Screening of phenylalanine ammonia lyase in plant tissues,and retention of activity during dehydration

BACKGROUND: Oral therapy with phenylalanine ammonia lyase (PAL), naturally encapsulated in plant cells, may provide a potential alternative treatment for hyperphenylalaninemic patients, including those with phenylketonuria. Therefore different sources of plant tissue were investigated for PAL activity. RESULTS: Enzyme activity was highest in grain seedlings, with maximal enzyme activity in 7‐day‐old red spring wheat (Triticum aestivum L.) seedlings. The PAL activities of leaves and roots/endosperm of wheat seedlings were 11.90 ± 2.64 and 6.48 ± 1.59 µmol h^?1 g^?1 dry weight respectively. Three PAL‐related polypeptides with molecular weights of 74, 83 and 103 kDa were identified in wheat seedling leaf tissues, while only the 74 kDa polypeptide was detected in root/endosperm tissues. Dehydration was investigated as a method of concentrating PAL in wheat seedlings. Freeze‐drying was found to retain the most PAL activity (>90% recovery on a dry weight basis) compared with air drying and vacuum microwave drying for both leaf and root/endosperm samples. CONCLUSION: This study has led to a better understanding of PAL activity and stability in plant tissues and provides the basis for developing a natural plant preparation as a dietary supplement for the treatment of hyperphenylalaninemia. Copyright © 2007 Society of Chemical Industry     

Preischaemic as well as postischaemic application of a calcium antagonist affords cardioprotection in the isolated guinea pig heart

OBJECTIVE: The aim was to answer the following questions: (1) Does treatment with calcium antagonists have to be begun before ischaemia or is postischaemic application also protective? (2) When applied before ischaemia, do calcium antagonists have to depress preischaemic cardiac function in order to elicit protection? (3) Is cardioprotection a matter of improved reflow or do the agents influence the degree of oxidative injury during reperfusion? METHODS: Isolated working guinea pig hearts underwent ischaemia (15 min) and reperfusion (15 min). The calcium antagonist gallopamil was given either before (0.1 nM and 1 nM) or after ischaemia (0.1 nM) during early reperfusion (first 5 min). Recovery was defined as postischaemic compared to preischaemic external heart work, expressed in percent. Oxidative stress was assessed by the release of glutathione (GSH). Lactate release served as a measure of the ischaemic challenge. The ability of gallopamil to scavenge oxygen radicals directly was investigated in an in vitro chemiluminescence assay. RESULTS: Pump function of control hearts recovered to only 28% after reperfusion. Pretreatment with 0.1 and 1 nM gallopamil improved recovery to the same extent (48.7% and 43.4%, respectively); however, postischaemic application of 0.1 nM gallopamil afforded equal protection (45.4% recovery). Only the higher concentration of 1 nM gallopamil depressed preischaemic external heart work (by 11%). During earliest reperfusion (1-5 min), release of GSH only tended to be lower in treated hearts. During the subsequent minutes of reperfusion (5-15 min), release of GSH was significantly less in hearts postischaemically treated with 0.1 nM gallopamil (40 pmol.min-1 v 940 pmol.min-1 for controls). In contrast, ischaemia-induced lactate release did not differ between the groups. Gallopamil did not scavenge reactive oxygen species in vitro. CONCLUSIONS: Short term postischaemic application of the calcium antagonist gallopamil is almost as effective at restoring pump function as preischaemic application which, in turn, does not have to depress preischaemic cardiac function in order to elicit protection. A reduction of oxidative stress during reperfusion seems to contribute to the beneficial effects of postischaemic application of gallopamil, but a direct oxygen radical scavenging activity of gallopamil is not involved.     

Relations Among Work-Home Culture, the Utilization of Work-Home Arrangements, and Work-Home Interference.

In the present study, we examined the associations among work-home culture (WHC), the utilization of work-home arrangements (WHAs), and work-home interference (WHI) among 638 workers from a Dutch financial consultancy firm. We (a) developed a typology of WHC, (b) examined whether the utilization of 6 WHAs differed for various types of WHC, (c) determined whether various types of WHC and the utilization of WHAs were related to WHI, and (d) studied these associations for subgroups of workers. Results showed that WHCs can be characterized by 2 dimensions, i.e., support and hindrance. More supportive and less hindering WHCs were not associated with a higher utilization of WHAs, but did covary with lower levels of WHI. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Psychometric properties of the Questionnaire on Stress in patients with Diabetes—Revised (QSD-R).

This study reports the psychometric properties of a 45-item diabetes-specific questionnaire, the Questionnaire on Stress in Patients with Diabetes—Revised (QSD-R), a modified and shortened version of the QSD (G. Duran et al; see record 1996-18805-001). The QSD-R was filled out by 1,930 individuals with insulin-dependent diabetes mellitus and noninsulin-dependent diabetes mellitus. Eight consistent scales were identified (values of Cronbach's α: .69–.81). The test-retest reliability for the total score after a 5-week interval was rtt?=?.63. The results provide evidence for the reliability and validity of this instrument. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

High dynamic range image rendering with a Retinex-based adaptive filter.

We propose a new method to render high dynamic range images that models global and local adaptation of the human visual system. Our method is based on the center-surround Retinex model. The novelties of our method is first to use an adaptive filter, whose shape follows the image high-contrast edges, thus reducing halo artifacts common to other methods. Second, only the luminance channel is processed, which is defined by the first component of a principal component analysis. Principal component analysis provides orthogonality between channels and thus reduces the chromatic changes caused by the modification of luminance. We show that our method efficiently renders high dynamic range images and we compare our results with the current state of the art.     

Overgeneral memory extends to pictorial retrieval cues and correlates with cognitive features in posttraumatic stress disorder.

Individuals with posttraumatic stress disorder (PTSD) show overgeneral memory (OGM) when retrieving autobiographical memories to word cues. We investigated whether OGM extends to picture cues and whether it is related to PTSD symptoms and cognitions. Trauma survivors with (n = 29) and without (n = 26) PTSD completed the standard Autobiographical Memory Test (AMT) and a novel picture version. Compared to the no-PTSD group, the PTSD group showed OGM in both test versions. Pictures facilitated specific memory retrieval, but this effect was no longer significant when verbal intelligence or depressive symptoms were controlled. OGM correlated with PTSD symptoms and perceived self-change; with intrusive memories, their perceived "nowness," responses to intrusions (thought suppression, rumination, dissociation), and negative interpretations of symptoms. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Making the Bend: DNA Tertiary Structure and Protein-DNA Interactions

DNA structure functions as an overlapping code to the DNA sequence. Rapid progress in understanding the role of DNA structure in gene regulation, DNA damage recognition and genome stability has been made. The three dimensional structure of both proteins and DNA plays a crucial role for their specific interaction, and proteins can recognise the chemical signature of DNA sequence (“base readout”) as well as the intrinsic DNA structure (“shape recognition”). These recognition mechanisms do not exist in isolation but, depending on the individual interaction partners, are combined to various extents. Driving force for the interaction between protein and DNA remain the unique thermodynamics of each individual DNA-protein pair. In this review we focus on the structures and conformations adopted by DNA, both influenced by and influencing the specific interaction with the corresponding protein binding partner, as well as their underlying thermodynamics.     

The G-Protein-Coupled Estrogen Receptor (GPER/GPR30) in Ovarian Granulosa Cell Tumors

Ovarian granulosa cell tumors (GCTs) are thought to arise from cells of the ovarian follicle and comprise a rare entity of ovarian masses. We recently identified the G-protein-coupled estrogen receptor (GPER/GPR30) to be present in granulosa cells, to be regulated by gonadotropins in epithelial ovarian cancer and to be differentially expressed throughout folliculogenesis. Thus, supposing a possible role of GPER in GCTs, this study aimed to analyze GPER in GCTs. GPER immunoreactivity in GCTs (n = 26; n (primary diagnosis) = 15, n (recurrence) = 11) was studied and correlated with the main clinicopathological variables. Positive GPER staining was identified in 53.8% (14/26) of GCTs and there was no significant relation of GPER with tumor size or lymph node status. Those cases presenting with strong GPER intensity at primary diagnosis showed a significant reduced overall survival (p = 0.002). Due to the fact that GPER is regulated by estrogens, as well as gonadotropins, GPER may also be affected by endocrine therapies applied to GCT patients. Moreover, with our data supposing GPER to be associated with GCT prognosis, GPER might be considered as a possible confounder when assessing the efficacy of hormone-based therapeutic approaches in GCTs.     

Cellular Internalisation of an Inositol Phosphate Visualised by Using Fluorescent InsP5

When applied extracellularly, myo‐inositol hexakisphosphate (InsP₆) and myo‐inositol pentakisphosphate (InsP₅) can inhibit the growth and proliferation of tumour cells. There is debate about whether these effects result from interactions of InsP₆ and InsP₅ with intracellular or extracellular targets. We synthesised FAM‐InsP₅, a fluorescent conjugate of InsP₅ that allows direct visualisation of its interaction with cells. FAM‐InsP₅ was internalised by H1229 tumour cells, a finding that supports earlier reports that externally applied inositol phosphates can—perhaps surprisingly—enter into cells. Close examination of the process of FAM‐InsP₅ uptake suggests a mechanism of non‐receptor‐mediated endocytosis, which is blocked at 4 °C and probably involves interaction of the ligand with the glycocalyx. However, our results are difficult to reconcile with antiproliferative mechanisms that require direct interactions of externally applied InsP₅ or InsP₆ with cytosolic proteins, because internalised FAM‐InsP₅ appears in lysosomes and apparently does not enter the cytoplasm. Studies using FAM‐InsP₅ are less difficult and time‐consuming than experiments using InsP₅ or InsP₆, a factor that allowed us to analyse cellular uptake across a range of human cell types, identifying strong cell‐specific differences.     

Nanoscale FeS2 (Pyrite) as a Sustainable Thermoelectric Material

We have synthesized undoped, Co-doped (up to 5%), and Se-doped (up to 4%) FeS₂ materials by mechanical alloying in a planetary ball mill and investigated their thermoelectric properties from room temperature (RT) to 600 K. With decreasing particle size, the undoped FeS₂ samples showed higher electrical conductivity, from 0.02 S cm^?1 for particles with 70 nm grain size up to 3.1 S cm^?1 for the sample with grain size of 16 nm. The Seebeck coefficient of the undoped samples showed a decrease with further grinding, from 128 μV K^?1 at RT for the sample with 70-nm grains down to 101 μV K^?1 for the sample with grain size of 16 nm. The thermal conductivity of the 16-nm undoped sample lay within the range from 1.3 W m^?1 K^?1 at RT to a minimal value of 1.2 W m^?1 K^?1 at 600 K. All doped samples showed improved thermoelectric behavior at 600 K compared with the undoped sample with 16 nm particle size. Cobalt doping modified the p-type semiconducting behavior to n-type and increased the thermal conductivity (2.1 W m^?1 K^?1) but improved the electrical conductivity (41 S cm^?1) and Seebeck coefficient (-129 μV K^?1). Isovalent selenium doping led to a slightly higher thermal conductivity (1.7 W m^?1 K^?1) as well as to an improved electrical conductivity (26 S cm^?1) and Seebeck coefficient (110 μV K^?1). The ZT value of FeS₂ was increased by a factor of five by Co doping and by a factor of three by Se doping.