The effect of a non-denaturing detergent and a guanidinium-based inactivation agent on the viability of Ebola virus in mock clinical serum samples.

The 2014 Ebola outbreak in West Africa required the rapid testing of clinical material for the presence of potentially high titre Ebola virus (EBOV). Safe, fast and effective methods for the inactivation of such clinical samples are required so that rapid diagnostic tests including downstream analysis by RT-qPCR or nucleotide sequencing can be carried out. One of the most commonly used guanidinium - based denaturing agents, AVL (Qiagen) has been shown to fully inactivate EBOV once ethanol is added, however this is not compatible with the use of automated nucleic acid extraction systems. Additional inactivation agents need to be identified that can be used in automated systems. A candidate inactivation agent is Triton X-100, a non-denaturing detergent that is frequently used in clinical nucleic acid extraction procedures and has previously been used for inactivation of EBOV. In this study the effect of 0.1% and 1.0% Triton X-100 (final concentration 0.08% and 0.8% respectively) alone and in combination with AVL on the viability of EBOV (106 TCID50/ml) spiked into commercially available pooled negative human serum was tested. The presence of viable EBOV in the treated samples was assessed by carrying out three serial passages of the samples in Vero E6 cells (37°C, 5% CO2, 1 week for each passage). At the end of each passage the cells were observed for evidence of cytopathic effect and samples were taken for rRT-PCR analysis for the presence of EBOV RNA. Before cell culture cytotoxic components of AVL and Triton X-100 were removed from the samples using size exclusion spin column technology or a hydrophobic adsorbent resin. The results of this study showed that EBOV spiked into human serum was not fully inactivated when treated with either 0.1% (v/v) Triton X-100 for 10 mins or 1.0% (v/v) Triton X-100 for 20 mins (final concentrations 0.08% and 0.8% Triton X-100 respectively). AVL alone also did not consistently provide complete inactivation. Samples treated with both AVL and 0.1% Triton X-100 for 10 or 20 mins were shown to be completely inactivated. This treatment is compatible with downstream analysis by RT-qPCR and next generation sequencing.

Differential identification of Bacillus anthracis from environmental Bacillus species using microarray analysis.

AIMS: To determine whether microarray analysis could be employed for the differential identification of a range of environmental Bacillus sp. from four strains of Bacillus anthracis. METHODS AND RESULTS: Oligonucleotide probes were designed that were specific to virulence factor genes of B. anthracis (pag, lef and cap), the variable number tandem repeat region of the B. anthracis vrrA gene and to the 16S-23S rRNA intergenic transcribed spacer region (ITS) and pleiotropic regulator (plcR) regions of the Bacillus cereus subgroup species. Generic probes were also designed to hybridize with conserved regions of the 16S rRNA genes of Bacillus (as a positive control), Neisseria sp., Pseudomonas sp., Streptococcus sp., Mycobacterium sp. and to all members of the Enterobacteriaceae to allow simultaneous detection of these bacteria. Identification of B. anthracis was found to rely entirely on hybridization of DNA specific to regions of the pag, lef and cap genes. Cross-reaction was observed between B. anthracis and other Bacillus species with all the other Bacillus probes tested. Results obtained using microarray hybridizations were confirmed using conventional microbiological techniques and found to have very high comparability. CONCLUSIONS: Microarray-based assays are an effective method for the identification of B. anthracis from mixed-culture environmental samples without problems of false-positivity that have been observed with conventional PCR assays. SIGNIFICANCE AND IMPACT OF THE STUDY: Identification of environmental Bacillus sp. by conventional PCR is prone to potential for reporting false-positives. This study provides a method for the exclusion of such isolates.

Relation of serum 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) level to hematological examination results in veterans of Operation Ranch Hand.

The authors studied indices of hematologic function and exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in Vietnam War veterans of Operation Ranch Hand--the Air Force unit responsible for the aerial spraying of Agent Orange and other herbicides in Vietnam. The herbicides were contaminated with TCDD. The authors measured TCDD serum levels in 1987 or later and extrapolated the result to the time of service in Vietnam. The authors studied serum TCDD level in relation to red blood cell count, hemoglobin, hematocrit, mean corpuscular volume, white blood cell count, platelet count, and erythrocyte sedimentation rate at each of 4 physical examinations. Compared with veterans not involved in Operation Ranch Hand, those with the highest TCDD levels in Operation Ranch Hand had mean corpuscular volumes that were about 1% higher and platelet counts that were about 4% higher. These small increases were unlikely to be of clinical significance and may not have been caused by TCDD.

Serum dioxin and cancer in veterans of Operation Ranch Hand.

We studied cancer prevalence and exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin) in veterans of Operation Ranch Hand, the Air Force unit responsible for the aerial spraying of herbicides in Vietnam from 1962 to 1971. A comparison group of Air Force veterans who served in Southeast Asia during the same period and who were not involved with spraying herbicides was included. Comparison veterans were matched to Ranch Hand veterans on age, race, and military occupation. We measured dioxin in 1987 or 1992, extrapolated the result to the time of service in Southeast Asia, and assigned each Ranch Hand veteran to Background, Low, or High exposure categories. This study had low power to detect an effect for specific or rare cancers. The risk of cancer at sites other than the skin within 20 years of service was increased in the Low (odds ratio (OR) = 3.4, 95% confidence interval (CI) 1.5-8.0) and High (OR = 2.7, 95% CI 0.9-8.0) categories, but the pattern was inconsistent with another study, suggesting that the excess risk may not have been caused by dioxin exposure. Overall, we found no consistent evidence of a dose-response gradient and no significant increase in cancer risk in the High dioxin exposure category, the subgroup of greatest a priori interest.

Serum dioxin, chloracne, and acne in veterans of Operation Ranch Hand.

The possible relationship between exposure to Agent Orange and its contaminant, 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin), during the Vietnam War and chloracne was investigated. The index subjects were veterans of Operation Ranch Hand, the unit responsible for aerial herbicide spraying in Vietnam from 1962 to 1971. Other Air Force veterans who served in Southeast Asia during the same period, but who were not involved with spraying herbicides, served as comparisons. None of the Ranch Hand veterans were diagnosed with chloracne; therefore, we restricted our analyses to acne. We found no meaningful or consistent association between dioxin exposure and prevalence of acne without or with regard to anatomical location. These results suggested that exposure of Ranch Hand veterans to dioxin was insufficient for the production of chloracne or perhaps the exposure may have caused chloracne that resolved and was currently undetectable.

Leading the research initiative: the role of the nurse manager.

Outcomes research, an integral part of the nurse manager role, is important because it examines the end result of care and stimulates changes in nursing practice. Without outcomes research, the nurse manager lacks knowledge of what interventions have been tried or their resulting effectiveness. The move from episodic to a continuum of care emphasizes the need for continued research across all organizational boundaries. This paradigm shift questions traditions and requires finding new ways to plan and implement care. Nurse managers assure that they provide the leadership to develop and implement outcomes data to strengthen nursing's role on the health care team and to achieve positive patient outcomes.

The impact of medical libraries and literature on patient care in New Zealand.

This paper reports the results of a survey of 372 physicians working in regional hospitals in New Zealand. The survey was undertaken to determine if these physicians used their local medical library and what impact the literature obtained had on patient care. The 295 respondents (80.6% of the sample) expressed genuine interest in their library, with most visiting the library once a week or more. Patient care was ranked as the main reason for their visit; the second most important reason was keeping up-to-date on the latest research. Physicians indicated that information obtained at the library was used to confirm or change diagnoses, medications, or diagnostic tests, or to change advice given to patients. Information also was used to make decisions concerning early discharge, avoidance of hospital admission, or transfer of patients to another hospital. The survey also addressed the use of librarian services, sources of information used, and physicians' purchase of textbooks and journals. All comments by respondents were supportive of the library. The significance of libraries to clinical decision making, especially in small hospitals, is discussed.

Occupational therapy in long term psychiatry: what's new?

This paper summarizes the results of a survey of the role of occupational therapy in provincial psychiatric hospitals in Ontario. Occupational therapy was found to provide major programming in three areas: 1) functional skills assessment and training, 2) life skills assessment and training and, 3) self awareness and personal growth. Subjects used a variety of frames of reference, but models based upon Glasser's Reality Therapy predominated. A complementary relationship was found between occupational therapy and vocational and recreational services, with occupational therapy playing a clinical or therapeutic role, while vocational and recreational services provided normalizing experiences. A comparison of occupational therapy programming in the institutions surveyed, with programming reported in the recent occupational therapy literature indicated that techniques and media utilized paralleled those used in acute and community psychiatric programs.

Programming to meet the needs of the elderly in institutions: part II.

This paper explores the principles of humanistic psychology and their application in the care of elderly institutionalized people. Activities already used by occupational therapists which can enhance the self-actualization process are described and other activities which could be incorporated into activity programs are suggested. The influence of staff attitudes on residents' well-being is a vital factor in geriatric care and must be taken into account in any form of programming.

Programming to meet the needs of the elderly in institutions: part I.

A survey of 24 institutionalized elderly people was conducted to determine their present interests and activities in the institution and those prior to institutionalization. It was found that elderly women's former interests were maintained for the most part in the institutions except for homemaking, but that less opportunity was provided for elderly men to pursue their former interests. The types of activities and interests described by these subjects tend to correspond with the types of activities commonly included in programs designed by occupational therapists for elderly residents in institutions.

In Vivo Assay for the Synthesis of Hydroxyproline-rich Proteins.

Simple methods are described for following in vivo the rate of peptidylproline hydroxylation and for determining what proportion of the total proline incorporated into protein is hydroxylated.

Dependence of the activity of interpositus and red nucleus neurons on sensory input data generated by movement.

Cats performed flexion movements of the forearm, and the discharge of interpositus and red nucleus neurons was examined for relationships to the motion and to the EMG activity of agonist and antagonist muscles. It is shown that, over a wide range in the time course of the motion, the activity of the neurons is covariant with both the EMG and the movement parameters, in particular, the velocity. Also, the discharge of the interpositus neurons is modulated in phase with the velocity of passive movement. It is concluded that during fast, intentional movements, sensory input data generated by motion is a major determinant of the output of the interpositus nucleus. Furthermore, the results are consistent with the concept that this output provides a continuous modulation of spinal segmental mechanisms by way of the red nucleus and rubrospinal tract.

Relationships between sensory input, motor output and unit activity in interpositus and red nuclei during intentional movement.

The relationship between unit activity in interpositus (8 units) and red nuclei (11 units) and the EMG activity of the biceps during intentional elbow flexion movements was investigated by means of cross-correlation analysis. This analysis showed that there were long-lasting (200 msec) changes in the probability of EMG activity both before and after a single spike in neurons which covaried with the motor output. The dependence of the activity of these units on sensory inputs was investigated by (1) calculating the quantitative relationship between angular displacement and unit activity and (2) recording unit activity after the sensory input from peripheral afferents had been eliminated by dorsal rhizotomy.

Yes, you can teach clinical skills in the classroom!

This paper describes the use of videotaped interviews with volunteer patients as a method of teaching interviewing skills to first and second year occupational therapy students. It is hoped that this experience will enable students to conduct interviews in clinical or community settings with greater confidence and effectiveness.