Real-Time Duplex Applications of Loop-Mediated AMPlification (LAMP) by Assimilating Probes

Isothermal nucleic-acid amplification methods such as Loop-Mediated isothermal AMPlification (LAMP) are increasingly appealing alternatives to PCR for use in portable diagnostic system due to the low cost, weight, and power requirements of the instrumentation. As such, interest in developing new probes and other functionality based on the LAMP reaction has been intense. Here, we report on the development of duplexed LAMP assays for pathogen detection using spectrally unique Assimilating Probes. As proof of principle, we used a reaction for Salmonella enterica as a model coupled with a reaction for λ-phage DNA as an internal control, as well as a duplexed assay to sub-type specific quarantine strains of the bacterial wilt pathogen Ralstonia solanacearum. Detection limits for bacterial DNA analyzed in individual reactions was less than 100 genomic equivalents in all cases, and increased by one to two orders of magnitude when reactions were coupled in duplexed formats. Even so, due to the more robust activity of newly available strand-displacing polymerases, the duplexed assays reported here were more powerful than analogous individual reactions reported only a few years ago, and represent a significant advance for incorporation of internal controls to validate assay results in the field.     

Estimating signal strengths in the design of an indoor wireless network

One issue in the design and implementation of a wireless local area network is the selection of access point (AP) locations. Proper AP placement is necessary to provide adequate signal coverage and also to minimize cochannel coverage overlap. The impact of incorrect placement of APs is significant. Placing APs too far apart can lead to gaps in coverage. On the other hand, placing the units too close together leads to excessive cochannel coverage overlap, degrading system performance. Currently, AP placement involves a "trial and error" technique. When a technician tests the effect of moving an AP from one location to another, it is necessary to spend considerable time manually measuring signal strengths in order to determine how this move affects the AP's coverage area. In this letter, we describe a procedure for estimating the coverage areas of relocated APs. The procedure can be used as part of a manual design process or as part of an automated design tool.     

Alteration of Sertoli cell differentiation in the presence of carcinoma in situ in human testes

PURPOSE: We investigated Sertoli cells in testicular biopsies with carcinoma in situ (CIS) in respect to cytokeratin expression to elucidate the status of Sertoli cell differentiation adjacent to CIS in human testes. Cytokeratin 18 intermediate filaments indicate a state of undifferentiation usually observed in Sertoli cells of prepubertal testes. MATERIALS AND METHODS: 29 testicular biopsies presenting CIS were investigated by means of immunohistochemistry, using a polyclonal antibody against placental-alkaline phosphatase to detect CIS cells and a monoclonal antibody against human cytokeratin 18 to show expression of cytokeratin 18 intermediate filaments in Sertoli cells. RESULTS: All tubules bearing CIS showed positive cytokeratin expression of Sertoli cells if tubules were devoid of normal germ cells. However, a total of 13 specimen revealed CIS cells together with normal germ cells. In the presence of CIS cells together with round or elongated spermatids, adjacent Sertoli cells did not express cytokeratin immunoreactivity. In the case of combined presence of CIS and spermatogonia and primary spermatocytes, Sertoli cells could be found either immunopositive or immunonegative, and were positive in tubules with CIS and spermatogonia only. CONCLUSIONS: Sertoli cells associated with CIS cells undergo a process of dedifferentiation, seen by the re-expression of cytokeratin intermediate filaments. We suggest that this dedifferentiation results in a loss of Sertoli cell function and leads to a cessation of spermatogenic activity.     

The murine immune response to the male-specific antigen mouse testicular cytochrome c

Male and female A/J mice were examined for their ability to elicit T lymphocyte and antibody (Ab) responses to the male-specific Ag, mouse testicular cytochrome c (Mt cyt). T lymphocytes from both male and female mice primed in vivo responded to the Ag in in vitro proliferation assays, and the dose-response curves were statistically indistinguishable. In addition, similar levels of Ab to Mt cyt were observed in immunized male and female mice. The B cells producing the Ab had switched isotypes to IgG1 and IgG2a, indicating that the self-reactive T helper (Th) cells in male mice were functional. Thus, male mice do not appear to be immunologically tolerant to Mt cyt, at least at the Th and B lymphocyte levels. No evidence for disease was found in male mice primed with Mt cyt. Major histocompatibility complex (MHC) class II-positive antigen-presenting cells are present in the testes and these were shown in vitro to process and present Mt cyt to a T cell hybridoma specific for the synthetic peptide Mt cyt 93-104. However, the hybridoma was not activated in the absence of exogenous Mt cyt 93-104 or Mt cyt, indicating that endogenous Mt cyt is not normally processed in sufficient quantity to effectively load MHC class II molecules with this particular Mt cyt-derived peptide. Notwithstanding any immunologic privilege of the testes, the lack of tolerance to Mt cyt and its failure to elicit an autoimmune disease could extend from the low levels of processed Mt cyt Ag available for T cell recognition. The T cell response elicited by Mt cyt contrasts the lack of response to mouse somatic cytochrome c which differs from Mt cyt at 13 amino acid residues and is expressed in most tissues and at higher levels.     

An epidemic of roller-blade injuries in children

Roller blading is a new and increasingly popular leisure activity in many countries. We reviewed 110 consecutive patients with roller-blade injuries between 1 January and 30 June 1996. The patients ranged from 4 to 14 years in age (mean 6.5 years). Eighty-three (75.4%) sustained injuries to the upper limb and 27 (24.5%) injured the lower limb. Fifty-six patients, were girls and 54 were boys. Of the 110 patients, 79 (72.7%) sustained fractures, 28 (25.4%) soft tissue injuries and 3 (2.7%) dislocations. Eighty-three (75.4%) of the patients wore no protective equipment on the limbs. Four months following injury 103 (93.6%) patients were fully recovered. The mean duration of school absence was 3 days. Subsequently 101 children returned to using roller-blades following injury. Seventy-three (66.3%) of these now use protective equipment. We found that injuries were unrelated to age or duration of roller-blading experience or to the brand-name of roller blades used, and that most of our patients wore no protective equipment at the time of injury.     

Intralesional radiolabeled human monoclonal IgM in human tumor xenografts

Sensory-perceptual abnormalities in people with autism are discussed from two perspectives: published firsthand accounts and existing psychological research evidence. A range of abnormalities, including hyper- and hyposensitivity, sensory distortion and overload, and multichannel receptivity and processing difficulties, are described in firsthand accounts and frequently portrayed as central to the autistic experience. A number of dangers are inherent in uncritically accepting these accounts at face value and in any wider generalization to the autistic population as a whole. Evidence from clinical studies suggests that unusual sensory responses are present in a majority of autistic children, that they are manifested very early in development, and that they may be linked with other aspects of autistic behavior. In addition, experimental studies using a range of indices have found evidence of unusual responses to sensory stimuli in autistic subjects. However the clinical and experimental research to date suffers from serious methodological limitations and more systematic investigation is warranted. Key issues for future psychological research in the area are identified.     

Implicit theories and semantic similarities: Reply to Marsh and Groves (1987).

The role of implicit theories in behavioral judgments continues to be a source of debate. In a study exploring the factorial validity of student ratings of instruction, Cadwell and Jenkins (1985) suggested that students' implicit theories of instructor behavior may account for the robust factor structure underlying student ratings. They argued that students rely on the semantic similarity of the rating items, inflating the observed relations among the items. Marsh and Groves cite methodological and conceptual difficulties with the Cadwell and Jenkins study, but most of these difficulties may arise from Marsh and Groves's misconceptions about the role of implicit theories and semantic similarities and, more generally, the cognitive processes underlying judgment tasks. I review these issues and address various points of Marsh and Groves's critique. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Contamination of facial memory through exposure to misleading composite pictures.

In Exp I, 46 male and 51 female undergraduates witnessed a filmed incident and later described the target. Results show that Ss who observed a composite containing misleading information (either incorrect hair or an added moustache) were significantly more likely to misreport hairstyle and the presence of a moustache than those who did not. In Exp II, 210 17–65 yr old Ss observed a filmed incident and were tested for their recall and recognition of the target either immediately or after delays of 2 or 7 days. Significant bias, consistent with the content of the misleading composite, was present for both cued recall and choice of mug shot in a photo spread. The effect did not increase over delay but was greater when the composite was seen just prior to recall than immediately after the incident. Implications concerning interference with the memory of the witness who produced the composite and for other witnesses exposed to the misleading composite are discussed. (33 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The role of benchmarking for yardstick competition

With the increasing interest in yardstick regulation, there is a need to understand the most appropriate method for realigning tariffs at the outset. Benchmarking is the tool used for such realignment and is therefore a necessary first-step in the implementation of yardstick competition. A number of concerns have been raised about the application of benchmarking, making some practitioners reluctant to move towards yardstick based regimes. We assess five of the key concerns often discussed and find that, in general, these are not as great as perceived. The assessment is based on economic principles and experiences with applying benchmarking to regulated sectors, e.g. in the electricity and water industries in the UK, The Netherlands, Austria and Germany in recent years. The aim is to demonstrate that clarity on the role of benchmarking reduces the concern about its application in different regulatory regimes. We find that benchmarking can be used in regulatory settlements, although the range of possible benchmarking approaches that are appropriate will be small for any individual regulatory question. Benchmarking is feasible as total cost measures and environmental factors are better defined in practice than is commonly appreciated and collusion is unlikely to occur in environments with more than 2 or 3 firms (where shareholders have a role in monitoring and rewarding performance). Furthermore, any concern about companies under-recovering costs is a matter to be determined through the regulatory settlement and does not affect the case for using benchmarking as part of that settlement.