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.

An examination of calibration intervals required for accurately tracking blood pressure using pulse transit time algorithms.

Pulse transit time (PTT) is defined as the time it takes the blood pressure (BP) wave to propagate from the heart to a specified point on the body. After an initial BP measurement, PTT can track BP over short periods of time. This paper evaluates two PTT algorithms: Chen's and Poon's algorithm; two of the most cited works in the area. The criteria for evaluating them were: which was capable of best tracking changes in BP and which provided the longest time between subsequent BP measurements. These establish the suitability of the PTT method for practical applications, which has not been examined previously. Accuracy was evaluated using the Association of Advancement of Medical Instrumentation (AAMI) and the British Hypertension Society's (BHS) standards. Results show that Chen's algorithm is dependent on its lookup table at short intervals but remains accurate using a 6-min calibration interval, with r=0.96 and r(2)=0.98. Poon's algorithm fails when using a 2-min calibration interval, but is more capable of reflecting changes in BP. The short calibration interval and accuracy limit the usefulness of calculating BP using PTT. Therefore, neither of the algorithms can be recommended because of their shortcomings when estimating BP.

Dual polarisation interferometry characterisation of DNA immobilisation and hybridisation detection on a silanised support.

Dual polarisation interferometry is an analytical technique that allows the simultaneous determination of thickness, density and mass of a biological layer on a sensing waveguide surface in real time. We evaluated, for the first time, the ability of this technique to characterise the covalent immobilisation of single stranded probe DNA and the selective detection of target DNA hybridisation on a silanised support. Two immobilisation strategies have been evaluated: direct attachment of the probe molecule and a more complex chemistry employing a 1,2 homobifunctional crosslinker molecule. With this technique we demonstrate it was possible to determine probe orientation and measure probe coverage at different stages of the immobilisation process in real time and in a single experiment. In addition, by measuring simultaneously changes in thickness and density of the probe layer upon hybridisation of target DNA, it was possible to directly elucidate the impact that probe mobility had on hybridisation efficiency. Direct covalent attachment of an amine modified 19 mer resulted in a thickness change of 0.68 nm that was consistent with multipoint attachment of the probe molecule to the surface. Blocking with BSA formed a dense layer of protein molecules that absorbed between the probe molecules on the surface. The observed hybridisation efficiency to target DNA was approximately 35%. No further significant reorientation of the probe molecule occurred upon hybridisation. The initial thickness of the probe layer upon attachment to the crosslinker molecule was 0.5 nm. Significant reorientation of the probe molecule surface normal occurred upon hybridisation to target DNA. This indicated that the probe molecule had greater mobility to hybridise to target DNA. The observed hybridisation efficiency for target DNA was approximately 85%. The results show that a probe molecule attached to the surface via a crosslinker group is better able to hybridise to target DNA due to its greater mobility.

Design and fabrication of a silicon microreactor for DNA amplification.

A silicon microreactor consisting of an integrated heater, temperature sensor and thermal isolation chamber has been described. The thermal characteristics of the device have been studied by computer simulation and a rapid heating rate (20 degrees C--95 degrees C in less than 2 s) has been achieved. The fabrication process, consisting of microelectromechanical systems (MEMS) fabrication techniques has been established. The design features of this device, in particular the integrated heater and temperature sensor and the thermal isolation chamber allows fast heating/cooling rates and therefore enables efficient thermocycling suitable for DNA amplification.

The induction and detection in vitro of iNOS in the porcine basilar artery.

The expression of iNOS in vascular tissues has an adverse effect on vascular responses to vasoconstrictors and NO-mediated vasodilators. The development of a simple method for detecting the iNOS expression by functional means would be extremely useful. Here we describe a method for inducing iNOS in the porcine basilar artery followed by the detection of iNOS protein by immunocytochemical means and the characterisation of functional responses to U46619 and L-arginine. Porcine basilar arteries were treated with LPS (1, 10 and 100 microg/ml) for between 5 and 18 h at 37 degrees C. Inducible NOS protein was expressed in a concentration-dependent manner in the endothelial and smooth muscle cells after 5 h and persisted for 18 h. Vessels treated with LPS showed a time-dependent reduction in contractile function in response to U46619 (10 nM) reaching significance at the 18-h time point. Moreover, a similar time-dependent increase in the vasodilator response to exogenously applied L-arginine (30 microM) was observed at both 5- and 18-h time points. These effects of LPS at the 18-h time point were prevented by the incubation of vessels with dexamethasone (100 microM) in addition to LPS. The vasodilator response to L-arginine was prevented with the incubation with and in the presence of the inhibitor of inducible NOS, 1400W (10 microM) in addition to LPS. These results show that iNOS protein can be expressed in porcine cerebral arteries and that the iNOS is functional. The assessment of contractile function and responses to L-arginine using single concentrations is a rapid and effective method for establishing whether functional iNOS is present in porcine cerebral arteries.

Observations on the astrocyte response to a cerebral stab wound in adult rats.

The temporal and spatial distribution of immunocytochemically identified reactive astrocytes is described following a cerebral stab wound in adult rats. Different patterns of reactivity were observed in the cerebral cortex, the corpus callosum and the deep structures of the hemisphere. In the cerebral cortex, the zone of astrocytic reactivity was initially limited to the vicinity of the wound but spread with time to encompass the entire ipsilateral cortex, then regressed; in the deep structures only a spreading phase was observed and this was slower than in the cortex; reactivity in the corpus callosum was slight and always restricted to the immediate vicinity of the lesion. Reactive astrocytes were never observed in the hemisphere contralateral to the lesion. The reorganization of reactive astrocytes in the immediate vicinity of the lesion into a membrana gliae limitans accessoria was also observed. On the basis of these observations, the hypothesis is proposed that astrocytes respond primarily to the mechanical disruption consequent to injury and that the response promotes the restoration of the structural integrity of the lesioned tissue.

Deposition of scar tissue in the central nervous system.

Standard parasagittal lesions were placed stereotactically in the cerebral hemispheres of neonatal and adult rats in order to compare scarring in the immature and mature animal. Lesions were examined by light and electron-microscopy and immunofluorescence to study the astrocyte reaction, collagen deposition, and the formation of the basement membrane of the glia limitans. Normal mature scarring characterized by the deposition of collagen, astrocyte end-feet alignment over a glia limitans, and the permanent presence of mesodermal cells (fibroblasts and macrophages) in the core of the lesion, does not occur in wounds before 8-10 days post-partum (dpp). Instead there is no deposition of collagen, and only a transitory astrocyte response occurs with the formation of an interrupted glia limitans. These latter features disappear with time so that the wound is ultimately obliterated by the growth of axons and dendrites through the lesion. Mature scarring is attained over 8-12 dpp when increasing amounts of collagen are deposited and a continuous permanent glia limitans is formed. The acquisition of the mature response to injury from 8-12 dpp may be correlated with the presence of increasing titres of a fibroblast growth factor (FGF), derived from autolytic digestion of injured brain tissue. We have investigated FGF activity using a 3 T 3 fibroblast tissue culture assay to detect mitogenic activity in brain extracts from rats lesioned at different ages and from leukodystrophic mice which have no myelin. Our results show that high titres of FGF are present in the developing brain long before myelination commences, and that normal levels of FGF are found in the brains of leukodystrophic mice which have no myelin. Scarring in brain lesions in these mutants is quite normal.

Predicting risk of ischemic heart disease and cerebrovascular disease from systolic and diastolic blood pressures.

The relative importance of systolic versus diastolic blood pressure in predicting risk of ischemic heart disease or cerebrovascular disease is controversial. Since 1948 we have observed in the Manitoba Study 3983 men (most between 25 to 34 years old at entry), in whom risk of both diseases was determined using the multiple logistic model. Systolic and diastolic blood pressures after adjustment for age and body weight were compared at entry and at four other examinations during the follow-up period. When both blood pressures were considered together, a stronger association with cerebrovascular disease was found for systolic compared to diastolic blood pressure at entry and at most of the other examinations. For ischemic heart disease, diastolic pressure showed a stronger association at the earlier examinations, whereas systolic pressure was more important when the majority of the cohort was between 40 to 50 years of age. In middle-aged men the general concept that diastolic is more important than systolic is not justified for cerebrovascular disease or for ischemic heart disease.