Hydroxyl functionalized multi-walled carbon nanotubes modulate immune responses without increasing 2009 pandemic influenza A/H1N1 virus titers in infected mice.

Growing use of carbon nanotubes (CNTs) have garnered concerns regarding their association with adverse health effects. Few studies have probed how CNTs affect a host's susceptibility to pathogens, particularly respiratory viruses. We reported that exposure of lung cells and mice to pristine single-walled CNTs (SWCNTs) leads to significantly increased influenza virus H1N1 strain A/Mexico/4108/2009 (IAV) titers in concert with repressed antiviral immune responses. In the present study, we investigated if hydroxylated multi-walled CNTs (MWCNTs), would result in similar outcomes. C57BL/6 mice were exposed to 20 µg MWCNTs on day 0 and IAV on day 3 and samples were collected on day 7. We investigated pathological changes, viral titers, immune-related gene expression in lung tissue, and quantified differential cell counts and cytokine and chemokine levels in bronchoalveolar lavage fluid. MWCNTs alone caused mild inflammation with no apparent changes in immune markers whereas IAV alone presented typical infection-associated inflammation, pathology, and titers. The co-exposure (MWCNTs + IAV) did not alter titers or immune cell profiles compared to the IAV only but increased concentrations of IL-1ß, TNFα, GM-CSF, KC, MIPs, and RANTES and inhibited mRNA expression of Tlr3, Rig-i, Mda5, and Ifit2. Our findings suggest MWCNTs modulate immune responses to IAV with no effect on the viral titer and modest pulmonary injury, a result different from those reported for SWCNT exposures. This is the first study to show that MWCNTs modify cytokine and chemokine responses that control aspects of host defenses which may play a greater role in mitigating IAV infections.

Mycotic Rhinitis and Sinusitis in Florida Horses.

Rhinitis and sinusitis caused by fungal pathogens were studied in biopsy samples submitted from 52 horses distributed throughout subtropical and tropical regions of Florida. Methods included routine histopathology as well as polymerase chain reaction (PCR) with panfungal/panoomycete primers and DNA sequencing on extracted DNA (DNA barcoding). Granulomatous, pyogranulomatous, and fibrinopurulent lesions in nasal and sinus mucosa were associated with signs of upper airway obstruction and noise as well as nasal discharge. Morphologic and histochemical assessment of cases identified 31 cases of zygomycosis/pythiosis plus 1 mixed infection case, 16 cases of phaeohyphomycosis with 2 additional mixed infection cases, and 3 cases caused by other fungi. Morphologic evidence of Aspergillus sp. infection as a superficial copathogen was found in 2 of the mixed fungal infection cases. PCR and DNA sequencing facilitated identification of fungal pathogens in 11 of 52 cases (21%). No evidence of oomycete infection was found. Histomorphologic features of previously unrecognized forms of equine rhinitis/sinusitis were described, including those caused by Flavodon flavus, Curvularia lunata, Exserohilum rostrata, Alternaria alternata, Alternaria sp., Cladophialophora bantiana, Fusarium solani, and Toxicocladosporium irritans. PCR and DNA sequencing using panfungal and oomycete primers with DNA from formalin-fixed and paraffin-embedded specimens successfully identified the pathogen in phaeohyphomycosis (7/18 cases, 39%), zygomycosis/pythiosis (1/32 cases, 3%), and other nonpigmented fungal infections (3/3 cases, 100%). Zygomycosis and phaeohyphomycosis were the most common forms of fungal rhinitis found in Florida horses.

α-Galactosylceramide protects swine against influenza infection when administered as a vaccine adjuvant.

Natural killer T (NKT) -cells activated with the glycolipid ligand α-galactosylceramide (α-GalCer) stimulate a wide array of immune responses with many promising immunotherapeutic applications, including the enhancement of vaccines against infectious diseases and cancer. In the current study, we evaluated whether α-GalCer generates protective immunity against a swine influenza (SI) virus infection when applied as an intramuscular vaccine adjuvant. Immunization of newly weaned piglets with UV-killed pandemic H1N1 A/California/04/2009 (kCA04) SI virus and α-GalCer induced high titers of anti-hemagglutinin antibodies and generated virus-specific T cells that localized in intrapulmonary airways and in alveolar walls. Vaccination with α-GalCer resulted in a systemic increase in NKT-cell concentrations, including in the respiratory tract, which was associated with complete inhibition of viral replication in the upper and lower respiratory tract and much reduced viral shedding. These results indicate that NKT-cell agonists could be used to improve swine vaccine formulations in order to reduce the clinical signs of SI infection and limit the spread of influenza viruses amongst commercial pigs.

A comparison of two methods of endoscopic dilation of acute subglottic stenosis using a ferret model.

OBJECTIVES/HYPOTHESIS: Balloon dilation is accepted as a first line treatment of acute subglottic stenosis, but its effects on the subglottic tissue remain largely unknown. We aimed to develop an animal model of acute subglottic stenosis using endoscopic techniques. Once developed, this model was used to compare the immediate effects of balloon dilation and endotracheal tube dilation on subglottic tissue. STUDY DESIGN: Prospective randomized animal study. METHODS: Acute subglottic injury was induced in 10 ferrets by endoscopic cauterization with silver nitrate. After 48-72 hours of observation, eight animals were randomized to undergo subglottic dilation with either a 5-mm balloon or endotracheal tubes of increasing diameter. These eight ferrets were euthanized within 10 minutes after dilation. The other two ferrets served as controls and were euthanized following observation only. The larynx from each ferret was harvested, and the subglottis was examined histologically by a pathologist blinded to the treatment arms. RESULTS: Acute subglottic stenosis was induced in all 10 ferrets using the endoscopic technique. Both balloon and endotracheal tube dilation resulted in comparable improvement in the subglottic airway diameter. A decreased thickness of submucosa/lamina propria was seen in the balloon dilation group. CONCLUSIONS: Acute subglottic stenosis can be reliably induced in ferrets using endoscopic techniques. Multiple dilation methods can be used to relieve acute obstruction. Balloon dilators seem to improve airway patency, in part, by decreasing the thickness of the submucosa and lamina propria. Further research is needed to determine how this impacts later stages of wound healing and final outcomes.

Comparative evaluation of differential laser-induced perturbation spectroscopy as a technique to discriminate emerging skin pathology.

Fluorescence spectroscopy has been widely investigated as a technique for identifying pathological tissue; however, unrelated subject-to-subject variations in spectra complicate data analysis and interpretation. We describe and evaluate a new biosensing technique, differential laser-induced perturbation spectroscopy (DLIPS), based on deep ultraviolet (UV) photochemical perturbation in combination with difference spectroscopy. This technique combines sequential fluorescence probing (pre- and post-perturbation) with sub-ablative UV perturbation and difference spectroscopy to provide a new spectral dimension, facilitating two improvements over fluorescence spectroscopy. First, the differential technique eliminates significant variations in absolute fluorescence response within subject populations. Second, UV perturbations alter the extracellular matrix (ECM), directly coupling the DLIPS response to the biological structure. Improved biosensing with DLIPS is demonstrated in vivo in a murine model of chemically induced skin lesion development. Component loading analysis of the data indicates that the DLIPS technique couples to structural proteins in the ECM. Analysis of variance shows that DLIPS has a significant response to emerging pathology as opposed to other population differences. An optimal likelihood ratio classifier for the DLIPS dataset shows that this technique holds promise for improved diagnosis of epithelial pathology. Results further indicate that DLIPS may improve diagnosis of tissue by augmenting fluorescence spectra (i.e. orthogonal sensing).

Experimental infection of neonatal foals with Rhodococcus equi triggers adult-like gamma interferon induction.

Rhodococcus equi is a facultative intracellular pathogen that causes pneumonia in young foals but does not induce disease in immunocompetent adult horses. Clearance of R. equi depends mainly on gamma interferon (IFN-gamma) production by T lymphocytes, whereas the predominance of interleukin 4 (IL-4) is detrimental. Young foals, like neonates of many other species, are generally deficient in the ability to produce IFN-gamma. The objective of this study was to compare the cytokine profiles, as well as cell-mediated and antibody responses, of young foals to those of adult horses following intrabronchial challenge with R. equi. The lymphoproliferative responses of bronchial lymph node (BLN) cells to concanavalin A were significantly higher in foals than in adult horses. In contrast, adult horses had significantly higher lymphoproliferative responses to R. equi antigens than did foals. Infected foals had significantly lower IL-4 mRNA expression but significantly higher IFN-gamma expression and IFN-gamma/IL-4 ratio in R. equi-stimulated BLN lymphocytes than did infected adults. Infection with R. equi in foals resulted in a significant increase in the percentage of T lymphocytes and CD4(+) T lymphocytes in bronchoalveolar lavage fluid in association with a significant decrease in the percentage of these cell populations in BLNs. Infection of foals also resulted in a marked increase in serum immunoglobulin Ga (IgGa) and IgGb levels, resulting in concentrations in serum that were significantly higher than those of adult horses. This study demonstrates that the immune response to R. equi in foals is not biased toward IL-4 and is characterized by the predominant induction of IFN-gamma.

Simultaneous congenital and acquired extrahepatic portosystemic shunts in two dogs.

Two dogs with simultaneous congenital and acquired portosystemic shunts are reported. The first dog was an eight-month-old, male Golden Retriever with a history of peritoneal effusion, polyuria/polydipsia, and stunted growth. The dog had a microcytic, hypochromic anemia, a mildly elevated AST, and a moderate to severely elevated preprandial and postprandial serum bile acids. Transcolonic portal scintigraphy confirmed the presence of a portosystemic shunt. An intraoperative mesenteric portogram was performed. Two conjoined congenital extrahepatic portosystemic shunts and multiple acquired extrahepatic portosystemic shunts were identified. The second dog was a five-month-old, mixed breed with two week history of peritoneal effusion. Abdominal ultrasound and transcolonic scintigraphy were used to diagnose a portosystemic shunt. A single extrahepatic portosystemic shunt, portal hypertension, and multiple acquired collateral shunts were identified at surgery. The histologic alterations observed in these dogs were consistent with a portosystemic shunt. In these dogs, the presence of congenital and acquired portosystemic shunts and histopathologic findings are considered to represent a combination of congenital portosystemic shunts and noncirrhotic portal hypertension or portal vein hypoplasia.