Endomucin, a CD34-like sialomucin, marks hematopoietic stem cells throughout development.
To detect as yet unidentified cell-surface molecules specific to hematopoietic stem cells (HSCs), a modified signal sequence trap was successfully applied to mouse bone marrow (BM) CD34(-)c-Kit(+)Sca-1(+)Lin(-) (CD34(-)KSL) HSCs. One of the identified molecules, Endomucin, is an endothelial sialomucin closely related to CD34. High-level expression of Endomucin was confined to the BM KSL HSCs and progenitor cells, and, importantly, long-term repopulating (LTR)-HSCs were exclusively present in the Endomucin(+)CD34(-)KSL population. Notably, in the yolk sac, Endomucin expression separated multipotential hematopoietic cells from committed erythroid progenitors in the cell fraction positive for CD41, an early embryonic hematopoietic marker. Furthermore, developing HSCs in the intraembryonic aorta-gonad-mesonephros (AGM) region were highly enriched in the CD45(-)CD41(+)Endomucin(+) fraction at day 10.5 of gestation (E10.5) and in the CD45(+)CD41(+)Endomucin(+) fraction at E11.5. Detailed analyses of these fractions uncovered drastic changes in their BM repopulating capacities as well as in vitro cytokine responsiveness within this narrow time frame. Our findings establish Endomucin as a novel cell-surface marker for LTR-HSCs throughout development and provide a powerful tool in understanding HSC ontogeny.


A review of histological outcomes from peri-menopausal and post-menopausal women with a cytological report of possible high grade abnormality: an alternative management strategy for these women.
To investigate the histological outcomes of women with cytological reports of possible high grade abnormality and to determine if patient age may be a clinically significant factor in determining the presence of high grade disease for this cytological category. Using annual quality reports supplied by the Queensland Pap Test Register, the histological outcomes for 2054 women with possible high grade cytological reports over a 4 year period were investigated to determine the clinical significance of this category in older aged women. Women in the total study group were aged between 16 and 84 years and outcomes were divided into women aged under 40 years and 40 years and over as well as those aged under 50 years and 50 years and over. Cytomorphological features were compared to determine if there were any significant differences between the outcome groups. The histological confirmation rate for high grade disease for all women was 53.1%. The incidence of histologically confirmed high grade disease was significantly higher in women aged under 40 years (59.7%) compared to women aged 40 years and over (40.3%) (p = 0.001). This was also true for women aged under 50 years compared to those aged 50 years and over (55.8% versus 37.5%) (p = 0.001). When the smears from these women were reviewed, a predominance of mature intermediate and superficial cells was evident in the background of the majority of slides in all histological outcome categories. Many of the smears in the negative outcome group showed reactive, reparative or inflammatory changes. The currently mandated Australian management guideline for women with a cytology report of possible high grade disease is referral for colposcopic and histological examination. Since the incidence of high risk human papillomavirus (HPV) decreases with age, a less invasive strategy may be to test women in these older age groups for high risk HPV before proceeding to colposcopy.


Observations on the correlation between posture and jaw position: a pilot study.
The aim of this pilot study was twofold. The first was to show a method for having an objective and dynamic analysis of body posture, evaluating weight distribution and its connections with different mandibular positions. The second was to verify if a neuromuscularly stimulated occlusal position, called myocentric occlusal position, is associated with a positive or negative postural charge. For the second aim a group of 20 subjects (including both males and females), was chosen. Posture of each subject was analyzed in three different conditions: centric occlusion, rest position and myocentric position. To evaluate the dynamic of posture a platform capable of measuring the weight on the feet supporting points and the related variations during time of observation and the swinging of body barycenter was used. The data showed that there is an improvement in the position of the barycenter from the centric occlusion to the myocentric position. Such an improvement can't be observed from the intercuspal position to the rest position. The results of this pilot study are discussed.


[Interaction between nutrition and production of IL-1 beta, TNF alpha, and IL-6 by peripheral blood monocytes in patients with lung cancer].
Altered nutrient intake and metabolism are responsible for the progressive loss of body weight observed in most patients with advanced cancer, but the precise mechanism is still controversial. Under stressful conditions, some inflammatory cytokines such as IL-1 beta, TNF alpha, and IL-6 have a hypermetabolic effect and cause proteolysis and lipolysis in muscle and in fat tissues. To elucidate the mechanism of malnutrition in patients with lung cancer and normal food intake, we focused on the relationship between abnormal metabolism and these inflammatory cytokines. Patients with lung cancer were confirmed to be malnourished, and this malnutrition was found to be caused by hypermetabolism as estimated with visceral proteins, plasma levels of amino acids, and anthropometric indices. The production of IL-1 beta, TNF alpha, and IL-6 by blood monocytes was significantly higher in these patients than in healthy controls, and it correlated significantly and inversely with indices of nutrition. The present results suggest that nutritional status and these cytokines are closely related in patients with lung cancer. IL-1 beta, TNF-alpha, and IL-6 may serve as anti-cancer bioactive molecules, but "overfunctioning" of these cytokines may induce a hypermetabolic status that causes malnutrition, i.e. cancer cachexia.


Two immunologically distinct forms of late-onset hypogammaglobulinaemia.
Most patients with late-onset bypogammaglobulinaemia have normal numbers of B cells. In addition, some of these patients have been shown to have circulating T cells which suppress immunolobulin production in vitro. From our present studies on a large series of patients it appears that these two findings distinguish two separate groups of patients. The majority of the patients have circulating B cells which are 'immature' in the sense that they produce IgM but very little IgG or IgA in vitro. These patients also fail to produce IgA in vivo but their T cells show normal function in vitro and normal T cell markers. Patients with the second form of the disease, including those with associated thymoma, have very few circulating B cells, and relatively preserved IgA production in vivo. In our series it was only this minority of patients whose T cells showed abnormal markers and increased suppressor activity in vitro.


Poly(ADP-ribose) polymerase activation mediates 1-methyl-4-phenyl-1, 2,3,6-tetrahydropyridine (MPTP)-induced parkinsonism.
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a neurotoxin that causes parkinsonism in humans and nonhuman animals, and its use has led to greater understanding of the pathogenesis of Parkinson's disease. However, its molecular targets have not been defined. We show that mice lacking the gene for poly(ADP-ribose) polymerase (PARP), which catalyzes the attachment of ADP ribose units from NAD to nuclear proteins after DNA damage, are dramatically spared from MPTP neurotoxicity. MPTP potently activates PARP exclusively in vulnerable dopamine containing neurons of the substantia nigra. MPTP elicits a novel pattern of poly(ADP-ribosyl)ation of nuclear proteins that completely depends on neuronally derived nitric oxide. Thus, NO, DNA damage, and PARP activation play a critical role in MPTP-induced parkinsonism and suggest that inhibitors of PARP may have protective benefit in the treatment of Parkinson's disease.


Negative interpersonal events mediate the relation between borderline features and aggressive behavior: findings from a nonclinical sample of undergraduate women.
Interpersonal dysfunction and aggression are features that are frequently found in individuals with borderline personality disorder (BPD); however, few studies have examined the possible causal relationship between aggressive actions and interpersonal problems. In a nonclinical sample of 98 women with a range of BPD features, the present study examined the prospective relationship between aggressive behaviors and negative interpersonal events using a weekly diary method. Results showed that higher BPD symptoms were related to higher aggression and more negative interpersonal events. Furthermore, the aggressive acts endorsed among women with more BPD features were more likely the effect, rather than the cause, of the negative interpersonal events they experienced. Implications for interventions targeting aggression among women with elevated BPD features and suggestions for future research are discussed.


Effects of donor-, pancreas-, and isolation-related variables on human islet isolation outcome: a systematic review.
Different factors have been reported to influence islet isolation outcome, but their importance varies between studies and are hampered by the small sample sizes in most studies. The purpose of this study was to perform a systematic review to assess the impact of donor-, pancreas-, and isolation-related variables on successful human islet isolation outcome. PubMed, Embase, and Web of Science were searched electronically in April 2009. All studies reporting on donor-, pancreas-, and isolation-related factors relating to prepurification and postpurification islet isolation yield and proportion of successful islet isolations were selected. Seventy-four retrospective studies had sufficient data and were included in the analyses. Higher pre- and postpurification islet yields and a higher proportion of successful islet isolations were obtained when pancreata were preserved with the two-layer method rather than University of Wisconsin solution in donors with shorter cold ischemia times (CITs) [1 h longer CIT resulted in an average decline of prepurification and postpurification yields and proportion of successful isolations of 59 islet equivalents (IEQs)/g, 54 IEQs/g, and 21%, respectively]. Higher prepurification yields and higher percentage of successful islet isolations were found in younger donors with higher body mass index. Lower yields were found in donation after brain death donors compared to donation after cardiac death donors. Higher postpurification yields were found for isolation with Serva collagenase. This review identified donor-, pancreas-, and isolation-related factors that influence islet isolation yield. Standardized reports of these factors in all future studies may improve the power and identify additional factors and thereby contribute to improving islet isolation yield.


Chronic neutrophilic leukemia: 2022 update on diagnosis, genomic landscape, prognosis, and management.
Chronic neutrophilic leukemia (CNL) is a rare, often aggressive myeloproliferative neoplasm (MPN) defined by persistent mature neutrophilic leukocytosis, bone marrow granulocyte hyperplasia, and frequent hepatosplenomegaly. The 2013 seminal discovery of oncogenic driver mutations in colony-stimulating factor 3 receptor (CSF3R) in the majority of patients with CNL not only established its molecular pathogenesis but provided a diagnostic biomarker and rationale for pharmacological targeting. In 2016, the World Health Organization (WHO) recognized activating CSF3R mutations as a central diagnostic feature of CNL. Other criteria include leukocytosis of ≥25 × 109 /L comprising >80% neutrophils with <10% circulating precursors and rare blasts, and absence of dysplasia or monocytosis, while not fulfilling criteria for other MPN. There is currently no standard of care for management of CNL, due in large part to the rarity of disease and dearth of formal clinical trials. Most commonly used therapeutic agents include conventional oral chemotherapy (e.g., hydroxyurea), interferon, and Janus kinase (JAK) inhibitors, while hematopoietic stem cell transplant remains the only potentially curative modality. Increasingly comprehensive genetic profiling in CNL, including new data on clonal evolution, has disclosed a complex genomic landscape with additional mutations and combinations thereof driving disease progression and drug resistance. Although accurate prognostic stratification and therapeutic decision-making remain challenging in CNL, emerging data on molecular biomarkers and the addition of newer agents, such as JAK inhibitors, to the therapeutic arsenal, are paving the way toward greater standardization and improvement of patient care.


Inhibition of Notch signalling has ability to alter the proximal and distal TCR signalling events in human CD3+ αβ T-cells.
The Notch signalling pathway is an important regulator of T cell function and is known to regulate the effector functions of T cells driven by T cell receptor (TCR). However, the mechanism integrating these pathways in human CD3+ αβ T cells is not well understood. The present study was carried out to investigate how Notch and TCR driven signalling are synchronized in human αβ T cells. Differential expression of Notch receptors, ligands, and target genes is observed on human αβ T cells which are upregulated on stimulation with α-CD3/CD28 mAb. Inhibition of Notch signalling by GSI-X inhibited the activation of T cells and affected proximal T cell signalling by regulating CD3-ζ chain expression. Inhibition of Notch signalling decreased the protein expression of CD3-ζ chain and induced expression of E3 ubiquitin ligase (GRAIL) in human αβ T cells. Apart from affecting proximal TCR signalling, Notch signalling also regulated the distal TCR signalling events. In the absence of Notch signalling, α-CD3/CD28 mAb induced activation and IFN-γ production by αβ T cells was down-modulated. The absence of Notch signalling in human αβ T cells inhibited proliferative responses despite strong signalling through TCR and IL-2 receptor. This study shows how Notch signalling cooperates with TCR signalling by regulating CD3-ζ chain expression to support proliferation and activation of human αβ T cells.


HPLC-based activity profiling of Salvia miltiorrhiza for MAO A and iNOS inhibitory activities.
In the course of a screening of plant extracts for potential CNS and anti-inflammatory activities, a dichloromethane extract of Salvia miltiorrhiza showed a pronounced inhibitory effect on recombinant monoamine oxidase A (MAO A) and on inducible NO synthase (iNOS) induction in Raw 267.4 cells. With the aid of HPLC-based profiling techniques, activities could be linked, to a significant extent, to peaks of tanshinone-type diterpenoids, four of which were subsequently isolated. The IC50 of the most active compound, 15,16-dihydrotanshinone I, on human recombinant MAO A was at 23 microM, and 2.4 microM on lipopolysaccharide-mediated iNOS induction in Raw 267.4 cells.


Smoking Status and Substance Use Treatment Outcomes Among Spanish Speakers Enrolled in Substance Abuse Treatment.
Smoking is highly prevalent among individuals with drug and alcohol disorders. Concurrent tobacco dependence treatment during substance use disorder (SUD) treatment is supported, yet the association between SUD treatment outcomes and smoking status has been understudied in minorities, including Latinos. Participants were 322 Spanish-speaking Latinos enrolled in a SUD treatment study in 5 U.S. cities. Logistic regression examined associations between baseline smoking status and treatment outcomes for nontobacco substance use at follow-up. Covariates included age, gender, level of education, marital status, treatment group, and mandated treatment status. Results indicated smokers had a reduced likelihood of abstinence for all nontobacco substances (p = .001) and their primary drug of use (p = .007). Findings contribute to the growing literature indicating a possible beneficial effect of smoking cessation services on SUD treatment, specifically among Latinos. Further research is needed to identify ideal smoking cessation treatments for Latinos.


Morphological effect of dichloromethane on alfalfa (Medicago sativa) cultivated in soil amended with fertilizer manures.
In this work, we investigated the morphological effect of dichloromethane (DCM) on alfalfa (Medicago sativa) plant. We studied in vitro the influence of its concentration on alfalfa germination. The plants were placed in pots for 15 weeks, and exposed to increasing concentrations of DCM (50 µg L-1 and 84 mg L-1). In addition, we examined the effect of two manures (cow and sheep), which were applied to a contaminated soil, on alfalfa plant growth. The effect of the presence of dichloromethane is obvious even in plant-soil manure system. In fact, in the event of contamination, the soil-cow manure mixture represents the best setting medium for the Alfalfa plant compared to other environments, regardless of the contamination level. Indeed, the presence of two types of manure does not allow the suppression of the inhibitory effect of dichloromethane on the mass of the dry matter of the aerial part which is 18.38% for the cow manure-amended soil and 13.96% for the sheep manure-amended soil.


Effects of magnetic resonance imaging on polymorphonuclear neutrophil adhesion.
Limited research has been performed concerning the effects of MR imaging on the immune system. In this study the influence of MR imaging exposure on polymorphonuclear neutrophil (PMN) adhesion was evaluated. In vivo and in vitro studies were performed in 10 patients undergoing an MR imaging procedure, PMN adhesion to a plastic surface, as well as the expression of adhesion molecules beta 2-integrins CD11b, CD18, and L-selectin on the surface of PMN were estimated. Exposure to MR imaging significantly increased adhesion of isolated PMNs to plastic surfaces. PMNs from blood samples obtained from patients undergoing MR imaging as well as from blood samples placed beside patients during MR imaging did not differ from controls in adhesion to plastic surfaces. Similarly, plasma from three tested samples did not change control PMN adhesion to plastic surface. Expression of beta 2-integrins (CD11, CD18) was significantly increased in samples left beside patients during MR imaging, while significantly decreased in samples obtained from patients after MR imaging exposure when compared to control samples. Expression of the surface adhesion molecule L-selectin on the surface of PMN decreased significantly in blood samples left beside patients during MR imaging. The results indicate that the PMN adhesion properties increase under the influence of MR imaging exposure. This phenomenon may be the result of direct stimulation of polymorphonuclear neutrophils by the exposure to MR imaging.


Use of rimabotulinum toxin for focal hypertonicity management in children with cerebral palsy with nonresponse to onabotulinum toxin.
The aim of this study was to review the effect of rimabotulinum toxin (BoNT-B) for focal hypertonicity management in children with cerebral palsy and secondary nonresponse to onabotulinum toxin treated at the authors' tertiary care academic medical center. A retrospective review of the medical treatment of children was conducted at the authors' institution (March 16, 2001, to August 2, 2002) using the key words botulinum toxin B and Myobloc (Solstice Neurosciences Inc, South San Francisco, CA). Demographic information was analyzed using descriptive statistics (number [percentage] and mean [range]). The Pearson χ test was used to evaluate differences in incidence of adverse events. Eighty-two children had BoNT-B injections (116 treatments). Overall, 26.8% (19/71) of the children or their parents/guardians reported no or minimal response to the injections, with 89.5% (17/19) of these children having secondary nonresponse to onabotulinum toxin. Adverse events were frequent but did not require hospitalization of any patient. No significant differences were found in incidence of adverse events related to BoNT-B dosing, medical fragility, or Gross Motor Function Classification System level. More than one-fourth of the children receiving BoNT-B injections had nonresponse, with most having previous nonresponse to onabotulinum toxin. Adverse events related to BoNT-B injections were frequent and unpredictable but not severe.


Regional white matter abnormalities in drug-naive, first-episode schizophrenia patients and their healthy unaffected siblings.
Shared neuropathological features between schizophrenia patients and their siblings may represent intermediate phenotypes of schizophrenia and can be used to investigate genetic susceptibility to the illness. This study aimed to discover regional white matter abnormalities in first-episode schizophrenia (FES) patients and their unaffected siblings compared to healthy subjects in the Chinese Han population using optimized Voxel-Based Morphometry (VBM). A total of 51 drug-naive, FES patients, 45 of their unaffected siblings and 59 healthy comparisons were studied with magnetic resonance imaging (MRI). FES patients exhibited significant regional white matter deficits in the left inferior frontal gyrus and left joint of external capsule and internal capsule compared with healthy subjects (corrected FDR, p<0.005). The sibling group also showed significant white matter deficits in these two regions compared with the healthy comparison group (uncorrected, p<0.001). White matter deficits with a less stringent threshold for significance in the left cerebellum anterior lobe, left middle frontal gyrus, left hippocampus, right anterior cingulate and right internal capsule were observed in patients compared to their siblings. Our findings extend those from previous VBM analyses showing that FES patients and their unaffected siblings may share white matter deficits in the left inferior frontal gyrus and the left joint of external capsule and internal capsule. These regional white matter deficits may be related to genetic factors related to schizophrenia susceptibility.


The microdynamics shaping the relationship between democracy and corruption.
Physics has a long tradition of laying rigorous quantitative foundations for social phenomena. Here, we up the ante for physics' forays into the territory of social sciences by (i) empirically documenting a tipping point in the relationship between democratic norms and corruption suppression, and then (ii) demonstrating how such a tipping point emerges from a micro-scale mechanistic model of spin dynamics in a complex network. Specifically, the tipping point in the relationship between democratic norms and corruption suppression is such that democratization has little effect on suppressing corruption below a critical threshold, but a large effect above the threshold. The micro-scale model of spin dynamics underpins this phenomenon by reinterpreting spins in terms of unbiased (i.e. altruistic) and biased (i.e. parochial) other-regarding behaviour, as well as the corresponding voting preferences. Under weak democratic norms, dense social connections of parochialists enable coercing enough opportunist voters to vote in favour of perpetuating parochial in-group bias. Society may, however, strengthen democratic norms in a rapid turn of events during which opportunists adopt altruism and vote to subdue bias. The emerging model outcome at the societal scale thus mirrors the data, implying that democracy either perpetuates or suppresses corruption depending on the prevailing democratic norms.


Patterns of gene expression in the developing adult sea urchin central nervous system reveal multiple domains and deep-seated neural pentamery.
The adult sea urchin central nervous system (CNS) is composed of five radial nerve cords connected to a circular nerve ring. Although much is known about the molecular mechanisms underlying the development and function of the nervous systems of many invertebrate and vertebrate species, virtually nothing is known about these processes in echinoderms. We have isolated a set of clones from a size-selected cDNA library prepared from the nervous system of the sea urchin Heliocidaris erythrogramma for use as probes. A total of 117 expressed sequence clones were used to search the GenBank database. Identified messages include genes that encode signaling proteins, cytoskeletal elements, cell surface proteins and receptors, cell proliferation and differentiation factors, transport and channel proteins, and a RNA DEAD box helicase. Expression was analyzed by RNA gel blot hybridization to document expression through development. Many of the genes have apparently neural limited expression and function, but some have been co-opted into new roles, notably associated with exocytotic events at fertilization. Localization of gene expression by whole-mount in situ hybridization shows that the morphologically simple sea urchin radial CNS exhibits complex organization into localized transcriptional domains. The transcription patterns reflect the morphological pentamery of the echinoderm CNS and provide no indication of an underlying functional bilateral symmetry in the CNS.


[Pancreatic trauma: 25-year experience].
The authors present 25 years experience in the treatment of traumatic injuries of pancreas. The clinical material covers 24 patients for the period 1984-2004. Twenty two (22) are gunshot injuries to the pancreas and 2 are blunt pancreatic injuries. From the traumatic lesions predominated the hematoma (I AAST grade) in 12 (50%) cases, lacerations (II AAST grade) in 9 (37,5%) cases, rupture of the pancreas in 3 (12,5%)--two patients with III grade and one patient with IV grade pancreatic injuries. Localizations of the lesions are as follow: body of the pancreas--13 (54,1%), tail--6 (25%), head--5 (20,8%) cases, Performed procedures are suture of the pancreatic capsule at 7 patients, distal pancreatic resection at 5 and laoparostomy at 4 patients. The following postoperative complications occurred: retroperitoneal phlegmon in 1 case, diffuse peritonitis--7, sepses--6, intraabdominal hemorrhage--4, hepato-renal syndrome--4, bronchopneumonia--4, stress ulcus--2, posttraumatic pancreatitis--1, pancreatic fistula--1. The mortality rate is 50% in the group of the gunshot injuries, while in cases of blunt trauma there were no dead. Conclusion--the authors point out the severity of the abdominal traumas in the presence of gunshot pancreatic injuries.


Light-Harvesting Complex II Adopts Different Quaternary Structures in Solution as Observed Using Small-Angle Scattering.
The high-resolution crystal structure of the trimeric major light-harvesting complex of photosystem II (LHCII) is often perceived as the basis for understanding its light-harvesting and photoprotective functions. However, the LHCII solution structure and its oligomerization or aggregation state may generally differ from the crystal structure and, moreover, also depend on its functional state. In this regard, small-angle scattering experiments provide the missing link by offering structural information in aqueous solution at physiological temperatures. Herein, we use small-angle scattering to investigate the solution structures of two different preparations of solubilized LHCII employing the nonionic detergents n-octyl-β-d-glucoside (OG) and n-dodecyl-β-D-maltoside (β-DM). The data reveal that the LHCII-OG complex is equivalent to the trimeric crystal structure. Remarkably, however, we observe─for the first time─a stable oligomer composed of three LHCII trimers in the case of the LHCII-β-DM preparation, implying additional pigment-pigment interactions. The latter complex is assumed to mimic trimer-trimer interactions which play an important role in the context of photoprotective nonphotochemical quenching.


Lower bioenergetic costs but similar immune responsiveness under a heat wave in urban compared to rural damselflies.
There is mounting evidence that the widespread phenotypic changes in response to urbanization may reflect adaptations caused by rapid evolutionary processes driven by urban-related stressors. Compared to increased habitat fragmentation and pollution, adaptations towards another typical urban-related stressor, that is higher and longer lasting very high temperatures (heat waves), are much less studied. Notably, the sensitivities to heat waves of life-history traits and important fitness-related physiological traits such as immune responsiveness and bioenergetic variables (energy availability, energy consumption and their balance) have never been contrasted between urban and rural populations. By conducting a laboratory common-garden experiment, we compared effects of a simulated heat wave on life history (survival and growth rate), immune responsiveness and bioenergetic variables between three urban and three rural populations of the damselfly Coenagrion puella. Because energy-mediated trade-off patterns may only be detected under energetically costly manipulations, all larvae were immune-challenged by simulating ectoparasitism by water mites. As expected, the simulated heat wave caused negative effects on nearly all response variables. The immune responsiveness, on the other hand, increased under the heat wave, consistent with a trade-off pattern between immune function and growth, and this similarly between urban and rural populations. A key finding was that urban larvae suffered less from the simulated heat wave compared to the rural larvae in terms of a lower heat wave-induced depletion in energy availability. This suggests an adaptation of urban populations to better cope with the stronger and more frequent heat waves in cities. Notably, this urbanization-driven evolution in the bioenergetic variables was not apparent in the absence of a heat wave. Given that changes in energy budgets have strong fitness consequences, our findings suggest that the evolved higher ability to cope with heat waves is fundamental for the survival of urban damselfly populations.


Pigments and proteins in green bacterial chlorosomes studied by matrix-assisted laser desorption ionization mass spectrometry.
We have used matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for mass determination of pigments and proteins in chlorosomes, the light-harvesting organelles from the photosynthetic green sulfur bacterium Chlorobium tepidum. By applying a small volume (1 microL) of a concentrated suspension of isolated chlorosomes directly to the target of the mass spectrometer we have been able to detect bacteriochlorophyll a and all the major homologs of bacteriochlorophyll c. The peak heights of the different bacteriochlorophyll c homologs in the MALDI spectra were proportional to peak areas obtained from HPLC analysis of the same sample. The same result was also obtained when whole cells of Chl. tepidum were applied to the target, indicating that MALDI-MS can provide a rapid method for obtaining a semiquantitative determination or finger-print of the bacteriochlorophyll homologs in a small amount of green bacterial cells. In addition to information on pigments, the MALDI spectra also contained peaks from chlorosome proteins. Thus we have been able with high precision to confirm the molecular masses of the chlorosome proteins CsmA and CsmE which have been previously determined by conventional biochemical and genetic methods, and demonstrate the presence of truncated versions of CsmA and CsmB.


Similar alteration of motor unit recruitment strategies during the anticipation and experience of pain.
A motor unit consists of a motoneurone and the multiple muscle fibres that it innervates, and forms the final neural pathway that influences movement. Discharge of motor units is altered (decreased discharge rate and/or cessation of firing; and increased discharge rate and/or recruitment of new units) during matched-force contractions with pain. This is thought to be mediated by nociceptive (pain) input on motoneurones, as demonstrated in animal studies. It is also possible that motoneurone excitability is altered by pain related descending inputs, that these changes persist after noxious stimuli cease, and that direct nociceptive input is not necessary to induce pain related changes in movement. We aimed to determine whether anticipation of pain (descending pain related inputs without nociceptor discharge) alters motor unit discharge, and to observe motor unit discharge recovery after pain has ceased. Motor unit discharge was recorded with fine-wire electrodes in the quadriceps of 9 volunteers. Subjects matched isometric knee-extension force during anticipation of pain (anticipation: electrical shocks randomly applied over the infrapatellar fat-pad); pain (hypertonic saline injected into the fat-pad); and 3 intervening control conditions. Discharge rate of motor units decreased during pain (P<.001) and anticipation (P<.01) compared with control contractions. De-recruitment of 1 population of units and new recruitment of another population were observed during both anticipation and pain; some changes in motor unit recruitment persisted after pain ceased. This challenges the fundamental theory that pain-related changes in muscle activity result from direct nociceptor discharge, and provides a mechanism that may underlie long-term changes in movement/chronicity in some musculoskeletal conditions.


Effect of 2-hydroxyethyl-methacrylate (HEMA) on the phagocytic and respiratory burst activity of human neutrophils and monocytes.
Neutrophils and monocytes/macrophages (MØ), found in oral mucosa and gingival sulcus, phagocytose and kill bacteria using products produced during a respiratory burst. 2-Hydroxyethyl-methacrylate (HEMA) is a major component released from resin glass ionomer and dental adhesives. Hence, in pulp and gingiva, phagocytes can come into contact with unpolymerized HEMA monomers. The aim of this study was to examine the effects of exposure to HEMA on neutrophil and monocyte bactericidal function. Blood collected from five female volunteers was exposed in vitro to HEMA for 2 h and then phagocytosis, respiratory burst, and cellular integrity were measured using flow cytometry. Respiratory burst was quantified by measuring fluorescent rhodamine 123 generated via oxidation of dihydrorhodamine 123. Cellular membrane integrity was evaluated by staining with propidium iodide. The respiratory burst activity of the neutrophils was significantly decreased by exposure to 7.5 and 15 mM HEMA. No significant effect of HEMA was seen on the number of granulocytes or monocytes capable of performing respiratory burst. Furthermore, there was no significant effect of HEMA on the phagocytic activity of the monocytes or the granulocytes. In conclusion, HEMA did not affect the phagocytosis activity of neutrophils; however, the ability of the cells to kill internalized prey was significantly reduced.


A new class of far-red and near-infrared biological labels based on alloyed semiconductor quantum dots.
Ternary cadmium selenium telluride quantum dots (CdSe(1-x)Tex, x = mole fraction of tellurium) have been prepared for potential use as constant-size biolabels with tunable fluorescence emission in the far-red and near-infrared (650-850 nm) spectral range. In contrast to particle size tuning reported for binary dots, we show that molar composition can be used to tune the optical and electronic properties of alloyed semiconductor nanocrystals without changing the particle size. A surprising finding is a nonlinear relationship between the composition and the absorption/emission energies, leading to new properties not obtainable from the parent CdSe and CdTe binary systems. Coating the alloy cores with a higher band-gap material such as CdS improves the fluorescence efficiencies to about 40-60% at room temperature and allows the preparation of water-soluble and biocompatible alloyed dots at similar quantum yields. A cadmium-rich surface is found to improve mercapto ligand binding and the long-term stability of water-soluble dots.


Phase I Trial of the Multi-kinase Inhibitor Cabozantinib, a CYP3A4 Substrate, plus CYP3A4-Interacting Antiretroviral Therapy in People Living with HIV and Cancer (AMC-087).
To evaluate the safety, pharmacokinetics, and pharmacodynamic effects of cabozantinib, a CYP3A4 substrate, in people living with human immunodeficiency virus and cancer receiving antiretrovirals (ARV). Patients received a reduced dose of cabozantinib (20 mg orally daily) with strong CYP3A4 inhibitors (ARV ritonavir or non-ARV cobicistat, stratum A), or a standard 60 mg dose with ARVs that are CYP3A4 inducers (efavirenz or etravirine, stratum B) or noninteracting ARVs (stratum C). Initial dose escalation in stratum A and stratum B was performed on the basis of tolerability. 36 patients received cabozantinib plus ARVs, including 20 in stratum A, 9 in B, and 7 in C. The recommended initial cabozantinib doses for stratum A, B, and C were 20, 60, and 60 mg, respectively. Doses of 40 or 60 mg plus CYP3A4 inhibitors in stratum A and 100 mg plus CYP3A4 inducers in stratum B were associated with excessive toxicity, whereas 60 mg with noninteracting ARVs was not. The steady state minimal concentrations were lower at 20 mg in stratum A or 60 mg in stratum B compared with 60 mg in stratum C, while total exposure was only lower in 60 mg in stratum B compared with 60 mg in stratum C. Activity was observed in Kaposi sarcoma and an AXL-amplified sarcoma. Cabozantinib as a single agent should be initiated at 20 mg daily and 60 mg daily when taken concurrently with ARVs that are strong CYP3A4 inhibitors and inducers, respectively, with consideration for subsequent escalation per current cabozantinib guidelines. See related commentary by Eisenmann and Sparreboom, p. 4999.


STUDY OF DOSES TO HIPPOCAMPUS OF INTERVENTIONAL CARDIOLOGISTS AND THEIR IMPLICATION FOR DOSIMETRIC MONITORING.
Dosimetric monitoring and protection of the personnel involved into interventional procedures is one of the key issues nowadays, yet doses received by their brains were left without rigorous consideration so far. The paper is the fast track of the results of the pilot study of possible magnitude of operator's doses with particular focus on difference between doses in left and right hippocampi and their relation to effective doses of personnel using protective aprons. Monte Carlo simulation of irradiation in a typical interventional cardiology (IC) operation room shows that for standard C-arm angulations difference in doses between left and right hippocampi can be as large as 5-fold (depending on energy and projection), under certain conditions dose to left (most exposed) hippocampus can be up to two times higher than effective dose estimated by a common double dosimetry algorithm. This finding calls for closer attention to possible manifestations of health detriment associated with occupational irradiation of left hippocampus in course of IC practice.


Activity-Based Sensing and Theranostic Probes Based on Photoinduced Electron Transfer.
Fluorescent probes have become powerful tools in detection, imaging and disease diagnosis due to their high sensitivity, specificity, fast response, and technical simplicity. In the last decades, researchers have made remarkable progress in developing signaling mechanisms to design fluorescent probes such as photoinduced electron transfer (PET), intramolecular charge transfer (ICT), and fluorescence resonance energy transfer (FRET). Typical PET is composed of a multicomponent system in which a fluorophore (electron acceptor) is separately linked with a recognition group (electron donor) via a short spacer. PET probes normally feature a low fluorescence background and significant fluorescence enhancement in response to targets. Recent research revealed that PET probes have also been used as theranostic agents, whose fluorescence and toxicity can be simultaneously activated by cancer-specific parameters. In this Account, we highlight the recent advances of rational design and applications of PET probes, focusing primarily on studies from our research group. For example, different from the case of the traditional single-atom electron donor (O, S, N, Se, Te, etc.) in typical PET, we used more a electron-rich pyrrole ring to "switch off" the fluorescence of the fluorophore more efficiently through an "enhanced PET" effect which provided a lower background fluorescence and higher signal-to-noise ratio. Furthermore, normal PET represents the main principle behind the design of small molecule "off-on" fluorescent sensors. We developed new PET platform through intramolecular space folding (folding PET) to overcome the difficulty of designing PET enzyme-targeting probes. Therefore, based on typical PET and these new PET concepts, we, for instance, reported PET probes for the detection of Zn2+ without proton interference, a BODIPY-based d-PET probe for reporting local hydrophilicity within lysosomes, and an "enhanced PET" fluorescent probe for imaging HClO in cancer cells. We also developed COX-2-specific probe for identifying cancer cells and quantifying cancer-related events, and a KIAA1363-sensitive probe for tracking solid tumors in living mice. Furthermore, we first applied an aminopeptidase N (APN)-sensitive probe based on PET for cancer diagnosis and therapy. We anticipate that further development of PET fluorescent probes providing more sensitivity and selectivity to analytes of interest will be equipped with more functions and play indispensable roles in the studies of pathology, diagnostics, and cancer therapies.


[The cooperation system for drug analysis by computer network].
Generally, the drugs are analyzed by the forensic medicine department in the most of autopsy cases. The forensic medicine department cannot always perform toxicological analysis because a shortage of a verified personnel, appropriate instruments, and financial resources. In order to overcome these difficulties and lack of resources, we developed a computer network called ml-poison. This network consists of the staff of forensic medicine, hospitals, governmental agencies and research facilities of police departments, etc. This network offers immediate and valuable information and expertise to these inexperienced in dealing with cases of poisoning. In cases in which the toxicological analysis cannot be performed in a facility, the network will recommend a facility where the analysis can be performed. Up this point, the network order system has assisted in many cases of poisoning. Although the effectiveness of the network for toxicological analysis has been proven, we still must deal with severed difficult problems: 1. The number of facilities assisting network orders is limited. 2. Who will pay the expenses involved in the analysis. 3. How to maintain security of the system. 4. What agency will assume responsibility for the management of the system.


Interaction of high concentrations of riluzole with recombinant skeletal muscle sodium channels and adult-type nicotinic receptor channels.
Riluzole is a neuroprotective drug that modulates glutamergic transmission but also blocks the inactivated state of voltage-gated neuronal sodium channels at very low concentrations (about 0.1 microM). After nausea, the most common adverse effect of riluzole is asthenia, which could be due to a block of muscle sodium channels or acetylcholine receptor channels. Using the patch-clamp technique, we applied riluzole on recombinant voltage-gated skeletal muscle sodium and adult nicotinic acetylcholine receptor channels expressed in a mammalian cell line (HEK 293). Riluzole blocked the inactivated state of voltage-gated skeletal muscle sodium channels, shifting the midpoint of the steady-state inactivation curve to more negative potentials, but only in comparatively high concentrations (> or = 0.1 mM). At these concentrations, riluzole also caused an open-channel block at acetylcholine receptor channels. We conclude that riluzole has only a mild blocking effect on the inactivated state of voltage-gated skeletal muscle sodium channels and nicotinic acetylcholine receptor channels. As the plasma concentration of riluzole in amyotrophic lateral sclerosis (ALS) patients approximates 2 microM, it seems unlikely that asthenia is caused by a block of skeletal muscle sodium channels or acetylcholine receptor channels by riluzole.


Scanning electron microscopy evaluation of the effect of etching agents on human enamel surface.
Acid etching promotes microporosities on enamel surface, which provide a better bonding surface to adhesive materials. The purpose of this study was to comparatively analyze the microstructure of enamel surface after etching with 37% phosphoric acid or with two self-etching primers, Non-rinse conditioner (NRC) and Clearfil SE Bond (CSEB) using scanning electron microscopy. Thirty sound premolars were divided into 3 groups with ten teeth each: Group 1: the buccal surface was etched with 37% phosphoric acid for 15 seconds; Group 2: the buccal surface was etched with NRC for 20 seconds; Group 3: the buccal surface was etched with CSEB for 20 seconds. Teeth from Group 1 were rinsed with water; teeth from all groups were air-dried for 15 seconds. After that, all specimens were processed for scanning electron microscopy and analyzed in a Jeol 6100 SEM. The results showed deeper etching when the enamel surface was etched with 37% phosphoric acid, followed by NRC and CSEB. It is concluded that 37% phosphoric acid is still the best agent for a most effective enamel etching.


Revisiting the effect of PCR replication and sequencing depth on biodiversity metrics in environmental DNA metabarcoding.
Environmental DNA (eDNA) metabarcoding is an increasingly popular tool for measuring and cataloguing biodiversity. Because the environments and substrates in which DNA is preserved differ considerably, eDNA research often requires bespoke approaches to generating eDNA data. Here, we explore how two experimental choices in eDNA study design-the number of PCR replicates and the depth of sequencing of PCR replicates-influence the composition and consistency of taxa recovered from eDNA extracts. We perform 24 PCR replicates from each of six soil samples using two of the most common metabarcodes for Fungi and Viridiplantae (ITS1 and ITS2), and sequence each replicate to an average depth of ~84,000 reads. We find that PCR replicates are broadly consistent in composition and relative abundance of dominant taxa, but that low abundance taxa are often unique to one or a few PCR replicates. Taxa observed in one out of 24 PCR replicates make up 21-29% of the total taxa detected. We also observe that sequencing depth or rarefaction influences alpha diversity and beta diversity estimates. Read sampling depth influences local contribution to beta diversity, placement in ordinations, and beta dispersion in ordinations. Our results suggest that, because common taxa drive some alpha diversity estimates, few PCR replicates and low read sampling depths may be sufficient for many biological applications of eDNA metabarcoding. However, because rare taxa are recovered stochastically, eDNA metabarcoding may never fully recover the true amplifiable alpha diversity in an eDNA extract. Rare taxa drive PCR replicate outliers of alpha and beta diversity and lead to dispersion differences at different read sampling depths. We conclude that researchers should consider the complexity and unevenness of a community when choosing analytical approaches, read sampling depths, and filtering thresholds to arrive at stable estimates.


Effects of Temperature on H Secretion and Uptake by Excised Flexor Cells during Dark-Induced Closure of Samanea Leaflets.
Previous studies reveal that dark-induced closure of Samanea leaflets is accompanied by H(+) secretion from flexor motor cells. We now report that flexor tissue excised in the light, incubated in a weakly buffered bathing solution, and then darkened at different temperatures (18 degrees C-30 degrees C) acidified the medium (indicating net H(+) efflux) at all temperatures tested, but most rapidly at the highest temperature. However, pH changes reversed direction after 20 to 70 minutes; the lower the temperature, the later pH reversal occurred, and the lower the pH at reversal and after 45 minutes. These data provide a basis for the previously reported promotive effect of low temperature on dark-induced leaflet closure, assuming net H(+) and K(+) fluxes are opposite in direction. Net H(+) efflux at all temperatures tested was greater when the impermeant molecule iminodiacetate replaced small permeant anions in the bathing solution, suggesting that H(+) uptake is coupled to anion uptake, probably via a H(+)/anion symport system. When permeant anions were deficient, the amount of malate in the tissue increased, presumably by new synthesis. Malate synthesis would substitute for H(+)/anion uptake in charge balance and in providing H(+) for cytoplasmic pH regulation.


Maximum static inspiratory and expiratory pressures with different lung volumes.
Maximum pressures developed by the respiratory muscles can indicate the health of the respiratory system, help to determine maximum respiratory flow rates, and contribute to respiratory power development. Past measurements of maximum pressures have been found to be inadequate for inclusion in some exercise models involving respiration. Maximum inspiratory and expiratory airway pressures were measured over a range of lung volumes in 29 female and 19 male adults. A commercial bell spirometry system was programmed to occlude airflow at nine target lung volumes ranging from 10% to 90% of vital capacity. In women, maximum expiratory pressure increased with volume from 39 to 61 cmH2O and maximum inspiratory pressure decreased with volume from 66 to 28 cmH2O. In men, maximum expiratory pressure increased with volume from 63 to 97 cmH2O and maximum inspiratory pressure decreased with volume from 97 to 39 cmH2O. Equations describing pressures for both sexes are: Pe/Pmax = 0.1426 Ln( %VC) + 0.3402 R2 = 0.95 Pi/Pmax = 0.234 Ln(100 - %VC) - 0.0828 R2 = 0.96 These results were found to be consistent with values and trends obtained by other authors. Regression equations may be suitable for respiratory mechanics models.


The effect of immobilization on the types of collagen synthesized in periarticular connective tissue.
The distribution of collagen types was studied in control and immobilized periarticular connective tissue to test the hypothesis that inflammatory tissue is the basis of the contracture process. The hypothesis derives from the fact that Type III collagen is prominent in inflammatory processes. The content of Type I and Type III collagen was determined by standard differential salt precipitation of the pepsin solubilized collagen, followed by resolution of the subunits on CM-cellulose. Analysis of the peptide fragments produced by cyanogen bromide was performed and their distribution determined by SDS gel electrophoresis. The collagen of control and immobilized periarticular connective tissue proved to be entirely Type I. Type III was not present. The periarticular connective tissue contracture process, therefore, is different from the intraarticular process in which fibrofatty connective tissue proliferation is prominent.


[Evaluation of microbial contamination of linens in industrial laundry processes].
Laundering linens and protecting them from microbiological recontamination are critical issues for the hotel and food industries and especially for hospitals. This study was performed to evaluate a sample of industrial laundries in Sardinia (Italy), to assess their compliance with national hygienic and sanitary regulations, along the complete laundering process. Study results indicate that industrial laundering processes are effective and that better awareness of staff who handle laundered textiles is required to reduce the risk of recontamination.


DNA modifications and neurological disorders.
Mounting evidence has recently underscored the importance of DNA methylation in normal brain functions. DNA methylation machineries are responsible for dynamic regulation of methylation patterns in discrete brain regions. In addition to methylation of cytosines in genomic DNA (5-methylcytosine; 5mC), other forms of modified cytosines, such as 5-hydroxymethylcytosine, 5-formylcytosine, and 5-carboxylcytosine, can potentially act as epigenetic marks that regulate gene expression. Importantly, epigenetic modifications require cognate binding proteins to read and translate information into gene expression regulation. Abnormal or incorrect interpretation of DNA methylation patterns can cause devastating consequences, including mental illnesses and neurological disorders. Although DNA methylation was generally considered to be a stable epigenetic mark in post-mitotic cells, recent studies have revealed dynamic DNA modifications in neurons. Such reversibility of 5mC sheds light on potential mechanisms underlying some neurological disorders and suggests a new route to correct aberrant methylation patterns associated with these disorders.


Adolescents' beliefs about marijuana use: a comparison of regular users, past users and never/occasional users.
A questionnaire investigating adolescents' opinions and experiences regarding marijuana use was administered to 163 adolescents and young adults (96 boys and 67 girls) aged 13 to 20 (mean age = 16.8, s.d. = 1.5). Items referred to marijuana and other substances' dangerousness, representations regarding the positive and negative consequences of marijuana use. Responses were compared according to marijuana use status (classified into never/occasional use, current regular use and past regular use). Results show that adolescents' opinions differ according to their experience with marijuana use. Current regular users evaluate marijuana as less dangerous, but alcohol and heroin as more dangerous in comparison with never/occasional and past users. Current and past users are more likely to define marijuana as a medical drug and a plant used in agriculture, and less likely to define it as an illegal drug. Current and past users evaluate marijuana use as a way to cope with stress, to relax to a greater extent than do never/occasional users do. The latter attribute more negative consequences to marijuana use such as diminished driving ability and school performance and a pathway to hard drugs.


Combining rapid diagnostic tests and dried blood spot assays for point-of-care testing of human immunodeficiency virus, hepatitis B and hepatitis C infections in Burkina Faso, West Africa.
People screened for human immunodeficiency virus (HIV) using rapid diagnostic tests (RDTs) in Africa remain generally unaware of their status for hepatitis B (HBV) and hepatitis C (HCV) infections. We evaluated a two-step screening strategy in Burkina Faso, using both HIV RDTs and Dried Blood Spot (DBS) assays to confirm an HIV-positive test, and to test for HBV and HCV infections. HIV counselling and point-of-care testing were performed at a voluntary counselling and testing centre with HBV, HCV status and HIV confirmation using DBS specimens, being assessed at a central laboratory. Serological testing on plasma was used as the reference standard assay to control for the performance of DBS assays. Nineteen out of 218 participants included in the study were positive for HIV using RDTs. A fourth-generation HIV ELISA and immunoblot assays on DBS confirmed HIV status. Twenty-four out of 25 participants infected with HBV were found positive for hepatitis B surface antigen (HBsAg) using DBS. One sample with a low HBsAg concentration on plasma was not detected on DBS. Five participants tested positive for HCV antibodies were confirmed positive with an immunoblot assay using DBS specimens. Laboratory results were communicated within 7 days to participants with no loss to follow up of participants between the first and second post-test counselling sessions. In conclusion, DBS collection during HIV point-of-care testing enables screening and confirmation of HBV, HCV and HIV infections. Diagnosis using DBS may assist with implementation of national programmes for HBV, HCV and HIV screening and clinical care in middle- to low-income countries.


Serum immunoglobulin E response in sputum positive patients with pulmonary tuberculosis.
A marked elevation of serum IgE level (6939 +/- 5651 IU/ml) was observed in 20 untreated and drug sensitive patients with sputum AFB positive pulmonary tuberculosis. Of them 18 were freshly diagnosed and were totally untreated. Their mean serum IgE level was 6715 +/- 5545 IU/ml. The remaining two patients presented with a relapse of the disease but were sensitive to all anti-tuberculosis drugs. Their serum IgE levels before retreatment were 2828 and 15079 IU/ml respectively. In the 13 chronic drug resistant patients who never had sputum conversion, the serum IgE level (1116 +/- 1897 IU/ml) was significantly lower (P < 0.01). It is suggested that the high serum IgE response in freshly diagnosed untreated and drug sensitive patients with illness of short duration may be related to strong early tuberculin hypersensitivity reaction and protective immunity, whereas low serum IgE response in chronic drug resistant cases with high bacterial load for prolonged duration may be related to the generation of suppressor CD8 lymphocytes eventually leading to suppression of B cells committed to the synthesis of protective serum IgE.


Dilute Semiflexible Polymers with Attraction: Collapse, Folding and Aggregation.
We review the current state on the thermodynamic behavior and structural phases of self- and mutually-attractive dilute semiflexible polymers that undergo temperature-driven transitions. In extreme dilution, polymers may be considered isolated, and this single polymer undergoes a collapse or folding transition depending on the internal structure. This may go as far as to stable knot phases. Adding polymers results in aggregation, where structural motifs again depend on the internal structure. We discuss in detail the effect of semiflexibility on the collapse and aggregation transition and provide perspectives for interesting future investigations.


Solubilized derivatives of perylenetetracarboxylic dianhydride (PTCDA) adsorbed on highly oriented pyrolytic graphite.
The effect on 2D molecular crystallization caused by the addition of propylthioether side groups to the 3,4,9,10-perylenetetracarboxylic dianhydride (PTCDA) molecule is investigated using scanning tunneling microscopy (STM). The molecule was deposited from 1-phenyloctane onto highly oriented pyrolytic graphite (HOPG) and imaged at the liquid-solid interface. We observe a different structure to previously reported arrangements of PTCDA due to the presence of the propylthioether side groups which inhibits the formation of the herringbone phase. A model, supported by calculations based on density functional theory, is proposed in which molecules form rows stabilized by hydrogen bonding.


Snail associates with EGR-1 and SP-1 to upregulate transcriptional activation of p15INK4b.
Snail is a multifunctional transcriptional factor that has been described as a repressor in many different contexts. It is also proposed as an activator in a few cases relevant to tumor progression and cell-cycle arrest. This study investigated the detailed mechanisms by which Snail upregulates gene expression of the CDK inhibitor p15(INK4b) in HepG2 induced by the tumor promoter tetradecanoyl phorbol acetate (TPA). Using deletion mapping, the TPA-responsive element on the p15(INK4b) promoter was located between 77 and 228 bp upstream of the transcriptional initiation site, within which the putative binding regions of early growth response gene 1 (EGR-1) and stimulatory protein 1 (SP-1) were found. Gene expression of EGR-1, Snail and SP-1 can be induced by TPA within 0.5-6 h. In addition, basal levels of SP-1, but not of the other two transcriptional factors, were observed. Blockade of TPA-induced gene expression of Snail, EGR-1 or SP-1 suppressed activation of the p15-pro228 reporter plasmid harboring the TPA-responsive element. More detailed deletion mapping and site-directed mutagenesis further concluded that the overlapping EGR-1/SP-1-binding site was required for TPA-induced p15-pro228 activation. In an EMSA, a DNA-protein complex was elevated by TPA, which can be blocked by antibodies against EGR-1, SP-1 or Snail at 6 h. Immunoprecipitation/western blotting demonstrated that TPA could trigger the association of EGR-1 with Snail or SP-1. Furthermore, a double chromatin immunoprecipitation assay verified that EGR-1 could form a complex with Snail or SP-1 on the TPA-responsive element after treatment with TPA for 2-6 h. Finally, we demonstrated a novel Snail-target region which could be bound by Snail and was also required for TPA-induced p15-pro228 activation. In conclusion, Snail associates with EGR-1 and SP-1 to mediate TPA-induced transcriptional upregulation of p15(INK4b) in HepG2.


Percutaneous gastrostomy and gastroenterostomy: 2. Clinical experience.
This report describes the authors' initial experience with percutaneous gastrostomy (PG) and gastroenterostomy (PGE) in 40 patients. Indications for PG and PGE included alimentation (35 patients) and small bowel decompression (five). Seldinger technique with air distension of the stomach via a nasogastric tube (20 patients) is a simple method to insert small (7-9 French) and firm catheters; tube exchanges for larger and softer catheters often are necessary by this method (23 procedures in 17 patients). Coaxial trocar technique (19 patients) permits initial insertion of softer and often larger catheters (9-14 French feeding tubes), which are less likely to clog or require exchange; the intragastric balloon support method facilitates trocar insertion. Now preferred is a system that uses 18-gauge needle puncture and allows coaxial insertion of a final soft feeding tube at the initial procedure. Small bowel catheter positioning (PGE) (31 patients) was more common than gastrostomy alone (8 patients); "downhill puncture" toward the gastric antrum assists direct guide-wire cannulation of the duodenum via the gastric puncture (12 patients). Five complications occurred; two were major and included catheter dislodgement in one patient. Another patient, who had a pharyngeal tumor, suffered profound respiratory difficulty from premedication and nasogastric tube malposition; patients with head and neck tumors present particular problems with nasogastric tube passage and airway monitoring. Inability to pass a nasogastric tube does not preclude PG and PGE, as direct puncture of the stomach is feasible.


Sources of individual variation in problem-solving performance in urban great tits (Parus major): Exploring effects of metal pollution, urban disturbance and personality.
Despite growing research effort, we have a limited understanding of how urban disturbance factors affect cognitive traits, such as innovative problem-solving. We performed an initial assessment of how metal pollution and urbanization levels on territories are related to problem-solving performance in urban great tits (Parus major), by presenting an obstacle removal test at nest boxes in three urban nest box populations that are exposed to different levels of metal pollution. We predicted that problem-solving capacity might be reduced within the most polluted population due to pollution-related neurological impairments. On the other hand, we predicted that problem-solving might positively correlate with urban disturbance levels on territories, because some past research suggests that problem-solving promotes persistence in urbanized habitats. We also assessed relationships between exploratory personality type, behavioral patterns during tests, and problem-solving performance, and examined the repeatability and fitness correlates of problem-solving. We found no evidence that behavioral patterns or problem-solving performance were related to metal exposure or exploratory personality, or that problem-solving promotes reproductive success. However, birds on territories exposed to more urban disturbance, as quantified by proximity to paths and roads, were more likely to problem-solve. Moreover, an aggressive problem-solving approach negatively predicted problem-solving success, and behavioral patterns during tests and problem-solving success were repeatable. Thus, rather than indicating negative effects of pollution or urban disturbance on problem-solving performance, our study provides preliminary support for the hypothesis that urbanization favors innovative problem-solving, and suggests that problem-solving could be associated with a personality dimension independent of exploratory personality.


Risk association of fasting insulin resistance index in non-diabetic heart failure patients: A case-control study at Karachi, Pakistan.
Present work investigates the risk association of insulin resistance (IResistance) in Non-Diabetic Heart Failure (NDHF) patients. Eighty (n=80) NDHF patients and same numbers of healthy controls were included to investigate with anthropometric measures, fasting blood glucose level (FBGL), serum insulin (SI), FIRI and β-cells quantification was computed through HOMA-IR. Mean rank assessment of NDHF patients showed higher significant (p<0.0001) set of values in FBGL, SI, FIRI and HOMA-IR, when compared with controls. High (p<0.0001 & p<0.05) risk in NDHF patients was associated in SI status (OR=8.93-95% CI: 4.1-19.42) and also in HOMA-IR (OR=6.6-95% CI: 3.30-13.19), when compared for Pearson value based probability through Chi Square (χ2 Test) values estimates of probability, respectively. Area under the curve (AUC) of targeted NDHF patients showed higher set of estimation (FBGL-AUC =0.667, SI-AUC =0.763, FIRI-AUC=0.780 and HOMA-IR-AUC=0.776). Association of determinants through Pearson's (r) correlation was found significantly (p<0.0001) linked with HOMA-IR and FIRI. Regression coefficient shows that for every additional unit score in FBGL and SI can expect HOMA-IR to increase by an average of 0.883 (for FBGL) and 0.0368 (for SI), respectively. Findings concluded the association of IResistance with greater risk estimation in NDHF patients.


COVID-19 in patients with rheumatic disease in Hubei province, China: a multicentre retrospective observational study.
In the ongoing COVID-19 pandemic, the susceptibility of patients with rheumatic diseases to COVID-19 remains unclear. We aimed to investigate susceptibility to COVID-19 in patients with autoimmune rheumatic diseases during the ongoing COVID-19 pandemic. We did a multicentre retrospective study of patients with autoimmune rheumatic diseases in Hubei province, the epicentre of the COVID-19 outbreak in China. Patients with rheumatic diseases were contacted through an automated telephone-based survey to investigate their susceptibility to COVID-19. Data about COVID-19 exposure or diagnosis were collected. Families with a documented history of COVID-19 exposure, as defined by having at least one family member diagnosed with COVID-19, were followed up by medical professionals to obtain detailed information, including sex, age, smoking history, past medical history, use of medications, and information related to COVID-19. Between March 20 and March 30, 2020, 6228 patients with autoimmune rheumatic diseases were included in the study. The overall rate of COVID-19 in patients with an autoimmune rheumatic disease in our study population was 0·43% (27 of 6228 patients). We identified 42 families in which COVID-19 was diagnosed between Dec 20, 2019, and March 20, 2020, in either patients with a rheumatic disease or in a family member residing at the same physical address during the outbreak. Within these 42 families, COVID-19 was diagnosed in 27 (63%) of 43 patients with a rheumatic disease and in 28 (34%) of 83 of their family members with no rheumatic disease (adjusted odds ratio [OR] 2·68 [95% CI 1·14-6·27]; p=0·023). Patients with rheumatic disease who were taking hydroxychloroquine had a lower risk of COVID-19 infection than patients taking other disease-modifying anti-rheumatic drugs (OR 0·09 [95% CI 0·01-0·94]; p=0·044). Additionally, the risk of COVID-19 was increased with age (adjusted OR 1·04 [95%CI 1·01-1·06]; p=0·0081). Patients with autoimmune rheumatic disease might be more susceptible to COVID-19 infection than the general population. National Natural Science Foundation of China and the Tongji Hospital Clinical Research Flagship Program.


[Comparison of platelet activators used in slide platelet aggregation test].
The study was purposed to explore the suitable platelet activators to be used in slide platelet aggregation test. Experiments were as follows: (1) to detect the intensity and time in 15 healthy donors' platelet aggregation tests induced by cationic propyl gallate (c-PG) and the usual platelet activators: ADP, collagen, epinephrine, arachidonic acid and ristocentin, respectively; (2) to detect the time in platelet aggregation tests of 15 healthy donors induced by c-PG and the above usual platelet activators respectively after addition of PGI2, cAMP or EDTA; (3) to detect the time in 15 healthy donors' platelet aggregation tests induced by c-PG after addition of heparin; (4) to detect the intensity and time of platelet aggregation induced by c-PG at the platelet count of (240-15) x 10(9)/L, (5) to detect the time of platelet aggregation induced by c-PG in eight patients each of whom had taken 100 mg aspirin per day for five days. The results showed that (1) c-PG reduced the strongest intensity of platelet aggregation and the time taken was appropriate, (2) c-PG was the most effective activator to reveal the inhibitive effect on platelet by PGI2, cAMP or EDTA, (3) 0.5 - 3 U/ml heparin did not significantly change the platelet aggregation induced by c-PG, (4) 15 healthy donors' platelet aggregation induced by c-PG displayed clearly on the slide until the platelet count below 30 x 10(9)/L, (5) The platelet aggregation time induced by c-PG was significantly prolonged in eight patients who had taken aspirin. In conclusion, compared to the usual platelet activators, c-PG has remarkable potential advantages when used in slide platelet aggregation test.


Role of hybrid endovascular suite in improving outcomes of surgical ligation of coronary artery fistula.
Coronary artery fistulae are rare congenital or acquired coronary artery anomalies that can lead to significant cardiovascular morbidity and mortality. Surgical ligation has long been utilized in the treatment of these abnormalities. However, there is a high rate of recurrence due to incomplete closure of the fistulae, especially when multiple channels are present. Transcatheter techniques have become an acceptable alternative with good outcomes. Nevertheless, not all fistulae are amenable to the transcatheter approach and surgical repair is the treatment of choice. Intraoperative coronary angiography can improve the outcomes of surgery but has only sparingly been used due to the technical difficulties in a standard operating suite. Hybrid suites are becoming quite common these days with the emergence of procedures such as endovascular stent grafting and percutaneous valves. These suites have a complete imaging set up like a traditional catheterization laboratory and are also full operating suites. This case report discusses the use and potential benefits of performing intraoperative coronary angiography in a dedicated hybrid suite to help guide and ensure complete surgical closure of all fistulous connections.


Single Cell Analysis of Reversibility of the Cell Death Program in Ethanol-Treated Neuronal PC12 Cells.
Neurodegenerative diseases are generally characterized clinically by the selective loss of a distinct subset of neurons and a slow progressive course. Mounting evidence in vivo indicates that large numbers of neurons pass through a long period of injury and dysfunction before the actual death of the cells. Whether these dying neurons can be rescued and return to a normal, functional state is uncertain. In the present study, we explored the reversibility of the neuronal cell death pathway at various stages by monitoring the dynamics of single cells with high-resolution live-cell spinning disk confocal microscopy in an in vitro neuronal cell death model. We exposed differentiated neuronal PC12 cells to ethanol as our cell death model. Results showed that exposure to 5% ethanol for 24 h induced cell death in >70% of the cells. Ethanol treatment for 3 h already induced cellular changes and damage such as reactive oxygen species generation, elevation of intracellular Ca2+ level, phosphatidylserine exposure, nuclear shrinkage, DNA damage, mitochondrial fragmentation and membrane potential loss, and retraction of neurites. These phenomena are often associated with programmed cell death. Importantly, after removing ethanol and further culturing these damaged cells in fresh culture medium, cells recovered from all these cell injuries and generated new neurites. Moreover, results indicated that this recovery was not dependent on exogenous NGF and other growth factors in the cell culture medium. Overall, our results suggest that targeting dying neurons can be an effective therapeutic strategy in neurodegenerative diseases.


Extracellular microtubules. The origin, structure, and attachment of flagellar hairs in Fucus and Ascophyllum antherozoids.
Mastigonemes (Flimmer) from the sperm of Ascophyllum and Fucus were found to consist of a tripartite structure-a ca. 2000-A tapered basal region, a closed microtubular shaft, and a group of terminal filaments. Each of these regions appears to be constructed of globular subunits with a center-to-center distance of about 45 A. The mastigoneme microtubule is of smaller diameter (170-190 A) than cytoplasmic microtubules in these or other plant cells. During the initial stages of flagellar ontogeny, structures similar to mastigonemes (presumptive mastigonemes) are found within membrane-limited sacs in the cytoplasm or within the perinuclear space. Mastigonemes at this time are generally not found on the flagellar surface. Later, when the anterior flagellum acquires mastigonemes, the presumptive mastigonemes are absent from the cytoplasm. The regularity of attachment of mastigonemes to the flagellar surface suggests that specific attachment sites are constructed on the plasma membrane during flagellar ontogeny. No evidence for penetration of the mastigoneme through the plasma membrane was obtained. The origin and structure of mastigonemes are discussed in relation to reports of the origin and structure of other microtubular systems.


Novel reactions of one-electron oxidized radicals of selenomethionine in comparison with methionine.
Pulse radiolysis studies on hydroxyl (*OH) radical reactions of selenomethionine (SeM), a selenium analogue of methionine, were carried out, and the resultant transient radical cations and their subsequent reactions have been reported. At pH<3, the >Se*-OH radical adducts produced on reaction of SeM with *OH radical were converted to selenium centered radical cations (Se*+M), which react with another molecule of SeM to form dimer radical cation M(Se therefore Se)M+. At pH 7, the >Se*-OH radical adducts were converted to a monomer radical of the type (Se therefore N)M+ that acquires intramolecular stability through interaction with the lone pair of the N atom and this radical is denoted as SeM*+. SeM*+ decayed by first order kinetics, and the reduction potential of the couple SeM*+/SeM was determined to be 1.21+/-0.05 V vs NHE at pH 7. SeM*+ oxidized ABTS2- and TMPD with rate constants of (2.5+/-0.1)x10(8) and (6.1+/-0.2)x10(8) M(-1) s(-1), respectively, and reacted with hydroxide ion with a rate constant of (3.8+/-0.9)x10(5) M(-1) s(-1). SeM*+ reacts with molecular oxygen, and the rate constant for this reaction was determined to be (4.3+/-0.2)x10(8) M(-1) s(-1); similar reaction with methionine could not be observed experimentally. Like methionine radical cations, SeM*+ undergoes decarboxylation, although with lesser yield, to produce reducing 3-methyl-selenopropyl amino radicals (referred as alpha-amino radicals). The formation of these radicals was confirmed both by the estimation of the liberated CO2 and by one-electron reduction of MV2+, thionine, and PNAP. These results have been supported by quantum chemical calculations. Implications of these results in the biological role of SeM have also been briefly discussed.


Virulence Genetics of an Erwinia amylovora Putative Polysaccharide Transporter Family Member.
The Gram-negative enterobacterium Erwinia amylovora causes fire blight disease in apple and pear trees. Lipopolysaccharides and the exopolysaccharide amylovoran are essential E. amylovora virulence factors. We found that mutations in rfbX disrupted amylovoran production and virulence in apple fruits and tree shoots and that the deletion of yibD suppressed the rfbX mutant phenotype. The level of expression of yibD was about 10-fold higher in the ΔrfbX mutant than the wild type. A forward genetic suppressor screen in the ΔrfbX mutant uncovered multiple mutations in yibD and supported the conclusion that the virulence defect of rfbX mutants is due to reduced amylovoran production. The yibD and rfbX genes are expressed as a two-gene operon, yibD rfbX The rfbX gene encodes a previously uncharacterized putative polysaccharide subunit transporter, while yibD encodes a predicted glycosyltransferase. Mutation of rfbX did not have a detectable effect on lipopolysaccharide patterns; however, the overexpression of yibD in both the wild-type and ΔyibD ΔrfbX genetic backgrounds disrupted both amylovoran and lipopolysaccharide production. Additionally, the overexpression of yibD in the ΔyibD ΔrfbX mutant inhibited bacterial growth in amylovoran-inducing medium. This growth inhibition phenotype was used in a forward genetic suppressor screen and reverse-genetics tests to identify several genes involved in lipopolysaccharide production, which, when mutated, restored the ability of the ΔyibD ΔrfbX mutant overexpressing yibD to grow in amylovoran-inducing medium. Remarkably, all the lipopolysaccharide gene mutants tested were defective in lipopolysaccharide and amylovoran production. These results reveal a genetic connection between amylovoran and lipopolysaccharide production in E. amylovoraIMPORTANCE This study discovered previously unknown genetic connections between exopolysaccharide and lipopolysaccharide production in the fire blight pathogen Erwinia amylovora This represents a step forward in our understanding of the biology underlying the production of these two macromolecules. Fire blight is an economically important disease that impacts the production of apples and pears worldwide. Few fire blight control measures are available, and growers rely heavily on antibiotic applications at bloom time. Both exopolysaccharide and lipopolysaccharide are E. amylovora virulence factors. Our results indicate that the overexpression of the yibD gene in E. amylovora disrupts both lipopolysaccharide production and exopolysaccharide production. This effect could potentially be used as the basis for the development of an antivirulence treatment for the prevention of fire blight disease.


The effect of the glycolipoprotein extract (G-90) from earthworm Eisenia foetida on the wound healing process in alloxan-induced diabetic rats.
Diabetes is now regarded as a major public health problem. The number of patients is estimated to increase to over 439 million cases by 2030. One of the major health clinical problems in patients with diabetes patients is impaired wound healing. Diabetic foot ulcer is a major complication of diabetes mellitus in 12 to 25% of patients, which increases the risk of damage in the limbs or amputation. The earthworm Eisenia foetida glycolipoprotein (as known G-90) is a blend of macromolecules with some biological properties including mitogenicity, anticoagulation, fibrinolysis, bacteriostatic and antioxidatiaon. Given the biological properties of G-90, this study was conducted to investigate the effect of extract obtained from the homogenate of Eisenia foetida (G-90) on the wound healing process in alloxan-induced diabetic rats. The results of the present study revealed that treatment by using G-90 can speed up the wound healing process, which is exactly similar to the effect of D-panthenol treatment in rats. These findings also demonstrated that G-90 treatment decreases the risk of infection in the wound site compared with D-panthenol treatment. In addition, histological analysis indicated that a better extracellular matrix formation with increased fibroblast proliferation, neovascularization, collagen synthesis and early epithelial layer formation was observed in G-90 treated group. Therefore, the G-90 could be considered as a new wound healing agent introducing promising therapeutic approaches in both human and veterinary medicine. Copyright © 2016 John Wiley & Sons, Ltd.


Procalcitonin as a diagnostic biomarker of sepsis: A tertiary care centre experience.
Despite the advancement in diagnostic modalities of sepsis, it is still a leading cause of morbidity and mortality. Differentiation between sepsis and non-infectious disease states remains a diagnostic challenge. Procalcitonin (PCT) is useful for the diagnosis of sepsis but it varies in cut-off ranges at different clinical settings. The aim of this study was to correlate serum PCT levels with cultures and to evaluate the best cut-off values with high sensitivity and specificity for PCT. This prospective study included 305 patients from different medical wards; the patients were classified into group I: controls (n=46), group II: culture-negative sepsis (n=76) and group III: culture-positive sepsis (n=196). Mean p value <0.05 was considered significant. PCT levels were significantly higher in group II and group III as compared with group I. In group II, the best cut-off point for PCT was 1.3ng/ml with 87.30% sensitivity and 78.26% specificity (area under curve 0.86). In group III, the best cut-off value of 2.20ng/ml with 98.47% sensitivity and 89.13% specificity was found (AUC 0.96). Procalcitonin can accurately differentiate culture-negative and culture-positive sepsis from non-infectious diseases, thus making it a promising biomarker in diagnosis of bacterial sepsis.


Molecular Characteristics, Clinical Significance, and Cancer Immune Interactions of Angiogenesis-Associated Genes in Gastric Cancer.
Immunotherapy has evolved as a critical option to treat diverse cancers. The active response to immunotherapy relies on the unique interaction between cancer and the tumor microenvironment (TME). Angiogenesis is one of the hallmarks of cancer. However, the association between angiogenesis and clinical outcome, immune cell infiltration, and immunotherapy remains unknown in gastric cancer (GC). We systematically assessed 36 angiogenesis-associated genes (AAGs) and comprehensively identified the correlation between angiogenesis and transcriptional patterns, prognosis, and immune cell infiltration. The AAG_score was applied to quantify the angiogenesis subtypes of each patient. We then evaluated their values in prognostic prediction and therapeutic responses in GC. We discussed the mutations of AAGs in GC specimens from genetic levels and identified their expression patterns from TCGA and GEO cohorts. We determined two different molecular subtypes and observed that AAG mutations were related to patients' clinicopathological characteristics, prognosis, and infiltrating TME. Next, an AAG_score for predicting overall survival (OS) was established and its reliable predictive ability in GC patients was confirmed. Furthermore, we created a highly reliable nomogram to facilitate the clinical viability of the AAG_score. A low AAG_score, characterized by elevated microsatellite instability-high, mutation burden, and immune activation, demonstrated a superior OS. Additionally, the AAG_score was remarkedly correlated with the cancer stem cell index and drug susceptibility. Collectively, we identified a prognostic AAG signature for GC patients. This signature may contribute to clarifying the characteristics of TME and enable the exploration of more potent immunotherapy strategies.


Intraspinal schwannoma of the accessory nerve.
The second example of a schwannoma originating from the cervical portion of the accessory nerve is reported. The tumour was diagnosed by MRI and confirmed by surgery. The tumour was small and was located entirely within the cervical subarachnoid space without causing any detectable neurological deficit.


BAW temperature sensitivity and coupling in langanite.
One of the new materials belonging to the trigonal class 32, to which quartz belongs, is langanite (LGN, La3Ga5.5Nb0.5O14). High-quality LGN single crystals are now available, and, although similar in composition and structure to langasite (LGS, La3Ga5SiO14), LGN has smaller thermal expansion coefficients and comparable piezoelectric constants to LGS. These are desirable material properties for both SAW and BAW applications that require low frequency dependence on temperature. This paper examines in detail the LGN characteristics: phase velocity, temperature coefficient of frequency (TCF), electromechanical coupling coefficient, and power flow angle for both singly and doubly rotated plate cuts. Contour plots of these characteristics are constructed, revealing orientation regions where zero TCF and high coupling exist and suggesting potentially interesting cuts for practical BAW device design. Temperature compensated cut regions with coupling coefficients as high as 0.16 are predicted, which is twice the value for AT-cut quartz, along with a temperature compensated cut with cubic behavior around room temperature for one of the sets of material constants used. With such desirable properties, LGN is a promising candidate material for BAW applications requiring low temperature sensitivity with superior bandwidth characteristics due to its values of coupling coefficient larger than quartz. Several other orientations with low TCF and high coupling are also identified.


Systemic lipolysis promotes physiological fitness in Drosophila melanogaster.
Since interventions such as caloric restriction or fasting robustly promote lipid catabolism and improve aging-related phenotypical markers, we investigated the direct effect of increased lipid catabolism via overexpression of bmm (brummer, FBgn0036449), the major triglyceride hydrolase in Drosophila, on lifespan and physiological fitness. Comprehensive characterization was carried out using RNA-seq, lipidomics and metabolomics analysis. Global overexpression of bmm strongly promoted numerous markers of physiological fitness, including increased female fecundity, fertility maintenance, preserved locomotion activity, increased mitochondrial biogenesis and oxidative metabolism. Increased bmm robustly upregulated the heat shock protein 70 (Hsp70) family of proteins, which equipped the flies with higher resistance to heat, cold, and ER stress via improved proteostasis. Despite improved physiological fitness, bmm overexpression did not extend lifespan. Taken together, these data show that bmm overexpression has broad beneficial effects on physiological fitness, but these effects did not impact lifespan.


Reproductive development modulates gene expression and metabolite levels with possible feedback inhibition of artemisinin in Artemisia annua.
The relationship between the transition to budding and flowering in Artemisia annua and the production of the antimalarial sesquiterpene, artemisinin (AN), the dynamics of artemisinic metabolite changes, AN-related transcriptional changes, and plant and trichome developmental changes were measured. Maximum production of AN occurs during full flower stage within floral tissues, but that changes in the leafy bracts and nonbolt leaves as the plant shifts from budding to full flower. Expression levels of early pathway genes known to be involved in isopentenyl diphosphate and farnesyl diphosphate biosynthesis leading to AN were not immediately positively correlated with either AN or its precursors. However, we found that the later AN pathway genes, amorpha-4,11-diene synthase (ADS) and the cytochrome P450, CYP71AV1 (CYP), were more highly correlated with AN's immediate precursor, dihydroartemisinic acid, within all leaf tissues tested. In addition, leaf trichome formation throughout the developmental phases of the plant also appears to be more complex than originally thought. Trichome changes correlated closely with the levels of AN but not its precursors. Differences were observed in trichome densities that are dependent both on developmental stage (vegetative, budding, and flowering) and on position (upper and lower leaf tissue). AN levels declined significantly as plants matured, as did ADS and CYP transcripts. Spraying leaves with AN or artemisinic acid inhibited CYP transcription; artemisinic acid also inhibited ADS transcription. These data allow us to present a novel model for the differential control of AN biosynthesis as it relates to developmental stage and trichome maturation and collapse.


Circulating tumor cell-blood cell crosstalk: Biology and clinical relevance.
Circulating tumor cells (CTCs) are the seeds of distant metastasis, and the number of CTCs detected in the blood of cancer patients is associated with a worse prognosis. CTCs face critical challenges for their survival in circulation, such as anoikis, shearing forces, and immune surveillance. Thus, understanding the mechanisms and interactions of CTCs within the blood microenvironment is crucial for better understanding of metastatic progression and the development of novel treatment strategies. CTCs interact with different hematopoietic cells, such as platelets, red blood cells, neutrophils, macrophages, natural killer (NK) cells, lymphocytes, endothelial cells, and cancer-associated fibroblasts, which can affect CTC survival in blood. This interaction may take place either via direct cell-cell contact or through secreted molecules. Here, we review interactions of CTCs with blood cells and discuss the potential clinical relevance of these interactions as biomarkers or as targets for anti-metastatic therapies.


Vitamin D status in adolescents and young adults with HIV infection.
Vitamin D status affects immune function and thus may affect the progress of HIV infection. Our goals were to assess vitamin D intake and status in subjects with HIV infection and in matched control subjects and to determine whether HIV infection was associated with vitamin D insufficiency. Plasma 25-hydroxyvitamin D [25(OH)D] concentrations and vitamin D intake were measured in a cross-sectional study of members of the Reaching for Excellence in Adolescent Health (REACH) cohort. The subjects were aged 14-23 y; 74% were female, and 72% were black. Mean (+/-SE) vitamin D intake from food was 30% greater (P = 0.023) in HIV-positive subjects (295 +/- 18 IU/d; n = 237) than in HIV-negative subjects (227 +/- 26 IU/d; n = 121). The prevalence of vitamin D supplement use was 29% (104 of 358 subjects) and did not differ significantly by HIV status (P = 0.87). Mean plasma 25(OH)D did not differ significantly (P = 0.62) between the HIV-positive (20.3 +/- 1.1 nmol/L; n = 238) and HIV-negative (19.3 +/- 1.7 nmol/L; n = 121) subjects, nor was HIV status a significant predictor of plasma 25(OH)D when multiple regression analysis was used to adjust for other variables. The prevalence of vitamin D insufficiency [plasma 25(OH)D < or = 37.5 nmol/L] in the subjects was 87% (312 of 359 subjects). HIV infection did not influence vitamin D status. The prevalence of vitamin D insufficiency in both HIV-positive and HIV-negative REACH subjects was high, perhaps because these disadvantaged, largely urban youth have limited sun exposure.


Prevalence of hyperhidrosis among medical students.
To identify the prevalence of hyperhidrosis among medical students of Manaus, State of Amazonas, Brazil. We conducted an observational, transversal, survey which examined the prevalence of primary hyperhidrosis among medical students of the Federal University of Amazonas and its relation to body mass index (BMI) and stress. Students were weighed and interviewed. We used questionnaires with questions recommended by the International Hyperhidrosis Society to relate hyperhidrosis to the daily activities of each person. Results were given by calculating the prevalence ratios and confidence intervals. Among the 293 students examined, it was found that a total of 16 (5.5%) students had barely tolerable or intolerable excessive sweating, interfering with daily activities. None had known causes of hyperhidrosis and 50% had family history. In all suffering from the condition the disease was bilateral, the mainly affected locations being: hands (35.7%), legs (21.4%), axilla (17.9), face (10 7%), back (7.1%), chest (3.6%) and abdomen (3.6%). There was no predominance regarding gender, age or BMI. We found a positive relationship with BMI and observed a prevalence ratio of 2.48 higher in overweight students than in normal weight or underweight ones. The prevalence of primary hyperhidrosis among medical students of Manaus was 5.5%. There is a positive non-statistical relationship with overweight and obesity. It was further noted an observational relationship with stress.


Partial Inhibition of Calcineurin Activity by Rcn2 as a Potential Remedy for Vps13 Deficiency.
Regulation of calcineurin, a Ca2+/calmodulin-regulated phosphatase, is important for the nervous system, and its abnormal activity is associated with various pathologies, including neurodegenerative disorders. In yeast cells lacking the VPS13 gene (vps13Δ), a model of VPS13-linked neurological diseases, we recently demonstrated that calcineurin is activated, and its downregulation reduces the negative effects associated with vps13Δ mutation. Here, we show that overexpression of the RCN2 gene, which encodes a negative regulator of calcineurin, is beneficial for vps13Δ cells. We studied the molecular mechanism underlying this effect through site-directed mutagenesis of RCN2. The interaction of the resulting Rcn2 variants with a MAPK kinase, Slt2, and subunits of calcineurin was tested. We show that Rcn2 binds preferentially to Cmp2, one of two alternative catalytic subunits of calcineurin, and partially inhibits calcineurin. Rcn2 ability to bind to and reduce the activity of calcineurin was important for the suppression. The binding of Rcn2 to Cmp2 requires two motifs in Rcn2: the previously characterized C-terminal motif and a new N-terminal motif that was discovered in this study. Altogether, our findings can help to better understand calcineurin regulation and to develop new therapeutic strategies against neurodegenerative diseases based on modulation of the activity of selected calcineurin isoforms.


Growth temperature effects on thylakoid membrane lipid and protein content of pea chloroplasts.
The lipid composition and level of unsaturation of fatty acids has been determined for chloroplast thylakoid membranes isolated from Pisum sativum grown under cold (4 degrees /7 degrees C) or warm (14 degrees /17 degrees C) conditions. Both the relative amounts of lipid classes and degree of saturation were not greatly changed for the two growth conditions. In cold-grown plants, there was a slightly higher linolenic and lower linoleic acid content for the glycolipids monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), and sulfoquinovosyldiacylglycerol. In contrast to thylakoid membranes, a non-thylakoid leaf membrane fraction including the chloroplast envelope, had a higher overall level of fatty acid unsaturation in cold-grown plants due mainly to an increase in the linolenic acid content of MGDG, DGDG, phosphatidylglycerol, and phosphatidylcholine. The most clear cut change in the thylakoid membrane composition was the lipid to protein ratio which was higher in the cold-grown plants.


CD4+ T cell responses in Balb/c mice with food allergy induced by trinitrobenzene sulfonic acid and ovalbumin.
The rapid increase in atopic diseases is potentially linked to increased hapten exposure, however, the role of haptens in the pathogenesis of food allergy remains unknown. Further studies are required to elucidate the cluster of differentiation 4 positive (CD4+) T cell response to food allergy induced by haptens. Dendritic cells were primed by trinitrobenzene sulfonic acid (TNBS) as a hapten or ovalbumin (OVA) as a model antigen, in a cell culture model. BALB/c mice were sensitized using TNBS and/or OVA. Intestinal Th1/Th2 cell and ovalbumin specific CD4+ T cells proliferation, intestinal cytokines (interleukin‑4 and interferon‑γ) in CD4+ T cells were evaluated. TNBS increased the expression of T cell immunoglobulin and mucin domain‑4 and tumor necrosis factor ligand superfamily member 4 in dendritic cells. Skewed Th2 cell polarization, extensive expression of interleukin‑4, reduced expression of interferon‑γ and forkhead box protein P3 were elicited following concomitant exposure to TNBS and OVA, with reduced regulatory T cells in the mouse intestinal mucosa, whereas a Th1 response was detected when challenged by TNBS or OVA alone. This data suggests that TNBS, as a hapten, combined with food antigens may lead to a Th2 cell response in the intestinal mucosa.


4,7-bis(4-pyridylethynyl)-2,1,3-benzothiadiazole and its dipyridinium diperchlorate.
The title compound, C20H10N4S, and its dipyridinium salt, 4,4'-(2,1,3-benzodiazol-4,7-diyldiethynyl)dipyridinium diperchlorate, C20H12N4S2+*2ClO4-, display bond alternation in the 2,1,3-benzothiadiazole rings, which suggests their quinonoid character. The dipyridinium dication molecules stack along the a axis and form a dimer with short S.N interheteroatom contacts [3.146 (4) A] between the two 1,2,5-thiadiazole rings. The dimer is surrounded by the perchlorate anions with which it forms a large number of intermolecular N-H...O and C-H...O hydrogen bonds.