Tick bite and Lyme disease-related emergency department encounters in New Hampshire, 2010-2014.

Lyme disease (LD) is a common tick-borne disease in New Hampshire (NH). While LD is a reportable condition and cases are counted for public health surveillance, many more people receive care for tick bites or diagnoses of LD than are reflected in surveillance data. NH's emergency department (ED) data system was queried for tick bite and LD-related encounters. Chief complaint text was queried for words related to LD or tick bites. International Classification of Diseases 9th Revision (ICD-9) codes were queried for the LD diagnosis code (088.81). Emergency department patient data were matched to reportable disease data to determine the proportion of ED patients reported to the health department as a suspected LD case. Data were analysed to calculate frequencies for key demographic and reporting characteristics. From 2010 to 2014, 13,615 tick bite or LD-related ED encounters were identified in NH, with most due to tick bites (76%). Of 3,256 patients with a LD-related ED encounter, 738 (23%) were reported to the health department as a suspected LD case. The geographic distribution of ED patients was similar to reported LD cases; however, the regions of the state that experienced higher rates of ED encounters were different than the regions that observed higher rates of reported LD cases. Seasonal distribution of ED encounters peaked earlier than reported LD cases with a second peak in the fall. While age and sex distribution was similar among ED patients and reported LD cases, the rates for children 5 years and younger and adults 65 years and older were greater for ED encounters. Patients frequently visit the ED to seek care for tick bites and suspected LD. Results of ED data analyses can be used to target education, in particular for ED providers and the public through timely distribution of evidence-based educational materials and training programmes.

Subject-specific, multiscale simulation of electrophysiology: a software pipeline for image-based models and application examples.

Many simulation studies in biomedicine are based on a similar sequence of processing steps, starting from images and running through geometric model generation, assignment of tissue properties, numerical simulation and visualization of the results--a process known as image-based geometric modelling and simulation. We present an overview of software systems for implementing such a sequence both within highly integrated problem-solving environments and in the form of loosely integrated pipelines. Loose integration in this case indicates that individual programs function largely independently but communicate through files of a common format and support simple scripting, so as to automate multiple executions wherever possible. We then describe three specific applications of such pipelines to translational biomedical research in electrophysiology.

Electrographic Response of the Heart to Myocardial Ischemia.

Electrocardiographic (ECG) ST segment shifts are often used as markers for detecting myocardial ischemia. Literature suggests that the progression of ischemia, occurs from the endocardium and spreads towards the epicardium, eventually becoming transmural. Our study with animal models has found the progression of ischemia, characterized by ST elevations to be more complex and heterogeneous in its distribution. We used in situ canine preparations, wherein the animals were subjected to demand ischemia by reducing coronary flow and raising the heart rate through atrial pacing. At reduced flow, increasing the heart rate caused pockets of ST elevations to appear variously distributed in the sub-epicardial, midmyocardial and endocardial regions. Further reduction in coronary flow with simultaneous raising of the heart rate, increased the extent and magnitude of ST elevated regions, that in certain cases became transmural.

Effects of fluoro substitution on 4-bromodiphenyl ether (PBDE 3).

It is our hypothesis that fluoro substitution provides a powerful tool to modulate the desired characteristics and to increase the specificity of studies of structure-activity relationships. 4-Bromodiphenyl ether (PBDE 3) and its five corresponding monofluorinated analogues (F-PBDEs 3) have been synthesized and fully characterized (using (1)H, (13)C and (19)F NMR spectroscopy, and mass spectrometry). The accurate structure from X-ray crystal analysis was compared with iterative calculations using semi-empirical self-consistent field molecular-orbital (SCF-MO) models. The compounds studied were 4-bromodiphenyl ether (PBDE 3), the (13)C(6)-isotopically labeled PBDE 3 ((13)C(6)-PBDE 3) and 2-fluoro-4-bromodiphenyl ether (3-2F), 2'-fluoro-4-bromodiphenyl ether (3-2'F), 3-fluoro-4-bromodiphenyl ether (3-3F), 3'-fluoro-4-bromodiphenyl ether (3-3'F), and 4'-fluoro-4-bromodiphenyl ether (3-4'F). Solid-state intermolecular interactions for PBDE 3 and the F-PBDEs 3 isomers are dominated by weak C-H(F,Br)...pi and C-H...F interactions. The C-F bond lengths varied between 1.347 (2) and 1.362 (2) A, and the C4-Br bond length between 1.880 (3) and 1.904 (2) A. These bond lengths are correlated with electron-density differences, as determined by (13)C shifts, but not with the strength of the C-F couplings. The interior ring angles of ipso-fluoro substitution increased (121.9-124.0 degrees ) as a result of hyperconjugation, a phenomenon also predicted by the calculation models. An attraction between the vicinal fluoro and halo substituents (observed in fluoro substituted chlorobiphenyls) was not observed for the bromo substituted F-PBDEs. The influence of a fluoro substituent on the conformation was only observable in PBDEs with di-ortho substitution. Calculated and observed torsion angles showed a positive correlation with increasing van der Waals radii and/or the degree of substitution for mono- to tetra-fluoro, chloro, bromo and methyl substitutions in the ortho positions of diphenyl ether. These findings utilizing F-tagged analogues presented here may prove fundamental to the interpretation of the biological effects and toxicities of these persistent environmental pollutants.

Influence of fluoro-substitution on the planarity of 4-chlorobiphenyl (PCB 3).

Accurate structure determinations by X-ray crystal analysis and computation using semi-empirical self-consistent field molecular orbital calculations are described and compared for five monofluorinated analogues of 4-chlorobiphenyl, i.e. 2-fluoro-4-chlorobiphenyl, 2'-fluoro-4-chlorobiphenyl, 3-fluoro-4-chlorobiphenyl, 3'-fluoro-4-chlorobiphenyl and 4'-fluoro-4-chlorobiphenyl. Intermolecular interactions for all monofluorinated isomers are dominated by phenyl-phenyl stacking and C-H-phenyl interactions. C-F bond lengths varied between 1.341 and 1.374 A, C-Cl between 1.733 and 1.765 A, and both correlate with electron-density differences as determined by (13)C NMR shifts. The interior ring angles at ipso-fluoro substitution increase up to 122.2-124.2 degrees due to hyperconjugation by 2p-pi-orbital overlapping, a phenomenon that was also reflected in the computer calculation. The angles of C-F and C-Cl relative to the aromatic ring for vicinal fluoro- and chloro substituents show an attraction, not a repulsion, between the adjacent F and Cl substituents. This finding is explained on the basis of electron donor and acceptor properties. The dihedral angles of ortho-substituted biphenyls show that monofluoro substitution results in slightly smaller increases compared with chlorine, while additional ortho-fluorination results in little further change in the dihedral angle. In contrast, ortho-chlorination strongly decreases the co-planarity. This is likely to be due to interior ring-angle distortion and the size of the halogen substituent. Fluoro substitution does indeed affect the planarity of the PCB3 analogues, but these effects are minor compared with chloro substitution. Fluorine tagging offers promise for use in in vitro and in vivo studies. Differences in computational versus measured data emphasize the need to use a variety of methods to ascertain the true nature of the physical properties of a compound.

Evaluation of the Cepheid GeneXpert system for detecting Bacillus anthracis.

AIMS: The Cepheid GeneXpert is a four-site, automated sample preparation and real-time PCR detection system. In this study, the capability of the GeneXpert to isolate and detect nucleic acid from Bacillus anthracis Ames spores was assessed. METHODS AND RESULTS: A four-plex, dried-down bead cartridge containing PCR reagents specific for the pXO1 and pXO2 plasmids as well as sample processing and inhibition controls was evaluated. For B. anthracis Ames spores harbouring pXO1 and pXO2, samples containing 68 CFU per ml (148 spores per ml) were positive in all four replicates. A limited cross-reactivity panel, which included closely related Bacillus species, was also tested to determine the specificity of the pXO1 and pXO2 assays. No cross-reactivity occurred. Further, B. anthracis Sterne spore samples were analysed to compare results when processed using the GeneXpert to those run directly on the Cepheid SmartCycler without sample processing. The GeneXpert detection capability was three logs lower than the SmartCycler indicating the benefit of incorporating a nucleic acid extraction procedure. CONCLUSIONS: This study demonstrates that the GeneXpert is a rapid and reliable system for simultaneously detecting the B. anthracis virulence plasmids pXO1 and pXO2. SIGNIFICANCE AND IMPACT OF THE STUDY: The GeneXpert is the only platform currently available that is capable of both nucleic acid purification and real-time PCR detection enclosed within a single system. Further, all sample manipulations are automated, thus reducing errors associated with manual processing.

Pseudodestruxins A and B: new cyclic depsipeptides from the coprophilous fungus Nigrosabulum globosum.

Pseudodestruxins A (1) and B (2), two new cyclic peptides, have been isolated from cultures of the coprophilous fungus Nigrosabulum globosum. The structure of pseudodestruxin A (1) was elucidated using 2D NMR techniques and confirmed by single-crystal X-ray diffraction analysis. The structure of 2 was assigned by comparing its NMR and FABMS data with those of compound 1. The known compounds ascochlorin and 5-chlorocollectorin B were also isolated from N. globosum. Although 1 and 2 display antibacterial effects, ascochlorin was found to be responsible for the antifungal activity of the crude extract.

Persistence of Mycobacterium tuberculosis in macrophages and mice requires the glyoxylate shunt enzyme isocitrate lyase.

Mycobacterium tuberculosis claims more human lives each year than any other bacterial pathogen. Infection is maintained in spite of acquired immunity and resists eradication by antimicrobials. Despite an urgent need for new therapies targeting persistent bacteria, our knowledge of bacterial metabolism throughout the course of infection remains rudimentary. Here we report that persistence of M. tuberculosis in mice is facilitated by isocitrate lyase (ICL), an enzyme essential for the metabolism of fatty acids. Disruption of the icl gene attenuated bacterial persistence and virulence in immune-competent mice without affecting bacterial growth during the acute phase of infection. A link between the requirement for ICL and the immune status of the host was established by the restored virulence of delta icl bacteria in interferon-gamma knockout mice. This link was apparent at the level of the infected macrophage: Activation of infected macrophages increased expression of ICL, and the delta icl mutant was markedly attenuated for survival in activated but not resting macrophages. These data suggest that the metabolism of M. tuberculosis in vivo is profoundly influenced by the host response to infection, an observation with important implications for the treatment of chronic tuberculosis.

A descriptive system for lower extremity evaluation in children: data for the newborn infant.

An efficient and practical system for examination of the lower extremities in newborn infants and children is presented. Measurements of hip rotation and abduction are performed in a supine position. During examination, the foot is held in passive correction or induced weight-bearing position to compare the tibia, forefoot, and hindfoot alignment. Thirty-six newborn infants were examined randomly to establish standard measurements for newborns. The only statistically significant difference between male and female infants occurred in hip abduction and internal and external rotation of the hips during extension. This method of examination of the lower extremity in newborn infants is useful for routine evaluation in all children.

Characterization of activity and expression of isocitrate lyase in Mycobacterium avium and Mycobacterium tuberculosis.

Analysis by two-dimensional gel electrophoresis revealed that Mycobacterium avium expresses several proteins unique to an intracellular infection. One abundant protein with an apparent molecular mass of 50 kDa was isolated, and the N-terminal sequence was determined. It matches a sequence in the M. tuberculosis database (Sanger) with similarity to the enzyme isocitrate lyase of both Corynebacterium glutamicum and Rhodococcus fascians. Only marginal similarity was observed between this open reading frame (ORF) (termed icl) and a second distinct ORF (named aceA) which exhibits a low similarity to other isocitrate lyases. Both ORFs can be found as distinct genes in the various mycobacterial databases recently published. Isocitrate lyase is a key enzyme in the glyoxylate cycle and is essential as an anapleurotic enzyme for growth on acetate and certain fatty acids as carbon source. In this study we express and purify Icl, as well as AceA proteins, and show that both exhibit isocitrate lyase activity. Various known inhibitors for isocitrate lyase were effective. Furthermore, we present evidence that in both M. avium and M. tuberculosis the production and activity of the isocitrate lyase is enhanced under minimal growth conditions when supplemented with acetate or palmitate.

Abnormal folate metabolism and mutation in the methylenetetrahydrofolate reductase gene may be maternal risk factors for Down syndrome.

BACKGROUND: Down syndrome, or trisomy 21, is a complex genetic disease resulting from the presence of 3 copies of chromosome 21. The origin of the extra chromosome is maternal in 95% of cases and is due to the failure of normal chromosomal segregation during meiosis. Although advanced maternal age is a major risk factor for trisomy 21, most children with Down syndrome are born to mothers <30 y of age. OBJECTIVE: On the basis of evidence that abnormal folate and methyl metabolism can lead to DNA hypomethylation and abnormal chromosomal segregation, we hypothesized that the C-to-T substitution at nucleotide 677 (677C-->T) mutation of the methylenetetrahydrofolate reductase (MTHFR) gene may be a risk factor for maternal meiotic nondisjunction and Down syndrome in young mothers. DESIGN: The frequency of the MTHFR 677C-->T mutation was evaluated in 57 mothers of children with Down syndrome and in 50 age-matched control mothers. Ratios of plasma homocysteine to methionine and lymphocyte methotrexate cytotoxicity were measured as indicators of functional folate status. RESULTS: A significant increase in plasma homocysteine concentrations and lymphocyte methotrexate cytotoxicity was observed in the mothers of children with Down syndrome, consistent with abnormal folate and methyl metabolism. Mothers with the 677C-->T mutation had a 2.6-fold higher risk of having a child with Down syndrome than did mothers without the T substitution (odds ratio: 2.6; 95% CI: 1.2, 5.8; P < 0.03). CONCLUSION: The results of this initial study indicate that folate metabolism is abnormal in mothers of children with Down syndrome and that this may be explained, in part, by a mutation in the MTHFR gene.

Protection from 1-cyano-3,4-epithiobutane nephrotoxicity by aminooxyacetic acid and effect on xenobiotic-metabolizing enzymes in male Fischer 344 rats.

1-Cyano-3,4-epithiobutane (CEB), a naturally occurring nitrile derived from cruciferous plants, causes nephrotoxicity and increased renal glutathione (GSH) concentration in male F-344 rats. This CEB-induced nephrotoxicity is dependent on GSH conjugation and bioactivation. The objectives of the present study were to investigate the effect of CEB on several xenobiotic-metabolizing enzymes and to evaluate the effect of modulators of GSH transport and metabolism on CEB-induced nephrotoxicity and GSH concentration. Animals received 125 mg kg-1 CEB alone or following pretreatment with one of three selective inhibitors of GSH metabolism: acivicin, probenecid or aminooxyacetic acid. There were no significant alterations in epoxide hydrolase (EH), P-450, ethoxyresorufin O-deethylase (EROD) or pentoxyresorufin O-depentylase (PROD) enzyme activity, but renal glutamyl cysteine synthetase (GCS) activity was decreased at 12 and 24 h, as was renal glutathione S-transferase 4 h after CEB administration. Renal ECOD activity was also diminished at 24 h and at 12 and 24 h in liver. Aminooxyacetic acid (AOAA) abrogated the nephrotoxicity, the renal GSH-enhancing effect, and decreased GCS of CEB alone. These findings provide further evidence for the importance of GSH conjugation as a significant pathway in CEB metabolism and the role of a reactive thiol in nephrotoxicity and altered renal GSH.

Effects of oral administration of orotic acid on hepatic morphologic characteristics and serum biochemical variables in cats.

OBJECTIVE: To evaluate orotic acid (OA) as a possible etiologic factor in cats with idiopathic hepatic lipidosis (HL). ANIMALS: 20 clinically normal adult female cats. PROCEDURE: Cats were fed a control diet or a diet containing less protein. On day 1 of the control period, blood, urine, and liver biopsy specimens were obtained. Each cat was given an oral dose of water daily. On days 8, 15, and 22, blood and urine specimens were collected as on day 1. On day 29, liver, blood, and urine samples were obtained as on day 1. After a resting period of 30 to 60 days, cats were treated with orotic acid. Serum biochemical analyses, urinary OA-to-creatinine ratios, and liver biopsy specimens were evaluated. Cats were given OA orally (suspension or capsules) for 29 days. Sample collection and data obtained were identical to those described for the control period. RESULTS: Urinary OA-to-creatinine ratios were significantly higher in all treated cats, but ratios were significantly higher in those receiving OA in capsules than in those receiving OA in suspension. Diet or treatment did not alter hepatic biochemical or histologic variables significantly. However, 7 cats given the highest dose of OA in capsules developed azotemia, urolithiasis, and renal changes. CONCLUSIONS: Most concentrations of OA used in this study did not induce HL in cats during a 29-day period, but the highest dosage used did result in renal disease. CLINICAL RELEVANCE: Orotic acid does not appear to be involved in the genesis of HL in cats.

Urinary orotic acid-to-creatinine ratios in cats with hepatic lipidosis.

OBJECTIVE: To determine urinary orotic acid (OA) concentration and evaluate the urinary OA-to-creatinine ratio (OACR) in cats with hepatic lipidosis (HL). ANIMALS: 20 cats with HL and 20 clinically normal cats. PROCEDURE: Hepatic lipidosis was diagnosed on the basis of clinical signs, results of serum biochemical analyses, exclusion of other concurrent illness, and cytologic or histologic evaluation of liver biopsy specimens. Urine samples were collected from each cat and frozen at -20 C until assayed. Urine creatinine concentrations were determined, using an alkaline picrate method followed by spectrophotometric assay. Urine OA concentration was determined, using high-performance liquid chromatography. Minimum amount of detectable OA in feline urine was 1 microg/ml. Because of small interfering peaks near the base of the OA peak, the minimum quantifiable concentration of OA was determined to be 5 microg/ml. Urinary OACR was compared in both groups of cats. RESULTS: Differences in urinary OACR were not detected between clinically normal cats and cats with HL. Peaks were not detected for urinary OA in any of the 20 clinically normal cats. Of the 20 HL cats, 14 did not have detectable peaks for urinary OA. Of the 6 HL cats that had detectable urinary OA peaks, 3 had values of <5 microg/ml. CONCLUSIONS: Apparently, OACR does not increase significantly in cats with HL. CLINICAL RELEVANCE: Urinary OACR is not a useful diagnostic test for HL in cats.

Frostbite: an orthopedic perspective.

Frostbite injury to the extremities has the potential for disastrous effects. This review provides information valuable to the orthopedic surgeon to aid in the evaluation and treatment of frostbite. The pathophysiology and predisposing factors that provide a basic understanding of the nature of frostbite are discussed. Accepted and experimental imaging studies and treatment options are also reviewed. An effort is made to give the orthopedic perspective on each issue, providing a valuable resource for all orthopedic surgeons involved in the care of the patient with frostbite.

Hip osteotomy arthroplasty: ten-year follow-up.

We previously reported the initial success of combined osteotomy and arthroplasty of the hip for arthritis with femoral deformity. This technique has gained acceptance. We now report, for the first time, the ten year clinical and radiographic results with histology of 2 specimen. The osteotomies healed and the proximal femoral segment remained viable. One of three patients is symptom free without subsequent operative treatment. One of three patients had revision for acetabular loosening at eight years and biopsy of the proximal femur showed the proximal femoral segment to be viable. One of three patients had loosening of a macrofit bipolar prosthesis which required revision to total hip replacement at five years. Histology revealed viability of the proximal femur. All three patients are doing well at ten year follow-up. Based on the results of this study and current knowledge, the technique of osteotomy and arthroplasty for hip arthritis associated with femoral deformity is effective when combined with current techniques of ingrowth femoral component of total hip arthroplasty.

Analysis of DNA adduct, S-[2-(N7-guanyl)ethyl]glutathione, by liquid chromatography/mass spectrometry and liquid chromatography/tandem mass spectrometry.

Sensitive and specific isotope dilution liquid chromatography/mass spectrometry (LC/MS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS) methods were developed for the detection and quantitative analysis of S-[2-(N7-guanyl)ethyl]glutathione as a DNA adduct formed upon exposure of animals to carcinogenic 1,2-dihaloethanes. Separation and analysis were performed using microbore HPLC coupled in-line to an electrospray ionization triple quadrupole mass spectrometer. S-[2-(N7-guanyl)[2H4]-ethyl] glutathione was synthesized and used as internal standard. These methods provide structural confirmation of the adduct as well as quantitative analysis with the accuracy and precision necessary to measure biologically relevant levels in small tissue sample sizes (< 1 g). The sample detection limits in in vivo tissue extracts were 100 pg and 5 pg on-column for LC/MS and LC/MS/MS methods, respectively. Selected-ion monitoring mode was used to monitor the product ions of the doubly charged molecular ion. The application of these methods was demonstrated by measuring the DNA adduct levels in rat and fish samples after exposure to 1,2-dihaloethanes. The method has application in studies of DNA adduct formation as a biological marker of exposure to carcinogens and for environmental monitoring of 1,2-dihaloethanes.

Conversion of big endothelin-1 and characterization of its contractile effects on isolated human placental arteries.

OBJECTIVES: To study the conversion of human big endothelin-1 (bigET-1) to endothelin-1 (ET-1) and to characterize contractile ET-1 receptors in human placental arteries. METHODS: BigET-1 was incubated with artery membranes and the formation of ET-1 was investigated. ET-1 and bigET-1-induced contractile responses were studied in the absence or presence of the metalloprotease inhibitor phosphoramidon, the ET(A)-receptor antagonist BQ 123, or the ETB-receptor antagonists IRL 1038 and RES 701-1. RESULTS: The artery membranes hydrolysed bigET-1 to ET-1 through a partly phosphoramidon-sensitive pathway. The contractile responses to ET-1 and bigET-1 were similar, with pEC50% values of 8.1 +/- 0.2 and 7.8 +/- 0.1, respectively (NS; n = 17). Phosphoramidon decreased pEC50% for bigET-1-evoked contractions (p < 0.05; n = 8), without affecting the response to ET-1. A Schild plot of BQ 123 effects on ET-1 and bigET-1-induced contractions resulted in identical pA2 values and a slope of 0.56 +/- 0.2 and 0.47 +/- 0.01, respectively. IRL 1038 and RES 701-1 did not affect the contractile responses. CONCLUSION: BigET-1-evoked contractions in isolated human placental arteries depend on a rapid and metalloprotease-dependent hydrolytic conversion to ET-1, which in turn causes a, mainly ETA-receptor-mediated, contraction.