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Therapy of advanced melanoma is changing dramatically. Following mutational and biological subclassification of this heterogeneous cancer, several targeted and immune therapies were approved and increased survival significantly. To facilitate further advancements through pre-clinical in vivo modeling, we have established 459 patient-derived xenografts (PDX) and live tissue samples from 384 patients representing the full spectrum of clinical, therapeutic, mutational, and biological heterogeneity of melanoma. PDX have been characterized using targeted sequencing and protein arrays and are clinically annotated. This exhaustive live tissue resource includes PDX from 57 samples resistant to targeted therapy, 61 samples from responders and non-responders to immune checkpoint blockade, and 31 samples from brain metastasis. Uveal, mucosal, and acral subtypes are represented as well. We show examples of pre-clinical trials that highlight how the PDX collection can be used to develop and optimize precision therapies, biomarkers of response, and the targeting of rare genetic subgroups., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)

OBJECTIVE Ruptured blister aneurysms remain challenging lesions for treatment due to their broad, shallow anatomy and thin, fragile wall. Historical challenges with both open microsurgical approaches and intrasaccular endovascular approaches have led to increased use of flow diversion for management of these aneurysms. However, the optimum paradigm, including timing of treatment, use of dual antiplatelet therapy, and number of flow-diverter devices to use remains unknown. The authors describe their experience with ruptured blister aneurysms treated with flow diversion at their institution, and discuss rates of rebleeding and number of devices used. METHODS All patients presenting with subarachnoid hemorrhage from a ruptured blister aneurysm and treated with Pipeline flow diversion were identified. Patient demographic data, clinical status and course, need for external ventricular drain (EVD), timing of treatment, and angiographic details and follow-up were recorded. RESULTS There were 13 patients identified (11 women and 2 men), and 4 had multiple aneurysms. Two aneurysms were treated on initial angiography, with average time to treatment of 3.1 days for the remainder, after discussion with the family and institution of dual antiplatelet therapy. Device placement was technically successful in all patients, with 2 patients receiving 2 devices and the remainder receiving 1 device. There was 1 intraoperative complication, of a wire perforation causing intracerebral hemorrhage requiring decompressive craniectomy. Three patients had required EVD placement for management of hydrocephalus. There was no rebleeding from the target lesion; however, one patient had worsening intraventricular hemorrhage and another had rupture of an unrecognized additional aneurysm, and both died. Of the other 11 patients, 10 made a good recovery, with 1 remaining in a vegetative state. Nine underwent follow-up angiography, with 5 achieving complete occlusion, 2 with reduced aneurysm size, and 2 requiring retreatment for aneurysm persistence or enlargement. There were no episodes of delayed rupture. CONCLUSIONS Pipeline flow diversion is a technically feasible and effective treatment for ruptured blister aneurysms, particularly in good-grade patients without hydrocephalus. Patients with a worse grade on presentation and requiring EVDs may have higher risk for bleeding complications and poor outcome. There was no rebleeding from the target lesion with use of a single device in this series.

Nursing leadership opportunities to improve quality and align resources in health care exist. An estimated 18% of United States gross domestic product is spent on health care delivery systems that produce poor outcomes. The purpose of this article was to describe how quality improvement and implementation science initiatives enhance outcomes using nursing leadership strategies that play an integral role in aligning key colleagues to drive the collaborative process. A critical appraisal of the literature was conducted, which supports the importance of evidenced-based practice improvement, collaborative change process, and professional role of nursing leadership. Limited evidence exists related to practice strategies for nursing leaders to implement sustainable change at the unit level for successful alignment of resources. Strategies based on Rogers' Diffusion of Innovation Theory are recommended to address the gap in the literature. The strategies aim to increase meaningful knowledge or the "why," create a tipping point, and implement sustainable change starting with the end in mind. Nurse leaders are a central component for driving alignment and implementing change at the unit level. Uses of the described evidenced-based strategies have implications for nursing practice, education, and scholarship.

Cerebral aneurysms pose a threat to patients because of their risk of rupture causing subarachnoid hemorrhage, and the goal of treatment is the exclusion of the aneurysm from the circulation to prevent bleeding (in the case of unruptured aneurysms) or rebleeding. This article analyzes the general technical factors associated with the endovascular treatment of cerebral aneurysms. It discusses issues with transarterial access; imaging of aneurysm size, morphology, and regional anatomy to determine the endovascular plan; the techniques for the major endovascular aneurysm devices; and periprocedural management issues to reduce potential treatment-related complications., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Objective: This study evaluates the role of endovascular therapy for middle cerebral artery (MCA) aneurysms and its evolution over time. We are reporting a large case series of 184 MCA aneurysms and comparative analysis of the literature., Methods: This was a retrospective review of a prospectively maintained database including all patients with MCA bifurcation aneurysms embolized between 2000 and 2013. There were 184 aneurysms in 166 patients, with 71 ruptured and 113 nonruptured aneurysms., Results: Stent assistance was required in 70 cases (38·0%) and 3 cases (1·6%) required 'Y' stenting. The initial rate of total aneurysm occlusion was 59·8% and at delayed follow up was 90·1%. Seven embolizations resulted in thrombo-embolic complications (3·8%), with no cases of aneurysm re-bleeding, wire perforations, or other hemorrhages. Three patients with ruptured aneurysms died of causes unrelated to the embolization, and none with nonruptured aneurysms died in the follow-up period (total mortality 1·6% at 30 days post-procedure). A total of seven aneurysms (3·9%) required retreatment with an average follow up of 41 months. Prior to dedicated aneurysm stents, 68·8% of patients underwent embolization with the remainder surgically treated. Following the introduction of aneurysm stents in 2002, 92·0% of MCA aneurysms treated were embolized., Conclusions: During the past decade we have seen a treatment paradigm shift in MCA aneurysm treatment from surgical treatment to endovascular treatment. Developments in 3D angiography, more compliant balloons, dedicated aneurysm stents, complex coils, and antiplatelet therapy regimes have led to this transition for safe and effective management of these patients.

Background: We sought to review the current literature with regards to future risks of hemorrhage following cerebral revascularization in Moyamoya disease (MMD)., Methods: We performed a comprehensive literature review using PubMed to inspect the available data on the risk of hemorrhage after revascularization in MMD., Results: In this review, we identify the risk factors associated with hemorrhage in MMD both before and after cerebral revascularization. We included proposed pathophysiology of the hemorrhagic risk, role of the type of bypass performed, treatment options, and future needs for investigation., Conclusions: The published cases and series of MMD treatment do show a risk of hemorrhage after treatment with either direct or indirect bypass both in the immediate as well as long-term future. While there are no discernible patterns in the rate of these hemorrhages, there is Class III evidence for the predictive effect of multiple microbleeds on preoperative imaging. Also, whereas revascularization, both direct and indirect, has been shown to reduce ischemic complications from MMD, there is not an association with the risk of hemorrhage after the procedure. Further studies need to be performed to help evaluate what the risk factors are and how to counsel patients as to the long-term outlook of this disease process.

Gas chromatography (GC) with a preparative fraction collector (PFC) has been used to facilitate the identification of a number of volatile impurities at major and minor percentage levels in a pharmaceutical matrix by nuclear magnetic resonance spectroscopy (NMR) and mass spectrometry (MS). The trapping process was optimized using liquid sorbents, and the impurities were trapped directly into a deuterated solvent. Challenges related to the pharmaceutical matrix were overcome by derivatization with boron trifluoride in methanol and extraction with heptane, producing the methyl esters of the carboxylic acid impurities and main component. GC coupled to atmospheric pressure chemical ionization mass spectrometry (APCI-MS) with a time-of-flight (TOF) detector was used to acquire accurate mass and isotopic data for the impurities, leading to the determination of their molecular formulas (MF). One dimensional (1D) and two-dimensional (2D) NMR experiments were also acquired to unambiguously determine the impurities' structure. The acquisition time of the latter experiments was minimized by using a high-resolution instrument equipped with a small (1.7 mm) cryogenic probe. The quality of the data was such that the structure of the impurities could be determined semiautomatically by using a computer-assisted structure elucidation (CASE) approach, even though the total amount of one of the isolated impurities was less than 60 nmol.

Object: The CO(2) laser has an excellent profile for use in neurosurgery. Its high absorption in water results in low thermal spread, sparing adjacent tissue. Use of this laser has been limited to line-of-sight applications because no solid fiber optic cables could transmit its wavelength. Flexible photonic bandgap fiber technology enables delivery of CO(2) laser energy through a flexible fiber easily manipulated in a handheld device. The authors examined and compared the first use of this CO(2) laser fiber to conventional methods for incising neural tissue., Methods: Carbon dioxide laser energy was delivered in pulsed or continuous wave settings for different power settings, exposure times, and distances to cortical tissue of 6 anesthetized swine. Effects of CO(2) energy on the tissue were compared with bipolar cautery using a standard pial incision technique, and with scalpel incisions without cautery. Tissue was processed for histological analysis (using H & E, silver staining, and glial fibrillary acidic protein immunohistochemistry) and scanning electron microscopy, and lesion measurements were made., Results: Light microscopy and scanning electron microscopy revealed laser incisions of consistent shape, with central craters surrounded by limited zones of desiccated and edematous tissue. Increased laser power resulted in deeper but not significantly wider incisions. Bipolar cautery lesions showed desiccated and edematous zones but did not incise the pia, and width increased more than depth with higher power. Incisions made without using cautery produced hemorrhage but minimal adjacent tissue damage., Conclusions: The photonic bandgap fiber CO(2) laser produced reliable cortical incisions, adjustable over a range of settings, with minimal adjacent thermal tissue damage. Ease of application under the microscope suggests this laser system has reached true practicality for neurosurgery.

Introduction: Recent developments in optical science and image processing have miniaturized the components required for confocal microscopy. Clinical confocal imaging applications have emerged, including assessment of colonic mucosal dysplasia during colonoscopy. We present our initial experience with handheld, miniaturized confocal imaging in a murine brain tumor model., Methods: Twelve C57/BL6 mice were implanted intracranially with 10(5) GL261 glioblastoma cells. The brains of 6 anesthetized mice each at 14 and 21 days after implantation were exposed surgically, and the brain surface was imaged using a handheld confocal probe affixed to a stereotactic frame. The probe was moved systematically over regions of normal and tumor-containing tissue. Intravenous fluorescein and topical acriflavine contrast agents were used. Biopsies were obtained at each imaging site beneath the probe and assessed histologically. Mice were killed after imaging., Results: Handheld confocal imaging produced exquisite images, well-correlated with corresponding histologic sections, of cellular shape and tissue architecture in murine brain infiltrated by glial neoplasm. Reproducible patterns of cortical vasculature, as well as normal gray and white matter, were identified. Imaging effectively distinguished between tumor and nontumor tissue, including infiltrative tumor margins. Margins were easily identified by observers without prior neuropathology training after minimum experience with the technology., Conclusion: Miniaturized handheld confocal imaging may assist neurosurgeons in detecting infiltrative brain tumor margins during surgery. It may help to avoid sampling error during biopsy of heterogeneous glial neoplasms, with the potential to supplement conventional intraoperative frozen section pathology. Clinical trials are warranted on the basis of these promising initial results.

Background: Assessing the impact of glioma location on prognosis remains elusive. We approached the problem using multivoxel proton magnetic resonance spectroscopic imaging (1H-MRSI) to define a tumor "metabolic epicenter", and examined the relationship of metabolic epicenter location to survival and histopathological grade., Methods: We studied 54 consecutive patients with a supratentorial glioma (astrocytoma or oligodendroglioma, WHO grades II-IV). The metabolic epicenter in each tumor was defined as the 1H-MRSI voxel containing maximum intra-tumoral choline on preoperative imaging. Tumor location was considered the X-Y-Z coordinate position, in a standardized stereotactic space, of the metabolic epicenter. Correlation between epicenter location and survival or grade was assessed., Results: Metabolic epicenter location correlated significantly with patient survival for all tumors (r2 = 0.30, p = 0.0002) and astrocytomas alone (r2 = 0.32, p = 0.005). A predictive model based on both metabolic epicenter location and histopathological grade accounted for 70% of the variability in survival, substantially improving on histology alone to predict survival. Location also correlated significantly with grade (r2 = 0.25, p = 0.001): higher grade tumors had a metabolic epicenter closer to the midpoint of the brain., Conclusions: The concept of the metabolic epicenter eliminates several problems related to existing methods of classifying glioma location. The location of the metabolic epicenter is strongly correlated with overall survival and histopathological grade, suggesting that it reflects biological factors underlying glioma growth and malignant dedifferentiation. These findings may be clinically relevant to predicting patterns of local glioma recurrence, and in planning resective surgery or radiotherapy.

In this historical review the authors examine the important developments that have led to the availability of laser energy to neurosurgeons as a unique and sometimes invaluable tool. They review the physical science behind the function of lasers, as well as how and when various lasers based on different lasing mediums were discovered. They also follow the close association between advances in laser technology and their application in biomedicine, from early laboratory experiments to the first clinical experiences. Because opinions on the appropriate role of lasers in neurosurgery vary widely, the historical basis for some of these views is explored. Initial enthusiasm for a technology that appears to have innate advantages for safe resections has often given way to the strict limitations and demands of the neurosurgical operating theater. However, numerous creative solutions to improve laser delivery, power, safety, and ergonomics demonstrate the important role that technological advances in related scientific fields continue to offer neurosurgery. Benefiting from the most recent developments in materials science, current CO(2) laser delivery systems provide a useful addition to the neurosurgical armamentarium when applied in the correct circumstances and reflect the important historical advances that come about from the interplay between neurosurgery and technology.

A simple, reliable and fast procedure for the simultaneous determination of residues of some common alkylating agents (AAs), such as mesylates, besylates, tosylates and sulfates, employed in drug synthesis, has been developed by in situ derivatization-headspace-gas chromatography-mass spectrometry. Pentafluorothiophenol is used as a derivatizing agent in different water/dimethyl sulfoxide ratios. Compared to former analytical procedures, this approach returns improvements in analysis time, selectivity, analyte stability and method sensitivity (LOD=0.11 microgg(-1) for methyl tosylate). The method exhibits low matrix dependence, excellent accuracy, precision (R.S.D.=2.8-10% range at 1 microgg(-1)) and robustness through the use of deuterated internal standards. Knowledge of the synthetic route allows a targeted approach to the determination of specific AAs since the procedure does not distinguish between acid species. The procedure was successfully applied to different pharmaceutical matrixes, and is particularly suitable for routine analysis with high sample throughput.

The ability of dopamine to induce apoptosis in a variety of cell types, including PC12 cells and neurons, has been well documented. Under non-reducing conditions, dopamine can be oxidized to semi-quinone and quinone species, which have the ability to arylate proteins and lead to the formation of covalent adducts. Potentially, it is the arylation of substrates critical to cell survival and/or the formation of toxic adducts which leads to the death observed after dopamine treatment. We have previously described the ability of a substituted monohydroxy-tetra-hydronaphthalene (DATN) to bind proteins that are susceptible to arylation by dopamine and related catecholamines. As DATN can prevent the covalent incorporation of dopamine into substrate molecules, we hypothesized that this compound could have a protective effect on cells that undergo apoptotic death in response to dopamine exposure. We report here that DATN prevents the dopamine-induced apoptotic death of PC12 cells in a dose-dependent manner. DATN did not prevent the oxidative stress associated with dopamine treatment, as lipid peroxide production was not influenced by DATN treatment. The ability of DATN to prevent dopamine-induced cell death was selective for this insult, as this compound did not influence the death of PC12 cells induced by hydrogen peroxide (H(2)O(2)). Consistent with this finding, DATN did not alter lipid peroxidation, nor oxidation of intracellular dichlorodihydrofluorescein subsequent to H(2)O(2) treatment. Consistent with a reduction in apoptotic death, the increase in caspase-3 activity associated with dopamine treatment was also prevented by DATN. These observations suggest that DATN may act to prevent one of the pathways linking dopamine and oxidative stress to caspase-3 activation. We propose that the inhibition of substrate arylation by the products of dopamine oxidation may provide a useful strategy for the prevention of dopamine-induced cell death.

Genetic, neurochemical, and environmental factors have been implicated in neurodegenerative disease, and a combination of these factors is likely responsible for disease onset and progression. Environmental toxicants implicated in Parkinson's disease include organic compounds, reactive oxygen species, metal ions and others. Exposure to a combination of environmental toxicants may produce a synergistic insult leading to neuronal death, even though levels of individual toxicants may be below detection by conventional methods. Rodent models of toxicant-induced neurodegeneration are hampered by the high resistance of these animals to many environmental toxicants. Extensive literature on aquatic toxicology and the high homology between many human and fish neurotrophic factors makefish a useful model for investigating environmental toxicants and neurodegeneration. Skin color in salmonids is under catecholaminergic control; pigment-containing melanophores aggregate when stimulated, resulting in paling. We demonstrate that lesions to nerves innervating melanophores prevent aggregation and produce dark skin color. The time course for return of skin color corresponds to neuronal regeneration, a neurotrophin-dependent event. Observations from this model system may be useful for predicting risks associated with environmental toxicants and nervous system integrity, and may have important implications for the identification of risk factors.

During a down-sizing of residency programs at a State University Medical School, hospital based residents' positions were eliminated. It was determined to find out the characteristics of the residents who graduated from the Laboratory Medicine Program, to compare women graduates with men graduates, and to compare IMGs with United States Graduates. An assessment of a 25 year program in laboratory medicine which had graduated 100 residents showed that there was no statistically significant difference by chi 2 analysis in positions (laboratory directors or staff), in certification (American Board of Pathology [and subspecialties], American Board of Medical Microbiology, American Board of Clinical Chemistry) nor in academic appointments (assistant professor to full professor) when the male graduates were compared with the female graduates or when graduates of American medical schools were compared with graduates of foreign medical schools. There were statistically significant associations by chi 2 analysis between directorship positions and board certification and between academic appointments and board certification. Of 100 graduates, there were 57 directors, 52 certified, and 41 with academic appointments. Twenty-two graduates (11 women and 11 men) attained all three.

Using the results of Epstein-Barr virus-specific immunofluorescence serology as the "gold standard," we found that the sensitivities of the five rapid test kits varied from 78 to 84% and specificities varied from 89 to 100%. Enzyme-linked immunosorbent assay-determined specific Epstein-Barr virus antibody profiles had a sensitivity and specificity of 98.6 and 95.5%, respectively.

The results of diagnostic testing for the detection of Chlamydial infections in Iceland during the years 1982 to 1994 were reviewed. During those 13 years 123,461 laboratory tests were performed in 101,574 examinations. These examinations were positive in 14,462 instances. The first diagnostic test to be introduced was cell culture in 1982. From then on the number of examinations and the number of positive examinations increased steadily until 1988, when positive examinations reached a peak at approximately 570 cases per 100,000 inhabitants. In 1990 a sharp decline in both the total number of examinations and positive results was observed. The percentage of positive examinations declined during the study period. In 1991 and 1992 the number of examinations, the number of positive examinations and the percentage of positive examinations increased but the number of positive tests declined again in 1993. In 1994 the polymerase chain reaction assay (PCR) replaced the much less sensitive Chlamydiazyme(R) assay and the number of positive examinations rose again although the number of tests declined. The dramatic reduction in prevalence experienced in Sweden does not seem to have taken place in Iceland. In Sweden a substantial effort was made to screen asymptomatic populations. In Iceland the screening of asymptomatic patients increased from the beginning of the study period until 1988 but declined thereafter. Screening of asymptomatic populations as well as contact tracing may be important for bringing about a significant reduction of the prevalence of sexually transmitted infections caused by Chlamydia trachomatis.

Infection with Borrelia burgdorferi, the etiologic agent of Lyme disease, is associated with an early and dominant humoral response to the spirochete's 23-kDa outer surface protein C (OspC). We have cloned and expressed OspC as a fusion protein in Escherichia coli and have shown that patient serum samples react with it in an enzyme-linked immunosorbent assay (ELISA) (S. J. Padula, A. Sampieri, F. Dias, A. Szczepanski, and R. W. Ryan, Infect. Immun. 61:5097-5105, 1993). Now we have compared the detection of B. burgdorferi-specific immunoglobulin M antibodies in 74 individuals with culture-positive erythema migrans by a whole-cell ELISA, immunoblot, and the recombinant OspC (rOspC) ELISA. Seventy-six negative controls were also studied. With all of the tests, there was a statistically significant association between the duration of disease and the frequency of a positive result. With the rOspC ELISA, the predictive value of a positive test was 100% and the predictive value of a negative test was 74%. Similar results were obtained with the whole-cell ELISA and with the immunoblot using as the source of test antigen a strain of B. burgdorferi which expresses abundant levels of OspC. We conclude that the use of rOspC in an ELISA is a convenient, readily automated, and easily standardized test for the serodiagnosis of early Lyme disease.

To determine the risk of acquiring Lyme disease or babesiosis from blood transfusion, serum was collected before and 6 weeks after patients received multiple transfusions during cardiothoracic surgery and antibodies to Borrelia burgdorferi and Babesia microti were measured. Of 155 subjects, 149 received 601 total units of packed red blood cells (PRBC) and 48 received 371 total units of platelets. No patient developed clinical or serologic evidence of Lyme disease; 1 (who received 5 units of PRBC) developed clinical and serologic evidence of babesiosis. The risk of acquiring Lyme disease from a transfused unit of PRBC was 0 (95% confidence interval [CI], 0-0.5%) and from a transfused unit of platelets was 0 (95% CI, 0-0.8%); the same risks for babesiosis were 0.17% (95% CI, 0.004%-0.9%) and 0 (95% CI, 0-0.8%), respectively. The risk of acquiring either Lyme disease or babesiosis from a blood transfusion in Connecticut is very low.

Background and Objectives: Single dose regimens have advantages in the treatment of STD. Azithromycin has unique pharmacokinetics that may make single dose regimens feasible. Treatment with a single 1 g dose of azithromycin was compared to 100 mg doxycycline twice daily for seven days., Study Design: This was a randomized third-party blinded study on 183 male patients, 176 of whom could be evaluated for efficacy., Results: Chlamydia trachomatis was cultured from 148 patients, 79 receiving azithromycin and 69 receiving doxycycline. Six patients receiving azithromycin had positive cultures on follow-up, four were known to have had sexual intercourse with infected partners. Fifty-one patients had gonorrhea; 28 were treated with azithromycin and 23 with doxycycline. Neisseria gonorrhoeae was eradicated from all patients except one receiving azithromycin. He denied sexual exposure during follow-up. Sixty patients were infected with Ureaplasma urealyticum, 35 were treated with azithromycin and 25 with doxycycline. Five patients in each group had positive cultures on follow up. Three patients receiving azithromycin and two receiving doxycycline were known to have had sexual exposure during follow-up., Conclusion: A single dose of azithromycin showed similar effectiveness as a 7-day regimen of doxycycline.

We have found that sera from patients with early stages of Lyme disease contain predominant immunoglobulin M reactivity to a major 23-kDa protein (p23) from Borrelia burgdorferi 2591 isolated in Connecticut. To characterize this immunodominant antigen, we cloned and sequenced p23 and found it to be 83% identical by nucleotide sequence and 75% identical by amino acid sequenced to pC (recently renamed OspC), an abundantly expressed protein on the outer surface of PKo, a European strain of B. burgdorferi (B. Wilske, V. Preac-Mursic, S. Jauris, A. Hofmann, I. Pradel, E. Soutschek, E.Schwab, G. Will, and G. Wanner, Infect. Immun. 61:2182-2191, 1993). In addition, immunoelectron microscopy localized p23 to the outer membrane, confirming that p23 is the strain 2591 homolog of OspC. The North American strain B31, commonly used in serologic assays for Lyme disease, does not express OspC. Northern (RNA) blot analysis detected low levels of ospC mRNA in B31, and DNA sequencing of the ospC gene from B31 revealed a 54-bp deletion in the upstream regulatory region, possibly accounting for the low transcriptional activity of ospC. The ospC coding region from B31 was cloned and antibody-reactive OspC was expressed in Escherichia coli. An immunoglobulin M enzyme-linked immunosorbent assay using recombinant OspC as the target antigen shows promise for the serodiagnosis of early stages of Lyme disease.

Polymerase chain reaction was used to detect herpes simplex virus (HSV) specific deoxyribonucleic acid (DNA) sequences in acute and convalescent cerebrospinal fluid (CSF) and brain tissue of a 78-year-old man and in CSF of a neonate who died of complications owing to herpes simplex virus encephalitis (HSVE). Polymerase chain reaction (PCR) was carried out for 35 cycles with a set of primers that bracketed a 92 base pair segment unique to the HSV DNA polymerase gene. Amplified DNA was electrophoresed on 3 percent agarose gel, blotted onto a nylon membrane, and probed with 32p-labeled oligonucleotide internal to the primers. The HSV specific DNA sequences were detected in the specimens from both patients. No HSV specific DNA was detected in CSFs from 20 patients with suspected Lyme disease or neurosyphilis. Polymerase chain reaction is a rapid and noninvasive technique for the diagnosis of HSVE.

To determine if antibodies to Borrelia burgdorferi persist after antibiotic treatment, we recalled 32 patients with Lyme disease from a primary care practice a mean of 16 months after treatment and analyzed initial and follow-up serum samples by ELISA and immunoblot assays. Of the eight patients whose initial serum specimens were positive for IgM antibody by ELISA, three had positive titers of IgM antibody at follow-up; of the 23 patients whose initial serum specimens were positive for IgG antibody by ELISA, 19 had positive titers of IgG at follow-up. Of the five patients whose initial serum specimens were positive for IgM antibody by immunoblot, two had positive titers of IgM antibody at follow-up; of the 30 patients whose initial serum specimens were positive for IgG antibody by immunoblot, 29 had positive titers of IgG antibody at follow-up. The bands on the IgG immunoblot remained remarkably constant during the period from analysis of the initial specimen to that of the follow-up specimen. Nine of the 32 patients had persistent or recurrent symptoms, and ELISA and immunoblot were not helpful for identifying these nine patients.

One hundred and eighty-two patients were enrolled in a randomized third-party blinded study to assess the efficacy and safety of azithromycin in the treatment of sexually transmitted diseases. Three regimens of azithromycin, including a single oral dose, were compared with a standard treatment with doxycycline. The patients were followed for four weeks. Efficacy was evaluated in 168 patients (113 azithromycin, 55 doxycycline). Fourteen patients had negative cultures or did not come for all follow-up visits. Of the 168, 138 were infected with Chlamydia trachomatis, 43 with Neisseria gonorrhoeae, and 45 with Ureaplasma urealyticum. Ninety-six per cent of patients with chlamydial infections and 92% of those with gonorrhoea were cured with azithromycin. Two patients infected with N. gonorrhoeae, four with C. trachomatis and six with U. urealyticum had positive cultures on follow-up visits after receiving azithromycin. Of these 11 patients with positive cultures on follow-up visits, seven (five with U. urealyticum and two with C. trachomatis) violated the protocol by having intercourse with infected individuals during the study. Azithromycin was very well tolerated; one patient complained of mild abdominal pain shortly after receiving the drug, seven patients complained of mild nausea and two patients had mild diarrhoea.

Currently, the method of choice for the laboratory diagnosis of Clostridium difficile disease is the detection of cytotoxin in stool filtrates by tissue culture. Since many hospital laboratories do not have tissue culture facilities, there is a need for a rapid test which is both sensitive and specific to diagnose C. difficile disease. A commercial latex agglutination was compared with the conventional cytotoxin tissue culture assay for the detection of C. difficile or its toxin(s) in fecal specimens. Of the 574 specimens evaluated, 111 were cytotoxin positive while 97 were positive by the latex agglutination test. There were 17 specimens positive by latex agglutination but negative by tissue culture assay. The overall sensitivity and specificity of the CDT latex test was 86.1 percent and 95.3 percent respectively. This rapid latex test can serve as an excellent screening procedure for the presence of C. difficile. Those specimens positive by the latex test should be further evaluated for the presence of cytotoxin by tissue culture.

Four hundred and seventy-three men and women at high risk for sexually transmitted disease were tested for the presence of Chlamydia trachomatis in the urethra or the endocervix. Four groups were involved in this multicenter study of two direct fluorescent-antibody microscopy tests, Kallestad Pathfinder and Syva Microtrak, compared with culture techniques. Results from the test sites indicated that there was no significant difference overall in the sensitivity and specificity of the two test kits. However, there was some interlaboratory variation seen in the sensitivity of the microscopy, but little difference in the specificity. Either kit could be an effective screening method for C. trachomatis in high-risk populations.

Thirty-two human isolates of enterococci were tested for antibiotic synergy by using penicillin and one of six aminoglycosides. Three methods were used: synergy screen, microdilution checkerboard, and time-kill curves. The synergy screen accurately predicted synergy for gentamicin-penicillin combinations, and this synergy was later confirmed by time-kill curves. The microdilution checkerboard method suffered from inherent variation, and agreement with time-kill curves ranged from 92% (twofold reduction in minimum inhibitory concentration) to 4.2% (fourfold reduction in minimum inhibitory concentration). We suggest that enterococci be screened for synergy (i.e., presence or absence of high-level resistance) by using the criterion of growth or no growth in the presence of 2,000 microgram of an aminoglycoside per ml. The microdilution checkerboard test for synergy is not recommended.

Fifty fecal specimens were tested by three methods, bacterial isolation, counterimmunoelectrophoresis, and tissue culture, for Clostridium difficile and its toxin. Ten specimens (20%) were positive by all three methods. An additional eight specimens were toxin positive only by counterimmunoelectrophoresis. Although counterimmunoelectrophoresis and tissue culture are of equivalent sensitivity, the additional dilution necessary for tissue culture assay may be critical when only small concentrations of toxin are present.

Among 270 CT scans of the thorax obtained over a 7-month period, four patients (1.5%) with calcified herniated thoracic disks were identified. Each of these patients presented with abnormal chest radiographs and had a CT examination for evaluation of suspected malignancy. None showed any signs or symptoms of thoracic spinal cord compression. The clinical significance of incidental thoracic disk protrusions is unknown. It may be that these patients are at risk for the later development of symptomatic disk disease.

Whole cells of Pseudomonas aeruginosa possess rhodanese activity. The enzyme can be released by rapidly resuspending the cells in cold Tris--HCl buffer. Approximately 95% of the rhodanese activity is released by cold shock. Release of the enzyme can be inhibited either by preincubating the cells with Mg2+ or by incorporating Mg2+ into the shocking buffer. The effect of Mg2+ can be reversed by washing the cells twice with buffer prior to cold shock. While rhodanese can be released from P. aeruginosa by cold shock, lactic dehydrogenase, a cytoplasmic enzyme, remains within the cell. Diazo-7-amino-1,3-napthalenedisulfonic acid, a compound which does not penetrate the cytoplasmic membrane, completely inactivated rhodanese and alkaline phosphatase, a periplasmic enzyme, whereas lactic dehydrogenase retained its full activity. These data suggest that rhodanese in P. aeruginosa, like alkaline phosphatase, is located distal to the cytoplasmic membrane in the periplasmic space. Electron micrographs also show that portions of the lipopolysaccharide outer membrane are shed from the cell during cold shock, while cells preincubated with Mg2+ did not release segments of their outer membrane.

Members of the genus Pseudomonas are widely implicated in human disease. Although most isolates are P. aeruginosa, there have been serious outbreaks of infection with other members of the genus. The susceptibility patterns of 5) Pseudomonas isolates are reported. It was notable that of the four antibiotics tested, uniform susceptibility was observed with minocycline.

The laboratory diagnosis of Mycoplasma pneumoniae is often difficult because of lengthy and complicated cultural methods and serological tests that may be both insensitive and nonspecific. In this study, 82 patients suspected of Mycoplasma pneumonia were cultured for M. pneumoniae, and their respiratory secretions were tested by a DNA probe for M. pneumoniae. The probe test was 100% sensitive and 98% specific compared to culture. This DNA probe, then, is an effective alternative method for the detection of M. pneumoniae in respiratory specimens.

Three commercial products and counterimmunoelectrophoresis were evaluated for their ability to detect microbial antigens of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in cerebrospinal fluid from 157 patients suspected of having meningitis. Thirty-four patients were diagnosed as having bacterial meningitis by culture, microscopy, or antigen detection. The overall results showed the following detection percentages; counterimmunoelectrophoresis, 76%; Phadebact CSF, 76%; Directigen, 82%, and Bactigen, 93%. The results with purified antigen revealed that latex agglutination was more sensitive than coagglutination, which in turn was more sensitive than counterimmunoelectrophoresis.

Clostridium difficile has been shown to be the major cause of antibiotic-associated gastroenteritis in both humans and experimental animals. During the past few years an increasing number of laboratories have attempted to detect, isolate, and identify this organism and its toxin from clinical samples. Direct visualization of C. difficile in patient specimens using immunofluorescent microscopy has been proposed. The major disadvantage of this method was its lack of specificity due to crossreaction with other clostridial species. Attempts to absorb the antisera with crossreacting strains also failed. Laboratory diagnosis of C. difficile in clinical specimens has relied on either culture using one or more selective media or on the detection of specific cytotoxin in stool filtrates. Until recently the cytotoxicity assay was the only procedure available for the routine detection of cytotoxin and, as a result, has limited this test to laboratories with access to tissue culture facilities. As a result, there has been much interest in the development of immunochemical methods for the detection of C. difficile toxins. We originally reported on the detection of C. difficile toxin in stool filtrates using counterimmunoelectrophoresis. We examined 140 fecal specimens submitted for C. difficile toxin assay by counterimmunoelectrophoresis, using both unabsorbed and absorbed antitoxin, tissue culture, and bacterial culture. Using tissue culture assay as the reference method, the sensitivity of counterimmunoelectrophoresis and counterimmunoelectrophoresis-absorbed was 100% and the specificity 63.0% and 77.5%, respectively. Enzyme immunosorbent assays for the detection of toxin A from C. difficile have also been reported, however, at the present time they do not appear to be as sensitive as the cytotoxicity assay for toxin B (cytotoxin).(ABSTRACT TRUNCATED AT 250 WORDS)

A micro broth-dilution method was developed for the antibiotic susceptibility testing of Bacteroides frgilis. Eighty strains of B. fragilis were tested against six antibiotics using the agar dilution test as the reference method. The microdilution test yielded results in 24 hours, and agreement with the reference test was satisfactory. Certain subspecies-specific patterns of antibiotic susceptibility were observed. However, larger numbers of subspecies should be evaluated before subspeciation of B. fragilis can be used as a reliable predictor of antibiotic susceptibility.

Chlamydia trachomatis has been shown to be a major cause of sexually transmitted diseases in the United States. An enzyme immunoassay (Abbot Laboratories) has been developed that detects chlamydial antigen directly in the urogenital specimens of patients. We have evaluated specimens from 1,074 patients belonging to one of three risk groups. Three swabs were collected from each patient--one each for Neisseria gonorrhoeae, chlamydia cell culture, and enzyme immunoassay. When compared with cell culture, the sensitivity and specificity of the enzyme immunoassay for symptomatic males and females attending a sexually transmitted disease clinic was 82% and 100%, and 91.3% and 95.0%, respectively. A moderate risk group, consisting of female patients seen at either urology or gynecology clinics for genitourinary symptoms was also evaluated. The sensitivity and specificity of the test on this group was 96% and 96.7%. A population of females at low risk were also screened for chlamydial infection. In this group, the sensitivity and specificity of the enzyme immunoassay was 89.3% and 93.2%, respectively. This rapid test is a highly specific and sensitive procedure for the detection of chlamydial antigen in genital specimens from high risk female patients as well as symptomatic males.

Eighty human serum specimens tested concomitantly by immunoblot and an enzyme-linked immunosorbent assay developed jointly at the University of Connecticut School of Medicine and the Connecticut Agricultural Experiment Station were used to evaluate three commercially available diagnostic products for Lyme borreliosis. The sources of the kits were Hillcrest Biologicals, Cypress, Calif.; Whittaker Bioproducts, Walkersville, Md.; and Cambridge Bioscience, Worcester, Mass. When compared with Western blot analysis, the sensitivities and specificities, respectively, for the diagnostic assays were as follows: Hillcrest Biologicals, 93 and 75%; Whittaker Bioproducts, 73 and 100%; Cambridge Bioscience, 89 and 100%; and University of Connecticut School of Medicine, 96 and 92%.

Multiple seawater sites in the northeastern United States, particularly Long Island Sound, and shellfish from Long Island Sound were sampled from April to November for 3 successive yr, 1983-1985. Hospitals in coastal and metropolitan areas of Connecticut were surveyed for the same 3-yr period, Vibrio vulnificus can be found in these waters during the summer months. The appearance of these virulent bacteria in both seawater and shellfish are a function of the water temperature; no V. vulnificus could be isolated until the temperature was approximately 17 degrees C. Although the risk of infection is small, as shown by isolation of this organism from patients, certain high-risk groups exist. Consumption of raw shell fish during the summer months should be discouraged in people with liver disease or patients on immunosuppressive therapy.

Forty-five isolates of gentamicin resistant Pseudomonas aeruginosa were evaluated for gentamicin-carbenicillin synergy. Only 9 percent (4/45) showed synergy. Of nine isolates with demonstrable zones around a gentamicin disc (8 to 12 mm), none showed a synergistic response. Effective treatment of gentamicin resistant Ps. aeruginosa with gentamicin and carbenicillin should not be assumed without additional test procedures.

Infectious mononucleosis (IM) is caused by the Epstein-Barr virus. Commonly used laboratory tests used for diagnosis of IM include a screening test based on the observation that horse erythrocytes are agglutinated by the Paul-Bunnell antibody found in the serum of patients with IM. This study evaluated two latex agglutination (LA) kits for IM, Monolatex (Wampole Laboratories) and Immunoscan-IM (American MicroScan) (formerly Monogen; Biokit, S.A.), and compared them with Monospot (Ortho Diagnostic Systems) results on 220 patient sera. Discrepancies in the three test results were resolved with complete Epstein-Barr virus antibody profiles. They indicated that any of the three kits tested can be successfully used as a screening test for IM. The advantage of the LA kits is that no differential absorption step is necessary. When discrepancies were resolved, sensitivity and specificity of both LA kits were greater than 93%.

A strain of Clostridium perfringens, type A, has been isolated from the intestine of a dog which died from parvovirus infection. This Clostridium strain produces a toxin which can be detected by counterimmunoelectrophoresis, using C. difficile antitoxin, and produces cytotoxicity in WI-38 cell culture. Cytopathology can be blocked by C. difficile antitoxin. Its role in canine parvovirus infection is unknown.

An automated agar plate streaker was evaluated. The Autostreaker mechanizes the agar plate streaking process by providing storage for plates, labeling and streaking one or more plates for either isolation or quantitation, and stacking in one of several racks for subsequent incubation. Results showed the Autostreaker to produce agar plates with well-separated colonies and accurate colony counts. A total of 1,930 clinical specimens were processed either in parallel with manual methods or solely by the Autostreaker. Technologist acceptance of machine-streaked plates was outstanding.