CB2 Agonist (AM1241) Improving Effect on Ovalbumin-Induced Asthma in Rats.
Asthma is a disease characterized by spontaneous contraction of the airways in response to a wide variety of endogenous and exogenous stimuli. Many asthma models are used to mimic the human asthma model in the literature. In order to better understand the role of the cannabinoid (CB) 2 receptor in the ovalbumin (OVA)-induced asthma model, a combination of both selective CB2 agonist (AM1241) and antagonist (AM630) was used to improve inflammatory hypersensitivity and edema in rats. In the present study, it was found that OVA decreased body weight (p < 0.05), increased lung weights (p < 0.05), increased diastolic and systolic blood pressure (p < 0.001), and caused irregularity in pulmonary functions (p < 0.001). Moreover, CB2 agonist was found not to reduce body weight, cause blood pressure and respiratory irregularities (p < 0.05). OVA led to increase in IgE, TNF-α, IL-4, MDA level (p < 0.001), and total WBC count (p < .05). CB2 treatment caused to reduce the number of total WBC and the level of total protein in BALF, to hinder to increase level of MDA, IgE, TNF-α, and IL-4 (p < 0.05) in BALF or serum or lung tissue. But CB2-antagonist treatment prevented the protective effect of CB2 agonist. The aim of this study was to study the role of the CB2 receptor in the OVA induced asthma model, to improve inflammatory hypersensitivity, and edema in the rats. The results suggested that CB2 agonist administration to OVA induced asthmatic rats via anti-asthmatic potential through inhibition of parameters such as IgE, IL-4, TNF-α, microvascular escape, and oxidative stress.


The association between spouses' earnings and trends in income inequality in Brazil (1993-2015).
In this paper, we investigate how the diffusion of dual-earner couples has affected aggregate levels of inequality in Brazil. More specifically, we analyze trends in the association between spouses' earnings and assess their implications for earnings inequality among couples from 1993 to 2015. For this purpose, we use log-linear models to distinguish three components of the association between spouses' earnings: a) the correlation between spouses' earnings among dual-earner couples; b) the relationship between husbands' earnings and wives' labor force participation; and c) the proportion of dual-earner couples. Counterfactual simulations allow us to estimate how inequality would change if the trends in the association between spouses' earnings and each of its components had been different. We show that changes in the gradient of wives' employment and compositional effects related to the increasing prevalence of dual-earner couples contributed to limit the decline in inequality over the studied period.


Differentiation between intra-axial metastatic tumor progression and radiation injury following fractionated radiation therapy or stereotactic radiosurgery using MR spectroscopy, perfusion MR imaging or volume progression modeling.
To determine the accuracy of magnetic resonance spectroscopy (MRS), perfusion MR imaging (MRP), or volume modeling in distinguishing tumor progression from radiation injury following radiotherapy for brain metastasis. Twenty-six patients with 33 intra-axial metastatic lesions who underwent MRS (n=41) with or without MRP (n=32) after cranial irradiation were retrospectively studied. The final diagnosis was based on histopathology (n=4) or magnetic resonance imaging (MRI) follow-up with clinical correlation (n=29). Cho/Cr (choline/creatinine), Cho/NAA (choline/N-acetylaspartate), Cho/nCho (choline/contralateral normal brain choline) ratios were retrospectively calculated for the multi-voxel MRS. Relative cerebral blood volume (rCBV), relative peak height (rPH) and percentage of signal-intensity recovery (PSR) were also retrospectively derived for the MRPs. Tumor volumes were determined using manual segmentation method and analyzed using different volume progression modeling. Different ratios or models were tested and plotted on the receiver operating characteristic curve (ROC), with their performances quantified as area under the ROC curve (AUC). MRI follow-up time was calculated from the date of initial radiotherapy until the last MRI or the last MRI before surgical diagnosis. Median MRI follow-up was 16 months (range: 2-33). Thirty percent of lesions (n=10) were determined to be radiation injury; 70% (n=23) were determined to be tumor progression. For the MRS, Cho/nCho had the best performance (AUC of 0.612), and Cho/nCho >1.2 had 33% sensitivity and 100% specificity in predicting tumor progression. For the MRP, rCBV had the best performance (AUC of 0.802), and rCBV >2 had 56% sensitivity and 100% specificity. The best volume model was percent increase (AUC of 0.891); 65% tumor volume increase had 100% sensitivity and 80% specificity. Cho/nCho of MRS, rCBV of MRP, and percent increase of MRI volume modeling provide the best discrimination of intra-axial metastatic tumor progression from radiation injury for their respective modalities. Cho/nCho and rCBV appear to have high specificities but low sensitivities. In contrast, percent volume increase of 65% can be a highly sensitive and moderately specific predictor for tumor progression after radiotherapy. Future incorporation of 65% volume increase as a pretest selection criterion may compensate for the low sensitivities of MRS and MRP.


A clinical practice review of crying research.
The goal of this clinical practice review is to assess the current state of the theoretical and empirical literature on emotional crying (i.e., crying in response to an emotional stimulus), a topic that has received surprisingly limited attention of behavioral scientists and clinicians. Although the empirical research on emotional crying remains in a nascent state, we draw upon the existing scientific knowledge to provide preliminary suggestions for clinicians on how to interpret and respond to crying in clinical contexts. We also identify research gaps and provide recommendations for further research to improve our understanding of this intriguing and still poorly understood human behavior. We suggest that a better understanding of individual differences in crying behavior and the postulated intraindividual and interindividual functions of crying is of critical importance for clinicians, given its frequent occurrence and notable associations with emotional and social functioning. An improved characterization of this important phenomenon will lead to improvements in clinical assessment, treatment planning, and psychotherapy interventions. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Sensitivity and selectivity of neurons in auditory cortex to the pitch, timbre, and location of sounds.
We are able to rapidly recognize and localize the many sounds in our environment. We can describe any of these sounds in terms of various independent "features" such as their loudness, pitch, or position in space. However, we still know surprisingly little about how neurons in the auditory brain, specifically the auditory cortex, might form representations of these perceptual characteristics from the information that the ear provides about sound acoustics. In this article, the authors examine evidence that the auditory cortex is necessary for processing the pitch, timbre, and location of sounds, and document how neurons across multiple auditory cortical fields might represent these as trains of action potentials. They conclude by asking whether neurons in different regions of the auditory cortex might not be simply sensitive to each of these three sound features but whether they might be selective for one of them. The few studies that have examined neural sensitivity to multiple sound attributes provide only limited support for neural selectivity within auditory cortex. Providing an explanation of the neural basis of feature invariance is thus one of the major challenges to sensory neuroscience obtaining the ultimate goal of understanding how neural firing patterns in the brain give rise to perception.


GADD45 proteins: central players in tumorigenesis.
The Growth Arrest and DNA Damage-inducible 45 (GADD45) proteins have been implicated in regulation of many cellular functions including DNA repair, cell cycle control, senescence and genotoxic stress. However, the pro-apoptotic activities have also positioned GADD45 as an essential player in oncogenesis. Emerging functional evidence implies that GADD45 proteins serve as tumor suppressors in response to diverse stimuli, connecting multiple cell signaling modules. Defects in the GADD45 pathway can be related to the initiation and progression of malignancies. Moreover, induction of GADD45 expression is an essential step for mediating anti-cancer activity of multiple chemotherapeutic drugs and the absence of GADD45 might abrogate their effects in cancer cells. In this review, we present a comprehensive discussion of the functions of GADD45 proteins, linking their regulation to effectors of cell cycle arrest, DNA repair and apoptosis. The ramifications regarding their roles as essential and central players in tumor growth suppression are also examined. We also extensively review recent literature to clarify how different chemotherapeutic drugs induce GADD45 gene expression and how its up-regulation and interaction with different molecular partners may benefit cancer chemotherapy and facilitate novel drug discovery.


Alpha-smooth-muscle actin in and contraction of porcine dental pulp cells.
The finding of expression of a muscle actin isoform, alpha-smooth-muscle actin (SMA), by the stromal cells of several tissues prompted this study of SMA expression by cells derived from the porcine dental pulp. The SMA content of the cells increased with time in culture. These SMA-containing cells were found to have the capability to contract a collagen-glycosaminoglycan analog of extracellular matrix in vitro.


Phase II trial of imatinib mesylate in patients with recurrent platinum- and taxane-resistant low-grade serous carcinoma of the ovary, peritoneum, or fallopian tube.
To evaluate the efficacy and tolerability of imatinib mesylate in patients with recurrent low-grade serous carcinoma (LGSC) of the ovary, peritoneum, or fallopian tube. This open-label, single-institution phase II trial enrolled patients with platinum-resistant LGSC who had measurable disease, had received up to 4 platinum- and/or taxane-containing chemotherapy regimens, and had been previously screened for at least one imatinib targeted biomarker (c-kit, platelet-derived growth factor receptor [PDGFR]-β, or bcr-abl). Imatinib (600 mg) was administered daily for one 6-week course and continued in the absence of toxicity and disease progression. Thirteen patients were enrolled; 12 were evaluable for toxicity, and 11 were evaluable for response. A total of 17 courses were administered (median, 1 course; range, 1-5 courses). Complete or partial responses were not observed. One patient had stable disease for 7.3 months. c-Kit, bcr-abl, or PDGFR-β were present in 48%, 77%, and 100% of patients, respectively. No correlation between best response (stable disease) and the presence of imatinib-targeted biomarkers was observed. Adverse events included grade 3 skin rash in one patient leading to discontinuation of the drug, and grade 3 febrile neutropenia and grade 2 weight gain in two patients leading to dose reductions. The most common grade 1 or 2 toxicities were fatigue (66%), nausea/vomiting (66%), and diarrhea (41%); grade 3 toxicities included skin rash and granulocytopenia events. No grade 4 or 5 toxicities were observed. The median progression-free survival time was 1.3 months (95% CI, 1.27, 1.40 months), and the median overall survival time was 14.9 months (95% CI, 11.0, 18.9 months). Imatinib is well-tolerated but has no activity in patients with platinum- and taxane-resistant LGSC or the ovary, peritoneum, or fallopian tube.


Unsaturation of the membrane lipids of chloroplasts stabilizes the photosynthetic machinery against low-temperature photoinhibition in transgenic tobacco plants.
Using tobacco plants that had been transformed with the cDNA for glycerol-3-phosphate acyltransferase, we have demonstrated that chilling tolerance is affected by the levels of unsaturated membrane lipids. In the present study, we examined the effects of the transformation of tobacco plants with cDNA for glycerol-3-phosphate acyltransferase from squash on the unsaturation of fatty acids in thylakoid membrane lipids and the response of photosynthesis to various temperatures. Of the four major lipid classes isolated from the thylakoid membranes, phosphatidylglycerol showed the most conspicuous decrease in the level of unsaturation in the transformed plants. The isolated thylakoid membranes from wild-type and transgenic plants did not significantly differ from each other in terms of the sensitivity of photosystem II to high and low temperatures and also to photoinhibition. However, leaves of the transformed plants were more sensitive to photoinhibition than those of wild-type plants. Moreover, the recovery of photosynthesis from photoinhibition in leaves of wild-type plants was faster than that in leaves of the transgenic tobacco plants. These results suggest that unsaturation of fatty acids of phosphatidylglycerol in thylakoid membranes stabilizes the photosynthetic machinery against low-temperature photoinhibition by accelerating the recovery of the photosystem II protein complex.


Influences of Different Operative Methods on the Recurrence Rate of Non-Muscle-Invasive Bladder Cancer.
To compare the influence of three operative approaches [transurethral en bloc resection of bladder tumor by pin-shaped electrode (pin-ERBT), transurethral resection of bladder tumor (TURBT) and transurethral holmium laser resection of bladder tumor (HoLRBT)] on the recurrence rate of non-muscle-invasive bladder cancer (NMIBC) at low dimension (i.e. diameter below 3 cm). A retrospective analysis was conducted for a total of 115 patients affected by solitary NMIBC, with a diameter <3 cm, who were submitted to operation between March 2013 to May 2017. The patients were divided according to the operative method applied (pin-ERBT, TURBT and HoLRBT groups, respectively). The 2-year recurrence rate was compared among the three groups, and multivariat Cox hazard model analysis was applied to analyze the influencing factor(s) for postoperative recurrence. The 2-year recurrence rate was 10.0% in ERBT, 38.5% in TURBT and 40.0% in HoLRBT group, with a significant difference (P =0.014). According to the Cox hazard model analysis, age(HR=1.058, 95% CI: 1.019~1.098,P=0.003), operative method(HR=2.974,6.508, 95% CI: 0.862~10.255,1.657~25.566, P=0.023), smoking(HR=2.399, 95% CI: 1.147~5.017, P=0.020) and pathological grade(HR=2.012,95% CI: 1.279~3.165, P=0.002) were risk factors for postoperative recurrence of bladder cancer. Pin-ERBT can prominently decrease the postoperative recurrence rate of solitary NMIBC with a diameter <3 cm.


Early Childhood Caries Intervention in Aboriginal Australian Children: A Cross-in Randomized Trial.
Early childhood caries (ECC) is the strongest predictor of dental caries in adulthood. Indigenous children have higher levels of ECC compared with non-Indigenous children. The study aimed to estimate the efficacy of an ECC intervention among Aboriginal Australian children. Baby Teeth Talk was an outcome assessor-blinded, closed-cohort cross-in randomized controlled trial conducted in South Australia, Australia. We randomly allocated 448 women pregnant with an Aboriginal child to either an immediate (II) or delayed (DI) intervention group between January 2011 and May 2012. The immediate intervention comprised (1) provision of dental care to mothers during pregnancy; (2) application of fluoride varnish to teeth of children at ages 6, 12; and 18 mo; (3) motivational interviewing delivered in conjunction; and (4) anticipatory guidance. Mothers/children in the DI group received the same intervention commencing when the child was 24 mo of age. The outcomes were assessed by the number of decayed, missing, and filled teeth (dmft) in children aged 24, 36, and 60 mo. Regression-based approaches with generalized linear mixed effect (log-Poisson) model characterized the mean dmft to estimate risk ratios (RRs) and 95% confidence intervals (95% CIs). A total of 223 participants were randomly allocated to the II group and 225 to the DI group. Three hundred sixty-five children (178 II, 187 DI) received at least 1 dental clinical examination at 24, 36, and 60 mo of follow-up. The mean dmft was lower in the II group (0.48, 1.32, and 2.06) than in the DI group (0.82, 1.90, and 3.29) at each time point, respectively. The direct ECC intervention effect was to prevent/decrease dental decay experience (dmft) occurrence by 84% (RR = 1.84, 95% CI: 1.20-2.48) after adjusting for all covariates. Our analysis indicated that the time-varied ECC intervention had immediate and longer-term effects on the prevention of ECC among Indigenous Australian children. The study aimed to estimate the efficacy of an early childhood caries (ECC) intervention among Aboriginal Australian children. The findings indicated that the culturally appropriate ECC intervention is effective for the preventive of ECC and can be used by clinicians, educators, and policy makers when planning an ECC intervention, so as to prevent and reduce ECC and meet identified oral health needs across the Australian population, which is important for preventive dental care.


Clinical outcomes of hepatitis C treatment before and after kidney transplantation and its impact on time to transplant: A multicenter study.
Waitlist time for kidney transplantation is long but may be shortened with the utilization of hepatitis C positive allografts. We retrospectively reviewed the course of 36 hepatitis C positive patients awaiting kidney transplantation at 2 large centers within the same health system, with near-identical care delivery models with the exception of timing of hepatitis C treatment, to determine the impact of timing of hepatitis C treatment on access to transplant, waitlist time, and treatment efficacy and tolerability. The majority of patients had hepatitis C genotype 1a or 1b, and all received direct acting antiviral therapy with 100% treatment response. One patient underwent transplantation in the pretransplant treatment group. The 1-year transplantation rate was 12.5% vs 67.9% (P = .0013) in those treated posttransplantation. The median waitlist time in the posttransplant group was 122 (interquartile range [IQR] 21.5, 531.0) days, which was significantly shorter than the center's regional and national wait time. Pathologic review revealed no difference in allograft quality. Overall treatment related adverse events were not different between the 2 groups. A strategy of posttransplant hepatitis C treatment increased access to transplant and reduced waitlist time. Delaying treatment until after transplant did not appear to adversely affect recipients' kidney allograft or overall survival.


Formation of silica aggregates in sorghum root endodermis is predetermined by cell wall architecture and development.
Deposition of silica in plant cell walls improves their mechanical properties and helps plants to withstand various stress conditions. Its mechanism is still not understood and silica-cell wall interactions are elusive. The objective of this study was to investigate the effect of silica deposition on the development and structure of sorghum root endodermis and to identify the cell wall components involved in silicification. Sorghum bicolor seedlings were grown hydroponically with (Si+) or without (Si-) silicon supplementation. Primary roots were used to investigate the transcription of silicon transporters by quantitative RT-PCR. Silica aggregation was induced also under in vitro conditions in detached root segments. The development and architecture of endodermal cell walls were analysed by histochemistry, microscopy and Raman spectroscopy. Water retention capability was compared between silicified and non-silicified roots. Raman spectroscopy analyses of isolated silica aggregates were also carried out. Active uptake of silicic acid is provided at the root apex, where silicon transporters Lsi1 and Lsi2 are expressed. The locations of silica aggregation are established during the development of tertiary endodermal cell walls, even in the absence of silicon. Silica aggregation takes place in non-lignified spots in the endodermal cell walls, which progressively accumulate silicic acid, and its condensation initiates at arabinoxylan-ferulic acid complexes. Silicification does not support root water retention capability; however, it decreases root growth inhibition imposed by desiccation. A model is proposed in which the formation of silica aggregates in sorghum roots is predetermined by a modified cell wall architecture and takes place as governed by endodermal development. The interaction with silica is provided by arabinoxylan-ferulic acid complexes and interferes with further deposition of lignin. Due to contrasting hydrophobicity, silicification and lignification do not represent functionally equivalent modifications of plant cell walls.


Identification of the Function of the Pathogenesis-Related Protein GmPR1L in the Resistance of Soybean to Cercospora sojina Hara.
Pathogenesis-related proteins, often used as molecular markers of disease resistance in plants, can enable plants to obtain systemic resistance. In this study, a gene encoding a pathogenesis-related protein was identified via RNA-seq sequencing analysis performed at different stages of soybean seedling development. Because the gene sequence showed the highest similarity with PR1L sequence in soybean, the gene was named GmPR1-9-like (GmPR1L). GmPR1L was either overexpressed or silenced in soybean seedlings through Agrobacterium-mediated transformation to examine the resistance of soybean to infection caused by Cercospora sojina Hara. The results revealed that GmPR1L-overexpressing soybean plants had a smaller lesion area and improved resistance to C. sojina infection, whereas GmPR1L-silenced plants had low resistance to C. sojina infection. Fluorescent real-time PCR indicated that overexpression of GmPR1L induced the expression of genes such as WRKY, PR9, and PR14, which are more likely to be co-expressed during C. sojina infection. Furthermore, the activities of SOD, POD, CAT, and PAL were significantly increased in GmPR1L-overexpressing soybean plants after seven days of infection. The resistance of the GmPR1L-overexpressing lines OEA1 and OEA2 to C. sojina infection was significantly increased from a neutral level in wild-type plants to a moderate level. These findings predominantly reveal the positive role of GmPR1L in inducing resistance to C. sojina infection in soybean, which may facilitate the production of improved disease-resistant soybean cultivars in the future.


Induction of Pro-Apoptotic Endoplasmic Reticulum Stress in Multiple Myeloma Cells by NEO214, Perillyl Alcohol Conjugated to Rolipram.
Despite the introduction of new therapies for multiple myeloma (MM), many patients are still dying from this disease and novel treatments are urgently needed. We have designed a novel hybrid molecule, called NEO214, that was generated by covalent conjugation of the natural monoterpene perillyl alcohol (POH), an inducer of endoplasmic reticulum (ER) stress, to rolipram (Rp), an inhibitor of phosphodiesterase-4 (PDE4). Its potential anticancer effects were investigated in a panel of MM cell lines. We found that NEO214 effectively killed MM cells in vitro with a potency that was over an order of magnitude stronger than that of its individual components, either alone or in combination. The cytotoxic mechanism of NEO214 involved severe ER stress and prolonged induction of CCAAT/enhancer-binding protein homologous protein (CHOP), a key pro-apoptotic component of the ER stress response. These effects were prevented by salubrinal, a pharmacologic inhibitor of ER stress, and by CHOP gene knockout. Conversely, combination of NEO214 with bortezomib, a drug in clinical use for patients with MM, resulted in synergistic enhancement of MM cell death. Combination with the adenylate cyclase stimulant forskolin did not enhance NEO214 impact, indicating that cyclic adenosine 3',5'-monophosphate (AMP) pathways might play a lesser role. Our study introduces the novel agent NEO214 as a potent inducer of ER stress with significant anti-MM activity in vitro. It should be further investigated as a potential MM therapy aimed at exploiting this tumor's distinct sensitivity to ER stress.


The role of octopamine and tyramine in Drosophila larval locomotion.
The characteristic crawling behavior of Drosophila larvae consists of a series of rhythmic waves of peristalsis and episodes of head swinging and turning. The two biogenic amines octopamine and tyramine have recently been shown to modulate various parameters of locomotion, such as muscle contraction, the time spent in pausing or forward locomotion, and the initiation and maintenance of rhythmic motor patterns. By using mutants having altered octopamine and tyramine levels and by genetic interference with both systems we confirm that signaling of these two amines is necessary for larval locomotion. We show that a small set of about 40 octopaminergic/tyraminergic neurons within the ventral nerve cord is sufficient to trigger proper larval locomotion. Using single-cell clones, we describe the morphology of these neurons individually. Given various potential roles of octopamine and tyramine in the larval brain, such as locomotion, learning and memory, stress-induced behaviors or the regulation of the energy state, functions that are often not easy to discriminate, we dissect here for the first time a subset of this complex circuit that modulates specifically larval locomotion. Thus, these data will help to understand-for a given neuronal modulator-how specific behavioral functions are executed within distinct subcircuits of a complex neuronal network.


Aspects on the Ecology of Phlebotomine Sand Flies (Diptera: Psychodidae) From Guaraí, State of Tocantins, Brazil, Endemic Area for American Cutaneous Leishmaniasis.
In Brazil, American cutaneous leishmaniasis (ACL) ecology involves a diversity of Leishmania species transmitted by different sand fly species. Workers involved in agricultural activities are those mainly affected by ACL in some regions from Tocantins State (TO), Brazil, where the disease can be established in new settlements. The objective of this study was to examine the seasonal and hourly frequency of sand fly species, focusing on the potential vectors of ACL, in a settlement in Guaraí (TO), an ACL transmission area. Sand flies were captured in forested area close to Pedra Branca Agricultural Project settlement, from March 2006 until December 2007, using Shannon trap. Monthly captures were made from 06:00 p.m. to 10:00 p.m., and 24-h captures were done twice per semester, from 06:00 a.m. to 06:00 a.m. A total of 10,089 specimens from 30 species were identified. Psychodopygus complexus Mangabeira, Psychodopygus llanosmartinsi Fraiha & Ward, and Nyssomyia antunesi Coutinho were the most abundant species. Nyssomyia antunesi was more frequent during the dry period, whereas Ps. complexus and Ps. llanosmartinsi had high frequencies during the rainy season. Precipitation was positively correlated with Ps. complexus and Ps. llanosmartinsi abundance, and negatively correlated with Ny. antunesi During 24-h captures, the majority of specimens were captured during the night followed by a decrease at dawn. The behavior and previous finding of natural infection by Leishmania (Viannia) braziliensis of Ps. complexus led us to the conclusion that this species can be a potential vector of L. (V.) braziliensis during the rainy season in Guaraí.


Apical extrusion of sodium hypochlorite using different root canal irrigation systems.
Root canal irrigation carries a risk of extrusion of irrigant into the periapical tissues. The objective of this study was to compare different irrigation systems in matched pairs of teeth prepared to an apical size of 35.06 and 50.06 by measuring the frequency and extent of apical extrusion of sodium hypochlorite (NaOCl) into a simulated periapical environment. The null hypothesis was tested that there is no difference between systems. Bilaterally matched pairs (n = 10) of single-canal extracted human anterior teeth were instrumented to an apical size of either 35.06 or 50.06. Teeth were embedded in a gel containing the pH-sensitive dye M-cresol purple that changes from yellow at pH 7.4 to purple at pH 9. Root canals were irrigated with 6% NaOCl (pH 11) by using EndoActivator (EA), EndoVac (EV), Rispi-Sonic/MicroMega 1500 (MM), passive ultrasonic irrigation (PUI), and syringe irrigation with a slot-tipped needle (SN), so that each tooth underwent all irrigation procedures in a randomized crossover design. Apical extrusion was evaluated by image analyses. The frequency of extrusion was less in teeth with apical preparation size 35.06 (36%) compared with 50.06 (60%) (P = .014) and was dependent on the irrigation system in 35.06 (P = .039) but not 50.06 groups. In the 35.06 group the frequency of extrusion was less for EV than for MM and SN (both P = .029). The extent of extrusion was less for MM compared with PUI (P = .024) and SN (P = .046) in the 35.06 group and greater for SN compared with all other systems in the 50.06 group (P < .05). The null hypothesis was rejected. The frequency of apical extrusion of NaOCl was dependent on the type of root canal irrigation system and apical preparation size. The extent of extrusion depended on the irrigation system, with syringe and slotted-needle irrigation resulting in the greatest extent of extrusion.


[Oxygen-lactated Ringer's solution for surgical patients].
The effects of intravenous infusion of the lactated Ringer's solution inflated with oxygen (Oxygen-LR) and lactated Ringer's solution (LR) on oxygenation of blood were studied in 23 patients undergoing abdominal surgery under epidural anesthesia. LR 1000 ml was infused with inflated oxygen at 1L/min in 30 minutes in the oxygen-LR group (n = 12). The partial pressure of oxygen-LR was increased to 600.45 +/- 78.78 from 129.4 +/- 9.69 mmHg. LR 1000 ml was infused in 30 minutes in the control group (n = 11). There was no significant difference in PaO2, PaCO2, SatO2, A-aDO2 during and 30 minutes after infusion between the oxygen-LR and control groups. We believe that oxygen-LR is of no clinical value because of its low ability is of no carrying oxygen.


Prevalence, Virulence Genes, Antimicrobial Susceptibility, and Genetic Diversity of Bacillus cereus Isolated From Pasteurized Milk in China.
Bacillus cereus is a common and important food-borne pathogen that can be found in various food products. Due to low-temperature sterilization for a short period of time, pasteurization is not sufficient for complete elimination of B. cereus in milk, thereby cause severe economic loss and food safety problems. It is therefore of paramount importance to perform risk assessment of B. cereus in pasteurized milk. In this study, we isolated B. cereus from pasteurized milk samples in different regions of China, and evaluated the contamination situation, existence of virulence genes, antibiotic resistance profile and genetic polymorphism of B. cereus isolates. Intriguingly, 70 samples (27%) were found to be contaminated by B. cereus and the average contamination level was 111 MPN/g. The distribution of virulence genes was assessed toward 10 enterotoxigenic genes (hblA, hblC, hblD, nheA, nheB, nheC, cytK, entFM, bceT, and hlyII) and one emetic gene (cesB). Forty five percent strains harbored enterotoxigenic genes hblACD and 93% isolates contained nheABC gene cluster. The positive rate of cytK, entFM, bceT, hlyII, and cesB genes were 73, 96, 75, 54, and 5%, respectively. Antibiotic susceptibility assessment showed that most of the isolates were resistant to β-lactam antibiotics and rifampicin, but susceptible to other antibiotics such as ciprofloxacin, gentamicin and chloramphenicol. Total multidrug-resistant population was about 34%. In addition, B. cereus isolates in pasteurized milk showed a high genetic diversity. In conclusion, our findings provide the first reference on the prevalence, contamination level and characteristics of B. cereus isolated from pasteurized milk in China, suggesting a potential high risk of B. cereus to public health and dairy industry.


[Effect of diadenosine oligophosphates (Ap4A and Ap3A) and their phosphonate analogs on catalytic properties of phenylalanyl-tRNA synthetase from E. coli].
The influence of P1,P3-bis(5'-adenosyl)triphosphate (Ap3A), P1,P4-bis(5'-adenosyl)tetraphosphate (Ap4A) and its analogues, containing a residue of methylenediphosphonic acid in various positions of the oligophosphate chain, on the reactions catalysed by phenylalanyl-tRNA synthetase from E. coli MRE-600 has been studied. The compounds do not affect significantly the rate of ATP-[32P]PPi-exchange nor maintain this reaction in the absence of ATP. The diadenosineoligophosphates are shown to be noncompetitive inhibitors of ATP in the tRNA aminoacylation by phenylalanine (for Ap4A Ki = 1,45.10(-3) M). The phosphonate analogues of Ap4A inhibit the synthesis of Ap3A depending on their structure. The conclusion is thus drawn that the E. coli MRE-600 phenylalanyl-tRNA synthetase does not interact property with Ap4A and its phosphonate analogues.


Microencapsulation of beta-galactosidase with fatty acid esters.
The present study was carried out to find an appropriate coating material to microencapsulate lactase and the conditions under which it could be used. As a coating material, medium-chain triacylglycerol (MCT) and polyglycerol monostearate (PGMS) were superior. Three different percentages of microcapsules (2, 4, and 6%) were added and subdivided into three groups by washing time (0, 1, and 2). The highest efficiency of microencapsulation was found in the ratio of 15:1 as coating to core material with both MCT (94.9%) and PGMS (72.8%). Lactose content was not significantly changed up to 12 d with MCT and up to 8 d with PGMS with 2% of twice washed microcapsule addition. Also, sweetness was not significantly increased in storage compared with that of market milk. The present study provides evidence that emulsifiers can be used as an effective coating material to microencapsulate lactase. In addition, these results suggest that acceptable milk products can be prepared with microencapsulated lactase.


Gene expression profiling of flaxseed in mouse lung tissues-modulation of toxicologically relevant genes.
Flaxseed (FS), a nutritional supplement consisting mainly of omega-3 fatty acids and lignan phenolics has potent anti-inflammatory, anti-fibrotic and antioxidant properties. The usefulness of flaxseed as an alternative and complimentary treatment option has been known since ancient times. We have shown that dietary FS supplementation ameliorates oxidative stress and inflammation in experimental models of acute and chronic lung injury in mice resulting from diverse toxicants. The development of lung tissue damage in response to direct or indirect oxidant stress is a complex process, associated with changes in expression levels of a number of genes. We therefore postulated that flaxseed might modulate gene expression of vital signaling pathways, thus interfering with the development of tissue injury. We evaluated gene expression in lungs of flaxseed-fed (10%FS) mice under unchallenged, control conditions. We reasoned that array technology would provide a powerful tool for studying the mechanisms behind this response and aid the evaluation of dietary flaxseed intervention with a focus on toxicologically relevant molecular gene targets. Gene expression levels in lung tissues were analyzed using a large-scale array whereby 28,800 genes were evaluated. 3,713 genes (12.8%) were significantly (p < 0.05) differentially expressed, of which 2,088 had a >1.5-fold change. Genes affected by FS include those in protective pathways such as Phase I and Phase II. The array studies have provided information on how FS modulates gene expression in lung and how they might be related to protective mechanisms. In addition, our study has confirmed that flaxseed is a nutritional supplement with potentially useful therapeutic applications in complementary and alternative (CAM) medicine especially in relation to treatment of lung disease.


How Parents of Children With Cancer Learn About Their Children's Prognosis.
To determine which prognostic information sources parents find informative and which are associated with better parental understanding of prognosis. Prospective, questionnaire-based cohort study of parents and physicians of children with cancer at 2 academic pediatric hospitals. We asked parents how they learned about prognoses and evaluated relationships between information sources and prognostic understanding, defined as accuracy versus optimism. We excluded parents with pessimistic estimates and whose children had such good prognoses that optimism relative to the physician was impossible. Analytic cohort of 256 parent-physician pairs. Most parents considered explicit sources (conversations with oncologists at diagnosis, day-to-day conversations with oncologists, and conversations with nurses) "very" or "extremely" informative (73%-85%). Implicit sources (parent's sense of how child was doing or how oncologist seemed to feel child was doing) were similarly informative (84%-87%). Twenty-seven percent (70/253) of parents reported prognostic estimates matching physicians' estimates. Parents who valued implicit information had lower prognostic accuracy (odds ratio [OR] 0.50; 95% confidence interval 0.29-0.88), especially those who relied on a "general sense of how my child's oncologist seems to feel my child is doing" (OR 0.47; 0.22-0.99). Parents were more likely to use implicit sources if they reported receiving high-quality prognostic information (OR 3.02; 1.41-6.43), trusted the physician (OR 2.01; 1.01-3.98), and reported high-quality physician communication (OR 1.81; 1.00-3.27). Reliance on implicit sources was associated with overly-optimistic prognostic estimates. Parents who endorsed strong, trusting relationships with physicians were not protected against misinformation.


[Effect of carbon dioxide (CO2) pressure on minute ventilation, parameters of gas exchange and blood gases during hemodialysis using fluid containing acetate and bicarbonate buffers].
In 20 patients with chronic renal failure on a hemodialysis with acetate-containing dialysing fluid gasometric, ventilation and breathing patterns disturbances were determined. The loss of CO2 in the dialysate is attributed the major cause of hypoxemia due to alveolar hypoventilation. Hemodialysis with bicarbonate-containing dialysate can be performed in the absence of any change in ventilation and PaO2 despite a systemic alkalosis. Hyperventilation during HD with high concentration of bicarbonate indicate that changes in CO2 tension in the pulmonary circulation can lead to a change in minute ventilation due to the presence of slowly adapting pulmonary chemoreceptors. In patients with low respiratory response, respiratory muscle weakness intensified additionally by hypercapnia may explain this phenomenon.


Co-Inheritance Analysis within the Domains of Life Substantially Improves Network Inference by Phylogenetic Profiling.
Phylogenetic profiling, a network inference method based on gene inheritance profiles, has been widely used to construct functional gene networks in microbes. However, its utility for network inference in higher eukaryotes has been limited. An improved algorithm with an in-depth understanding of pathway evolution may overcome this limitation. In this study, we investigated the effects of taxonomic structures on co-inheritance analysis using 2,144 reference species in four query species: Escherichia coli, Saccharomyces cerevisiae, Arabidopsis thaliana, and Homo sapiens. We observed three clusters of reference species based on a principal component analysis of the phylogenetic profiles, which correspond to the three domains of life-Archaea, Bacteria, and Eukaryota-suggesting that pathways inherit primarily within specific domains or lower-ranked taxonomic groups during speciation. Hence, the co-inheritance pattern within a taxonomic group may be eroded by confounding inheritance patterns from irrelevant taxonomic groups. We demonstrated that co-inheritance analysis within domains substantially improved network inference not only in microbe species but also in the higher eukaryotes, including humans. Although we observed two sub-domain clusters of reference species within Eukaryota, co-inheritance analysis within these sub-domain taxonomic groups only marginally improved network inference. Therefore, we conclude that co-inheritance analysis within domains is the optimal approach to network inference with the given reference species. The construction of a series of human gene networks with increasing sample sizes of the reference species for each domain revealed that the size of the high-accuracy networks increased as additional reference species genomes were included, suggesting that within-domain co-inheritance analysis will continue to expand human gene networks as genomes of additional species are sequenced. Taken together, we propose that co-inheritance analysis within the domains of life will greatly potentiate the use of the expected onslaught of sequenced genomes in the study of molecular pathways in higher eukaryotes.


What should be included in the assessment of laypersons' paediatric basic life support skills? Results from a Delphi consensus study.
Assessment of laypersons' Paediatric Basic Life Support (PBLS) skills is important to ensure acquisition of effective PBLS competencies. However limited evidence exists on which PBLS skills are essential for laypersons. The same challenges exist with respect to the assessment of foreign body airway obstruction management (FBAOM) skills. We aimed to establish international consensus on how to assess laypersons' PBLS and FBAOM skills. A Delphi consensus survey was conducted. Out of a total of 84 invited experts, 28 agreed to participate. During the first Delphi round experts suggested items to assess laypersons' PBLS and FBAOM skills. In the second round, the suggested items received comments from and were rated by 26 experts (93%) on a 5-point scale (1 = not relevant to 5 = essential). Revised items were anonymously presented in a third round for comments and 23 (82%) experts completed a re-rating. Items with a score above 3 by more than 80% of the experts in the third round were included in an assessment instrument. In the first round, 19 and 15 items were identified to assess PBLS and FBAOM skills, respectively. The ratings and comments from the last two rounds resulted in nine and eight essential assessment items for PBLS and FBAOM skills, respectively. The PBLS items included: "Responsiveness"," Call for help", "Open airway"," Check breathing", "Rescue breaths", "Compressions", "Ventilations", "Time factor" and "Use of AED". The FBAOM items included: "Identify different stages of foreign body airway obstruction", "Identify consciousness", "Call for help", "Back blows", "Chest thrusts/abdominal thrusts according to age", "Identify loss of consciousness and change to CPR", "Assessment of breathing" and "Ventilation". For assessment of laypersons some PBLS and FBAOM skills described in guidelines are more important than others. Four out of nine of PBLS skills focus on airway and breathing skills, supporting the major importance of these skills for laypersons' resuscitation attempts. International consensus on how to assess laypersons' paediatric basic life support and foreign body airway obstruction management skills was established. The assessment of these skills may help to determine when laypersons have acquired competencies. Not relevant.


Increasing Pain Interference Is Associated With Cognitive Decline Over Four Years Among Older Puerto Rican Adults.
Pain is associated with cognitive decline among older adults, but few studies have investigated bidirectional associations between pain and cognitive decline, especially in older Hispanic populations. Our objective was to assess the bidirectional association between pain interference and cognitive performance in a sample of older Puerto Rican adults. Data came from baseline and 4-year follow-up of the Puerto Rican Elderly: Health Conditions Study, a longitudinal representative study of Puerto Rican older adults aged 60 and older. Pain and cognitive performance were assessed at each wave. A pain interference variable was created using the sum of pain status (yes/no) and pain interference (yes/no; range 0-2). Global cognitive performance was assessed with the Mini-Mental Cabán. We tested bidirectional associations using a path model with concurrent and cross-lagged paths between pain and cognitive performance, adjusting for sociodemographic and health factors (n = 2 349). Baseline pain interference was not associated with baseline cognitive performance (p = .636) or with cognitive performance at follow-up (p = .594). However, increased pain interference at follow-up was associated with greater cognitive decline at follow-up (β = -0.07, standard error [SE] = 0.02, p = .003). Greater baseline cognitive performance was associated with lower pain interference at follow-up (β = -0.07, SE = 0.02, p = .007). These findings highlight the importance of worsening pain interference as a potentially modifiable risk factor for cognitive decline, as pain treatment options exist. Additionally, better baseline cognitive performance may be a protective factor for pain, providing further evidence of the dynamic relationship between pain and cognitive performance.


The Prevalence of Inappropriate Image Duplication in Biomedical Research Publications.
Inaccurate data in scientific papers can result from honest error or intentional falsification. This study attempted to determine the percentage of published papers that contain inappropriate image duplication, a specific type of inaccurate data. The images from a total of 20,621 papers published in 40 scientific journals from 1995 to 2014 were visually screened. Overall, 3.8% of published papers contained problematic figures, with at least half exhibiting features suggestive of deliberate manipulation. The prevalence of papers with problematic images has risen markedly during the past decade. Additional papers written by authors of papers with problematic images had an increased likelihood of containing problematic images as well. As this analysis focused only on one type of data, it is likely that the actual prevalence of inaccurate data in the published literature is higher. The marked variation in the frequency of problematic images among journals suggests that journal practices, such as prepublication image screening, influence the quality of the scientific literature.


Lower initial central venous pressure in septic patients from long-term care facilities than in those from the community.
The cornerstone of emergency management of severe sepsis and septic shock is early (within 6 hours) goal-directed therapy, including maintenance of central venous pressure (CVP) at 8-12 mmHg. It is unclear whether there is a difference in initial (baseline) CVP between septic patients who are referred from the community and those who come from long-term care facilities (LTCFs) in Taiwan. We designed this study to investigate the difference in hemodynamic parameters between these two groups. Every patient with severe sepsis or septic shock who had a central venous catheter inserted via the internal jugular or subclavian vein at Kaohsiung Medical University Hospital between April 2007 and October 2007 was enrolled. CVP was measured immediately at the emergency department. Patient demographics, including residence, were retrospectively recorded and analyzed. There were 166 evaluable patients; 125 (75.3%) came from the community and 41 (24.7%) from LTCFs. There were no significant differences in age, sex, initial body temperature, heart rate, blood pressure, or leukocyte count between the two groups. However, patients who were referred from LTCFs had a significantly lower initial CVP than those from the community (5.0 ± 4.5 mmHg vs. 7.0 ± 4.8 mmHg, p = 0.023). The difference was more significant between mechanically ventilated patients from LTCFs and those from the community (5.0 ± 3.0 mmHg vs. 8.1 ± 5.6 mmHg, p = 0.006). Severely septic patients referred from LTCFs may require more aggressive fluid resuscitation within the first 6 hours of the diagnostic criteria met at the emergency department to achieve the CVP target of early goal-directed therapy.


The synergistic upregulation of phase II detoxification enzymes by glucosinolate breakdown products in cruciferous vegetables.
Cruciferous vegetables contain secondary metabolites termed glucosinolates that break down to products that upregulate hepatic detoxification enzymes. We have previously shown that a mixture of four major glucosinolate breakdown products from Brussels sprouts interact to produce synergistic induction of phase II detoxification enzymes. Here we tested the hypothesis that this synergism is at the level of transcription and is due to the interaction between the oral bifunctional inducer, indole-3-carbinol (I3C), and monofunctional inducer, crambene (1-cyano 2-hydroxy 3-butene). Adult male rats were treated by gavage with either corn oil (vehicle); crambene (50 mg/kg), I3C (56 mg/kg), or a mix of crambene and I3C at the doses shown. Given orally, I3C alone and crambene with I3C caused significant induction of CYP1A activity and CYP1A1 mRNA levels, whereas crambene alone had no significant effect on CYP1A activity or mRNA levels. Crambene and I3C individually caused induction of glutathione S-transferase (GST) and quinone reductase (QR) activity. The mixture of crambene and I3C caused induction of GST and QR that was significantly greater than the sum of the induction by individual treatments. Upregulation of total GST activity was not as great as that of QR, possibly because some subunits did not show this effect. GST Ya2 mRNA showed a synergistic upregulation by crambene and I3C, while Yc1 and Yc2 showed only an additive response. We speculate that this different regulation is partly due to differences in gene sequences within the antioxidant response element and xenobiotic response element in the regulatory region of GST Ya2 compared to those within the regulatory region of the Yc1/Yc2 subunits.


Inhibition of serum- and ras-stimulated DNA synthesis by antibodies to phospholipase C.
Several immunologically distinct isozymes of inositol phospholipid-specific phospholipase C (PLC) have been purified from bovine brain. Murine NIH 3T3 fibroblasts were found to express PLC-gamma, but the expression of PLC-beta was barely detectable by radioimmunoassay or protein immunoblot. A mixture of monoclonal antibodies was identified that neutralizes the biological activity of both endogenous and injected purified PLC-gamma. When co-injected with oncogenic Ras protein or PLC-gamma, this mixture of antibodies inhibited the induction of DNA synthesis that characteristically results from the injection of these proteins into quiescent 3T3 cells. However, when oncogenic Ras protein or PLC-gamma was co-injected with a neutralizing monoclonal antibody to Ras, only the DNA synthesis induced by the Ras protein was inhibited--that induced by PLC was unaffected. These results suggest that the Ras protein is an upstream effector of PLC activity in phosphoinositide-specific signal transduction and that PLC-gamma activity is necessary for Ras-mediated induction of DNA synthesis.


Adjuvant chemotherapy in early breast cancer.
Despite living in an era of progressively improved molecular characterization of breast cancer with novel prognostic and predictive tests as well as increased use of targeted therapies, adjuvant chemotherapy is still a cornerstone in the treatment of early breast cancer. Numerous clinical trials of adjuvant chemotherapy without trastuzumab have clearly shown that the effectiveness depends not only on the mere application of new substances (e.g., taxanes) but at least equally important on the way to utilize them. At present, standard adjuvant chemotherapy should include anthracyclines, taxanes and cyclophosphamide. Docetaxel is best used in three weekly intervals, while paclitaxel should be delivered either weekly or dose-dense every two weeks with G-CSF support. In high risk breast cancer patients with more than three involved axillary lymph nodes, an intensified dose-dense and sequential approach leads to significantly improved survival. Other approaches to improve the efficacy of adjuvant chemotherapy employ the incorporation of additional substances like capecitabine. Conversely, investigators trying to de-escalate adjuvant chemotherapy implemented taxane-containing but anthracycline-free chemotherapy. Altogether, these new approaches are awaiting further confirmatory clinical trials before they should be regarded as standard of care in early breast cancer.


An evaluation of the challenges to developing tumor BRCA1 and BRCA2 testing methodologies for clinical practice.
Ovarian cancer patients with germline or somatic pathogenic variants benefit from treatment with poly ADP ribose polymerase (PARP) inhibitors. Tumor BRCA1/2 testing is more challenging than germline testing as the majority of samples are formalin-fixed paraffin embedded (FFPE), the tumor genome is complex, and the allelic fraction of somatic variants can be low. We collaborated with 10 laboratories testing BRCA1/2 in tumors to compare different approaches to identify clinically important variants within FFPE tumor DNA samples. This was not a proficiency study but an inter-laboratory comparison to identify common issues. Each laboratory received the same tumor DNA samples ranging in genotype, quantity, quality, and variant allele frequency (VAF). Each laboratory performed their preferred next-generation sequencing method to report on the variants. No false positive results were reported in this small study and the majority of methods detected the low VAF variants. A number of variants were not detected due to the bioinformatics analysis, variant classification, or insufficient DNA. The use of hybridization capture or short amplicon methods are recommended based on a bioinformatic assessment of the data. The study highlights the importance of establishing standards and standardization for tBRCA testing particularly when the test results dictate clinical decisions regarding life extending therapies.


Test of a two-pathway model for small-solute exchange across the capillary wall.
We previously proposed a two-pathway model for solute and water transport across vascular endothelium (Fu, B. M., R. Tsay, F. E. Curry, and S. Weinbaum. J. Biomech. Eng. 116: 502-513, 1994) that hypothesized the existence of a continuous slit 2 nm wide along tight junction strands within the interendothelial cleft in parallel with 20 x 150-nm breaks in tight junctions. We tested this model by measuring capillary permeability coefficients (P) to a small solute (sodium fluorescein, radius 0.45 nm), assumed to permeate primarily the 2-nm small pore, and an intermediate-sized solute (FITC-alpha-lactalbumin, radius 2.01 nm) excluded from the small pore. Mean values of the paired diffusive permeability coefficients, Psodium fluorescein and PFITC-alpha-lactalbumin, were 34.4 and 2.9 x 10(-6) cm/s, respectively, after corrections for solvent drag and free dye (n = 26). These permeabilities were accounted for by transport through the large-break pathway without the additional capacity of the hypothetical 2-nm pathway. As a further test we examined the relative reductions of Psodium fluorescein and PFITC-alpha-lactalbumin produced by elevated intracellular cAMP. Within 20 min after the introduction of rolipram and forskolin, Psodium fluorescein and PFITC-alpha-lactalbumin decreased to 0.67 and 0.64 times their respective baseline values. These similar responses to permeability decrease were evidence that the two solutes were carried by a common pathway. Combined results in both control and reduced permeability states did not support the hypothesis that a separate pathway across tight junctions is available for solutes with a radius as large as 0.75 nm. If such a pathway is present, then its size must be smaller than that of sodium fluorescein.


Structural, electronic, and linear optical properties of organic photovoltaic PBTTT-C14 crystal.
Poly(2,5-bis(3-tetradecylthiophen-2yl)thieno(3,2-b)thiophene) (PBTTT-C14) is an important electro-optical polymer, whose three-dimensional crystal structure is somewhat ambiguous and the fundamental electronic and linear optical properties are not well known. We carried out first-principles calculations to model the crystal structure and to study the effect of side-chains on the physical structure and electronic properties. Our calculations suggest that the patterns of side-chain has little direct effect on the valence band maximum and conduction band minimum but they do have impact on the bandgap through changing the π-π stacking distance. By examining the band structure and wave functions, we conclude that the fundamental bandgap of the PBTTT-C14 crystal is determined by the conduction band energy at the Q point. The calculations indicate that the bandgap of PBTTT-C14 crystal may be tunable by introducing different side-chains. The significant peak in the imaginary part of the dielectric function arises from transitions along the polymer backbone axis, as determined by the critical-point analysis and the large optical transition matrix elements in the direction of the backbone.


Seasonal variation in heat shock proteins Hsp70 and Hsp90 expression in an exposed and a shaded habitat of Iris pumila.
Seasonal variation in heat shock proteins Hsp70 and Hsp90 expression was studied in the leaves of two naturally growing Iris pumila populations, one inhabiting an open dune site, and the other the understorey of a Pinus silvestris stand. The Hsps were quantified by an immunoblotting procedure. The level of the Hsps was found to vary significantly both across seasons and between habitats. The mean Hsp70 concentration was significantly greater at the open area than in the woodland understorey, reaching its maximum in the summer, especially in plants experiencing full sunlight. Two Hsp90 isoforms, referred to as Hsp90a (86 kDa) and Hsp90b (84 kDa), were detected. At both habitats, the level of Hsp90a was highest in autumn, that of Hsp90b in spring, whereas both of them reached a nadir in summer. Throughout the growing season, the relative abundance of Hsp90b was higher in plants growing under vegetation canopy in comparison to those inhabiting the open dune site. An inverse relationship between the phenotypic variation in specific leaf area and the level of Hsp90b over seasons at both habitats was observed, suggesting the role of this protein in buffering phenotypic variation in the wild.


The CryoEM structure of the Saccharomyces cerevisiae ribosome maturation factor Rea1.
The biogenesis of 60S ribosomal subunits is initiated in the nucleus where rRNAs and proteins form pre-60S particles. These pre-60S particles mature by transiently interacting with various assembly factors. The ~5000 amino-acid AAA+ ATPase Rea1 (or Midasin) generates force to mechanically remove assembly factors from pre-60S particles, which promotes their export to the cytosol. Here we present three Rea1 cryoEM structures. We visualise the Rea1 engine, a hexameric ring of AAA+ domains, and identify an α-helical bundle of AAA2 as a major ATPase activity regulator. The α-helical bundle interferes with nucleotide-induced conformational changes that create a docking site for the substrate binding MIDAS domain on the AAA +ring. Furthermore, we reveal the architecture of the Rea1 linker, which is involved in force generation and extends from the AAA+ ring. The data presented here provide insights into the mechanism of one of the most complex ribosome maturation factors.


Exome sequencing identifies putative drivers of progression of transient myeloproliferative disorder to AMKL in infants with Down syndrome.
Some neonates with Down syndrome (DS) are diagnosed with self-regressing transient myeloproliferative disorder (TMD), and 20% to 30% of those progress to acute megakaryoblastic leukemia (AMKL). We performed exome sequencing in 7 TMD/AMKL cases and copy-number analysis in these and 10 additional cases. All TMD/AMKL samples contained GATA1 mutations. No exome-sequenced TMD/AMKL sample had other recurrently mutated genes. However, 2 of 5 TMD cases, and all AMKL cases, showed mutations/deletions other than GATA1, in genes proven as transformation drivers in non-DS leukemia (EZH2, APC, FLT3, JAK1, PARK2-PACRG, EXT1, DLEC1, and SMC3). One patient at the TMD stage revealed 2 clonal expansions with different GATA1 mutations, of which 1 clone had an additional driver mutation. Interestingly, it was the other clone that gave rise to AMKL after accumulating mutations in 7 other genes. Data suggest that GATA1 mutations alone are sufficient for clonal expansions, and additional driver mutations at the TMD stage do not necessarily predict AMKL progression. Later in infancy, leukemic progression requires "third-hit driver" mutations/somatic copy-number alterations found in non-DS leukemias. Putative driver mutations affecting WNT (wingless-related integration site), JAK-STAT (Janus kinase/signal transducer and activator of transcription), or MAPK/PI3K (mitogen-activated kinase/phosphatidylinositol-3 kinase) pathways were found in all cases, aberrant activation of which converges on overexpression of MYC.


Parallels and divergences in the acquisition and dissolution of language.
Analyses of the relation between the acquisition and dissolution of language have typically focused on whether or not the forms of language dysfunction that result from brain damage correspond to specific stages of language acquisition: the Regression Hypothesis. These analyses have not met with great success: although there are a number of superficial similarities between aphasic disorders and different stages of the immature linguistic system, there are also important differences. I will suggest that a focus on the behavioral similarities between language acquisition and dissolution is unlikely to be productive. A more productive course would be to focus, instead, on the general principles that constrain the acquisition and dissolution of language.


A new avian fibroblast growth factor receptor in myogenic and chondrogenic cell differentiation.
We studied the expression of FREK (fibroblast growth factor receptor-like embryonic kinase), a new receptor recently cloned from quail embryo, during the differentiation of skeletal muscle satellite cells and epiphyseal growth-plate chondrocytes. Although FREK mRNA was expressed in both cell types, satellite cells expressed higher levels of this mRNA than chondrocytes. FREK gene expression was found to be modulated by b-FGF in a biphasic manner: low concentrations increased expression, whereas high concentrations attenuated it. In both cell cultures, the levels of FREK mRNA declined during terminal differentiation. Moreover, retinoic acid (RA), which induces skeletal muscle satellite cells to differentiate, also caused a reduction in FREK gene expression in these cells. Induction of chondrocyte differentiation with ascorbic acid was monitored by a decrease in collagen type II gene expression and an increase in alkaline phosphatase activity. Satellite cell differentiation was marked by morphological changes as well as by increased sarcomeric myogenin content and creatine kinase activity and changes in the expression of the regulatory muscle-specific genes, MyoD and myogenin. DNA synthesis in both cell types was stimulated by b-FGF. However, in satellite cells, the response was bell-shaped, peaking at 1 ng/ml b-FGF, whereas in chondrocytes, higher levels of b-FGF were needed. b-FGF-dependent DNA synthesis in satellite cells was decreased by RA at concentrations over 10(-7) M. The observed correlation between the level of FREK gene expression and various stages of differentiation, its modulation by b-FGF and RA, as well as the correlation between FREK gene expression and the physiological response to b-FGF, suggest that this specific FGF receptor plays an important role in muscle and cartilage cell differentiation.


The orphan mouse receptor interleukin (IL)-8R beta binds N51. Structure-function analysis using N51/IL-8 chimeric molecules.
We have demonstrated that the orphan receptor representing the putative mouse (mu) homolog of the human (hu) interleukin-8 receptor beta (IL-8R beta) binds the mouse N51 cytokine, also known as KC. The muIL-8R beta gene was constitutively expressed in NIH 3T3 cells (NIH-muIL-8R beta). Cells and plasma membranes from the NIH-muIL-8R beta clone showed binding of 125I-N51 that was displaced by unlabeled N51. Other related cytokines were assayed for their ability to displace 125I-N51. MIP-2 and GRO alpha/MGSA competed as well as N51 for the receptor, but huIL-8 and NAP-2 did not compete at all. Chimeric molecules between IL-8 and N51 were used to extend the binding analysis. The segment between the conserved cysteines 2 and 3, named domain I; cysteines 3 and 4, domain II; and cysteine 4 and the C terminus, domain III of IL-8 were replaced by the corresponding domains of N51 and vice versa. When studying the binding of 125I-N51 and the hybrid molecules to the receptor, we observed that chimeras of N51 containing either domain I, II, or III of IL-8 were agonists of N51, and chimeras of IL-8 containing domain II or III of N51 were partial agonists of N51. These results demonstrate that domain I of N51 does not confer binding specificity and suggest that the region from the third cysteine to the C terminus of the N51 molecule is more important for binding to muIL-8R beta.


Effect of phenylalanine metabolites on the activities of enzymes of ketone-body utilization in brain of suckling rats.
1. The effects of phenylalanine and its metabolites (phenylacetate, phenethylamine, phenyl-lactate, o-hydroxyphenylacetate and phenylpyruvate) on the activity of 3-hydroxybutyrate dehydrogenase (EC 220.127.116.11) 3-oxo acid CoA-transferase (EC 18.104.22.168) and acetoacetyl-CoA thiolase (EC 22.214.171.124) in brain of suckling rats were investigated. 2. The 3-hydroxybutyrate dehydrogenase from the brain of suckling rats had a Km for 3-hydroxybutyrate of 1.2 mM. Phenylpyruvate, phenylacetate and o-hydroxyphenylacetate inhibited the enzyme activity with Ki values of 0.5, 1.3 and 4.7 mM respectively. 3. The suckling-rat brain 3-oxo acid CoA-transferase activity had a Km for acetoacetate of 0.665 mM and for succinyl (3-carboxypropionyl)-CoA of 0.038 mM. The enzyme was inhibited with respect to acetoacetate by phenylpyruvate (Ki equals 1.3 mM) and o-hydroxyphenylacetate (Ki equals 4.5 mM). The reaction in the direction of acetoacetate was also inhibited by phenylpyruvate (Ki equals 1.6 mM) and o-hydroxyphenylacetate (Ki equals 4.5 mM). 4. Phenylpyruvate inhibited with respect to acetoacetyl-CoA both the mitochondrial (Ki equals 3.2 mM) and cytoplasmic (Ki equals 5.2 mM) acetoacetyl-CoA thiolase activities. 5. The results suggest that inhibition of 3-hydroxybutyrate dehydrogenase and 3-oxo acid CoA-transferase activities may impair ketone-body utilization and hence lipid synthesis in the developing brain. This suggestion is discussed with reference to the pathogenesis of mental retardation in phenylketonuria.


RyR-mediated Ca2+ release elicited by neuronal activity induces nuclear Ca2+ signals, CREB phosphorylation, and Npas4/RyR2 expression.
The expression of several hippocampal genes implicated in learning and memory processes requires that Ca2+ signals generated in dendritic spines, dendrites, or the soma in response to neuronal stimulation reach the nucleus. The diffusion of Ca2+ in the cytoplasm is highly restricted, so neurons must use other mechanisms to propagate Ca2+ signals to the nucleus. Here, we present evidence showing that Ca2+ release mediated by the ryanodine receptor (RyR) channel type-2 isoform (RyR2) contributes to the generation of nuclear Ca2+ signals induced by gabazine (GBZ) addition, glutamate uncaging in the dendrites, or high-frequency field stimulation of primary hippocampal neurons. Additionally, GBZ treatment significantly increased cyclic adenosine monophosphate response element binding protein (CREB) phosphorylation-a key event in synaptic plasticity and hippocampal memory-and enhanced the expression of Neuronal Per Arnt Sim domain protein 4 (Npas4) and RyR2, two central regulators of these processes. Suppression of RyR-mediated Ca2+ release with ryanodine significantly reduced the increase in CREB phosphorylation and the enhanced Npas4 and RyR2 expression induced by GBZ. We propose that RyR-mediated Ca2+ release induced by neuronal activity, through its contribution to the sequential generation of nuclear Ca2+ signals, CREB phosphorylation, Npas4, and RyR2 up-regulation, plays a central role in hippocampal synaptic plasticity and memory processes.


Characterization of antigen-specific, Ia-restricted, L3T4+ cytolytic T lymphocytes and assessment of thymic influence on their self specificity.
The goals of the present study were: (a) to generate antigen-specific L3T4+ cytolytic T lymphocytes (CTL), (b) to determine their major histocompatibility complex (MHC) restriction specificity, and (c) to assess the influence of thymic MHC determinants on their self specificity. We found that L3T4+ CTL specific for either trinitrophenyl (TNP)-modified self determinants or minor histocompatibility antigens could be generated from Lyt-2- responder T cells provided that the response cultures were supplemented with supernatants rich in helper factors. Such antigen-specific L3T4+ CTL were Ia-restricted by the criteria that they lysed only Ia+ target cells and that their lysis of Ia+ target cells was specifically inhibited by anti-Ia monoclonal antibodies. The relative frequency of L3T4+ pCTL was found to be only 5-10% of the total anti-TNP pCTL present in the spleens of normal mice. Finally, we utilized radiation bone marrow chimeras to assess the influence of the thymic haplotype on the self-Ia specificity of L3T4+ CTL. Both bulk culture and limiting dilution experiments revealed that the self-Ia specificity of L3T4+ anti-TNP CTL from F1----parent and A----B allogeneic chimeras was not markedly skewed toward the haplotype of the chimeric thymus. These results contrast with those obtained previously for L3T4+ anti-TNP Th cells and demonstrate that in the radiation bone marrow chimera model of T cell differentiation, the self specificity of Th cells but not pCTL is markedly influenced by the haplotype of the chimeric thymus.


[Squirrel visual cortex neurons selective for contour orientation].
The receptive field organization of orientation-selective neurons was studied in the squirrel visual cortex. Neurons with mutual inhibiting on- and off-areas of the receptive field, partially and completely overlapping excitatory and inhibitory mechanisms were observed. Neurons of the second group are the most typical. They reveal orientation selectivity if the stimuli are in the excitatory area of the receptive field, the inhibitory areas outside the excitatory area sharpen the selectivity. It is supposed that no obvious differentiation between simple and complex neurons exist in the squirrel visual cortex.


Prospective study of bacterial meningitis in North East Thames region, 1991-3, during introduction of Haemophilus influenzae vaccine.
To describe the epidemiology of primary bacterial meningitis in the North East Thames region over a three year period before and during the introduction of the vaccine for Haemophilus influenzae type b. Analysis of information on cases of primary bacterial meningitis identified by microbiology laboratories in the region, with collection of case data by questionnaire. Annual incidence rates for types of meningitis according to age and ethnic group. The annual incidence rates for the three major causes of bacterial meningitis in the general population were 1.9/100,000 for Neisseria meningitidis, 1.6/100,000 for Haemophilus influenzae before vaccination, and 1.0/100,000 for Streptococcus pneumoniae. Higher rates of H influenzae meningitis were found in Asians compared with white people (3.6/100,000 v 1.5/100,000, P = 0.01). As a result of the vaccine programme introduced in October 1992 the number of cases of H influenzae meningitis in children under 5 years has fallen by 87%. Bacterial meningitis is a serious problem especially in preschool children. There are differences in the incidence of some causes of bacterial meningitis in different ethnic groups; with H influenzae type b being significantly more common among black and Asian people than among white people. The immunisation programme for H influenzae type b in the North East Thames region has been successful in reducing the incidence of this type of meningitis in Asian and white populations. The numbers were too small to evaluate in the black population.


Training community therapists to deliver cognitive behavioral therapy in the aftermath of disaster.
In this paper we examine the effectiveness of disseminating Cognitive Behavioral Therapy for Postdisaster Distress (CBT-PD) to community therapists in Baton Rouge, Louisiana following Hurricane Katrina. One hundred four therapists attended a two-day training in CBT-PD with on-going case consultation. Pre- and post-training, therapists rated eight core cognitive behavioral therapy elements on their importance, how well they understood how to deliver the element, and how confident they were in their ability to deliver the element. Post-training they completed a CBT-PD knowledge questionnaire and session fidelity forms. Seventy-seven clients completed satisfaction questionnaires and reported on how often they utilized the skills taught in CBT-PD. Therapists showed significant improvements in their ratings of the importance of various elements of cognitive behavioral therapy, their knowledge and understanding of those elements, and their confidence that they could use them effectively. Immediately following the training 90% of therapists demonstrated excellent retention of CBT-PD. Self-report measures from both therapists and clients indicated that critical session elements were delivered. This work suggests that CBT-PD can be applied in a real-world setting and that community therapists can be trained in relatively short time spans with on-going support. This finding is especially important in the disaster field given that communities are likely to find themselves in emergency situations in which a number of non-expert trauma therapists will need to deliver trauma services.


[Listeria in plants: an experimental study of its colonization, numbers and variability].
The possibility for Listeria monocytogenes to penetrate into plants from the soil via the root system was experimentally proved. Listeria were shown to continuously persist for 30 days (the term of observation) in the vegetative organs of wheat (roots, stems and leaves). The concentration of Listeria was 10(9) CFU/g in the environment (soil extract) and the roots of wheat, 10(6)-10(7) CFU/g in stems, 10(8)-10(9) CFU/g in leaves. Six days later the dissociation of colonies in the S-form into small (up to 1 mm) and large (3-4 mm) was observed; in contrast to Listeria in large colonies, those in small colonies had high catalase activity and pronounced cytopathogenic action. The problem of the possible role of plants as the natural reservoir of bacteria, pathogenic for humans and animals, is discussed.


Evaluation of dietary quality of adolescents using Healthy Eating Index.
The aim of this study was to use the Healthy Eating Index-2005 (HEI-2005) to assess diet quality and determine the relationship between the HEI-2005 and the energy and nutrient intakes of adolescents. A cross-sectional study was conducted on 1,104 healthy adolescents, mean age of 15.8 ± 1.24 years. Dietary intake was measured with the 24-hour dietary recall method, and dietary quality was assessed by means of the HEI-2005. Diet quality scores ranged from 23.7 to 77.5. The mean score was found to be 51.5 ± 9.07 according to the HEI-2005. There were no differences according to gender; 42.8% had a poor diet and 57.2% had a diet that needs improvement. No subjects had a "good diet". Lower mean subgroup scores were found for whole grains, total vegetables, total fruits, dairy products, and meat and beans. Fruits and vegetables scores were significantly high in girls, whereas sodium, oil, and meat and beans scores were significantly high in boys. Total HEI-2005 scores were increasingly associated with parental education level when age and gender were adjusted. There was a negative correlation between HEI-2005 scores and age, total energy intake, and fat intake. Positive correlation was only observed in the HEI-2005 scores for protein and dietary fibre intakes. Consequently, the overall diet quality and nutritional habits of Turkish adolescents need modification and improvement. In the family, measures should be initiated by the government, including advertisements and campaigns.


The Cannabinoid Receptor 1 Reverse Agonist AM251 Ameliorates Radiation-Induced Cognitive Decrements.
Despite advancements in the radiotherapeutic management of brain malignancies, resultant sequelae include persistent cognitive dysfunction in the majority of survivors. Defining the precise causes of normal tissue toxicity has proven challenging, but the use of preclinical rodent models has suggested that reductions in neurogenesis and microvascular integrity, impaired synaptic plasticity, increased inflammation, and alterations in neuronal structure are contributory if not causal. As such, strategies to reverse these persistent radiotherapy-induced neurological disorders represent an unmet medical need. AM251, a cannabinoid receptor 1 reverse agonist known to facilitate adult neurogenesis and synaptic plasticity, may help to ameliorate radiation-induced CNS impairments. To test this hypothesis, three treatment paradigms were used to evaluate the efficacy of AM251 to ameliorate radiation-induced learning and memory deficits along with disruptions in mood at 4 and 12 weeks postirradiation. Results demonstrated that acute (four weekly injections) and chronic (16 weekly injections) AM251 treatments (1 mg/kg) effectively alleviated cognitive and mood dysfunction in cranially irradiated mice. The beneficial effects of AM251 were exemplified by improved hippocampal- and cortical-dependent memory function on the novel object recognition and object in place tasks, while similar benefits on mood were shown by reductions in depressive- and anxiety-like behaviors on the forced swim test and elevated plus maze. The foregoing neurocognitive benefits were associated with significant increases in newly born (doublecortin+) neurons (1.7-fold), hippocampal neurogenesis (BrdU+/NeuN+mature neurons, 2.5-fold), and reduced expression of the inflammatory mediator HMGB (1.2-fold) in the hippocampus of irradiated mice. Collectively, these findings indicate that AM251 ameliorates the effects of clinically relevant cranial irradiation where overall neurological benefits in memory and mood coincided with increased hippocampal cell proliferation, neurogenesis, and reduced expression of proinflammatory markers.


Double-Negative T Cells Regulate Hepatic Stellate Cell Activation to Promote Liver Fibrosis Progression via NLRP3.
We mainly explored the role and mechanism of double-negative T cells (DNTs) in liver fibrosis. DNTs were co-cultured with mouse hepatic stellate cells (HSCs). Later, cell viability was detected by Cell Counting Kit-8 (CCK-8) assay; α-SMA expression was measured through fluorescence staining; TNF-α, IL-6, and MMP-9 levels were measured by ELISA; and the expression of Bcl-2, TGF-β1, NLRP3, ASC, and TNFR1 proteins in HSCs was detected by Western blotting (WB) assay. At the same time, HSC-NLRP3-/- and HSC-TNFR1-/- are used to explore the mechanism. In mouse experiments, mice were intraperitoneally injected with DNTs; afterward, the hepatic tissue fibrosis degree was detected by Masson staining, α-SMA expression was measured through immunohistochemistry (IHC) assay, and histopathological changes were detected by sirius-red staining and H&E staining. The results suggested that DNTs promoted HSC activation and NLRP3 activation. The effect of DNTs on activating HSC-NLRP3-/- was suppressed, and the difference was significant as compared with HSCs. HSC-TNFR1-/- activation was also inhibited. To explore the mechanism of DNT-secreted TNF-α in TNFR1-NLRP3 activation, we transfected DNTs with TNF-α siRNA; as a result, DNTs with TNF-α silencing did not significantly affect HSC activation. DNTs promoted hepatic tissue fibrosis progression and HSC activation; after treatment with NLRP3 inhibitor, the effect of DNTs on promoting fibrosis was suppressed. We discovered that DNTs played an important role in liver fibrosis and that DNTs promoted HSC activation via the TNF-α-TNFR1-NLRP3 signal axis, thus further promoting liver fibrosis progression.


Seat belt use before and after motor vehicle trauma.
Motor vehicle crashes cause significant morbidity and mortality annually. Seat belt use has partially been associated with a decreased risk of morbidity and mortality among those involved in motor vehicle crashes. Persons injured in motor vehicle crashes and not wearing seat belts have an increased risk of admission to trauma centers for motor vehicle crash-related injury. The purpose of this study was to measure changes in seat belt use after discharge among patients admitted to a Level I trauma center for injuries sustained in motor vehicle crashes. Patients admitted to a Level I trauma center for injuries sustained in motor vehicle crashes during 1998 were eligible for participation. A telephone interview was conducted with a random sample of 136 eligible patients regarding patterns of seat belt use before and after their collision. Demographic data and clinical characteristics were also collected. The frequency of seat belt use before and after crash involvement was compared for all patients and stratified by age, gender, race, and Injury Severity Score (ISS). Slightly over half (54%) of patients reported "always" wearing a seat belt before their collision compared with 85% afterward. Younger age groups, male subjects, and whites had the largest increases in the frequency of seat belt use after collision (45%, 37%, and 44% increases, respectively). With respect to injury severity, the largest increase in the frequency of seat belt use was among those with ISS of 15 to 25 (82% increase). Significant concordance between patient- and emergency medical service-reported use of seat belts was observed. Among subjects reported by emergency medical service personnel to have been restrained, nearly 90% reported belt use at the time of the telephone interview. The most frequently cited occasion for failure to use seat belts (30%) was when taking short trips. Other reported reasons were forgetting to fasten belts (29%), discomfort (10%), being in a rush (8%), riding in the back seat (4%), and that seat belts were unnecessary when riding with a good driver (3%). Involvement in a motor vehicle crash results in increased seat belt use. Prevention efforts should be directed toward those patients who report infrequent use. Patient "converts" to seat belt use after collisions may be useful in public awareness and prevention campaigns.


Defective mitochondrial morphology and bioenergetic function in mice lacking the transcription factor Yin Yang 1 in skeletal muscle.
The formation, distribution, and maintenance of functional mitochondria are achieved through dynamic processes that depend strictly on the transcription of nuclear genes encoding mitochondrial proteins. A large number of these mitochondrial genes contain binding sites for the transcription factor Yin Yang 1 (YY1) in their proximal promoters, but the physiological relevance is unknown. We report here that skeletal-muscle-specific YY1 knockout (YY1mKO) mice have severely defective mitochondrial morphology and oxidative function associated with exercise intolerance, signs of mitochondrial myopathy, and short stature. Gene set enrichment analysis (GSEA) revealed that the top pathways downregulated in YY1mKO mice were assigned to key metabolic and regulatory mitochondrial genes. This analysis was consistent with a profound decrease in the level of mitochondrial proteins and oxidative phosphorylation (OXPHOS) bioenergetic function in these mice. In contrast to the finding for wild-type mice, inactivation of the mammalian target of rapamycin (mTOR) did not suppress mitochondrial genes in YY1mKO mice. Mechanistically, mTOR-dependent phosphorylation of YY1 resulted in a strong interaction between YY1 and the transcriptional coactivator peroxisome proliferator-activated receptor gamma coactivator 1α (PGC1α), a major regulator of mitochondrial function. These results underscore the important role of YY1 in the maintenance of mitochondrial function and explain how its inactivation might contribute to exercise intolerance and mitochondrial myopathies.


Vicianin hydrolase is a novel cyanogenic beta-glycosidase specific to beta-vicianoside (6-O-alpha-L-arabinopyranosyl-beta-D-glucopyranoside) in seeds of Vicia angustifolia.
The cyanogenic disaccharide glycoside, vicianin [mandelonitrile beta-vicianoside (6-O-alpha-L-arabinopyranosyl-beta-D-glucopyranoside)], is accumulated in seeds of Vicia angustifolia var. segetalis. Vicianin hydrolase (VH) catalyzes the hydrolysis of vicianin into mandelonitrile and a disaccharide vicianose. VH was purified from the seeds using DEAE-, CM- and Con A-Sepharose chromatography, and the molecular mass of the purified VH was estimated to be 56 kDa on SDS-PAGE. The N-terminal amino acid sequence of the purified VH was determined, and a cDNA encoding VH was obtained. The deduced VH protein consists of a 509 amino acid polypeptide containing a putative secretion signal peptide. It shares about 50% identity with various kinds of plant beta-glycosidases including tea leaf beta-primeverosidase and furcatin hydrolase, and is classified in family 1 of the glycosyl hydrolases. The VH transcript was detected abundantly in seeds and moderately in flowers, but only slightly in leaves, stems and roots, indicating that the organ distribution of VH expression is similar to that of the substrate vicianin. The recombinant VH was produced in insect cells with a baculovirus system, and was compared with the native VH in terms of substrate specificity. Both enzymes hydrolyzed vicianin to release vicianose, demonstrating that VH is a disaccharide-specific beta-glycosidase. VH also hydrolyzed the mandelonitrile beta-glucoside prunasin to some extent but did not hydrolyze the gentiobioside amygdalin, both of which contain the same aglycone as vicianin. Thus, VH is a unique cyanogenic glycosidase showing high glycone specificity for the disaccharide vicianoside.


Traveller gypsies and primary care.
Traveller gypsies have resided in the British Isles for over 500 years, making them one of our oldest ethnic minorities. They experience widespread prejudice and discrimination from the settled population. In the sphere of health care the marginalization of traveller gypsies has resulted in poor access to services and relative neglect of their health needs. In this paper the health of traveller gypsies is reviewed from the perspective of primary care, and the role of general practitioners in improving health care for this community is discussed.


An analysis of disenrollment from Medicare managed care plans by Medicare beneficiaries with diabetes.
The purpose of this work is to determine whether high-cost high-risk Medicare patients with diabetes in managed care plans disenroll more quickly than lower-cost lower-risk Medicare patients with diabetes. If high-cost high-risk patients with diabetes do disenroll more quickly, Medicare managed care plans benefit financially from favorable disenrollment. Time in a health maintenance organization (HMO) was modeled using a duration model with the number of months in the HMO as the dependent variable, controlling for censoring. Data were drawn from a representative sample of Medicare patients with diabetes in the FFS sector in 1994. The panel was followed for 4 years, 1995-1998. The sample included all 6839 individuals who enrolled in a Medicare HMO for at least 1 month during the 48-month observation window. We found a statistically significant negative association between the time in an HMO and pre-enrollment Part B expenditures (beta = -0.00001, t = -4.39) and any Part A expenditures (beta = -0.465, t = -1.98), and 2 of 4 diabetic complications (heart complications: beta = -0.0773, t = -4.61; vision complications beta = -0.2474, t = -1.94). Of the plan characteristics, only the drug benefit variable (beta = 0.151, t = 5.64) had a statistically significant coefficient. Overall, our results support the hypothesis that high-cost, high-risk individuals disenroll from Medicare HMOs sooner than lower-cost lower-risk individuals. However, this effect is mitigated by plans offering better prescription drug benefits. We did find some evidence that patients with diabetes with very high pre-enrollment Part A costs may remain longer in HMOs relative to patients with diabetes with lower Part A prior year expenditures.


Genomic surveillance reveals dynamic shifts in the connectivity of COVID-19 epidemics.
The maturation of genomic surveillance in the past decade has enabled tracking of the emergence and spread of epidemics at an unprecedented level. During the COVID-19 pandemic, for example, genomic data revealed that local epidemics varied considerably in the frequency of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) lineage importation and persistence, likely due to a combination of COVID-19 restrictions and changing connectivity. Here, we show that local COVID-19 epidemics are driven by regional transmission, including across international boundaries, but can become increasingly connected to distant locations following the relaxation of public health interventions. By integrating genomic, mobility, and epidemiological data, we find abundant transmission occurring between both adjacent and distant locations, supported by dynamic mobility patterns. We find that changing connectivity significantly influences local COVID-19 incidence. Our findings demonstrate a complex meaning of "local" when investigating connected epidemics and emphasize the importance of collaborative interventions for pandemic prevention and mitigation.


The Role of Angiotensin II and Cyclic AMP in Alveolar Active Sodium Transport.
Active alveolar fluid clearance is important in keeping airspaces free of edema. Angiotensin II plays a role in the pathogenesis of hypertension, heart failure and others. However, little is known about its contribution to alveolar fluid clearance. Angiotensin II effects are mediated by two specific receptors; AT1 and AT2. The localization of these two receptors in the lung, specifically in alveolar epithelial cells type II, was recently reported. We hypothesize that Angiotensin II may have a role in the regulation of alveolar fluid clearance. We investigated the effect of Angiotensin II on alveolar fluid clearance in rats using the isolated perfused lung model and isolated rat alveolar epithelial cells. The rate of alveolar fluid clearance in control rats was 8.6% ± 0.1 clearance of the initial volume and decreased by 22.5%, 28.6%, 41.6%, 48.7% and 39% in rats treated with 10-10 M, 10-9 M, 10-8 M, 10-7 M or 10-6 M of Ang II respectively (P < 0.003). The inhibitory effect of Angiotensin II was restored in losartan, an AT1 specific antagonist, pretreated rats, indicating an AT1 mediated effect of Ang II on alveolar fluid clearance. The expression of Na,K-ATPase proteins and cAMP levels in alveolar epithelial cells were down-regulated following the administration of Angiotensin II; suggesting that cAMP may be involved in AngII-induced reduced Na,K-ATPase expression, though the contribution of additional factors could not be excluded. We herein suggest a novel mechanism of clinical relevance by which angiotensin adversely impairs the ability of the lungs to clear edema.


Anti-hyperlipidemic effect of an edible brown algae, Ecklonia stolonifera, and its constituents on poloxamer 407-induced hyperlipidemic and cholesterol-fed rats.
We conducted this study to isolate novel anti-hyperlipidemic agents derived from natural marine products. To accomplish this, we investigated the effects of ethanolic (EtOH) extracts of Ecklonia stolonifera and its phlorotannin constituents, eckol and dieckol, on serum lipid levels in rats with hyperlipidemia that was induced by a high-cholesterol diet or poloxamer 407. Treatment with the EtOH extracts of E. stolonifera and its phlorotannin-rich ethyl acetate (EtOAc) and n-butanol (n-BuOH) fractions induced a significant reduction in triglycerides (TG), total cholesterol (TC), and low-density lipoprotein-cholesterol (LDL-C) levels, as well as a significant increase in the high-density lipoprotein-cholesterol (HDLC) level in hyperlipidemic rats. However, treatment with the water (H(2)O) fraction did not exert any significant effects on the serum levels of hyperlipidemic rats. In addition, eckol and dieckol isolated from the active EtOAc fraction induced a significant reduction in serum TG, TC, and LDL-C levels, as well as in the atherogenic index (A.I.). Furthermore, treatment with dieckol induced a greater decrease in the serum TG, TC, and LDL-C levels of hyperlipidemic rats than eckol or lovastatin, as well as an increase in the serum HDL-C levels. Taken together, these results suggest that phlorotannins such as eckol and dieckol have the potential for use for the prevention of hyperlipidemic atherosclerosis.


Flavonoids in Kidney Health and Disease.
This review summarizes the latest advances in knowledge on the effects of flavonoids on renal function in health and disease. Flavonoids have antihypertensive, antidiabetic, and antiinflammatory effects, among other therapeutic activities. Many of them also exert renoprotective actions that may be of interest in diseases such as glomerulonephritis, diabetic nephropathy, and chemically-induced kidney insufficiency. They affect several renal factors that promote diuresis and natriuresis, which may contribute to their well-known antihypertensive effect. Flavonoids prevent or attenuate the renal injury associated with arterial hypertension, both by decreasing blood pressure and by acting directly on the renal parenchyma. These outcomes derive from their interference with multiple signaling pathways known to produce renal injury and are independent of their blood pressure-lowering effects. Oral administration of flavonoids prevents or ameliorates adverse effects on the kidney of elevated fructose consumption, high fat diet, and types I and 2 diabetes. These compounds attenuate the hyperglycemia-disrupted renal endothelial barrier function, urinary microalbumin excretion, and glomerular hyperfiltration that results from a reduction of podocyte injury, a determinant factor for albuminuria in diabetic nephropathy. Several flavonoids have shown renal protective effects against many nephrotoxic agents that frequently cause acute kidney injury (AKI) or chronic kidney disease (CKD), such as LPS, gentamycin, alcohol, nicotine, lead or cadmium. Flavonoids also improve cisplatin- or methotrexate-induced renal damage, demonstrating important actions in chemotherapy, anticancer and renoprotective effects. A beneficial prophylactic effect of flavonoids has been also observed against AKI induced by surgical procedures such as ischemia/reperfusion (I/R) or cardiopulmonary bypass. In several murine models of CKD, impaired kidney function was significantly improved by the administration of flavonoids from different sources, alone or in combination with stem cells. In humans, cocoa flavanols were found to have vasculoprotective effects in patients on hemodialysis. Moreover, flavonoids develop antitumor activity against renal carcinoma cells with no toxic effects on normal cells, suggesting a potential therapeutic role in patients with renal carcinoma.


UV radiation enhanced oxygen vacancy formation caused by the PLD plasma plume.
Pulsed Laser Deposition is a commonly used non-equilibrium physical deposition technique for the growth of complex oxide thin films. A wide range of parameters is known to influence the properties of the used samples and thin films, especially the oxygen-vacancy concentration. One parameter has up to this point been neglected due to the challenges of separating its influence from the influence of the impinging species during growth: the UV-radiation of the plasma plume. We here present experiments enabled by a specially designed holder to allow a separation of these two influences. The influence of the UV-irradiation during pulsed laser deposition on the formation of oxygen-vacancies is investigated for the perovskite model material SrTiO3. The carrier concentration of UV-irradiated samples is nearly constant with depth and time. By contrast samples not exposed to the radiation of the plume show a depth dependence and a decrease in concentration over time. We reveal an increase in Ti-vacancy-oxygen-vacancy-complexes for UV irradiated samples, consistent with the different carrier concentrations. We find a UV enhanced oxygen-vacancy incorporation rate as responsible mechanism. We provide a complete picture of another influence parameter to be considered during pulsed laser depositions and unravel the mechanism behind persistent-photo-conductivity in SrTiO3.


Transgender men and lactation: what nurses need to know.
Research examining needs of postpartum transgender men in relation to lactation and infant feeding is missing from nursing literature. Accordingly, little is known about how perinatal nurses can best support this unique subset of postpartum patients. Case studies presented here reveal that transgender men would appreciate care from nurses who are knowledgeable about transgender individuals and their healthcare needs, but this type of care is not always available. Nurses need more education about how to best support transgender patients and families in order to achieve optimal lactation and infant nutrition in this population.