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Kindle Money Mastery

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Vascular defects in different disorders of pregnancy

Findings overlap with observations in ''classical'' pre-eclampsia (Aardema et al., 2001). In our own series, 7 out of 10 HELLP cases showed acute atherosis, which is a much higher incidence than observed in pre-eclampsia, and also the incidence of atherosis in myometrial segments (5 out of 10 cases) was unexpectedly high, compared with 8 in the pre-eclamptic group (unpublished results). These observations are compatible with the concept that the HELLP syndrome forms part of severe pre-eclampsia. It could be speculated that extension of this lesion over a longer distance of a vessel may compromise vascular function even further, also leading to an increased release of cytotoxic cytokines into the blood and surrounding tissues.

Therapeutic Goals

Johnson and Saliba first developed the material on body position presented here. They observed that more effective control of the patient's motion came when the therapist was in the line of the desired motion. As the therapist shifted position, the direction of the resistance changed and the patient's movement changed with it. From this knowledge they developed the following guidelines for the therapist's body position (G. Johnson and V. Saliba, unpublished handout 1985) The therapist's body should be in line with the desired motion or force. To line up properly, the therapist's shoulders and pelvis face the direction of the motion. The arms and hands al

The Uniform Thin Film Approximation UTFA

4 One way in which the assumptions of the UTFA could be invalidated even though minute nanoparticles are being adsorbed as a monolayer is if those particles cluster together to create large islands. Such behaviour is evidenced by a broadening of the TM incoupling peak but not the TE one (J.J. Ramsden and T. Jaworek, unpublished observations), and has a definite kinetic signature as discussed in Sect. 2.6.1.

Implications for the Production of Insect Resistant Transgenic Plants

Response of genes encoding serine proteases to dietary PI in larvae of the lepidopteran, Helicoverpa armigera (Bown et al, unpublished). mRNA levels for four different serine protease genes were assessed as a proportion of total RNA by Northern blotting of total RNA isolated from larval guts, followed by densitometry of the autoradiographs. Larvae were taken at the start of the fourth instar, and exposed to diet containing soybean Kunitz trypsin inhibitor (SKTI) at 2 of total protein, or control diet, for the stated times before RNA extraction. The differential expression of different protease mRNAs is shown by the strong up-regulation of sequences detected by cDNAs SR28 (trypsin-like) and SR42 (chymotrypsin-like) in the presence of SKTI, whereas sequences detected by cDNA SR40 (trypsin-like) are little affected, and sequences detected by HaTC16 (trypsin-like) are down-regulated relative to the control. Fig. 2.5. Response of genes encoding serine proteases to dietary PI in...

Expectations of Context at Intention Formation

In unpublished work from our laboratory we have also tested the effect of implying that cues would appear in a specific context (Marsh, Hicks, & Cook, 2002). In this case, we had three different conditions that were each given four specific animal exemplars to study at intention formation. In one case it was four specific words, in another condition it was four pictures, and in the third it was four animal words along with their corresponding pictures (i.e., both). We asked participants to form an intention to say the name of each out loud any time they encountered those four specific cues. The ongoing activity was to rate the frequency of occurrence in the past 2 weeks of a series of pictures and words intermixed with one another. Like Nowinski and Dismukes (2005), our instructions were clear and unambiguous that participants should respond regardless of the format (picture or word), but the page we handed them with the four cues implied that cues would occur in one or the other of...

Forest Tree Population Structures And The Adoption Of Cloning

A recent unpublished study in Finland has shown that natural aspen stands may consist of between 2 and 10 clones. It suggests that clonal aspen or hybrid aspen plantations could be arranged in a chessboard structure, each clone grown in pure monoculture of say 1 4 ha or about 500 ramets of a selected ortet. A 1-ha plantation would consist of 4 clones and in larger plantations, clone numbers could increase to 10 and the same clone could be repeated in another location of the large plantation area. Trees that do not form clones under natural conditions, such as conifers, should, however, be grown as clonal mixtures, thus imitating natural stands with their genetic variation between individuals.

Function of SR Genes in Senescence

Similar to leaf senescence, protein degradation has been demonstrated to be a major part of petal senescence and the remobilization of N to the developing ovary (Nichols, 1976). A few of the SR cysteine proteases have been shown to be up regulated in both leaves and petals (SAG 12, Quirino etal., 1999 DCCP1, Jones etal, 1995, Jones, unpublished Peth1, Tournaire et al., 1996). Large increases in proteolytic activity during the senescence of the ephemeral flower, daylily, have been well documented and recently this proteolytic activity was correlated with increases in the expression of two cysteine protease genes (Sen11 and Sen102) during the senescence of tepals (Valpuesta et al., 1995 Guerrero et al., 1998 Stephenson and Rubinstein, 1998). In contrast to the cysteine proteases from carnation and petunia, transcripts are not detectable in young daylily flowers (buds) and the level of transcript does not increase in senescing leaves (Guerrero et al., 1998). Both daylily cysteine...

Major Issues And Future Directions

Experiments are needed along these lines to ascertain whether our anecdote about returning to the room where one formed an intention indeed increases the probability of recollection. Further experiments are needed that go beyond the environmental context at encoding. There are a variety of internal contexts that, whether reinstated or not during a performance interval, may affect intention completion. One's mood or prevailing emotional state has not been investigated in any detail (but see Harris & Menzies, 1999). In unpublished work, we have tested obsessive-compulsive individuals and discovered that with an intention about neutral material they are disadvantaged in cue detection, but with an intention related to bodily fluids (e.g., mucus and urine) they are not. The idea there is that individuals with obsessive-compulsive disorder behave as cognitively distracted for material other than that related to their compulsion. Work along these lines...

ATPDependent Proteolysis in Archaea

As a 650 kDa complex with ATPase activity. Surprisingly, it hydrolyzes CTP even faster than ATP, and GTP and UTP to a lesser extent. When the 650 kDa S4 complex was mixed with proteasomes from T. acidophilum, degradation of substrate proteins was stimulated up to 25-fold, hence the complex was named PAN, for proteasome activating nucleotidase.9b By contrast, the degradation of short peptide substrates by the proteasome was not stimulated by PAN or ATP. Presumably, the small peptides can readily enter this particle by simple diffusion and can be degraded by the active sites which can function even in the absence of ATP or PAN (in contrast to those of HslV). Therefore, PAN probably functions to unfold the polypeptides and facilitate or modulate its entry into the 20S particle. Like many molecular chaperones and regulators of proteolysis (e.g., ClpA, ClpX, HslU), PAN is a protein activated ATPase and ATP hydrolysis was stimulated up to 2 fold by proteins that were degraded in an...

Genetically Modified Mice

We have reported that the targeted expression of ODC to the heart enhances P-adrenergic agonist-induced cardiac hypertrophy (21). Transgenic mice were generated with cardiac-specific expression of a stable ODC protein using the a-myosin heavy chain promoter (MHC-ODC mice). Founder lines with a 30-fold overaccumula-tion of putrescine, a fourfold elevation in spermidine, a slight increase in spermine, and accumulation of large amounts of cadaverine compared with littermate controls were established. Despite the alterations in polyamines in the transgenic heart, early studies detected no significant hypertrophy based on ratio of heart weight to body weight, although ANF mRNA was elevated in transgenic ventricles (21). Subsequent studies after multiple backcrosses have revealed a moderate baseline hypertrophy, as measured by ratio of heart to body weight, myocyte cross-sectional area, and echocardiography (29). Stimulation of P-adrenergic signaling by isoproterenol resulted in much more...

Neuro ImmuneEndocrine Regulation of the HPA Axis

1997), and IL-11R (Auernhammer et al., 1999) has been demonstrated. In primary cultures derived from human fetal pituitaries of 16-31 wk of gestation, recombinant human IL-6, LIF, and OSM treatments induced ACTH secretion either alone or in synergistic action with CRH (Shimon et al., 1997). In murine corticotroph AtT-20 cells, LIF, OSM, and IL-11 induce POMC gene expression and ACTH secretion (Akita et al., 1995 Ray et al., 1996 Auernhammer et al., 1999). LIF-induced POMC gene expression and ACTH secretion involve tyrosine phosphorylation and activation of Jak2 (Bousquet, unpublished data), gp130 (Ray et al., 1996), STAT3 mutants (Ray et al., 1996, 1998), and STAT1 (Ray et al., 1996,

The role of the placenta

The current evidence is that circulating placental debris is likely to be an important part of the systemic inflammatory stimulus associated with both normal and pre-eclamptic pregnancies. We have shown that they are directly damaging to endothelium (Smarason et al., 1993) which is stimulated to release pro-inflammatory substances (von Dadelszen et al., 1999). Our preliminary evidence is that they are directly pro-inflammatory (Sacks et al., unpublished observations Branton etal., unpublished observations). It is possible that it is this circulating debris which is the danger signal of pregnancy to which the inflammatory system responds appropriately.

Intendedness of Pregnancy and Preterm Delivery

Having an unintended pregnancy is estimated to increase the odds of delivering an infant of low birth weight by about 1.2 to 1.8 (IOM, 1995). However, only three studies on intentions and preterm birth were available as of 1995, and of those three studies, two were unpublished. On the basis of the findings from these preliminary studies, IOM (1995) indicated that the increased risk of low birth weight because of an unintended pregnancy appeared to be related to preterm delivery rather than intrauterine growth restriction. Subsequently, at least one further study has been conducted with a sample of 922 African-American, low-income pregnant women recruited in four hospital-based prenatal clinics in Baltimore, Maryland (Orr et al., 2000). Women with unintended pregnancies were 1.82 times more likely to deliver their infants pre-term, after adjustment for clinical and behavioral factors associated with preterm delivery.

Regulatory Genes and Intracellular Signaling

In addition to the delayed-senescence phenotype of the mutants in age and dark-induced senescence, these mutations delayed the responses to several phytohormones such as ethylene, abscisic acid (ABA), and methyl jasmonate (MeJA) (Woo, Park, and Nam, unpublished data). This suggests that ORE1, ORE3, and ORE9 may function at a common step of senescence affected by these factors. The ORE1, ORE3, and ORE9 genes may be required for proper progression of leaf senescence induced by the phytohormones, as well as age and darkness (Fig. 5-1). In fact, ore3, also designated as ein2-34, is an allele of ein2 (ethylene-insensitive2) mutations that are known to affect seedling responses to various hormones (see Section IVC). In contrast, the other mutants do not have a defect in general hormone responses but in hormone-induced senescence.

Evidence for increased placental free radical synthesis in preeclampsia

And cytotrophoblast (Dechend et al., 2003). Dechend et al. reported increased immunostaining for the NAD(P)H oxidase subunits in pre-eclamptic placentas (Dechend et al., 2003). Recently, we have reported similar NAD(P)H oxidase-mediated O2* generation in placental tissue of women with pre-eclampsia when compared to normotensive controls and women with early-onset pre-eclampsia showed higher placental O2* generation. The authors' unpublished preliminary data show that NAD(P)H oxidase is functionally active as early as the first trimester of pregnancy, raising the possibility that NAD(P)H oxidase could play a role in early placental development.

Testing For Territoriality

For many animals, territory defense is difficult or impossible to document but patterns of home range overlap can be documented. Such patterns of overlap can often be used to deduce territorial behavior. Table 3.1 gives overlap index results for territorial wolves in northeastern Minnesota (Mech, personal communication, unpublished data), for adult female black bears in North Carolina and in Minnesota that appear to differ in territorial behavior (Powell et al. 1997, Rogers 1977), and for intrasexually territorial stone martens (Martes foina) in Italy (Genovesi and Boitani 1997 Genovesi et al. 1997 Genovesi personal communication, unpublished data).

QoL Research or Clinical Instrument

Even if larger trials are needed to confirm these results, these conclusions are very important. Collecting information by routine use of QoL scores can help physicians to follow patients over time and plan treatment on specific areas affected by the disease. In our experience (unpublished data), weight loss and gastrointestinal symptoms (such as nausea and vomiting) are early predictors of tumour recurrence and, sometimes, they start 3-4 months before radiological finding or increase in neoplastic markers.

Mixed Secondary Structures

The helix and hairpin structures considered in the preceding sections are conformationally well defined and can be used in a modular fashion to create supersecondary structure motifs. Assembly of modules can be achieved by the use of linking segments that control orientation, or by building favorable interactions between the modules. Thus far, the success in building mixed a P structures has been limited. Figure 11 shows an example of a 17-residue helix-hairpin peptide that has been characterized in crystals (46). In this case, an octapeptide hairpin has been linked to a heptapeptide helix using a 2-resi-due Gly-Gly linking segment. The conformations of both the modules are maintained in the target peptide. Figure 12 provides a counterexample in which profound changes in the structure of the module may accompany its incorporation in the longer sequence. In this case, a helical module (Boc-Leu-Aib-Val-Ala-Leu-Aib-Val-OMe) and a P-hairpin nucleated by an Aib-DAla turn were connected by...

Polyamine Metabolism is Disturbed in Prostate Cancer

The recent development of the microarray technology has allowed analysis of expression levels of thousands of genes in one experiment. In a study to identify genes and pathways that are differentially expressed between androgen-dependent and androgen-independent cancer cells, Jenster et al. from the Department of Urology of the Erasmus MC in Rotterdam performed microarray experiments, using GeneChips that harbored approx 4000 known genes and 3000 expressed sequence tags. On each chip, two batches of RNA were compared by competitive hybridization with the glass-spotted complimentary DNAs. Comparing the expression of ODC, AdoMetDC, and SSATgenes in androgen-responsive LNCaP cells cultured in the absence or presence of 1 nM di-hydrotestosterone (DHT) for 16 h, it was observed that androgen stimulated the expression of these enzymes twofold to fourfold (Jenster et al., unpublished).

Insulin and Amino Acids

It is well established that insulin-like growth factor-1 (IGF-1) plasma concentrations reflect the intake of dietary protein 17, 18 . In a study comparing growth of infants fed formulas with different protein concentrations 19 , determinations of plasma concentrations of IGF-1 were obtained 20 . One formula contained protein at a concentration of 2.39g 100kcal and the other at 1.90g 100kcal. Similar determinations were made in a group of exclusively breast-fed infants participating in another study (unpublished). The results summarized in table 2 show that plasma IGF-1 concentrations were similar in the 2 groups of formula-fed infants and in the breast-fed infants at 1 month of age. However, by 4 months of age, the IGF-1 concentrations of breast-fed infants and infants fed the lower protein formula had declined significantly, whereas they remained unchanged in infants fed the higher protein formula. At 4 months of age, the IGF-1 levels of breast-fed infants were significantly lower...

Clonal Variations Of Phenotype

20 kbp deletion close to the rrnC locus. Moreover, PAO1 stocks maintained at different laboratories are not identical in phenotypic traits, a spectacular example being the differential virulence in infection models even though the strain had been originally obtained from the same public collection (own unpublished data). During storage and subculturing the PAO1-derived isolates apparently diversified by inversion, deletion, and point mutation.

Background and Epidemiology of Hepatitis Viruses in Correctional Settings

Persons with chronic HBV infection serve as the primary source of infection for others (McQuillan et al., 1999 CDC, 2005). The majority of persons with chronic HBV infection are asymptomatic, and one third have no evidence of liver disease, despite high levels of viral replication (Lee, 1997). Chronic HBV infection can lead to cirrhosis and HCC. Lifetime risk of death from chronic liver disease or HCC is 15-25 (Beasley, Hwang, Lin, & Chien, 1981 Beasley, 1988 Chang et al., 1997 McMahon, 1997 McMahon, Holck, Bulkow, & Snowball, 2001). Rates of progression to cirrhosis and HCC are approximately 25 for persons who acquire infection during childhood and 15 for persons who acquire infection at older ages. Other factors that influence rates of progression include HBeAg status coinfection with HDV, HIV, HCV and alcohol abuse (Rizzetto, 1983 McMahon, 1997 Ockenga et al., 1997 Zarski et al., 1998 Monto & Wright, 2001 Gao, 2002). HBV-related liver disease and HCC cause approximately 4000-5000...

PPeptide Synthesis Using Solid Phase Methods by Fmoc Based Chemistry

Synthesis of P-peptides longer than 6 residues have not been straightforward because of the difficulty of removing the Fmoc-protecting group at the fifth through seventh residues (44,54). However, the difficulty in Fmoc removal at these residues appears to be sequence dependent (unpublished results). Several modifications to the standard protocol have been developed to overcome this apparent difficulty including alternative deblock procedures involving both piperidine and DBU (54) and elevated temperatures (44). These modifications are not reflected in the procedures provided in this section, but may be implemented if necessary as described in the notes (see Note 4).

Methods Of Analysis

The past several years have seen dramatic advances in technology and equally dramatic reductions in cost. Both genomic and proteomic methods are now available at costs that allow an average small laboratory to begin performing such experiments. The amount of material required for such an analysis has also become much less than was previously necessary, thereby making these experimental approaches feasible even when the number of stem cells available is limited. For example, single-nucleotide polymorphism analysis to examine overall allelic variability using a 500-K chip set costs about 500. Likewise, a genome-wide gene expression profile using an Illumina 48,000 transcript chip costs about the same amount. Mitochondrial analysis and nuclear run-on assay expenses are also in the same ballpark. Even more importantly, the requirement of material has been dramatically reduced. We estimate that one can perform the entire battery of tests (excluding proteomic analysis) with about 2 million...

Jasmonates in Plant Development and Senescence

The capacity of jasmonates to down regulate photosynthetic genes (Creelman and Mullet, 1997) may also be one determinant in the onset of senescence. In this respect it is interesting to note that barley plants exhibit high JA levels in tissues undergoing senescence (unpublished data from our laboratory), and that ELIPs, considered to be protective against high-light conditions, are down regulated by JA (Wierstra and Kloppstech, 2000).

Bruce Dicka Jane Fergusona Venkatraman Chandra Moulia Loretta Brabinb Subidita Chatterjeec David A Rossd

Methods We identified published and unpublished studies and reports from developing countries that provided information about interventions designed to increase young people's use of health services. The studies were classified into six different types based on whether they included some or all of the following characteristics training for service providers and clinic staff making efforts to improve the quality of the facilities implementing community activities to generate demand and support for the services and involving other sectors, notably schools and the media. The levels of evidence required to make decisions about policies and programmes were defined for each of these types.

Data Mining Chromosome Mapping Pathway Analysis And Data Representation

Empirically, we have determined that comparison across platforms is fraught with peril. Only positive results can be considered, and negative results are generally not interpretable. For example, ESC-derived NSC samples were taken and examined by MPSS and Illumina bead arrays, and a concordance of around 50 was shown, whereas the concordance rate for the sample run on a second Illumina array was close to 95 (unpublished observations). Even with such a comparison, only the presence or absence of an expression pattern can be considered, and no attempts to compare expression levels between different methods should be made. We have also determined that amplification tends to provide a different pattern than using unamplified RNA, even if the same sample is used. This appears to not be the result of operator error, because the amplification of different biologic replicates performed at different times yields more similar results than comparison between amplified and unamplified samples....

Cooperativity Between Active Sites

Flexibility of the proteasome underlying such cooperative effects is also apparent from the effects of activating agents like SDS and poly-lysine or natural activators like PA28 on peptidase activities. The enhanced frequency of coordinated dual cleavages in longer peptides observed upon binding of the PA28 activator to mammalian proteasomes48 largely rules out that facilitated access of substrates to the active sites in the interior is the only reason for activation, as one could argue in the case of SDS or heat treatment. The possibility of such long distance modulation of proteolytic activities is also evident from the fact that certain mutations in a given a type subunit can strongly activate distinct peptidase activities and at the same time inhibit others (W. Heinemeyer, unpublished results).

Transgenic Plant Lines

There is little information on transcription and translation activity in maize gametes and zygotes. The competence of maize gametes and zygotes to express stable integrated transgenes was shown in our laboratory with plants transgenic for the gfp gene. The same gfp vector (35S gfp, 67 ), as used for microinjection experiments, was introduced. It was optimised for high expression levels of GFP in monocotyledonous plants and codes for a plant codon usage optimised S65T version of the gfp gene. In female gametophytes the 35S gfp construct was expressed. Egg cells, synergids, and central cells showed GFP fluorescence, whereas no fluorescence was detected in transgenic male gametes 72 . After fertilisation of non-transgenic egg cells and central cells with transgenic sperm cells expression of the transgene was induced early after fertilisation, and GFP was detected in zygotes and early endosperm (S. Scholten, unpublished results). The 35S promoter construct was active in egg cells, central...

Bone marrow transplantation and immunosuppressive treatment

Figure 11.4 Successful thrombolytic therapy in PNH. (a) Magnetic resonance venography in a 29-year-old woman who presented with acute Budd-Chiari syndrome, and who also had a thrombus partially occluding the inferior vena cava. (b) Repeat study after a 48-h infusion of tissue plasminogen activator (from R Thertulien et al., unpublished).

Aging Pm In The Laboratory

Terminology aside, the data are reasonably consistent in showing smaller age-related deficits with focal than with nonfocal cues (e.g., Maylor et al., 2002 McDaniel et al., chap. 7, this volume Rendell, McDaniel, Forbes, & Einstein, 2007 Salthouse et al., 2004). However, the data are less consistent on the question of whether age deficits can be eliminated altogether with focal cues. For example, Salthouse et al. (2004) observed age deficits in their PM tasks with focal cues (namely, drawing classification and concept identification ) and several of my own (unpublished) attempts to replicate Einstein and McDaniel's (1990) classic findings of no age deficits with focal cues have failed. Some reasons for these discrepancies are discussed later.

Sample Resolution And Information Obtained

Mately 1 (identical diet Krebs 1989 Litvaitis 1992). During an investigation of carnivore interactions (Litvaitis 1992), the overlap between coyotes (Canis latrans) and bobcats (Felis rufus) was 0.83. Because adult male bobcats are 50-100 percent larger than adult females, male bobcats are able to exploit larger prey and thus may be more similar to coyotes, which are substantially larger than female bobcats. To examine this possibility, samples from bobcats were separated into two classes based on body mass and diet overlap between each size class of bobcats and coyotes was calculated. As suspected, overlap was substantially greater between adult male bobcats and coyotes (0.95) than between coyotes and female and juvenile bobcats (0.78) (Litvaitis unpublished data). This example suggests the benefits of obtaining greater sample resolution and not averaging samples from a population where resource segregation has occurred.

Abnormally Developing Embryos

Arrested embryos and embryos of women of advanced maternal age ( 35 years) were found to have a higher incidence of numerical aberrations for chromosomes 18, X and Y when compared to controls (Munne et al., 1993). Multinucleated blastomeres, most commonly binucleate cells, have also been observed in cleavage stage embryos, especially in arrested embryos (Munne et al., 1993) and apparently more readily in certain patients (Harper, unpublished observation). The formation of these cells may be the normal pattern of trophectoderm development as a precursor of syncytiotrophoblast formation, for it also occurs in cattle (Hare et al., 1980), pig (Long & Williams, 1982) and merino ewes (Murray et al., 1986). Although it has been suggested that these binucleated blastomeres were precursors for mural trophectoderm giant cells, they in fact appear at cleavage stages before trophectoderm differentiation. While it is possible that binucleate cells form from asymmetrical cytokinesis so that one...

Automated blood pressure measuring devices

The need for a standardized protocol became apparent in the late 1980s after the widespread introduction of automated devices to the market. Various protocols have since been published with the aim of establishing a minimum standard of accuracy. The Association for the Advancement of Medical Instrumentation (1993), the British Hypertension Society (O'Brien et al., 1993a) and the European Society for Hypertension (O'Brien et al., 2002) all published protocols and both Germany and Australia have unpublished recommended national standards.

FRET Measurements to Determine the Proximity of Proteins to Each Other

The second method, to measure the decrease in donor's fluorescence in the presence of acceptor, usually requires taking measurements before and after photobleaching of the acceptor, because physically removing the acceptor is often impossible for in vivo experiments. Bleaching the acceptor in a manner that is not toxic to the cell is difficult and inefficient, particularly with CFP and YFP as the donor-acceptor pair (unpublished results). Still the approach

Variations On The Methyleneinterrupted Double Bond

Finally, the plant sphingolipid desaturases (65) represent a possible ancestral fusion of an N-terminal cytochrome b5 with a A6-desaturase first described in borage (Boragio officinalis) seeds. Borage seeds, in common with a few other plants such as evening primrose (Oenothera biennis) and black currant (Ribes nigrum), contain oils rich in y-linolenic acid (18 3 A6,9,12). Given the primary sequence homology between the ubiquitous plant sphingolipid desaturases and the borage A6 desaturase, it seems probable that the borage-type A6 desaturases represent enzymes that have diverged in substrate specificity from the A8 A6 sphingolipid desaturase family. Curiously, the borage A6 desaturase is extremely effective at producing up to 50 GLA in transgenic soy seed (S. Coughlan, unpublished results) but not in transgenic Arabidopsis or canola seeds (66).

Translational Control by Paips PABP Interacting Proteins

Two proteins that strongly interact with PABP and affect translation were identified by Far-Western interaction screening 316, 317 . Paip1 is a 56 kDa protein, which activates translation in vivo. It is homologous to the central region of eIF4G and interacts with eIF4A 316 (Fig. 7.2-11A). Paip1 is also involved in mRNA turnover, as it is found in a protein complex with PABP that recognizes the major protein-coding region determinant of instability (mCRD) of the c-fos proto-oncogene mRNA 318 . Paip2, a much smaller protein (MW 14 kDa), preferentially inhibits the translation of mRNAs containing a poly(A) tail, and also IRES-containing mRNAs that are eIF4G-dependent 317 . Paip2 inhibits the formation of 80S initiation complexes 317 . Although it is possible that Paip2 inhibits the conversion of 48S preini-tiation complexes to 80S initiation complexes 289 , Paip 2 also partially inhibits the formation of 48S preinitiaition complexes (Kahvejian and Sonenberg, unpublished). In an...

Review of Literature on Prescribing Restrictions

Table 6.1 summarizes four studies of the financial effects of hospital formularies. One unpublished study (Sullivan and Hazlet33) suggests that restrictive hospital formularies are associated with reduced drug cost without increasing length of stay (a surrogate for total cost per admission). The others show that reductions in drug cost associated with hospital formulary restrictiveness result in part from cost shifting, i.e., reducing expenditures in one account (the drugs budget) at the expense of another, e.g., the nursing budget.

The Procedures of Gait Training

The primary emphasis in gait training is on the patient's trunk. Approximation through the pelvis during stance and stretch reflex to the pelvis during swing facilitate the muscles of the lower extremities and the trunk (S. S. Adler, unpublished, 1976). Proper placement of the hands allows the therapist to control the position of the patient's pelvis, moving it toward an anterior or posterior tilt as needed. When pelvic motion and stability are facilitated the legs can function more efficiently. Our hands can also be on the shoulder and on the head for stabilizing or facilitating trunk rotation.

Sequence polymorphism and falsenegative results

Recently, we compared a 5' nuclease assay targeting the parainfluenza 3 nucleocapsid protein gene (WhSl-para3-5N unpublished data) with a previously described 5' nuclease assay targeting the parainfluenza 3 hemag-glutinin-neuraminidase gene (HN-para3-5N Watzinger et al., 2004) for the detection of parainfluenza 3 in nasopharyngeal aspirate specimens collected from a local patient population. During the course of the study, 34 positive samples were identified with both assays detecting 33 of these. The cycle threshold (Ct) values for these 33 positive samples were comparable in both assays, with an average difference of only 1.1 cycles. However, one sample was negative by the WhSl-para3-5N assay but was positive by the HN-para3-5N assay, providing a Ct value of 25. Such a low Ct value indicates that the viral load was relatively high in this specimen and therefore the false-negative result could not be explained by the differences in the detection limit of each assay. The presence of...

Implications of the Two Step Model for Polyamine Transport and Homeostasis

Evidently, the suppression of net polyamine uptake exerted by AZ could theoretically involve an interaction of AZ with either plasma membrane or vesicular transporters, or both classes of carriers. Interestingly, preliminary observations using Spd-C2-BODIPY suggest that cytosolic levels of the fluoroprobe dramatically increase under conditions whereby AZ induction is prevented (Gagnon, B., Soulet, D., Rivest, S., Audette, M. and Poulin, R., unpublished results). These data suggest that inhibition by AZ may result mainly or exclusively from their interaction with the plasma membrane polyamine carrier.

Pathologic Evaluation of the Sentinel Lymph Node in Malignant Melanoma

With the advent of selective lymphadenectomy, pathologists have recognized that a detailed method for assessing these nodes is needed, but there is little concurrence in the general literature as to exactly what constitutes an ideal protocol. The standard method of evaluating lymph nodes for metastatic tumor involves submitting 1-2 representative sections from the center of the lymph node for H&E staining. This method is sensitive enough to detect 1 abnormal cell in a background of 104 normal cells.5 This sensitivity can be increased 10-fold by using immunohistochemical staining.6 Immunohistochemical stains which have been particularly useful in melanoma diagnosis include S-100 and HMB-45. S-100 is an acidic, calcium-binding protein so named because of its solubility in 100 ammonium sulfate. It was first identified in mammalian brain tissue, and subsequently identified in tumors of neural origin. Extensive studies have now shown this protein to be present in a wide variety of...

Large Conformational Changes Associated with RF2 Binding to the Ribosome

This finding fits nicely with the wealth of biochemical data implicating the GAC as an interaction site for RF1 and RF2. Preliminary footprinting studies performed in the early 1990s using RF2 identified a change in the protection pattern within the 2660 region (H93) of the 23S rRNA (C.M. Brown and W.P. Tate, unpublished results). Cleavages were also identified within this region from tethers located at position 229 in RF1 76 and 650 of EF-G 74 . In addition, position 229 of RF1 cleaves within the L11-binding region, both in H43 and H44 76 , as do tethers from positions 650 and 655 in domain V of EF-G, which cleaved within the 1095 and 1067 stem-loops, respectively 74 . This region is the target for a family of thiazole antibiotics such as thiostrepton and micrococcin, both of which have been shown to inhibit elongation (see Chap. and termination factor activity 81 . When 70S ribosomes were treated with anti-L11 (and anti-L16), RF-dependent peptidyl-tRNA

Whole Library Phenotypic Screens

We found that it is feasible to carry out phenotypic screens of the entire mutant library. On the basis of comparisons with earlier studies of two well-defined processes (twitching or swarming motility and growth on minimal medium), the approach appears to provide virtually complete lists of genes in which loss-of-function mutations confer the phenotypes of interest.6 The collection has more recently also been screened for mutants showing reduced growth under anaerobic conditions and altered sensitivity to the antibiotic tobramycin (I. Thaipittsukul S. Lee, unpublished results). In each case, the protocol followed was to spot small aliquots of each mutant onto appropriate medium using a 384-pin pronging device, followed by incubation, digital imaging, and one or more rounds of scoring by visual inspection.

Inclusion and exclusion criteria

A number of review articles on HIV prevention interventions that focused on young people (1-6) and developing countries (7) were scanned for relevant studies. All reference sections of peer-reviewed papers identified during this literature search were checked for additional articles of interest. Finally, 12 experts working in the field were contacted to identify unpublished research.

Disability Specific Costs Beyond Early Childhood

Although few studies have tabulated the medical care costs of preterm birth in early childhood, fewer still have analyzed the medical costs beyond early childhood for those born prematurely. Lifetime estimates of cost, however, have been made for individuals with certain conditions and developmental disabilities associated with preterm birth and LBW, such as specific birth defects (Waitzman et al., 1996), CP (CDC, 2004c Honeycutt et al., 2003 Waitzman et al., 1996) and MR, HL, and VI (CDC, 2004c Honeycutt et al. 2003). Prevalence estimates of developmental disabilities in a Centers for Disease Control and Prevention (CDC) study (CDC, 2004c) were drawn from MADDSP, which CDC established in 1991 to identify children with these developmental disabilities (Yeargin-Allsopp et al., 1992). The CDC study, together with unpublished tabulations of differences in the prevalence of each developmental disability by ges-tational age from MADDSP (see Table 12-8), permitted the assessment of the...

Enzymes of Phosphatidylethanolamine Biosynthesis

When a cDNA encoding the human CTP phosphoethanolamine cytidylyltransferase was expressed in rat hepatoma cells, the in vitro enzymatic activity was increased by three-fold. However, the incorporation of 3H ethanolamine into PE was only modestly increased and the PE content of the cells was unaltered (S. Stone and J. E. Vance, unpublished data). Interestingly, this overexpression of CTP phosphoethanolamine cytidylyl-transferase activity neither inhibited PS synthesis nor reciprocally inhibited the production of PE from the PS decarboxylation pathway (S. Stone and J. E. Vance, unpublished data).

Tests of the Theories

At the foundation of the evolutionary theories of senescence are the theoretical predictions of Medawar (1952). He suggested that in a natural environment a population would experience a high level of random, age-independent, mortality due to harsh environmental conditions. As a result, there is an ever-decreasing proportion of individuals that survive to reproduce at older ages. Harmful genes whose time of expression is beyond the natural range of life span for a particular species, can accumulate in a population with little or no check because the effect of mortality from external causes will be to reduce the force of selection in each successive portion of the life span. As a result, natural selection is ineffective at preventing an accumulation of deleterious genes that affect late-ages because they have a minimal impact on the population. Medawar concluded that aging is the result of the cumulative expression of deleterious genes in individuals that live longer than the average...

Predictability Of Behavior

Measures of uncertainty can be a valuable tool in assessing the predictability of responses of individuals to others. Again, we explore an example from our work on wood mice (Stopka and Macdonald, unpublished data). For example, all combinations of five male and four female wood mice kept under video surveillance in an enclosure with abundant nest boxes sometimes denned communally, although there was no thermoregulatory need to huddle. It was therefore surprising that cohabitants were often agonistic to each other. One of the most common categories of interaction was approach and nose-to-nose contact, categorized by Bovet (1972a, 1972b) as amicable, but lag sequential analysis revealed that although this (nose-to-nose) behavior pattern was indeed sometimes a precursor to flank-to-flank or nasoanal contact, on other occasions it was followed by attacking or chasing. Therefore, nose-to-nose encounters are defined by their context within a sequence. Sequential analysis revealed that...

Arrangement of ATPases Within the Regulatory Complex

These proteases, hexameric rings of ATPase subunits attach to both ends of stacked hexa-(ClpQ) or heptameric (ClpP) proteolytic rings. Another well-characterized member of the AAA family, p97 or valosin-containing protein (VCP) also forms hexameric rings.84 Hence, it is reasonable to assume that a hexameric ring of S4-like ATPases sits atop the peripheral rings of the proteasome. Nonetheless, this model is difficult to reconcile with published images of the 26 S proteasome.21,85,86 A hexameric ring of roughly 45 kDa proteins would presumably have a diameter larger than the underlying proteasome ( 11 nm) and extend beyond the edges of the cylinder. By contrast, a tetramer of ATPases would probably have a diameter less than the proteasome, more consistent with the images of Peters et al and Yoshimura et al.21,85 We recently observed that tetramers of ATPase subunits (S4, S6, S7, and S8 or S4, S8, S6', and S10b) can be coimmuno-precipitated with the non-ATPase subunit 5b after...

Regulation of Transgene Derived SSAT Gene Expression

Activity was induced by treating transgenic mice overexpressing SSAT transgene under the control of mouse metallothionein I promoter with A A -diethylnorspermine (DENSPM) (11). The latter induction most likely occurred at a level of gene expression downstream of mRNA accumulation, as the transgene was driven by a heterolo-gous promoter not supposed to be induced by polyamine analogs. Some recent evidence indicates that alternative splicing of pre-mRNA may be involved in the regulation of SSATgene expression (12,13). Our unpublished findings have likewise indicated that polyamine analogs influence the splicing of SSAT pre-mRNA. As expected, transgenic mice overexpressing SSAT and fetal fibroblasts derived from them are much more sensitive to the toxic action exerted by polyamine analogs (10,14).

DNA Arrays in The Research of Atherosclerosis

Macrophage-rich shoulder areas of human atherosclerotic lesion were laser mi-crodissected and gene expression profiles were compared to normal intima. Upre-gulation of several macrophage-specific genes e.g. IL-4 and additionally, upregula-tion of many known e.g. pancreatic lipase and unknown genes were detected (Tuomisto et al 2002, unpublished results).

Possess Abnormally Short Telomeres

In agreement with the previous telomere length analyses, this method revealed shorter telomeres in primary prostate cancer cells, when compared with normal epithelial cells, in 15 (88 ) of 17 cases (64). This analysis is currently being extended to include more primary tumors as well as local and distant metastatic lesions. To date, 86 of 50 primary tumors examined contained exclusively or predominately shorter-than-normal telomeres, whereas only 6 had telomeres in the normal range. In no case was abnormal telomere lengthening observed in these primary tumors. Similarly, 84 of local metastases (pelvic lymph nodes) displayed telomere shortening, whereas a small subset (5 ) displayed normal or longer-than-normal telomeres, and 11 exhibited notable telomere length heterogeneity. A smaller proportion of distant metastases, 57 (56 of 98 metastases from 28 cases), had short telomeres. However, in 11 cases in which distant metastases could be matched to their...

Structure Based Optimization of Fragment Hits

When the binding of one or more molecular fragments has been determined in the proteinactive site, this provides a starting point for medicinal chemistry to optimize the interactions using a structure-based approach. The fragments can be combined onto a template or used as the starting point for growing out an inhibitor into other pockets of the protein (Fig. 10.5). The potency of the original weakly binding fragment can be rapidly improved using iterative structure-based chemical synthesis. For example, in one of our lead discovery programs targeted against the cancer target cyclin-dependent kinase 2, we identified an initial fragment, AT381 (MW 134), which exhibited an IC50 of 1 mM in an enzyme assay. Using the crystal structure of AT381 bound to CDK2 we were able to improve the potency more than 100000-fold by synthesizing only 30 analogues (Fig. 10.6). The resulting compound, AT3851, had an IC50 30 nM and was generated using an iterative cycle of design and synthesis (unpublished...

Matrix metalloproteinaseactivated prodrugs

Was demonstrated, presumably due to lack of drug release from the liberated tripeptide-drug conjugate. Although the specific mechanism of drug release was not addressed, polymeric prodrugs of Mmc containing the peptide sequences Gly-Phe-Ala-Leu and Gly-Phe-Leu-Gly-Phe-Leu between polymer and drug have been reported to release drug in the presence of collagenase IV (MMP 2) (Soyez et al., 1996a). MMP activation of a low-toxic hexapeptide (Pro-Gln-Gly-Ile-Mel-Gly) that contained the parent drug melphalan (Mel) has also been described (Timar et al., 1998). MMP cleavage generated Ile-Mel-Gly, which was more toxic than the hexapeptide, but liberation of free melphalan was not reported. An octapeptide conjugate released Ile-Ala-Gly-Gln-Dox upon MMP activation (Kratz et al., 2001). Again, liberation of unconjugated parent drug was not observed. Not only a range of MMP-activated prodrugs that has been published but also some unpublished MMP-cleavable prodrugs prepared in our own lab release...

Sources for Further Study

Karen Horney A Psychoanalyst's Search for Self-Understanding. New Haven, Conn. Yale University Press, 1996. A biography of Horney that places her theories squarely within the context of her life history. Written by the editor of the papers unpublished during Horney's lifetime (The Unknown Karen Horney, 2000). Quinn, Susan. A Mind of Her Own The Life of Karen Horney. Reading, Mass. Addison-Wesley, 1988. This biography is an excellent source of information about Horney's personal and professional life. Much of it is devoted to her female psychology. Easy to read contains photographs, biographical essays, extensive source notes, and a complete list of Horney's work. Rubins, Jack L. Karen Horney Gentle Rebel of Psychoanalysis. New York Dial Press, 1978. The first biography of Karen Horney. Thorough and well documented includes detailed discussions of Horney's theories on women. Lengthy but well organized. Can be read by the college or high school student.

Isolation and Characterization of HR Markers

Molecular cell death markers can constitute useful tools to compare different kinds of cell death and to address the question of the existence of common steps or crosstalks between them. For example, HSR203J has been shown to be activated during HR cell death induced by different pathogens, upon HR-like cell death induced by the ectopic expression of the bO proton pump and during some disease-related cell deaths (Pontier et al., 1994, 1998b, 1999). In contrast, no activation of its promoter has been detected during leaf senescence or differentiation of tracheary elements (Pontier et al., 1999 D. Pontier, unpublished results). Similarly, the Arabidopsis SAG12 gene expression is highly associated with leaf senescence and is not detected after salicylic treatment or in the dying cells during the HR in tobacco (Weaver et al., 1998 Pontier et al., 1999 Quirino et al., 1999). These two genes may thus be part of signaling steps specific for HR cell death and senescence respectively. However,...

Transgenic Gtpch Deficiency and Overexpression in Mouse

Hypertension Mouse

We first demonstrate that congenital BH4 deficiency in the hph-1 mouse results in the development of pulmonary hypertension and vascular remodelling in the adult under normoxic conditions and exacerbates the response to hypoxia 43 . Conversely, augmentation of endothelial BH4 biosynthesis in the GCH transgenic mouse (in which GCH-overexpression is target to endothelial cells) protects against the development of hypoxia-induced pulmonary hypertension and vascular remodelling during 4 weeks of hypoxia (unpublished data). Selective restoration of endothelial BH4 levels in the hph1 strain by intercross with the GCH transgenic mouse rescues the effects of systemic BH4 deficiency. A striking quantitative correlation was observed between lung BH4 levels and the development of pulmonary hypertension across the 5 genetic mouse models evaluated in our studies, suggesting that BH4 bioavailability controls both pulmonary vascular tone and structural remodelling in a dose-dependent manner 43 ....

Evidence for Unequivocal Genetic Effects

We have used this general drug-by-strain approach in unanesthetized animals. As studies in NOS knock-out mice have shown that the nitric oxide system can influence post-hypoxic ventilatory behavior 48,51,49 , the question arises whether nitric oxide synthase (NOS) blockade would similarly effect ventilatory behavior in two rat strains. After vehicle, post-hypoxic decline in frequency and minute ventilation is not evident in the Sprague Dawley strain 85 . NOS blockade resulted in a greater increase in resting ventilation in the Sprague Dawley than in the Brown Norway strain, but produced post-hypoxic frequency and ventilatory decline in the Sprague Dawley strain 84 . We have observed a similar effect using a selective neuronal NOS (NOS-1) blocking agent (7-nitroindazole or 7-NI, 60 mg kg) (K. Strohl, unpublished findings). The results indicate that the various strains do not utilize the NO pathway in the same way with regard to the regulation of ventilatory behavior.

Iron Uptake Via Siderophores 21 Siderophores

Under iron deficient conditions, all fluorescent Pseudomonas are characterized by the production of yellow-green, water-soluble siderophores called Pvds. Pvds form a wide class of mixed siderophores with a great variety of structures and a high affinity for iron, with association constants as high as 1032 M-1.5 These siderophores are effective at acquiring iron from transferrin102,166,167 and lactoferrin167 and this has obvious implications vis-a-vis in vivo growth and pathogenesis. Mutants with deficiencies in Pvd biosynthesis or transport are effectively avirulent in animal models of infection.102,151 In addition to their high-affinity iron chelators, fluorescent pseudomonads are known to produce lower-affinity siderophores, such as pyochelin, a derivative of salicylic acid, salicylic acid itself,99,161 and quinolobactin.111 These compounds can be considered as secondary siderophores since they are usually produced in much lower amounts than Pvds (a few mg l vs. 100-200 mg l for the...

Fluorescence Activated Cell Sorting FACS

Excitation (dashed lines) and emission (solid lines) spectra of wt GFP (gray lines) and GFP-S65T (black lines) Although precise data are not available, the excitation and emission spectra of RSGFP appear to be very similar to those of GFP-S65T (21, Clontech, unpublished observations). The emission data for wt GFP were obtained with excitation at 475 nm. (The emission peak for wt GFP is several-fold higher when illuminated at 395 nm the major excitation peak of wt GFP , However, the filter sets commonly used for GFP fluorescence microscopy and the argon ion lasers used in most FACS machines excite around 450-500 nm). The emission data for GFP-S65T were obtained with excitation at 489 nm, which corresponds closely to the excitation peak of GFP-S65T. (The data in this figure are derived from ref. 23.)

Androgen Independent PSA Secretion

Secretion Androgene

ErbB-2 can up-regulate PSA secretion at two levels the transcriptional level and the post-translational level in the secretory pathway. In C-81 cells in which ErbB-2 is activated by hyper-tyrophosphorylation, the level of PSA mRNA is elevated in the steroid-reduced condition, compared to C-33 cells,27'43 which may be due in part to the activation of the PSA promoter by ErbB-2 via ERK MAPK and AR.69 Alternatively, ErbB-2 may activate MEK1, leading to the up-regulation of a transcription factor(s) that subsequently activates the PSA promoter.76 ErbB-2 may also activate the secretory pathway of PSA in those cells (M. S. Lee and M. F. Lin, unpublished observations). Nevertheless, further experiments are needed.

Hurthle Cell Carcinoma

Our unpublished series of 62 patients had a median age at diagnosis of 59 years and median follow-up of 5 years a neck mass was the commonest mode of presentation (85 ). Tumors were larger than 4 cm in 31 , displayed extrathyroidal extension in 20 , and distant metastasis at presentation in 15 . Overall survival at 10 and 20 years was 64 and 37 , respectively. Multivariate analysis identified nodal disease, distant metastasis, and tumor size to be associated with an increased cause-specific mortality (P 0.01). Extent of surgery improved cause-specific survival, whereas radioactive ablation had no effect on survival but EBRT achieved symptomatic benefit in 11 of 14 distant metastases. None of 17 metastatic sites concentrated 131I.

Mechanisms of Ozoneinduced Accelerated Leaf Senescence

Ethylene And Senescence

There are many mutants of Arabidopsis that lack ethylene perception, due to mutations in receptors or in down stream signaling components, e.g. etr1-3 (Guzman and Ecker, 1990). No difference in SAG expression was found between the Columbia ecotype of Arabidopsis and etr1-3, following O3 treatment (Miller, Arteca and Pell unpublished data). This work suggests that ethylene perception by the etr1-3 receptor is not required for induction of the O3-responsive SAGs. The role of ethylene perception cannot be completely ruled out, due to the redundancy of ethylene perception in Arabidopsis. Other ethylene insensitive mutants are being studied to better define the role of this hormone in O3-induced SAG expression (Miller, Arteca and Pell, unpublished data).

Prehistoric and Preindustrial Eras

Most of this increase was due to the reduction of infant and child mortality. It used to be the case, for example, that remaining life expectancy at age 1 was greater than at birth, because the toll of infant mortality was so high. The difference between premodern periods and today is less stark if we consider life expectancy at higher ages. Instead of the tripling of life expectancy at birth, remaining life expectancy at higher ages has roughly doubled over the course of human history. At age 10, for example, life expectancy (i.e., expected years after age 10) may have moved from the around 30 to 33 years to almost 70 years (Thatcher AR. Life tables from the Stone Age to William Farr, unpublished manuscript, 1980). At age 50, it may have gone from around 14 years to more than 30 years (17).

Furin Regulation By Hypoxia 81 Tumor Hypoxia

Hif And Tgf

Several studies suggest that furin-induced expression within tumors supports tumoral development through the increased bioavailability of pro-tumorigenic mediators. Interestingly, the hypoxia HIF-1-dependent increase in furin gene expression was found to be associated with increased maturation activation of the angiogenic tumorigenic mediators MT1-MMP and TGF 1 19 . Both mediators are well characterized furin substrates that have been found to profoundly affect many aspects of tumor progression 14, 17, 64 . MT1-MMP, through proteolytic events, regulates various cell functions, including extracellular matrix turnover, promotion of cell migration and invasion. MT1-MMP acts either through direct degradation of extracellular matrix components or indirectly by activating MMP-2 65 . In addition, these metalloproteinases are involved in the angiogenic process by mediating endothelial cell invasion as well as the release or the activation of growth factors 66, 67 . TGF 1, in turn, creates a...

Finding Complex Modification Pathway Genes

Queuosine Biosynthetic Pathway

I am grateful to Henri Grosjean for his constant encouragement and help in getting started in the tRNA modification field and for his careful revision of this manuscript. I thank Dirk Iwata-Reuyl for significant contributions to the manuscript and for providing several figures. I thank Glenn Bjork, Louis Droogmans and Tsu-tomu Suzuki for unpublished information and Charles Lauhon for suggestions. This work would not have been possible without the support of Paul Schimmel.

Mayapple as a Study Organism

Rhizome Diagrams

Mayapples are perennial clonal herbs of deciduous forests of the northeastern United States they are found growing in discrete patches or colonies containing one or more genotypes (genets) (Policansky, 1983 Whisler and Snow, 1992). Mayapples are morphologically simple and have a limited number of developmental options that can directly influence their demography (their pattern of birth, death and ramet population increase) and their life history expression (whether they are sexually reproductive or not). These developmental options include annual decisions regarding rhizome branching, a developmental process that results in an increase in the number of ramets present, and whether to differentiate a vegetative or a sexual aerial shoot (Fig. 17-1) (Watson et al., 1997). Nitrogen and carbon availability influence the pattern of mayapple development and, hence, demographic and life history expression (Benner and Watson, 1989 de Kroon et al., 1991 Landa et al., 1992 Geber et al., 1997...

Multilevel Interactions The Ecosystem

The proteases released in the soil by the resident fauna and microorganisms, or those found in our own digestive system could lead to unwanted inhibitory effects susceptible to affect natural equilibria in the ecosystem, or to alter the nutritive quality of plant food products (see Fig. 9.2 also see Chapters 8, 14 and 18). Transgenic potato plants expressing OC-I, for instance induce compensatory processes in the natural predator Perillus bioculatus via its prey, the Colorado potato beetle (E. Bouchard et al, unpublished observations), in agreement with the occurrence of digestive cysteine proteinases sensitive to this poorly-efficient cystatin in the predator.27 These compensatory responses did not cause developmental delays in the predator, but it cannot be excluded that using PIs more efficient than OC-I could eventually have negative effects,28 and thus affect the effectiveness of this useful biological control mean in the field.

The Anterior Visceral Endoderm Domain Is Restricted By The Extraembryonic Ectoderm

Tracing the clonal origin of AVE cells is a delicate matter, because implantation cannot be fully reproduced in vitro, and conceptuses can only be cultured over a short period. This has been analyzed by a cell-labeling approach. Single precursors of the visceral endoderm were targeted before implantation, at the blastocyst stage, by the microinjection of mRNA encoding a fluorescent protein (Figure 10.4, A). Positive blastocysts were transferred into the uterus of a foster mother to permit the correct development of the conceptuses over a long period of time. The progeny of an injected cell, which forms a clone, was analyzed after implantation on embryonic day 5.5. This experiment has shown that the domain expressing the Cerl-GFP transgene is composed of a few clonally related cells (Figure 10.4, B and C Perea-Gomez and S.M.M., Piotrowska-Nitsche, Gray, Collignon and Zernicka-Goetz, unpublished observations). This indicates that the AVE has a polyclonal origin, that it is derived from...

CD Studies of Longer Peptoid Helices Containing aChiral Aromatic Side Chains

In general, peptoid helical structure is thermally stable in both organic solvent and in aqueous solution 4, 44, 47 . Complete thermal unfolding of peptoid helices has so far not been observed 44, 47 . However, studies of several different peptoid oligomers have shown a diminishing CD signal with increasing temperature, with spectra passing through one or more isodichroic points. These observations indicate that temperature can be used to induce partial melting of repeating peptoid secondary structure. Although it was reported that a water-soluble peptoid 30mer underwent a cooperative and reversible unfolding transition due to combined thermal and pH effects 4 , we found with further studies of this molecule that the observed phase transition was complicated by a solubility-to-insolubility, LCST-type phase transition (unpublished results).

Research on Direct Effects of PIs on Bees

A number of studies have now been published describing the effects of PI ingestion on adult worker honeybees in terms of toxicity and changes in bee gut protease activity levels.14-17 There has also been some unpublished work carried out on the effects of PI ingestion on olfactory learning behavior, which is a significant component of foraging behavior in adult bees.18,19 However there

Leaf Structure and Function

Leaf structural differences may directly or indirectly affect photosynthetic capacity (Nobel et al., 1975 Carpenter and Smith, 1981 Koike, 1988 Terashima, 1989). Alder (Alnus hirsuta) leaves that emerge at the beginning of autumn have two layers of epidermis, while leaves emerging earlier have only one layer. The two layers help to reduce the strong PPFD at low temperature in autumn (Koike, unpublished data). By contrast, late-successional species usually flush leaves only once, in the beginning of the growing season, and they have thinner leaves without any well-developed epidermis at the adaxial side of a leaf. These tree species with thinner leaves usually have high susceptibility to photoinhibition as compared with early-successional species (Kitao et al., 2000).

Roots and the Architecture of Root Systems

Generic Photo Shoot

Fig. 2.1 Diagrammatic representation of generic (a) dicotyledonous, and (b) monocotyledonous plants with commonly used root nomenclature. (Redrawn from unpublished work of the International Society for Root Research.) Fig. 2.1 Diagrammatic representation of generic (a) dicotyledonous, and (b) monocotyledonous plants with commonly used root nomenclature. (Redrawn from unpublished work of the International Society for Root Research.)

Rat Secretin Receptor

Mutations, deletions, or exchange of parts of the secretin receptor by the corresponding domain of the parent receptors VIP, PACAP, or glucagon have been performed to delineate the areas necessary for an appropriate structure. It is however difficult to make the difference between receptors modifications leading to an impaired structure from those that modulate ligand binding without major change in the general structure. Mutations in the N-EC domain of Asp residue 49 into Arg (but not by an uncharged polar or hydrophobic residue), of the Arg residue 83 by an Asp or by a Leu residue led to almost inactive receptors. Similarly, mutations in the EC 1 domain of Asp 174 into Ala but not Asn, or mutation in the EC2 domain of Arg 255 by an Asp or a Glu residue also severely impaired ligand recognition 12 . No functional receptor could be obtained after replacement of N-EC or EC3 by the corresponding sequences of the glucagon receptor 13 . At the opposite, deletion of the highly charged...

Well Differentiated Thyroid Carcinoma

In parallel with the lack of concrete evidence that RAI therapy has definite benefits in pedi-atric WDTC, there is lack of data regarding the formal demonstration of survival benefits and prevention of morbidity with THST 87 . On the other hand, THST is associated with fewer side effects (both short- and long-term) than RAI therapy, and has been proven beneficial in adults 86,89 unpublished data from the National Thyroid Cancer Treatment Cooperative Study Registry, Houston, TX . Furthermore, there is a strong biological basis for its long-term use in the context of WDTC management

Dispersion Of Dosage Forms In The Stomach

There have been relatively fewer studies of the behaviour of soft gelatin capsules in man. Pilot studies indicate that the time of disintegration of soft gelatin capsule formulations is highly variable, particularly if the formulations are given without food. The capsules were predominantly broken up as they entered the pylorus immediately prior to leaving the stomach (unpublished data). Armstrong and co-workers69 have compared the dispersion of oils from soft gelatin capsules in man and rabbits using x-ray techniques and gamma scintigraphy. Soft gelatin capsules were filled with iodinated cotton seed oil (Lipiodol) for x-ray studies, or iodine-123 labelled ethyl oleate for gamma camera studies in humans. In x-ray studies of rabbits, disintegration of the capsule began after 2 to 3 minutes, swelling into a more isometric shape. This behaviour was observable in vitro and was associated with the breakdown of the capsule at the sealing line. Subsequently it was difficult to assess...

Type Dependent Regulation of ACs

Tive for types I, II, III, IV, and VI, negative for type V C. Day, J. Codina, and L. Birnbaumer, unpublished). Not known is of course the relative abundance of each protein. Type VI may predominate because of the marked inhibition that somatostatin receptors can elicit, and the type VI enzyme is exquisitely sensitive to inhibition by Gi (see below). On the other hand, in view of the lesser extent of inhibition seen under basal conditions, when compared to Gs-stimu-lated activities, it is likely that the enzyme that is seen to be inhibited under basal conditions is the type I enzyme. This enzyme is known to have a relatively high basal activity as compared to that of the type VI enzyme, which has low basal activity.

Bioinformatics Terminology Methodology and Applications to RNA MTases

Gene Subfunctionalization

Guided by bioinformatics, bacterial and archaeal homologues of Trm6a have been cloned and characterized functionally. As expected, the bacterial protein cloned from Thermus thermophilus genomic DNA (termed TrmI) turned out to be a homotetrameric, site-specific AdoMet-dependent MTase, able to methylate m*A58 in tRNA in the absence of any other protein (Droog-mans et al. 2003). Surprisingly, the archaeal ortholog of TrmI (cloned from Pyrococcus abyssi) was found to exhibit not only the m1A58, but also the m*A57 specificity (Droogmans and coworkers, unpublished data). The latter finding was quite surprising a typical non-homology approach aimed at the identification of an Archaea-specific RNA modification enzyme would suggest searches for an Archaea-specific gene. This study suggested that a function specific to a given phylogenetic lineage could be in fact conferred by a moonlighting protein (Jeffery 1999),which developed a novel activity, while maintaining the original one.

Methods applications and concepts of metabolite profiling Secondary metabolism

Glufosinate Metabolism

In a separate study on biotic stress, Broeckling and colleagues reported a major reprogramming of carbon flow from primary towards secondary saponin metabolism in response to methyl jasmonate elicitation in Medicago truncatula 39, 72 . Based on metabolic profiling of both primary and secondary metabolism, a mechanistic response model was proposed and is presented in Figure 1, which involves a major reprogramming of carbon from primary metabolism towards secondary metabolism (i.e., triterpene saponins). The response includes increased levels of serine glycine threonie metabolism which is believed to result in increased levels of branched chain amino acids suggesting increased hydroxylmethylgluturate (HMG) levels. The increased levels of the polyamine beta-alanine and putrescine imply increased levels of the HMG-CoA ester which serves as the source of carbon for triterpene saponin and sterol production. However, no increase in sterol accumulation was observed supporting carbon flow...

Prophylaxis in Fellow Eye of Primary Retinal Detachment What Not to Do and What to Do

Found that eyes with pre-existing retinal breaks do not have any increased risk of retinal detachment following PVD. The same was true of eyes with pre-existing senile retinoschisis (N.E. Byer, unpublished observations). In the group of eyes with pre-existing lattice degeneration the onset of sudden PVD did lead to new retinal tears in 24 (N.E. Byer, unpublished observations). In 76 of the eyes with lattice degeneration, the occurrence of a PVD did not produce any complication of any kind (N.E. Byer, unpublished observations). However, in 50 of the lattice-eyes the new tears appeared in normal -appearing retinal areas and, thus, could not have been pretreated (using the prophylactic method) because they would not have been detected.

Embryo and Endosperm Development

The maize in vitro zygote is metabolically highly active. Newly formed cell wall material can be detected as early as 30 sec after in vitro gamete fusion 26 . After IVF, karyogamy was observed as early as 35 min 27 to 45 min 34 in egg cells and 1 h in central cells 29 . It is completed both in the egg and in the central cell within 2 h after fertilisation in vitro (HAF). The time course of karyogamy was determined by using isolated, DAPI-stained nuclei of fertilised egg and central cells 29 . Two types of karyogamy were observed in in vitro fertilised central cells. The sperm nucleus fuses either with one of the two polar nuclei or with the secondary nucleus which can be formed prior to pollination and fertilisation. This was also found in maize 35 . In vitro produced maize zygotes 17, 23, 26, 27, 32, 33 and in vitro produced wheat zygotes 28 divide in culture. Depending on culture conditions, in maize it occurs as early as 29 HAF (E. Kranz, unpublished data), but generally 42-46 HAF...

Nitric Oxide Synthesis In Plants

DNICs have been extensively studied in mammals and measured under various different physiological conditions 38-40 . Detection of DNICs in plants by EPR faces the problem that there are relatively high manganese levels in plant tissues. One of the hyperfine splitting components of EPR signal from manganese complexes mask the 2.03 EPR signal of DNICs 36, Graziano, Simontacchi, Puntarulo and Lamattina, unpublished results . Therefore, the detection limit for DNICs by EPR in plants is higher than in animals. Formation of DNICs from endogenous-free iron and thiol-containing ligands was first demonstrated in isolated leaves from bean and China rose treated with exogenous gaseous NO 41 . Recently, it was shown that treatment with high concentrations of nitrite, a condition that stimulates NO production, induced the formation of DNICs in isolated China rose leaves 23 . Incubation of the leaves with nitrate, a competitive inhibitor of the NR-dependent NO-synthesis reaction, resulted in...

The Hybridsite Model for Elongation

In functional studies, the P-site is operationally defined as the site where an acy-lated-tRNA can react with puromycin, in contrast with an A-site location where it is for all intense and purposes puromycin unreactive. The observation that an acylated tRNA can in fact undergo the puromycin reaction 26 does not seriously challenge this definition, since the latter reaction differs qualitatively from that of the P-site reaction in that it is extremely slow. For example, under conditions of 6 mM Mg2+ and in the presence of polyamines, the A-site puromycin reaction proceeds 200 times slower than that in the P-site (A. Potapov, C. Spahn and K. H. Nierhaus, unpublished results). Note that the hybrid-site model uncouples the functional definition of the P-site from the structural one, because this model locates the peptidyl residue at the P-site of the PTF center (on the 50S subunit) after the peptide-bond formation (because the peptidyl-tRNA is at the A P-site), yet the peptidyl residue is...

Inducible ARKO and Arko Tramp Mice

These two models suggested the time-specific role of AR in the development of prostate. What is the role of AR between embryonic and adolescence The time-specific role of AR in the normal prostate development can also be elucidated by the study of the inducible ARKO mice. The method uses an interferon-responsive promoter to control the expression of Cre recombinase. Here, Cre was used to delete a segment of the AR gene flanked by loxP recombinase recognition sites. Deletion was complete in liver and nearly complete in lymphocytes within a few days, whereas partial deletion was obtained in other tissues. Crossing the flox-AR female mice with Mx-1-inducible Cre male mice17 will generate interferon-inducible ARKO mice. Our preliminary data (Xu and Chang, unpublished data), showed the efficiency of deletion of AR gene in prostate tissue was about 20 using the intraperitoneal injection of pI-pC. Some modifications can be made to improve the percentage of...

Immobilization of Proteoglycans Glycosaminoclycan Chains and Oligosaccharides

Binding Dextran Surface Cells

Identification of zero background surfaces. Ligate is added in the buffer chosen for binding assays at the highest concentration likely to be used in these assays. The most commonly used detergent is Tween-20. In 0.5 (v v) Tween-20 no background binding is observed with crude lysates of tissues or cells on streptavidin-derivatized IAsys aminosilane surfaces (Wain-wright, G. and Fernig, D.G., unpublished observations). Changing the charge of the surface at physiological pH, e.g., negative charge of carboxymethyl dextran, is accomplished by repeated activation of the charged groups of the surface. An elegant example is provided in ref. (8). Fig. 2. Identification of zero background surfaces. Ligate is added in the buffer chosen for binding assays at the highest concentration likely to be used in these assays. The most commonly used detergent is Tween-20. In 0.5 (v v) Tween-20 no background binding is observed with crude lysates of tissues or cells on streptavidin-derivatized...

David G Fernig 1 Introduction

The essence of experiments using optical biosensors is that one partner of a molecular interaction is immobilized on the surface and the interaction of the other partner(s) is then followed in real time by adding them to the liquid phase. It is usual to refer to the immobilized molecule as the immobilized ligand or ligand and the soluble partner(s) as the soluble ligate(s) or ligate(s). The signal obtained from proteins and nucleic acids depends solely on the amount of material at the surface. Hence with these macromolecules, the optical biosensor acts as a very sensitive mass sensor. However, glycosaminoglycans have a low refractive index, which may vary with the degree of sulfation (Fernig, D. G., unpublished). Therefore, compared to proteins, glycosami-

Clinical Features

Good, Holland and Taylor (unpublished observations) have found no association between convulsions and a raised Toxocara titre in a community study of over 100 000 schoolchildren but have found an association between convulsions and toxocaral eye disease in a case-control study. In a case-control investigation of human Toxocara infection of the central nervous system, Magnaval et al. (1997) failed to find an association between case status and clinical signs but did record a signification association between case

PEG Prodrugs of Amines

Analogous conjugates to the DNR series were prepared for doxorubicin (Enzon Pharmaceuticals, Inc. unpublished results), and tested in an MX-1 xenograft mouse model that is very responsive to DOX. Unexpectedly, PEG-conjugated derivatives produced linker profiles (Table 9) different from than that observed for DNR in the M109 mouse model. In the DOX case the aromatic

Other Afferent Systems Acetylcholine Norepinephrine and Histamine

In rats, chronic administration of the alpha-adrenergic agonist, clonidine, shortens free-running period in both constant light and constant darkness (Rosenwasser, 1996), but the magnitude of this effect increases as a function of increasing illumination, which itself normally lengthens circadian period (Dwyer and Rosenwasser, 2000b). Similarly, in hamsters, chronic clonidine administration blunts the phase shifting effects of brief light pulses (Dwyer and Rosenwasser, 2000b), and acute clonidine treatment evokes circadian phase shifts according to the photic-type PRC (Rosenwasser, Vogt, and Pellowski, 1995). Taken together, these results suggest that alpha-adrenergic mechanisms affect the circadian pacemaker directly, and also modulate photic input to the pacemaker. While the anatomical loci of these effects are unknown, chemotoxic lesions of noradrenergic terminals using DSP-4 has been reported to block the phase-shifting effect of clonidine, suggesting that presynaptic alpha-2...

Recommendations for the Development of Written Discharge Health Information

Unpublished research by Mellow and Christian (2005) indicates that discharge materials produced for inmates are presently not tested to ensure the appropriate readability level. In a nationwide sample of reentry guides analyzed by Mellow and Christian, no reentry guides were written at the fourth or fifth grade level and the majority of the guides were written at the high school or college level. Information written above an inmate's level increases the frustration they already experience when returning to the community.

Significance of Chlorophyll Breakdown

It is understandable that in an alga such as Chlorella, Chl breakdown is less elaborated enforced Chl breakdown in darkness and under heterotrophic conditions proceeds only to the level of macrocycle cleavage and the first product, RCC, is excreted into the surrounding medium (Oshio and Hase, 1969 Engel et al., 1991). In the evolution of Chl catabolism, further photodynamic inactivation of RCC by RCCR probably coincides with the transition to terrestrial life and to intracellular disposal of catabolites. Indeed, the activity of RCCR has been detected in several taxa of pteridophytes including primitive ferns (Matile group, unpublished results). Nevertheless, alternative mechanisms of Chl breakdown almost certainly play a role at stages of development other than senescence. Thus, turnover of Chl in developing and mature leaves (Stobart and Hendry, 1984) is not associated with the accumulation of NCCs (Matile group, unpublished results) suggesting that the PaO pathway is not involved....

Special Education Costs

As with the cost of medical care beyond age 5 years, an estimate of the special education costs due to premature birth was made on the basis of the incremental special education costs specific to four developmental disabilities that have a higher prevalence among those born pre-term MR, CP, VI, and HL. It is noted that the increased use of infertility treatment may lead to special education costs due to its effect on preterm birth via multiple gestations. To the extent that preterm birth associated with multiple gestations leads to the four disabilities that are included in these estimates, such costs are included. The estimates reflect the receipt of services from ages 3 to 18 years, discounted back to the year of birth at a 3 percent rate. The estimates were based on data from CDC (2004c) and unpublished tabulations from MADDSP, as described above. The rate of receipt of special education services by developmental disability was based on the distribution from MADDSP. Allocation...

Role Of The Smad Pathway In Furin Gene Regulation

In a recent study, evidence was provided for the central role of Smads in the transcriptional activation of the TGF -inducible fur promoter 23 . As described by Ayoubi et al., three alternative promoters direct fur expression 37 . Promoters P1A and P1B are very GC-rich and contain several SP1 sites, so they have been proposed to be involved in maintaining the housekeeping levels of this enzyme. Promoter P1, on the other hand, is of the inducible type. It has both TATA and CCAAT elements in the proximal promoter region, and was reported to be trans-activated by the transcription factor C EBP, a feature relevant to its relatively high levels of expression in hepatocytes. Using HepG2 cells transfected with various furin promoter constructs, it was observed that among the three furin promoters, the P1 promoter was the strongest and most sensitive to TGF 1 and that Smad2 together with winged helix transcription factor FAST (FoxH1) participated in this transactivation 23 . It was also found...

Review Of Developmental Research On Prospective Memory

On the other hand, it would be incorrect to conclude that prospective memory is largely age invariant and that adjusting task difficulty in the developmental studies of prospective memory will always eliminate the age effects. For example, in two unpublished studies we modified the method developed by Kvavilashvili et al.

Translational Regulation

Regulation (46 Hanfrey, C., Elliott, K. A., Franceschetti, M., Mayer, M. J., Illingworth, C., and Michael, A. J., unpublished data). The plant tiny uORF small uORF arrangement and the mammalian MAGDIS uORF are so different in sequence, size, and position on their respective mRNA 5' leaders, that they may represent an intriguing case of convergent evolution.

The E coli RNA degradosome

The association of RNase E and PNPase in a complex could provide a direct physical link for their cooperation in degradation. Indeed, there is in vivo evidence for coordination between endonucleolytic cleavage at the 5'-end by RNase E and attack at the 3'-end by PNPase 19, 21, 85 . RNA I, a small 108 nucleotide molecule, is a repressor of ColE1 plasmid replication with a short lifetime similar to that of an mRNA. Although the primary transcript with a 5'-triphosphate end and 3'-RNA stem-loop structure is resistant to PNPase attack, removal of five nucleotides from the 5'-end by RNase E triggers the decay of the RNA I-5 intermediate in a pathway that involves 3'-polyadenylation and degradation by PNPase. It has been suggested that poly(A) polymerase and PNPase might be recruited to RNA I-5 by their interaction with RNase E in the degradosome. Indeed, with the rne131 mutant (see below), which disrupts the RNA degradosome, there is significant stabilization of the RNA I-5 intermediate...

In Vivo Prostate Tumor Targeting And Imaging

The potential toxic effects of semiconductor QDs have recently become a topic of considerable importance and discussion. Indeed, in vivo toxicity is likely a key factor in determining whether QD imaging probes would be approved by regulatory agencies for human clinical use. Recent work by Derfus et al. indicates that CdSe QDs are highly toxic to cultured cells under UV illumination for extended periods of time (60). This is not surprising, because the energy of UV-irradiation is close to that of covalent chemical bond and dissolves the semiconductor particles in a process known as photolysis, which releases toxic cadmium ions into the culture medium. In the absence of UV irradiation, QDs with a stable polymer coating have been found to be essentially nontoxic to cells and animals, for example no effect on cell division or ATP production (D. Stuart, X. Gao, and S. Nie, unpublished data). It has also been reported that the polymer-coated QDs could be toxic if significant aggregates are...

Obtaining Cytological Histological Diagnosis

Although historically cutting needles have not been used in the neck due to the theoretical risk of damage to other tissues, when used with US guidance the dangers are extremely small. As such this is the routine method in our hospital for impalpable nodules, where there has been a failed previous FNA or for recurrent disease, that is, for the most difficult cases. There have been no major complications. A double spring action needle is preferred (the inner stylette and the outer cutting sheath are both fired by spring action) as the size of the sample obtained is maximized. The needle is fired on the touch of a button on the handle for a fixed distance (either 2.2 or 1.1 cm). Under local anesthetic and ultrasound vision the needle is advanced to the target nodule and the path of the needle once fired can be considered to hit the target and miss important structures. Cutting needles significantly improve the quality of the sample obtained for the cytologist, reducing the...

RRP after radiotherapy

One of the major concerns with salvage surgery has been the high reported rate of complications, including the risk of rectal injury and incontinence, as well as high rates of positive surgical margins.94 In the Mayo Clinic series mentioned above, the rates of rectal injury, bladder neck contracture and urinary incontinence were 6 , 21 and 50 , respectively.93 In fact, some have advocated radical cystoprostatectomy in this setting because of these concerns.95 In a more recent review from MSKCC, rectal injury is now uncommon, but incontinence remains problematic.96 One potential advance is that of preoperative endorectal MRI evaluation urethral length measured on coronal MRI sections has been correlated with postoperative continence (unpublished). Patient selection for salvage RRP remains paramount despite the reduction in morbidity and complications, the incidence of advanced disease on final pathologic examination of the prostate and pelvic lymph nodes remains high.94 Patients must...

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