Non-quantitative Techniques Research Articles

Major histocompatibility complex class I in the horse (Equus caballus) placenta during pregnancy and parturition

BackgroundMajor histocompatibility protein class I (MHC-I) is believed to be expressed in the horse allantochorion only in limited areas at limited times. However, its expression has only been investigated in early pregnancy with non-quantitative techniques that cannot reliably detect small amounts of protein. ObjectiveTo quantify the relative expression of MHC-I in the allantochorion and endometrium during days 90–240 of pregnancy (PREG), parturition with physiological delivery of fetal membranes (PHYS), and parturition with retention of these membranes (FMR). Also, to visualize protein expression and determine whether classical or non-classical MHC-I mRNA is expressed. AnimalsHeavy draft horses. SettingPREG horses (n = 12) were sampled postmortem at a slaughterhouse. PHYS (n = 6) and FMR (n = 5) horses were sampled at farms in the vicinity of Olsztyn, Poland. MethodsFor relative quantification of MHC-I, western blotting with densitometry was used. To visualize MHC-I, immunohistochemistry was used. For mRNA identification, RT-PCR was performed. ResultsAlthough the quantity of MHC-I was lower during PREG than parturition, it was present in the allantochorion and endometrium during PREG. During parturition, MHC-I expression was upregulated in the allantochorion (PHYS vs. PREG: 2.7-times higher, 95% confidence interval, 1.3- to 5.7-times higher; FMR vs. PREG: 3.2-times higher, 95% confidence interval, 1.5- to 6.7-times higher). At parturition, staining for MHC-I was detected in the microcotyledons. Classical and non-classical MHC-I were expressed in both tissues during PREG, PHYS, and FMR. ConclusionMHC-I protein is present in the horse allantochorion and endometrium for at least the first two-thirds of pregnancy and at parturition.

Minimally Invasive Wearable Patch Device for Measurement of Interstitial Fluid Content

Introduction Edema, which is the medical term of swelling, is a condition in which excess interstitial fluid derived from a capillary lumen accumulates in a subcutaneous tissue. For example, edema can be caused daily by the effect of gravity mainly in lower limb of a healthy person during prolonged sitting on a chair. Such daily edema often accompany pain and languor, resulting in reducing quality of life. On the other hand, there is serious edema attributed from circulatory diseases such as cardiac arrest, myocardial infraction, expansion type cardiomyopathy, arteriosclerosis et al. The position where edema develops varies depending on the kind of primary disease. Conventionally, edema is diagnosed by non-quantitative technique such as direct observation of appearance or confirmation of an indent in the swollen skin after following finger pressure. Therefore, it is necessary to develop the devices which enable quantitative evaluation of the degree of edema in the local site of body not only for monitoring daily edema of healthy people, but also diagnosis of the patients with circulatory diseases and monitoring the effect of therapeutic treatment of the diseases. The electrical technique to apply an electric current to the living body through the skin is effective to quantify physiological information in a non- or low-invasive manner. In this research, we examined the relationship between interstitial water content and its electric resistance by applying the electric current to a self-made human body model. Experiments Surface structure of a human body is composed of epidermal layer which is outermost layer of the skin, dermic layer, and subcutaneous tissue. Edema is caused by accumulation of the excess interstitial fluid in dermic layer and subcutaneous tissue that are under the epidermis layer. In this research, we constructed a human body model by laminating a porcine-derived epidermal layer on a surface of a gelatin hydrogel which mimics a dermic layer. The gelatin hydrogel was prepared as follows. The gelatin powder was dissolved in a heated 1 wt% saline solution. The gelatin solution was poured into a mold and cooled to room temperature to facilitate gelation of the solution. Two kinds of gelatin hydrogels with different water content were prepared. The one included 70 wt% of water in the gel to simulate the dermic layer of healthy person, and the other was 80 wt% to simulate extreme swelling condition. Then, the electric current was applied to this model to examine the relationship between the water content of gelatin hydrogel and its electric resistance. Result and discussion Relationship between the water content of the gelatin hydrogel and its electric resistance is shown in Figure 1. Horizontal axis of Figure 1 represents the water content of gelatin hydrogel, and vertical axis the ratio of change in electric resistance in which 1 means the standardized electric resistance of the gel with 70 wt% of water content. From this result, we found that the electric resistance decreased with increase in water content of the gelatin hydrogel. From this, it was suggested that there is possibility to determine the degree of swelling in the body from the change of electric resistance by applying electric current to the body. Figure 1

Gene rearrangement study for minimal residual disease monitoring in children with acute lymphocytic leukemia

ObjectiveTo detect markers for minimal residual disease monitoring based on conventional polymerase chain reaction for immunoglobulin, T-cell receptor rearrangements and the Sil-Tal1 deletion in patients with acute lymphocytic leukemia. MethodsFifty-nine children with acute lymphocytic leukemia from three institutions in Minas Gerais, Brazil, were prospectively studied. Clonal rearrangements were detected by polymerase chain reaction followed by homo/heteroduplex clonality analysis in DNA samples from diagnostic bone marrow. Follow-up samples were collected on Days 14 and 28-35 of the induction phase. The Kaplan-Meier and multivariate Cox methods were used for survival analysis. ResultsImmunoglobulin/T-cell receptor rearrangements were not detected in 5/55 children screened (9.0%). For precursor-B acute lymphocytic leukemia, the most frequent rearrangement was IgH (72.7%), then TCRG (61.4%), and TCRD and IgK (47.7%); for T-acute lymphocytic leukemia, TCRG (80.0%), and TCRD and Sil-Tal deletion (20.0%) were the most common. Minimal residual disease was detected in 35% of the cases on Day 14 and in 22.5% on Day 28-35. Minimal residual disease on Day 28-35, T-acute lymphocytic leukemia, and leukocyte count above 50 x 109/L at diagnosis were bad prognostic factors for leukemia-free survival in univariate analysis. Relapse risk for minimal residual disease positive relative to minimal residual disease negative children was 8.5 times higher (95% confidence interval: 1.02-70.7).ConclusionImmunoglobulin/T-cell receptor rearrangement frequencies were similar to those reported before. Minimal residual disease is an independent prognostic factor for leukemia-free survival, even when based on a non-quantitative technique, but longer follow-ups are needed.

Quantitative sectioning and noise analysis for structured illumination microscopy

Structured illumination (SI) has long been regarded as a nonquantitative technique for obtaining sectioned microscopic images. Its lack of quantitative results has restricted the use of SI sectioning to qualitative imaging experiments, and has also limited researchers’ ability to compare SI against competing sectioning methods such as confocal microscopy. We show how to modify the standard SI sectioning algorithm to make the technique quantitative, and provide formulas for calculating the noise in the sectioned images. The results indicate that, for an illumination source providing the same spatially-integrated photon flux at the object plane, and for the same effective slice thicknesses, SI sectioning can provide higher SNR images than confocal microscopy for an equivalent setup when the modulation contrast exceeds about 0.09.

The Use Of Capital Budgeting Techniques In Businesses: A Perspective From The Western Cape

<p class="MsoNormal" style="text-align: justify; margin: 0in 0.5in 0pt; tab-stops: 1.0in;"><span style="font-family: Times New Roman;"><span style="font-size: 10pt; mso-fareast-font-family: Batang;" lang="EN-ZA">Capital budgeting is one of the most important areas of financial management. There are several techniques commonly used to evaluate capital budgeting projects</span><span style="font-size: 10pt;" lang="EN-ZA"> </span><span style="font-size: 10pt; mso-fareast-font-family: Batang;" lang="EN-ZA">namely the payback period, accounting rate of return, present value and internal rate of return and profitability index</span><span style="font-size: 10pt;" lang="EN-ZA">.</span><span style="font-size: 10pt; mso-fareast-font-family: Batang;" lang="EN-ZA"> Recent studies highlight that financial managers worldwide favor methods such as the internal rate of return (IRR) or non-discounted payback period (PP) models over the net present value (NPV), which is the model academics consider superior.</span><span style="font-size: 10pt;" lang="EN-ZA"> In particular this research focused on small, medium and large businesses and investigated a number of variables and associations relating to capital budgeting practices in businesses in the Western Cape province of South Africa. The results revealed that payback period, followed by net present value, appears to be the most used method across the different sizes and sectors of business. It was also found that 64% of businesses surveyed used only one technique, while 32% of the respondents used between two to three different types of techniques to evaluate capital budgeting decisions. The findings show that the more complicated methods such as IRR and NPV are most favored by the large businesses as compared to the small businesses. The majority of the respondents believed that project definition was the most important stage in the capital budgeting process. Implementation stage appeared to be the most difficult stage for the manufacturing sector whereas Project definition, Analysis and selection and Implementation were generally rated as being the difficult stages by the retail sector. Project definition and Analysis and selection </span><span style="font-size: 10pt; mso-ansi-language: EN-GB;" lang="EN-GB">were found to be the most difficult stages by the service sector.</span><span style="font-size: 10pt;" lang="EN-ZA"> Most businesses used the cost of bank loan as a basis in capital budgeting and</span><span style="font-size: 10pt; mso-ansi-language: EN-GB;" lang="EN-GB"> more than two thirds of respondents used non-quantitative techniques to consider risk when making a decision on investing in fixed assets. </span></span><span style="font-size: 10pt;" lang="EN-ZA"></span></p>

Comparison of different promoter methylation assays in breast cancer.

Background: Promoter hypermethylation has emerged as a promising cancer biomarker. Currently, a large variety of quantitative and non-quantitative techniques is used to measure methylation in clinical specimens. Here we directly compared three commonly used methylation assays and assessed the influence of tissue fixation, target sequence location and the amount of DNA on their performance. Methods: We used Methylation-Specific PCR (MSP), Quantitative Multiplex MSP (QM-MSP) and Methylation-Specific Multiplex Ligation-dependent Probe Amplification (MS-MLPA) to compare methylation of CCND2, SCGB3A1, RARB and RASSF1 on DNA from 40 breast carcinomas. Results: A comparison between MSP and QM-MSP on the same samples showed a high discrepancy: 20% of tumors that showed no methylation in MSP gave >10% methylation in QM-MSP. In contrast, QM-MSP correlated strongly with MS-MLPA when targeting the same sequence in DNA from paraffin embedded as well as fresh frozen tissue. This correlation declined when target sequences were non-overlapping. In titration experiments, MSP and MS-MLPA performed robust with 10 ng of DNA, while QM-MSP was at least ten-fold more sensitive. Conclusion: Despite the difference in molecular basis, QM-MSP and MS-MLPA showed moderate to strong correlations. In contrast, there was a poor concordance between either of these techniques and non-quantitative MSP. For biological samples with scarce DNA, QM-MSP is the method of choice.

Comparison of Different Promoter Methylation Assays in Breast Cancer

Background: Promoter hypermethylation has emerged as a promising cancer biomarker. Currently, a large variety of quantitative and non-quantitative techniques is used to measure methylation in clinical specimens. Here we directly compared three commonly used methylation assays and assessed the influence of tissue fixation, target sequence location and the amount of DNA on their performance.Methods: We used Methylation-Specific PCR (MSP), Quantitative Multiplex MSP (QM-MSP) and Methylation-Specific Multiplex Ligation-dependent Probe Amplification (MS-MLPA) to compare methylation of CCND2, SCGB3A1, RARB and RASSF1 on DNA from 40 breast carcinomas.Results: A comparison between MSP and QM-MSP on the same samples showed a high discrepancy: 20% of tumors that showed no methylation in MSP gave >10% methylation in QM-MSP. In contrast, QM-MSP correlated strongly with MS-MLPA when targeting the same sequence in DNA from paraffin embedded as well as fresh frozen tissue. This correlation declined when target sequences were non-overlapping. In titration experiments, MSP and MS-MLPA performed robust with 10 ng of DNA, while QM-MSP was at least ten-fold more sensitive.Conclusion: Despite the difference in molecular basis, QM-MSP and MS-MLPA showed moderate to strong correlations. In contrast, there was a poor concordance between either of these techniques and non-quantitative MSP. For biological samples with scarce DNA, QM-MSP is the method of choice.

The Use of Capital Budgeting Techniques in Businesses: A Perspective from the Western Cape

Capital budgeting is one of the most important areas of financial management. There are several techniques commonly used to evaluate capital budgeting projects namely the payback period, accounting rate of return, present value and internal rate of return and profitability index. Recent studies highlight that financial managers worldwide favor methods such as the internal rate of return (IRR) or non-discounted payback period (PP) models over the net present value (NPV), which is the model academics consider superior. In particular this research focused on small, medium and large businesses and investigated a number of variables and associations relating to capital budgeting practices in businesses in the Western Cape province of South Africa. The results revealed that payback period, followed by net present value, appears to be the most used method across the different sizes and sectors of business. It was also found that 64% of businesses surveyed used only one technique, while 32% of the respondents used between two to three different types of techniques to evaluate capital budgeting decisions. The findings show that the more complicated methods such as IRR and NPV are most favored by the large businesses as compared to the small businesses. The majority of the respondents believed that project definition was the most important stage in the capital budgeting process. Implementation stage appeared to be the most difficult stage for the manufacturing sector whereas Project definition, Analysis and selection and Implementation were generally rated as being the difficult stages by the retail sector. Project definition and Analysis and selection were found to be the most difficult stages by the service sector. Most businesses used the cost of bank loan as a basis in capital budgeting and more than two thirds of respondents used non-quantitative techniques to consider risk when making a decision on investing in fixed assets.

The Role of Pyrosequencing in Head and Neck Cancer Epigenetics

To evaluate promoter methylation quantitation using recently described pyrosequencing techniques by correlation with messenger RNA (mRNA) expression. DNA was extracted from tissue samples and was subjected to bisulphite conversion. Quantitative methylation data for multiple CpG sites in each of 9 gene promoters were obtained for tumors using pyrosequencing. RNA was extracted and converted to complementary DNA, and this formed the template for relative quantitation assays of the expression of each gene by real-time reverse transcription-polymerase chain reaction. Academic research. Thirty-seven patients with head and neck squamous cell carcinoma. The genes studied were P16 (OMIM 600160), cyclin A1 (OMIM 604036), RARB (OMIM 180220), E-cadherin (OMIM 192090), MGMT (OMIM 156569), STAT1 (OMIM 600555), ATM (OMIM 607585), hMLH1 (OMIM 120436), and TIMP3 (OMIM 188826). Immunohistochemistry was also performed for p16. STAT1, TIMP3, ATM, and hMLH1 promoters were essentially unmethylated in all cases. The data for cyclin A1 (Spearman rank correlation, rho = -0.53; P < .001), MGMT (rho = -0.53, P < .001), and RARB (rho = -0.34, P =.02) showed the expected negative correlation between levels of methylation and mRNA expression. The data relating to E-cadherin were inconclusive. Surprisingly, P16 expression was statistically significantly greater in those cases with higher levels of methylation (rho = 0.57, P < .001), a finding at odds with assumptions usually made in the literature relating gene promoter methylation to reduced gene expression. The results from p16 immunohistochemistry were in keeping with the mRNA data, but the number of positive staining samples proved too few for statistical analysis. These data present a novel perspective on head and neck cancer epigenetics and reveal new and some unexpected associations and findings. The advantages of pyrosequencing over nonquantitative techniques are discussed in analyses of this nature.

A Study of the Symbolic Aspects of Space Using Nonquantitative Techniques of Analysis

This article has two aims: first, to define and analyse the theoretical implications of the concepts of social urban identity and symbolic urban space (Valera, 1993, 1996, 1997; Valera and Pol, 1994) from within the integrated perspective provided by Environmental Psychology and Social Psychology; and second, to present an empirical study of these concepts based on nonquantitative data collection and analysis. Texts recorded in discussion groups were studied using Contextual Content Analysis (McTavish and Pirro, 1990) to determine the social identity of the inhabitants of a neighbourhood within the city of Barcelona and the meaning given to those spaces which they consider to be symbolic or representative of the neighbourhood. Finally, the results, as well as the advantages of this type of analysis, which are rarely used in Environmental Psychology, are discussed.

Sites of transcription of the Müllerian inhibiting substance gene in mouse testis

We have examined the transcription of Müllerian inhibiting substance (MIS) in testis by the sensitive technique of reverse transcription polymerase chain reaction (RTPCR). A developmental study of testis by this nonquantitative technique showed expression at all postnatal stages, including adults while liver and kidney provided negative controls. Cell separation studies indicated that highly purified interstitial cells, as well as less homogeneous Sertoli cell-enriched and germ cell-enriched fractions, contained RNA for MIS. The transcription of MIS in an interstitial cell type was confirmed by finding MIS mRNA in purified Leydig cells. Inasmuch as the germ cell-enriched fraction contains some Sertoli cells, and XX,Sxra and XX,Sxrb which have germ cell-depleted testes, contain MIS mRNA, a Sertoli cell source remains likely for the seminiferous tubule compartment.

Physician supply strategy: the case of the south.

The year 1989 marked the fourteenth successive year of steep decline in the number of U.S. medical school applicants. Many deans were beginning to think the unthinkable–that they might soon be unable to fill their entering classes with qualified students. In that year, the Eisenhower Center for the Conservation of Human Resources at Columbia University proposed to The Pew Charitable Trusts a study of the shrinking applicant pool as it affected eleven southern states: Alabama, Arkansas, Florida, Georgia, Kentucky, Louisiana, Mississippi, North Carolina, South Carolina, Tennessec, and Virginia. Support was granted by Pew for the period 1989–1991, and work got under way in the fall of 1989. Using a methodology that combined quantitative and nonquantitative techniques, the study explored the implications of a continuing dearth of well-qualified medical school applicants, schools’ responses to the challenge, and the longerterm measures that state legislatures might consider to ensure essential health services for all, even with fewer new physicians. Although applications to medical schools were down nationwide, the decision to focus the investigation on the South offered several advantages: the ability to differentiate regional from national factors; the presence of a heterogeneous medical education system with a large number of new (post1960) schools, and a below-average physician-to-population ratio. An increase in the number of medical school aspirants was reported in 1989 and has continued; nevertheless, the pool remains well below its peak, and problems persist. In this report, we summarize the experiencc of the South, which parallels that of

Strategic Planning in the US Pharmaceutical Industry: A National Study

The purpose of this study was to investigate the current state of strategic planning in the United States' pharmaceutical industry. Information specific to the strategic planning process in the industry was collected and analyzed. Of the original 142 companies solicited, 52 companies responded. The respondents represented both generic and brand name firms. Drug firms used a wide variety of techniques in their strategic planning processes. Non-quantitative techniques were found to be favored over quantitative methods. The actual strategic planning process was found to be consistent throughout the industry, although strategic planning was carried out to a greater extent by brand name pharmaceutical companies. Further analysis revealed the size of pharmaceutical companies displayed a strong positive correlation to strategic planning. Additionally, a significant positive correlation between industry sector and formal planning was identified. A statistically significant difference was noted in planners' opinio...

The beta 4 subunit of the integrin family is displayed on a restricted subset of endothelium in mice.

More than 15 subunits of the integrin family of cell surface adhesion molecules have been identified. The alpha 6 beta 4 integrin has recently been identified as a component of hemidesmosomes of stratified squamous epithelium. The monoclonal antibody (mAb) 346-11A binds to the beta 4 subunit in mice. Sequence analysis of a cDNA clone coding for this epitope localizes reaction of the mAb to a portion about half way through the extracellular domain at the beginning of the cysteine-rich region. Sites of beta 4 expression in mice were detected by autoradiographic analysis of tissues collected from mice 24 or 48 h after intravenous injection of 125I-labeled mAb 346-11A. This non-quantitative technique emphasizes detection of antigen exposed in the vascular space. These data show that in addition to the epithelia of several organs, the endothelia of intermediate vessels throughout the body are sites of beta 4 expression. In particular, endothelium of larger vessels but not capillaries in lung, vessels in thymus, spleen and Peyer's patches, and portal vessels but not the central veins of the liver, are positive. The expression of the beta 4 integrin in blood vessels may indicate a specialized function for beta 4 at these sites that is distinct from its role in hemidesmosome-mediated attachment.

Osteomyelitis and soft-tissue infection: differential diagnosis with 24 hour/4 hour ratio of Tc-99m MDP uptake.

The lesion-to-nonlesion 24 hour/4 hour ratio of technetium-99m methylene diphosphonate (MDP) uptake was used to distinguish osteomyelitis from increased bone uptake caused by adjacent soft-tissue infection. In a prospective study, this ratio was measured in 38 patients with 41 sites of increased uptake that were suspected to be osteomyelitis. The mean ratio was 1.18 +/- 0.18 in patients with proved osteomyelitis, which was significantly higher (P less than .001) than that in patients with increased uptake resulting from soft-tissue infection (0.98 +/- 0.05). On the basis of a receiver operating characteristic curve, a ratio of 1.06 was selected as the cutoff for differentiating osteomyelitis from soft-tissue infection. This method resulted in a sensitivity of 82%, a specificity of 92%, and an accuracy of 85%, and it appears to be more reliable than three- and four-phase scintigraphy, which are subjective and nonquantitative techniques.

POLITICAL RESOLUTION OF ENVIRONMENTAL CONFLICTS1

ABSTRACT Many water resources management problems ultimately require a political decision in order to formulate a course of action to follow. Metagame analysis is a nonquantitative technique that can model the decision‐making process and is used in this article to arrive at a politically feasible solution to a typical water pollution problem. The output from the linear programming formulation of the problem is used as input to the “political” metagame analysis. This allows a solution to the pollution conflict that is not only physically and economically feasible according to the linear programming analysis, but politically feasible as well.

How good are decision makers?

The job of a manager is to make decisions, but not everyone will agree that managers are good at their jobs. This article suggests that managers are not as good as one might expect; if they were, we would not have as evidence the condition of Penn Central Railway, Lockheed Aircraft, Pan Am, and other troubled corporations. The difficulties of decision making and of evaluating the results of decisions are worsened by arguments over quantitative and nonquantitative techniques, the shadow of ignorance which envelops all managerial decisions, and the lack of tools or techniques with which to measure the value or effectiveness of decisions once they are made. Other difficulties in evaluating the results of decisions arise from different perceptions of the purpose of the decision, time lapses, and occurrences over which the decision maker has no control. In fact, the overriding problem in evaluating managerial decisions is that there are no accepted measurements other than the judgment of the evaluator. Research has not been very rewarding in providing much needed measurement tools. Accordingly, this article offers no solutions to the formidable problem of finding measuring devices for decision makers but, tongue in cheek, offers an analogy wherein measurements are available.