Digestion rate of legume carbohydrates and glycemic index of legume-based meals.
A study was performed to examine the rate of digestion of available carbohydrate in legumes and its mixtures with cereals, prepared as commonly eaten. The legumes and cereals studied were lentil (Lens sculenta), pea (Pisum sativum), bean (Phaseolus vulgaris, var tortola), rice (Oryza sativa) and spaghetti. Foods were purchased at the city market. Total starch content and the carbohydrate digestion rates were determined using the enzymatic method proposed by Englyst et al. Total starch levels ranged from 7.78 g/100 g in cooked flour bean to 20.6 g/100 g in a bean-spaghetti dish, and dietary fiber contents ranged from 2.4 g/100 g in a cooked 70:30 lentil-rice mixture to 5.26 g/100 g in a cooked whole bean. The rapid digestion rate carbohydrates showed values from 4.8 in the bean soup to 8.9 in the bean-spaghetti combination. The same results show, expressed as rapid available glucose (RAG), the amount of rapid carbohydrate/100 g food or meal as eaten, and as the starch digestion index (SDI), the percentage of rapid carbohydrate digestion rate in relation to the total amount of carbohydrate. The RAG values ranged between 5.0 for cooked beans and 10 for cooked beans and spaghetti, and the SDI ranged between 40 for cooked pea flour and 62 for cooked bean flour. Legumes prepared as soup showed a higher rapid digestion rate than legumes prepared as whole grain. The bean-spaghetti based-meal and the lentil-based meal showed glycemic index mean and standard deviation values of 76.8 +/- 43.4 and 49.3 +/- 29.5, RAG values of 7.0 and 6.0, and SDI values of 57 and 54, respectively. The knowledge of the importance of the carbohydrate digestion rates in human health in increasing, and probably will soon be used in the development of the food pyramid. The foods with a moderate fraction of rapid digestion rate, such as legumes, should be included in the base of the pyramid.


[Fish assembles in the coastal water of Lüsi fishing ground during spring and summer].
Based on data from bottom trawl surveys during spring and summer of 2010, the species composition and community structure of fish community in Lüsi fishing ground were investigated and compared with those at different latitudes. A total of 21 were identified in spring and 28 identified in summer. In spring, it was mainly composed of warm-temperate fish (9 species), accounting for 45.2% and 72.9% in the individual density and biomass density, respectively. Warm-temperate species Larimichthys polyactis, Cynoglossus joyneri and Cynoglossus semilaevis were dominant in spring. In summer, warm water species appeared in large numbers by 21, accounting for 75.2% and 71.7% in the individual density and biomass density, respectively. Collichthys lucidus and Coilia mystus were the predominant species in summer. One way-ANOVA indicated that the temperature and salinity between spring and summer in Lüsi fishing ground were significantly different, which resulted in the succession of fish community, i. e. the gradual decrease of the warm-temperate, neritic and nearshore species and the concomitant gradual increase of warm-water and estuarine species.


Accomodative defocus does not limit development of acuity in infant Macaca nemestrina monkeys.
In an experiment with ten macaque monkeys (Macaca nemestrina), a combination of photorefraction and corneal reflex photography was used to measure simultaneously the plane of focus and direction of gaze while they were presented with fixation targets. The monkeys ranged in age from 2 days to 10 weeks. Some of the infants that were less than 1 month old failed to change accommodation to targets at any distance, whereas others showed limited accommodative abilities. The magnitude of the accommodative response of infants older than 4 weeks appeared to be adultlike. Infant monkey's visual acuity improves dramatically after 4 weeks. These results, which show that the improvement in spatial resolution cannot be accounted for by increased accommodative accuracy, parallel those obtained from human infants where accommodative competence is attained by about 4 months of age.


Globally-Aware Multiple Instance Classifier for Breast Cancer Screening.
Deep learning models designed for visual classification tasks on natural images have become prevalent in medical image analysis. However, medical images differ from typical natural images in many ways, such as significantly higher resolutions and smaller regions of interest. Moreover, both the global structure and local details play important roles in medical image analysis tasks. To address these unique properties of medical images, we propose a neural network that is able to classify breast cancer lesions utilizing information from both a global saliency map and multiple local patches. The proposed model outperforms the ResNet-based baseline and achieves radiologist-level performance in the interpretation of screening mammography. Although our model is trained only with image-level labels, it is able to generate pixel-level saliency maps that provide localization of possible malignant findings.


[Diseases and their treatment in Plato's depiction on the background of views of Hippocratic physicians].
Conception of biological nature of a man included in "Timajos" did not limit itself to presentation of the outline of anatomy and of physiology, but also touched on problems connected with origin of diseases and their treatment. Explanation of mentioned issues was based on the idea of a man as an element of the universe. Plato subscribed opinion that body as all the universe, consisted of four basic primeval elements. Excess or shortage of one of them, exchange of place with other elements or taking over another species property caused disturbances inside the organism leading to diseases. Plato attached large attention to duration of disease symptoms, especially fever. In the dialogue he did not pass over mental diseases, which he treated as illnesses of the soul. Besides he emphasised relationship between somatic and psychical diseases. Plato's opinions reflect the whole Greek natural knowledge, but in some questions they contradict opinions of hippocratic physicians.


Evaluation of the dietary intake data coding process in a clinical setting: Implications for research practice.
High quality dietary intake data is required to support evidence of diet-disease relationships exposed in clinical research. Source data verification may be a useful quality assurance method in this setting. The present pilot study aimed to apply source data verification to evaluate the quality of the data coding process for dietary intake in a clinical trial and to explore potential barriers to data quality in this setting. Using a sample of 20 cases from a clinical trial, source data verification was conducted between three sets of data derived documents: transcripts of audio-recorded diet history interviews, matched paper-based diet history forms and outputs from nutrition analysis software. The number of cases and rates of discrepancies between documents were calculated. A total of five in-depth interviews with dietitians collecting and coding dietary data were thematically analysed. Some 2024 discrepancies were identified. The highest discrepancy rate was 57.49%, and occurred between diet history interviews and nutrition analysis software outputs. Sources of the discrepancies included both quantities and frequencies of food intake. The highest discrepancy rate was for the food group "vegetable products and dishes". In-depth interviews implicated recall bias of trial participants as a cause of discrepancies, but dietitians also acknowledged a possible subconscious influence of having to code reported foods into nutrition analysis software programs. The accuracy of dietary intake data appeared to depend on the level of detailed food data required. More support for participants on reporting consumption, and incorporating supportive tools to guide estimates of food quantities may facilitate a more consistent coding process and improve data quality. This pilot study offers a novel method and an overview of dietary intake data coding measurement errors. These findings may warrant further investigation in a larger sample.


Determinants of the treatment climate in psychiatric and substance abuse programs: implications for improving patient outcomes.
This study examined determinants of the treatment climate in 89 psychiatric and substance abuse programs. Clearer policies giving patients more control, and more health and treatment services, were related to more supportive, autonomous, expressive, and practically oriented milieus. In contrast, more policy choice and daily living assistance were associated with less support, personal expression, and practical orientation. Programs in which staff had more paraprofessionals and better team functioning, and in which patients had more social resources and better mental functioning, tended to be more supportive and to have more emphasis on autonomy, personal expression, and practical orientation. Based on these results, we suggest some guidelines on how to change the treatment milieu to benefit patients' adjustment.


Rosuvastatin, a new HMG-CoA reductase inhibitor, protects ischemic reperfused myocardium in normocholesterolemic rats.
The 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) have been shown to upregulate endothelial nitric oxide synthase in isolated endothelial cells in a manner that is independent of their lipid-lowering effects. Nitric oxide inhibits polymorphonuclear leukocyte (PMN) adherence and attenuates cardiac dysfunction caused by PMNs after ischemia/reperfusion. Therefore, the authors hypothesized that a new statin, rosuvastatin, could attenuate PMN-induced cardiac dysfunction, and examined the effects of rosuvastatin in isolated ischemic (20 min) and reperfused (45 min) rat hearts perfused with PMNs. Rosuvastatin (0.25 or 1.25 mg/kg) given 18 h before ischemia/reperfusion significantly improved left ventricular developed pressure (P < 0.01) and the maximal rate of development of left ventricular developed pressure (+dP/dt(max), P < 0.01) compared with ischemia/reperfused hearts obtained from rats given 0.9% NaCl. The time point for the improved cardiac performance caused by rosuvastatin (1.25 mg/kg) was 20 min after reperfusion. In addition, rosuvastatin significantly reduced PMN adherence to the vascular endothelium and subsequent infiltration into the postischemic myocardium (P < 0.01). The nitric oxide synthase inhibitor N omega-nitro-l-arginine methyl ester (50 micromol/l) blocked these cardioprotective effects. These results provide evidence that rosuvastatin significantly attenuates PMN-induced cardiac contractile dysfunction in the isolated perfused rat heart.


The JAZ family of repressors is the missing link in jasmonate signalling.
Jasmonates are essential phytohormones for plant development and survival. However, the molecular details of their signalling pathway remain largely unknown. The identification more than a decade ago of COI1 as an F-box protein suggested the existence of a repressor of jasmonate responses that is targeted by the SCF(COI1) complex for proteasome degradation in response to jasmonate. Here we report the identification of JASMONATE-INSENSITIVE 3 (JAI3) and a family of related proteins named JAZ (jasmonate ZIM-domain), in Arabidopsis thaliana. Our results demonstrate that JAI3 and other JAZs are direct targets of the SCF(COI1) E3 ubiquitin ligase and jasmonate treatment induces their proteasome degradation. Moreover, JAI3 negatively regulates the key transcriptional activator of jasmonate responses, MYC2. The JAZ family therefore represents the molecular link between the two previously known steps in the jasmonate pathway. Furthermore, we demonstrate the existence of a regulatory feed-back loop involving MYC2 and JAZ proteins, which provides a mechanistic explanation for the pulsed response to jasmonate and the subsequent desensitization of the cell.


Algorithm for navigated ESS.
ENT navigation has given new opportunities in performing Endoscopic Sinus Surgery (ESS) and improving surgical outcome of the patients` treatment. ESS assisted by a navigation system could be called Navigated Endoscopic Sinus Surgery (NESS). As it is generally accepted that the NESS should be performed only in cases of complex anatomy and pathology, it has not yet been established as a state-of-the-art procedure and thus not used on a daily basis. This paper presents an algorithm for use of a navigation system for basic ESS in the treatment of chronic rhinosinusitis (CRS). The algorithm includes five units that should be highlighted using a navigation system. They are as follows: 1) nasal vestibule unit, 2) OMC unit, 3) anterior ethmoid unit, 4) posterior ethmoid unit, and 5) sphenoid unit. Each unit has a shape of a triangular pyramid and consists of at least four reference points or landmarks. As many landmarks as possible should be marked when determining one of the five units. Navigated orientation in each unit should always precede any surgical intervention. The algorithm should improve the learning curve of trainees and enable surgeons to use the navigation system routinely and systematically.


[Follow-up of patients of the Department of Psychotherapy and Psychosomatic Medicine of the Psychiatric Clinic of the University of Munich. Attempt at an evaluation of the effect of psychotherapeutic counseling].
In order to find out the patients reaction to psychotherapeutic counselling and to see if a psychotherapy suggested by the counselling therapist was accepted by the patients, a questionnaire survey was made. 71.3% of the questionnaires were answered. Half of the patients had followed the advice and began a psychotherapy. The subjective reaction to counseling was equally positive, rather positive and negative. These results are often found in psychotherapeutic follow-up studies. In accordance with literature it was also found that neurotic patients in contrast to those with mainly psychosomatic and psychovegetative complaints, younger and middle class patients, women as well as patients coming spontaneously were more likely to undergo psychotherapeutic treatment than their comparison groups. Further it was found: Men were more often advised to begin a behaviour-therapy, women to undergo psychoanalysis. Patients who had been treated in the University Psychiatric Hospital were more likely to follow the advice given by the counselling therapist than those patients being sent by other hospitals or general practitioners. If the counselling therapist provided a possibility where his client could begin with psychotherapy immediately, the client was more likely to undergo therapy than if he had to arrange for psychotherapeutic treatment mainly by himself.


Synergistic antitumor effect of 5-fluorouracil and withaferin-A induces endoplasmic reticulum stress-mediated autophagy and apoptosis in colorectal cancer cells.
The development of chemo-resistance against 5-fluorouracil (5-FU) in tumor cells is one of the main debacles in colorectal cancer (CRC) patients. A recent combination of 5-FU with oxaliplatin or cetuximab drastically improves the survival rate in CRC patients; however, the toxicity issue cannot be evaded completely. Thus, searching for novel drug combinations with high specificity and low toxicity is seemingly important. Owing to the less undesirable effects of natural products on normal cells, here we investigated the synergistic antitumor effect of withaferin-A (WA) in combination with 5-FU. Our results demonstrate that the combination of WA and 5-FU induces a significant antiproliferative effect and modulates endoplasmic reticulum (ER) stress in favor of cell death in colorectal cancer (CRC) cells. Mechanistically, the combination upregulates the expression of ER stress sensors (BiP, PERK, CHOP, ATF-4, and eIF2α) and executes PERK axis mediated apoptosis in CRC cells. Additionally, the combined treatment of WA and 5-FU mediated ER stress induces autophagy and apoptosis, which were confirmed by immunoblotting, acridine orange (AO) staining and annexin-V FITC by flow cytometry. In contrast, inhibition of ER stress with salubrinal significantly decreases both autophagic and apoptotic cell populations. Moreover, pharmacological inhibition of either autophagy or apoptosis by their respective inhibitors 3-methyladenine (3-MA) or carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoro-methyl ketone (Z-VAD-FMK) decreases their respective population of cells but could not affect either of the population significantly. Finally, the combination attenuates the expression of β-catenin pathway associated proteins and arrests cell cycle at the G2M phase in CRC cells. In summary, the combination of WA and 5-FU decreases cell viability by inducing ER stress-mediated induction of autophagy and apoptosis, inhibiting the β-catenin pathway and arresting the cell cycle at a G2M phase in CRC cells.


Interleukin-16 promotes cardiac fibrosis and myocardial stiffening in heart failure with preserved ejection fraction.
Chronic heart failure (CHF) with preserved left ventricular (LV) ejection fraction (HFpEF) is observed in half of all patients with CHF and carries the same poor prognosis as CHF with reduced LV ejection fraction (HFrEF). In contrast to HFrEF, there is no established therapy for HFpEF. Chronic inflammation contributes to cardiac fibrosis, a crucial factor in HFpEF; however, inflammatory mechanisms and mediators involved in the development of HFpEF remain unclear. Therefore, we sought to identify novel inflammatory mediators involved in this process. An analysis by multiplex-bead array assay revealed that serum interleukin-16 (IL-16) levels were specifically elevated in patients with HFpEF compared with HFrEF and controls. This was confirmed by enzyme-linked immunosorbent assay in HFpEF patients and controls, and serum IL-16 levels showed a significant association with indices of LV diastolic dysfunction. Serum IL-16 levels were also elevated in a rat model of HFpEF and positively correlated with LV end-diastolic pressure, lung weight and LV myocardial stiffness constant. The cardiac expression of IL-16 was upregulated in the HFpEF rat model. Enhanced cardiac expression of IL-16 in transgenic mice induced cardiac fibrosis and LV myocardial stiffening accompanied by increased macrophage infiltration. Treatment with anti-IL-16 neutralizing antibody ameliorated cardiac fibrosis in the mouse model of angiotensin II-induced hypertension. Our data indicate that IL-16 is a mediator of LV myocardial fibrosis and stiffening in HFpEF, and that the blockade of IL-16 could be a possible therapeutic option for HFpEF.


Effect of echo time pair selection on quantitative analysis for adrenal tumor characterization with in-phase and opposed-phase MR imaging: initial experience.
To determine the effect of two pairs of echo times (TEs) for in-phase (IP) and opposed-phase (OP) 3.0-T magnetic resonance (MR) imaging on (a) quantitative analysis prospectively in a phantom study and (b) diagnostic accuracy retrospectively in a clinical study of adrenal tumors, with use of various reference standards in the clinical study. A fat-saline phantom was used to perform IP and OP 3.0-T MR imaging for various fat fractions. The institutional review board approved this HIPAA-compliant study, with waiver of informed consent. Single-breath-hold IP and OP 3.0-T MR images in 21 patients (14 women, seven men; mean age, 63 years) with 23 adrenal tumors (16 adenomas, six metastases, one adrenocortical carcinoma) were reviewed. The MR protocol involved two acquisition schemes: In scheme A, the first OP echo (approximately 1.5-msec TE) and the second IP echo (approximately 4.9-msec TE) were acquired. In scheme B, the first IP echo (approximately 2.4-msec TE) and the third OP echo (approximately 5.8-msec TE) were acquired. Quantitative analysis was performed, and analysis of variance was used to test for differences between adenomas and nonadenomas. In the phantom study, scheme B did not enable discrimination among voxels that had small amounts of fat. In the clinical study, no overlap in signal intensity (SI) index values between adenomas and nonadenomas was seen (P < .05) with scheme A. However, with scheme B, no overlap in the adrenal gland SI-to-liver SI ratio between adenomas and nonadenomas was seen (P < .05). With scheme B, no overlap in adrenal gland SI index-to-liver SI index ratio between adenomas and nonadenomas was seen (P < .05). This initial experience indicates SI index is the most reliable parameter for characterization of adrenal tumors with 3.0-T MR imaging when obtaining OP echo before IP echo. When acquiring IP echo before OP echo, however, nonadenomas can be mistaken as adenomas with use of the SI index value.


H4K16ac activates the transcription of transposable elements and contributes to their cis-regulatory function.
Mammalian genomes harbor abundant transposable elements (TEs) and their remnants, with numerous epigenetic repression mechanisms enacted to silence TE transcription. However, TEs are upregulated during early development, neuronal lineage, and cancers, although the epigenetic factors contributing to the transcription of TEs have yet to be fully elucidated. Here, we demonstrate that the male-specific lethal (MSL)-complex-mediated histone H4 acetylation at lysine 16 (H4K16ac) is enriched at TEs in human embryonic stem cells (hESCs) and cancer cells. This in turn activates transcription of subsets of full-length long interspersed nuclear elements (LINE1s, L1s) and endogenous retrovirus (ERV) long terminal repeats (LTRs). Furthermore, we show that the H4K16ac-marked L1 and LTR subfamilies display enhancer-like functions and are enriched in genomic locations with chromatin features associated with active enhancers. Importantly, such regions often reside at boundaries of topologically associated domains and loop with genes. CRISPR-based epigenetic perturbation and genetic deletion of L1s reveal that H4K16ac-marked L1s and LTRs regulate the expression of genes in cis. Overall, TEs enriched with H4K16ac contribute to the cis-regulatory landscape at specific genomic locations by maintaining an active chromatin landscape at TEs.


Social influence and individual risk factors of HIV unsafe sex among female entertainment workers in China.
Female entertainment workers in China are at increased sexual risk of HIV, but causes of their unprotected sex remain poorly understood. We develop a model that integrates information-motivation-behavioral skills (IMB) with social influences and test the model in a venue-based sample of 732 female entertainment workers in Shanghai. Most IMB and social influence measures are statistically significant in bivariate relationships to condom use; only HIV prevention motivation and behavioral self-efficacy remain significant in the multiple regressions. Self-efficacy in condom use is the most proximate correlate, mediating the relationship between information and motivation and condom use. Both peer and venue supports are important, but their influences over condom use are indirect and mediated through prevention motivation and/or self-efficacy. Behavioral intervention is urgently needed and should take a multilevel approach, emphasizing behavioral skills training and promoting a supportive social/working environment.


A novel tissue-based ß-catenin gene and immunohistochemical analysis to exclude familial adenomatous polyposis among children with hepatoblastoma tumors.
The Wnt/β-catenin pathway plays a central role in the pathogenesis of most hepatoblastomas (HBs), that is, up to 60-80% carry activating CTNNB1 mutations. HBs can however also be the first manifestation of familial adenomatous polyposis (FAP). As this is a severe disease, it is important for the patient and related family members to firmly exclude FAP at an early stage. Current diagnosis largely depends on APC germline mutation detection on genomic DNA, which is associated with 10-20% false-negative results. Here, we establish and validate a tissue-based β-catenin gene and immunohistochemical analysis, which complements germline mutation screening to exclude the diagnosis of FAP among HB patients. Tumor tissues of 18 HB patients, including three FAP cases were subjected to CTNNB1 exon 3 mutational analysis and immunohistochemistry comparing staining patterns for total and exon 3 specific β-catenin antibodies. Our novel tissue-based method reliably identified all three FAP patients. Their tumors were characterized by a wild-type exon 3 sequence and a comparable nuclear staining for both antibodies. In contrast, the non-FAP tumors carried missense CTNNB1 mutations combined with a clearly reduced staining for the exon 3 antibody, or complete loss of staining in case of lesions with exon 3 deletions. We have successfully established and validated a novel ß-catenin gene and immunohistochemical diagnostic method, which, when combined with routine germline DNA testing, allows the exclusion of the diagnosis of FAP among HB patients.


Traditional chinese medicine-based network pharmacology could lead to new multicompound drug discovery.
Current strategies for drug discovery have reached a bottleneck where the paradigm is generally "one gene, one drug, one disease." However, using holistic and systemic views, network pharmacology may be the next paradigm in drug discovery. Based on network pharmacology, a combinational drug with two or more compounds could offer beneficial synergistic effects for complex diseases. Interestingly, traditional chinese medicine (TCM) has been practicing holistic views for over 3,000 years, and its distinguished feature is using herbal formulas to treat diseases based on the unique pattern classification. Though TCM herbal formulas are acknowledged as a great source for drug discovery, no drug discovery strategies compatible with the multidimensional complexities of TCM herbal formulas have been developed. In this paper, we highlighted some novel paradigms in TCM-based network pharmacology and new drug discovery. A multiple compound drug can be discovered by merging herbal formula-based pharmacological networks with TCM pattern-based disease molecular networks. Herbal formulas would be a source for multiple compound drug candidates, and the TCM pattern in the disease would be an indication for a new drug.


Penetrating cardiac injury.
It is understood that penetrating cardiac trauma is a highly lethal injury and those surviving to hospital have an overall mortality approaching 80%. Reported mortality figures vary widely and are extremely dependent on mechanism of wounding, cardiac chambers involved and possibly the presence of cardiac tamponade. Despite significant advances in prehospital care, operative techniques, and intensive care management, the mortality has not changed over several decades. This article will review the anatomic regions of concern for a cardiac injury, clinical presentation, and physical findings. The need for an expeditious evaluation and modalities available including, plain radiographs, sub-xiphoid window, and echocardiography will be considered. Options for surgical exposure, technical details of repairing cardiac injuries, and special circumstances such as injury adjacent to a coronary artery and intra-cardiac shunts are discussed in detail. Outcome data and future directions in managing this challenging injury are also examined.


Revisiting PFA-mediated tissue fixation chemistry: FixEL enables trapping of small molecules in the brain to visualize their distribution changes.
Various small molecules have been used as functional probes for tissue imaging in medical diagnosis and pharmaceutical drugs for disease treatment. The spatial distribution, target selectivity, and diffusion/excretion kinetics of small molecules in structurally complicated specimens are critical for function. However, robust methods for precisely evaluating these parameters in the brain have been limited. Herein, we report a new method termed "fixation-driven chemical cross-linking of exogenous ligands (FixEL)," which traps and images exogenously administered molecules of interest (MOIs) in complex tissues. This method relies on protein-MOI interactions and chemical cross-linking of amine-tethered MOI with paraformaldehyde used for perfusion fixation. FixEL is used to obtain images of the distribution of the small molecules, which addresses selective/nonselective binding to proteins, time-dependent localization changes, and diffusion/retention kinetics of MOIs such as the scaffold of PET tracer derivatives or drug-like small molecules.


Lymphocyte Populations in the Adventitial Layer of Hydatid Cysts in Cattle: Relationship With Cyst Fertility Status and Fasciola Hepatica Co-Infection.
Cystic echinococcosis is a worldwide zoonosis caused by the cestode Echinococcus granulosus. Two types of hydatid cysts occur in intermediate hosts: fertile cysts that generate protoscoleces from the germinal layer of the cyst, and infertile cysts that do not produce protoscoleces and are unable to continue the life cycle of the parasite. The adventitial layer, a host-derived fibrous capsule surrounding the hydatid cyst, is suggested to play an important role in local immune regulation during infection and in fertility of the cysts. Fasciola hepatica, another important parasite of cattle, induces a characteristic Th2-like immune response that could modulate the immune response against E. granulosus. Natural co-infection of both parasites is common in cattle, but no reports describe the local immune response against E. granulosus with F. hepatica infection in the same host. This study analyzed the number and distribution of T and B cells in the adventitial layer of liver and lung cysts and the relationship with cyst fertility and F. hepatica co-infection. T lymphocytes were the predominant cell type in the adventitial layer of infertile hydatid cysts and were more numerous in infertile hydatid cysts. B lymphocyte numbers were not associated with hydatid cyst fertility. Mast cells were infrequent in the adventitial layer. The number of T and B cells was not associated with F. hepatica co-infection. The present study contributes to the understanding of local immune responses in bovine cystic echinococcosis.


Feedback modulation of neural network synchrony and seizure susceptibility by Mdm2-p53-Nedd4-2 signaling.
Neural network synchrony is a critical factor in regulating information transmission through the nervous system. Improperly regulated neural network synchrony is implicated in pathophysiological conditions such as epilepsy. Despite the awareness of its importance, the molecular signaling underlying the regulation of neural network synchrony, especially after stimulation, remains largely unknown. In this study, we show that elevation of neuronal activity by the GABA(A) receptor antagonist, Picrotoxin, increases neural network synchrony in primary mouse cortical neuron cultures. The elevation of neuronal activity triggers Mdm2-dependent degradation of the tumor suppressor p53. We show here that blocking the degradation of p53 further enhances Picrotoxin-induced neural network synchrony, while promoting the inhibition of p53 with a p53 inhibitor reduces Picrotoxin-induced neural network synchrony. These data suggest that Mdm2-p53 signaling mediates a feedback mechanism to fine-tune neural network synchrony after activity stimulation. Furthermore, genetically reducing the expression of a direct target gene of p53, Nedd4-2, elevates neural network synchrony basally and occludes the effect of Picrotoxin. Finally, using a kainic acid-induced seizure model in mice, we show that alterations of Mdm2-p53-Nedd4-2 signaling affect seizure susceptibility. Together, our findings elucidate a critical role of Mdm2-p53-Nedd4-2 signaling underlying the regulation of neural network synchrony and seizure susceptibility and reveal potential therapeutic targets for hyperexcitability-associated neurological disorders.


Different forms of the bovine PrP gene have five or six copies of a short, G-C-rich element within the protein-coding exon.
Current models of the virus-like agents of scrapie and bovine spongiform encephalopathy (BSE) have to take into account that structural changes in a host-encoded protein (PrP protein) exhibit an effect on the time course of these diseases and the survival time of any man or animal exposed to these pathogens. We report here the sequence of different forms of the bovine PrP gene which contain either five or six copies of a short, G-C-rich element which encodes the octapeptide Pro-His-Gly-Gly-Gly-Trp-Gly-Gln or its longer variants Pro-Gln/His-Gly-Gly-Gly-Gly-Trp-Gly-Gln. Out of 12 cattle, we found eight animals homozygous for genes with six copies of the Gly-rich peptide (6:6), while four were heterozygous (6:5). Two confirmed cases of BSE occurred in (6:6) homozygous animals.


Sex differences in myosin heavy chain isoforms of human failing and nonfailing atria.
Mammalian hearts express two myosin heavy chain (MHC) isoforms, which drive contractions with different kinetics and power-generating ability. The expression of the isoform that is associated with more rapid contraction kinetics and greater power output, MHC-α, is downregulated, with a concurrent increase in the relative amount of the slower isoform, MHC-β, during the progression to experimentally induced or disease-related heart failure. This change in protein expression has been well studied in right and left ventricles in heart failure models and in humans with failure. Relatively little quantitative data exists regarding MHC isoform expression shifts in human failing atria. We previously reported significant increases in the relative amount of MHC-β in the human failing left atrium. The results of that study suggested that there might be a sex-related difference in the level of MHC-β in the left atrium, but the number of female subjects was insufficient for statistical analysis. The objective of this study was to test whether there is, in fact, a sex-related difference in the level of MHC-β in the right and left atria of humans with cardiomyopathy. The results indicate that significant differences exist in atrial MHC isoform expression between men and women who are in failure. The results also revealed an unexpected twofold greater amount of MHC-β in the nonfailing left atrium of women, compared with men. The observed sex-related differences in MHC isoform expression could impact ventricular diastolic filling during normal daily activities, as well as during physiologically stressful events.


Rapid Actuation of Thermo-Responsive Polymer Networks: Investigation of the Transition Kinetics.
The swelling and collapsing of thermo-responsive poly(N-isopropylacrylamide)-based polymer (pNIPAAm) networks are investigated in order to reveal the dependency on their kinetics and maximum possible actuation speed. The pNIPAAm-based network was attached as thin hydrogel film to lithographically prepared gold nanoparticle arrays to exploit their localized surface plasmon resonance (LSPR) for rapid local heating. The same substrate also served for LSPR-based monitoring of the reversible collapsing and swelling of the pNIPAAm network through its pronounced refractive index changes. The obtained data reveal signatures of multiple phases during the volume transition, which are driven by the diffusion of water molecules into and out of the network structure and by polymer chain re-arrangement. For the micrometer-thick hydrogel film in the swollen state, the layer can respond as fast as several milliseconds depending on the strength of the heating optical pulse and on the tuning of the ambient temperature with respect to the lower critical solution temperature of the polymer. Distinct differences in the time constants of swelling and collapse are observed and attributed to the dependence of the cooperative diffusion coefficient of polymer chains on polymer volume fraction. The reported results may provide guidelines for novel miniature actuator designs and micromachines that take advantages of the non-reciprocal temperature-induced volume transitions in thermo-responsive hydrogel materials.


High Prevalence of Buprenorphine in Prenatal Drug Screens in an Appalachian City.
To define the magnitude of buprenorphine presence in the urine drug screens of pregnant women and to assess the presence of illicit buprenorphine use versus the presence of prescribed buprenorphine use. Initial prenatal drug screen results for all pregnant patients in our practice for a 1-year period were analyzed and tabulated. Buprenorphine was found in the urine drug screens of 16% of pregnant patients. The presence of buprenorphine was by far the highest for any substance associated with neonatal abstinence syndrome (NAS). We estimate that the exposure to buprenorphine of approximately one-third of individuals in our population is associated with illicit buprenorphine use. The high rate of NAS in our region is primarily associated with both illicit and prescribed buprenorphine rather than other substances. Buprenorphine usage at the time that prenatal care is initiated, rather than opiate use at the onset of prenatal care, is the underlying factor that must be addressed if our region is to successfully combat our high rates of NAS.


Hanging by a thread? Forests and drought.
Trees are the living foundations on which most terrestrial biodiversity is built. Central to the success of trees are their woody bodies, which connect their elevated photosynthetic canopies with the essential belowground activities of water and nutrient acquisition. The slow construction of these carbon-dense, woody skeletons leads to a slow generation time, leaving trees and forests highly susceptible to rapid changes in climate. Other long-lived, sessile organisms such as corals appear to be poorly equipped to survive rapid changes, which raises questions about the vulnerability of contemporary forests to future climate change. The emerging view that, similar to corals, tree species have rather inflexible damage thresholds, particularly in terms of water stress, is especially concerning. This Review examines recent progress in our understanding of how the future looks for forests growing in a hotter and drier atmosphere.


Effects of the 'fusion peptide' from measles virus on the structure of N-methyl dioleoylphosphatidylethanolamine membranes and their fusion with Sendai virus.
31P nuclear magnetic resonance spectroscopy (31P-NMR) was used to study phospholipid organization in hydrated preparations of N-methyl dioleoylphosphatidylethanolamine and a 'fusion peptide' with the sequence: FAGV-VLAGAALGVAAAAQI, which corresponds to the amino terminus of the F1 subunit of the membrane fusion protein of measles virus. These amino acids are believed to mediate syncytia formation, host-cell penetration and hemolysis by infectious virus. The presence of the peptide at 0.5 mole percent significantly facilitated the formation of isotropic 31P resonances. The effects at 1 mole percent peptide were substantially enhanced over the effects observed at 0.5 mole percent, leading to a decrease in the onset temperature of the formation of the isotropic 31P-NMR resonances by about 30 degrees C. The formation of such isotropic 31P-NMR resonances has been previously associated with an increased rate of fusion of large unilamellar vesicles composed of N-methyl dioleoylphosphatidylethanolamine. Enhanced fusion of octadecyl rhodamine-labelled Sendai virus with N-methyl dioleoylphosphatidylethanolamine large unilamellar vesicles was observed when the 'fusion peptide' was incorporated into the large unilamellar vesicles.


Synthesis and biological evaluation of some new amide moiety bearing quinoxaline derivatives as antimicrobial agents.
In this study, we aimed to synthesize some new quinoxaline derivatives bearing amide moiety and to evaluate their antimicrobial activity. A set of 16 novel compounds of N-[2,3-bis(4-methoxy/methylphenyl)quinoxalin-6-yl]-substituted benzamide derivatives were synthesized by reacting 2,3-bis(4-methoxyphenyl)-6-aminoquinoxaline or 2,3-bis(4-methylphenyl)-6-aminoquinoxaline with benzoyl chloride derivatives in tetrahydrofuran and investigated for their antimicrobial activity. The structures of the obtained final compounds were confirmed by spectral data (IR, (1)H-NMR, (13)C-NMR and MS). The antimicrobial activity of the compounds were determined by using the microbroth dilution method. Antimicrobial activity results revealed that synthesized compounds exhibited remarkable activity against Candida krusei (ATCC 6258) and Candida parapsilosis (ATCC 22019).


PP13 and PAPP-A in the First and Second Trimesters: Predictive Factors for Preeclampsia?
Background. Preeclampsia affects 5-6% of all pregnancies. Predictive factors of preeclampsia can be helpful in early diagnosis of this disease. In this study the predictive values of biochemical markers placenta protein 13 (PP13) and pregnancy-associated plasma protein A (PAPP-A) have been assessed in early diagnosis of preeclampsia. Methods. This case-control study was conducted on 1500 women who presented to a healthcare center of Sari, Iran, between 2010 and 2011. Blood samples were drawn in weeks 11-13 and 24-28 of pregnancy. Of them who developed preeclampsia were considered as case group. A control group consisted of similar women regarding mean age, body mass index (BMI), and pregnancy age. PAPP-A and PP13 serum levels were measured. Data were analyzed using proper statistical tests. Results. PAPP-A and PP13 serum levels were significantly lower in both the first and second trimesters in women who developed preeclampsia (P < 0.001). The cumulative value of all four variables with cut-off point of 238.5 has sensitivity, specificity of 91.0%, and undercurve surface of 0.968 which is the most diagnostic value for preeclampsia. Conclusion. It is possible to advantage measuring of PAPP-A and PP13 in the first and second trimesters especially their cumulative values in both trimesters for prediction of the incidence of preeclampsia.


The purpose, content and development phases of a graduate entry ePortfolio in occupational therapy.
There is a trend in higher education towards the use of digital or electronic portfolios (ePortfolios) to collect evidence that demonstrates learning and skill development. There are very few papers that examine the key features and what to include in an occupational therapy ePortfolio. This study presents an approach to developing a graduate entry ePortfolio to prepare occupational therapy students for transition to work. An e-Delphi approach was used to gather the opinions of eight categories of experts. Three rounds of questionnaires were used to explore the purpose, terminology, content, structure, and development phases of a graduate entry ePortfolio. Key stakeholders indicated that the purpose of a professional portfolio is to create a professional profile, record experiences, skills and behaviours, and promote recording of lifelong learning and achievements. Delphi expert panel members emphasised the importance of personal choice in selecting evidence recorded in each collection, which must be guided by ethical decision-making. The findings of the study are important to students who wish to build an ePortfolio to organise and display evidence of competence prior to graduation in readiness to enter the workforce. The recommendations also will be of value to occupational therapy educators in curriculum development.


ICM conversion to epiblast by FGF/ERK inhibition is limited in time and requires transcription and protein degradation.
Inner cell Mass (ICM) specification into epiblast (Epi) and primitive endoderm (PrE) is an asynchronous and progressive process taking place between E3.0 to E3.75 under the control of the Fibroblast Growth Factor (FGF)/Extracellular signal-Regulated Kinase (ERK) signaling pathway. Here, we have analyzed in details the kinetics of specification and found that ICM cell responsiveness to the up and down regulation of FGF signaling activity are temporally distinct. We also showed that PrE progenitors are generated later than Epi progenitors. We further demonstrated that, during this late phase of specification, a 4 hours period of FGF/ERK inhibition prior E3.75 is sufficient to convert ICM cells into Epi. Finally, we showed that ICM conversion into Epi in response to inhibition during this short time window requires both transcription and proteasome degradation. Collectively, our data give new insights into the timing and mechanisms involved in the process of ICM specification.


Socioeconomic status moderates the association between John Henryism and NEO PI-R personality domains.
To investigate associations between John Henryism (JH) and NEO Personality Inventory-Revised (PI-R) personality domains. JH-a strong behavioral predisposition to engage in high-effort coping with difficult psychosocial and economic stressors-has been associated with poor health, particularly among persons in lower socioeconomic (SES) groups. Unfavorable personality profiles have also been frequently linked to poor health; however, no studies have yet examined what global personality traits characterize JH. Hypotheses were examined, using data from a sample of 233 community volunteers (mean age, 33 years; 61% black and 39% white) recruited specifically to represent the full range of the SES gradient. Personality (NEO PI-R) and active coping (12-item JH scale) measures and covariates were derived from baseline interviews. In a multiple regression analysis, independent of SES, JH was positively associated with Conscientiousness (C) (p < .001) and Extraversion (E) (p < .001), whereas the combination of low JH and high SES was associated with Neuroticism (N) (p = .02) When examining associations between JH and combinations of NEO PI-R domains called "styles," high JH was most strongly associated with a high E/high C "Go-Getters" style of activity, whereas low JH was associated with the low E/high Openness (O) "Introspectors" style. In facet level data, the most robust associations with JH were found for five C and five E facets. High JH was associated with higher scores on C and E, but the combination of low JH and high SES was associated with higher scores on N.


Motor neuron-like NSC-34 cells as a new model for the study of vitamin D metabolism in the brain.
Vitamin D3 is a pro-hormone, which is sequentially activated by 25- and 1α-hydroxylation to form 25-hydroxyvitamin D3 [25(OH)D3] and 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D3], respectively. Subsequent inactivation is performed by 24-hydroxylation. These reactions are carried out by a series of CYP450 enzymes. The 25-hydroxylation involves mainly CYP2R1 and CYP27A1, whereas 1α-hydroxylation and 24-hydroxylation are catalyzed by CYP27B1 and CYP24A1, respectively, and are tightly regulated to maintain adequate levels of the active vitamin D hormone, 1α,25(OH)2D3. Altered circulating vitamin D levels, in particular 25(OH)D3, have been linked to several disorders of the nervous system, e.g., schizophrenia and Parkinson disease. However, little is known about the mechanisms of vitamin D actions in the neurons. In this study, we examined vitamin D metabolism and its regulation in a murine motor neuron-like hybrid cell line, NSC-34. We found that these cells express mRNAs for the four major CYP450 enzymes involved in vitamin D activation and inactivation, and vitamin D receptor (VDR) that mediates vitamin D actions. We also found high levels of CYP24A1-dependent 24,25-dihydroxyvitamin D3 [24,25(OH)2D3] production, that was inhibited by the well-known CYP enzyme inhibitor ketoconazole and by several inhibitors that are more specific for CYP24A1. Furthermore, CYP24A1 mRNA levels in NSC-34 cells were up-regulated by 1α,25(OH)2D3 and its synthetic analogs, EB1089 and tacalcitol. Our results suggest that NSC-34 cells could be a novel model for the studies of neuronal vitamin D metabolism and its mechanism of actions.


Inflammatory arthritis in caspase 1 gene-deficient mice: contribution of proteinase 3 to caspase 1-independent production of bioactive interleukin-1beta.
Caspase 1, a known cysteine protease, is a critical component of the inflammasome. Both caspase 1 and neutrophil serine proteases such as proteinase 3 (PR3) can process pro-interleukin-1beta (proIL-1beta), a crucial cytokine linked to the pathogenesis of rheumatoid arthritis. This study was undertaken to establish the relative importance of caspase 1 and serine proteases in mouse models of acute and chronic inflammatory arthritis. Acute and chronic arthritis were induced in caspase 1-/- mice, and the lack of caspase 1 was investigated for its effects on joint swelling, cartilage metabolism, and histopathologic features. In addition, caspase 1 activity was inhibited in mice lacking active cysteine proteases, and the effects of dual blockade of caspase 1 and serine proteases on arthritis severity and histopathologic features were evaluated. Surprisingly, caspase 1-/- mice, in a model of acute (neutrophil-dominated) arthritis, developed joint swelling to an extent similar to that in wild-type control mice. Joint fluid concentrations of bioactive IL-1beta were comparable in caspase 1-/- mice and controls. In contrast, induction of chronic arthritis (characterized by minimal numbers of neutrophils) in caspase 1-/- mice led to reduced joint inflammation and less cartilage damage, implying a caspase 1-dependent role in this process. In mice lacking neutrophil serine PR3, inhibition of caspase 1 activity resulted in decreased bioactive IL-1beta concentrations in the synovial tissue and less suppression of chondrocyte anabolic function. In addition, dual blockade of both PR3 and caspase 1 led to protection against cartilage and bone destruction. Caspase 1 deficiency does not affect neutrophil-dominated joint inflammation, whereas in chronic arthritis, the lack of caspase 1 results in reduced joint inflammation and cartilage destruction. These findings suggest that inhibitors of caspase 1 are not able to interfere with the whole spectrum of IL-1beta production, and therefore such inhibitors may be of therapeutic value only in inflammatory conditions in which limited numbers of neutrophils are present.


Pituitary stem/progenitor cells: embryonic players in the adult gland?
The pituitary gland represents the endocrine core of the body, and its hormonal output governs many key physiological processes. Because endocrine demands frequently change, the pituitary has to flexibly remodel its hormone-producing cell compartment. One mechanism of pituitary plasticity may rely on the generation of new hormonal cells from resident stem/progenitor cells. Existence of such 'master' cells in the pituitary has in the past repeatedly been postulated. Only recently, however, very plausible candidates have been identified that express stem cell-associated markers and signalling factors, and display the stem/progenitor cell characteristics of multipotency, efflux capacity (side population phenotype) and niche-like organization. In other adult tissues, stem cells recapitulate the embryonic developmental path on their course towards mature specialized cells. Interestingly, the pituitary stem/progenitor cell compartment shows prominent expression of transcriptional regulators and signalling factors that play a pivotal role during pituitary embryogenesis. This review summarizes the recent progress in pituitary stem/progenitor cell identification, highlights their potential embryonic phenotype, sketches a tentative stem/progenitor cell model, and discusses further research and challenges. Recognizing and scrutinizing the pituitary stem/progenitor cells as embryonic players in the adult gland may profoundly impact on our still poor understanding of the mechanisms underlying pituitary cell turnover and plasticity.


Nuclear transfer of sand cat cells into enucleated domestic cat oocytes is affected by cryopreservation of donor cells.
In the present study, we used the sand cat (Felis margarita) as a somatic cell donor to evaluate whether cryopreservation of donor cells alters viability and epigenetic events in donor cells and affects in vitro and in vivo developmental competence of derived embryos. In Experiment 1, flow cytometry analysis revealed that the percentage of necrosis and apoptosis in cells analyzed immediately after freezing/thawing (61 vs. 8.1%, respectively) was higher than that observed in frozen/thawed cells cultured for 18 h (6.9 vs. 3.3%, respectively) or 5 days (38 vs. 2.6%; respectively). The relative acetylation level of H3K9 was lower in frozen/thawed cells (5.4%) compared to that found in cultured cells (60.1%). In Experiment 2, embryos reconstructed with frozen/thawed cells had a lower cleavage rate (85%; day 2) than did embryos reconstructed with cultured cells (95%), while development to the blastocyst stage (day 8) was not affected by cell treatment (17.0% with frozen/thawed cells vs. 16.5% with cultured cells). In Experiment 3, pregnancy rates were similar between both cell treatments (32% with frozen/thawed cells vs. 30% with cultured cells), but the number of embryos that were implanted, and the number of fetuses that developed to term was lower for embryos reconstructed with frozen/thawed cells (1.2 and 0.3%, respectively) than those reconstructed with cultured cells (2.6 and 1.8%, respectively), while the number of fetuses reabsorbed by day 30 was higher (75%) for embryos reconstructed with frozen/thawed cells than those reconstructed with cultured cells (31%). A total of 11 kittens from cultured cells and three kittens from frozen/thawed cells were born between days 60 to 64 of gestation. Most kittens died within a few days after birth, although one kitten did survive for 2 months. In Experiment 4, POU5F1 mRNA expression was detected in 25% of blastocysts derived from frozen/thawed cells, whereas 88 and 87% of blastocysts derived from cultured cells and by in vitro fertilization, respectively, expressed POU5F1. We have shown that cell cryopreservation increased the incidence of necrosis and apoptosis and altered epigenetic events in donor cells. Consequently, the number of embryos that cleaved, implanted, and developed to term-gestation and POU5F1 expression in derived blastocysts indirectly was affected.


Gentamicin production by Micromonospora echinospora (Me- 22) in stirred tank reactor: effect of various parameters.
Effect of production medium components, initial starch and soyabean meal concentrations, for the enhanced production of gentamicin by Micromonospora echinospora (Me- 22) was studied in a lab scale stirred tank reactor. Also effect of different aeration (0.5, 1, 2, and 4 vvm) and agitation rates (100, 200, 300 and 400 rpm) in a stirred tank reactor was examined. A maximum gentamicin concentration of 2.68 g l(-1) was achieved in the medium having low concentrations of initial starch (7.5 g l(-1)) and high concentrations of initial soyabean meal (4 g l(-1)). Both aeration and agitation significantly affected gentamicin concentration, productivity and biomass formation. The maximum gentamicin concentration of 4.12 g l(-1) and the highest yield of gentamicin on substrate 0.967 g g(-1) were obtained at impeller speed of 200 rpm and aeration rate of 2 vvm. Under optimal culture conditions in STR the production of gentamicin could be increased 3 fold when compared with shake flask.


Improved Aflatoxins and Fumonisins Forecasting Models for Maize (PREMA and PREFUM), Using Combined Mechanistic and Bayesian Network Modeling-Serbia as a Case Study.
Contamination of maize with aflatoxins and fumonisins is one of the major food safety concerns worldwide. Knowing the contamination in advance can help to reduce food safety risks and related health issues and economic losses. The current study aimed to develop forecasting models for the contamination of maize grown in Serbia with aflatoxins and fumonisins. An integrated modeling approach was used, linking mechanistic modeling with artificial intelligence, in particular Bayesian network (BN) modeling. Two of such combined models, i.e., the prediction model for aflatoxins (PREMA) and for fumonisins (PREFUM) in maize, were developed. Data used for developing PREMA were from 867 maize samples, collected in Serbia during the period from 2012 to 2018, of which 190 were also used for developing PREFUM. Both datasets were split randomly in a model training set and a model validation set. With corresponding geographical and meteorological data, the so-called risk indices for total aflatoxins and total fumonisins were calculated using existing mechanistic models. Subsequently, these risk indices were used as input variables for developing the BN models, together with the longitudes and latitudes of the sites at which the samples were collected and related weather data. PREMA and PREFUM were internally and externally validated, resulting in a prediction accuracy of PREMA of, respectively, 83 and 70%, and of PREFUM of 76% and 80%. The capability of PREMA and PREFUM for predicting aflatoxins and fumonisins contamination using data from the early maize growth stages only was explored as well, and promising results were obtained. The integrated approach combining two different modeling techniques, as developed in the current study, was able to overcome the obstacles of unbalanced data and deficiency of the datasets, which are often seen in historical observational data from the food safety domain. The models provide predictions for mycotoxin contamination at the field level; this information can assist stakeholders of the maize supply chain, including farmers, buyers/collectors, and food safety authorities, to take timely decisions for improved mycotoxin control. The developed models can be further validated by applying them into practice, and they can be extended to other European maize growing areas.


Behavioral Activation, Depression, and Promotion of Health Behaviors: A Scoping Review.
Diverse approaches to the treatment of depressive disorders are necessary to improve evidence-based practice and maximize treatment outcomes. As a result, a range of behaviors and other factors associated with the onset and course of depressive disorders should be examined more comprehensively. Behavioral activation (BA) is a treatment approach to these disorders that can be tailored to address certain health behaviors within the context of depression in an attempt to promote health behaviors whose adoption and maintenance can prove complementary in the treatment of depression. We conducted a scoping review of published studies in which BA-based interventions were used to promote certain health behaviors in individuals with depression. Our search of Medline and the Web of Science identified 336 potential candidate studies. Following screening and with the application of inclusion and exclusion criteria to isolate potentially eligible full-text records, we ultimately identified and evaluated 20 papers that report the nature and efficacy of these modified interventions. Across various domains, including substance use, exercise, medication adherence, and occupational and social success, we found evidence that many-but not all-of the studies we reviewed demonstrated that BA-based interventions were efficacious in promoting the health behavior of interest as well as reducing depressive symptomatology in participants. Implications for more widespread dissemination of such interventions, especially via mobile and web-based platforms due to their accessibility and affordability, are discussed. More research on the feasibility and efficacy of BA-based interventions tailored toward various determinants of health behavior and comorbidities of depressive disorders is warranted.


[Analysis of postsynaptic currents in the phasic and tonic muscle fibers of the extraocular muscles of the rat].
The miniature end-plate currents (MEPCs), miniature postjunctional currents (MPJCs) and mean values of life-time of open ionic channels (tau chan) were compared in experiments on fast and tonic muscle fibres. The rise-time and decay-time constants of MPJCs in tonic fibres were 2.5 and 4-5 times (respectively) longer than those of MEPCs in fast fibres; tau chan in tonic synapses was 2.2 times higher than in fast ones. Inhibition of AChE induced more pronounced prolongation of MEPCs (4.4 times) than MPJCs (1.8 times). As a result, the difference between the time decay of MPJCs and MEPCs reduced to 1.6 times. Thus, a more prolonged decay of MPJCs in tonic fibres in comparison with MEPCs in fast fibres was due to a larger value of tau chan and lower activity of AChE, that favours repetitive interaction between ACh-molecules and choline receptors causing prolongation of the decay of the synaptic response. The time course of MEPCs and tau chan in skeletal slow-twitch muscle fibres was significantly shorter than those in extraocular tonic muscle fibres.


Effect of temperature on hydraulic conductivity of single capillaries.
Paired determinations of the hydraulic conductivity (Lv) of individually perfused capillaries in frog mesentery were measured at 22-25 and 5-8 degrees C. The mean value of the ratio of the hydraulic conductivity at 22-25 to that at 5-8 degrees C was 1.76 +/- 0.22 (SE) in 10 capillaries perfused with albumin in the Ringer perfusate and 1.81 +/- 0.11 in six capillaries perfused with Ringer alone. The ratio of Lp can be accounted for in terms of a 1.60-fold increase in water viscosity between 22-25 and 5-8 degrees C. The results conform to the hypothesis that the structure of the principal pathway for water across the capillary wall is invariant with temperature; models of the water pathway that imply a change in the number of pathways or a change in pathway structure with temperature are incompatible with these results. Temperature gradients across the tissue are negligible in these experiments; calculations show that in the presence of temperature gradients, which occur during tissue cooling or rewarming, transcapillary water flows due to thermoosmosis are likely to occur.


Why is melanoma so metastatic?
Malignant melanoma is one of the most aggressive cancers and can disseminate from a relatively small primary tumor and metastasize to multiple sites, including the lung, liver, brain, bone, and lymph nodes. Elucidating the molecular and genetic changes that take place during the metastatic process has led to a better understanding of why melanoma is so metastatic. Herein, we describe the unique features that distinguish melanoma from other solid tumors and contribute to the malignant phenotype of melanoma cells. For example, although melanoma cells are highly antigenic, they are extremely efficient at evading host immune response. Melanoma cells share numerous cell surface molecules with vascular cells, are highly angiogenic, are mesenchymal in nature, and possess a higher degree of 'stemness' than do other solid tumors. Finally, analysis of melanoma mutations has revealed that the gene expression profile of malignant melanoma is different from that of other cancers. Elucidating these molecular and genetic processes in highly metastatic melanoma can lead to the development of improved treatment and individualized therapy options.


Long-Term Evaluation of Autologous Chondrocyte Implantation: Minimum 7-Year Follow-Up.
The purpose of this study was to report the clinical outcomes of autologous chondrocyte implantation (ACI) procedures performed by a single orthopedic surgeon at a minimum of 7 years follow-up. A retrospective review of prospectively collected data was performed on 29 patients who underwent ACI of the knee between the years of 1998 and 2003. Prospective data were collected to assess changes in standardized outcome measures preoperatively and 2, 4, and 7 years postoperatively. All patients enrolled in the study were also recruited to undergo physical examination when possible. The final cohort consisted of 29 patients with a mean final follow-up time of 8.40 years (range = 7.14-10.88 years). Comparing preoperative scores to 7-year postoperative values, the mean International Knee Documentation Committee (IKDC) score improved from 39.80 to 59.24 (P < 0.001), mean Tegner-Lysholm score increased from 48.07 to 74.17 (P < 0.001), SF-12 physical score improved from 40.38 to 48.66 (P < 0.001), and SF-12 mental score improved from 44.14 to 48.98 (P < 0.05). Significant improvement occurred in Knee Injury and Osteoarthritis Outcome Score (KOOS) pain (56.03 to 80.36), symptoms (54.19 to 74.75), activities of daily living (72.01 to 85.90), sports (23.34 to 55.34), and quality of life (24.56 to 56.03) (P < 0.001). In addition, 7-year postoperative scores were at or near levels seen at 2 years (mean = 2.16; range = 0.94-4.03 years) and 4 years (mean = 4.43; range = 2.16-5.88 years) postoperatively, reflecting durable improvement. Subjectively, on a scale of 1 to 10 (10 being completely satisfied), the mean postoperative satisfaction rate was 8.14. Additionally, 88.9% of the patients would elect to have this surgery again if the same problem was to occur in the contralateral joint. The results of ACI in patients who present with symptomatic, full-thickness chondral defects remain durable at a minimum of 7-year follow-up with persistent, high levels of patient satisfaction. Case series; Level of evidence, IV.


Serum methylmalonic acid by capillary zone electrophoresis using electrokinetic injection and indirect photometric detection.
Elevated methylmalonic acid (MMA) is a superior marker for cobalamin (vitamin B12) deficiency. A rapid assay for serum MMA suitable for clinical specimens has been developed. The specimen is injected electrokinetically after a simple extraction. MMA is measured using indirect photometric detection at 210 nm, based on the displacement of phthalic acid in the running electrolyte; no derivatization is needed. The detection limit is 0.2 mumol/L, well below 0.4 mumol/L, which defines elevated concentrations. The assay is linear up to 100 mumol/L, although specimens above 10 mumol/L are diluted to maximize resolution. The CV is < 10% at 0.30 mumol/L and 0.80 mumol/L. The assay gives results that are comparable to more traditional methods.


Characterization of gold nanoparticle uptake by tomato plants using enzymatic extraction followed by single-particle inductively coupled plasma-mass spectrometry analysis.
Plant uptake and accumulation of nanoparticles (NPs) represent an important pathway for potential human expose to NPs. Consequently, it is imperative to understand the uptake of accumulation of NPs in plant tissues and their unique physical and chemical properties within plant tissues. Current technologies are limited in revealing the unique characteristics of NPs after they enter plant tissues. An enzymatic digestion method, followed by single-particle inductively coupled plasma-mass spectrometry (SP-ICP-MS) analysis, was developed for simultaneous determination of gold NP (AuNP) size, size distribution, particle concentration, and dissolved Au concentration in tomato plant tissues. The experimental results showed that Macerozyme R-10 enzyme was capable of extracting AuNPs from tomato plants without causing dissolution or aggregation of AuNPs. The detection limit for quantification of AuNP size was 20 nm, and the AuNP particle concentration detection limit was 1000 NPs/mL. The particle concentration recoveries of spiked AuNPs were high (79-96%) in quality control samples. The developed SP-ICP-MS method was able to accurately measure AuNP size, size distribution, and particle concentration in the plant matrix. The dosing study indicated that tomato can uptake AuNPs as intact particles without alternating the AuNP properties.


Cell envelope and shape of Escherichia coli: multiple mutants missing the outer membrane lipoprotein and other major outer membrane proteins.
Starting with an Escherichia coli strain missing the outer membrane lipoprotein, multiple mutants were constructed than in addition to this defect miss the outer membrane proteins II, Ia and Ib, or Ia, Ib, and II. In contrast to all single mutants or strains missing the lipoprotein and polypeptides Ia and Ib, drastic influences on the integrity of the outer membrane and cell morphology were observed in mutants without lipoprotein and protein II. Such strains exhibited spherical morphology. They required increased concentrations of electrolytes for optimal growth, and Mg2+ or Ca2+ were the most efficient. These mutants were sensitive to hydrophobic antibiotics and detergents. Electron microscopy revealed abundant blebbing of the outer membrane, and it could clearly be seen that the murein layer was no longer associated with the outer membrane.


[Supporting post of applied medicine].
Whilst specialists don't care much about defining themselves, general practitioners repeatedly are trying to describe their role in modern medicine. Nowadays they prefer to present themselves as gate-keepers at the house of medicine. In that paper it is outlined that this isn't a useful idea.


Carbon nanomaterials in biosensors: should you use nanotubes or graphene?
From diagnosis of life-threatening diseases to detection of biological agents in warfare or terrorist attacks, biosensors are becoming a critical part of modern life. Many recent biosensors have incorporated carbon nanotubes as sensing elements, while a growing body of work has begun to do the same with the emergent nanomaterial graphene, which is effectively an unrolled nanotube. With this widespread use of carbon nanomaterials in biosensors, it is timely to assess how this trend is contributing to the science and applications of biosensors. This Review explores these issues by presenting the latest advances in electrochemical, electrical, and optical biosensors that use carbon nanotubes and graphene, and critically compares the performance of the two carbon allotropes in this application. Ultimately, carbon nanomaterials, although still to meet key challenges in fabrication and handling, have a bright future as biosensors.


Adriamycin in the treatment of resectible and irresectible primary hepatocellular carcinoma.
The treatment of 205 patients with primary hepatocellular carcinoma in Singapore from 1975 to 1980 is reviewed. Adriamycin given at a dose of 40 mg/m(2) intravenously every 4 weeks for irresectible tumours achieved a response rate of 32% alone and has significantly prolonged the median survival time to 4 1/2 months (135 days) in these individuals. Despite this response only 1/34 (3%) patient showed complete remission and is alive at 2 1/2 years. Use of other combinations with Adriamycin e.g. 5 Fluorouracil and Vincristine produced a significant response in the first 16 weeks (p < 0.005) but not thereafter, when completed with the historical group. To improve the response and to consolidate this response, split doses of total hepatic irradiation (to a maximum of 3,150 rads) and alternate day administration of intravenous Adriamycin 10 mg (to a maximum of 90 mg) were given. 12/23 patients (50%) responded. This was followed by a four weekly Adriamycin, 5 F.U. and Vincristine. Median survival time was 5 1/2 months (165 days). Relapse occurred in all patients. Similarly in 5 patients who had resections, early recurrences occurred in 3/5 (60%) within 3 months, inspite of postoperative chemotherapy with Adriamycin, 5 F.U., Vincristine and Cyclophosphamide given every four weeks. In view of the high recurrence rate following effective treatment, further studies on the biological behaviour of these abnormal livers is required.


Critical scattering and incommensurate phase transition in antiferroelectric PbZrO3 under pressure.
Antiferroelectric lead zirconate is the key ingredient in modern ferroelectric and piezoelectric functional solid solutions. By itself it offers opportunities in new-type non-volatile memory and energy storage applications. A highly useful and scientifically puzzling feature of this material is the competition between the ferro- and antiferroelectric phases due to their energetic proximity, which leads to a challenge in understanding of the critical phenomena driving the formation of the antiferroelectric structure. We show that application of hydrostatic pressure drastically changes the character of critical lattice dynamics and enables the soft-mode-driven incommensurate phase transition sequence in lead zirconate. In addition to the long known cubic and antiferroelectric phases we identify the new non-modulated phase serving as a bridge between the cubic and the incommensurate phases. The pressure effect on ferroelectric and incommensurate critical dynamics shows that lead zirconate is not a single-instability-driven system.


The effect of perceived organisational support on burnout among community health nurses in China: the mediating role of professional self-concept.
To examine the mediating effect of professional self-concept on the association between perceived organisational support and burnout among community health nurses in Chengdu, China. Burnout is a common phenomenon among nurses and previous studies have focused on work environmental factors contributing to burnout. Limited studies have examined the effects of perceived organisational support and professional self-concept on burnout among community health nurses. This was a cross-sectional study with 551 community health nurses in Chengdu, China, which included a two-stage sampling method. Structural equation modelling was used to examine the relationships among perceived organisational support, professional self-concept and burnout. The final sample included 456 nurses (82.7%). Perceived organisational support was a significant positive direct predictor for professional self-concept and a significant negative direct predictor for burnout. Professional self-concept was a significant negative direct contributor to burnout. Professional self-concept had a mediating effect on the relationship between perceived organisational support and burnout. Perceived organisational support may result in reduced burnout by facilitating the development of positive professional self-concept. Strategies such as establishing a supportive work environment and professional competence training may be effective methods for burnout prevention and management among community health nurses.


Bespoke Tailoring of Graphenoid Sheets: A Rippled Molecular Carbon Comprising Cyclically Fused Nonbenzenoid Rings.
The incorporation of nonbenzenoid rings into the hexagonal networks of graphenoid nanostructures is of immense importance for electronic, magnetic, and mechanical properties, but the underlying mechanisms of nonbenzenoid ring fusion are rather unexplored. Here, we report the synthesis and characterization of a rippled C84 molecular carbon, which contains 10 nonbenzenoid rings (five-, seven-, and eight-membered rings) that are contiguously fused to give a cyclic geometry. The fused nonbenzenoid rings impart high solubility, configurational stability, multiple reversible redox behaviors, unique aromaticity, and a narrow band gap to the system. Moreover, this carbon nanostructure allows for further functionalization via electrophilic substitution and metalation reactions, enabling access to finely tuned derivatives. Interestingly, both the bowl-shaped and planar conformations of the core in molecular carbon are observed in the solid state. Additionally, this molecular carbon displays ambipolar transport characteristics.


Chest Radiograph and CT Findings in Patients Hospitalized with Breakthrough COVID-19.
To characterize chest radiograph and CT imaging appearance in patients with breakthrough COVID-19 (defined as an illness occurring in patients that previously received a COVID-19 vaccination) in a hospital setting. In this retrospective study, all patients admitted to the hospital between August 26 and September 8, 2021 with a positive SARS-CoV-2 reverse transcription polymerase chain reaction-confirmed infection who were fully vaccinated against COVID-19 were evaluated. Clinical, laboratory data, and outcomes were collected and assessed. All patients had chest imaging performed (either radiography, CT, or a combination of both). Chest radiographs and CTs were assessed and scored on admission and on follow up to determine the extent and type of pulmonary involvement. Descriptive statistics were used. Charts of 60 hospitalized patients that tested positive for SARS-CoV-2 were reviewed for a prior history of COVID-19 vaccination. Eight (13.3%) such patients were identified and included for analysis (mean age, 54 years; range 34-81 years; four women). Patients received either two doses of Pfizer-BioNTech (n = 6), two doses of Moderna (n = 1), or one dose of Johnson and Johnson (n = 1). Five (63%) patients were immunosuppressed at the time of presentation, and six (75%) reported respiratory symptoms. Most of the patients had normal radiographs (4 of 7; 57%). The most common chest CT findings were ground glass opacities (three of five), with mild to moderate severity scores (average, 51; range 8-88). Two patients required intensive care unit admission. However, no patients died and all were either discharged or were on room air without residual respiratory symptoms by the end of the study period. In hospitalized patients with COVID-19 breakthrough illness, normal to mild or moderately positive imaging findings were observed.©RSNA, 2021.


The kinetics of electron transfer between pseudomonas aeruginosa cytochrome c-551 and its oxidase.
The redox reaction between cytochrome c-551 and its oxidase from the respiratory chain of pseudomonas aeruginosa was studied by rapid-mixing techniques at both pH7 and 9.1. The electron transfer in the direction of cytochrome c-551 reduction, starting with the oxidase in the reduced and CO-bound form, is monophasic, and the governing bimolecular rate constants are 1.3(+/- 0.2) x 10(7) M-1 . s-1 at pH 9.1 and 4 (+/- 1) x 10(6) M-1 . s-1 at pH 7.0. In the opposite direction, i.e. mixing the oxidized oxidase with the reduced cytochrome c-551 in the absence of O2, both a lower absorbance change and a more complex kinetic pattern were observed. With oxidized azurin instead of oxidized cytochrome c-551 the oxidation of the c haem in the CO-bound oxidase is also monophasic, and the second-order rate constant is 2 (+/- 0.7) x 10(6) M-1 . s-1 at pH 9.1. The redox potential of the c haem in the oxidase, as obtained from kinetic titrations of the completely oxidized enzyme with reduced azurin as the variable substrate, is 288 mV at pH 7.0 and 255 mV at pH 9.1. This is in contrast with the very high affinity observed in similar titrations performed with both oxidized azurin and oxidized cytochrome c-551 starting from the CO derivative of the reduced oxidase. It is concluded that: (i) azurin and cytochrome c-551 are not equally efficient in vitro as reducing substrates of the oxidase in the respiratory chain of Pseudomonas aeruginosa; (ii) CO ligation to the d1 haem in the oxidase induces a large decrease (at least 80 mV) in the redox potential of the c-haem moiety.


High Availability of the α7-Nicotinic Acetylcholine Receptor in Brains of Individuals with Mild Cognitive Impairment: A Pilot Study Using 18F-ASEM PET.
Emerging evidence supports a hypothesized role for the α7-nicotinic acetylcholine receptor (α7-nAChR) in the pathophysiology of Alzheimer's disease. 18F-ASEM (3-(1,4-diazabicyclo[3.2.2]nonan-4-yl)-6-18F-fluorodibenzo[b,d]thiophene 5,5-dioxide) is a radioligand for estimating the availability of α7-nAChR in the brain in vivo with PET. Methods: In this cross-sectional study, 14 patients with mild cognitive impairment (MCI), a prodromal stage to dementia, and 17 cognitively intact, elderly controls completed 18F-ASEM PET. For each participant, binding in each region of interest was estimated using Logan graphical analysis with a metabolite-corrected arterial input function. Results: Higher 18F-ASEM binding was observed in MCI patients than in controls across all regions, supporting higher availability of α7-nAChR in MCI. 18F-ASEM binding was not associated with verbal memory in this small MCI sample. Conclusion: These data support use of 18F-ASEM PET to examine further the relationship between α7-nAChR availability and MCI.


Effects of dermal exposure to Nicotiana tabacum (Jean Nicot, 1560) leaves in mouse evaluated by multiple methods and tissues.
Tobacco farmers are routinely exposed to complex mixtures of the compounds present in tobacco leaves, including organic and inorganic pesticides. Penetration through skin is the most significant route of uptake in occupational exposure to chemicals, including dust and liquids containing toxic and carcinogenic substances. This study evaluates the genotoxic effect of tobacco leaves with and without dermal exposure to flumetralin in Mus musculus, determining cell damage by the micronucleus test and the Comet assay as well as antioxidant enzyme activities and hematologic parameters. Nicotine was used as positive control. Blood samples were collected for 0, 3, 24 and 48 h exposure periods, and DNA damage by Comet assay and micronucleus test was evaluated for all these periods. Bone marrow and liver cells were also evaluated for the 48 h exposure period. Significant differences between Comet assay results in blood cells from animals exposed to tobacco leaves with and without pesticide were found in 24 and 48 h exposure periods in relation to negative control. Bone marrow cells from the group exposed to leaves with pesticide (48 h) also demonstrated significant increase in DNA damage. Concerning the micronucleus test, only animals exposed to tobacco leaves without pesticide (24 h) showed increase in frequency of micronuclei when compared to the negative control. Oxidative stress activities also were demonstrated for different groups. The results demonstrate the injury effect caused by tobacco leaves in different Mus musculus tissues, suggesting that the effects of dermal exposure to tobacco leaves are caused by complex mixtures present in the plant, but mainly by nicotine.


Increase in non-professional phagocytosis during the progression of cell cycle.
Homotypic or heterotypic internalization of another, either living or necrotic cell is currently in the center of research interest. The active invasion of a living cell called entosis and cannibalism of cells by rapidly proliferating cancers are prominent examples. Additionally, normal healthy tissue cells are capable of non-professional phagocytosis. This project studied the relationship between non-professional phagocytosis, individual proliferation and cell cycle progression. Three mesenchymal and two epithelial normal tissue cell lines were studied for homotypic non-professional phagocytosis. Homotypic dead cells were co-incubated with adherent growing living cell layers. Living cells were synchronized by mitotic shake-off as well as Aphidicolin-treatment and phagocytotic activity was analyzed by immunostaining. Cell cycle phases were evaluated by flow cytometry. Mesenchymal and epithelial normal tissue cells were capable of internalizing dead cells. Epithelial cells had much higher non-professional phagocytotic rates than mesenchymal cells. Cells throughout the entire cell cycle were able to phagocytose. The phagocytotic rate significantly increased with progressing cell cycle phases. Mitotic cells regularly phagocytosed dead cells, this was verified by Nocodazole and Colcemid treatment. Taken together, our findings indicate the ability of human tissue cells to phagocytose necrotic neighboring cells in confluent cell layers. The origin of the cell line influences the rate of cell-in-cell structure formation. The higher cell-in-cell structure rates during cell cycle progression might be influenced by cytoskeletal reorganization during this period or indicate an evolutionary anchorage of the process. Recycling of nutrients during cell growth might also be an explanation.


Management of the Rectal Stump after Subtotal Colectomy Operations for Inflammatory Bowel Disease in the Era of Immunologic Therapy: A Two-Centre Cohort Study.
Inflammatory bowel disease (IBD) often requires surgical resection, such as subtotal colectomy operations to alleviate symptoms. However, IBD also has an inherently increased risk of colorectal dysplasia and cancer. Despite the well-accepted surveillance guidelines for IBD patients with an intact colon, contemporaneous decision-making models on rectal stump surveillance is sparse. This study looks at the fate of rectal stumps in IBD patients following subtotal colectomy. This is a two-centre retrospective observational cohort study. Patients were identified from NHS Grampian and NHS Highland surgical IBD databases. Patients that had subtotal colectomy between January 01, 2010 and December 31, 2017 were included with the follow-up end date on April 1, 2021. Socio-demographics, diagnosis, medical and surgical management data were collected from electronic records. Of 250 patients who had subtotal colectomy procedures, only one developed a cancer in their rectal stump (0.4%) over a median follow-up of 80 months. A higher than expected 72% of patients had ongoing symptoms from their rectal stumps. Surveillance was varied and inconsistent. However, no surveillance, flexible sigmoidoscopy, or MRI identified dysplastic or neoplastic disease. Based on our results, we estimate that the prevalence of rectal cancer is lower than previously reported. Surveillance strategy of rectal stump varied as no current guidelines exist and hence is an important area for future study. Given the relatively low frequency of rectal cancer in these patients, and the low level of evidence available in this field, we would propose a registry-based approach to answering this important clinical question.


Physical activity and mental health in a student population.
A growing body of literature indicates that physical activity can have beneficial effects on mental health. However, previous research has mainly focussed on clinical populations, and little is known about the psychological effects of physical activity in those without clinically defined disorders. The present study investigates the association between physical activity and mental health in an undergraduate university population based in the United Kingdom. One hundred students completed questionnaires measuring their levels of anxiety and depression using the Hospital Anxiety and Depression Scale (HADS) and their physical activity regime using the Physical Activity Questionnaire (PAQ). Significant differences were observed between the low, medium and high exercise groups on the mental health scales, indicating better mental health for those who engage in more exercise. Engagement in physical activity can be an important contributory factor in the mental health of undergraduate students.


Relationships between the histological grade of cutaneous mast cell tumours in dogs, their survival and the efficacy of surgical resection.
The histological grade of 340 cutaneous mast cell tumours derived from 280 dogs was determined by an established histological grading system; 87 of the tumours (26 per cent) were well differentiated, 199 (59 per cent) were intermediately differentiated and 54 (16 per cent) were poorly differentiated. The one-year survival rates for the dogs with tumours of these three grades were significantly different (P = 0.0001), being 100 per cent, 92 per cent and 46 per cent, respectively. The median survival time for the dogs with poorly differentiated tumours was 278 days, significantly shorter than that for the dogs with either intermediately or well-differentiated tumours, which were both over 1300 days. Regrowth of the tumours was identified in 10 (19 per cent) of the dogs with poorly differentiated tumours, 12 (6 per cent) of the dogs with intermediately differentiated tumours and one of the dogs with well-differentiated tumours; only three of the tumours which regrew had initially had complete margins. The results suggest that wide surgical margins are not a prerequisite for a successful long-term outcome in dogs with well-differentiated cutaneous mast cell tumours.


Concentration of 3H-8-methoxypsoralen and its metabolites in the rat lens and eye after a single oral administration.
Concentrations of 3H-8-methoxypsoralen (MOP), its lipid- and water-soluble metabolites, tritiated water, and an insoluble compound have been measured in rat serum, lens, and eye tissue by liquid scintillation, thin-layer chromatography, and other techniques. The radioactivity has been measured in albino and pigmented rats up to 1 week after medication, and 3H-8-MOP and metabolite concentrations have been measured in pigmented rats from 10 min to 24 hr after oral administration by stomach tube of a solution of 3H-8-MOP (1 mg/kg body weight). The animals were kept in the dark. The maximum 3H-8-MOP concentrations were seen 10 min after medication in all the organs examined. The concentrations (in microgram/kg) at 10 min, 2 hr, and 24 hr, respectively, after medication were: serum, 698, 55, and 0.8; lens, 11.3, 5.2, and 1.4; other eye tissue, 393, 23, and 2.5. Water soluble metabolites might accumulate in the lens, but the absolute concentration is small. There was no accumulation of metabolites in the other organs. The considerable amount of radioactivity still present in tissues 1 week after a single medication was a result of the formation of tritiated water from the degraded 3H-8-MOP.


Differences in the protein composition of bovine retinal rod outer segment disk and plasma membranes isolated by a ricin-gold-dextran density perturbation method.
The plasma membrane and disk membranes of bovine retinal rod outer segments (ROS) have been purified by a novel density-gradient perturbation method for analysis of their protein compositions. Purified ROS were treated with neuraminidase to expose galactose residues on plasma membrane-specific glycoproteins and labeled with ricin-gold-dextran particles. After the ROS were lysed in hypotonic buffer, the plasma membrane was dissociated from the disks by either mild trypsin digestion or prolonged exposure to low ionic strength buffer. The dense ricin-gold-dextran-labeled plasma membrane was separated from disks by sucrose gradient centrifugation. Electron microscopy was used to follow this fractionation procedure. The dense red pellet primarily consisted of inverted plasma membrane vesicles containing gold particles; the membrane fraction of density 1.13 g/cc consisted of unlabeled intact disks and vesicles. Ricin-binding studies indicated that the plasma membrane from trypsin-treated ROS was purified between 10-15-fold. The protein composition of plasma membranes and disks was significantly different as analyzed by SDS gels and Western blots labeled with lectins and monoclonal antibodies. ROS plasma membrane exhibited three major proteins of 36 (rhodopsin), 38, and 52 kD, three ricin-binding glycoproteins of 230, 160, and 110 kD, and numerous minor proteins in the range of 14-270 kD. In disk membranes rhodopsin appeared as the only major protein. A 220-kD concanavalin A-binding glycoprotein and peripherin, a rim-specific protein, were also present along with minor proteins of 43 and 57-63 kD. Radioimmune assays indicated that the ROS plasma membrane contained about half as much rhodopsin as disk membranes.


Coordinate regulation of phosphoenolpyruvate carboxylase and phosphoenolpyruvate carboxykinase by light and CO2 during C4 photosynthesis.
The aim of this study was to investigate the relationship between the phosphorylation and activation states of phosphoenolpyruvate carboxykinase (PEPCK) and to investigate how the phosphorylation states of PEPCK and phosphoenolpyruvate carboxylase (PEPC) are coordinated in response to light intensity and CO(2) concentration during photosynthesis in leaves of the C(4) plant Guinea grass (Panicum maximum). There was a linear, reciprocal relationship between the phosphorylation state of PEPCK and its activation state, determined in a selective assay that distinguishes phosphorylated from nonphosphorylated forms of the enzyme. At high photon flux density and high CO(2) (750 microL L(-1)), PEPC was maximally phosphorylated and PEPCK maximally dephosphorylated within 1 h of illumination. The phosphorylation state of both enzymes did not saturate until high light intensities (about 1,400 micromol quanta m(-2) s(-1)) were reached. After illumination at lower light intensities and CO(2) concentrations, the overall change in phosphorylation state was smaller and it took longer for the change in phosphorylation state to occur. Phosphorylation states of PEPC and PEPCK showed a strikingly similar, but inverse, pattern in relation to changes in light and CO(2). The protein phosphatase inhibitor, okadaic acid, promoted the phosphorylation of both enzymes. The protein synthesis inhibitor, cycloheximide, blocked dark phosphorylation of PEPCK. The data show that PEPC and PEPCK phosphorylation states are closely coordinated in vivo, despite being located in the mesophyll and bundle sheath cells, respectively.


Interrelationships between yeast ribosomal protein assembly events and transient ribosome biogenesis factors interactions in early pre-ribosomes.
Early steps of eukaryotic ribosome biogenesis require a large set of ribosome biogenesis factors which transiently interact with nascent rRNA precursors (pre-rRNA). Most likely, concomitant with that initial contacts between ribosomal proteins (r-proteins) and ribosome precursors (pre-ribosomes) are established which are converted into robust interactions between pre-rRNA and r-proteins during the course of ribosome maturation. Here we analysed the interrelationship between r-protein assembly events and the transient interactions of ribosome biogenesis factors with early pre-ribosomal intermediates termed 90S pre-ribosomes or small ribosomal subunit (SSU) processome in yeast cells. We observed that components of the SSU processome UTP-A and UTP-B sub-modules were recruited to early pre-ribosomes independently of all tested r-proteins. On the other hand, groups of SSU processome components were identified whose association with early pre-ribosomes was affected by specific r-protein assembly events in the head-platform interface of the SSU. One of these components, Noc4p, appeared to be itself required for robust incorporation of r-proteins into the SSU head domain. Altogether, the data reveal an emerging network of specific interrelationships between local r-protein assembly events and the functional interactions of SSU processome components with early pre-ribosomes. They point towards some of these components being transient primary pre-rRNA in vivo binders and towards a role for others in coordinating the assembly of major SSU domains.


Dual roles for glucokinase in glucose homeostasis as determined by liver and pancreatic beta cell-specific gene knock-outs using Cre recombinase.
Glucokinase (GK) gene mutations cause diabetes mellitus in both humans and mouse models, but the pathophysiological basis is only partially defined. We have used cre-loxP technology in combination with gene targeting to perform global, beta cell-, and hepatocyte-specific gene knock-outs of this enzyme in mice. Gene targeting was used to create a triple-loxed gk allele, which was converted by partial or total Cre-mediated recombination to a conditional allele lacking neomycin resistance, or to a null allele, respectively. beta cell- and hepatocyte-specific expression of Cre was achieved using transgenes that contain either insulin or albumin promoter/enhancer sequences. By intercrossing the transgenic mice that express Cre in a cell-specific manner with mice containing a conditional gk allele, we obtained animals with either a beta cell or hepatocyte-specific knock-out of GK. Animals either globally deficient in GK, or lacking GK just in beta cells, die within a few days of birth from severe diabetes. Mice that are heterozygous null for GK, either globally or just in the beta cell, survive but are moderately hyperglycemic. Mice that lack GK only in the liver are only mildly hyperglycemic but display pronounced defects in both glycogen synthesis and glucose turnover rates during a hyperglycemic clamp. Interestingly, hepatic GK knock-out mice also have impaired insulin secretion in response to glucose. These studies indicate that deficiencies in both beta cell and hepatic GK contribute to the hyperglycemia of MODY-2.


[Plasmid DNA of Salmonella derby].
S. derby K 89 isolated from sick children have been found to contain plasmids which determine the resistance of these organisms to penicillin and chloramphenicol. 3 types of plasmid DNA with molecular weights of 40. 12 and 8 megadaltons (Md) have been detected by electrophoretic analysis. The dissociation of the large plasmid (40 Md) into 3 independent replicons has been observed. The circular, covalently closed forms of the DNA cointegrate and its constituent replicons have been identified by electron microscopy. In S. derby K 89 grown in broth prepared from dried meat concentrate the loss of these plasmids is observed, and the culture thus becomes sensitive to penicillin and chloramphenicol. This suggests that dried meat concentrate contains some factor capable of eliminating the plasmids.


Coronary endothelial function is preserved with chronic endothelin receptor antagonism in experimental hypercholesterolemia in vitro.
Hypercholesterolemia is associated with increased circulating and tissue endothelin-1 immunoreactivity, decreased nitric oxide (NO) activity, and altered endothelial function. We tested the hypothesis that chronic endothelin receptor antagonism preserves endothelial function and increases NO in experimental porcine hypercholesterolemia. Pigs were randomized to 3 groups: Group 1, a 2% high-cholesterol (HC) diet alone (n=7); group 2, RO-48-5695, a combined endothelin receptor antagonist, and an HC diet (n=8); and group 3, ABT-627, a selective endothelin-A receptor antagonist, and an HC diet (n=8). Coronary epicardial and arteriolar endothelial function was determined by a dose-response relaxation to bradykinin (10(-11) to 10(-6) mol/L), in all groups and in pigs maintained on a normal diet. Plasma total oxidized products of NO (NO(x)) were determined by chemiluminescence at baseline and after 12 weeks. Bradykinin-stimulated coronary epicardial and arteriolar relaxation in group 1 was attenuated compared with normal-diet controls. This relaxation was normalized with endothelin receptor antagonism. Plasma NO(x) decreased after 12 weeks in group 1 (-74.8+/-5.5%). This decrease was attenuated in the endothelin receptor antagonist groups (group 2, -28.2+/-15.0%; group 3, -38.9+/-20.6%). Chronic endothelin receptor antagonism preserves coronary endothelial function and increases NO in hypercholesterolemia. This study supports a role of endothelin-1 in the regulation of NO activity and suggests a possible therapeutic role for endothelin receptor antagonists in hypercholesterolemia.