the mouse following the use of a depilatory to remove hair from the ventral neck permits transcutaneous visualization of the trachea for orotracheal intubation regardless of degree of skin pigmentation. This method is innocuous to the subject and easily achieved with a limited understanding of murine anatomy. This refined approach facilitates repeated intubation, which may be necessary for monitoring progression of disease or instillation of treatments. Using this method may result in a reduction of the number of animals and technical skill required to measure lung function in mouse models of respiratory disease.

Oropharyngeal morphology related to filtration mechanisms in suspension-feeding American shad (Clupeidae).

To assess potential filtration mechanisms, scanning electron microscopy was used in a comprehensive quantification and analysis of the morphology and surface ultrastructure for all five branchial arches in the ram suspension-feeding fish, American shad (Alosa sapidissima, Clupeidae). The orientation of the branchial arches and the location of mucus cells on the gill rakers were more consistent with mechanisms of crossflow filtration and cross-step filtration rather than conventional dead-end sieving. The long, thin gill rakers could lead to a large area for the exit of water from the oropharyngeal cavity during suspension feeding (high fluid exit ratio). The substantial elongation of gill rakers along the dorsal-ventral axis formed d-type ribs with a groove aspect ratio of 0.5 and a Reynolds number of approximately 500, consistent with the potential operation of cross-step filtration. Mucus cell abundance differed significantly along the length of the raker and the height of the raker. The mucus cell abundance data and the observed sloughing of denticles along the gill raker margins closest to the interior of the oropharyngeal cavity suggest that gill raker growth may occur primarily at the raker tips, the denticle bases, and the internal raker margins along the length of the raker. These findings will be applied in ongoing experiments with 3D-printed physical models of fish oral cavities in flow tanks, and in future ecological studies on the diet and nutrition of suspension-feeding fishes.

Barriers to Enacting Childhood Sun Safety Behavior: Findings from Focus Group Interviews Among Hispanic Parents in Miami.

Hispanics are generally diagnosed at more advanced stages of melanoma than non-Hispanic Whites, leading to lower survival rates. As skin cancer incidence is attributable to lifetime exposure to ultraviolet light, encouraging the performance of sun safety behaviors in childhood is an important strategy to address this divide. Problematically, we know little about the barriers to sun safety among Hispanic youth, especially among the Hispanics living in South Florida. To address this gap, we conducted focus groups among parents of Hispanic children aged 4-10 to understand the unique barriers to sun protection among this audience. Results revealed four categories of barriers: child-based barriers, external barriers, parental enactment barriers, and parental proper adherence barriers. These results are discussed in terms of their implications for future intervention research among this audience.

Tissue engineering and biomaterials in regenerative medicine.

The field of regenerative medicine offers the potential to significantly impact a wide spectrum of healthcare issues, from diabetes to cardiovascular disease. In particular, the design of tailored biomaterials, which possess properties desired for their particular application, and the development of superior implant environments, which seek to meet the nutritional needs of the tissue, have yielded promising tissue engineering prototypes. In this commentary, we examine the novel approaches researchers have made in customized biomaterials and promoting angiogenesis that have led to significant advancements in recent years.

Pathway-based analysis of breast cancer.

INTRODUCTION: Although HER2 and ER pathways are predominant pathways altered in breast cancer, it is now well accepted that many other signaling pathways are also involved in the pathogenesis of breast cancer. The understanding of these additional pathways may assist in identifying new therapeutic approaches for breast cancer. METHODS: 13 invasive ductal carcinoma tissues and 5 benign breast tissues were analyzed for the mRNA expression level of 1243 cancer pathway-related genes using SmartChip (WaferGen, CA), a real-time PCR-base method. In addition, the levels of 131 cancer pathway-related proteins and phosphoproteins in 33 paired breast cancers were measured using our innovative Protein Pathway Array. RESULTS: Out of 1,243 mRNAs, 68.7% (854) were detected in breast cancer and 395 mRNAs were statistically significant (fold change >2) between benign and cancer tissues. Of these mRNAs, 105 only expressed in breast cancer tissues and 33 mRNAs only expressed in normal breast tissues. Out of 131 proteins and phosphoproteins, 68% (89) were detected in cancer tissues and 57 proteins were significantly differentiated between tumor and normal tissues. Interestingly, only 3 genes (CDK6, Vimentin and SLUG) showed decreases in both protein and mRNA. Six proteins (BCL6, CCNE1, PCNA, PDK1, SRC and XIAP) were differentially expressed between tumor and normal tissues but no differences were observed at mRNA levels. Analyses of mRNA and protein data using Ingenuity Pathway Analysis showed more than 15 pathways were altered in breast cancer and 6 of which were shared between mRNAs and proteins, including p53, IL17, HGF, NGF, PTEN and PI3K/AKT pathways. CONCLUSIONS: There is a broad dysregulation of various pathways in breast cancer both at protein levels and mRNA levels. It is important to note that mRNA expression does not correlate with protein level, suggesting different regulation mechanisms between proteins and mRNAs.

Melanoma arising in a tattoo: case report and review of the literature.

Various benign and malignant lesions have been described in relation to tattoos including melanoma. Few cases of malignant melanoma (MM) arising in tattoos have been reported in the literature. We report a 79-year-old man with an MM that arose in a tattoo he had for 60 years on the inferior aspect of the left arm. This case underscores the need for careful examination of tattoos to insure that dysplastic or malignant pigmented lesions are not overlooked. We also discuss the possibility of a pathogenic relationship between MM and tattoos.