Effects Of Tissue Type Research Articles

Trace Element Accumulation in Two Turtle Species, Malaclemys terrapin and Chelydra serpentina, in New Jersey, USA.

Trace elements in aquatic environments pose a risk to biological communities; this study investigates the total concentrations of arsenic (As), silver (Ag), cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), mercury (Hg), nickel (Ni), selenium (Se), lead (Pb), and zinc (Zn) within muscle, carapace, liver, and adipose tissues of diamondback terrapins and common snapping turtles in New Jersey. The effects of tissue type, sex, size, and location upon trace element accumulation were studied. The data obtained indicates that within diamondback terrapins and common snapping turtles, trace element accumulations displayed a significant difference among tissue types and sex (p < 0.005). The data indicates that Ag, Cd, Cu, and Hg can accumulate within the liver of diamondback terrapin. Se was found to accumulate in the livers of both diamondback terrapin and common snapping turtles. The highest mean concentrations of Co, Cr, Ni, and Pb were found in the carapace of both turtle species. Sex was found to have an impact on As, Hg, and Zn accumulation within different tissue types of diamondback terrapins. Diamondback terrapin males were found to have higher concentrations of As within the carapace. Diamondback terrapin females possessed higher concentrations of Hg in muscle tissues and Hg and Zn in the carapace. Turtle size and collection location land type and land cover did not display any correlation with trace element bioaccumulation for either species.

Mechanical properties of spinal cord grey matter and white matter in confined compression

To better understand the link between spinal cord impact and the resulting tissue damage, computational models are often used. These models typically simulate the spinal cord as a homogeneous and isotropic material. Recent research suggests that grey and white matter tissue differences and directional differences, i.e. anisotropy, are important to predict spinal cord damage. The objective of this research was to characterize the mechanical properties of spinal cord grey and white matter tissue in confined compression.Spinal cords (n = 12) were harvested immediately following euthanasia from Yorkshire and Yucatan pigs. The spinal cords were flash frozen (60 s at -80 °C) and prepared into four types of test samples: grey matter axial, grey matter transverse, white matter axial, and white matter transverse. Each sample type was thawed, and subsequently tested in confined compression within 6 h of euthanasia. Samples were compressed to 10% strain at a quasi-static strain rate (0.001/sec) and allowed to relax for 120 s. A quasi-linear viscoelastic model combining a first-order exponential with a 1-term Prony series characterized the loading and relaxation responses respectively. The effect of tissue type (grey matter vs. white matter), direction (axial vs. transverse), and their interaction were evaluated with a two-way ANOVA (p < 0.05) with peak stress, aggregate modulus, and relaxation time as dependent variables.This study found grey matter to be 1.6–2 times stiffer than white matter and both grey and white matter were isotropic in compression. These findings should be emphasized when studying SCI biomechanics using computational models.

Identification of housekeeping genes as references for quantitative real-time RT-PCR analysis in Misgurnus anguillicaudatus

Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) is a well-known method to quantify gene expression by comparing with the reference genes. Generally, housekeeping genes were set as references, as for their stable expression in varying conditions. Here, we try to evaluate few of such genes to identify suitable housekeeping genes as references for qRT-PCR analysis of gene expression in Misgurnus anguillicaudatus. This study evaluated the expression of four commonly used housekeeping genes, i.e. b-actin (ACTB), elongation factor 1 alpha (EF-1a), glyceraldehyde-3-phosphate (GAPDH) and 18S ribosomal RNA (18S rRNA), in gender difference, effects of tissue type, different developmental stages, chemical treatment of embryos/larvae with commonly used vehicles for administration and agents that represent known environmental toxicant. Rank ordering of expression stability was done using geNorm, NormFinder and BestKeeper algorithms. Results suggested that in the qRTPCR test, in all the experimental conditions, EF-1a could be selected as reference gene when analysing a target gene. For the study of different development stages, ACTB could be a candidate as reference gene. For the studies associated with different gender and tissue types, EF-1a would be better targeted as reference gene. Meanwhile, in toxicant treatment, expression of EF-1a seems to be more stable than others and could be considered as reference gene. This study could provide useful guidelines that can be expected to aid M. anguillicaudatus researchers in their initial choice of housekeeping genes for future studies and enable more accurate normalization of gene expression data.

Development and Application of a Multiplex Real-Time PCR Assay as an Indicator of Potential Allergenicity in Citrus Fruits.

The effects of tissue type, harvest maturity, and genetic factors on the expression of genes that related to citrus fruit allergies remain poorly understood. In the present study, a multiplex real-time PCR assay was developed to monitor the expression of citrus allergen genes individually with the advantages of much fewer sample requirements and simultaneously multiple target genes detection. Gene specific primer pairs and Taqman probes of three citrus allergen genes Cit s 1.01, Cit s 2.01, and Cit s 3.01 and the house-keeping gene β-actin were designed based on gene sequence differences. The PCR results showed that differential expression patterns were found during the ripening process. The expression levels of Cit s 3.01 were much higher than those of Cit s 1.01 and Cit s 2.01 in both peel and pulp tissues among 10 citrus cultivars. Data suggested that Kao Phuang Pummelo could be safely consumed with a potential low risk in allergenicity. Considering that assessing allergenicity is one of the tests in food safety, this assay might also facilitate the breeding and production of "allergy-friendly" citrus fruits.

Surface energy of bovine dentin and enamel by means of inverse gas chromatography.

Adhesion between tooth tissues and dental fillings depends on the surface energy of both connected materials. Bond strength can be determined directly or indirectly as a work of adhesion on the basis of values of surface energy of these materials. Inverse gas chromatography (IGC) is one of the methods of surface energy examination. In this study the values of total surface energy components of wet and dry teeth fragments (enamel, crown dentin and root dentin) were determined with the use of inverse gas chromatography. Inverse gas chromatography has never been used for investigation of surface energy of natural tooth tissues. Different storage conditions were examined - wet and dry. Different values of surface energy are observed according to the type of tooth tissue (dentin or enamel), occurring place (crown or root) and storage conditions (dry or wet). The effect of tissue type and occurring place was the greatest, while storage conditions were of secondary importance. Surface energy depends on composition of tissue, its surface area and the presence of pores.

Tissue cryobanking for conservation programs: effect of tissue type and storage time after death

In this study, we investigated the temporal post-mortem limits, within which there will be guarantees of obtaining living cells from several tissues of sheep and cattle and the effect of vitrification on the ability of cells from tissue stored at different times. Muscle tissue and auricular cartilage were stored at 4°C for 5, 48, 72, 96 and 216 h post-mortem (hpm). Tissue samples were sorted into two groups: one group was in vitro cultured immediately after storage and the other was vitrified after storage and then in vitro cultured. In cattle and sheep, no differences in subconfluence rates were observed between the two experimental groups. At the same time, no significant differences were observed in the number of days required in culture to reach confluence between non-vitrified and vitrified groups when tissues were stored at 4°C for different times. In sheep, while the population doubling times (PDT) were similar in cartilage cells from vitrified and non-vitrified tissues and stored at 4°C for 5 and 216 hpm, PDT of muscle cells were longer in 216 hpm stored groups than in 5 hpm stored groups. In bovine, although the PDT of muscle cells were similar for 5 and 216 hpm and both vitrified and non-vitrified tissues and the PDT were longer in cartilage cells from vitrified than from non-vitrified tissues. In conclusion, although storage times and vitrification have different effects on tissues from cattle and sheep, this study showed that living cells could be obtained from all groups. Therefore, cartilage and muscle tissues can be stored at 4°C for 216 hpm and used for cyrobanking.

Detection of acute fentanyl exposure in fresh and decomposed skeletal tissues part II: The effect of dose–death interval

The effects of dose–death interval on the detection of acute fentanyl exposure in fresh and decomposed skeletal tissues (marrow and bone), by automated enzyme-linked immunosorbent assay (ELISA) are described. Rats ( n = 14) were administered fentanyl acutely at a dose of 0 ( n = 2) or 60 μg/kg ( n = 12) by intraperitoneal injection, and euthanized within 20, 45, 135, or 225 min. Femora and tibiae were extracted from the fresh corpses and marrow was isolated from the femoral and tibial medullary cavities. The remains were then allowed to decompose outdoors to the point of complete skeletonization, and vertebrae, pelvi and miscellaneous (humeri and scapulae) were recovered for analysis. In all cases, bones were cleaned in alkaline solution and then ground into a fine powder. Marrow was homogenized in alkaline solution. Fentanyl was extracted from ground bone by methanolic extraction. Extracts were adjusted to pH 6 and analyzed by ELISA. Perimortem heart blood was also collected and diluted in phosphate buffer prior to screening by ELISA. The effect of tissue type on ELISA response was examined through determination of binary classification test sensitivity and the relative decrease in absorbance (%DA, drug-positive tissues vs. drug-free controls) in each tissue type. Overall, the %DA varied significantly between extracts from different skeletal tissues at a given dose–death interval, according to the general order of marrow > decomposed bone > fresh bone. Binary classification test sensitivity values for fentanyl in marrow, fresh epiphyseal (femoral and tibial) bone, fresh diaphyseal (femoral and tibial) bone, decomposed vertebrae, decomposed pelvic bone, and decomposed miscellaneous bone were 67–100%, 0–33%, 0–33%, 0–67%, 0–67% and 0–33%, respectively, over all dose–death intervals. Although group mean %DA values showed a strong negative correlation with dose–death interval in marrow, fresh epiphyseal bone, decomposed vertebrae, pelvic and miscellaneous bone ( r = −0.989, −0.930, −0.955, −0.903, and −0.974, respectively), the high variability in both fresh and decomposed bone precluded differentiation of the dose–death intervals based on %DA value alone. Overall, the results suggested that the type of skeletal tissue sampled may not be as important as the amount of residual marrow remaining in skeletonized remains.

Detection of acute fentanyl exposure in fresh and decomposed skeletal tissues

The detection of acute fentanyl exposure in fresh and decomposed skeletal tissues (marrow and bone), by automated enzyme-linked immunosorbent assay (ELISA) is described. Rats (n=15) were administered fentanyl acutely at a dose of 0 (n=3), 15 (n=3), 30 (n=3) or 60 microg/kg (n=6) by intraperitoneal injection, and euthanized within 20 min. Femora and tibiae were extracted from the fresh corpses and marrow was isolated from the femoral and tibial medullary cavities. The remains were then allowed to decompose outdoors to the point of complete skeletonization, and vertebrae and pelvi were recovered for analysis. In all cases, bones were cleaned in alkaline solution and then ground into a fine powder. Marrow was homogenized in alkaline solution. Fentanyl was extracted from ground bone by methanolic extraction. Extracts were adjusted to pH 6 and analyzed by ELISA. Perimortem heart blood was also collected and diluted in phosphate buffer prior to screening by ELISA. The effect of tissue type on ELISA response was examined through determination of binary classification test sensitivity and the relative decrease in absorbance (%DA, drug-positive tissues vs drug-free controls) in each tissue type. Overall, the %DA varied significantly between extracts from different skeletal tissues under a given dose condition, according to the general order of marrow>vertebrae approximately pelvi>epiphyseal bone approximately diaphyseal bone. Binary classification test sensitivity values for fentanyl in marrow, fresh epiphyseal (femoral and tibial) bone, fresh diaphyseal (femoral and tibial) bone, decomposed vertebrae and decomposed pelvic bone were 100%, 16-33%, 0-16%, 0-33% and 66-100%, respectively, at the 60 microg/kg dose level. While mean %DA values showed a strong positive correlation with those in marrow and blood measurements and the administered dose (r=0.997 and 0.986), such a correlation was not observed in assays of decomposed tissues (r=-0.157 and -0.315). These results suggest that the type of skeletal tissue sampled and position within a given bone may be important considerations in the choice of substrate for fentanyl screening in skeletal tissues, but that quantitative analysis of drugs in decomposed bones may be of limited interpretive value.

Utility of Immunoassay in Drug Screening in Skeletal Tissues: Sampling Considerations in Detection of Ketamine Exposure in Femoral Bone and Bone Marrow Following Acute Administration Using ELISA*

Detection of ketamine exposure in skeletal tissues by automated enzyme-linked immunosorbent assay (ELISA) and gas chromatography with electron capture detection (GC-ECD) is described. Rats (n = 18) received 0, 15, 30, or 75 mg/kg ketamine hydrochloride acutely (i.p.), and were euthanized within 15 min or 1 h. Ketamine was extracted from ground femoral bone by methanolic incubation followed by liquid-liquid extraction (LLE), while marrow was homogenized in alkaline solution, and then underwent LLE. Extracts were analyzed by ELISA, and subsequently by GC-ECD following derivatization with trifluoroacetic acid anhydride. The effect of tissue type (i.e., diaphyseal bone vs. epiphyseal bone vs. bone marrow) on the immunoassay response was examined through determination of binary classification test sensitivity (S) and measurement of the relative decrease in absorbance (%DA, drug-positive tissues vs. drug-free controls) in each tissue type. The %DA varied significantly between different tissues examined under a given dose condition, and generally decreased in the order marrow > epiphyseal bone > diaphyseal bone, at all dose levels examined. Measured S values for marrow, epiphyseal bone, and diaphyseal bone were 100%, 77%, and 23%, respectively (75 mg/kg dose). These results suggest that the type of skeletal tissue sampled and position sampled within a given bone (diaphyses vs. epiphyses) are important parameters in drug screening of skeletal tissues.

Effects of Tissue Type and the Dose-Death Interval on the Detection of Acute Ketamine Exposure in Bone and Marrow with Solid-Phase Extraction and ELISA with Liquid Chromatography-Tandem Mass Spectrometry Confirmation

Ketamine exposure was detected in skeletal tissues by ELISA and liquid chromatography-tandem mass spectrometry (LC-MS-MS). Rats (n = 9) received ketamine hydrochloride acutely (75 mg/kg, i.p.) and were euthanized within 15, 30, or 90 min. Drug-free control animals (n = 3) were also euthanized. Extracted femora were separated into epiphyseal and diaphyseal fragments, with marrow isolated from the medullary cavity. Bone was ground and incubated in methanol. Extracts were dried and reconstituted in phosphate buffer (0.1 M, pH 7.3), and marrow was homogenized in alkaline solution. Both then underwent solid-phase extraction. Extracts were assayed by ELISA, with data expressed in terms of relative decrease in absorbance (%DA, drug-positive tissues vs. matrix-matched drug-free controls) and binary classification test sensitivity (S). Generally, %DA decreased in the order of marrow > epiphyseal bone > diaphyseal bone, and was negatively correlated with dose-death interval (DDI). Measured S values were 100% in ELISA analysis of extracts of all tissue types. Sensitivity values were computed from LC-MS-MS data using a 5 ng/mL cutoff. Sensitivity values for ketamine detection were 100%, 0-100% and 0%, at the 15, 30, and 90 min DDI, respectively, and sensitivity values for norketamine detection were 0-66%, 0-66%, and 0% at the 15, 30, and 90 min DDI, respectively. These results suggest that the tissue type sampled and DDI may influence the sensitivity of detection of ketamine exposure in skeletal tissues.

Characterization of housekeeping genes in zebrafish: male-female differences and effects of tissue type, developmental stage and chemical treatment

BackgroundResearch using the zebrafish model has experienced a rapid growth in recent years. Although real-time reverse transcription PCR (QPCR), normalized to an internal reference ("housekeeping") gene, is a frequently used method for quantifying gene expression changes in zebrafish, many commonly used housekeeping genes are known to vary with experimental conditions. To identify housekeeping genes that are stably expressed under different experimental conditions, and thus suitable as normalizers for QPCR in zebrafish, the present study evaluated the expression of eight commonly used housekeeping genes as a function of stage and hormone/toxicant exposure during development, and by tissue type and sex in adult fish.ResultsQPCR analysis was used to quantify mRNA levels of bactin1, tubulin alpha 1(tuba1), glyceraldehyde-3-phosphate dehydrogenase (gapdh), glucose-6-phosphate dehydrogenase (g6pd), TATA-box binding protein (tbp), beta-2-microglobulin (b2m), elongation factor 1 alpha (elfa), and 18s ribosomal RNA (18s) during development (2 – 120 hr postfertilization, hpf); in different tissue types (brain, eye, liver, heart, muscle, gonads) of adult males and females; and after treatment of embryos/larvae (24 – 96 hpf) with commonly used vehicles for administration and agents that represent known environmental endocrine disruptors. All genes were found to have some degree of variability under the conditions tested here. Rank ordering of expression stability using geNorm analysis identified 18s, b2m, and elfa as most stable during development and across tissue types, while gapdh, tuba1, and tpb were the most variable. Following chemical treatment, tuba1, bactin1, and elfa were the most stably expressed whereas tbp, 18s, and b2m were the least stable. Data also revealed sex differences that are gene- and tissue-specific, and treatment effects that are gene-, vehicle- and ligand-specific. When the accuracy of QPCR analysis was tested using different reference genes to measure suppression of cyp19a1b by an estrogen receptor antagonist and induction of cyp1a by an arylhydrocarbon receptor agonist, the direction and magnitude of effects with stable and unstable genes differed.ConclusionThis study provides data that can be expected to aid zebrafish researchers in their initial choice of housekeeping genes for future studies, but underlines the importance of further validating housekeeping genes for each new experimental paradigm and fish species.

Quantitative characterization of hemodynamic properties and vasculature dysfunction of high‐grade gliomas

Aberrations in tumor and peritumoral vasculature may not be distinguishable by cerebral blood flow (CBF) or cerebral blood volume (CBV) alone. The relationships between CBF and CBV were examined to estimate vasculature-specific hemodynamic characteristics. Twenty glioma patients were studied with dynamic susceptibility T2*-weighted MRI [(dynamic contrast-enhanced magnetic resonance imaging (DSC-MRI)] before and during week 1 and 3 of radiotherapy (RT). CBF and CBV were calculated from DSC-MRI, and relationships between the two were evaluated: the physiological measure of mean transit time (MTT) = CBV/CBF; empirical fitting using the power law CBV = constant x (CBF)(beta). Three different tissue types were assessed: the Gd-enhancing tumor volume (GEV); non-enhanced abnormal tissue located beyond GEV but within the abnormal hyperintense region on FLAIR images (NEV); normal tissue in the hemisphere contralateral to the tumor (CNT). The effects of tissue types, CBV magnitudes (low, medium and high), before and during RT, on MTT and beta were analyzed by analysis of variance (ANOVA). The MTT and beta for the three tissue types were significantly different (p < 0.009). MTT increased from CNT (1.60 s) to NEV (1.93 s) to GEV (2.28 s) (p < 0.0005). beta was significantly greater in GEV (1.079) and NEV (1.070) than in CNT (1.025). Beta increased with increasing CBV magnitude while MTT was independent of CBV magnitude. There was a significant decrease in MTT of NEV and GEV during week 3 of RT compared with pre-RT values for all CBV magnitudes. There was a significant increase in beta during RT in the tumor and peritumor. Progressive abnormalities in vasculature and hemodynamic characteristics of the vascular bed were delineated, with significant disorder in the tumor but mild abnormality in peritumoral tissue.

TH‐E‐ValB‐06: Quantitative Characterization of Tumor Vascular Dysfunction in High‐Grade Gliomas Prior to and During Radiotherapy

Introduction: Vascular properties within and adjacent to tumors may not be distinguishable by cerebral blood flow [CBF] or cerebral blood volume [CBV] alone, since the rates of CBV change may not be proportional in magnitude to CBF change. Hence, the empirical and physiological relationships between CBF and CBV were examined to estimate vasculature‐specific hemodynamic characteristics in high‐grade gliomas. Methods: Twenty patients with gliomas were studied with dynamic contrast‐enhanced T2* MRI [DCE‐MRI] before and during radiotherapy [RT]. CBF and CBV were calculated from DCE‐MRI and the relationships between the two were evaluated using two different metrics: The physiological measure of Mean Transit Time [MTT]=CBV/CBF; and, Empirical fitting of CBV and CBF using the power law, expressed as CBV=Constant*(CBF)β. Three tissue types were assessed, Gd‐enhanced tumor volume [GdTV], non‐enhancing abnormal tissue located beyond GdTV but within the abnormal hyperintense volume on FLAIR images [NEV], and normal tissue in hemisphere contralateral to tumor [CNT]. Effects of tissue types, CBV magnitudes (low[L], medium[M] and high[H] CBV), before and during RT, on MTT and β were analyzed by factorial ANOVA. Results: Both, MTT and β were significantly different (p&lt;0.009) among the three tissue types. MTT increased from CNT(=1.60s) to NEV(=1.93s) to GdTV(=2.28s) (&lt;0.0005). The power exponent β was significantly greater in GdTV(=1.079) and NEV(=1.070) than CNT(=1.025), but β in NEV and GdTV were not significantly different from each other. β increased with increasing CBV magnitude. There was a significant decrease in MTT and a significant increase in β in tumor (GdTV) and peritumoral (NEV) tissue during RT compared with pre‐RT values. Conclusions: β was strongly dependent on CBV magnitude and MTT on tissue type. Progressive abnormalities in functional characteristics of the vascular bed were noted, with significant disorder in tumor, but mild abnormality in peritumoral tissue. Early vascular response to radiation was first observed in functional rather than structural properties.

Effects of Tissue-type and Development on Dark Respiration in Two Herbaceous Perennials

Perennial plants go through a number of developmental stages during the growing season. Changes in metabolism during these phases have been documented in laboratory-grown plants but never in native plants growing in natural habitats. The purpose of this study was to describe the seasonal pattern of dark respiration in the above-ground tissues of two herbaceous perennials, Bistorta bistortoides(Pursh) Small and Campanula rotundifolia L., growing in the Rocky Mountains (USA). The effect of biomass accumulation on respiration rate and differences in respiration rate among tissues were measured. Respiration rate differed significantly among the above-ground tissues. Reproductive structures had the highest respiration rates, followed by leaves, then stems. Respiration rate decreased by 10–90% over the growing season in these tissues but was generally not correlated with a decrease in biomass accumulation. The seasonal pattern of respiration rate varied significantly among tissues. Total tissue respiratory flux was calculated at 15°C for each tissue. In both species, total above-ground respiratory flux was either relatively constant during the growing season with a marked decrease at seed dispersal or a maximum rate was reached at mid-season. In B. bistortoides, leaves had the highest total respiratory fluxes, and the respiratory fluxes of the stem and reproductive structures were similar to one another. In C. rotundifolia, leaf and stem respiratory fluxes were similar, while the respiratory flux of the reproductive structures was considerably lower than that of leaves and stems. This study emphasizes the importance of developmental processes and tissue-type on respiration rate and highlights the importance of including all plant tissues in predictive models of plant carbon balance.

Oxytocin receptor expression in human term and preterm gestational tissues prior to and following the onset of labour.

Oxytocin receptor (OTR) mRNA expression has previously been demonstrated in human myometrium, decidua, chorion and amnion but the effect of gestational age and the onset of labour has not been determined in these individual tissues. Spatial OTR mRNA expression was examined by in situ hybridization and ligand binding was confirmed using autoradiography with the iodinated oxytocin antagonist d(CH2)5[Tyr(Me)2,Thr4,Tyr-NH29]-vasotocin (125I-OTA). Tissue was collected at term (>37 weeks of gestation) or preterm (24-36 weeks of gestation) caesarean section and classified as labour (contractions every 5 min associated with cervical dilatation) or non-labour. OTR mRNA expression was measured as optical density units from autoradiographs. There was a highly significant (P<0.001) effect of tissue type on expression of OTR mRNA with expression greatest in myometrium, low in decidua and chorion and not detected in placenta. Similar results were obtained with the 125I-OTA-binding studies, indicating that the message was translated. Amnion had an apparently high level of both hybridization and 125I-OTA binding in some samples, but a lack of specificity prevented quantification of the signal in this tissue type. Term myometrium (labour and non-labour) had significantly higher (P<0.01) OTR mRNA expression than preterm myometrium, but there was no further increase in mRNA concentration associated with labour onset. In contrast, 125I-OTA binding in myometrium was already high at 33 weeks and did not increase further either later in pregnancy or with labour. In decidua there was no effect of gestational age or labour onset on OTR mRNA expression or 125I-OTA binding. In summary, OTR mRNA expression in the myometrium increased in late pregnancy whereas decidual expression was much lower and did not rise at term.

Performance estimation of diagnostic tests for cervical precancer based on fluorescence spectroscopy: effects of tissue type, sample size, population, and signal-to-noise ratio.

Fluorescence spectroscopy may provide a cost-effective tool to improve precancer detection. We describe a method to estimate the diagnostic performance of classifiers based on optical spectra, and to explore the sensitivity of these estimations to factors affecting spectrometer cost. Fluorescence spectra were obtained at three excitation wavelengths in 92 patients with an abnormal Papanicolaou smear and 51 patients with no history of an abnormal smear. Bayesian classification rules were developed and evaluated at multiple misclassification costs. We explored the sensitivity of classifier performance to variations in tissue type, sample size, tested population, signal to noise ratio (SNR), and number of excitation and emission wavelengths. Sensitivity and specificity could be evaluated within +/- 7%. Minimal decrease in diagnostic performance is observed as SNR is reduced to 15, the number of excitation-emission wavelength combinations is reduced to 15 or the number of excitation wavelengths is reduced to one. Diagnostic performance is compromised when ultraviolet excitation is not included. Significant spectrometer cost reduction is possible without compromising diagnostic ability. Decision-analytic methods can be used to rate designs based on incremental cost-effectiveness.

Effect of tissue type and variety on cell wall chemistry of onion ( Allium cepa L.)

The purpose of this study was a comparative examination of the cell wall chemistry of component tissues of different varieties of onions ( Allium cepa L. cv Sturon, Durco, Hysam, Grano de Oro and Caribo). Cold alcohol-insoluble residues (CAIRs) were prepared and were extracted sequentially with water, imidazole, CDTA, Na 2CO 3 and 0.5 M KOH to leave a residue. These were analysed for their carbohydrate compositions. On a whole organ basis, the cell wall carbohydrate composition was similar for each variety studied, and no significant change resulted from commercial storage, with or without sprout suppressant. However, there were significant differences in the carbohydrate composition of cell walls from different tissues. Cell walls of inner leaf bases contained galactoserich pectic polysaccharides. Outer layers had progressively less pectic galactose, and the outer brown skin contained virtually none. This was accompanied by changes in water- and CDTA-soluble polysaccharides and may be related to the dry and protective nature of the non-lignified outer skin. The differences in cell walls from different tissues provide the basis for developing processing methods for exploiting onion waste.

Effects of tissue type and promoter strength on transient GUS expression in sugarcane following particle bombardment

Effects of tissue type and promoter strength on transient GUS expression in the sugarcane (Saccharum spp. hybrids) cultivar NCo 310 were evaluated following microprojectile bombardment of leaf explants. GUS expression was histochemically or fluorometrically measured 48 h after delivery of the uidA gene. High levels of GUS expression were obtained in leaf segments isolated from young, expanding sugarcane leaves cultured for 1, 3, or 6 d prior to bombardment. The promoter derived from the maize ubiquitin 1 gene (Ubi-1) produced significantly more GUS foci and higher GUS activity levels compared to the recombinant Emu, rice actin 1 (Act1), and CaMV 35S promoters. Our transient expression system should facilitate efforts to identify promoters and elements which will regulate desired gene expression patterns in sugarcane and aid in development of an efficient stable transformation system.

Identification of Raspberry Cultivars in Vivo and in Vitro Using Isoenzyme Analysis

Abstract Isoenzyme analysis of leaf tissue was used to characterize and separate 18 greenhouse-grown red raspberry (Rubus idaeus L.) cultivars and two purple raspberry (R. ×neglectus Peck) cultivars. Five enzymes were found to be useful: Phosphoglu-coisomerase (PGI), phosphoglucomutase (PGM), malate dehydrogenase (MDH), triose phosphate isomerase (TPI), and isocitrate dehydrogenase (IDH). Fourteen of the 20 cultivars were uniquely separated using these enzymes. The remaining six were assigned to three different groups. Staining for additional isoenzymes may lead to the separation of cultivars within these groups. The effect of tissue type on isoenzyme banding patterns was determined by comparing different tissues from three cultivars. No differences were noted, except in the case of the enzyme PGM, for which differences in the relative densities of the bands were seen. For PGM, the best patterns were obtained from mature tissues (leaves, stems, or petioles). Three cultivars were micro-propagated from shoot tips in vitro and were subsequently analyzed by isoenzyme staining. The culture environment was not found to have any effect on isoenzyme phenotype. This result suggests that isoenzyme analysis can be used to identify raspberry cultivars grown in vitro.

Pulsed CO2 laser tissue ablation: Effect of tissue type and pulse duration on thermal damage

Tissue removal by infrared lasers is accompanied by thermal damage to nonablated tissue. The extent of thermal damage can be controlled by a choice of laser wavelength, irradiance, and exposure duration. The effect of exposure duration has been studied in vivo by using CO2 lasers with pulse widths that vary from 2 microseconds to 50 msec. Pulse widths of 50 msec, typical of a shuttered, continuous-wave CO2 laser, produce damage regions 750 micron wide in normal guinea pig skin; the use of a 2-microseconds-long pulse reduced this damage zone to as little as 50 micron. Using 2-microseconds-long pulses, in vitro studies showed that the minimum zone of thermal damage varied significantly with tissue type. The thermal denaturation of these tissues has been studied and correlated with damage. The effect of denaturation temperature and pulse duration on the width of the damage zone is explained by a simple model.