A magnetosome chain viewed as a bio-elastic magnet.
In light of the coarse-grained Monte Carlo numerical simulation method, the magnetosome chain stability of magnetotactic bacteria is analysed and discussed. This discrete chain of magnetic nanoparticles, encapsulated in a lipid membrane and flanked by filaments, orients bacteria in the geomagnetic field as a compass needle. Each magnetosome is a magnetite or greigite nanocrystal encapsulated in a soft lipid shell. This structure is modelled by a hard core with a magnetic dipole embedded and a cloud of electric dipoles which are able to move and rotate over the magnetic spherical core. In the present paper, some of the many possibilities of the model by varying the control parameters of the system are explored. Magnetic particles arrange in long linear clusters when the coating is removed. However, linear but twisted chains of magnetic particles emerge when there are electric dipoles in the coating shell. A unique linear and straight chain is not observed in any 3D numerical simulation; this result is in agreement with a real living system of bacteria in a geomagnetic field when proteins that form the filament are absent. Finally, the stability and magnetization of a magnetosome chain of 30 beads in one dimension set up are discussed resembling a real chain. The results suggest that a magnetosome chain not only orients bacteria but also should be considered as a potential storage of elastic energy.


The changes of trace elements, malondialdehyde levels and superoxide dismutase activities in pregnancy with or without preeclampsia.
Increased free radical activity and lipid peroxidation may be implicated in the pathogenesis of preeclampsia. This study was initiated to assess antioxidant enzyme and trace metals's status in preeclampsia. The comparison was made between the pregnant women with or without preeclampsia and healthy controls in this study. Samples were obtained from 24 normal nonpregnant (controls), 30 normal pregnant and 21 preeclamptic women in the third trimester. Lipid peroxidation end product, malondialdehyde (MDA), free radical scavenging enzyme activity, superoxide dismutase (SOD) and serum zinc (Zn), copper (Cu) levels were measured in either plasma/serum or erythrocytes of patients. Data were analyzed statistically using Student's t-test. In the preeclamptic group malondialdehyde, Cu levels were significantly increased, while Zn and SOD levels were significantly decreased compared to normal control group and healthy pregnant women. Our findings give support that radical scavenging SOD is consumed by the increased lipid peroxidation in preeclampsia. This data may indicate an involvement of free radicals in the pathophysiology of preeclampsia. This study suggests a relationship between increased MDA, Cu levels and decreased SOD, Zn levels in pregnancy and preeclampsia.


Inverse CeO2/CuO catalyst as an alternative to classical direct configurations for preferential oxidation of CO in hydrogen-rich stream.
A novel inverse CeO(2)/CuO catalyst for preferential oxidation of CO in H(2)-rich stream (CO-PROX) has been developed on the basis of a hypothesis extracted from previous work of the group (JACS 2007, 129, 12064). Possible separation of the two competing oxidation reactions involved in the process (of CO and H(2), respectively) is the key to modulation of overall CO-PROX activity and is based on involvement of different sites as most active ones for each of the two reactions. Achievement of large size CuO particles and adequate CeO(2)-CuO interfacial configurations in the inverse catalyst apparently allows appreciable enhancement of the catalytic properties of this kind of system for CO-PROX, constituting an interesting alternative to classic direct configurations so far explored for this process. Reasons for such behavior are analyzed on the basis of operando-XRD, -XAFS, and -DRIFTS studies.


Marek's disease in turkeys. II. Characterization of the viral glycoprotein B gene and antigen of a turkey strain of Marek's disease virus.
Marek's disease virus (MDV) causes immunosuppression and tumors in chickens. As sporadic cases of Marek's disease (MD) were recorded in turkeys, the antigenic and genomic characteristics of the MDV glycoprotein B (gB) gene and antigen of turkeys were compared to the chicken MDV gB. The whole chicken and turkey gB genes were sequenced and found identical. By immunoblotting of infected-cell culture lysates using chicken convalescent and gB monoclonal antibodies, the antigenic epitopes of the chicken and turkey viruses were found to differ. The turkey MDV had a unique epitope, compared to the chicken MDV and compared with our previous findings. While the chicken MDV had two epitope types, heat-labile but dithiothreitol (DTT)-stable and heat-stable but DTT-labile, the turkey MDV gB epitope is both heat and DTT-labile.


Assessing consistency in clinical trials with two subgroups and binary endpoints: A new test within the logistic regression model.
In late stage drug development, the experimental drug is tested in a diverse study population within the relevant indication. In order to receive marketing authorization, robust evidence for the therapeutic efficacy is crucial requiring investigation of treatment effects in well-defined subgroups. Conventionally, consistency analyses in subgroups have been performed by means of interaction tests. However, the interaction test can only reject the null hypothesis of equivalence and not confirm consistency. Simulation studies suggest that the interaction test has low power but can also be oversensitive depending on sample size-leading in combination with the actually ill-posed null hypothesis to findings regardless of clinical relevance. In order to overcome these disadvantages in the setup of binary endpoints, we propose to use a consistency test based on the interval inclusion principle, which is able to reject heterogeneity and confirm consistency of subgroup-specific treatment effects while controlling the type I error. This homogeneity test is based upon the deviation between overall treatment effect and subgroup-specific effects on the odds ratio scale and is compared with an equivalence test based on the ratio of both subgroup-specific effects. Performance of these consistency tests is assessed in a simulation study. In addition, the consistency tests are outlined for the relative risk regression. The proposed homogeneity test reaches sufficient power in realistic scenarios with small interactions. As expected, power decreases for unbalanced subgroups, lower sample sizes, and narrower margins. Severe interactions are covered by the null hypothesis and are more likely to be rejected the stronger they are.


LPA signaling is required for dopaminergic neuron development and is reduced through low expression of the LPA1 receptor in a 6-OHDA lesion model of Parkinson's disease.
Lysophosphatidic acid (LPA) is a bioactive phospholipid that activates at least five known G-protein-coupled receptors (GPCRs): LPA1-LPA5. The nervous system is a major locus for LPA1 expression. LPA has been shown to regulate neuronal proliferation, migration, and differentiation during central nervous system development as well as neuronal survival. Furthermore, deficient LPA signaling has been implicated in several neurological disorders including neuropathic pain and schizophrenia. Parkinson's disease (PD) is a neurodegenerative movement disorder that results from the loss of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNc). The specific molecular pathways that lead to DA neuron degeneration, however, are poorly understood. The influence of LPA in the differentiation of mesenchymal stem cells (MSCs) into DA neurons in vitro and LPA1 expression in a 6-hydroxydopamine (6-OHDA) lesion model of PD in vivo were examined in the present study. LPA induced neuronal differentiation in 80.2 % of the MSC population. These MSCs developed characteristic neuronal morphology and expressed the neuronal marker, neuron-specific enolase (NSE), while expression of the glial marker, glial fibrillary acidic protein (GFAP), was absent. Moreover, 27.6 % of differentiated MSCs were positive for tyrosine hydroxylase (TH), a marker for DA neurons. In the 6-OHDA PD rat model, LPA1 expression in the substantia nigra was significantly reduced compared to control. These results suggest LPA signaling via activation of LPA1 may be necessary for DA neuron development and survival. Furthermore, reduced LPA/LPA1 signaling may be involved in DA neuron degeneration thus contributing to the pathogenesis of PD.


Comparative study of the antioxidant activity of the essential oils of five plants against the H2O2 induced stress in Saccharomyces cerevisiae.
The purpose of this work was to investigate the protective effect of five essential oils (EOs); Rosmarinus officinalis, Thymus vulgaris, Origanum compactum Benth., Eucalyptus globulus Labill. and Ocimum basilicum L.; against oxidative stress induced by hydrogen peroxide in Saccharomyces cerevisiae. The chemical composition of the EOs was analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS). The in vitro antioxidant activity was evaluated and the protective effect of EOs was investigated. Yeast cells were pretreated with different concentrations of EOs (6.25-25 µg/ml) for an hour then incubated with H2O2 (2 mM) for an additional hour. Cell viability, antioxidants (Catalase, Superoxide dismutase and Glutathione reductase) and metabolic (Succinate dehydrogenase) enzymes, as well as the level of lipid peroxidation (LPO) and protein carbonyl content (PCO) were evaluated. The chemical composition of EOs has shown the difference qualitatively and quantitatively. Indeed, O. compactum mainly contained Carvacrol, O. basilicum was mainly composed of Linalool, T. vulgaris was rich in thymol, R. officinalis had high α-Pinene amount and for E. globulus, eucalyptol was the major compound. The EOs of basil, oregano and thyme were found to possess the highest amount of total phenolic compounds. Moreover, they have shown the best protective effect on yeast cells against oxidative stress induced by H2O2. In addition, in a dose dependent manner of EOs in yeast medium, treated cells had lower levels of LPO, lower antioxidant and metabolic enzymes activity than cells exposed to H2O2 only. The cell viability was also improved. It seems that the studied EOs are efficient natural antioxidants, which can be exploited to protect against damages and serious diseases related to oxidative stress.


Effects of pH, sample size, and solvent partitioning on recovery of soluble phenolic acids and isoflavonoids in leaves and stems of red clover (Trifolium pratense cv. Kenland).
Several extraction parameters were tested to determine optimal conditions for extracting phenolics from leaves and stems of red clover (Trifolium pratense L. cv. Kenland), with the goal of using extracts in bioassays and in assessment of phenolic profiles. HPLC-UV profiles were compared before and after partitioning a methanolic extract of soluble phenolics with ethyl acetate-ethyl ether (1:1, v/v). The effect of extract pH on the partitioning of phenolics into the ethyl acetate-ethyl ether (EtOAc-Et2O) phase was evaluated, and several tissue weights were extracted to determine a minimum amount that could be extracted without loss of information. HPLC profiles of soluble phenolics were similar in the methanolic extracts and the partitioned EtOAc-Et2O extracts. However, recoveries in unpartitioned extracts were 2- to 4-fold greater than in the acidified, partitioned extracts. Also, recovery was considerably affected by the pH to which extracts were adjusted prior to partitioning. In extracts acidified to pH 2, recoveries were 2- to 7-fold higher than in extracts partitioned at pH 6. In extracts prepared from 250, 120, or 60 mg of tissue, peak areas of methanolic extracts were directly proportional to the amount of tissue extracted.


Gliadin peptide P31-43 localises to endocytic vesicles and interferes with their maturation.
Celiac Disease (CD) is both a frequent disease (1:100) and an interesting model of a disease induced by food. It consists in an immunogenic reaction to wheat gluten and glutenins that has been found to arise in a specific genetic background; however, this reaction is still only partially understood. Activation of innate immunity by gliadin peptides is an important component of the early events of the disease. In particular the so-called "toxic" A-gliadin peptide P31-43 induces several pleiotropic effects including Epidermal Growth Factor Receptor (EGFR)-dependent actin remodelling and proliferation in cultured cell lines and in enterocytes from CD patients. These effects are mediated by delayed EGFR degradation and prolonged EGFR activation in endocytic vesicles. In the present study we investigated the effects of gliadin peptides on the trafficking and maturation of endocytic vesicles. Both P31-43 and the control P57-68 peptide labelled with fluorochromes were found to enter CaCo-2 cells and interact with the endocytic compartment in pulse and chase, time-lapse, experiments. P31-43 was localised to vesicles carrying early endocytic markers at time points when P57-68-carrying vesicles mature into late endosomes. In time-lapse experiments the trafficking of P31-43-labelled vesicles was delayed, regardless of the cargo they were carrying. Furthermore in celiac enterocytes, from cultured duodenal biopsies, P31-43 trafficking is delayed in early endocytic vesicles. A sequence similarity search revealed that P31-43 is strikingly similar to Hrs, a key molecule regulating endocytic maturation. A-gliadin peptide P31-43 interfered with Hrs correct localisation to early endosomes as revealed by western blot and immunofluorescence microscopy. P31-43 and P57-68 enter cells by endocytosis. Only P31-43 localises at the endocytic membranes and delays vesicle trafficking by interfering with Hrs-mediated maturation to late endosomes in cells and intestinal biopsies. Consequently, in P31-43-treated cells, Receptor Tyrosine Kinase (RTK) activation is extended. This finding may explain the role played by gliadin peptides in inducing proliferation and other effects in enterocytes from CD biopsies.


A Qualitative Study Exploring Models of Supportive Care in Men and Their Partners/Caregivers Affected by Metastatic Prostate Cancer.
To explore the experiences of patients with metastatic prostate cancer and their partners/caregivers, as well as an interprofessional team, with a nurse-led multimodality supportive care intervention. . Qualitative study. . National Health Service (NHS), Tayside, Scotland. . 19 patients, 7 partners/caregivers, and 7 interprofessional members from four hospitals in NHS, Tayside, Scotland. . 33 semistructured interviews were conducted to explore patients' and partners/caregivers' experiences of supportive care, and a framework approach was used to analyze the data. . Men and their partners/caregivers experienced a range of unmet physical, psychological, and informational supportive care needs. The participants in the intervention group reported overall high satisfaction with the use of holistic needs assessments and self-management plans, with a decrease in unmet needs compared to the standard of care over time. The prostate cancer specialist nurse was perceived as the hub of survivorship care. Members of the interprofessional team perceived benefit in the nurse-led multimodal supportive care intervention. . An emphasis needs to be placed on personalizing care, with supportive care interventions targeted to individual needs. . Care can be improved by incorporating holistic needs assessment to target specialized interventions for optimized, individualized care plans. An intervention seminar encouraged self-management and self-efficiency, leading to greater satisfaction for participants.


Effect of short-term fasting on plasma concentrations of leptin and other hormones and metabolites in dairy cattle.
We determined the effects of short-term fasting and refeeding on temporal changes in plasma concentrations of leptin, insulin, insulin-like growth factor- 1 (IGF-1), growth hormone (GH), glucose, and nonesterified fatty acids (NEFA), in early lactating cows, non-lactating pregnant cows, and postpubertal heifers. In experiment 1, Holstein cows in early lactation were either fed ad libitum (Control, n=5) or feed deprived for 48 h (Fasted, n=6). Plasma leptin, insulin, and glucose concentrations rapidly declined (P<0.05) within 6h, and IGF-1 by 12h, but all these variables sharply returned to control levels (P>0.10) within 2h of refeeding. Plasma NEFA and GH concentrations were elevated (P<0.05) by 4 and 36 h of fasting and returned to control levels (P>0.10) by 8 and 24h after refeeding, respectively. In experiment 2, four ruminally cannulated pregnant non-lactating Holstein cows were used in a cross-over design and were fasted for 48 h (Fasted) or fasted with partial evacuation of rumen contents (Fasted-Evac). The plasma variables measured did not differ (P>0.10) between Fasted and Fasted-Evac cows. Plasma leptin, insulin, and IGF-1 concentrations were reduced by 10, 6, and 24h of fasting, respectively, in Fasted-Evac cows; and these variables were reduced by 24h in Fasted cows (P<0.05). Plasma glucose levels were reduced (P<0.05) by 48 h of fasting in both groups of fasted animals. Plasma NEFA and GH levels were increased (P<0.05) by 12 and 48 h of fasting, respectively. In experiment 3, postpubertal Holstein heifers were either fed ad libitum (Control, n=4) or feed deprived for 72 h (Fasted, n=5). Concentrations of leptin, insulin, IGF-1, and glucose in plasma were reduced (P<0.05) by 24, 10, 24, and 48 h of fasting, respectively. Plasma NEFA concentrations increased (P<0.05) by 4h, of fasting while GH levels were not significantly (P>0.10) affected by fasting. Collectively, our data provide evidence that plasma leptin concentrations are reduced with short-term fasting and rebound on refeeding in dairy cattle with the response dependent on the physiological state of the animals. Compared to the rapid induction of hypoleptinemia with fasting of early lactation cows, the fasting-induced hypoleptinemia was delayed in non-lactating cows and postpubertal heifers.


[A case of advanced breast cancer with multiple bone metastases responding to docetaxel and high-dose toremifene as fourth-line chemo-endocrine therapy].
A 55-year old woman, who underwent left mastectomy (Bt+Ax), was revealed to have sternum metastasis by postoperative 99mTc bone scanning(T1bN1M1). She received daily aromatase inhibitor (anastrozole), as a primary systemic endocrine therapy, and biweekly pamidronate for metastatic breast cancer. However, she depended on folk medicine a year later, at which time the primary treatment was discontinued. Another year later, the bone metastases developed with increased serum levels of tumor markers (CEA, CA19-9, and NCC-ST-439). Then, she underwent three different regimens of systemic chemo-endocrine therapy over the following three years, including CAF+MPA as the first-line, paclitaxel (PTX) + anastrozole as the second-line, and S-1+anastrozole as the third-line regimen. She recently completed 10 courses of the fourth-line regimen[tri-weekly docetaxel (DOC) and high-dose toremifene (TOR 120 mg/day)], which reduced levels of 99mTc accumulation in the multiple bone metastases and levels of the serum tumor markers to the normal range. No severe adverse events occurred except peripheral thrombovasculitis (grade 2) in her left anterior arm during the fourth regimen. She recently maintains the current status by taking a regular dose (40 mg/day) of toremifene for 5 months. Combination treatment with DOC and high-dose TOR can be one of the worthwhile regimens as systemic chemo-endocrine therapy for patients with advanced breast cancer who develop bone metastases.


Androgens, adrenal androgen precursors, and their metabolism in untreated primary tumors and lymph node metastases of human prostatic cancer.
Activities of several steroid metabolizing enzymes (steroid sulfate-sulfatase, 17 beta-hydroxysteroid dehydrogenase, 5 alpha-reductase, and 3 alpha beta-hydroxysteroid dehydrogenase) as well as total tissue content and subcellular distribution (nuclear-extranuclear) of several androgen precursors, active androgens, and androgen deactivation products (DHEA sulfate, DHEA, 5-androstenediol, 4-androstenedione, testosterone, DHT, and 3 alpha-androstanediol) were quantified in primary tumors and lymph node metastases of human prostatic cancer obtained from patients without previous endocrine manipulation. Primary tumors were compared to benign parts of the same prostates, and the metastases were compared to their primary tumors. All enzymes and steroids found in benign prostatic tissues could also be detected in the malignant tissues. Even the capacity to accumulate active androgens in the nuclei was found to be unchanged in nearly all of the samples. Lower activities of hormone-dependent enzymes were observed in the cancers, suggesting a less efficient utilization of hormonal stimuli. Most striking changes found in the malignant tissues were a subtotal loss of 5 alpha-reductase activity and a metabolic shift to testosterone, which was more pronounced in samples from metastatic disease as compared to samples from non-metastatic disease. In conclusion, primary tumors and metastases of prostatic cancers not treated by endocrine manipulations retain their androgen receptor system and possess the same capacity to metabolize adrenal androgen precursors along the pathway to DHT as benign prostatic tissue. Consequently, they should be able to use at least androstenedione for production of active androgens directly in the target tissue.


Effects of added chelated trace minerals, organic selenium, yeast culture, direct-fed microbials, and Yucca schidigera extract in horses. Part I: Blood nutrient concentration and digestibility.
The objective of this study was to test the hypothesis that feed additives such as chelated minerals, organic Se, yeast culture, direct-fed microbials, and Yucca schidigera extract would improve nutrient digestibility when included in an equine diet. Horses (Quarter Horse geldings 4.5 to 16 yr of age; mean BW 522 kg ± 46 kg) were acclimated to 100% pelleted diets formulated with (ADD) and without (CTRL) commercially available sources of the aforementioned additives followed by a 14-d collection period of feces and urine. Chelated sources of Cu, Zn, Mn and Co were utilized versus sulfated forms, at a 100% replacement rate. No significant differences among apparent the digestibility of DM, ADF, or NDF (P= 0.665, P = 0.866, P = 0.747, respectively) were detected between dietary treatments. Likewise, no differences in apparent digestibility of Cu (P = 0.724), Zn (P = 0.256), Mn (P = 0.888), Co (P = 0.71), or Se (P = 0.588) were observed. No differences were observed in serum Cu, Mn, or Co concentrations between ADD and CTRL at acclimation or collection time points (P > 0.05). While no difference in serum Zn concentrations were observed between ADD and CTRL groups at acclimation (P > 0.05), they were statistically higher at the collection time period for horses consuming CTRL (P < 0.0001). Whole blood Se concentration was greater in the CTRL group versus the ADD group both at acclimation (P = 0.041) and collection (P = 0.005) time periods. In reference to time, serum Cu concentrations increased (P = 0.012) for animals consuming CTRL, but not ADD (P > 0.05). Serum Zn concentrations of horses consuming both ADD (P = 0.021) and CTRL (P < 0.0001) increased over time from acclimation to collection time points. No time differences (P > 0.05) were observed in serum Mn concentrations. Serum Co concentrations increased over time in horses consuming both ADD (P = 0.001) and CTRL (P = 0.021). From acclimation to collection, whole blood Se concentration increased for horses consuming CTRL (P = 0.01) but not for ADD (P > 0.05). The results of this study indicate no effect on nutrient digestibility due to the inclusion of chelated minerals, organic Se, yeast culture, direct-fed microbials, and Yucca schidigera extract for horses at maintenance.


[Adverse events causing discontinuation of donepezil for Alzheimer's dementia].
The objective was to investigate the frequency of adverse events (AE) as a cause for discontinuation of donepezil treatment for Alzheimer's dementia (DAT) in a geriatric memory unit. Five-year retrospective study of 123 donepezil-treated patients diagnosed with DAT or mixed dementia in a geriatric memory unit. The material covers all patients treated with donepezil and surveyed for 12 months in the memory unit during the period from 14th March 2001 to 7th April 2006. Among the 123 patients, 106 (86%) suffered from DAT and 17 (14%) suffered from mixed dementia. A total of 100 (81%) were female while 23 (19%) were male. The median age was 84 years. In all, 26 (21%) patients discontinued treatment due to AE. The most frequent AE were nausea/vomiting, diarrhoea and loss of appetite. In 21% of the cases treatment was discontinued within 12 months due to AE. The most frequent AEs were nausea/vomiting, diarrhoea and loss of appetite. For the most part discontinuation took place within the initial three months. However, some treatments were not discontinued until after six months; consequently, treatment should be supervised and patients should have easy access to a memory unit in the case of AE.


[A case of renal infarction treated by thrombolytic therapy with systemic intravenous injection of tPA (tissue plasminogen activator)].
A 55-year-old woman affected by mitralism presented with severe right flank pain of sudden onset. Biochemical examinations showed elevated serum lactate dehydrogenase, and abdominal enhanced computed tomography (CT) demonstrated hypoperfusion of the right kidney. Infarction of the right kidney was highly suspected, and she was immediately treated by systemic intravenous injection of 12,000,000 units of tissue plasminogen activator (tPA) per day for 3 days and 120,000 units of urokinase per day for 8 days. After the thrombolytic therapy, abdominal enhanced CT revealed marked improvement of enhancement of right renal parenchyma and decrease of serum LDH. Although thrombolytic therapy with selective intraarterial infusion is considered to be a useful treatment modality for renal infarction, systemic administration of tPA may also be effective judging from the clinical course of the present case.


Greater leftward lateralization of the inferior frontal gyrus in second language learners with higher syntactic abilities.
There is a great individual variability for acquiring syntactic knowledge in a second language (L2). Little is, however, known if there is any anatomical basis in the brain for individual differences in syntactic acquisition. Here we examined brain structures in 95 nonnative speakers of English, including 78 high-school students and 17 adult international students. We found a significant correlation between the performance of a syntactic task and leftward lateralization of a single region in the triangular part (F3t) of the inferior frontal gyrus, which has been proposed as the grammar center. Moreover, this correlation was independent of the performance of a spelling task, age, gender, and handedness. This striking result suggests that the neural basis for syntactic abilities in L2 is independent of that for lexical knowledge in L2, further indicating that the individual differences in syntactic acquisition are related to the lateralization of the grammar center.


Oral mucosa equivalents, prevascularization approaches, and potential applications.
Oral mucosa equivalents (OMEs) have been used as in vitro models (eg, for studies of human oral mucosa biology and pathology, toxicological and pharmacological tests of oral care products), and clinically to treat oral defects. However, the human oral mucosa is a highly vascularized tissue and implantation of large OMEs can fail due to a lack of vascularization. To develop equivalents that better resemble the human oral mucosa and increase the success of implantation to repair large-sized defects, efforts have been made to prevascularize these constructs. The aim of this narrative review is to provide an overview of the human oral mucosa structure, common approaches for its reconstruction, and the development of OMEs, their prevascularization, and in vitro and clinical potential applications. Articles on non-prevascularized and prevascularized OMEs were included, since the development and applications of non-prevascularized OMEs are a foundation for the design, fabrication, and optimization of prevascularized OMEs. Several studies have reported the development and in vitro and clinical applications of OMEs and only a few were found on prevascularized OMEs using different approaches of fabrication and incorporation of endothelial cells, indicating a lack of standardized protocols to obtain these equivalents. However, these studies have shown the feasibility of prevascularizing OMEs and their implantation in animal models resulted in enhanced integration and healing. Vascularization in tissue equivalents is still a challenge, and optimization of cell culture conditions, biomaterials, and fabrication techniques along with clinical studies is required.


Moderated Mediation Model from Stress to Burnout among Health Professionals.
Objectives: It is unclear whether poor health is a contributing factor or a consequence of burn-out. We aimed to explore the relationship among job stress, physical health, mental health, burnout, and coping strategies among health professionals using a moderated mediation model. Methods: Health professionals in Taiwan were invited using a stratified and probability proportional to size sampling. A structure equation model (SEM) was used to examine relationships among job stress, burnout, and physical/mental health. The bootstrapping approach for moderated mediation analysis was then used to explore the role of coping strategies. Results: Totally 935 health professionals participated. The SEM model revealed that job stress was inversely associated with physical and mental health. The direct association between job stress and burn-out was insignificant. Moderated mediation analysis revealed that job stress influenced burnout through physical health and mental health, and that the coping strategy of support-seeking moderated the indirect effect only through mental health. Conclusions: Support-seeking is a good coping strategy for health professionals to buffer the harmful effects of job stress on mental health.


Dirofilaria immitis in a child from the Russian Federation.
An immature female worm, Dirofilaria immitis, was isolated from the scrotum of a 14-month-old child. This is the first identification of human dirofilariosis caused by D. immitis in a relatively Northern region (Moscow) of the Russian Federation. The parasite was diagnosed by means of morphological examination of the worm, confirmed by PCR assay. This case raises questions about the range of the D. immitis distribution among humans in Russia. To better understand the geographical distribution of dirofilarioses, detailed clinical and epidemiological information should be collected from human cases with appropriate laboratory confirmation.


Issues concerning androgen replacement therapy in postmenopausal women.
A decrease in blood androgen levels is well documented in women who experience natural or surgical menopause. This change may be associated with various negative effects on bone metabolism in addition to psychosocial and sexuality aspects of life. A review of published information on androgen replacement therapy shows that major benefits may be achieved; unfortunately, only minimal quality information is available to help clinicians make decisions about this type of therapy. In this review, we point out the potential benefits and risks to bone, lipid and carbohydrate metabolism, and sexuality; in addition, we discuss potential risks of neoplasms and virilizing somatic changes. Long-term, physiologic, and well-designed androgen replacement studies should be performed to obtain the knowledge needed to guide therapy in this important area.


Castleman Disease of the Thorax: Clinical, Radiologic, and Pathologic Correlation: From the Radiologic Pathology Archives.
Castleman disease is a complex lymphoproliferative disease pathologically divided into two subtypes, the hyaline vascular variant (HVV) and the plasma cell variant (PCV). The HVV is the most common, is thought to represent a benign neoplasm of lymph node stromal cells, and is treated with surgical resection. It is most commonly found in the mediastinum, where it classically appears as a unicentric, avidly enhancing mass at computed tomography (CT) and magnetic resonance imaging. This appearance can mimic other avidly enhancing mediastinal masses, and location, clinical history, laboratory data, and nuclear medicine single photon emission CT (SPECT) and positron emission tomography (PET) studies can help narrow the differential diagnosis. Multicentric Castleman disease (MCD), which in the majority of cases is composed of the PCV, is an aggressive lymphoproliferative disorder associated with human herpesvirus infection, interleukin 6 dysregulation, and other systemic disorders. While it can be difficult to differentiate MCD from lymphoma, the presence of avidly enhancing lymph nodes can suggest the diagnosis. The purpose of this article is to review the clinical, immunologic, and pathologic findings associated with both unicentric Castleman disease and MCD and discuss how the imaging findings correlate with the pathophysiology of the disease.


SMARCA4 biology in alveolar rhabdomyosarcoma.
Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children and phenocopies a muscle precursor that fails to undergo terminal differentiation. The alveolar subtype (ARMS) has the poorest prognosis and represents the greatest unmet medical need for RMS. Emerging evidence supports the role of epigenetic dysregulation in RMS. Here we show that SMARCA4/BRG1, an ATP-dependent chromatin remodeling enzyme of the SWI/SNF complex, is prominently expressed in primary tumors from ARMS patients and cell cultures. Our validation studies for a CRISPR screen of 400 epigenetic targets identified SMARCA4 as a unique factor for long-term (but not short-term) tumor cell survival in ARMS. A SMARCA4/SMARCA2 protein degrader (ACBI-1) demonstrated similar long-term tumor cell dependence in vitro and in vivo. These results credential SMARCA4 as a tumor cell dependency factor and a therapeutic target in ARMS.


RNAi-directed downregulation of vacuolar H(+) -ATPase subunit a results in enhanced stomatal aperture and density in rice.
Stomatal movement plays a key role in plant development and response to drought and salt stress by regulating gas exchange and water loss. A number of genes have been demonstrated to be involved in the regulation of this process. Using inverse genetics approach, we characterized the function of a rice (Oryza sativa L.) vacuolar H(+)-ATPase subunit A (OsVHA-A) gene in stomatal conductance regulation and physiological response to salt and osmotic stress. OsVHA-A was constitutively expressed in different rice tissues, and the fusion protein of GFP-OsVHA-A was exclusively targeted to tonoplast when transiently expressed in the onion epidermal cells. Heterologous expression of OsVHA-A was able to rescue the yeast mutant vma1Δ (lacking subunit A activity) phenotype, suggesting that it partially restores the activity of V-ATPase. Meanwhile, RNAi-directed knockdown of OsVHA-A led to a reduction of vacuolar H(+)-ATPase activity and an enhancement of plasma membrane H(+)-ATPase activity, thereby increasing the concentrations of extracellular H(+) and intracellular K(+) and Na(+) under stress conditions. Knockdown of OsVHA-A also resulted in the upregulation of PAM3 (plasma membrane H(+)-ATPase 3) and downregulation of CAM1 (calmodulin 1), CAM3 (calmodulin 3) and YDA1 (YODA, a MAPKK gene). Altered level of the ion concentration and the gene expression by knockdown of OsVHA-A probably resulted in expanded aperture of stomatal pores and increased stomatal density. In addition, OsVHA-A RNAi plants displayed significant growth inhibition under salt and osmotic stress conditions. Taken together, our results suggest that OsVHA-A takes part in regulating stomatal density and opening via interfering with pH value and ionic equilibrium in guard cells and thereby affects the growth of rice plants.


Exploration of the nicotinamide-binding site of the tankyrases, identifying 3-arylisoquinolin-1-ones as potent and selective inhibitors in vitro.
Tankyrases-1 and -2 (TNKS-1 and TNKS-2) have three cellular roles which make them important targets in cancer. Using NAD(+) as a substrate, they poly(ADP-ribosyl)ate TRF1 (regulating lengths of telomeres), NuMA (facilitating mitosis) and axin (in wnt/β-catenin signalling). Using molecular modelling and the structure of the weak inhibitor 5-aminoiso quinolin-1-one, 3-aryl-5-substituted-isoquinolin-1-ones were designed as inhibitors to explore the structure-activity relationships (SARs) for binding and to define the shape of a hydrophobic cavity in the active site. 5-Amino-3-arylisoquinolinones were synthesised by Suzuki-Miyaura coupling of arylboronic acids to 3-bromo-1-methoxy-5-nitro-isoquinoline, reduction and O-demethylation. 3-Aryl-5-methylisoquinolin-1-ones, 3-aryl-5-fluoroisoquinolin-1-ones and 3-aryl-5-methoxyisoquinolin-1-ones were accessed by deprotonation of 3-substituted-N,N,2-trimethylbenzamides and quench with an appropriate benzonitrile. SAR around the isoquinolinone core showed that aryl was required at the 3-position, optimally with a para-substituent. Small meta-substituents were tolerated but groups in the ortho-positions reduced or abolished activity. This was not due to lack of coplanarity of the rings, as shown by the potency of 4,5-dimethyl-3-phenylisoquinolin-1-one. Methyl and methoxy were optimal at the 5-position. SAR was rationalised by modelling and by crystal structures of examples with TNKS-2. The 3-aryl unit was located in a large hydrophobic cavity and the para-substituents projected into a tunnel leading to the exterior. Potency against TNKS-1 paralleled potency against TNKS-2. Most inhibitors were highly selective for TNKSs over PARP-1 and PARP-2. A range of highly potent and selective inhibitors is now available for cellular studies.


Enrofloxacin/florfenicol loaded cyclodextrin metal-organic-framework for drug delivery and controlled release.
We presented an antibiotic-loaded γ-cyclodextrin metal-organic framework that delivered antibiotics suitable for the treatment of bacterial infections. The γ-cyclodextrin metal-organic framework was developed using γ-cyclodextrin and potassium ion via the ultrasonic method. The antibiotic (florfenicol and enrofloxacin) was primarily encapsulated into the pore structures of γ-CD-MOF, which allowed the sustained release of antibiotics over an extended period of time in vitro and in vivo. Notably, antibiotics-loaded γ-CD-MOF showed much superior activity against bacteria than free antibiotics (lower MIC value) and displayed better long-lasting activity (longer antibacterial time). The antibiotics-loaded γ-CD-MOF showed nontoxic and perfect biocompatibility to mammalian cells and tissues both in vitro and in vivo. These materials thus represent a novel drug-delivery device suitable for antibiotic therapy. This research is of great significance for reducing the generation of bacterial resistance and providing new ideas for the application of γ-CD-MOF.


Contribution of complement component C3 and complement receptor type 3 to carbohydrate-dependent uptake of oligomannose-coated liposomes by peritoneal macrophages.
Peritoneal macrophages (PEMs) preferentially and rapidly take up oligomannose-coated liposomes (OMLs) and subsequently mature to induce a Th-1 immune response following administration of OMLs into the peritoneal cavity. Here, we examine the contributions of complement component C3 and complement receptor type 3 (CR3) to carbohydrate-dependent uptake of OMLs by PEMs. Effective uptake of OMLs into PEMs in vitro was observed only in the presence of peritoneal fluid (PF), and OMLs incubated with PF were incorporated by PEMs in vitro in the absence of PF. These phenomena were inhibited by methyl-alpha-mannoside, N-acetylglucosamine or EDTA, but not by galactose. Pull-down analysis followed by peptide mass fingerprinting of PF-treated OMLs indicated that the OMLs were opsonized with complement fragment iC3b. In vivo uptake of OMLs by PEMs was inhibited by intraperitoneal injection of an antibody against CR3, a receptor for iC3b, and OML uptake by PEMs in the peritoneal cavity was not observed in C3-deficient mice. Thus, our results indicate that OMLs are opsonized with iC3b in a mannose-dependent manner in the peritoneal cavity and then incorporated into PEMs via CR3.


Validation of the activPAL activity monitor in children with hemiplegic gait patterns resultant from cerebral palsy.
Current investigation of treatment outcomes by clinicians is currently hampered by a lack of clinically viable tools. The use of activity monitors specifically validated for a population could help resolve this situation. The purpose of the study was to validate an activity monitor for monitoring children with cerebral palsy. The study was designed to validate the duration of time spent sitting, standing and walking and the number of steps taken when being measured by the activPAL activity monitor. A validation study was undertaken. Observations of participants were carried out while completing a specifically designed activity course using video footage, which were then compared to the output from the activity monitor. The activity monitor was found to be valid for the time spent standing and walking, the number of steps taken and the number of transitions. The results demonstrated that the monitor may prove useful to clinicians as a measurement outcome device for children with hemiplegic gait patterns resultant from cerebral palsy. However, the sensitivity of the device is variable, and further investigations are necessary to confirm it would also be able to detect minor changes after interventions. The monitor may provide clinicians with a simplistic tool that is easily utilised, to enable audit exercises of current and future treatments.


Global inhibition of Lef1/Tcf-dependent Wnt signaling at its nuclear end point abrogates development in transgenic Xenopus embryos.
Analysis of canonical Wnt signaling during vertebrate development by means of knock-out or transgenic approaches is often hampered by functional redundancy as well as pathway bifurcations downstream of the manipulated components. We report the design of an optimized chimera capable of blocking transcriptional activation of Lef1/Tcf-beta-catenin target genes, thus enabling intervention with the canonical Wnt pathway at its nuclear end point. This construct was made hormone-inducible, both functionally and transcriptionally, and was transgenically integrated in Xenopus embryos. Down-regulation of target genes was clearly observed upon treatment of these embryos with dexamethasone. In addition, exposure of variously aged transgenic embryos to dexamethasone caused complex phenotypes with many new but also several recognizable features stemming from inhibition of canonical Wnt signaling. At least in some tissues, a significant reduction in cell proliferation and an increase in programmed cell death appeared to underlie these phenotypes. Our inducible transgenic system can serve a broad range of experimental settings designed to unveil new functional aspects of Lef1/Tcf-beta-catenin signaling during vertebrate embryogenesis.


Evaluation of air-Q™ intubating laryngeal airway as a conduit for tracheal intubation in infants--a pilot study.
Air-Q™ intubating laryngeal airway (ILA) has been used successfully as a conduit for tracheal intubation in pediatric difficult airway. However, its use as an airway device and conduit for intubation in infants is not yet evaluated. The primary objective was to evaluate ILA as a conduit for tracheal intubation in infants, and secondary objectives were to evaluate ILA in terms of ease of insertion and ventilation, oropharyngeal leak pressure (OLP), glottic view, and complications. Twenty infants with normal airway were included. After induction of general anesthesia and neuromuscular blockade, ILA (size 1.0/1.5) was inserted. Fiberoptic bronchoscope (FOB)-guided intubation and removal of ILA with the stabilizing stylet was attempted. The mean age and weight of the infants were 7.5 ± 2.3 months and 7.3 ± 1.8 kg. The ILA sizes 1.0 and 1.5 were inserted in 10 infants each according to the weight of the infants. The mean OLP was 18.5 ± 1.8 cm H₂O, and mean time for insertion of ILA was 13.3 ± 3.9 s. Glottic view was grade I in 6, grade II in 1, grade III in 4, and grade IV in nine infants. Tracheal intubation was successful in 19/20 infants. The mean endotracheal tube (ETT) insertion time and mean total time (ILA insertion to the confirmation of ETT placement) were 95.6 ± 32.3 s and 306.42 ± 120.2 s respectively. The ILA is a safe and easy-to-use supraglottic airway device for intubation with FOB in infants with normal airway. Insertion and removal of ILA are easy with satisfactory oropharyngeal seal and ventilation.


Calcium concentrations in dental plaque of patients with juvenile and adult periodontitis.
Calcium concentrations in plaque of patients with juvenile (JP) and adult (AP) periodontitis were compared. These calcium concentrations in JP patients were significantly (P less than 0.01) lower than in AP patients. Because JP patients are not intense calculus-formers, the finding that they have low calcium concentrations in plaque agrees with the authors' previous suggestion that calculus formation is a marker of high calcium concentrations in plaque. JP patients have good caries resistance in general, as was the case in this study. The finding that they have low calcium concentrations in plaque conflicts with the view that caries-resistant dentitions are usually associated with high concentrations of calcium in plaque.


KRDS, a peptide derived from human lactotransferrin, inhibits thrombin-induced thromboxane synthesis by a cyclooxygenase-independent mechanism.
KRDS, a tetrapeptide from human lactotransferrin, inhibits thrombin-induced platelet aggregation, secretion and thromboxane (TX) synthesis without interfering with phospholipase C (PLC) beta activation, since in previous work we have shown that Ca2+ mobilization and phosphorylation of the myosin light chain kinase (20 kDa) and pleckstrin (47 kDa) were normal. However, the inhibition of arachidonic acid-induced aggregation in the presence of KRDS is accompanied by normal TX synthesis suggesting that it does not interfere with the cyclooxygenase activity. To elucidate further the mechanisms of action of this peptide we tested its effect on U46619-induced platelet activation. KRDS inhibits U46619-induced platelet aggregation time- and dose-dependently without inhibiting the phosphorylation of pleckstrin. This suggests that the PLC pathway is not affected and that the inhibitory effect of KRDS is not due to and uncoupling of TXA2 from its receptor. In addition to the PLC pathway, protein tyrosine kinases play a major role in platelet signal transduction mechanisms. At least 7 tyrosine-phosphorylated proteins are detected upon stimulation of platelets by thrombin. KRDS strongly inhibits the tyrosine-phosphorylated substrates, in particular two 100-105 kDa substrates which are related to GP IIb/IIIa activation and platelet aggregation. The absence of TX synthesis observed in the presence of KRDS could be due to the inactivation of cPLA2 since the latter needs tyrosine phosphorylation to be activated, thus explaining the inhibitory action of KRDS on platelet functions.


Efficacy and Safety of Topical Therapy With Botanical Products for Melasma: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.
Botanical products have been increasingly popular in topical therapies for melasma, as presumed safer and milder than fully synthetic products. Although the efficacy of different topical botanicals has recently been substantiated through randomized controlled trials (RCTs), there is a lack of sufficiently pooled evidence on their efficacy and safety for the treatment of melasma. Herein, a systematic review and meta-analysis was conducted on the efficacy and safety of topical botanical products for the treatment of melasma, following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). All RCTs on the use of topical botanical products for the treatment of melasma in humans were included, except for trials enrolling pregnant patients. The primary outcome was Melasma Area and Severity Index (MASI) or its variation. The secondary outcomes included Mexameter® reading, melasma improvement evaluated by participants, and any reported adverse events (AEs). As a result, twelve eligible trials comprising 695 patients with melasma from 6 different countries were included. The topical botanical products contained active ingredients which varied among trials as follows: herb-derived molecule, extracts of a single herb, and extracts of compound herbs. Topical therapy with botanical products significantly improved melasma with a large effect on MASI reduction (SMD -0.79, 95% CI -1.14 to -0.44, p < 0.00001), and a moderate effect on Mexameter® reading reduction (SMD -0.52, 95% CI -0.81 to 0.23, p = 0.0005), when compared with placebo. It also showed a similar improvement of melasma with a better safety profile (RR 0.37, 95% CI 0.15-0.88, p = 0.02), when compared with active-comparators. Botanical products were well-tolerated across studies, with no serious AEs reported. Despite the limitations such as small sample size, short duration of follow up and varied botanical products, this work still represents the best level of evidence currently available on topical use of botanical products on melasma. Moreover, it should be noted that more well-designed studies are needed before recommending topical botanical products as a viable treatment option for melasma. PROSPERO, identifier: CRD42021256328.


[A case of acute autonomic, sensory and motor neuropathy (AASMN)--less favorable response of the autonomic dysfunctions to IVIg treatment].
We report a rare case of acute autonomic, sensory and motor neuropathy (AASMN). The patient, a 26-year-old woman, developed fever and common cold around January 20, 2001 and was admitted because of abdominal pain due to ileus on January 30. After admission, the patient complained of muscle weakness and numbness in the extremities, difficulty in seeing with the right eye, and dysuria. Neurologically, marked orthostatic hypotension, right tonic pupil, distal dominant moderate muscle weakness in extremities, areflexia in both lower limbs, glove and stocking type of paresthesia, and neurogenic atonic bladder were noted. Sensation to pin prick, light touch, temperature, and vibration were markedly impaired in upper limbs and below the level of the 5th thoracic cord. Cerebrospinal fluid examination revealed albumino-cytologic dissociation. Peripheral nerve conduction study revealed lower limb dominant axonal type impairment of sensory conduction and slight impairment of motor conduction velocity. Clinical autonomic testings revealed dysfunction of both sympathetic and parasympathetic systems. As having AASMN, she was given the intravenous high-dose immunoglobulin (IVIg) therapy twice. After IVIg, the sensory and motor symptoms improved remarkably, but pandysautonomia did not. To our knowledge, this is the first report of AASMN treated by IVIg, and the notable clinical feature in this case was the favorable motor and sensory recovery to IVIg, as opposed to poor autonomic outcome.


Ratiometric electrogenerated chemiluminescence sensor based on a designed anti-fouling peptide for the detection of carcinoembryonic antigen.
In this paper, an effective and accurate ratiometric electrochemiluminescence (ECL) system based on Au-luminol and CdS quantum dots (CdS QDs) as signal probes was constructed for detecting carcinoembryonic antigen (CEA). Polyaniline (PANI) and gold nanoparticles (AuNPs) strongly enhanced the electronic transfer efficiency and the specific area of the modified sensing surface, and improved the detection sensitivity. CdS QDs functionalized DNA strands functioned as cathode ECL emitters, and the aptamer of CEA modified with Au-luminol and quencher cyanine dye (Cy5) fluorophore functioned as anode ECL emitters. After the DNA capture probe combined with CEA aptamer, the negative potential ECL response of the CdS QDs was quenched because of electrochemiluminescence resonance energy transfer (ERET) between the Cy5 fluorophore and CdS QDs. However, the positive potential ECL response of Au-luminol can still be detected. The negative potential ECL response of CdS QDs was recovered, and the positive potential ECL response of Au-luminol decreased after CEA combined with its aptamer to take Cy5 and Au-luminol off the sensing interface. Additionally, the peptide provided effective anti-fouling performance in the detection of complex samples. The ratiometric ECL sensor with anti-fouling ability can sensitively determine the concentration of CEA in human serum.


Structural and functional recovery of microbial biofilms after a decrease in copper exposure: influence of the presence of pristine communities.
The present study aimed at assessing the recovery of phototrophic and heterotrophic biofilm communities after a decrease in copper exposure. An original experiment was designed to evaluate the possible influence of non-exposed (i.e. pristine) communities (e.g. via immigration processes) in recovery dynamics. Laboratory channels were used to study the structural and functional changes in microbial communities after a 4-week Cu exposure period in the presence and absence of pristine biofilms. When pristine biofilms were present, phototrophic communities recovered within 6 weeks, both in terms of biomass, structure and photosynthetic activity. Recovery processes were also detected using the PICT approach. In contrast, in the absence of pristine communities, all of the Cu-induced changes recorded in the phototrophic communities remained throughout the recovery period. Regardless of the presence or absence of pristine biofilms, the decrease in Cu exposure did not abolish Cu-induced changes in bacterial community structure, whereas functional recovery (based on beta-glucosidase activity) was complete in both recovery contexts. These results revealed that microbial community response to a decrease in Cu exposure differs between phototrophic and heterotrophic communities. The presence of pristine communities greatly influences the structural and functional recovery of phototrophic communities, suggesting an important role of microbial immigration processes, but have far less influence on the recovery trajectory of heterotrophic communities.


The public health impact of Lyme disease in Maryland.
The purpose of this study was to estimate the health burden imposed by Lyme disease (LD) in Maryland during 1992 and 1993. A cross-sectional 1-in-15 survey of physicians (total, 1200) in Maryland was conducted to estimate the incidence of diagnosed LD, presumptive cases of LD, patients with tick bites, and diagnostic tests ordered for LD. Results show that LD is underreported by 10- to 12-fold in Maryland, that 80% of cases are managed by primary care physicians, and that there is discordance between the actual clinical treatment of patients and the recommended approach. In addition, the much greater numbers of patients treated for presumptive LD, seen and given prophylaxis for tick bites, and having diagnostic tests indicate that real and perceived LD is a far greater public health problem and uses more medical resources than official surveillance data suggest.


Transesterification in Vitrimer Polymers Using Bifunctional Catalysts: Modeled with Solution-Phase Experimental Rates and Theoretical Analysis of Efficiency and Mechanisms.
Recently, thermoset vitrimer polymers have shown significant promise for structural applications because of their ability to be reshaped and remolded due to their covalent adaptive network (CAN). In these vitrimers, the transesterification reaction is responsible for the CAN, where the efficiency of the reaction is controlled either by organic or by organometallic catalysts. Understanding the mechanism of the transesterification reaction in the bulk phase using direct experimental techniques is extremely difficult due to the highly cross-linked complex structure of thermosetting vitrimers. Therefore, we use solution-phase experiments to investigate the catalytic efficiency and to guide density functional theory (DFT) simulations of the transesterification reaction mechanism with catalysts triazabicyclodecene (TBD), zinc acetate (Zn(OAc)2), 1-methylimidazole (1-MI), and dibutyltin oxide (DBTO). The estimated catalytic efficiency from the detailed DFT reaction path calculations follows the order TBD ≳ DBTO ≳ Zn(OAc)2 > 1-MI, which agrees with the experimental results. In addition to reaction path modeling, the mechanism and the relative rates of the transesterification reaction are analyzed with the assistance of Fukui indices as a measure of electrophilicity and nucleophilicity of atomic sites and with partial charges. It was found that the sum of the nucleophilicity index of the base and the electrophilicity index of the acid of the bifunctional catalysts correlates with the SN2 transition state and tetrahedral intermediate energies, which are related to the barrier of the rate-limiting step. This correlation provides a hypothesis for computational prescreening of potentially better catalysts that have an index in a range of values. These results provide a basis for understanding an important part of the mechanism of transesterification in vitrimer systems and may assist with designing new catalysts.


The smallest resonance energy transfer acceptor for tryptophan.
The utility of diazirine ligands as acceptors in resonance energy transfer (RET) distance measurements with tryptophan or tryptophan analogues as donor is reported. The principle is demonstrated for a diazirine derivative of d-mannitol, 2-azi-2-deoxy-d-arabino-hexitol, and single-tryptophan-containing mutants of the mannitol transporter, EIImtl, from E. coli. The Förster distance is 10 A for the tryptophan-diazirine donor-acceptor couple, allowing the measurement of distances up to 17 A. The versatility of tryptophan as an intrinsic spectroscopic probe in protein chemistry and the small size of the diazirine group makes this a very attractive donor-acceptor couple for accurate RET distance information in protein chemistry.


Light-Activated Nanodevice for On-Demand Imaging of miRNA in Living Cells via Logic Assembly.
Low-abundance biomarker amplification detection systems have been widely used to detect miRNAs; however, "always active" systems are insufficient for high spatial and temporal control of miRNAs. Here, we constructed a light-activated nanodevice (LAN) based on DNA nanotechnology for high spatial and temporal precision detection of low-abundance miRNA. Light-activated hairpin probes and triple-helix molecular switches were modified on the surface of gold nanoparticles (AuNPs) to trigger miRNA on-demand imaging analysis by UV light activation. In the presence of both UV light and miRNA, the LAN releases hairpin DNA and completes the hybridization chain reaction (HCR) with the conformation-altered triple-helix molecular switch, enabling fluorescence imaging of low-abundance miRNAs in living cells. The current work provides an opportunity to develop light-activated signal amplification sensors that can accurately image miRNAs on-demand in both temporal and spatial dimensions.


[Revolution in diagnostic microbiology. Needs, driving forces and consequences].
Introducing molecular biological technology into medical microbiology promotes deeper insight into the epidemiology, pathogenesis, diagnostics and treatment of infectious diseases. New technology has helped to highlight the complicated biological interaction between host and microbes and has created a need for more advanced technology both in microbiological diagnostics and in microbiological science. Extra-professional factors like the high prestige of new techniques have also been of considerable significance. Medical microbiologists will become an important link between the inventors of new technology and the physicians who diagnose and treat the patients. Through these contacts and their diagnostic laboratory work, they will become important generators and communicators of knowledge. In this connection it will be important to represent reason in introducing technology in medical microbiological diagnostics. Since patients are expected to play a more active part in health care, communicating knowledge to patients and the population in general will become an obligation.


Outcomes of myocardial fibrosis in patients undergoing transcatheter aortic valve replacement.
We sought to investigate the relevance of myocardial fibrosis, assessed by mid-wall fibrosis risk (MFR) score, with respect to left ventricular (LV) reverse remodelling following transcatheter aortic valve replacement (TAVR). Between January 2010 and March 2015, we enrolled 207 patients in whom baseline MFR, which includes age, sex, high-sensitivity cardiac troponin I, presence of strain pattern on electrocardiography, and peak aortic valve velocity, as well as one-year follow-up echocardiography was available. LV reverse remodelling was defined as a >10% reduction in LV end-diastolic volume index (LVEDVi). A higher MFR score (≥52) was associated with increased LVEDVi and with decreased LV ejection fraction as well as higher baseline NT-proBNP levels (p<0.05 for all). One year after the TAVR procedure, a higher MFR score was associated with a decreased probability of LV reverse remodelling (OR 0.33, 95% CI: 0.23-0.87; p=0.03), which was independent of baseline echocardiographic parameters and comorbidities. In contrast, there was no significant difference in five-year mortality between patients with lower and higher MFR scores (57.9% vs 60.5%, p=0.66). A higher MFR score is associated with reduced LV reverse remodelling at one-year follow-up, whereas the MFR score does not appear to correlate with long-term mortality after TAVR.


A Review on the Design of Hydrogels With Different Stiffness and Their Effects on Tissue Repair.
Tissue repair after trauma and infection has always been a difficult problem in regenerative medicine. Hydrogels have become one of the most important scaffolds for tissue engineering due to their biocompatibility, biodegradability and water solubility. Especially, the stiffness of hydrogels is a key factor, which influence the morphology of mesenchymal stem cells (MSCs) and their differentiation. The researches on this point are meaningful to the field of tissue engineering. Herein, this review focus on the design of hydrogels with different stiffness and their effects on the behavior of MSCs. In addition, the effect of hydrogel stiffness on the phenotype of macrophages is introduced, and then the relationship between the phenotype changes of macrophages on inflammatory response and tissue repair is discussed. Finally, the future application of hydrogels with a certain stiffness in regenerative medicine and tissue engineering has been prospected.


Stereotactic radiosurgery. VII. Radiosurgery versus conventionally-fractionated radiotherapy in the treatment of cavernous sinus meningiomas.
Twenty-eight patients with meningiomas involving the cavernous sinus were referred to the radiosurgical service at St. Bartholomew's Hospital 1989-1998. The majority (8/13) of patients with small (< 3 cm diameter) tumours received radiosurgery whereas the majority of large tumours (> 3 cm diameter; 12/15) received conventionally-fractionated radiotherapy. Other treatment recommendations were fractionated radiosurgery (one case) and conventionally-fractionated radiotherapy to the whole meningeal base and a radiosurgery boost (three cases). Fractionated radiosurgery is optimal where the target volume is small, but abuts critically sensitive nervous system. There have been no progressions of disease at relatively early follow-up. It is argued that subtotal excision followed by appropriate dose radiation therapy is often a strong competitor to attempted radical excision given the attendant morbidity of this latter operation for meningiomas at this site. Patients referred for radiation therapy are best served by a department with both radiosurgery and conventional radiation therapy facilities.


Pulmonary artery catheterization in acute myocardial infarction complicated by cardiogenic shock: A review of contemporary literature.
Acute myocardial infarction (AMI) with left ventricular (LV) dysfunction patients, the most common cause of cardiogenic shock (CS), have acutely deteriorating hemodynamic status. The frequent use of vasopressor and inotropic pharmacologic interventions along with mechanical circulatory support (MCS) in these patients necessitates invasive hemodynamic monitoring. After the pivotal Evaluation Study of Congestive Heart Failure and Pulmonary Artery Catheterization Effectiveness trial failed to show a significant improvement in clinical outcomes in shock patients managed with a pulmonary artery catheter (PAC), the use of PAC has become less popular in clinical practice. In this review, we summarize currently available literature to summarize the indications, clinical relevance, and recommendations for use of PAC in the setting of AMI-CS.


Shock-Driven Decomposition of Polymers and Polymeric Foams.
Polymers and foams are pervasive in everyday life, as well as in specialized contexts such as space exploration, industry, and defense. They are frequently subject to shock loading in the latter cases, and will chemically decompose to small molecule gases and carbon (soot) under loads of sufficient strength. We review a body of work-most of it performed at Los Alamos National Laboratory-on polymers and foams under extreme conditions. To provide some context, we begin with a brief review of basic concepts in shockwave physics, including features particular to transitions (chemical reaction or phase transition) entailing an abrupt reduction in volume. We then discuss chemical formulations and synthesis, as well as experimental platforms used to interrogate polymers under shock loading. A high-level summary of equations of state for polymers and their decomposition products is provided, and their application illustrated. We then present results including temperatures and product compositions, thresholds for reaction, wave profiles, and some peculiarities of traditional modeling approaches. We close with some thoughts regarding future work.


Novel approaches for targeting the adenosine A2A receptor.
The adenosine A2A receptor (A2AR) represents a drug target for a wide spectrum of diseases. Approaches for targeting this membrane-bound protein have been greatly advanced by new stabilization techniques. The resulting X-ray crystal structures and subsequent analyses provide deep insight to the A2AR from both static and dynamic perspectives. Application of this, along with other biophysical methods combined with fragment-based drug design (FBDD), has become a standard approach in targeting A2AR. Complementarities of in silico screening based- and biophysical screening assisted- FBDD are likely to feature in future approaches in identifying novel ligands against this key receptor. This review describes evolution of the above approaches for targeting A2AR and highlights key modulators identified. It includes a review of: adenosine receptor structures, homology modeling, X-ray structural analysis, rational drug design, biophysical methods, FBDD and in silico screening. As a drug target, the A2AR is attractive as its function plays a role in a wide spectrum of diseases including oncologic, inflammatory, Parkinson's and cardiovascular diseases. Although traditional approaches such as high-throughput screening and homology model-based virtual screening (VS) have played a role in targeting A2AR, numerous shortcomings have generally restricted their applications to specific ligand families. Using stabilization methods for crystallization, X-ray structures of A2AR have greatly accelerated drug discovery and influenced development of biophysical-in silico hybrid screening methods. Application of these new methods to other ARs and G-protein-coupled receptors is anticipated in the future.


Bleeding isolated gastric varices: a retrospective analysis.
Isolated gastric varices (IGV) are rare and are believed to be associated with left-sided portal hypertension. We studied patients presenting with bleeding from IGV and compared them with those bleeding from both esophageal and gastric varices. A retrospective analysis of 14 patients with bleeding from IGV was carried out. Portovenography findings (pattern of collateralization and natural shunts) in these patients were compared with a matched group of 69 patients with both esophageal and gastric varices. Of 14 patients with IGV, 2 had isolated splenic vein thrombosis and 12 had generalized portal hypertension. Portovenograms in 11 of the latter 12 revealed predominantly 'left-sided' collateralization in 8 patients as compared to 17 of 69 (25%) patients with esophageal and gastric varices (p = 0.004); natural shunts were seen in 6 of 11 cases and 15 of 69 (22%) patients in the two groups, respectively (p = 0.05). Abdominal devascularization operation gave good short- and long-term control of bleeding. Contrary to belief most patients with isolated gastric varices may have generalized portal hypertension rather than splenic vein obstruction as the cause and hence should be treated by a more extensive procedure than just splenectomy. The IGV could be a result of predominant collateralization to the retroperitoneal area (left-sided collateralization and natural shunts) rather than the usual pattern to the azygos system which results in esophageal varices.


[A case of urethrovaginal fistula caused by a foreign body in the vagina].
A 25-year-old woman had a urethrovaginal fistula caused by a foreign body in her vagina. The patient had sought an appointment with a gynecologist for removal of the foreign body in her vagina. However, one day before the scheduled appointment, she developed acute urinary retention and visited the emergency room of our hospital. A urinary catheter inserted into the urethra was found to come out through the vagina. The foreign body, a bottle cap made of polypropylene and measuring 48 x 28 mm in size, was removed. A cystourethroscopic examination confirmed the presence of the urethrovaginal fistula. Repeat cystourethroscopy performed after one month of conservative management showed a persistent urethrovaginal fistula, which had, in fact, increased in size as compared with that at the initial examination. Surgery was undertaken for repair of the fistula ; fistula closure was successfully accomplished by suture in three layers : urethral wall, subcutaneous tissue, and the vaginal wall. This is the second report of a urethrovaginal fistula caused by a foreign body in the vagina.


Evidence for the endogenous GTP-dependent ADP-ribosylation of the alpha-subunit of the stimulatory guanyl-nucleotide-binding protein concomitant with an increase in basal adenylyl cyclase activity in chicken spleen cell membrane.
We investigated the endogenous GTP-dependent ADP-ribosylation of the alpha-subunit of the stimulatory guanyl-nucleotide-binding protein (Gs alpha) concomitant with an increase of basal adenylyl cyclase activity in chicken spleen cell membranes. When these membranes were incubated with [adenylate-32P]NAD, there was significant incorporation of [32P]ADP-ribose into a 45-kDa acceptor protein in the membranes. This reaction was inhibited when 20 mM arginine was present during the incubation. When the membranes were incubated with unlabelled NAD, subsequent ADP ribosylation by cholera toxin was diminished significantly. Thus, chicken spleen cell membranes have the potential to endogenously ADP-ribosylate the arginine residue of Gs alpha. The endogenous ADP-ribosylation Gs alpha was enhanced by the addition of 0.1 mM GTP or 0.1 mM guanosine 5'-[gamma-thio]triphosphate (GTP[S]), but not 0.1 mM GDP, 0.1 mM ATP or 0.1 mM ADP. The endogenous GTP-dependent ADP-ribosylation of Gs alpha stimulated basal adenylyl cyclase activity. Furthermore, NAD-induced stimulation of basal adenylyl cyclase activity was suppressed, when the membranes were incubated with NAD in the presence of novobiocin, an inhibitor of arginine-specific ADP-ribosyltransferase. These data represent the first demonstration that a eukaryotic cell membrane contains an ADP-ribosyltransferase which can catalyze the endogenous GTP-dependent ADP-ribosylation of the arginine residue of Gs alpha and that this modification enhances basal adenylyl cyclase activity in the membrane. In light of this evidence, the possible control of basal adenylyl cyclase activity via endogenous GTP-dependent ADP-ribosylation in eukaryotic cells warrants further attention.


Hemodynamic force analysis is not ready for clinical trials on HFpEF.
Hemodynamic force analysis has been proposed as a novel tool for early detection of subclinical systolic dysfunction in heart failure with preserved ejection fraction (HFpEF). Here we investigated the ability of hemodynamic forces to discriminate between healthy subjects and heart failure patients with varying degrees of systolic dysfunction. We studied 34 controls, 16 HFpEF patients, and 25 heart failure patients with mid-range (HFmrEF) or reduced ejection fraction (HFrEF) using cardiac magnetic resonance with acquisition of cine images and 4D flow at 1.5 T. The Navier-Stokes equation was used to compute global left ventricular hemodynamic forces over the entire cardiac cycle. Forces were analyzed for systole, diastole, and the entire heartbeat, with and without normalization to left ventricular volume. Volume-normalized hemodynamic forces demonstrated significant positive correlation with EF (r2 = 0.47, p < 0.0001) and were found significantly lower in heart failure with reduced ejection fraction compared to controls (p < 0.0001 for systole and diastole). No difference was seen between controls and HFpEF (p > 0.34). Non-normalized forces displayed no differences between controls and HFpEF (p > 0.24 for all analyses) and did not correlate with EF (p = 0.36). Left ventricular hemodynamic force analysis, whether indexed to LV volumes or not, is not ready for clinical trials on HFpEF assessment.


[A case of metastatic Enterococcus faecalis endophthalmitis].
We reported a case of successful treatment of early-stage metastatic endophthalmitis caused by Enterococcus faecalis with vitrectomy and lensectomy. The case was a 50-year-old male with poorly controlled diabetes. Following T-tube drainage for a necrotic cholecystitis operation, he developed iridocyclitis in both eyes as well as fever. At the time of his first visit to our clinic, his right eye had already lost light perception. His left eye had visual acuity recognizing of hand movement, marked uveitis, complicated cataract, and dense vitreous opacity. As gram positive cocci were isolated from the aspirated vitreous, we conducted lensectomy and vitrectomy under irrigation of antibiotics. With systemic postoperative antibiotics and human immunoglobulin, the patient showed remarkable improvement in his ocular fundus. By 60 days after the operation, the visual acuity of his left eye recovered to 4/20. Metastatic Enterococcus faecalis endophthalmitis has almost nerve been reported in Japan. The diagnosis and treatment of this disease with a reference to the above findings were discussed.


Lack of effect of granulocyte-macrophage and granulocyte colony-stimulating factors on cultured human endothelial cells.
The hematopoietic growth factors, granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF), enhance the effector functions of mature myeloid cells, including the interaction with vascular endothelium. We examined the direct effect of recombinant human GM-CSF (rhGM-CSF) and recombinant human G-CSF (rhG-CSF) on the growth and function of cultured human umbilical vein endothelial cells (HUVEC). Endothelial cell growth supplement (ECGS) increased the proliferation of passaged and primary cells by 305% +/- 45% (mean +/- SEM, n = 5, P less than .01) over control cells at 4 days; GM-CSF and G-CSF had no effect. Endothelial cell procoagulant activity was increased after 4-hour incubation with recombinant interleukin-1 beta (IL-1 beta) 10 U/mL and recombinant tumor necrosis factor (TNF) 10 U/mL to 1,721% +/- 376% (n = 7, P less than .005) and 247% +/- 71% (n = 4) of control levels, respectively. gamma-Interferon (gamma-IFN) 50 U/mL had no direct effect of its own but was able to prime the response to IL-1 beta. There was no direct or priming effect of GM-CSF (1 ng to 1 microgram/mL) on the expression of procoagulant activity in endothelial cells. GM-CSF and G-CSF (1 ng/mL to 1 microgram/mL) had no effect on the expression of either tissue plasminogen activator (tPA) or plasminogen activator inhibitor-1 (PAI-1) by endothelial cells. The secretion of tPA by endothelial cells was increased, however, after 24-hour incubation with thrombin 4 U/mL (314% +/- 72% of control levels, n = 5, P less than .025). The production of PAI-1 was increased by TNF 200 U/mL (241% +/- 44% of control, n = 3, P less than .005), thrombin 4 U/mL (180% +/- 12% of control, n = 5, P less than .0005) and IL-1 beta 10 U/mL (275% +/- 44% of controls, n = 5, P less than .0005). In four experiments, endothelial cells showed no specific binding of 125I-GM-CSF, whereas peripheral blood (PB) neutrophils demonstrated the presence of 802 +/- 78 high-affinity receptors for GM-CSF. Thus, we found no effect of rhGM-CSF or rhG-CSF on the proliferation activities by these cells. These findings are in accordance with the lack of demonstrable receptors for GM-CSF on cultured HUVEC.


A comparison of 2,4-dichlorophenoxyacetic Acid metabolism in cultured soybean cells and in embryogenic carrot cells.
The removal or reduction in concentration of auxin is often a successful method for obtaining morphogenesis in cell cultures of higher plants, such as carrot, but not for soybean. For this reason, the metabolism of one auxin, 2,4-dichlorophenoxyacetic acid (2,4-D), was compared in both carrot and soybean cells. Whereas soybean cells conjugated a high percentage of their 2,4-D to amino acids, carrot cells contained primarily free 2,4-D. Moreover, after long-term exposure to 2,4-D, carrot cells released much more 2,4-D upon transfer to 2,4-D-free (embryogenic) medium than did soybean cells. It appears that the retention of 2,4-D by soybean cells might interfere with subsequent morphogenesis. Because no impairment of 2,4-D efflux was found with short-term exposure to radiolabeled 2,4-D, it was concluded that 2,4-D retention in soybean cells might be due to a time-dependent, metabolic process. The conjugation of 2,4-D to amino acids was shown to be one such time-dependent process. Additionally, the release of 2,4-D from the cells was shown to be due primarily to a loss of free 2,4-D and not 2,4-D-amino acid conjugates. It seems that the greater retention of 2,4-D by soybean cells upon transfer to 2,4-D-free medium is due to greater formation of 2,4-D-amino acid conjugates.


Cerebral xanthomatosis in three green water dragons (Physignathus cocincinus).
Cerebral xanthomatosis was diagnosed in three female green water dragons (Physignathus cocincinus), all of which presented with progressive neurologic signs. No antemortem evidence for xanthomatosis was identified, but on postmortem examination cholesterol granulomas, composed of cholesterol clefts surrounded by macrophages and multinucleated giant cells, were found in the forebrain of each animal and were associated with significant displacement and pressure on the adjacent brain. Although the cause of xanthomatosis in these animals is unknown, nutrition and trauma may be involved in the pathogenesis of this condition. Cerebrum, cholesterol, green water dragon, Physignathus cocincinus, xanthoma.


Glucocorticoid receptor mutants demonstrate increased motility inside the nucleus of living cells: time of fluorescence recovery after photobleaching (FRAP) is an integrated measure of receptor function.
Natural mutations of the human glucocorticoid receptor (GR) isoform alpha cause the glucocorticoid resistance syndrome. Mutant receptors may have abnormal interactions with the ligand, target DNA sequences, and/or multiple intracellular proteins, as well as aberrant nucleocytoplasmic trafficking. Using fluorescence recovery after photobleaching (FRAP) analysis, all GR pathologic mutant receptors examined, as well as 2 synthetic GR mutants lacking the activation function (AF)-1 or the ligand-binding domain (and hence the AF-2), had defective transcriptional activity and dynamic motility defects inside the nucleus of living cells. In the presence of dexamethasone, these mutants displayed a curtailed 50% recovery time (t 1/2) after photobleaching and, hence, significantly increased intranuclear motility and decreased "chromatin retention." The t 1/2 values of the mutants correlated positively with their transcriptional activities and depended on the GR domain affected. GRbeta, a natural splice variant of the GR gene, also demonstrated a shorter t 1/2 than GRalpha. The motility responsiveness of the natural and artificial mutant receptors examined, and of GRbeta, to the proteasomal inhibitor MG-132 also depended on the mutant domain. Thus, mutant glucocorticoid receptors possess dynamic motility defects in the nucleus, possibly caused by their inability to properly interact with all key partner nuclear molecules necessary for full activation of glucocorticoid-responsive genes.


Ivabradine-Flecainide as Breakthrough Drug Combination for Congenital Junctional Ectopic Tachycardia: A Case Report and Literature Review.
Congenital junctional ectopic tachycardia (CJET) is a rare tachyarrhythmia that remains difficult to manage, with suboptimal control in most cases. Here, we report literature research on the use of ivabradine in the treatment of pediatric junctional ectopic tachycardia (JET), both congenital and postoperative, and describe the successful use of ivabradine-flecainide association for CJET therapy resistant to other antiarrhythmic agents. This new drug combination was effective in completely suppressing JET. Ivabradine-flecainide combination may be considered a new therapeutic strategy of CJET with a satisfactory efficacy/tolerability ratio in patients resistant to conventional drug combinations.


A comparison of glaucoma drainage implant tube coverage.
Surgeons may use various materials, including donor sclera, dura, or pericardium grafts to cover glaucoma drainage implant tubes, prior to repositioning conjunctiva. We reviewed our experience with these materials. Sixty-four eyes with at least 24 months follow-up status post glaucoma drainage implant surgery were evaluated for signs of tube erosion, as well as patch graft thinning, after initial placement of donor sclera (23), dura (18), or pericardium (23) patch grafts. Sixty-two eyes required no intervention for conjunctival and patch graft melting with subsequent tube erosion. Three cases (two eyes) of erosion requiring reoperation (1-dura at 6 months, 1-sclera at 15 months, and in the same eye 21 months later, 1-pericardium) were noted. Significant thinning of the donor patch graft such that the tube was visible beneath intact conjunctiva ocurred in 6 out of 23 donor sclera eyes, 4 out of 18 donor dura eyes, and 6 out of 23 donor pericardium eyes. No material was more prone to melting than another. Donor sclera may be slightly more cost-efficient, but gamma-irradiated pericardium has sterility advantages.


The importance of exercise self-efficacy for clinical outcomes in pulmonary rehabilitation.
Pulmonary rehabilitation (PR) improves functional exercise capacity and health status in people with chronic obstructive pulmonary disease (COPD), although these outcomes are often not maintained following PR. Self-efficacy is a precursor to outcomes achievement, yet few studies have examined the importance of self-efficacy to outcome improvement during PR, or how it develops over time. Further, the contribution of exercise-specific self-efficacy to outcomes in PR is unknown. The aims of this study were to determine (a) whether baseline exercise self-efficacy predicts PR attendance and change in functional exercise capacity and health status over PR, and (b) if exercise self-efficacy changes with PR. Fifty-eight out of 64 patients with COPD completed PR and assessments of exercise self-efficacy (task, coping, scheduling), the 6-minute walk test (6MWT), and St. George's Respiratory Questionnaire (SGRQ) at the beginning and end of PR. Analyses were conducted to predict attendance, and change in 6MWT and SGRQ, while controlling for baseline demographic and clinical indicators. Change in 6MWT, SGRQ, and self-efficacy with PR was also examined. Clinically significant increases in the 6MWT and SGRQ were achieved with PR. Stronger task self-efficacy predicted better attendance, while stronger coping self-efficacy predicted greater 6MWT improvement. No variables predicted SGRQ change. Scheduling self-efficacy significantly improved with PR, whereas task and coping self-efficacy did not. Baseline exercise self-efficacy appears to be a determinant of rehabilitation attendance and functional exercise improvement with PR. Clinicians should evaluate and target exercise self-efficacy to maximize adherence and health outcome improvement with PR. (PsycINFO Database Record


Effect of 1-Aminocyclopropane-1-Carboxylic Acid on the Production of Ethylene in Senescing Flowers of Ipomoea tricolor Cav.
Application of 1-aminocyclopropane-1-carboxylic acid (ACC) to rib segments excised from flowers of Ipomoea tricolor Cav. resulted in the formation of C(2)H(4) in greater quantities than produced under natural conditions. The ability of ACC to enhance C(2)H(4) production was independent of the physiological age of the tissue and its capacity to synthesize C(2)H(4) without applied ACC. When ACC was fed to rib segments that had been treated with [(14)C]methionine, incorporation of radioactivity into C(2)H(4) was reduced by 80%. Aminoethoxyvinylglycine and aminooxyacetic acid inhibited C(2)H(4) production in rib segments of I. tricolor but had no effect on ACC-enhanced C(2)H(4) production. Protoplasts obtained from flower tissue of I. tricolor did not form C(2)H(4), even when incubated with methionine or selenomethionine. They produced C(2)H(4) upon incubation with ACC, however. ACC-dependent C(2)H(4) production in protoplasts was inhibited by n-propyl gallate, AgCl, CoCl(2), KCN, Na(2)S, and NaN(3). ACC-dependent C(2)H(4) synthesis in rib segments and protoplasts was dependent on O(2), the K(m) for O(2) being 1.0 to 1.4% (v/v). These results confirm the following pathway for C(2)H(4) biosynthesis in I. tricolor. methionine [selenomethionine] --> S-adenosylmethionine [selenoadenosylmethionine] --> ACC --> C(2)H(4).


Conjugated linoleic acid isomers and mammary cancer prevention.
There is increasing evidence that individual isomers of conjugated linoleic acid (CLA) may have unique biological or biochemical effects. A primary objective of this study was to determine whether there might be differences in the anticancer activity of 9,11-CLA and 10,12-CLA. This was achieved by evaluating the reduction in premalignant lesions and carcinomas in the mammary gland of rats that had been treated with a single dose of methylnitrosourea and given 0.5% of either highly purified CLA isomer in the diet. Our results showed that the anticancer efficacies of the two isomers were very similar. At 6 wk after carcinogen administration, the total number of premalignant lesions was reduced by 33-36%. At 24 wk, the total number of mammary carcinomas was reduced by 35-40%. The concentration of each CLA isomer and its respective metabolites was analyzed in the mammary fat pad. Tissue level of 10,12-CLA was much lower than that of 9,11-CLA. The pool of metabolites from each isomer was very similar between the two groups and represented only a small fraction of total conjugated diene fatty acids. Feeding of 9,11-CLA resulted in minimal changes in other unsaturated fatty acids. In contrast, feeding of 10,12-CLA produced a wider spectrum of perturbations. Small but significant increases in 16:1 and 16:2 were detected; these were accompanied by decreases in 20:2, 20:3, 20:4, 22:4, and 22:6. The above observation suggests that 10,12-CLA might be more potent than 9,11-CLA in interfering with elongation and desaturation of linoleic and linolenic acids. In summary, our study showed that, at the 0.5% dose level, the anticancer activity of 9,11-CLA and 10,12-CLA was very similar, even though accumulation of 10,12-CLA in the mammary tissue was considerably less than that of 9,11-CLA. These confounding changes of the other unsaturated fatty acids in contributing to the effect of 10,12-CLA need to be clarified.


Seasonal variations in eating disorder subtypes in Japan.
This study estimated the degree of seasonal changes in eating disorder subtypes in Japan. A Japanese version of the Seasonal Pattern Assessment Questionnaire (SPAQ) was mailed to 228 outpatients with eating disorders in Kyoto (latitude 35 degrees 01' N). Ninety responses were analyzed. The mean of the global seasonality score (GSS) in the SPAQ and the comorbidity rate of seasonal affective disorder (SAD) and subsyndromal-SAD in each subtype were used as indices of seasonality. The mean (8.2) of the GSS of the bulimia nervosa (BN) group and the sum (43%) of the comorbidity rates of SAD and subsyndromal-SAD in the BN group were significantly higher than those in the nonclinical comparison group. The seasonality of BN in Kyoto is the highest among eating disorder subtypes, although lower than that in North America. This finding suggests that light therapy could be useful for Japanese BN patients.


Purification and characterization of glutamine synthetase from Clostridium pasteurianum.
Glutamine synthetase from Clostridium pasteurianum grown on molasses as the sole carbon source and ammonium chloride as the nitrogen source has been purified to homogeneity (45-fold) with 32% recovery. The procedure involves ammonium sulfate precipitation and chromatography on a combined Sepharose 4B/DEAE-Sephadex A-50 column. The purified enzyme being very unstable was stabilized by the addition of 25% (v/v) glycerol. The enzyme has an unusually high molecular weight of 1 X 10(6) and 20 subunits of Mr 50 000 each, as determined by gel filtration and sodium dodecyl sulfate gel electrophoresis, respectively. It has an absorption maximum at 280 nm and a fluorescence emission maximum at 380 nm when excited at 280 nm. Its substrate binding pattern as studied by fluorescence quenching studies is different from that of the Escherichia coli enzyme. Both the gamma-glutamyltransferase and synthetase activities reside in the same protein as the ratio of the two activities at each step of purification remains constant and the enzyme exhibits optimal transferase and synthetase activities at the same pH (7.2) and temperature (50 degrees C). The thermal stabilities of both activities were also similar, and decay of both the activities at 50 degrees C ran parallel. The enzyme shows stabilization by substrates, as L-glutamate, Mg2+, and ATP + Mg2+ protected both the synthetase and gamma-glutamyltransferase activities against thermal inactivation. Storage in 25% (v/v) glycerol enhanced the thermal stability of glutamine synthetase. Metal ion requirement and substrate specificity of the enzyme have been examined. Maximum synthetase activity occurs when [Mg2+]: [ATP] = 2. The Km app values are as follows (in parentheses): ATP (0.34 mM), NH2OH (0.4 mM in the synthetase reaction and 4.1 mM in the transferase reaction), glutamine (14.7 mM), ADP (3.8 X 10(-4) mM), arsenate (2.5 mM), and L-glutamate (3.4 mM, 22.2 mM). The enzyme exhibits negative cooperativity in the binding of glutamate. Amino acids such as L-serine, glycine, L-alanine, and L-aspartic acid inhibit the enzyme.


Effects of femoral arteriography and low osmolar contrast agents on renal function.
The contrast media-induced renal effects of 3 different low osmolar contrast agents were prospectively evaluated with creatinine, beta-2-microglobulin, and urea in serum before and after femoral arteriography in 110 consecutive patients. Forty-two patients (38%) had at least one of the 2 major risk factors for contrast media-induced nephropathy; diabetes mellitus or renal dysfunction. Six patients (5%) had both risk factors. There were statistically significant increases of S-creatinine (6.5%, p < 0.001) and S-beta-2-microglobulin (7.4%, p < 0.001) but no increase of S-urea, in the total patient material. This effect was independent of preexaminatory renal function, the presence of diabetes mellitus and type of low osmolar contrast agent used in these patients who were properly hydrated before the examination. The amount of administered contrast medium did not correlate with the degree of contrast agent-induced renal function impairment.


[Construction of nsdAmgh gene disruption mutant in Strempomyces roseoflavus Men-myco-93-63].
Insertional mutagenesis is a widely used method to determine the function(s) of a gene. To study the function(s) of the gene nsdAmgh in Streptomyces roseoflavus, a homologous recombination vector pSRNA2500 was structured in this paper. The recombination donor vector was then transformed into Strempomyces roseoflavus strain Men-myco-93-63 by conjugative transfer. The transformants were subjected to selection under the pressure of high temperature and appropriate antibiotics. As a result, several disrupted mutants of nsdAmgh gene, with a phenotype of Am(s)Km(r), were isolated and verified using PCR and Dot-blotting and Southern blotting hybridization methods. Functional analysis showed that the disrupted mutants of nsdAmgh had a two-fold higher inhibition against Verticillium dahlia Kleb than that of the wild strain Men-myco-93-63, which all will provide a new study route for future research about positive and negative regulator in Men-myco-93-63.


Galanin: a hypothalamic-hypophysiotropic hormone modulating reproductive functions.
Galanin (GAL) is widely distributed in the peripheral and the central nervous systems. In the brain, the highest GAL concentrations are observed within the hypothalamus and, particularly, in nerve terminals of the median eminence. This location, as well as GAL actions on prolactin, growth hormone, luteinizing hormone (LH), and LH-releasing hormone (LHRH) secretion, suggest the possibility that GAL may act as a putative hypothalamic-hypophysiotropic hormone. To establish this, GAL and LHRH levels were measured in hypophyseal portal plasma samples using specific radioimmunoassays. Rat galanin (rGAL) concentrations in portal blood were approximately 7-fold higher than those observed in peripheral plasma in male and female (estrus, diestrus) rats, indicating an active secretory process of rGAL into the portal vasculature. Frequent (10 min) sampling revealed that rGAL and LHRH were secreted into the portal circulation in a pulsatile manner with a pulse frequency of one pulse per hour. Interestingly, both hormone series depicted a high degree of coincident episodes. In fact, the probability of random coincidence, calculated by the algorithm HYPERGEO, was less than 0.01. Moreover, the retrograde tracer Fluoro-Gold, when given systemically, was taken up by GAL neurons in the hypothalamus, including a subset of neurons expressing rGAL and LHRH, strengthening the notion of the existence of a GAL neuronal system connected to the hypophyseal portal circulation. These observations reinforce the concept that GAL regulates pituitary hormone secretion. To analyze this in further detail, the effects of rGAL on LH secretion were evaluated under basal and stimulated conditions. rGAL induced a small but dose-dependent increase in LH secretion from cultured, dispersed pituitary cells. Interestingly, rGAL enhanced the ability of LHRH to stimulate LH release. The tight link between GAL and LHRH neuronal systems is strengthened by the observation that during the estrous cycle of the rat, rGAL and LHRH contents in the median eminence show an identical profile (r = 1.00). These data indicate that GAL should be considered as a hypothalamic-hypophysiotropic hormone and as an important neuromodulator of LHRH secretion and action. The colocalization and cosecretion of GAL and LHRH and the cooperative action at the level of the anterior pituitary afford important evidence for the functional significance of coexistence of neurotransmitters in neurons of the central nervous system.