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Kindle Money Mastery

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Chromosomal abnormalities in cancer

Chromosomal and DNA ploidy is an important parameter which has served as a marker of prognosis in breast cancer (Hedley et al., 1987 Clark et al., 1989 Ferno et al., 1992 Grant et al., 1992 Wenger et al., 1993), as well as in other forms of cancer such as pancreatic adenocarcinoma (Porschen et al., 1993), melanoma (Karlsson et al., 1993), endometrial cancer (Rosenberg et al., 1989), and gastric leiomyosarcoma (Suzuki and Sugihira, 1993). An image cytometric study carried out in the authors' laboratory on breast cancer aspirate cells has also revealed a highly significant relationship between ploidy and prognosis (see Table 2) (G. V. Sherbet et al., unpublished data). Nevertheless, it should be noted that a dissenting view has been expressed with regard to the significance of aneuploidy as a prognostic indicator (Lanigan et al., 1992 Lipponen et al., 1992). Aneuploidy may be a consequence of cells entering the S-phase of the cell cycle prematurely. This can be inferred from the close...

Nutritional supplement

The redox potential of Cr(pic)3 has recently been reported 91 . The compound is considerably harder to reduce than previous estimates suggested. In DMF, the Cr(II) Cr(III) couple has an E1 2 of -1.23 V vs. normal hydrogen electrode, while the dinuclear compound Cr2( -OH)4(pic)4 is harder to reduce (E 2 -1.45 V) 91 . No features are observed at positive potentials until solvent decomposition occurs for either compound. The potential for the reduction of Cr(pic)3 is slightly shifted in the positive direction in water (A. Pickering and J. Vincent, unpublished results). The implications of this for the toxicology of Cr(pic)3 will be discussed in later chapters.

Cloning the Mutated Genes

It is possible that no insert appears linked to the phenotype in our screen, we found that about one quarter of the mutants recovered were not linked to a detectable insertion. In addition, it is important to note that the identification of an insertion initially linked to the phenotype is not proof that the identified insert is tightly linked to the mutation it is merely a way to either identify the insertion that is a candidate for causing the mutation or to conclude that the mutation is not linked to any insert if no insert meets the criteria. This is because recombination rates in the male germline are much lower than in the female germline (Johnson, personal communication Amsterdam, unpublished data) thus, an insert inherited from an F1 male that is merely on the same chromosome as a noninsertional mutation will often meet the previously mentioned criteria. The mutation and the insert will not have segregated in the F2 generation, and because the mutant F3 embryos must inherit...

The foundations of goalbased thinking

When this approach to moral thinking is brought to the ethics of research on humans, we are, in effect, asking what use the research results will be to medicine. This concern with the medical or scientific value of research also obliges us to think about two other related issues. These are, first, that the research should be designed in such a way that the goals are achieved satisfactorily, and second, that when they are achieved, they should be disseminated. In a sense, these are technical questions, not ethical ones. But they have serious ethical implications. Badly designed research gives rise to inaccurate results which mislead and are potentially dangerous to patients. Unpublished results, however accurate, help no one except the researcher. The rest of this chapter will address these three areas of moral concern which fall under the heading of goal-based issues.

Somatosensory Cortex In Chimpanzees And Humans

Electrical stimulation of the dorsal nerve of the penis and recordings of evoked potentials with surface electrodes placed the representation of the penis more laterally with hip and upper leg (Bradley et al., 1998). This seems to be different from monkeys where the posterior leg, genitals, and tail appear to be located in the medial wall, just ventral to the foot in areas 3b and 1 (Nelson et al., 1980). However, in a recent, yet unpublished, reinvestigation in macaques, we found the genitals to be represented just lateral to the foot and leg in area 3b. The use of electrical stimulation of the nerve in humans resulted in an overestimation of the proportional size of the representation of the penis compared to other body structures (Bradley et al., 1998).

Functional Implications

Orexin fibers innervate the paraventricular and arcuate hypothalamic nuclei, as well as the LHA itself. These hypothalamic nuclei have all been classically implicated in neuroendocrine, autonomic, and metabolic regulation. Orexin receptor mRNA, particularly OX2R mRNA, is expressed in these regions. Arcuate neurons in particular, some of which express NPY and are thought to be involved in the control of feeding and body weight homeostasis, are excited by orexin administration (70,71). Similarly, MCH neurons in the lateral hypothalamus are activated in response to application of orexins (47,72). Several hindbrain regions involved in central autonomic control and metabolic regulation are innervated by orexin neurons and include the dorsal motor nucleus of the vagus, the nucleus of the solitary tract, the nucleus ambiguous, and the rostral ventral lateral medulla. These regions express orexin receptor mRNA, and several studies have shown direct excitatory and in some cases inhibitory...

Mistargeting of RNA to ER Subdomains Affects Protein Localization

The preceding description clearly demonstrates that at least for one protein, the 10-kDa 8-zein, the localization of its RNA has a profound impact on where the protein is deposited in the cell. The converse experiment to support the relationship between RNA localization and protein localization is to mistarget an RNA from the cisternal ER to the PB-ER. The sunflower seed 2S albumin (SSA) is stored in the protein storage vacuole in both sunflower and transgenic rice. Closer examination showed that SSA protein is localized on the periphery of PB-II (Washida et al. unpublished) much like the 26-kDa globulin (Krishnan et al. 1986). Consistent with this location in the storage vacuole, its RNA is observed on the cisternal ER much like that seen for glutelin RNAs. Targeting of SSA RNA to the cisternal ER suggests the presence of cis elements functionally equivalent to those present in glutelin RNA (Hamada et al. 2003b) The SSA RNA was modified to contain the 5' coding sequences of the...

Callosal Axons and Their Development

Third, each area is callosally connected to its own characteristic set of other areas. For example, the 17 18 region of the cat is connected to the contralateral 17 18 region as well as to peristriate areas such as 19 and 21a, the suprasylvian visual areas (Segraves and Rosenquist, 1982), and, in addition, to the insular cortex and to the claustrum (unpublished). The connections are usually reciprocal, and in general, areas that are callosally connected are also connected intrahemispherically (Figure 1.3). This suggests that areas in opposite hemispheres might communicate through a number of alternative intrahemispheric and interhemispheric routes, which might become differentially active in different functional conditions.

Collecting the results of research

Different from its standard comparator, he would be less successful. The implication is that important negative results have not seen the light of day. Easterbrook concludes 'These findings suggest that conclusions based only on a review of published data should be interpreted cautiously, especially for observational studies. Improved strategies are needed to identify the results of unpublished as well as published studies.' In 1997 between 50 and 100 reputable medical journals called an amnesty for unpublished trials. Investigators with unreported trial data were invited to register their trials by completing an unreported trial registration form. The aim, said Roberts (1998), 'was to tap the silent subterranean pool of unpublished research and by bringing these data to the surface, to increase the power of systematic reviews and reduce the effects of publication bias'. At the time of writing, which was a year after the amnesty had been issued, Roberts reported that only 165 such...

Transduction Efficacy

Adenoviral transduction efficiencies of the first generation adenoviruses in different animal species six days after the gene transfer. LacZ immunostainings (brown) demonstrate the distribution of the nuclear-localised transgene. (a) In rabbits interstitial cells such as fibroblasts and also skeletal myocytes are transduced by intramuscular injections of adenoviruses (total dose 1 x 1011 viral particles (vp)). (b) Cardiac myocytes in the pig myocardium are efficiently transduced by intramyocardial injections of adenoviruses (total dose 4 x 1011 vp) (Markkanen M, unpublished 2007). (c) Almost no transgene expression can be detected in mouse skeletal muscle after intramuscular injections of adenoviruses (2 x 1011 vp). (d) In mouse skeletal muscle, transgene expression is only visible in the damaged cells around the needle track Figure 1. Adenoviral transduction efficiencies of the first generation adenoviruses in different animal species six days after the gene transfer. LacZ...

Duration Of Gene Expression 31 Gene Expression Profile

The expression profile of the first generation adenovirus (AdV) compared to gutless adenoviruses (HCAd) and AAV in vivo. Measurements of the secreted alkaline phosphatase (SEAP) activity from rabbit sera at several time points after adenoviral transduction display adenoviral transgene expression kinetics (Korpisalo P, unpublished). Transgene expression begins shortly after the transduction (day 0) but is also rapidly shut-down after the peak expression has been reached (day 6). A second administration at day 14 cannot boost the expression. With gutless AdVs gene expression (kinetics estimated on the basis of ultrasound perfusion data) is lengthened compared to the first generation AdVs but the expression still fades since the transgene does not integrate into the host genome (Korpisalo P, unpublished). However, With long-term expression vectors, such as AAV or lentiviruses a real, long-lasting expression can be achieved (AAV gene expression profile estimated on the basis of...

Activation of TLR9 proceeds in a multistep process

Ferent DNA sequences and structures Stimulatory and non-stimulatory sequences bind TLR9 74 (and unpublished observations). This is in accordance with the notion that non-stimulatory DNA can competitively block TLR9 activation by stimulatory DNA sequences. In recent years potent TLR9 inhibiting sequences have been worked out. These inhibitory ODN are commonly about 15 bases long carry the phosphorothioate modification and contain a pyrimidine-rich triplet preferably CCT, which is positioned with a spacer 5' to a GGG sequence 124, 125 . Inhibitory ODN block the immunostimulatory effects of microbial DNA and activating CpG ODN. Most likely inhibitory ODN act as competitive antagonists at the level of TLR9, but this has to be formally proven.

Molecular Genetic Studies

Is dependent of genetic background (our unpublished observation), these observations suggest altered energy homeostasis in orexin-deficient narcolepsy. On the other hand, acute pharmacological blockade of orexin-1 receptor (OXjR) results in hypophagia and decreased body weight (10). This paradoxical phenomenon might suggest that receptor blockade is necessary for developing obesity. In fact, we observed that OX1R knockout mice do not develop obesity (our unpublished observation). It is also possible that chronic disruption of orexin signaling by gene targeting and acute or semichronic blockade of orexin receptor might have different impacts on the orexin-mediated feeding pathway and an orexin-mediated increase in metabolic rate.

Orexin And Other Feeding Behaviorrelated Functions

Orexin might also participate in other feeding-related functions. Tuberomammillary nucleus (TMN) histaminergic neurons were shown to be activated by mastication owing to a signal originating from oral proprioceptors and initiated by chewing (28). Reciprocally, histaminergic neurons densely innervate the mesencephalic trigeminal sensory nucleus. This pathway was shown to be involved in the regulation of mastication. Because orexin neurons densely innervate the mesencephalic trigeminal sensory nucleus and the TMN, and orexin receptor mRNAs exist in these nuclei, it might be possible that orexin neurons are also involved in the regulations of mastication. In fact, we usually observed a chewing-like movement of the mandible in rats after orexin was administered centrally (our unpublished observation).

Host Biology and Distribution

The Microchiroptera are physically smaller and generally insectivorous animals that navigate by echolocation (Hall and Richards 2000), of which 65 species have been identified in Australia. One species, Saccolaimusflaviventris, has been shown to be infected with a different strain of ABLV to that of the Pteropus strain (Gould et al. 2002), and there is an unpublished observation of ABLV infection in other Microchiropteran species (Mackenzie et al. 2003).

Introduction 411 Overview

The next step was investigating the properties of tubulin and MTs in depth since once experimentally implicated in information manipulation, it was not unreasonable to ask whether these protein structures do indeed work as biological and perhaps quantum binary digits (biobits or bioqubits). The dielectric constant and electric dipole moment of tubulin were chosen for closer study since these featured prominently in information manipulation schemes by naturally occurring or fabricated MT networks. In order to check the validity of previous computer simulations first done by the group of Jack Tuszynski 16 and later replicated by our group (giving virtually identical results 87 ), an experimental determination of the dielectric constant of tubulin and MTs was needed. This was undertaken in the high-frequency (optical) region (using refractometry and surface plasmon resonance) 87 and the low-frequency region (using dielectric spectroscopy - previously unpublished data and reported here in...

Offspring production and recruitment

Numerous studies have repeated Lack's observation of a decline of clutch size with laying date black-capped chickadees (Kluyver 1961 S. M. Ramsay unpublished data) marsh tits (Smith 1993) blue tits (Nilsson and Svensson 1993). The box-nesting habits of Eurasian tits have allowed researchers to manipulate nesting conditions, such as inducing females to delay laying these experiments have resulted in reduced clutch sizes in manipulated females (great tits, Verhulst and Tinbergen 1991, Barba et al. 1995 blue tits, Sanz 1999). The effects of delayed laying were population-specific in one study of great tits, with the quality of individual territories and vegetation-related food availability between the populations likely causing the difference (Verhulst and Tinbergen 1991). models to test whether future survival is affected by hatch date. Other North American Parids which exhibit higher philopatry may be more suited to such studies. Regardless, there is cause to suggest that early nesting...

Cellular phenotype in primary screening

The value of an image-based cell death assay is apparent when considering that some drug candidates might cause cytostasis, only reducing cell proliferation without actually causing cells to die. Commonly used single-readout assays, such as annexin staining or caspase activation, fail to detect cytostasis if not coupled with a means of measuring absolute cell number. A simple comparison of cell numbers at the beginning and end of an assay period, such as that afforded through image analysis, is less likely to yield false negatives arising from candidates that act through annexin (1,2) or caspase-independent pathways (reviewed in refs. 3 and 4). The importance of simply observing cell cultures cannot be underestimated. With various cell compound combinations, Automated Cell scientists have detected dividing cells that stained positive for either PI or annexin (unpublished observations and reviewed in ref. 5). The automated nature of the ACI platform allows for screening of millions of...

Nonconventional Antisense in Zebrafish for Functional Genomics Applications

(Urtishak et al., 2003), but they also appear to have a reduced perdurance when compared to MOs. For example, gripNAs do not appear to effectively target the function of the nacre gene (Pickart and Ekker, unpublished observations), whose Chemically, MOs are inherently very stable molecules. A highly pure preparation maintains its activity over several years (Ekker, unpublished observations). For greatest preservation of activity, lyophilization is effective but not very convenient. Aqueous solutions can be stored at 80 C for long-term storage where only limited access is required. Storage of aqueous solutions for ready access is more problematic, however. Multiple freeze-thaw cycles of aliquots stored at 20 C can reduce efficacy, presumably through the formation of complex interoligonucleotide interactions. Sometimes, this effect can be partially reversed by heating the affected solution. To reduce this effect, storage of solutions at 4 C is also viable, but this method requires...

Coronavirus S Proteins And Their Receptors

The crystal structure of the SARS-CoV RBD is consistent with this speculative possibility.78 The RBD contains two subdomains a core and an extended loop. The core is a five-stranded, anti-parallel P-sheet, with three short connecting a-helices. The loop, residues 424-494, termed the receptor-binding motif (RBM), is the only domain that contacts ACE2 directly. Although the RBD core domain is homologous with similar regions of other group 2 coronaviruses, the RBM is unique to SARS-CoV. Some evidence supports the suggestion that the RBM has been acquired from another coronavirus, perhaps a group 1 virus relative of HCoV-NL63. As indicated, HCoV-NL63 also enters cells through ACE2,34 and its extended RBD region includes a stretch of residues with weak homology to the SARS-CoV RBM (unpublished observations).

The pattern of growth in the developing skull vault

The developmental origin of the coronal suture is of particular interest in understanding mechanisms of skull growth. This bilateral, vertical suture is formed between the frontal and parietal bones it is responsible for most of the growth of the skull in the fronto-occipital (front-to-back) plane. Interspecific grafting experiments in avian embryos suggested that both frontal and parietal bones have the same origin, either from mesoderm (Noden et al 1988) or neural crest (Couly et al 1993). However, investigation of a transgenic mouse with a permanent neural crest cell lineage marker suggests that the coronal suture is formed at the interface of a neural crest-derived frontal bone and a mesoderm-derived parietal bone (X. Jiang, H. Sucov & G. Morriss-Kay, unpublished work). This juxtaposition of tissues of different origins is likely to be functionally significant.

Protein Trafficking And Posttranslation Modification Of The Vsps

Not have Golgi that can be identified by EM, they do exhibit complex trafficking of proteins (McCaffery et al., 1994), as well as Brefeldin A (BFA)-inhibited protein transport, suggesting that functional Golgi may exist (Lujan et al., 1995a). In fact, VSP trafficking was also inhibited by BFA, suggesting the possibility that the VSPs are transported and processed by Golgi or a Golgi-like organelle (Lujan et al., 1995a). This possibility is supported by the finding that VSPs using their own signal peptide are transported through the Golgi in COS cells and become surface-localized (Nash, Conrad, Kulakova, unpublished). The full transmembrane portion of the peptide is required for proper transport of VSPs to the surface (Nash and Kulakova, unpublished). Addition of a peripheral vacuole (PV) localization signal diverts the VSP to the PV suggesting that the default pathway of membrane bound molecules is to the surface unless other signals are present (Touz et al., 2003). Mutation of the Zn...

Homotopic and heterotopic callosal connectivity

Heterotopic connections between lobes were demonstrated in two instances. The right calcarine region was shown to send interhemispheric connections not only to the whole occipital cortex, but also to the posterior part of the temporal and parietal cortices, including regions proposed to be involved in specifically human functions such as the angular gyrus (Clarke et al., 1995, and unpublished results). The middle portion of the inferior temporal, the fusiform, and the parahippocampal gyri and of the hippocampal formation on the right side was shown to send relatively dense heterotopic connections to the posterior part of the superior temporal gyrus, the planum temporale, and the supramarginal and the angular gyri (corresponding to Wernicke's area) as well as weak connections to the posterior part of the inferior frontal gyrus (corresponding to Broca's area see Di Virgilio and Clarke, 1997).

Features Of The Vsp Genes

There are a number of examples of vsp genes that are highly similar throughout the entire reading frame, suggesting that the vsp gene repertoire has been expanded by duplication and divergence. Perhaps the most remarkable example is the vsp 1267 gene, which consists of two identical copies in a tail-to-tail arrangement approximately three kb apart. Another example of gene duplication is the vspG3M-B gene (Mowatt et al., 1994), also called vspA6-S1 (Yang and Adam, 1995b), which is highly similar to the vspA6 gene, except that it contains between one and two copies of the 195 bp repeat and is located on a different chromosome. Interestingly, much of the gene has been sequenced from the WB isolate (probably from Afghanistan) (Yang and Adam, 1995b) and the G3M isolate (Mowatt et al., 1994) (from Peru) and the sequences were identical. This observation suggests that the divergence is not occurring rapidly. In other cases, vsp genes are highly similar in the 5' region followed by...

Results And Discussion

Human ACE2 and CD209L were recently identified as functional receptors for SARS-CoV.2' 13 Although both are functional receptors, human ACE2 is a more efficient receptor for SARS-CoV than is CD209L. The cell lines that we identified as productively infected by SARS-CoV were assayed for the presence of ACE2. One-step RT-PCR (Qiagen) with oligonucleotide primers conserved between mouse and human ACE2 was used to amplify ACE2 RNA from 1 g of total RNA in susceptible cells derived from various species. Sense primer (ACE2-902, 5'-cttggtgatatgtggggtaga) and an antisense primer (ACE2-1548R, 5'-cgcttcatctcccaccactt) amplify a 646-base-pair fragment of ACE2 when RNA is expressed. SARS-CoV susceptible Huh7, VeroE6, HEK-293T, and MvlLu cells1 were assayed for ACE2 transcript. ACE2 RNA was detected in all of the susceptible cell lines (Figure 1A). Qualitatively, the level of ACE2 differed among the various cell lines. VeroE6 had the strongest ACE2 amplicon, whereas Mv1Lu cells showed the weakest...

Physiology of the developing retina

Retina showing labeling of the retina with the calcium indicator, fura-2, and the spread of activity, indicated by the white pixels, across the retinal surface over time. (A, J. Demas and R. O. L. Wong, unpublished B, R. O. L. Wong, unpublished.) retina showing labeling of the retina with the calcium indicator, fura-2, and the spread of activity, indicated by the white pixels, across the retinal surface over time. (A, J. Demas and R. O. L. Wong, unpublished B, R. O. L. Wong, unpublished.)

Hybrid Clustering Techniques

Figure 4.19 HAC with Ward's criterion applied to 2450 cryo-EM images of Lumbricus terrestris (earthworm) hemocyanin. (a) Appearance of a typical micrograph (b) HAC dendrogram and class averages. The classes have (from left to right) 313, 420, 310, 311, 418, 355, and 323 images. (a) From Mouche et al. (2001), reproduced with permission of Academic Press (b) kindly provided by N. Boisset (unpublished lecture material.) Figure 4.19 HAC with Ward's criterion applied to 2450 cryo-EM images of Lumbricus terrestris (earthworm) hemocyanin. (a) Appearance of a typical micrograph (b) HAC dendrogram and class averages. The classes have (from left to right) 313, 420, 310, 311, 418, 355, and 323 images. (a) From Mouche et al. (2001), reproduced with permission of Academic Press (b) kindly provided by N. Boisset (unpublished lecture material.)

Cell and molecular mechanisms underlying the patterning and formation of the optic chiasm

Midline has been identified as ephrinB2. Similar to the situation in frogs, its expression in this region is under tight temporal regulation and can only be detected during the period when the ipsilateral projection is produced (Marcus et al., 2000 L. Erskine, S. E. Williams, and C. A Mason, unpublished results). A critical challenge for the future will be to establish if this midline expression of ephrinB is both necessary and sufficient for the establishment of the ipsilateral pathway.

Types of experimental designs

On the other hand, Figure 2.12C shows a problematic case. The orbitofrontal cortex response to a 3 s presentation of an aversive visual stimulus is somewhat prolonged relative to the model, but the estimated magnitude is reasonably accurate (unpublished data). The right panel, on the other hand, shows the response in the same brain region to anticipation of viewing an aversive picture. In this case, the timing and duration of the cognitive activity are unknown - the activity could occur at the onset of the cue, throughout the anticipation epoch, or with increases proportional to the proximity of the picture. The model fits poorly in this case, illustrating the importance of knowing the onset and duration of the stimulation for linear model analyses.

Heterogeneity in sperm populations and in sperm

An additional problem derived from the division of the sperm in several structures is the distribution of energy among different compartments. Given that mitochondria are located in a specific region of the sperm, oxidative respiration and the consequent adenosine triphosphate (ATP) generation is restricted to the mid-piece of the flagellum. How then do other compartments such as the principal piece of the flagellum and the head of the sperm meet their energy needs In sea urchin sperm, a phosphocreatine shuttle has been observed to transfer high-energy phosphate from the mitochondria to the principal piece (Tombes and Shapiro, 1985) however, this system is poorly developed or entirely absent in mammalian species (Kaldis et al., 1996). The findings of a unique hexokinase type 1 (HK1) in the head and in the entire flagellum (Visconti et al., 1996) as well as the discovery of (GAPDH) in the fibrous sheath of several mammalian species (Bunch et al., 1998) suggest that glycolysis could be...

Selection Criteria For Studies

Through a combination of searching published reference lists and contacts with investigators. Unpublished prevention trials were sought through contacts with researchers, although none were found that had been completed but not published. Trials in progress are included in a later section of this chapter. Classification of evidence used in this chapter is shown in Table 6.1, which is based on the US Preventive Service Task Force Criteria28 for individual studies, and NHLBI (National Heart, Lung and Blood Institute of the NIH) evidence criteria29 for overall recommendations.

Case One Systemic Delivery of Human PDGFC or PDGFD by Adenovirus Mediated Gene Transfer in Mice

Systemic effects of both PDGF-C and PDGF-D were characterized and compared to those of PDGF-B using adenovirus-mediated gene transfer in mice (24) (Hughes et al., unpublished observations). Mice were injected with adenovirus-encoding PDGF-C (Av- In addition to the changes in liver and kidney, significant histopathologic changes were also observed in bone and lung of mice treated with Av-Pdgf-b and Av-Pdgf-d. These changes included endosteal bone proliferation in long bones and perivascular lymphoid cell infiltration in the lung. These changes were not observed in the mice treated with Av-Pdgf-c (Topouzis and Hughes, unpublished results).

Chickadee calls across the hybrid zone

That the D notes of the black-capped chickadee's chick-a-dee call are longer in duration and lower in pitch than those of the corresponding Carolina chickadee call has been known for a long time (see Chapter 13). It is therefore surprising that this vocalization has not previously received particular attention in relation to the hybrid zone between the species, apart from an unpublished thesis (Crock 1975). As Crock observed using a broad geographic approach, we found that parameters of D notes from the hybrid zone were quantitatively intermediate on average. Our observation that individual chickadees in the hybrid zone produce highly variable D notes is novel.

Phosphorylation events during capacitation

There are two important points to have in mind when analyzing the PKA pathway in sperm capacitation. First, as mentioned above, the HCO cAMP pathway is involved in rapid and slow events related with capacitation. The rapid effect of HCO and PKA activation on lipid scrambling and phosphorylation of certain protein substrates (Harrison, 2004) is on a different time scale to its effect on protein tyr phosphorylation. The former occurs within 5 min of incubation in capacitation media while the later is detectable only after a longer incubation that varies dependent on the species, being around 45 min in mouse (Visconti et al., 1995a) and more than 60 min in boar sperm (Harrison, unpublished observations). Second, PKA is a ser thr kinase that is not able to directly phosphorylate proteins on tyr residues thus, an intermediate TK must be involved in capacitation. Three possible mechanisms of interaction between PKA and protein tyr phosphorylation are as follows

How does the Proteasome Mediate the Early Steps of Neuronal Apoptosis

To determine how proteasome inhibitors prevent CGC death, we examined whether proteasomes degraded these anti-apoptotic proteins and found that proteasome inhibitors, but not caspase inhibitors, stabilized and increased the levels of IAPs (Nadia Canu, unpublished observation). This finding suggests that IAPs are targeted to proteasome for degradation during the CGC apoptosis, thus favouring caspase activity as reported in other settings (25), and likely increasing the amount of pro-apoptotic IAP substrates (see also article by Lang-Rollin and Stefanis in this volume). It should be noted however that such an increase of IAPs would not be expected to lead to inhibition of cytochrome c release, as IAPs generally function downstream of the mitochondrial checkpoint.

Parids as members of the nest web

Mountain chickadees are small-bodied, secondary-cavity-nesting birds, common in montane forests in western North America (McCallum et al. 1999). Typically, they use coniferous trees for foraging and aspen trees for nesting (Hill and Lein 1989 Aitken et al. 2002). In a long-term study on mountain chickadee populations in Northern California, nest box occupancy was highest in open pine stands (Dahlsten et al. 1992). In the Cariboo-Chilcotin region of central British Columbia, mountain chickadees are abundant, year-round residents that typically occupy stands of mixed coniferous-deciduous forest, especially where Douglas-fir (Pseudotsuga menziesii), lodgepole pine (Pinus contortus), and quaking aspen (Populus tremuloides) are present (K. Martin, unpublished data). They are the most abundant breeding Parid, with detections ranging from 0.05 to 0.24 birds ha over a 10-year period and a doubling in detection rates in the past 3 years (Figure 8.1a). The increased rate of detections was...

Components Of The Proteasome System Degraded By Caspases

Since induction of apoptosis in CGCs resulted in diminished activity of the UPS that was prevented by caspase inhibitors, we asked whether the protea-some itself was a victim of caspase-dependent attack. Interestingly, we found that the amount of the a-2-a7 subunits (10) and the P1-P7 subunits (Canu, unpublished observation) of the core 20S proteasome did not change during CGC apoptosis. By contrast, the amount of the ATPase subunit S6 of the 19S regulator of the 26S proteasome, which is necessary for ATP-dependent proteolysis by the proteasome, was reduced in a caspase-dependent manner during apoptosis of CGCs (Figure. 3). This finding is in line with results recently reported for Jurkat cells treated with etoposide or actinomycin and for PCD in flies (57, 65). In these Another mechanism contributing to impairment of proteasome activity in CGCs is the finding that histone H2A is deubiquitinated (Canu et al. unpublished observation), suggesting that depletion of free ubiquitin takes...

Interspecific interactions among smallbodied cavitynesters

Mountain chickadees occupied only one of 14 nest cavities excavated by black-capped chickadees in Alberta (Hill and Lein 1988) and we found segregation of nest use in our study (Table 8.1). Segregation of sympatric mountain and black-capped chickadees may occur as a result of interspecific behavioral dominance. In a behavioral study of sympatric populations in Utah, black-capped chickadees were frequently dominant over mountain chickadees (Minock 1972). During playback experiments in our study, black-capped chickadees exhibited higher aggression towards simulations of mountain chickadee intruders in defending nesting cavities than mountain chickadees exhibited towards simulated black-capped chickadee intruders (A.R. Norris, unpublished data). Thus, while mountain chickadees are the most abundant Parid, black-capped chickadees are the more dominant species.

Sequencealignmentbased Bacterial Phylogenies

Concatenation-based Analyses In the concatenation approach, each of the molecules in the analysis must be present in all species considered. Therefore, attempts to reconstruct the phylogeny of bacteria through sequence concatenations have employed different subsets of widely distributed proteins. Brown et al. (2001) utilized proteins conserved across species from all domains (Archaea, Bacteria and Eucarya), from which they removed those proteins producing trees that intermixed species from different domains (which was interpreted as evidence of horizontal transfer). The remaining subset of universally distributed proteins contained 14 information-processing molecules, mostly involved in translation. Other studies have deliberately focused on extensive concatenations of translational proteins (Brochier et al., 2002b), and more specifically, of ribosomal proteins (Hansmann and Martin, 2000 Wolf et al., 2001 M. P. Francino, unpublished analysis). Proteins of the translational apparatus,...

Mechanisms Of Proteasomal Inhibitioninduced Death 21 Accumulation of Rapidly Turning over Proteins

Cell cycle-related proteins and p53 are only some of the proteins that are normally rapidly degraded by the proteasome. It remains to be seen which other such proteins exist in neuronal cells, and whether their upregulation following pro-teasomal inhibition could also have deleterious consequences and lead to cell death. Potential candidates include proteins that influence the jun kinase pathway and the subsequent phosphorylation and activation of the transcription factor c-jun, a critical event in many apoptotic pathways. A prime candidate amongst these proteins is POSH, a scaffold protein that mediates activation of JNK apoptotic cascade (26, 27). POSH has a ring finger domain with putative E3 ubiquitin ligase activity. The absence of this domain leads to much higher levels of POSH upon transient transfection, suggesting that POSH is acting as a ligase for its own degradation. Consistent with this idea, levels of overexpressed POSH are regulated by proteasomes (26). Whether...

Hepatic dendritic cells

Hepatic dendritic cells are the professional antigen-presenting cells of the liver. During inflammation, dendritic cells are recruited into the liver sinusoids from where they can migrate to periportal and pericentral areas. Hepatic CD11c+ CD11b+ dendritic cells express TLRs on their cell surface and respond to LPS, peptidoglycan, poly(I C) and CpG-DNA to produce inflammatory cytokines, such as IL-12 and TNF-a and to express co-stimulatory molecules, such as CD40, CD80 and CD86 on their cell surface (Seki and Tsutsui, unpublished data). Liver CD11c+, CD8a-(myeloid) and CD11c+, CD8a+ (lymphoid-related) DC express lower TLR4 mRNA compared with their splenic counterparts 11 . Lower TLR4 expression correlates with the reduced capacity of LPS-stimulated, but not anti-CD40-stimulated liver DC to induce naive allogeneic (C3H HeJ) T cell proliferation. In contrast to LPS-stimulated splenic DC, these LPS-activated hepatic DC induce alloantigen-spe-cific T cell hyporesponsiveness in vitro and...

Accumulation of Misfolded Proteins

Consistent with the importance of protein misfolding in the setting of proteasomal inhibition, neuroblastoma cell lines stably expressing the chaperone HSP40 were partially protected against proteasomal inhibition-induced death (34). We have found a dramatic induction of the inducible form of HSP70 in a subpopulation of cultured cortical neurons following proteasomal inhibition. This presumably reflects a response to the increase in protein misfolding (35). Interestingly, dopaminergic neurons in ventral midbrain cultures fail to induce HSP70 in response to proteasomal inhibition, whereas other cells in the culture show a marked induction. This may reflect a relative inability of these specific neurons to mount this beneficial refolding response. The fact that such dopamin-ergic neurons do form inclusions and undergo apoptosis in this setting indicates that they are exposed to proteolytic stress. It appears therefore that these specific neurons may be unable to mount this beneficial...

The physiological role of ROS

The physiological importance of NADPH-induced free radical generation has been emphasized in studies of hexose monophosphate shunt activity during capacitation. The hexose monophosphate shunt is required to generate the NADPH needed by a putative, ROS-generating sperm NADPH oxidase. Involvement of hexose monophosphate shunt activity in ROS generation and sperm capacitation has been emphasized by a number of different studies. Firstly, both the tyrosine phosphorylation cascade associated with sperm capacitation and ROS generation are dependent on the presence of glucose (Aitken et al., 1998b). Secondly tyrosine phosphorylation, capacitation and ROS generation can all be blocked by the presence of a non-metabolizable glucose analogue, 2-deoxyglucose via mechanisms that can be reversed with NADPH (Aitken et al., 1998b Urner and Sakkas, 2005). Thirdly, inhibition of shunt activity with 6 amino nicotinamide also suppresses sperm capacitation via mechanisms that can be reversed by NADPH...

Process Considerations

During the genome mapping step, thousands of expressed sequences have perfect alignments with genomic segments from two different chromosomes, or from two distant portions of the same chromosome (for example, regions that are 400,000 bp apart on the same chromosome). Those expressed sequences are likely chimerical sequences, generated by mistake during EST sequencing or resulting from genomic translocations. There are well-documented fusion mRNA sequences, which resulted from translocations in tumor tissues for example, AB000267 and AB000268, AB001342 and AB001343 (Arai et al, 1997), AF060927 (Silliman et al., 1998), HSCOLPFU1 and HSCOLPFU2 (Simon et al., 1997), HSRLF (Makela et al., 1991), and AF179280 (Lagerstedt et al., 2000). The GenBank annotations of these mRNA sequences may sometimes reveal whether or not they are results of fusion events. On the other hand, those identified chimerical sequences from genomic mapping process could be used for identification of translocation...

Composition of the extracellular matrix in the physeal cartilage

An extensive network of collagen fibrils provides an 'endoskeleton' about which other matrix molecules are organized (Fig. 1). Type II collagen is the principle component of the fibrils which are thin ( 25 nm in diameter) because this is a transient cartilage, last only several days and must be easily resorbable. This contrasts to the thick fibrils found in mature articular cartilage which are usually more than 100 nm in diameter and must resist resorption and survive for the life of that individual. These fibrils lack the small proteoglycan decorin which is found on fibrils in the adjacent epiphyseal and articular cartilages (M. Alini & A. R. Poole, unpublished work). Type IX collagen is also present on the fibril during matrix assembly in a molar ratio of 5 1 (type II IX). The majority of these molecules ( 90 ) lack the basic NC4 domain that is normally found on type IX collagen in articular cartilage (Mwale et al 2000). Type XI collagen is also present within the fibril and on...

Proteasomal Inhibition And Pcd A Complicated Relationship

The reader who has gone through both the previous chapter and the current one may be somewhat baffled. Is proteasomal inhibition deleterious, as suggested by all the data presented in this chapter, or is it beneficial, as suggested by the seminal work of Sadoul et al. (66), Canu et al. (67) and other studies referenced in the preceding chapter There is no doubt that one important variable is the intensity and timing of the inhibition. As demonstrated first by Lin et al. (68), pro-survival effects occur with low concentrations, whereas pro-death effects occur with high concentrations of pharmacological proteasomal inhibitors, when applied to cultured prostate carcinoma cells. A survey of some in vivo experiments further reinforces this idea. For example, Sawada et al. (69) injected nanomolar concentrations of lactacystin in mouse substantia nigra and observed protective effects on MPP-induced neurodegeneration of dopaminergic neurons, whereas McNaught et al. (70) used micromolar...

Prior knowledge of collagenases in endochondral ossification

Earlier work has identified excessive collagenolytic activity in the hypertrophic zone of the growth plate (see Poole 1991, 1997). The content of inhibitors of collagenase (tissue inhibitors of metalloproteinases) is also reduced. Net collagen breakdown results. Collagenase has been localized in the extracellular matrix of the hypertrophic zone in the growth plates (see review Poole 1997, Blair et al 1989). Using a newly developed antibody to the collagenase-generated neoepitope in type II collagen (Billinghurst et al 1997), we have shown that collagen cleavage is present throughout the growth plate, although it is enhanced in the hypertrophic zone where net loss of type II collagen is observed (F. Mwale, I. Pidocux, E. Tchetina & A. R. Poole, unpublished results).

Endotoxininduced liver injury

Endotoxin-induced fulminant hepatitis is a common clinical complication during sepsis and accounts for a high percentage of sepsis-associated mortality. In comparison to humans, rodents are highly resistant to LPS. However, after treatment with heat-killed P. acnes, mice become highly susceptible to LPS challenge. P. acnes leads to a recruitment of macrophages and dendritic cells into the liver and a differentiation of hepatic T-helper lymphocytes into type 1 T-helper cells. The sensitization by P. acnes in the liver depends on IFN-y, IL-12 and IL-18 59-61 . TLRs are involved in both the P. acnes-priming phase and the LPS challenge phase. Indeed, hepatic granulomas, consisting of macrophages, dendritic cells and lymphocytes, were not found in MyD88-deficient mice after P. acnes administration and liver injury was blunted (Seki, unpublished data). P. acnes sensitizes towards TLR4-, but not TLR2 ligands, and upregulates the hepatic levels of TLR4 and its co-receptor MD-2 62 . Whereas...

Current studies of matrix resorption

Extracellular matrix, undergo hypertropy (synthesize type X collagen) and mineralize their extracellular matrix. This also involves resorption of the extracellular matrix at the time of mineral deposition. We have performed analyses using the reverse transcriptase (RT)-PCR and using Northern blotting on these cultures. They reveal that only MMP13 is present (W. Wu, C. Billinghurst & A. R. Poole, unpublished results). Studies have independently shown the up-regulation of MMP13 by hypertrophic chondrocytes (Gack et al 1995, Johansson et al 1997, Stahle-Backdahl et al 1997). Prior to hypertrophy there is more limited collagenase activity, which is inversely related to type II collagen and aggrecan content. This suggests a regulatory role for collagenase in matrix turnover (Mwale et al 2000). The presence of activated enzyme is markedly increased at the time of hypertrophy and coincides with an increase in type II collagen cleavage by collagenase and a rapid loss ofthis collagen (W....

Variation in rank acquisition

Analysis of rank histories of known males from our long-term data set provides a revealing picture of the factors associated with change in status (Schubert et al. 2006 K. A. Schubert, C. Kraus, D. J. Mennill et al. unpublished data). As in willow tits (Lahti et al. 1996), rank is a dynamic trait. The majority of mid- and low-status birds increased in rank over consecutive years birds reaching high rank seldom regressed (Table 9.3 Schubert et al. in press). Most birds advancing from low to high rank immediately dominated yearlings (22 29 cases), and some younger or older adult flock-mates (7 cases) (Schubert et al. 2006). Not surprisingly, males reaching high rank lived longer, on average (3.4 0.1 years, N 98) than males who never moved beyond subordinate status (1.6 0.1 years, N 99, P < 0.0001) (Schubert et al. in press). Similarly, low-ranked adult males were more likely to increase in rank than low-ranked juveniles (K. A. Schubert, C. Kraus, D. J. Mennill et al., unpublished...

Carrying Out the Reconstruction

After the screening step, which is done to verify that the tilted-particle projections follow one another in a reasonable sequence, the particle set is ready for 3D reconstruction. Initially, weighted back-projection with generalized weights was used (Radermacher et al., 1987b), but later a variety of options including a modification of SIRT (Penczek et al., 1992) and techniques based on conjugate gradients (Penczek, unpublished see SPIDER documentation) and Fourier interpolation (Lanzavecchia and Bellon, 1996 Grigorieff, 1998 Penczek, 2004) have been added (see section 4).

Vascular Maturation As A Therapeutic Target

Control over the microenvironmental dosage can be achieved through the genetic manipulation of progenitor cells ex vivo and their selection in vitro before reimplantation, as described above 41, 42 . However, while this method provides a clear proof-of-concept, it is not time- and cost-effective for a direct clinical application. To overcome this problem, our group has recently developed a FACS-based method to allow the level of expression of VEGF to be predicted in single engineered cells and the rapid purification of a population homogeneously expressing a desired level, without isolation of individual clones (Misteli et al, unpublished results). Alternatively, the controlled release of recombinant VEGF protein, achieved through its incorporation into slowly degrading polymer matrices, may allow a similar prolonged delivery of homogeneous concentrations of growth factor in vivo 43 .

Tobaccoderived Carcinogens 2231 Tobacco Components

We have investigated the metabolism of NNK in human pancreatic ductal and islet cells as well as in microsomes and cytosols isolated from histologically normal pancreatic tissue (unpublished data). All incubations of NNK with ductal cells yielded a-hydroxylated products these were formed at significantly higher rates than those observed in islet cells. Among metabolites derived from a-hydroxylation, 4-hydroxy-1-(3-pyridyl)-1-butanone (keto alcohol) was the major metabolite produced by islet cells. This compound was not detected in incubations with ductal cells. Both types of cells produced 4-hydroxy-4-(3-pyridyl)-1-butanoic acid (hydroxy acid) and 4-oxo-4-(3-pyridyl)-1-butanoic acid (keto acid). The chiral product of carbonyl reduction of NNK, NNAL was the major metabolite detected. Both cell types predominantly formed the ( )-enantiomer of NNAL. In both microsomal and cytosolic incubations with NNK, NNAL was the major metabolite formed. With regard to its formation, the microsomes...

Box 111 Repertoire sizes

Instructive to compare the quality of juvenile gargles recorded in mid-September with those sub-gargles of mid-July (Fig. 11.3b). In those two months the sub-gargle calls have developed into well-articulated gargle calls with a high degree of stereotypy. Measurements of the quality (Wiener entropy Tchernichovski et al. 2000) of these signals show a marked improvement over those calls recorded at dispersal age (M. C. Baker and D. E. Gammon, unpublished data). Once the over-wintering flocks have been formed, the gargle call repertoires in a local population can be examined.

Separating Ligand Bound from Ligand Free Complexes

We consider a situation where a ligand is nonstoichiometrically bound, whose mass is small compared to the target molecule. As an example, the mass of the tRNA (-70 kD) is just 3 of the total mass of the 70S ribosome (2.3 MD). With this assumption, the bias that is introduced by the ligand in the alignment and classification is negligible. This means that the orientation of the projection of a ligand-bound molecule is readily found by matching it to projections of a 3D reference. What follows is a description of a procedure that was developed and tested by Spahn and coworkers (C.M.T. Spahn, P.A. Penczek, and J. Frank, unpublished results).

From Anthesis to Senescence Setting the Stage

The development of the flower from anthesis to senescence often receives little attention. However, the internal changes that occur during this phase of development set the stage for flower senescence. It should be noted here that ephemeral flowers such as morning glory and daylily enter into senescence almost immediately following anthesis and therefore do not have the maintenance phase of flower development often observed in other species (Suttle and Kende, 1980 Bieleski and Reid, 1992 K. Smith and S. Verlinden, unpublished). S. Verlinden, unpublished). The maintenance phase is associated with a gradual decline in fresh weight, dry weight, carbon content and phosphorus content (Verlinden, 2003 K. Smith and S. Verlinden, unpublished).

TLR signaling pathways

In 800 genes in mouse macrophages within 3 h of LPS stimulation ( 10 of the genes examined) ( 82 and unpublished observations). Extending this observation, more recent microarray data suggests that LPS induces significantly more genes through the MyD88-independent than the MyD88-dependent pathway 83 . Another important feature of TLR4 regulation that has only recently come to light is that certain proteins, e.g., ST2, SIGIRR, Tollip, IRAK-M, MyD88s, SOCS, SHIP and others exert suppressive effects on TLR4 signaling (reviewed in 10, 81 Fig. 2). Thus, the regulation of TLR4 signaling represents a carefully orchestrated balance of positive and negative regulatory signals.

Potential explanations

At least three explanations could account for these data. The first is that these data represent simply a sampling artifact and that the 12 chickadees recorded in 2003 do not accurately represent the overall population. Because I sampled unbanded birds, it is also possible that the 2003 sample may contain mostly ASY birds with fully crystallized songs whereas the 2004 sample may have contained mostly SY birds with songs retaining some plasticity. Given that 35 to 45 of the Fort Collins populations were SY birds before and after the bottleneck (M. C. Baker, unpublished data), however, this possibility seems unlikely.

How postdispersal social environment may affect song development

Social pressure within a population might manifest itself to a developing bird in many ways, such as the number of its adult neighbors, its spatial proximity to these neighbors, and the number of countersinging interactions involving the young bird or overheard by the young bird. Any or all of these factors might play a role. In the 2004 Fort Collins and Islands North populations, chickadees experienced fewer adult neighbors, greater spatial isolation, and fewer countersinging interactions (unpublished data), and this may explain why the acoustic structure of their song types never became fully stereotyped. If countersinging interactions indeed have an effect on the level of song plasticity, it would be interesting to compare in an adult the amount of acoustic variation present during a countersinging interaction involving that bird versus the amount of variation present during its broadcast singing (e.g. during the dawn chorus). It would also be interesting to compare songs

The Ups In Huntingtons Disease

A recent study by Seo and co-workers analyzes, with fluorogenic substrates, the chymotrypsin and post-glutamyl activities of the proteasome in postmortem brain samples from HD patients and normal subjects (26). The chymotrypsin activity was decreased in the striatum of grade 0-1 and of grade 3-4 HD patients and also in the cerebellum of grade 0-1 HD patients. This activity did not change in cortex or substantia nigra of HD patients. The post-glutamyl activity was decreased in the striatum and cerebellum of grade 0-1 and of grade 3-4 HD patients, in the cortex of grade 0-1 HD patients and in the substantia nigra of grade 3-4 HD patients. For this analysis, individual proteasome activity values from post-mortem patients' brain samples were normalized using the atrophy index hat results from the macroscopic evaluation of ventricular size of the same HD patients' brains. Unfortunately, this study lacks measurement on total protea-some content in the brain samples, what is the key for the...

Injury by Reactive Oxygen Species

Such as nuclear condensation, nuclear fragmentation and plasma membrane vesiculation (J.S. Pober, unpublished observations). ECs that are sub-lethally injured by H2O2 are generally dysfunctional. However, chemical destruction of NO by H2O2 reduces the effectiveness of NO as a signalling molecule, resulting in endothelial dysfunction independent of actual cell injury.

Development of song in Parids

Investigators in the laboratory audio-tutored fledgling black-capped chickadees (Shackleton and Ratcliffe 1993 Kroodsma et al. 1995 K. A. Otter unpublished data) and fledgling Carolina chickadees (Kroodsma et al. 1995), and then monitored subsequent vocal development. For both species, high acoustic variation was found within juvenile song during early developmental stages, just as I found for juveniles in the field. This variation decreased over developmental time until the songs of males that were raised in the same group converged in acoustic structure (Shackleton and Ratcliffe 1993 Kroodsma et al. 1995). Interestingly, some of the acoustic features distinguishing the songs of adult black-capped chickadees in Fort Collins from typical fee-bee song (e.g. introductory syllables, novel amplitude breaks, and syllable recombination) also arose in the songs of black-capped chickadees reared in the laboratory with a limited social environment (Shackleton and...

Cf And Pancreatic Cancer

In a study on the expression of pancreatic tumor-associated mucin genes (MUC1 and MUC4) in the pancreatic tumor cell line derived from a CF patient (CFPAC1) and its CFTR corrected subline, CFPAC-PLJ-CFTR, we have reported that the overexpression of MUC4 is associated with CF phenotype.61 Also, no CFTR expression was detected in 12 (75 ) of the 16 pancreatic tumor cell lines tested, although pancreas is considered as a site of high CFTR expression. We have further confirmed the linkage between the CFTR defect and MUC4 overexpression, using short-interfering RNA (siRNA) mediated gene silencing of CFTR (Singh et al., unpublished data). MUC4 is a membrane bound mucin, which is aberrantly expressed in pancreatic tumor tissue and cancer cell lines, that has no detectable expression in the normal pancreas.62 Recently, we have also shown that the knockdown of MUC4 in a metastatic pancreatic tumor cell line, CD18 HPAF, results in reduced tumor growth both in vitro and in vivo and decreases the...

Nuclear Steroid Receptors

Cellular localization of the nPR within the teleost ovary has not been reported to date, although preliminary evidence indicates that the receptor is present in the follicle cells in Atlantic croaker (unpublished observation), in agreement with previous evidence suggesting that the nPR initiates ovulation in sciaenid fishes. 20p-S, the maturation-inducing steroid or substance (MIS) in the spotted sea trout, was shown to induce ovulation in vitro in a dose-dependent manner (Pinter and Thomas, 1999). In addition, 20P-S-stimulated ovulation was inhibited by the addition of blockers of RNA (actinomycin D) and protein (cycloheximide) synthesis, indicating that progestin induction of ovulation is via a genomic mechanism. The third piece of evidence indicating that the nPR mediates ovulation in this species is that 17,20pDHP is a more potent inducer of ovulation than 20P-S. Binding studies on the sea trout nPR shows that 20P-S has a relative binding affinity for the nuclear receptor only 65...

Involvement of Protein Chaperones and the UPS in Turnover of Mutant SOD1 Proteins

Based on these measures of disease-progression, proteasomal enzyme activities were measured at three time points in homogenates of lumbar spinal cord, thoracic spinal cord and liver from SODWT and SODG93A mice and their nontransgenic littermates (59) day 45, when microscopic changes are present, but there is no significant alteration in cell type (either motor neuron death or glial activation) day 75, when motor neuron loss and glial activation are beginning, and at symptomatic age (around day 120). Assays of individual 20S 26S proteasome hydrolytic activities were based on generation of the fluorescent species, amino methyl coumarin (AMC) following addition of peptide substrates to samples of tissue homogenates (Suc-LLVY-AMC, Boc-LRR-AMC or Z-LLE-AMC as substrates for chymotrypsin-, trypsin-, and caspase-like activities, respectively). Total (normalized to actin) and specific activities (normalized to level of 20S proteasome core a-subunits) were determined. Both total and specific...

Minerals And Calcitropic Hormones

Serum phosphate levels are higher in the fetus than the mother in mammals (1), which may indicate that phosphate is actively transported across the placenta however, the regulators of phosphate transport are unknown. Both PTHrP and PTH are needed because the serum phosphate level is increased in Pthrp-null and Hoxa3-null (aparathyroid) fetuses (unpublished data ref. 4). 1,25-D has been suggested to play a role in placental phosphate transfer (9) however, in Vdr-null fetuses, the serum phosphate levels and skeletal mineral content are normal (7). Fetal magnesium metabolism has not been extensively studied, but the fetal magnesium level is at most slightly higher than the maternal magnesium concentration in most studies (1). Although absence of PTHrP in the Pthrp-null fetuses does not affect serum magnesium (unpublished data), the absence of parathyroids and PTH (Hoxa3 null) results in hypomagnesemia (4) and reduced skeletal magnesium content (5). The absence of calcitonin (Ct-null...

Placental Calcium Transport

Maternal hormones might influence fetal-placental calcium transport by raising or lowering the ambient maternal calcium level and by direct effects on the placenta. However, at least in animal models, a normal rate of maternal-to-fetal calcium transfer can usually be maintained despite the presence of maternal hypocalcemia or hormone deficiencies. Whether the same is true for human pregnancies is less certain. As examples, maternal hypocalcemia caused by parathyroidectomy or dietary calcium restriction did not affect the rate of fetal-placental calcium transfer as directly assessed in placental perfusion experiments in sheep (19). Similarly, in Vdr-null fetal mice, placental calcium transfer was normal even though the Vdr-null mothers were severely hypocalcemic (unpublished data ref. 7). However, a normal rate of maternal-fetal calcium transfer does not necessarily imply that the fetus is unaffected by the maternal hypocalcemia. The fetal-placental unit must be working harder to...

Comparison Of The Toxicity Of Mutant Sod1 And Polyqexpanded Ar

Because mutations in SOD1 and AR both cause MND, identifying common mechanisms underlying the vulnerability of motor neurons to toxicity could have therapeutic implications. Because HSPs reduce aggregation of both types of mutant protein (45, 46, 110, 115, 116), the high threshold for activation of the heat shock response may be a contributing factor (12). In cultured motor neurons, toxicity of both mutant SOD1 and polyQ-expanded AR are reduced by treatment with AMPA receptor antagonists and by overexpression of the cytoso-lic calcium-binding protein calbindin (11) (Tradewell and Durham, unpublished data), indicating even normally non-toxic levels of glutamate receptor activation sensitize motor neurons in a calcium-dependent manner.

Implications For Therapy Of

If depletion of protein chaperones and inhibition of proteasomes are significant in the pathogenesis of protein folding diseases, would upregulating these pathways be a reasonable therapeutic strategy On the protein chaperoning side, our studies indicate that coordinate upregulation of multiple HSPs, particularly those that work coordinately such as HSP70 and HSP40, is more likely to be beneficial than a single HSP (Batulan et al., unpublished results). Many drugs known to magnify the heat shock response mediated through the transcription factor HSF1 are not particularly effective in cultured motor neurons or are too toxic (12). However, recent studies show potential for hydroxy-lamine derivatives (48) and celastrol, a quinone methide triterpene and an active component from Chinese herbal medicine (128) as regulators of the heat shock response in an as needed manner. Other components of the UPS (e.g., ubiquitin E3 ligases) that can sequester mutant proteins until they can be degraded...

Relevance of TIR8 inhibitory effects in inflammatory conditions in vivo

Uation of intestinal inflammation by bacterial products from the lumen, activation of DC and of a T cell mediated colitis 35 . Recognition of microbial moieties of the enteric flora and production of inflammatory cytokines, in particular IL-1a and IL-18, play a key role in intestinal inflammation both in experimental systems and in humans 36-39 . TIR8 deficient mice showed increased severity of colitis induced by DSS, in terms of local tissue damage and mortality ( 22 and X. Li, unpublished). The observation of increased severity of colitis in TIR8 deficient mice is consistent with a non-redundant regulatory role of this molecule in the gastrointestinal mucosa. It is conceivable that increased production of inflammatory cytokines in response to tissue damage and exposure to microbial molecules by DC in the lamina propria and possibly by epithelial cells is responsible for a more severe colon inflammation in Tir8- - mice.

Hydroxytryptamine2 Receptors

Turner and Raymond reported that the 5-HT2A receptor possesses two CaM-binding domains in the i2 loop and proximal juxtamembrane region of the car-boxyl terminus (199). Both sites possess putative a-helical structures with positively charged amino acids located on the face of the helix opposite from hydrophobic residues, consistent with CaM-binding motifs. The affinities of CaM for those synthetic peptides derived from those regions of the receptor are approx 65 nM and approx 170 nM, respectively, suggesting that the interaction between CaM and the 5-HT2A receptor is relevant. CaM coimmunoprecipitates with the receptor and binds avidly to fusion proteins containing each of the putative CaM-binding regions of the 5-HT2A receptor. CaM inhibits G protein coupling by the 5-HT2A receptor and decreases phosphorylation of the 5-HT2A receptor carboxyl-terminal peptide. Those findings suggest that CaM might dampen G protein coupling primarily through binding to the i2 loop and attenuate...

Molecular Dynamics Simulations

We have confirmed by molecular dynamics (MD) simulations that large conformational changes of the observed type can indeed be triggered by protonation of His163 and deprotonation of His172.10 In three different 10-ns simulations at pH 6.0, 7.6, and 8.0, we found the same type of rearrangements as seen in the crystals. This is reassuring, because it should not be forgotten that unless the crystals diffract to better than 1.0 A resolution, X-ray diffraction cannot normally determine hydrogen positions because of their low scattering power. The force fields used in MD simulations, on the other hand, fully take into account the hydrogens bound to non-carbon atoms. Thus, because these simulations yield the same conformational rearrangements as those seen by X-ray diffraction, the interpretation of the crystallographic results is likely to be correct. Also, our preliminary NMR data with 15N-labeled SARS-CoV Mpro (J. George et al, unpublished) appear to support these conclusions.

Dna Replication Proteins

In addition, a homologue to the middle subunit (CpRPA2) was recently identified from the CpGP. Despite the low homology among all RPA2 subunits, we were able to confirm that CpRPA2 protein was indeed an ssDNA-binding protein and could be regulated by phosphorylation. The phosphorylation of CpRPA2 completely abolished the ssDNA-binding property. Similar to other DNA replication proteins, the expression of CpRPA2 gene is regulated by the cell cycle and its protein is distributed more intensively in the C. parvum oocysts, meronts and gamonts than other life cycle stages (Millership and Zhu, unpublished data).

Sex hormones and the hippocampus

The problem with all of this plasticity in CA1 pyramidal neurons is that, as noted earlier, the only detectable intracellular estrogen receptors are found in interneu-rons, not in the CA1 pyramidal neurons themselves. The possible exception is the presence of the beta ER in CA1 and CA2 neurons, discussed previously. There are also no detectable estrogen-inducible intracellular progestin receptors in hippocampus, based upon immunocyto-chemistry (N. G. Weiland and S. Alves, unpublished) however, there is a binding study showing low levels of such receptors in hippocampal tissue (Parsons et al., 1982). Nevertheless, recent studies with an estrogen antagonist argue strongly for a role of alpha estrogen receptors in interneurons in synapse induction. The estrogen antagonist CI-628 blocks estrogen induction of spines on CA1 neurons and does not produce any agonist effect (McEwen, Tanapat, and Weiland, 1999). This situation is reminiscent of blockade of progestin receptor induction in...

Adrenal steroids and the hippocampus

More recent studies have begun to dissect the underlying mechanism and the role of Type I and Type II receptors. Within the dentate gyrus, LTP can be modulated biphasically by adrenal steroids acting via Type I and Type II receptors LTP was found to be enhanced within an hour by adrenal steroids acting via Type I receptors, whereas Type II receptor stimulation was found to rapidly suppress LTP (Pavlides, Watanabe, et al., 1995). The enhancement by Type I receptor stimulation lasted for more than 24 hours in awake freely moving animals, whereas LTP disappeared in a few hours in the dentate gyrus of adrenalectomized rats without steroid replacement (Pavlides et al., 1994). The CA1 field responded to corticosteroids in a biphasic manner similar to that of the dentate gyrus (Pavlides et al., 1996) and the same was true of the commissural associational input to CA3 but not for the mossy fiber input (C. Pavlides and B. McEwen, unpublished). These biphasic effects may help to explain the...

Iliac Vein Stent Placement in PTS Reflux

Iliac vein stent placement recently has emerged as effective therapy in PTS.39 A surprising initial finding was relief of symptoms including healing of stasis ulceration (55 cumulative near term) with stent placement alone even when the associated reflux remained uncorrected.40 These unexpected results appear to have been sustained over a longer follow up of > 4 years as well (unpublished data). This has important implications for the management of PTS as the majority have combined obstruction reflux.1013,39 Most patients should benefit from stent placement alone,

Grapevine Fanleaf Virus and Tomato Ringspot Virus

In view of this hypothesis, stretches of hydrophobic residues were identified within the 1BHel from both GFLV and ToRSV (Rott et al. 1995 Ritzenthaler, unpublished results). Thus, the 1BHel from ToRSV, also named nucleoside triphosphate-binding (NTB) protein, contains a hydrophobic region at its C terminus consisting of two adjacent stretches of hydrophobic amino acids separated by a few amino acids. Sanfa on and colleagues recently showed that 1BHelVPg associates with canine microsomal membranes in the absence of other viral proteins in vitro and with ER membranes in planta (Wang et al. 2004). Analysis performed on truncated proteins fused to GFP confirmed the presence of a functional transmembrane domain within the 60 amino acids at the C terminus of protein 1B and also revealed the presence of a putative amphipathic helix within the N-terminal 80 amino acids of 1B (Zhang et al. 2005). In agreement with the replication of ToRSV on ER-derived membranes (Han and...

Endocytic Function of PSMA

Psma Internalization

Internalization of membrane proteins from plasma membrane and sort these proteins to endosomes lysosomes and, finally, to the compartment for peptide loading (35-37). These sorting signals are tyrosine-based NPXY and YXXO motifs (37) and dileucine motif (36). Tyrosine motifs are identified in a variety of receptor molecules like the transferrin receptor, low-density lipoprotein receptor, and asialoglycoprotein receptor (38). Dileucine (or leucine-isoleucine sequence) motif (Fig. 3A and Color Plate 12 following p. 302) is important for internalization and lysosomal targeting was found in the y-5 chain of the T-cell receptor, CD4, IFN-y (39-41). Tyrosine-based motifs interact with adaptor complexes API, AP2, and AP3 (42,43) and dileucine-based motifs bind to the P-subunits of API and AP2 -chains of API and AP2 has also been reported to bind to these signals. Apart from this, leucine-based signals of lysosomal protein LIMPII and melanosomal membrane protein tyrosinase have been shown to...

Novel pathways regulated during e2 senescence

As described earlier, cervical carcinogenesis depends on the continuous expression of the viral E6 and E7 oncogenes in the affected individual. Transcription of E6 E7 is negatively regulated by E2 and this repression is both necessary and sufficient for a reversal of transformation via senescence (114,138). This argues, in turn, that important senescence mediators must be inhibited by the viral oncoproteins at the onset and during maintenance ofHPV-positive carcinogenesis. Regulation ofspecific genes during E2 senescence might, thus, help elucidate how HPV oncoproteins target important senescence regulators and might uncover basic molecular relationships between HPV carcinogenesis and senescence. Immortalization of human primary foreskin keratinocytes by E6 and E7 reflects certain aspects of carcinogenesis in vivo and, indeed, a number of senescence-associated genes as identified earlier have already turned out to be novel transcriptional targets ofHPV oncogenes in this model system...

Bimodal exploration and haptic abilities

Examining under visual control is used at the same time as oral exploration. It involves different exploratory activities, such as skilled manipulation, applying pressure, transferring from one hand to the other or rotating the object. However, the two modes of functioning of the haptic system evolve differently when they are under visual control. Oral exploration, which performs well at the age of five months, peaks at seven months, then declines until eleven months. On the other hand, the tendency to examine objects manually and visually progresses and supplants oral exploration. After seven months, the baby rediscovers its capacity for subtly manipulating objects and extracting information like shape, texture and substance. According to the authors, this late substitution is explained by the fact that between the ages of five and seven months the visual system is busy checking newly emerging skills like reaching for and grasping objects. The hands can therefore no longer fulfill...

Regulation of PSMA Expression by PSMA Promoter Enhancer

Psma Prostate Cancer

Ca2+ ions positively regulate PSMA expression. Ca2+ influx probably takes place through a CaT-like calcium channel, which is strikingly correlated with the malignancy of prostate cancer as well as PSMA expression (61). How could that start The possible explanation could involve glutamate receptors. We have observed metabotropic glutamate receptors by gene-array analysis of LNCaP cells (Heston et al., unpublished observation). In prostate cancer cells, such receptors could get activated constitutively by free glutamates (an agonist to such a receptor) released as a byproduct of the folate hydrolase NAALADase action of PSMA expressed on the cell surface (PSMA level is upregulated many-fold in prostate cancers) and can modulate the function of potassium and calcium channels, which might cause change in resting membrane potential. Such a change in membrane potential could cause oxidative damage to the cells, release of Cl- ions (62), and continuous influx of Ca2+ ions through calcium...

The Hypothalamuspituitaryadrenal Hpa Axis

Difference Between Raas And Hpa Axis

Adrenalectomy did not alter orexin immunoreactivity in the hypothalamus (Taheri, unpublished data). Stricker-Krongrad and Beck (51) have, however, reported that with adrenalectomy there is a 50 decrease in lateral hypothalamic prepro-orexin mRNA detected by in situ hybridization. This returned to normal with dexamethasone administration. The reason for the discrepancy between peptide and mRNA levels is unclear.

Neurophysiological Studies Of Tactile Attention

FIGURE 8.5 Effects of attention on orientation tuning curves. Response of a neuron in SII to oriented bars while performing a tactile and visual task. During the tactile task, the bar indented the skin at one of eight randomly chosen orientations (S1) for 500 msec. After a 1 second delay, the bar indented the skin for an additional 500 msec at either the same orientation or at the orthogonal orientation (S2). The animal's task was to pull a switch with its foot if the orientations of the two bars were the same or to push the switch if the orientations were different. Raster plots on the left show the responses sorted according to bar orientation and task. The curves to the right show the mean impulse rates as a function of orientation. (Hsiao, unpublished data.) FIGURE 8.5 Effects of attention on orientation tuning curves. Response of a neuron in SII to oriented bars while performing a tactile and visual task. During the tactile task, the bar indented the skin at one of eight randomly...

Type Of Pancreatitis Initial Damage In The Pancreas And Fibrosis Pattern

It has recently been shown that the genetic alterations in hereditary chronic pancreatitis involve the cationic trypsinogen gene (PRSS1) or a serine protease inhibitor gene (SPINK1).22-24 Mutations in these two genes seem to trigger the autoactivation of trypsinogen in the pancreas, which probably results in the early inappropriate conversion of pancreatic zymogens to active enzymes in pancreatic ducts with subsequent autodigestive necrosis and inflammation. The site of the autodigestive necrosis may be either the acinar cell or, more likely, the duct wall. In our series of pancreatic resection specimens from six patients with hereditary chronic pancreatitis (G.K., unpublished observations), we found advanced chronic pancreatitis with severe periductal fibrosis and massively dilated ducts containing protein plugs and calculi staining for pancreatic stone protein (PSP). In one case, there was ductal necrosis in a number of medium-sized interlobular ducts that destroyed the duct...

Effectiveness Of Endovenous Laser Treatment

Durable occlusion of the treated saphenous vein segment is certainly a central goal of endovenous laser treatment. However, recanalization of the Great Saphenous vein can be observed and frequently it seems to be associated with low energy delivery during treatment.15 Figure 29.3 displays a so-far unpublished plot of otherwise published original data about three months follow-up of continuous laser treatment of Great Saphenous veins.15 It clearly displays that recana-lization is associated with low LEED values, and furthermore that veins with greater diameter require the delivery of more energy to stay occluded during the first three months after treatment. The line drawn in Figure 29.3 is the result of linear regression analysis of open boxes showing a slope of about 10 joule per cm GSV diameter. This indicates that

TLR ligands during allergic sensitization

Part of the apparent contradiction among these different epidemiologic findings might result from opposing actions of TLR ligands to the sensitization of allergens and responses to them. In principle, these two aspects of TLR function can be addressed experimentally in animal models of asthma. However, even when the effect of TLR ligands is restricted to sensitization observed results have varied, depending on experimental conditions. In the most common model of ovalbumin-induced allergic inflammation, mice are sensitized by an intraperitoneal (i.p) injection of ovalbumin complexed with the adjuvant aluminum hydroxide (alum), which promotes TH2 responses. In rats and Balb c mice, exposure to LPS at the time of ovalbumin sensitization diminishes subsequent allergic responses in the lung following antigen challenge 76, 77 . The strain of mice used might be relevant to this experiment because we have not observed an effect of i.p. injection of LPS at the time of sensitization in a...

Envelope Switching Flexibility of Rhabdovirus Envelopes

The propensity of rhabdoviruses to accept changes in the composition of the envelope is of great potential for manifold vector applications. Rhabdoviruses carrying novel proteins in their envelopes may have a role to play as particulate vaccines, and as targetable gene delivery vectors (Table 2). Early experiments with ts mutants of VSV (Schnitzer et al. 1979) initially challenged the importance of the rhabdovirus G protein for virus particle formation. The generation of recombinant RV lacking the G gene then confirmed that rhabdovirus particle formation does not require G, though in the presence of G, budding efficiency is increased approximately 30-fold (Mebatsion et al. 1996a). The major driving force for rhabdovirus budding is thus the M protein, as was also confirmed by analysis of a rabies virus M gene deletion mutant (Mebatsion et al. 1999 Finke and Conzelmann 2003). Moreover, it appears that the M proteins of rhabdoviruses alone, when expressed in the absence of any other...

Energy consumption for development

Fig. 2 Specific heat production rates of honeybee drones (Apis mellifera) during development. Bars indicate standard deviations. Abbreviations L2, L3, L7 2nd, 3rd, 7th day of larval development V1, V2 day 1 and 2 of prepupal stage H moulting from larva to pupa H moult from pupa to adultus P1-P8 Day 1-8 of pupal development RA resting adult between moult to adult and hatch from the capped cell S hatching adult A young adult (age < 24h). Unpublished data from the authors Fig. 2 Specific heat production rates of honeybee drones (Apis mellifera) during development. Bars indicate standard deviations. Abbreviations L2, L3, L7 2nd, 3rd, 7th day of larval development V1, V2 day 1 and 2 of prepupal stage H moulting from larva to pupa H moult from pupa to adultus P1-P8 Day 1-8 of pupal development RA resting adult between moult to adult and hatch from the capped cell S hatching adult A young adult (age < 24h). Unpublished data from the authors Fig. 4 Heat production rate of a honeybee...

Other Genital Hygiene Practices

Wet wipes are gaining popularity in North America and Western Europe. In the California study cited previously, usage rose with age from 26 among women younger than age 41 to 40 among women older than age 48 (52). Such products are often used more than once a day. Baby wipes, premoistened toilet wipes, and feminine wipes are all common choices (Farage and Bramante, unpublished data, 2004). In the late 1980s, reports appeared of allergic contact dermatitis to preservatives in some European wipes (138). The preservative in question (methylchloroisothiazolinone) is now highly regulated. Moreover, quantitative sensitization risk assessments have progressed over the last 20 years, such that it is now possible to safely formulate consumer products containing such preservatives at levels so low that they pose no significant risk of inducing contact sensitization. Pubic hair removal is performed in some countries. In response to a survey of 635 Turkish women, 98 reported pubic hair removal on...

Clinical trial results with avastin 21 Efficacy

Response rate, duration of response, and survival without significant toxicity. Avastin added to 5-FU LV appeared to have significant activity as first-line therapy for metastatic colorectal cancer. These results, along with a completed phase II study of combining Avastin with 5-FU LV (8) and a second phase II trial of Avastin in combination with 5-FU LV for patients that are not optimal candidates for irinotecan (data unpublished), demonstrate that combining Avastin with either 5-FU- or irinotecan-containing chemotherapy regimens provides substantial survival benefit for first line treatment of metastatic colorectal cancer.

Development of corticocortical pathways

During phylogeny, there is a progressive shifting of visual processing to more central structures of the brain. One consequence is that, in the mammal, more than 90 of the synapses in the brain occur in the cerebral cortex. Quantitative investigation of the cortex shows that 95 or more of synapses in the cortex are of cortical origin (Kennedy and Falchier, unpublished). One consequence of these figures is that developmental processes of the corticocortical pathways could have a major impact on the development of visual function.

Endocrine And Paracrine Regulation

Caspase-3, have been shown to be present in the ovary of mammals and birds (reviewed in Andreu-Vieyra and Habibi, 2000 Johnson and Bridgham, 2000). Although caspase tissue distribution in teleost gonads remains unknown, we have recently detected caspase-3 activity in the goldfish testes and ovary (Andreu-Vieyra et al., 2005 Andreu-Vieyra and Habibi, unpublished data). More recently, studies have focused on the regulation of apoptosis in the ovary and several factors have been shown to provide protection against gonadal cell apoptosis under different conditions (Andreu-Vieyra and Habibi, 2000). For example, GtHs and E2 have been shown to prevent apoptosis in the mammalian ovary under a variety of conditions (Billig et al., 1993 Chun et al., 1996 Kaipia and Hsueh, 1997 McGee et al., 1997 Andreu-Vieyra and Habibi, 2000 Markstrom et al., 2002 Wood and Van der Kraak, 2002). Similarly, in vitro treatment of rainbow trout follicles with epidermal growth factor, E2, and gonadotropins has been...

Speech Perception Indices

The first credible effort to evaluate the effectiveness of a communication system based on physical measurements was that of H. Fletcher (1921). In an unpublished Western Electric Laboratory report, Fletcher pointed out that a suitable measure (index) of circuit quality must have the property of additivity. By this he meant that if a particular frequency range of speech heard alone has a quality value of Q1 and if a second frequency range has a value Q2, then the value of both ranges heard at the same time should equal Q1 + Q2. It was clear that articulation test scores did not possess this In a parallel development in England, John Collard, working for International Telephone and Telegraph, published a detailed description of an index with properties similar to those of the articulation index in a series of papers beginning in 1930. He called his index band articulation (or frequently just ''the new unit''). Speech test scores were called sound articulation. By 1939 Col-lard had...

Other nutritional changes 1471 Antinutrients

The health benefits of phenolic acids and flavonoids are being actively studied today. Potato peels free phenolics, primarily chlorogenic acid, were reduced by extrusion (unpublished data, Camire and Dougherty), with improved retention at higher barrel temperature and feed moisture. Blueberry and grape anthocyanins were significantly reduced by extrusion and by ascorbic acid in sweetened corn breakfast cereals (Chaovanalikit, 1999).

Mapping Maternal Effect Mutations

Fine mapping a maternal-eVect mutation can be performed much more eYciently than the initial mapping. Narrowing down the location of a mutation through fine mapping is necessary to identify the molecular nature of the mutated gene through either candidate gene or positional cloning approaches. Once a maternal-eVect mutation is mapped, homozygous male and heterozygous female carriers can then be identified by polymorphic markers flanking the mutation. Such F0 fish are intercrossed to map finely the position of the mutation in the F1 progeny. In a cross of a heterozygous female to a homozygous male, mapping is performed with recombinants generated only through meiosis in the F0 female, because the male is homozygous for the mutation. Thus, each F1 fish represents a single meiosis rather than two meioses for progeny from heterozygote intercrosses. However, the loss of recombination events from the homozygous male is oVset by meiotic recombination being suppressed in males compared to...

Attenuation of Rhabdoviruses

A pre-condition for reasonable use of such gene deletion vectors is a powerful complementation system. Since both rhabdovirus M and G proteins are cytopathic, the generation of packaging cell lines is a problem. However, the use of tight conditional expression, such as the Tet-on off system, has allowed the establishment of packaging cells in which, for example, expression of RV M protein, or both M and G, can be induced (Finke et al. 2003). In these systems, production of M and or G-deleted RV titers of up to 107 infectious virus particles per milliliter supernatant (Finke, unpublished results).

Clinical manifestations

The natural history of DM is unknown, as most patients nowadays are treated with steroids. The mortality rates reported 20-30 years ago are outdated. Clinical experience indicates that DM responds to therapy more readily than polymyositis. In children, DM may at times be a monophasic disease with infrequent flares once the disease is under control. We have seen, however, late progression in some patients, including the development of inclusion-body myositis in two patients (Dalakas, unpublished observations). Patients with interstitial lung disease may have a high mortality rate, requiring aggressive treatment with cyclophosphamide or Tacrolimus. Overall, there are still a number of patients who do not respond

Pancreatic Pathways Molecular Epidemiology And The Study Of Geneenvironment Interactions

Null (homozygous deletion) genotype and pancreatic cancer, nor did we find any evidence for interactions between cigarette smoking and the GSTM1-nuVl genotype.6 We did find evidence for interactions between cigarette smoking and GSTT1-nuVl,6 and between cigarette smoking and GSTP1-105Val (Duell and Holly, unpublished data) in relation to pancreatic cancer risk estimates. Interestingly, GSTT1 smoking interactions were stronger among women, whereas GSTP1 smoking interactions were stronger among men. These differences could reflect biological differences in expression and activity, or differences in smoking behavior or other exposures in nonsmokers (such as occupational exposures). CCK is a gastrointestinal hormone known to influence normal and malignant pancreatic growth and function (see also Chapter 6 and Chapter 14). A number of polymorphisms in genes for CCK and CCK receptors have been identified. Because of the importance of CCK in pancreatic growth and function, we hypothesized...

Endocrine Disruption Of Nongenomic Steroid Actions

Interestingly, several xenoestrogens have been shown to interfere with progestin stimulation of sperm motility in this species by the same receptor-mediated mechanism (Thomas et al., 1998 Thomas and Doughty, 2004). The identification of the novel mPR involved in oocyte maturation enables the interactions of xenoestrogens with membrane steroid receptors to be investigated for the first time at the molecular level. Xenoestrogens also have the potential to interfere with the actions of estrogens through binding to the membrane estrogen receptor. The ortho, para derivative of DDT (o',p'-DDE), binds to membrane estrogen receptor recently identified in human breast cancer cells, GPR30, and has estrogenic activity in cells expressing the receptor (Thomas, 2006). Recently GPR30 has been detected in croaker ovaries (unpublished observation). In croaker testes the decrease in androgen production by estrogens was mimicked by Kepone, nonylphenol, and 2,2', 5'-PCB-4-OH....

Insights From Animal Ace2 And Animalderived Viral Isolates

The ability of the ACE2 proteins of mice, rats, and palm civets to support SARS-CoV infection has been compared with that of human ACE2.22, 51 Compared with cells expressing human receptor, SARS-CoV infection was less efficient in cells expressing murine ACE2. Infection was nearly absent in those expressing rat ACE2. Consistent with a role for palm civets in transmitting virus, palm civet ACE2 supported SARS-CoV infection as efficiently as human ACE2. These results correlated with affinity of each of these receptors for the S protein and its RBD.22, 51 Chimeras between human and rat ACE2 receptors were used to identify the S-protein binding site on ACE2.22 Mutation of four rat ACE2 residues (82-84 and 353) to their human equivalents converted rat ACE2 into an efficient SARS-CoV receptor. Residues 82-84 comprise a glycosylation site on the rat receptor that is not present on mouse, palm civet, or human receptor. Residue 353 is a histidine in mouse and rat receptors, and a lysine in...

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