Misidentification of Saprochaete clavata as Magnusiomyces capitatus in Clinical Isolates: Utility of Internal Transcribed Spacer Sequencing and Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry and Importance of Reliable Databases

Saprochaete clavata and Magnusiomyces capitatus are human pathogens that are frequently mistaken for each other due to their similar phenotypes and erroneous or limited databases. Based on internal transcribed spacer (ITS) sequences, we propose species-specific carbon assimilation patterns and matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) fingerprints that enable the identification of S. clavata, M. capitatus, and Galactomyces candidus to the species level.     

The role of the gallbladder in the pathogenesis of cholesterol gallstones.

During the past century, a variety of explanations have been proposed to explain the pathogenesis of cholesterol gallstones. Early attempts to account for the phenomenon of cholelithiasis focused on events in the gallbladder and stressed mucosal inflammatory changes, gallbladder stasis, stratification of bile, and absorption of bile salts from a damaged mucosa. The advent of the concept of "lithogenic bile" redirected attention to the liver and led to the proposal that an enzyme-mediated genetic and/or metabolic defect is the initiator of cholesterol cholelithiasis. While recognizing that the pathogenesis of gallstones is probably multifactorial, alterations in gallbladder and biliary ductal motor function constitute a plausible, but as yet unexplored, mechanism for alterations in enterohepatic circulation dynamics and subsequent cholesterol cholelithiasis. Gallbladder motor function is a complex phenomenon influenced by dynamic compliance, autonomic pharmacology, hormonal responses, and sphincter dynamics. Attempts to describe these aspects of biliary physiology may characterize the next phase in our understanding of the pathogenesis of cholesterol cholelithiasis.     

Inducing Humoral and Cellular Responses to Multiple Sporozoite and Liver-Stage Malaria Antigens Using Exogenous Plasmid DNA

A vaccine candidate that elicits humoral and cellular responses to multiple sporozoite and liver-stage antigens may be able to confer protection against Plasmodium falciparum malaria; however, a technology for formulating and delivering such a vaccine has remained elusive. Here, we report the preclinical assessment of an optimized DNA vaccine approach that targets four P. falciparum antigens: circumsporozoite protein (CSP), liver stage antigen 1 (LSA1), thrombospondin-related anonymous protein (TRAP), and cell-traversal protein for ookinetes and sporozoites (CelTOS). Synthetic DNA sequences were designed for each antigen with modifications to improve expression and were delivered using in vivo electroporation (EP). Immunogenicity was evaluated in mice and nonhuman primates (NHPs) and assessed by enzyme-linked immunosorbent assay (ELISA), gamma interferon (IFN-γ) enzyme-linked immunosorbent spot (ELISpot) assay, and flow cytometry. In mice, DNA with EP delivery induced antigen-specific IFN-γ production, as measured by ELISpot assay and IgG seroconversion against all antigens. Sustained production of IFN-γ, interleukin-2, and tumor necrosis factor alpha was elicited in both the CD4⁺ and CD8⁺ T cell compartments. Furthermore, hepatic CD8⁺ lymphocytes produced LSA1-specific IFN-γ. The immune responses conferred to mice by this approach translated to the NHP model, which showed cellular responses by ELISpot assay and intracellular cytokine staining. Notably, antigen-specific CD8⁺ granzyme B⁺ T cells were observed in NHPs. Collectively, the data demonstrate that delivery of gene sequences by DNA/EP encoding malaria parasite antigens is immunogenic in animal models and can harness both the humoral and cellular arms of the immune system.     

Vaccinia‐based influenza vaccine overcomes previously induced immunodominance hierarchy for heterosubtypic protection

Growing concerns about unpredictable influenza pandemics require a broadly protective vaccine against diverse influenza strains. One of the promising approaches was a T cell‐based vaccine, but the narrow breadth of T‐cell immunity due to the immunodominance hierarchy established by previous influenza infection and efficacy against only mild challenge condition are important hurdles to overcome. To model T‐cell immunodominance hierarchy in humans in an experimental setting, influenza‐primed C57BL/6 mice were chosen and boosted with a mixture of vaccinia recombinants, individually expressing consensus sequences from avian, swine, and human isolates of influenza internal proteins. As determined by IFN‐γ ELISPOT and polyfunctional cytokine secretion, the vaccinia recombinants of influenza expanded the breadth of T‐cell responses to include subdominant and even minor epitopes. Vaccine groups were successfully protected against 100 LD₅₀ challenges with PR/8/34 and highly pathogenic avian influenza H5N1, which contained the identical dominant NP₃₆₆ epitope. Interestingly, in challenge with pandemic A/Cal/04/2009 containing mutations in the dominant epitope, only the group vaccinated with rVV‐NP + PA showed improved protection. Taken together, a vaccinia‐based influenza vaccine expressing conserved internal proteins improved the breadth of influenza‐specific T‐cell immunity and provided heterosubtypic protection against immunologically close as well as distant influenza strains.     

Vancomycin-Variable Enterococcus faecium: In Vivo Emergence of Vancomycin Resistance in a Vancomycin-Susceptible Isolate

We report the emergence of vancomycin resistance in a patient colonized with a vanA-containing, vanRS-negative isolate of Enterococcus faecium which was initially vancomycin susceptible. This is a previously undescribed mechanism of drug resistance with diagnostic and therapeutic implications.     

The Associations of Body Composition and Fat Distribution With Bone Mineral Density in Elderly Italian Men and Women

This study aimed to investigate the associations of body composition and fat distribution with bone mineral density (BMD) in elderly Italian subjects. In 866 women (age 64.2 ± 6.5 yr) and 168 men (age 65.1 ± 6.1 yr), we measured BMD at lumbar spine, at femur, at the total body, and at the right hand. In all subjects, we also measured sex hormones, 25-hydroxyvitamin D, bone markers, and calcium intake. In both men and women, all body composition parameters had significant positive correlations with BMD at all sites after adjusting for age only; after adjusting also for body weight only lean mass (LM) remained positively associated with BMD at all sites except BMD at lumbar spine. In males, LM was associated with BMD at all sites, whereas android fat was associated with BMD at lumbar spine, at femur, and at whole body. In females, fat mass (FM) was positively and age inversely associated with BMD at all sites, whereas gynoid fat and alkaline phosphatase were inversely associated with BMD at lumbar spine and at femur. In conclusion, the role of LM seems more important in males, whereas in women the role of FM prevails with negative associations between gynoid fat and BMD.     

Olfactory responses of medically and economically important mites (Acari: Epidermoptidae and Acaridae) to volatile chemicals

Dermatophagoides farinae Hughes (Acari: Epidermoptidae), the American house dust mite, and Tyrophagus putrescentiae (Schrank) (Acari: Acaridae), the mold mite, are medically and economically important but controlling them has proved difficult, and recolonization is commonplace. Their behavioral responses to different sources of volatile chemicals are still not fully elucidated. For the first time, the Y-tube olfactometer, which is an enclosed bioassay to resolve responses to test and control volatiles, has been successfully used with these mites. Mites were tested individually, and both T. putrescentiae and D. farinae responded to food volatiles. Y-tube olfactometers may be used to test for potential semiochemicals, thereby increasing knowledge of our behavior of astigmatic mites.     

Prospective analysis of adverse events in surgical treatment of degenerative spondylolisthesis

Background contextSurgical adverse event (AE) monitoring is imprecise, of uncertain validity, and tends toward underreporting. Reports focus on specific procedures rather than outcomes in the context of presenting diagnosis. Specific intraoperative (intraop) or postoperative (postop) AEs that may be independently associated with degenerative spondylolisthesis (DS) have never been reported.PurposeThe primary purpose was to assess the AE profile of surgically treated patients with L4–L5 DS. The secondary goal was to identify potential risk factors that correlate with those AEs.Study design/settingProspective cohort and academic quaternary spine center.Patient sampleNinety-two patients with L4–L5 DS were treated surgically, discharged from Vancouver General Hospital between January 1, 2009 and December 31, 2010.Outcome measuresIncidence rates and odds ratios.MethodsProspective AE data were analyzed using univariate analyses, forward selection regression models, and Spearman correlation coefficients. Results were compared with outcomes reported in the Spine Patient Outcomes Research Trial.ResultsNo AEs were seen in 57.6% of patients, one AE in 17.4%, and two or more AEs in 17.4%. Dural tears (6.5%) and intraop bone-implant interface failure requiring revision (3.3%) were the most common intraop AEs. Postoperatively, the most frequent AEs were urinary tract infection (10.9%), delirium (5.4%), neuropathic pain (4.4%), deep wound infection (3.3%), and superficial wound infection (3.3%). The odds of an intraop AE increased by 9% (95% confidence interval [CI] 1–18) per year of age at admission. Adjusted Charlson comorbidity index (CCI) did not correlate with number of AEs experienced. The odds of postop delirium correlated with CCI (odds ratio [OR] 3.39, 95% CI 1.12–10.24) and dural tear (OR 35.84, 95% CI 1.72–747.45). Length of stay was statistically significant and was influenced by two or more AEs, CCI, postop loss of correction, cerebrospinal fluid leak, deep wound infection, noninfected wound drainage, and gender.ConclusionsRisk of intraop AEs, but not postop AEs, increased with increasing age. Having multiple comorbidities does not predispose to more AEs. Infections predominate among the postop AEs. Patients at increased risk of delirium or of having an increased length of hospital stay may more easily be predicted. Studies specifically designed to prospectively assess AEs have the potential to more accurately identify postop AE rates.     

Myriocin induces apoptotic lung cancer cell death via activation of DR4 pathway

It has been known that myriocin inhibits melanoma growth. However, the effects and action mechanisms of myriocin on lung cancer cell growth have not been reported. In this study, we examined whether myriocin isolated from Mycelia sterilia inhibits cell growth of lung cancer cells (A549 and NCI-H460) as well as possible signaling pathways involved in cell growth inhibition. Different concentrations of myriocin inhibited the growth of lung cancer cells through the induction of apoptotic cell death. Consistent with cancer cell growth inhibition, myriocin induced the expression of death receptors (DRs) as well as p-JNK and p-p38 in both cell lines. Moreover, the combination of myriocin with DR4 ligand TRAIL, and other well known anti-tumor drugs (docetaxel and cisplatin) synergistically inhibited cancer cell growth, and induced DR4 expression. These results showed that myriocin inhibits lung cancer cells growth through apoptosis via the activation of DR4 pathways, and enhanced anti-cancer effects with well known drugs. Thus, our study indicates that myriocin could be effective for lung cancer cells as an anti-cancer drug and/or a conjunction agent with well known anti-cancers.