Colletotrichum boninense Causing Anthracnose on Coffee Trees in Brazil.
In Brazil, dieback and necrosis of leaves and berries of coffee trees (Coffea arabica and C. canephora) are common symptoms of anthracnose disease caused by Colletotrichum gloeosporioides (Penz.) Sacc. In April 2010, these symptoms were observed in 100% of the plants from different coffee plantations in the Brazilian states of Espírito Santo and Bahia. Ten isolates were obtained from symptomatic leaves and berries from these areas. Of the 10 isolates, one had distinct conidial morphology with hyaline and ellipsoid conidia measuring 10 to 16 × 5.0 to 7.5 μm and melanized irregular or spatulated-shaped appressoria measuring 7.5 to 11.0 × 5.5 to 8.5 μm, formed either solitary or concatenated, which concurred with the conidia description of Colletotrichum boninense. In order to confirm the identity of this isolate, the internal transcribed spacer (ITS) rRNA region and the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene were sequenced (GenBank Accession Nos. JF683320 and JF331654, respectively) and compared to sequences from a database of C. boninense, confirming that the isolate was definitely C. boninense sensu lato, since it was exactly identical to other sequences in a large clade of isolates. To verify the pathogenicity of C. boninense in coffee and to compare the symptoms with those caused by C. gloeosporioides, leaves and berries were inoculated with the isolate of C. boninense and one representative isolate of C. gloeosporioides, both expressing the GFP (green fluorescent protein) gene. The isolates were grown for 7 days on potato dextrose agar and a conidial suspension (106 conidia × ml-1) was used to inoculate the organs, wounded and non-wounded, at different stages of development. In non-wounded organs, the conidial suspension was inoculated on the surface, and in leaves and berries used as control, the suspensions were substituted for sterile water. Leaves and berries were wounded with a sterilized needle and inoculated with 20 and 10 μl of the conidial suspension, respectively. Inoculated materials were incubated at 25°C and 100% relative humidity. The experiment was performed twice and evaluated daily for a week. No symptoms were observed on the control and non-wounded organs, while wounded organs exhibited typical anthracnose symptoms for both species. In berries, C. gloeosporioides consistently caused more severe symptoms at a faster rate than C. boninense. Both fungi caused necrosis in young but not old leaves. Typical acervuli were observed on the lesions and the fungus was successfully recovered from the inoculated tissues, which was confirmed by fluorescence microscopy, fulfilling Koch's Postulates. C. boninense has been identified as a pathogen causing anthracnose in a range of hosts worldwide. However, in Brazil, it has only been reported in pepper (Capsicum annuum) (3), passion fruit (Passiflora) (4), Hippeastrum (1) and in the medicinal plant Maytenus ilicifolia (2). To our knowledge, this is the first report of C. boninense associated with anthracnose of coffee trees in Brazil. Since the symptoms are similar to those caused by C. gloeosporioides, it can be stated that both species are associated with this disease in commercial coffee plantations in Brazil. Therefore, control strategies should consider the occurrence of C. boninense. References: (1) D. F. Farr et al. Mycol. Res. 110:1395, 2006. (2) S. A. Pileggi et al. Can. J. Microbiol. 55:1076, 2009. (3) H. J. Tozze et al. Plant Dis. 93:106, 2009. (4) H. J. Tozze et al. Australas. Plant Dis. Notes 5:70, 2010.


Vasopressin response in collecting ducts of rats resistant to mineralocorticoid hypertension.
In previous studies we found that vasopressin stimulation of both cyclic AMP (cAMP) formation in cortical collecting tubules (CCT) and sodium reabsorption in isolated perfused kidneys was markedly exaggerated in rats with mineralocorticoid hypertension. In the present study, we tested the response (cAMP accumulation) of cortical and outer medullary collecting tubules (OMCT) to vasopressin in two rat models that are resistant to deoxycorticosterone acetate (DOCA)-induced hypertension, the Wistar-Furth strain and NaCl-deficient rats. The blood pressure of normal outbred Wistar rats rose to hypertensive levels (systolic pressure more than 165 mm Hg) during a 5-week treatment with DOCA (10 mg/week) and 1% saline to drink. Significant hypertrophy of the heart and kidneys was also observed. Vasopressin (10(-8) M)-induced cAMP formation was enhanced 3.4-fold in the CCT (OMCT unchanged) of hypertensive rats compared with normotensive controls. Significant hypertrophy (as indexed by tubule diameter) of the CCT but not the OMCT was also observed in DOCA-salt hypertensive rats. Restriction of dietary NaCl (0.13% in chow, tap water to drink) completely prevented DOCA-induced hypertension, organ and CCT hypertrophy, and enhancement of vasopressin-stimulated cAMP formation in the CCT. In Wistar-Furth rats, DOCA-salt treatment did not alter blood pressure or cause significant organ hypertrophy. However, DOCA-salt treatment enhanced vasopressin-stimulated cAMP formation by 4.1-fold in CCT of Wistar-Furth rats, with significant tubular hypertrophy in the CCT but not the OMCT. We conclude that DOCA-induced hypertension and changes in CCT function are dependent on excess dietary NaCl.(ABSTRACT TRUNCATED AT 250 WORDS)


Observational analysis of disparities in obesity in children in the UK: Has Leeds bucked the trend?
The prevalence of obesity in childhood is of high concern, especially in deprived populations. We explored trends in obesity following the introduction of a citywide strategy focused on preschool children. Analysis of obesity prevalence using the National Child Measurement Programme 2009 to 2017 for primary-school children in Leeds using 5-year aggregated data for Leeds, comparable cities, and England as a whole. Prevalence of obesity in Leeds for school entry children fell significantly (9.4% to 8.8%), whilst comparable cities (CC) and England as a whole showed no change (comparison of trends: P < 0.001 and P < 0.001). The reduction in Leeds was primarily in the most deprived (11.5% to 10.5%; trend comparison CC: P < 0.001, Eng: P < 0.001), but also amongst the affluent (6.8% to 6.0%; trend comparison CC: P = 0.087, Eng: P = 0.012). Prevalence in older children in Leeds was unchanged whilst it increased for comparable cities and England (trend comparison CC: P < 0.001, Eng: P < 0.001). In the deprived, obesity increased: Leeds by 1.4%; CC 1.3%, England 1% (trend comparison Eng: P = 0.004). In the affluent, obesity prevalence reduced more in Leeds than elsewhere: 2% in Leeds, 0.8% in CC, and 0.7% in England (trend comparison CC: P < 0.001, Eng: P ≤ 0.001). There has been a notable decrease in the prevalence of obesity especially amongst the most disadvantaged children at entry to primary school in Leeds. How this was achieved merits in-depth consideration.


Accumulation of inositol phosphates in sympathetic ganglia. Effects of depolarization and of amine and peptide neurotransmitters.
Depolarization of isolated [3H]inositol-labelled rat superior cervical sympathetic ganglia in a high-K+ medium stimulates an accumulation of labelled inositol phosphates. This accumulation occurs only when ganglia are incubated in a Ca2+-containing medium, suggesting that it represents a receptor-stimulated hydrolysis of inositol lipid(s) activated by an endogenously released neurotransmitter. A minor fraction of this accumulation appears to be activated by intraganglionically released acetylcholine, since it is slightly reduced by atropine. The accumulation of inositol phosphates is unaffected by blockade of appropriate catecholamine, histamine and 5-hydroxytryptamine receptors and also by aspirin and indomethacin. This response to depolarization is potentiated by incubation with proteinase inhibitors, suggesting that it might be caused by an endogenously released peptide neutrotransmitter. However, it is not prevented by a V1-vasopressin receptor antagonist, and none of the peptides tested so far fully reproduces the response: these include a stable substance P analogue, physalaemin, neurokinin alpha, bradykinin, angiotensin, pancreozymin, bombesin and luteinizing-hormone-releasing hormone. Stimulated inositol lipid breakdown in depolarized sympathetic ganglia seems likely to be activated by an as-yet-unidentified peptide neurotransmitter: this might serve as an intraganglionic mediator of postsynaptic excitation by employing the same signalling mechanism as muscarinic cholinergic and V1-vasopressin receptors.


Extended robust support vector machine based on financial risk minimization.
Financial risk measures have been used recently in machine learning. For example, ν-support vector machine ν-SVM) minimizes the conditional value at risk (CVaR) of margin distribution. The measure is popular in finance because of the subadditivity property, but it is very sensitive to a few outliers in the tail of the distribution. We propose a new classification method, extended robust SVM (ER-SVM), which minimizes an intermediate risk measure between the CVaR and value at risk (VaR) by expecting that the resulting model becomes less sensitive than ν-SVM to outliers. We can regard ER-SVM as an extension of robust SVM, which uses a truncated hinge loss. Numerical experiments imply the ER-SVM's possibility of achieving a better prediction performance with proper parameter setting.


Maps and metrics of insecticide-treated net access, use, and nets-per-capita in Africa from 2000-2020.
Insecticide-treated nets (ITNs) are one of the most widespread and impactful malaria interventions in Africa, yet a spatially-resolved time series of ITN coverage has never been published. Using data from multiple sources, we generate high-resolution maps of ITN access, use, and nets-per-capita annually from 2000 to 2020 across the 40 highest-burden African countries. Our findings support several existing hypotheses: that use is high among those with access, that nets are discarded more quickly than official policy presumes, and that effectively distributing nets grows more difficult as coverage increases. The primary driving factors behind these findings are most likely strong cultural and social messaging around the importance of net use, low physical net durability, and a mixture of inherent commodity distribution challenges and less-than-optimal net allocation policies, respectively. These results can inform both policy decisions and downstream malaria analyses.


Role of complement and the Fc portion of immunoglobulin G in immunity to Venezuelan equine encephalomyelitis virus infection with glycoprotein-specific monoclonal antibodies.
We have previously characterized with monoclonal antibodies (MAbs) seven unique epitopes on the two envelope glycoproteins of Venezuelan equine encephalomyelitis (VEE) virus vaccine strain TC-83. The epitopes important in protection from VEE virus infection were determined in passive antibody transfer studies, with virulent VEE (Trinidad donkey) virus as the challenge virus. Selected high-avidity MAbs to the three major protective epitopes (E2c, E1b, and E1d) were assayed for in vitro complement activity. All three fixed murine complement to high titer. Limited pepsin digestion of the anti-E2c in the presence of cysteine resulted in a rapid decrease and complete loss of complement-fixing ability by 2 h, but the majority of mice, except at the lowest dilution of MAb, were protected until the Fc termini were cleaved at 3 h. Anti-E2c F(ab')2 would neutralize VEE (Trinidad donkey) virus more efficiently than either Fab' or Fab; none of the fragments would fix complement or was effective in passive protection. C5-deficient mice and mice depleted of C3 with cobra venom factor were still protected from VEE (Trinidad donkey) virus challenge after passive transfer of either anti-E2c or anti-E1b MAb. The results show that the anti-E2c MAb mediates neutralization through bivalent binding at a critical site on the virion and that Fc effector functions, other than complement, are necessary for protection. Although the ability of the anti-E2c MAb to fix complement was associated with its ability to protect in vivo, no direct cause-and-effect relationship was found. Since the epitope defined by the anti-E1d antibody is found on the cell membrane, but is not expressed on the infectious virion, protection in mice was most likely mediated at the cellular level, possibly by inhibition of the final stages of virion maturation.


[Refractive surgery].
The surgical correction of refractive errors has gained widespread acceptance in the past 20 years, mainly through the introduction of the excimer laser. The excimer is used to ablate the cornea, renoving stromal tissue in the center (to correct myopia through flattening of the surface), or in the midperiphery (to steepen the cornea to correct hyperopia). Although excimer procedures dominate the field of refractive surgery, other approaches are also available. In the cornea itself, ring segments can be implanted, heat-induced coagulation effects can produce steepening, and cross-linking of the collagen fibers can stiffen a weakened structure. While all corneal procedures are extraocular, refractive surgery can also be performed within the eye. Special intraocular lenses can be implanted in the anterior chamber angle, fixated onto the iris, or placed in the posterior chamber in front of the crystalline lens. These so-called phakic intraocular lenses are available in different optical magnitudes and act to correct the patient's refractive error. Prebyopic individuals can have the crystalline lens removed in a procedure identical to cataract surgery. In such cases the lens is extracted even though no cataract is present, and an intraocular lens is implanted in the now empty capsular bag of the crystalline lens. The implanted lens serves to correct the preoperative refractive error.


Nuclear autoantigenic sperm protein (NASP), a linker histone chaperone that is required for cell proliferation.
A multichaperone nucleosome-remodeling complex that contains the H1 linker histone chaperone nuclear autoantigenic sperm protein (NASP) has recently been described. Linker histones (H1) are required for the proper completion of normal development, and NASP transports H1 histones into nuclei and exchanges H1 histones with DNA. Consequently, we investigated whether NASP is required for normal cell cycle progression and development. We now report that without sufficient NASP, HeLa cells and U2OS cells are unable to replicate their DNA and progress through the cell cycle and that the NASP(-/-) null mutation causes embryonic lethality. Although the null mutation NASP(-/-) caused embryonic lethality, null embryos survive until the blastocyst stage, which may be explained by the presence of stored NASP protein in the cytoplasm of oocytes. We conclude from this study that NASP and therefore the linker histones are key players in the assembly of chromatin after DNA replication.


Fasting proinsulin concentrations predict the development of type 2 diabetes.
The development of specific assays allows the different molecules in the proinsulin processing pathway to be measured separately. 32,33 Split proinsulin is the predominant form of proinsulin and accounts for the disproportionate hyperproinsulinemia seen in individuals with prevalent type 2 diabetes. This study was established to examine whether the concentration of this molecule predicts diabetes. A population-based longitudinal cohort study was conducted in Ely, Cambridgeshire. At baseline, 1,122 individuals completed a 75-g oral glucose tolerance test (OGTT). At the 4.5-year follow-up study, repeat OGTTs were performed on 937 of the cohort of 1,071 individuals who had been nondiabetic at baseline. A total of 26 people progressed to diabetes as determined by the OGTTs. The risk of progression was strongly related to the fasting glucose concentration (relative risk [RR] comparing top with bottom quartile 17.6 [95% CI 2.4-130.4]) and fasting 32,33 split proinsulin (RR 16.4 [2.2-121.9]), but less strongly to the fasting insulin (RR 4.41 [1.5-12.9]) or intact proinsulin (RR 5.2 [1.5-17.3]). In multivariate analyses, these associations were independent of age, sex, BMI, and baseline glucose tolerance category. Subjects in the top quartile for fasting glucose and total proinsulin with a family history of diabetes were a high-risk subgroup (incidence 65.8 per 1,000 person-years of follow-up [pyfu]); 30% of them progressed to diabetes at follow-up. Fasting 32,33 split proinsulin independently predicts the development of diabetes. This prediction was better than that observed for either the insulin or intact proinsulin concentrations. The combination of family history, fasting glucose, and total proinsulin identified a subgroup of individuals at high risk of progression who might benefit from targeted interventions.


Source of raised serum estrogens in male rats with portal bypass.
We sought to establish the mechanism for the raised serum estrogen levels that occur in male rats with portal hypertension and resultant portal bypass. Using the portal vein ligated (PVL) rat model, we evaluated plasma steroid hormone concentrations, metabolic clearance rate (MCR) of estradiol, and hepatic metabolism of androstenedione to estrogens and other products. In contrast to serum testosterone levels that were reduced, serum androstenedione levels were normal in the PVL rat. Estradiol MCR was measured by a constant intravenous infusion technique and was found to be similar in PVL and control animals. Androstenedione MCR was determined during constant intravenous infusion of [3H]androstenedione, and the resultant radiolabeled steroids present in plasma were separated by thin layer chromatography. The MCR of androstenedione was not diminished in PVL rats compared with controls. However, there was a sevenfold increase in the plasma estradiol derived from [3H]androstenedione in rats with portal bypass. Examination of radiolabel excreted in bile during infusion of [3H]androstenedione showed that 25-46% of this steroid was converted to estradiol in PVL rats compared with less than 3% in control male rats (P less than 0.001). Moreover, there was a selective reduction in the excretion of 16 alpha-hydroxyandrostenedione, a finding which suggested that the metabolism of androstenedione via this pathway was decreased. Androstenedione 16 alpha-hydroxylation is known to be catalyzed by a male-specific cytochrome P-450 isoform, P-450UT-A. We conclude that raised plasma estradiol levels after portal bypass in male rats are due to increased production rates, resulting in turn from enhanced aromatization of androstenedione to estradiol. On the basis of the observed specific changes in androstenedione hydroxylation pathways, it is proposed that alterations in levels of sex-specific forms of cytochrome P-450 occur in male rats with portal bypass and could account for the enhanced formation of estradiol.


Contribution of IVIM to Conventional Dynamic Contrast-Enhanced and Diffusion-Weighted MRI in Differentiating Benign from Malignant Breast Masses.
The aim of this study was to determine whether the indicators obtained from intravoxel incoherent motion (IVIM) imaging can improve the characterization of benign and malignant breast masses compared with conventional dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) and diffusion-weighted magnetic resonance imaging (DW-MRI). This study included 23 benign and 31 malignant breast masses of 48 patients. Main indicators were initial enhancement ratio (IER), time-signal intensity curve (TIC), apparent diffusion coefficient (ADC), tissue diffusivity (D), pseudodiffusivity (D*), and perfusion fraction (f). The discriminative abilities of the different models were compared by means of receiver operating characteristic (ROC) curve and area under the ROC curve (AUC) analysis. D had the highest AUC (0.980), sensitivity (93.55%), specificity (100%), and diagnostic accuracy (96.36%). Both D and TIC could provide the independent predicted features for malignant breast masses. The combination of D and TIC had an AUC of up to 0.990. D of IVIM can effectively complement existing conventional DCE-MRI and DW-MRI in differentiating malignant from benign breast masses. IVIM combined with DCE-MRI is a robust means of evaluating breast masses.


Sizing the mitral annulus in healthy subjects and patients with mitral regurgitation: 2D versus 3D measurements from cardiac CT.
The purpose of our study was (1) to assess retrospectively, in healthy subjects and in patients with moderate and severe functional mitral regurgitation (FMR), the normal mitral annular dimensions, (2) to determine differences in mitral annular geometry between healthy subjects and patients with FMR, and (3) to evaluate potential errors in 2-dimensional (2D) measurements given the 3D nature of the mitral annulus. 15 patients with no cardiac abnormalities (referred to as normals), 13 with moderate and 15 with severe FMR as determined by echocardiography underwent contrast-enhanced cardiac 64-slice Computed tomography (CT) with prospective electrocardiography-gating for excluding coronary artery disease. With an advanced visualization, segmentation, and image analysis software, the area, intercommissural distance (CC), septolateral distance (SLD), and the anterior and posterior circumference of the MA were measured in diastole. We found significant (P < .001) differences between normals and patients with severe FMR for area, SLD and posterior circumference in 3D (P < .001) and 2D (P < .001). Similarly, the SLD and the posterior circumference in both 3D (P = .002) and 2D (P = .001) were significantly smaller in patients with moderate FMR as compared to those with severe FMR. In contrast, there were no significant differences between groups regarding the CC and the anterior circumference both in 3D and 2D (all, P > .05). Measurements in 3D differed significantly from those with 2D for all circumference measurements and groups (P < .01), with a systematic underestimation of the posterior circumference of 2.1 ± 1.5 mm in normals, 1.8 ± 1.3 mm in patients with moderate FMR, and 1.9 ± 1.9 mm in patients with severe FMR for 2D. Our study provides in vivo human CT data on MA dimensions in normals and patients with FMR, indicating differences in patients for the area, posterior circumference and SLD but not for the anterior circumference and CC. Systematic differences exist between 2D and 3D measurements for all circumferential measurements.


Generalization of an Early Intervention for Inhibited Preschoolers to the Classroom Setting.
Considerable evidence has accumulated supporting transactional influences between early childhood behavioral inhibition (BI), parent-child and child-peer relationships, and the development of anxiety disorders in adolescence and adulthood. Drawing from this literature, the Turtle Program was designed to treat children high in BI by intervening at the level of both parents and peers. In this pilot study, we sought to determine whether benefits of participating in the Turtle Program extended to children's classrooms in the form of increased positive social interactions with peers. Forty inhibited children (42-60 months) and their parent(s) were randomized to either the Turtle Program (n = 18) or a waitlist control group (WLC; n = 22). The Turtle Program involved 8 weeks of concurrent parent and child treatment. Trained research assistants, blind to treatment condition, coded participants' social interactions with peers during free play at each child's preschool at the beginning and end of treatment. Teachers unaware of group assignment also provided reports of social behaviors at these time points. Reliable change index scores revealed that both Turtle Program and WLC participants experienced relatively high rates of reliable increases in observed peer play interactions from pre- to post-treatment (73.3% and 42.1% respectively). Additionally, Turtle Program participants experienced high rates of reliable increase in observed initiations to peers (73.3%) as well as a moderate degree of reliable decrease in teacher-reported displays of fear/anxiety (33.3%). These data provide preliminary, but promising, evidence that increases in children's social behaviors as a result of participation in the Turtle Program generalize to their preschool classrooms.


Patient perspectives on physician conflict of interest in industry-sponsored clinical trials for multiple sclerosis therapeutics.
Pharmaceutical industry financial support of physicians, physician practices, and academic departments involved in multicenter industry-sponsored clinical trials of novel therapeutic agents is a relatively new and infrequently acknowledged source of potential physician conflict of interest. Detailed disclosure of these relationships to study participants is not uniformly a part of informed consent and documentation practices. To understand attitudes of patients with multiple sclerosis concerning disclosure of potential physician-industry conflicts of interest created by clinical trials and how such disclosures may influence study participation An anonymous online instrument was developed. 597 people with multiple sclerosis participated in the study. The study found that detailed disclosure of conflicts of interest is important to potential participants in industry-sponsored clinical trials for multiple sclerosis therapies and that the presence of these conflicts of interest may influence patients' decisions to participate in these studies. Findings from this study support a call for uniform guidelines regarding disclosure of physician-industry relationships to prospective research participants for industry-sponsored clinical trials.


Ascl1/Mash1 is a novel target of Gli2 during Gli2-induced neurogenesis in P19 EC cells.
The Sonic Hedgehog (Shh) signaling pathway is important for neurogenesis in vivo. Gli transcription factors, effector proteins of the Shh signaling pathway, have neurogenic properties in vivo, which are still poorly understood. To study the molecular basis of neurogenic properties of Gli2, we used a well-established embryonic stem cell model, the P19 embryonal carcinoma (EC) cell line, which can be induced to differentiate into neurons in the presence of retinoic acid (RA). We found that, in the absence of RA, overexpression of Gli2 induced P19 EC cells to differentiate into neurons, but not astrocytes during the first ten days of differentiation. To our knowledge, this is the first indication that the expression of Gli factors can convert EC cells into neurons. Furthermore, Gli2 upregulated expression of the neurogenic basic helix-loop-helix (bHLH) factors, such as NeuroD, Neurog1 and Ascl1/Mash1 in P19 EC cells. Using chromatin immunoprecipitation assays, we showed that Gli2 bound to multiple regulatory regions in the Ascl1 gene, including promoter and enhancer regions during Gli2-induced neurogenesis. In addition, Gli2 activated the Ascl1/Mash1 promoter in vitro. Using the expression of a dominant-negative form of Gli2, fused to the Engrailed repression domain, we observed a reduction in gliogenesis and a significant downregulation of the bHLH factors Ascl1/Mash1, Neurog1 and NeuroD, leading to delayed neurogenesis in P19 EC cells, further supporting the hypothesis that Ascl1/Mash1 is a direct target of Gli2. In summary, Gli2 is sufficient to induce neurogenesis in P19 stem cells at least in part by directly upregulating Ascl1/Mash1. Our results provide mechanistic insight into the neurogenic properties of Gli2 in vitro, and offer novel plausible explanations for its in vivo neurogenic properties.


Proteome analyses of strains ATCC 51142 and PCC 7822 of the diazotrophic cyanobacterium Cyanothece sp. under culture conditions resulting in enhanced H₂ production.
Cultures of the cyanobacterial genus Cyanothece have been shown to produce high levels of biohydrogen. These strains are diazotrophic and undergo pronounced diurnal cycles when grown under N(2)-fixing conditions in light-dark cycles. We seek to better understand the way in which proteins respond to these diurnal changes, and we performed quantitative proteome analysis of Cyanothece sp. strains ATCC 51142 and PCC 7822 grown under 8 different nutritional conditions. Nitrogenase expression was limited to N(2)-fixing conditions, and in the absence of glycerol, nitrogenase gene expression was linked to the dark period. However, glycerol induced expression of nitrogenase during part of the light period, together with cytochrome c oxidase (Cox), glycogen phosphorylase (Glp), and glycolytic and pentose phosphate pathway (PPP) enzymes. This indicated that nitrogenase expression in the light was facilitated via higher levels of respiration and glycogen breakdown. Key enzymes of the Calvin cycle were inhibited in Cyanothece ATCC 51142 in the presence of glycerol under H(2)-producing conditions, suggesting a competition between these sources of carbon. However, in Cyanothece PCC 7822, the Calvin cycle still played a role in cofactor recycling during H(2) production. Our data comprise the first comprehensive profiling of proteome changes in Cyanothece PCC 7822 and allow an in-depth comparative analysis of major physiological and biochemical processes that influence H(2) production in both strains. Our results revealed many previously uncharacterized proteins that may play a role in nitrogenase activity and in other metabolic pathways and may provide suitable targets for genetic manipulation that would lead to improvement of large-scale H(2) production.


Electrophysiological and behavioral responses of Dendroctonus frontalis (Coleoptera: Curculionidae) to volatiles isolated from conspecifics.
Olfactory sensitivity of the southern pine beetle, Dendroctonus frontalis Zimmermann, to compounds isolated from the mid/hindguts of newly emerged conspecific adults was assayed with coupled gas chromatography-electroantennographic detection. All previously reported pheromones for D. frontalis plus eight additional compounds (fenchyl alcohol, myrtenal, cis-verbenol, trans-pinocarveol, acetophenone, trans-myrtanol, cis-myrtanol, and 2-phenylethanol) consistently elicited antennal responses from at least one sex. The eight additional compounds were assayed individually at three release rates (0.4-0.8, 3-9, and 25-100 mg/d) for the ability to alter D. frontalis responses to traps baited with D. frontalis attractant (4 mg/d frontalin and 17 mg/d alpha-pinene). At the high release rate, cis-verbenol enhanced attraction of D. frontalis females, whereas the other seven compounds significantly reduced attraction of one or both sexes. Acetophenone significantly reduced attraction of male D. frontalis at the low release rate, and five compounds (fenchyl alcohol, trans-pinocarveol, acetophenone, cis-myrtanol, and 2-phenylethanol) reduced attraction of one or both sexes at the intermediate rate. Only acetophenone significantly altered the sex ratio of beetles trapped, decreasing the proportion of males. Attraction of predatory checkered beetles (Cleridae) was enhanced by cis-verbenol released at the high rate but was not altered by any compound inhibitory to D. frontalis. Analyses of volatiles from individual D. frontalis indicated that the majority of the eight compounds were produced in greater quantities by newly emerged beetles than ones attacking pine bolts. Five of the compounds were associated predominantly with one sex. Possible ecological roles of these compounds in the biology of D. frontalis are discussed.


Maternal mental health symptoms are positively related to emotional and restrained eating attitudes in a statewide sample of mothers participating in a supplemental nutrition program for women, infants and young children.
Postpartum, low-income mothers are at risk for mental health symptoms and obesity, and disordered eating attitudes may be associated with both mental health and obesity in this vulnerable population. The study objective is to determine whether higher levels of mental health symptoms are associated with increased odds of emotional and restrained eating attitudes in this sample of Special Supplemental Nutrition Program for Women, Infants and Children (WIC) participants. Data on 711 mothers of infants <13 months from a statewide sample of Maryland WIC participants were collected via telephone survey. Maternal mental health symptoms were measured on continuous scales for depression (PRIME-MD), stress (Perceived Stress Scale) and anxiety (Spielberger State-Trait Anxiety Inventory). Emotional and restrained eating attitudes were measured with questions adapted from the Dutch Eating Behavior Questionnaire. Multivariate logistic regression analysis was used. Obesity [body mass index (BMI) ≥ 30] was explored as a moderating variable. Mothers reporting higher levels of depression symptoms [odds ratio (OR) = 3.93, 95%CI: 2.71-5.69], anxiety symptoms (OR = 1.96, 95%CI: 1.47-2.65), stress symptoms (OR = 2.09, 95%CI: 1.67-2.61) and high overall mental health symptomatology (OR = 3.51, 95%CI: 2.43-5.3) had increased odds of emotional eating attitudes. There were significant associations between symptoms of depression (OR = 1.59, 95% CI: 1.12-2.25) and increased odds of restrained eating attitudes. Obesity did not moderate the association. Mothers with mental health symptoms are at risk for disordered eating attitudes, which may increase risk of poor diet. These findings underscore the need for greater focus on addressing maternal mental health status and eating attitudes in the postpartum period.


Cannabinoids induce apoptosis of pancreatic tumor cells via endoplasmic reticulum stress-related genes.
Pancreatic adenocarcinomas are among the most malignant forms of cancer and, therefore, it is of especial interest to set new strategies aimed at improving the prognostic of this deadly disease. The present study was undertaken to investigate the action of cannabinoids, a new family of potential antitumoral agents, in pancreatic cancer. We show that cannabinoid receptors are expressed in human pancreatic tumor cell lines and tumor biopsies at much higher levels than in normal pancreatic tissue. Studies conducted with MiaPaCa2 and Panc1 cell lines showed that cannabinoid administration (a) induced apoptosis, (b) increased ceramide levels, and (c) up-regulated mRNA levels of the stress protein p8. These effects were prevented by blockade of the CB(2) cannabinoid receptor or by pharmacologic inhibition of ceramide synthesis de novo. Knockdown experiments using selective small interfering RNAs showed the involvement of p8 via its downstream endoplasmic reticulum stress-related targets activating transcription factor 4 (ATF-4) and TRB3 in Delta(9)-tetrahydrocannabinol-induced apoptosis. Cannabinoids also reduced the growth of tumor cells in two animal models of pancreatic cancer. In addition, cannabinoid treatment inhibited the spreading of pancreatic tumor cells. Moreover, cannabinoid administration selectively increased apoptosis and TRB3 expression in pancreatic tumor cells but not in normal tissue. In conclusion, results presented here show that cannabinoids lead to apoptosis of pancreatic tumor cells via a CB(2) receptor and de novo synthesized ceramide-dependent up-regulation of p8 and the endoplasmic reticulum stress-related genes ATF-4 and TRB3. These findings may contribute to set the basis for a new therapeutic approach for the treatment of pancreatic cancer.


Cointegration analysis of vulnerability index and standardised precipitation index in Mafeteng district, Lesotho.
Given the high poverty levels in Africa, with most countries' economy and populations' livelihood dependent on rain-fed agriculture, land degradation among other environmental hazards has proven to be a major threat to economic growth and food insecurity, respectively. Drought, which is on the increase at the global level and said to create over 78% of other hazards, has aggravated land degradation. Dry conditions lessen soil particles cohesion force, thereby increasing susceptibility of such soils to be lost by wind and water. The current study aimed at estimating land degradation from drought hazard index, standardised precipitation index (SPI) over the drought declared district of Mafeteng Lesotho. Data were provided by Lesotho Meteorological Services for a period of 30 years (1984-2014). All missing values that existed in the collected precipitation data were filled with average values of the months with data. The computation of SPI was performed by using DrinC software in SPI-3 and SPI-Annual time step. The results revealed a constant condition of land degradation vulnerability over a 30-year period, implying a continuous loss of soil fertility, agricultural gross domestic product (GDP), water and bio-energy, malnutrition and increased poverty levels.


Chronology of martian breccia NWA 7034 and the formation of the martian crustal dichotomy.
Martian meteorite Northwest Africa (NWA) 7034 and its paired stones are the only brecciated regolith samples from Mars with compositions that are representative of the average martian crust. These samples therefore provide a unique opportunity to constrain the processes of metamorphism and alteration in the martian crust, which we have investigated via U-Pu/Xe, 40Ar/39Ar, and U-Th-Sm/He chronometry. U-Pu/Xe ages are comparable to previously reported Sm-Nd and U-Pb ages obtained from NWA 7034 and confirm an ancient (>4.3 billion years) age for the source lithology. After almost 3000 million years (Ma) of quiescence, the source terrain experienced several hundred million years of thermal metamorphism recorded by the K-Ar system that appears to have varied both spatially and temporally. Such protracted metamorphism is consistent with plume-related magmatism and suggests that the source terrain covered an areal extent comparable to plume-fed edifices (hundreds of square kilometers). The retention of such expansive, ancient volcanic terrains in the southern highlands over billions of years suggests that formation of the martian crustal dichotomy, a topographic and geophysical divide between the heavily cratered southern highlands and smoother plains of the northern lowlands, likely predates emplacement of the NWA 7034 source terrain-that is, it formed within the first ~100 Ma of planetary formation.


Forward ray tracing for image projection prediction and surface reconstruction in the evaluation of corneal topography systems.
A forward ray tracing (FRT) model is presented to determine the exact image projection in a general corneal topography system. Consequently, the skew ray error in Placido-based topography is demonstrated. A quantitative analysis comparing FRT-based algorithms and Placido-based algorithms in reconstructing the front surface of the cornea shows that arc step algorithms are more sensitive to noise (imprecise). Furthermore, they are less accurate in determining corneal aberrations particularly the quadrafoil aberration. On the other hand, FRT-based algorithms are more accurate and more precise showing that point to point corneal topography is superior compared to its Placido-based counterpart.


Conformational remodeling of femtomolar inhibitor-acetylcholinesterase complexes in the crystalline state.
The active center of acetylcholinesterase (AChE), a target site for competitive inhibitors, resides centrosymmetric to the subunit at the base of a deep, narrow gorge lined by aromatic residues. At the gorge entry, a peripheral site encompasses overlapping binding loci for noncompetitive inhibitors, which alter substrate access to the gorge. The click-chemistry inhibitor TZ2PA6 links the active center ligand, tacrine, to the peripheral site ligand, propidium, through a biorthogonal reaction of an acetylene and an azide that forms either a syn1 or an anti1 triazole. Compared with wild-type mouse AChE, a Tyr337Ala mutant displays full catalytic activity, albeit with 2-3 orders of magnitude higher affinities for the TZ2PA6 syn1 and anti1 regioisomers, reflected in low femtomolar K(d) values, diffusion-limited association, and dissociation half-times greater than 1 month and 1 week, respectively. Three structures of each of the co-crystallized syn1 and anti1 complexes of the Tyr337Ala mutant were solved at three distinct times of crystal maturation, consistent with or exceeding the half-lives of the complexes in solution, while crystalline complexes obtained from soaked Tyr337Ala crystals led to picturing "freshly formed" complexes. The structures, at 2.55-2.75 Å resolution, reveal a range of unprecedented conformations of the bound regioisomers, not observed in the wild-type AChE complexes, associated with concerted positional rearrangements of side chains in the enzyme gorge. Moreover, time-dependent conformational remodeling of the crystalline complexes appears to correlate with the dissociation half-times of the solution complexes. Hence, for the tight-binding TZ2PA6 inhibitors, the initial complexes kinetically driven in solution slowly form more stable complexes governed by thermodynamic equilibrium and observable in mature crystals.


Boron-containing delocalised lipophilic cations for the selective targeting of cancer cells.
To limit the incidence of relapse, cancer treatments must not promote the emergence of drug resistance in tumour and cancer stem cells. Under the proviso that a therapeutic amount of boron is selectively delivered to cancer cells, Boron Neutron Capture Therapy (BNCT) may represent one approach that meets this requirement. To this end, we report the synthesis and pharmacology of several chemical entities, based on boron-rich carborane moieties that are functionalised with Delocalized Lipophilic Cations (DLCs), which selectively target the mitochondria of tumour cells. The treatment of tumour and cancer stem cells (CSCs) with such DLC-functionalized carboranes (DLC-carboranes) induces cell growth arrest that is both highly cancer-cell-selective and permanent. Experiments involving cultures of normal and cancer cells show that only normal cells exhibit recapitulation of their proliferation potential upon removal of the DLC-carborane treatment. At the molecular level, the pharmacological effect of DLC-carboranes is exerted through activation of the p53/p21 axis.


HIV-1 clade B pol evolution following primary infection.
Characterize intra-individual HIV-1 subtype B pol evolution in antiretroviral naive individuals. Longitudinal cohort study of individuals enrolled during primary infection. Eligible individuals were antiretroviral naïve participants enrolled in the cohort from December 1997-December 2005 and having at least two blood samples available with the first one collected within a year of their estimated date of infection. Population-based pol sequences were generated from collected blood samples and analyzed for genetic divergence over time in respect to dual infection status, HLA, CD4 count and viral load. 93 participants were observed for a median of 1.8 years (Mean = 2.2 years, SD =1.9 years). All participants classified as mono-infected had less than 0.7% divergence between any two of their pol sequences using the Tamura-Nei model (TN93), while individuals with dual infection had up to 7.0% divergence. The global substitution rates (substitutions/nucleotide/year) for mono and dually infected individuals were significantly different (p<0.001); however, substitution rates were not associated with HLA haplotype, CD4 or viral load. Even after a maximum of almost 9 years of follow-up, all mono-infected participants had less than 1% divergence between baseline and longitudinal sequences, while participants with dual infection had 10 times greater divergence. These data support the use of HIV-1 pol sequence data to evaluate transmission events, networks and HIV-1 dual infection.


End of life care in Wales: evaluation of a care pathway-based implementation strategy.
The All Wales Care Pathway for the End of Life Pathway aims to ensure evidence-based end-of-life care is available across Wales. Implementation of end-of-life care processes was evaluated in a national audit of deaths between July 2007 and June 2009. Up to 60 records of deceased patients were reviewed by two researchers from hospital (24), community localities (20), hospice and specialist inpatient (9) settings. Data extraction using a standard template was carried out at all sites to indicate whether end-of-life care processes had been implemented. A total of 1184 records were retrieved. 202 records were excluded due to sudden death (eg, cardiac arrest) or incomplete data. Sampling included 580 decedents (59%) who had received end-of-life care through the pathway. Pathway use was associated with improved implementation of all evidence-based clinical standards other than for daily review, where implementation was consistently high with (84.5%) or without the pathway (81%). Differences in achievement were most evident for the implementation of bereavement and spiritual support where the pathway was used. Implementation within hospice and specialist inpatient care settings was consistently high. Integrated care pathway use is associated with the implementation of best practice in end-of-life care. However, variation in implementation across sites and the influence of setting type highlights the mediating effect of organisational context which, together with different methods of feedback, may provide a useful agenda for implementation research within end-of-life care.


Profiling the vendors of COVID-19 related product on the Darknet: An observational study.
In a time of unprecedented global change, the COVID-19 pandemic has led to a surge in demand of COVID-19 vaccines and related certifications. Mainly due to supply shortages, counterfeit vaccines, fake documentation, and alleged cures to illegal portfolios, have been offered on darkweb marketplaces (DWMs) with important public health consequences. We aimed to profile key DWMs and vendors by presenting some in-depth case studies. A non-systematic search for COVID-19 products was performed across 118 DWMs. Levels of activity, credibility, content, COVID-19 product listings, privacy protocols were among the features retrieved. Open web fora and other open web sources were also considered for further analysis of both functional and non functional DWMs. Collected data refers to the period between January 2020 and October 2021. A total of 42 relevant listings sold by 24 vendors across eight DWMs were identified. Four of these markets were active and well-established at the time of the study with good levels of credibility. COVID-19 products were listed alongside other marketplace content. Vendors had a trusted profile, communicated in English language and accepted payments in cryptocurrencies (Monero or Bitcoin). Their geographical location included the USA, Asia and Europe. While COVID-19 related goods were mostly available for regional supply, other listings were also shipped worldwide. Findings emerging from this study rise important questions about the health safety of certain DWMs activities and encourage the development of targeted interventions to overcome such new and rapidly expanding public health threats. CovSaf, National Research centre on Privacy, Harm Reduction and Adversarial Influence Online (REPHRAIN), Commonwealth Fund.


Assessment With Children Who Need Augmentative and Alternative Communication (AAC): Clinical Decisions of AAC Specialists.
The purpose of this study was to explore how speech-language pathologists (SLPs) who are augmentative and alternative communication (AAC) specialists approach the assessment process for 2 case studies, 1 child with cerebral palsy and 1 with autism spectrum disorder. The aim of the study was to answer the following questions: (a) How do clinicians with expertise approach the AAC assessment process for children with developmental disabilities? (b) Can any initial hypothesis be drawn about how SLPs approach the assessment of children with motor versus social interactive deficits? This study used a phenomenological qualitative design. The researchers conducted 2 in-depth, semistructured interviews with 8 SLPs who specialized in AAC and self-identified as primarily working with children. Four major themes emerged from the data: area of assessment, method of assessment, evaluation preparation, and parent education. Each major theme contained multiple subthemes and categories within those subthemes. Participants discussed similar areas of assessment for both cases, indicating that some aspects of AAC assessment are universal. However, the specific aspects of what they were assessing and how they went about assessing them differed between the 2 cases. The results of the current study provide an outline of an assessment protocol for children with complex communication needs.


Histone acetylation in neuronal (dys)function.
Cognitive functions require the expression of an appropriate pattern of genes in response to environmental stimuli. Over the last years, many studies have accumulated knowledge towards the understanding of molecular mechanisms that regulate neuronal gene expression. Epigenetic modifications have been shown to play an important role in numerous neuronal functions, from synaptic plasticity to learning and memory. In particular, histone acetylation is a central player in these processes. In this review, we present the molecular mechanisms of histone acetylation and summarize the data underlying the relevance of histone acetylation in cognitive functions in normal and pathological conditions. In the last part, we discuss the different mechanisms underlying the dysregulation of histone acetylation associated with neurological disorders, with a particular focus on environmental causes (stress, drugs, or infectious agents) that are linked to impaired histone acetylation.


Differential responses of herbivores and herbivory to management in temperate European beech.
Forest management not only affects biodiversity but also might alter ecosystem processes mediated by the organisms, i.e. herbivory the removal of plant biomass by plant-eating insects and other arthropod groups. Aiming at revealing general relationships between forest management and herbivory we investigated aboveground arthropod herbivory in 105 plots dominated by European beech in three different regions in Germany in the sun-exposed canopy of mature beech trees and on beech saplings in the understorey. We separately assessed damage by different guilds of herbivores, i.e. chewing, sucking and scraping herbivores, gall-forming insects and mites, and leaf-mining insects. We asked whether herbivory differs among different forest management regimes (unmanaged, uneven-aged managed, even-aged managed) and among age-classes within even-aged forests. We further tested for consistency of relationships between regions, strata and herbivore guilds. On average, almost 80% of beech leaves showed herbivory damage, and about 6% of leaf area was consumed. Chewing damage was most common, whereas leaf sucking and scraping damage were very rare. Damage was generally greater in the canopy than in the understorey, in particular for chewing and scraping damage, and the occurrence of mines. There was little difference in herbivory among differently managed forests and the effects of management on damage differed among regions, strata and damage types. Covariates such as wood volume, tree density and plant diversity weakly influenced herbivory, and effects differed between herbivory types. We conclude that despite of the relatively low number of species attacking beech; arthropod herbivory on beech is generally high. We further conclude that responses of herbivory to forest management are multifaceted and environmental factors such as forest structure variables affecting in particular microclimatic conditions are more likely to explain the variability in herbivory among beech forest plots.


Do neighborhood socioeconomic deprivation and low social cohesion predict coronary calcification?: the CARDIA study.
Growing evidence suggests that neighborhood characteristics may influence the risk of coronary heart disease. No studies have yet explored associations of neighborhood attributes with subclinical atherosclerosis in younger adult populations. Using data on 2,974 adults (1,699 women, 1,275 men) aged 32-50 years in 2000 from the Coronary Artery Disease Risk Development in Young Adults (CARDIA) Study and 2000 US Census block-group-level data, the authors estimated multivariable-adjusted associations of neighborhood socioeconomic deprivation and perceived neighborhood cohesion with odds of coronary artery calcification (CAC) 5 years later. Among women, the quartiles of highest neighborhood deprivation and lowest cohesion were associated with higher odds of CAC after adjustment for individual-level demographic and socioeconomic factors (for deprivation, odds ratio = 2.49, 95% confidence interval: 1.22, 5.08 (P for trend = 0.03); for cohesion, odds ratio = 1.87, 95% confidence interval: 1.10, 3.16 (P for trend = 0.02)). Associations changed only slightly after adjustment for behavioral, psychosocial, and biologic factors. Among men, neither neighborhood deprivation nor cohesion was related to CAC. However, among men in deprived neighborhoods, low cohesion predicted higher CAC odds (for interaction between neighborhood deprivation and cohesion, P = 0.03). This study provides evidence on associations of neighborhood deprivation and cohesion with CAC in younger, asymptomatic adults. Neighborhood attributes may contribute to subclinical atherosclerosis.


Preparation, characterization and photocatalytic behavior of WO3-fullerene/TiO2 catalysts under visible light.
WO3-treated fullerene/TiO2 composites (WO3-fullerene/TiO2) were prepared using a sol-gel method. The composite obtained was characterized by BET surface area measurements, X-ray diffraction, scanning electron microscopy, energy dispersive X-ray analysis, transmission electron microscopy, and UV-vis analysis. A methyl orange (MO) solution under visible light irradiation was used to determine the photocatalytic activity. Excellent photocatalytic degradation of a MO solution was observed using the WO3-fullerene, fullerene-TiO2, and WO3-fullerene/TiO2 composites under visible light. An increase in photocatalytic activity was observed, and WO3-fullerene/TiO2 has the best photocatalytic activity; it may attribute to the increase of the photo-absorption effect by the fullerene and the cooperative effect of the WO3.


Residual antibiotics in decontaminated human cardiovascular tissues intended for transplantation and risk of falsely negative microbiological analyses.
We investigated the presence of antibiotics in cryopreserved cardiovascular tissues and cryopreservation media, after tissue decontamination with antibiotic cocktails, and the impact of antibiotic residues on standard tissue bank microbiological analyses. Sixteen cardiovascular tissues were decontaminated with bank-prepared cocktails and cryopreserved by two different tissue banks according to their standard operating procedures. Before and after decontamination, samples underwent microbiological analysis by standard tissue bank methods. Cryopreserved samples were tested again with and without the removal of antibiotic residues using a RESEP tube, after thawing. Presence of antibiotics in tissue homogenates and processing liquids was determined by a modified agar diffusion test. All cryopreserved tissue homogenates and cryopreservation media induced important inhibition zones on both Staphylococcus aureus- and Pseudomonas aeruginosa-seeded plates, immediately after thawing and at the end of the sterility test. The RESEP tube treatment markedly reduced or totally eliminated the antimicrobial activity of tested tissues and media. Based on standard tissue bank analysis, 50% of tissues were found positive for bacteria and/or fungi, before decontamination and 2 out of 16 tested samples (13%) still contained microorganisms after decontamination. After thawing, none of the 16 cryopreserved samples resulted positive with direct inoculum method. When the same samples were tested after removal of antibiotic residues, 8 out of 16 (50%) were contaminated. Antibiotic residues present in tissue allografts and processing liquids after decontamination may mask microbial contamination during microbiological analysis performed with standard tissue bank methods, thus resulting in false negatives.


Reprogramming non-human primate somatic cells into functional neuronal cells by defined factors.
The common marmoset (Callithrix jacchus) is a New World primate sharing many similarities with humans. Recently developed technology for generating transgenic marmosets has opened new avenues for faithful recapitulation of human diseases, which could not be achieved in rodent models. However, the longer lifespan of common marmosets compared with rodents may result in an extended period for in vivo analysis of common marmoset disease models. Therefore, establishing rapid and efficient techniques for obtaining neuronal cells from transgenic individuals that enable in vitro analysis of molecular mechanisms underlying diseases are required. Recently, several groups have reported on methods, termed direct reprogramming, to generate neuronal cells by defined factors from somatic cells of various kinds of species, including mouse and human. The aim of the present study was to determine whether direct reprogramming technology was applicable to common marmosets. Common marmoset induced neuronal (cjiN) cells with neuronal morphology were generated from common marmoset embryonic skin fibroblasts (cjF) by overexpressing the neuronal transcription factors: ASCL1, BRN2, MYT1L and NEUROD1. Reverse transcription-polymerase chain reaction of cjiN cells showed upregulation of neuronal genes highly related to neuronal differentiation and function. The presence of neuronal marker proteins was also confirmed by immunocytochemistry. Electrical field stimulation to cjiN cells increased the intracellular calcium level, which was reversibly blocked by the voltage-gated sodium channel blocker, tetrodotoxin, indicating that these cells were functional. The neuronal function of these cells was further confirmed by electrophysiological analyses showing that action potentials could be elicited by membrane depolarization in current-clamp mode while both fast-activating and inactivating sodium currents and outward currents were observed in voltage-clamp mode. The 5-bromodeoxyuridine (BrdU) incorporation assay showed that cjiN cells were directly converted from cjFs without passing a proliferative state. Functional common marmoset neuronal cells can be obtained directly from embryonic fibroblasts by overexpressing four neuronal transcription factors under in vitro conditions. Overall, direct conversion technology on marmoset somatic cells provides the opportunity to analyze and screen phenotypes of genetically-modified common marmosets.


Treating constipation in palliative care: the impact of other factors aside from opioids.
Opioids are major contributing factors to the problem of constipation in palliative care. Whilst this is without doubt, it remains unclear how much other factors also contribute to the problem. The aim of this audit is to review what other contributing factors are present when methylnaltrexone, the peripheral opioid antagonist is prescribed for constipation. The medical records of people prescribed methylnaltrexone over a four-month period were reviewed to examine certain characteristics of people including the whether the reason for constipation was charted, whether other factors that could contribute to constipation were considered and the effectiveness of methylnaltrexone. Over the study period, 10 people received methylnaltrexone, only 4 of whom had a bowel action less than 24 hours after administration with 3 not having any bowel actions reported 6 days after administration. Whilst all were receiving opioids, the opioids doses were in the moderate range (61-200 mg morphine equivalent). However, all had other factors that could contribute to constipation including impaired functional status and medications with anti-cholinergic effects (mean anti-cholinergic load 4.5). In conclusion, methylnaltrexone is targeted treatment for the management of opioid-induced constipation. However, there is a percentage of people who fail to respond. The impact of other factors on the problem of constipation requires greater clarification.


An integrated framework for discovery and genotyping of genomic variants from high-throughput sequencing experiments.
Recent advances in high-throughput sequencing (HTS) technologies and computing capacity have produced unprecedented amounts of genomic data that have unraveled the genetics of phenotypic variability in several species. However, operating and integrating current software tools for data analysis still require important investments in highly skilled personnel. Developing accurate, efficient and user-friendly software packages for HTS data analysis will lead to a more rapid discovery of genomic elements relevant to medical, agricultural and industrial applications. We therefore developed Next-Generation Sequencing Eclipse Plug-in (NGSEP), a new software tool for integrated, efficient and user-friendly detection of single nucleotide variants (SNVs), indels and copy number variants (CNVs). NGSEP includes modules for read alignment, sorting, merging, functional annotation of variants, filtering and quality statistics. Analysis of sequencing experiments in yeast, rice and human samples shows that NGSEP has superior accuracy and efficiency, compared with currently available packages for variants detection. We also show that only a comprehensive and accurate identification of repeat regions and CNVs allows researchers to properly separate SNVs from differences between copies of repeat elements. We expect that NGSEP will become a strong support tool to empower the analysis of sequencing data in a wide range of research projects on different species.


The Graded Redefined Assessment of Strength Sensibility and Prehension: reliability and validity.
With the advent of new interventions targeted at both acute and chronic spinal cord injury (SCI), it is critical that techniques and protocols are developed that reliably evaluate changes in upper limb impairment/function. The Graded Redefined Assessment of Strength Sensibility and Prehension (GRASSP) protocol, which includes five subtests, is a quantitative clinical upper limb impairment measure designed for use in acute and chronic cervical SCI. The objectives of this study were to: (1) establish the inter-rater and test-retest reliability, and (2) establish the construct and concurrent validity with the International Standards of Neurological Classification of Spinal Cord Injury (ISNCSCI), Spinal Cord Independence Measure II (SCIM), and the Capabilities of Upper Extremity Questionnaire (CUE). The study protocol included repeated administration of the GRASSP to a cross-section of individuals with tetraplegia who were neurologically stable (n=72). ISNCSCI, CUE, and SCIM assessments were also administered. Two assessors examined the individuals over a 7-day period. Reliability was tested with intra-class correlation coefficients; construct validity was established with agreement/discordance analysis between the GRASSP and ISNCSCI sensory and motor items; and concurrent validity was tested with Spearman correlation coefficients. Inter-rater and test-retest reliability for all subtests within the GRASSP were above the hypothesized value of 0.80 (0.84-0.96 and 0.86-0.98, respectively). The GRASSP is about 50% more sensitive (construct validity) than the ISNCSCI when defining sensory and motor integrity of the upper limb; the subtests showed concurrence with the SCIM, SCIM self-care subscale, and CUE. The strongest concurrence to impairment was with self-perception of function (CUE) (0.57-0.83, p<0.0001). The GRASSP was found to demonstrate reliability, construct validity, and concurrent validity for use as a standardized upper limb impairment measure for individuals with tetraplegia.


Synthesis and thermal stability of perovskite alkali metal strontium borohydrides.
Three new perovskite-type bimetallic alkali metal strontium borohydride compounds, α-MSr(BH4)3 (M = K, Rb, Cs), have been synthesized and investigated by in situ synchrotron radiation powder X-ray diffraction, thermal analysis combined with mass spectrometry and Sievert's measurements. The bimetallic borohydrides were synthesized via an addition reaction between Sr(BH4)2 and MBH4 (M = K, Rb, Cs) by mechanochemical treatment. The Sr(BH4)2-NaBH4 system, which was treated in a similar manner, did not undergo reaction. All three α-MSr(BH4)3 compounds crystallize in the orthorhombic crystal system at room temperature: KSr(BH4)3 (P21cn), a = 7.8967(6), b = 8.2953(7), and c = 11.508(1) Å (V = 753.82(12) Å(3)). RbSr(BH4)3 (Pbn21), a = 8.0835(3), b = 8.3341(4), and c = 11.6600(5) Å (V = 785.52(6) Å(3)). CsSr(BH4)3 (P22121), a = 8.2068(9), b = 8.1793(9), and c = 6.0761(4) Å (V = 407.87(7) Å(3)). All three compounds are perovskite-type 3D framework structures built from distorted [Sr(BH4)6] octahedra. High-temperature polymorphs are identified to form at 258, 220 and 150 °C for MSr(BH4)3, M = K, Rb and Cs, respectively. The new compounds are thermally stable and decompose at T > 360 °C into SrB6, SrH2 and MBH4 (M = K, Rb, Cs).


Fixation locations when grasping partly occluded objects.
When grasping an object, subjects tend to look at the contact positions of the digits (A. M. Brouwer, V. H. Franz, D. Kerzel, & K. R. Gegenfurtner, 2005; R. S. Johansson, G. Westling, A. Bäckström, & J. R. Flanagan, 2001). However, these contact positions are not always visible due to occlusion. Subjects might look at occluded parts to determine the location of the contact positions based on extrapolated information. On the other hand, subjects might avoid looking at occluded parts since no object information can be gathered there. To find out where subjects fixate when grasping occluded objects, we let them grasp flat shapes with the index finger and thumb at predefined contact positions. Either the contact position of the thumb or the finger or both was occluded. In a control condition, a part of the object that does not involve the contact positions was occluded. The results showed that subjects did look at occluded object parts, suggesting that they used extrapolated object information for grasping. Additionally, they preferred to look in the direction of the index finger. When the contact position of the index finger was occluded, this tendency was inhibited. Thus, an occluder does not prevent fixations on occluded object parts, but it does affect fixation locations especially in conditions where the preferred fixation location is occluded.


Burnout among medical students interested in neurosurgery during the COVID-19 era.
The novel Coronavirus Disease 2019 (COVID-19) pandemic has posed unprecedented new stressors to medical student education. This national survey investigated the prevalence of burnout in U.S. medical students interested in pursuing neurosurgical residency during the COVID-19 pandemic. A 24-question survey was sent to all American Association of Neurological Surgeons (AANS) medical student chapter members. The abbreviated Maslach Burnout Inventory (aMBI) was used to measure the following burnout metrics: emotional exhaustion, depersonalization, and personal accomplishment. Bivariate analyses were conducted and multivariate analyses were performed using a logistic regression models. 254 medical students were included (response rate of 14.5%). The majority were male (55.1%), White (66.1%), and between their 2nd and 3rd years in medical school (62.6%). Burnout was identified in 38 (15.0%) respondents, a rate lower than reported in the pre-COVID era. In multivariate analysis, burnout was significantly associated with choosing not to pursue, or feeling uncertain about pursuing, a medical career again if given the choice (OR = 3.40, p = 0.0075), having second thoughts about choosing to pursue neurosurgery (OR = 3.47, p = 0.0025), attending a medical program in the Northeast compared to the Southeast (OR = 0.32, p = 0.027) or Southwest U.S. (OR = 0.30, p = 0.046), and indicating that one's future clinical performance will have worsened due to COVID-19 (OR = 2.71, p = 0.025). Our study demonstrates relatively low rates of burnout among U.S. medical students interested in pursuing neurosurgery during the COVID-19 pandemic. Our findings also demonstrate multiple factors may aid in early identification of burnout, highlighting potential opportunities for intervention.


Group for Research and Assessment of Psoriasis and Psoriatic Arthritis (GRAPPA): updated treatment recommendations for psoriatic arthritis 2021.
Since the second version of the Group for Research and Assessment of Psoriasis and Psoriatic Arthritis (GRAPPA) treatment recommendations were published in 2015, therapeutic options for psoriatic arthritis (PsA) have advanced considerably. This work reviews the literature since the previous recommendations (data published 2013-2020, including conference presentations between 2017 and 2020) and reports high-quality, evidence-based, domain-focused recommendations for medication selection in PsA developed by GRAPPA clinicians and patient research partners. The overarching principles for the management of adults with PsA were updated by consensus. Principles considering biosimilars and tapering of therapy were added, and the research agenda was revised. Literature searches covered treatments for the key domains of PsA: peripheral arthritis, axial disease, enthesitis, dactylitis, and skin and nail psoriasis; additional searches were performed for PsA-related conditions (uveitis and inflammatory bowel disease) and comorbidities. Individual subcommittees used a GRADE-informed approach, taking into account the quality of evidence for therapies, to generate recommendations for each of these domains, which were incorporated into an overall schema. Choice of therapy for an individual should ideally address all disease domains active in that patient, supporting shared decision-making. As safety issues often affect potential therapeutic choices, additional consideration was given to relevant comorbidities. These GRAPPA treatment recommendations provide up-to-date, evidence-based guidance on PsA management for clinicians and people with PsA.


It's Tea Time: Interference of Ayahuasca Brew on Discriminative Learning in Zebrafish.
Ayahuasca is a psychoactive brew traditionally used in shamanistic and vegetalistic rituals and has recently received lot of attention due to potential cognitive benefits. Ayahuasca effects are caused by the synergistic interaction of β-carbolines (harmine, harmaline and tetrahydroarmine) contained in Banisteriopsis caapi stalks combined with the N,N-dimethyltryptamine (DMT) from Psychotria viridis leaves, a potent agonist to serotonin (5-HT) receptors. The present study approaches the effects of chronic and acute exposure to two Ayahuasca concentrations (0.1 and 0.5 ml/L) on the cognitive ability to discriminate objects in a one-trial learning task in zebrafish. Based on the combination of concentrations and exposure regimens, we divided adult zebrafish in five treatment groups: acute 0.1 and 0.5 ml/L, chronic 0.1 and 0.5 ml/L, and control 0.0 (n = 20 for each group). Then we tested them in a memory task of object discrimination. Acute Ayahuasca exposed groups performed similarly to the control group, however chronically treated fish (13 days) presented both impaired discriminative performance and locomotor alterations. Overall, these results indicate that Ayahuasca is a potent psychoactive drug that, in chronic exposure, negatively affects mnemonic parameters in zebrafish. In single exposure it does not affects cognitive performance, but the higher concentration (0.5) affected locomotion. Moreover, we reinforce the importance of the zebrafish for behavioral pharmacological studies of drug screening, in special to psychedelic drug research.


Special issue on exercise immunology: current perspectives on aging, health and extreme performance.
The aim of this special issue is to highlight outstanding exemplars of empirical research and review papers that reflect the breadth of current developments in exercise immunology. The contributions to this issue are categorized according to four major themes: (1) exercise and immune-aging; (2) the effects of acute exercise on cellular shifts and gene expression; (3) the effects of exercise on immune regulation during stress and disease; and (4) extreme performance and the impact of dietary counter measures on immunity. We discuss the papers appearing in this issue, in accordance with these major themes, and summarize their important contributions to this exciting and expanding field.


Acute and long-term effects of exposure to sodium monofluoroacetate (1080) in sheep.
Acute and long-term effects of a single, relatively high oral dose (0.25 and 0.30 mg/kg) of sodium monofluoroacetate (1080) on the survival and productivity of sheep were evaluated to establish a better understanding of 1080 poisoning and identify more specific changes diagnostic of toxicosis. In survivors, clinical signs of acute 1080 toxicosis such as salivation and lethargy were generally very mild. Fasted animals were more prone to 1080 toxicity. In animals that died, more severe signs, including tachypnoea, dyspnoea, and tremors occurred for 15-20 min prior to death. 1080 concentrations were highest in the blood > heart > skeletal muscle > liver. 1080 could not be detected in any of these organs of the animals that survived. Serum citrate concentrations were elevated for 4 days after dosing. No clinical or biochemical abnormalities were found in any animal after 4 days. Histopathological lesions were most marked in the heart and lung with inflammation, necrosis, and scattered foci of fibrous tissue in the myocardium, pulmonary oedema and inflammation of the lung. No adverse long-term effects on general health or reproductive performance were observed in any sheep that survived the first 4 days following exposure to 1080. The most reliable diagnostic indicators of 1080 exposure in sheep were measurement of its residues in blood, skeletal muscle and ruminal contents, increased serum citrate concentration, elevated heart rate, and characteristic electrocardiograph changes (up to 4 days after exposure). Death from 1080 is most likely to occur within 96 h, and animals that survived this period appeared normal.


A systematic review of tools for predicting severe adverse events following patient discharge from intensive care units.
The discharge of patients from the intensive care unit (ICU) to a hospital ward is a common transition of care that is associated with error and adverse events. Risk stratification tools may help identify high-risk patients for targeted interventions, but it is unclear if proper tools have been developed. We searched Ovid EMBASE, Ovid MEDLINE, CINAHL, PUBMED and Cochrane Central Register of Controlled Trials from the earliest available date through March 2013, plus reference lists and citations of all studies included in the systematic review. Cohort studies were selected that described the derivation, validation or clinical impact of tools for predicting medical emergency team activation, ICU readmission or mortality following patient discharge from the ICU. Data were extracted on the study design, setting, population, sample size, tool (components, measurement properties) and outcomes. The literature search identified 9,926 citations, of which eight studies describing eight tools met the inclusion criteria. Reported outcomes included ICU readmission (n=4 studies), hospital mortality (n=3 studies) and both ICU readmission and hospital mortality (n=1 studies). Seven of the tools were comprised of distinct measurable component variables, while one tool used subjective scoring of patient risk by intensive care physicians. The areas under receiver operator curves were reported for all studies and ranged from 0.66 to 0.92. A single study provided a direct comparative analysis between two tools. We did not find any studies evaluating the impact of risk prediction on processes and outcomes of care. Eight risk stratification tools for predicting severe adverse events following patient discharge from ICU have been developed, but have undergone limited comparative evaluation. Although risk stratification tools may help clinician decision-making, further evaluation of the existing tools' effects on care is required prior to clinical implementation.


Exploring steric constraints on protein mutations using MAGE/PROBE.
When planning a mutation to test some hypothesis, one crucial question is whether the new side chain is compatible with the existing structure; only if it is compatible can the interpretation of mutational results be straightforward. This paper presents a simple way of using the sensitive geometry of all-atom contacts (including hydrogens) to answer that question. The interactive MAGE/PROBE system lets the biologist explore conformational space for the mutant side chain, with an interactively updated kinemage display of its all-atom contacts to the original structure. The Autobondrot function in PROBE systematically explores that same conformational space, outputting contact scores at each point, which are then contoured and displayed. These procedures are applied here in two types of test cases, with known mutant structures. In ricin A chain, the ability of a neighboring glutamate to rescue activity of an active-site mutant is modeled successfully. In T4 lysozyme, six mutations to Leu are analyzed within the wild-type background structure, and their Autobondrot score maps correctly predict whether or not their surroundings must shift significantly in the actual mutant structures; interactive examination of contacts for the conformations involved explains which clashes are relieved by the motions. These programs are easy to use, are available free for UNIX or Microsoft Windows operating systems, and should be of significant help in choosing good mutation experiments or in understanding puzzling results.


A regulatory role of LPCAT1 in the synthesis of inflammatory lipids, PAF and LPC, in the retina of diabetic mice.
Platelet-activating factor (PAF) and lysophosphatidylcholine (LPC) are potent inflammatory lipids. Elevated levels of PAF and LPC are associated with the onset of diabetic retinopathy and neurodegeneration. However, the molecular mechanisms underlying such defects remain elusive. LPCAT1 is a newly reported lysophospholipid acyltransferase implicated in the anti-inflammatory response by its role in conversion of LPC to PC. Intriguingly, the LPCAT1 enzyme also catalyzes the synthesis of PAF from lyso-PAF with use of acetyl-CoA as a substrate. The present studies investigated regulatory roles of LPCAT1 in the synthesis of inflammatory lipids during the onset of diabetes. Our work shows that LPCAT1 plays an important role in the inactivation of PAF by catalyzing the synthesis of alkyl-PC, an inactivated form of PAF with use of acyl-CoA and lyso-PAF as substrates. In support of a role of LPCAT1 in anti-inflammatory responses in diabetic retinopathy, LPCAT1 is most abundantly expressed in the retina. Moreover, LPCAT1 mRNA levels and acyltransferase activity toward lyso-PAF and LPC were significantly downregulated in retina and brain tissues in response to the onset of diabetes in Ins2(Akita) and db/db mice, mouse models of type 1 and type 2 diabetes, respectively. Conversely, treatment of db/db mice with rosiglitazone, an antidiabetes compound, significantly upregulated LPCAT1 mRNA levels concurrently with increased acyltransferase activity in the retina and brain. Collectively, these findings identified a novel regulatory role of LPCAT1 in catalyzing the inactivation of inflammatory lipids in the retina of diabetic mice.


Eggshell appearance does not signal maternal corticosterone exposure in Japanese quail: an experimental study with brown-spotted eggs.
Reproduction is a critical period for birds as they have to cope with many stressful events. One consequence of an acute exposure to stress is the release of corticosterone, the avian stress hormone. Prolonged stress can have negative impacts on the immune system, resulting in, for example, increased oxidative stress. Through maternal effects, females are known to modulate their investment in eggs content according to their own physiological condition. Less is known about maternal investment in eggshells, especially in pigments. The two main eggshell pigments may possess opposite antioxidant properties: protoporphyrin (brown) is a pro-oxidant, whereas biliverdin (blue-green) is an antioxidant. In Japanese quail, we know that the deposition of both pigments is related to female body condition. Thus, a chronic stress response may be reflected in eggshell coloration. Using female Japanese quails that lay brown-spotted eggs, we explored whether physiological exposure to corticosterone induces a change in female basal stress and antioxidant factors, and eggshell pigment concentration, spectrophotometric reflectance, and maculation coverage. We supplemented adult females over a 2 week period with either peanut oil (control) or corticosterone (treatment). We collected pre- and post-supplementation eggs and analysed the effect of corticosterone treatment on female physiology and eggshell appearance parameters. Except for corticosterone-fed birds which laid eggs with brighter spots, supplementation had no significant effect on female physiology or eggshell pigment concentration, reflectance and maculation. The change in eggshell spot brightness was not detected by a photoreceptor noise-limited color opponent model of avian visual perception. Our data confirms that eggshell reflectance in spotted eggs varies over the laying sequence, and spot reflectance may be a key factor that is affected by females CORT exposure, even if the changes are not detected by an avian visual model.


AIM inhibits apoptosis of T cells and NKT cells in Corynebacterium-induced granuloma formation in mice.
Apoptosis inhibitor expressed by macrophages (AIM) inhibits apoptosis of CD4(+)CD8(+) (CD4/CD8) double-positive thymocytes, and supports the viability of these cells on the thymic selection. However, pleiotropic functions of AIM have been suggested. In this study, heat-killed Corynebacterium parvum (C. parvum) was injected into mice carrying the homozygous mutation (AIM(-/-)) and wild-type (AIM(+/+)) mice, to investigate the role of AIM in the formation of hepatic granulomas. In AIM(-/-) mice, the size and the number of hepatic granulomas were larger, and the resorption of granulomas was more delayed than in AIM(+/+) mice. The production of interleukin-12 was more prominent in AIM(-/-) mice than in AIM(+/+) mice. In the liver of AIM(+/+) mice, expression of AIM messenger ribonucleic acid (mRNA) increased after C. parvum injection. In situ hybridization demonstrated that AIM mRNA was expressed in Kupffer cells and exudate macrophages in the liver, especially in granulomas. Larger numbers of T cells and natural killer T (NKT) cells underwent apoptosis in the granulomas of AIM(-/-) mice, suggesting that AIM prevents apoptosis of NKT cells and T cells in C. parvum-induced inflammation. Recombinant AIM (rAIM) protein significantly inhibited apoptosis of NKT cells and T cells obtained from C. parvum-stimulated livers in vitro. These results indicate that AIM functions to induce resistance to apoptosis within NKT cells and T cells, and supports the host defense in granulomatous inflammation.


Rest-task modulation of fMRI-derived global signal topography is mediated by transient coactivation patterns.
Recent resting-state functional MRI (fMRI) studies have revealed that the global signal (GS) exhibits a nonuniform spatial distribution across the gray matter. Whether this topography is informative remains largely unknown. We therefore tested rest-task modulation of GS topography by analyzing static GS correlation and dynamic coactivation patterns in a large sample of an fMRI dataset (n = 837) from the Human Connectome Project. The GS topography in the resting state and in seven different tasks was first measured by correlating the GS with the local time series (GSCORR). In the resting state, high GSCORR was observed mainly in the primary sensory and motor regions, whereas low GSCORR was seen in the association brain areas. This pattern changed during the seven tasks, with mainly decreased GSCORR in sensorimotor cortex. Importantly, this rest-task modulation of GSCORR could be traced to transient coactivation patterns at the peak period of GS (GS-peak). By comparing the topography of GSCORR and respiration effects, we observed that the topography of respiration mimicked the topography of GS in the resting state, whereas both differed during the task states; because of such partial dissociation, we assume that GSCORR could not be equated with a respiration effect. Finally, rest-task modulation of GS topography could not be exclusively explained by other sources of physiological noise. Together, we here demonstrate the informative nature of GS topography by showing its rest-task modulation, the underlying dynamic coactivation patterns, and its partial dissociation from respiration effects during task states.


Improving the care of patients with a hip fracture: a quality improvement report.
Hip fractures are associated with high rates of morbidity and mortality and their incidence is set to increase. The National Hip Fracture Database and the Best Practice Tariff were introduced with the aim of improving patient care. This complete audit cycle charts the substantial clinical improvements that were achieved in a busy district general hospital. The first audit cycle comprised 379 patients who were admitted between May 2012 and April 2013. The primary audit criterion was operative intervention within 36 h of admission. Variation according to the day of the week of admission was assessed to evaluate specific deficiencies in local service provision. The principle audit intervention was the introduction of two additional morning trauma lists. A re-audit of 162 patients was conducted prospectively between January 2014 and June 2014. Mean time to theatre was 49±39 h during the first audit cycle compared with 27±19 h (p<0.0001) during the second. Consequently, the proportion of patients undergoing operative intervention within 36 h of admission improved from 41% to 78% (p<0.0001). Overall achievement of Best Practice Tariff was significantly higher during the second cycle: 28% vs 73% (p<0.0001). Significant improvements in the quality of hip fracture care were achieved following this audit. These were accomplished by rigorously analysing the variation in Best Practice Tariff achievement according to the day of the week on which patients were admitted. Targeted interventions could therefore be introduced that addressed specific problems in local service provision.


A tissue-specific atlas of mouse protein phosphorylation and expression.
Although most tissues in an organism are genetically identical, the biochemistry of each is optimized to fulfill its unique physiological roles, with important consequences for human health and disease. Each tissue's unique physiology requires tightly regulated gene and protein expression coordinated by specialized, phosphorylation-dependent intracellular signaling. To better understand the role of phosphorylation in maintenance of physiological differences among tissues, we performed proteomic and phosphoproteomic characterizations of nine mouse tissues. We identified 12,039 proteins, including 6296 phosphoproteins harboring nearly 36,000 phosphorylation sites. Comparing protein abundances and phosphorylation levels revealed specialized, interconnected phosphorylation networks within each tissue while suggesting that many proteins are regulated by phosphorylation independently of their expression. Our data suggest that the "typical" phosphoprotein is widely expressed yet displays variable, often tissue-specific phosphorylation that tunes protein activity to the specific needs of each tissue. We offer this dataset as an online resource for the biological research community.


Absence of clonal beta and gamma T-cell receptor gene rearrangements in a subset of peripheral T-cell lymphomas.
The authors describe a set of seven peripheral T-cell lymphomas that lack detectable rearrangements of T-cell receptor (TCR) genes. All cases showed antigenic profiles consistent with T-cell lymphoma, including expression of Leu-5 (CD2) antigen. However, few other T-lineage markers were found, and none of the cases tested (6 of 7) bound antibody recognizing the constant region of the beta TCR protein. Each case showed exclusively germline configurations of DNA for the beta TCR genes in Southern blot analyses with the use of several different combinations of restriction enzymes and DNA hybridization probes. One case contained clonal rearrangements of the gamma TCR gene and of the immunoglobulin heavy chain gene. Our results suggest that certain cases of peripheral T-cell lymphoma may lack rearrangements of TCR genes--particularly those cases expressing restricted numbers of T-lineage antigens. In view of these findings, failure to detect rearrangements of TCR genes by Southern blot analyses is not necessarily inconsistent with malignant lymphocytic proliferations in T-lineage neoplasia.


Inducible nitric oxide synthase inhibitors of Chinese herbs. Part 2: naturally occurring furanocoumarins.
Inducible nitric oxide synthase (iNOS)-dependent production of nitric oxide (NO) plays an important role in inflammation. The effects of various naturally occurring furanocoumarins on NO production in lipopolysaccharide (LPS)-activated RAW 264.7 macrophage cells were evaluated in vitro. The results showed that angelicin, pimpinellin, sphondin, byakangelicol, oxypeucedanin, oxypeucedanin hydrate, xanthotoxin, and cnidilin are potential NO production inhibitors, and their IC50 values for inhibition of nitrite production were 19.5, 15.6, 9.8, 16.9, 16.8, 15.8, 16.6, and 17.7 microg/mL, respectively. Distinct structure-activity relationships were also revealed for the NO production inhibitory activities of these furanocoumarins. Activities of the angelicin type such as pimpinellin and sphondin were more potent than those of the psoralen type. Presence of a methoxy at the C6 position in the angelicin type seemed to be essential to augment the activity. Western blot analysis demonstrated that only sphondin dose-dependently inhibited the expression of the iNOS protein at 2.5-20 microg/mL. However, iNOS enzyme activity was stimulated with LPS for 12 h and sphondin was administered (20 microg/mL) for 24 h, which did not reasonably inhibit iNOS enzyme activity. L-NAME (100 microM), a known specific inhibitor of iNOS, was employed as a positive control with the same protocol and showed more than 50% inhibition activity. The results demonstrate that the NO production inhibitory activity of sphondin is due to the effect of iNOS expression, but not by direct inhibition of iNOS enzyme activity. Thus, sphondin may act as a potent inhibitor of NO production under tissue-damaging inflammatory conditions.


IBD risk loci are enriched in multigenic regulatory modules encompassing putative causative genes.
GWAS have identified >200 risk loci for Inflammatory Bowel Disease (IBD). The majority of disease associations are known to be driven by regulatory variants. To identify the putative causative genes that are perturbed by these variants, we generate a large transcriptome data set (nine disease-relevant cell types) and identify 23,650 cis-eQTL. We show that these are determined by ∼9720 regulatory modules, of which ∼3000 operate in multiple tissues and ∼970 on multiple genes. We identify regulatory modules that drive the disease association for 63 of the 200 risk loci, and show that these are enriched in multigenic modules. Based on these analyses, we resequence 45 of the corresponding 100 candidate genes in 6600 Crohn disease (CD) cases and 5500 controls, and show with burden tests that they include likely causative genes. Our analyses indicate that ≥10-fold larger sample sizes will be required to demonstrate the causality of individual genes using this approach.


Birth choices, the law, and medicine: balancing individual freedoms and protection of the public's health.
To many Americans, the idea of home birth, the use of a "direct-entry midwife," or both seem archaic. Although much of the professional medical community disapproves of either, state laws regarding birth choices vary dramatically and are not necessarily based on empirical findings of childbirth outcomes. Public health practitioners, policymakers, and consumers view childbirth from the perspectives of safety, cost, freedom of choice, quality of the care experience, and legality, yet the professional, policy, and lay literatures have not offered an unemotional, balanced presentation of evidence. Reviewing the full spectrum of literature from the United States and abroad, we present a Constitutional medical-legal analysis of whether home birth with direct-entry midwives is in fact a safe alternative to physician-attended hospital births, and whether there is a legal basis for allowing alternative health policy choices is such an important yet personal family matter as childbirth. The literature shows that low- to moderate-risk home births attended by direct-entry midwives are at least as safe as hospital births attended by either physicians or midwives. The policy ramifications include important changes in state regulation of medical and alternative health personnel, the allowance of the home as a medically acceptable and legal birth setting, and reimbursement of this lower-cost option through private and public health insurers.


Venlafaxine in treatment-resistant obsessive-compulsive disorder.
While selective serotonin reuptake inhibitors (SSRIs) are the first-line treatment of obsessive-compulsive disorder (OCD), approximately 40% of patients fail to respond to SSRIs. Venlafaxine is a serotonin-norepinephrine reuptake inhibitor (SNRI) that might be effective in the treatment of OCD, even among those who have failed previous SSRI trials. Thirty-nine patients who met DSM-IV criteria for OCD, including 29 who were resistant to prior SRI treatment trials, were treated with venlafaxine in an open, naturalistic fashion. Improvement was assessed using the Clinical Global Impressions-Improvement scale. Of 39 patients treated with venlafaxine, 27 (69.2%) were rated as sustained treatment responders. Of the 29 patients who did not respond to 1 or more previous SRI trials, 22 (75.9%) were rated as having sustained response to treatment. Mean dose of venlafaxine was 232.2 mg/day (range, 37.5-375 mg/day), and it was generally well tolerated. Venlafaxine may be beneficial to individuals with OCD, including those who have not responded to prior SSRI trials. However, these findings must be interpreted with caution, as the study is limited by its open, retrospective nature and its inclusion of patients with comorbid diagnoses and patients on concomitant medications. Prospective, controlled trials with a more homogeneous patient population are needed to replicate these preliminary findings.


KIT and mastocytosis.
KIT is a receptor tyrosine kinase that is functionally relevant for hematopoiesis, mast cell development and function, gametogenesis and melanogenesis. Normal KIT signaling requires binding to stem cell factor, and PI3K-Akt is one of the putative effector pathways. In humans, germline loss-of-function KIT mutations have been associated with piebaldism - an autosomal dominant condition characterized by depigmented patches of skin and hair. Gain-of-function KIT mutations are usually acquired and have been associated with myeloid malignancies including core binding factor acute myeloid leukemia and systemic mastocytosis (SM), germ cell tumors, gastrointestinal stromal tumors and sinonasal T cell lymphomas. KITD816V is the most prevalent KIT mutation in mast cell disease and occurs in more than 90% of the cases that fulfill the World Health Organization diagnostic criteria for SM. However, its precise pathogenetic contribution is not well understood. In clinical practice, SM is considered either indolent or aggressive depending on the respective absence or presence of symptomatic target organ dysfunction aside from skin disease. In general, conventional therapy for SM is suboptimal, and efforts are under way to develop and employ small molecule drugs that target mutant KIT.


Chemical analysis and amino acid content of temperature-sensitive mutants of Saccharomyces cerevisiae.
Temperature-sensitive (ts) mutants of Saccharomyces cerevisiae Y-196 were isolated by using UV irradiation and/or EMS treatment. They grew normally at permissive temperature (30 degrees C), but could not grow at restrictive temperature (38 degrees C). From chemical composition analysis, it was found that ts mutants AMY-159, and AMY-88 had the highest protein and RNA contents, respectively. When the cells were cultivated at 30 degrees C to stationary phase and then shifted to 38 degrees C for 4 hr, protein contents of wild-type and most of the ts mutants increased but RNA contents decreased. Glutamic acid, aspartic acid and lysine were the three major amino acids of protein in all isolates tested at both temperatures. When the ts mutants AMY-136, AMY-161, and AMY-157 were cultured at 38 degrees C for 4 hr, methionine contents increased Amino acid scores of ts mutants AMY-15 and AMY-88 increased when they were shifted to 38 degrees C for 2 to 4 doubling times. The amounts of amino acids produced in some ts mutants were higher than that of wild-type.