Delayed transition to new cell fates during cellular reprogramming.
In many embryos specification toward one cell fate can be diverted to a different cell fate through a reprogramming process. Understanding how that process works will reveal insights into the developmental regulatory logic that emerged from evolution. In the sea urchin embryo, cells at gastrulation were found to reprogram and replace missing cell types after surgical dissections of the embryo. Non-skeletogenic mesoderm (NSM) cells reprogrammed to replace missing skeletogenic mesoderm cells and animal caps reprogrammed to replace all endomesoderm. In both cases evidence of reprogramming onset was first observed at the early gastrula stage, even if the cells to be replaced were removed earlier in development. Once started however, the reprogramming occurred with compressed gene expression dynamics. The NSM did not require early contact with the skeletogenic cells to reprogram, but the animal cap cells gained the ability to reprogram early in gastrulation only after extended contact with the vegetal halves prior to that time. If the entire vegetal half was removed at early gastrula, the animal caps reprogrammed and replaced the vegetal half endomesoderm. If the animal caps carried morpholinos to either hox11/13b or foxA (endomesoderm specification genes), the isolated animal caps failed to reprogram. Together these data reveal that the emergence of a reprogramming capability occurs at early gastrulation in the sea urchin embryo and requires activation of early specification components of the target tissues.


[Surgical therapy of chronic pyothorax].
A series of 224 patients with chronic pyothorax were treated in our hospital from 1959 to 1985. This paper reported the cure rate, relapse rate and quality of life after operation. One of the 224 patients died from shock within 24 hours after operation. 197 patients cuts (91.6%) healed within one month after operation, but 63 patients had recurrence of pyothorax, abscess of thoracic wall or fistula within three years after operation. By re-operation and/or local drug treatment, 49 of the 63 patients healed, comparing the living activity before and after operation in 96 patients showed that the surgical intervention might give them some improvement.


Adaptation of Corynebacterium glutamicum to salt-stress conditions.
Corynebacterium glutamicum is one of the biotechnologically most important microorganisms because of its ability to enrich amino acids extracellularly. Hence, C. glutamicum requires effective adaptation strategies against both hypo- and hyperosmotic stress. We give a comprehensive and coherent outline about the quantitative dynamics of C. glutamicum during adaptation to hyperosmotic stress at the transcript and protein levels. The osmolyte carrier ProP, playing a pivotal role in hyperosmotic stress defence, exhibits the strongest up-regulation of all proteins. A conspicuously regulated group comprises proteins involved in lipid biosynthesis of the cell envelope. This is in accordance with our observation of a more viscous and stickier cell envelope, which is supported by the findings of an altered lipid composition. Together with our results, showing that several transporters were down-regulated, this membrane adaptation appears to be one of C. glutamicum's major protection strategies against hyperosmotic stress. In addition, we demonstrate that no oxidative stress and no iron limitation occur during salt stress contrary to former postulations. Ultimately, it is remarkable that various proteins with divergent mRNA-protein dynamics and regulation have been observed. This leads to the assumption that there are still unknown mechanisms in between the bacterial transcription, translation and post-translation and that these are waiting to be unravelled.


Live Attenuated Leishmania donovani Centrin Knock Out Parasites Generate Non-inferior Protective Immune Response in Aged Mice against Visceral Leishmaniasis.
Visceral leishmaniasis (VL) caused by the protozoan parasite Leishmania donovani causes severe disease. Age appears to be critical in determining the clinical outcome of VL and at present there is no effective vaccine available against VL for any age group. Previously, we showed that genetically modified live attenuated L. donovani parasites (LdCen-/-) induced a strong protective innate and adaptive immune response in young mice. In this study we analyzed LdCen-/- parasite mediated modulation of innate and adaptive immune response in aged mice (18 months) and compared to young (2 months) mice. Analysis of innate immune response in bone marrow derived dendritic cells (BMDCs) from both young and aged mice upon infection with LdCen-/- parasites, showed significant enhancement of innate effector responses, which consequently augmented CD4+ Th1 cell effector function compared to LdWT infected BMDCs in vitro. Similarly, parasitized splenic dendritic cells from LdCen-/- infected young and aged mice also revealed induction of proinflammatory cytokines (IL-12, IL-6, IFN-γ and TNF) and subsequent down regulation of anti-inflammatory cytokine (IL-10) genes compared to LdWT infected mice. We also evaluated in vivo protection of the LdCen-/- immunized young and aged mice against virulent L. donovani challenge. Immunization with LdCen-/- induced higher IgG2a antibodies, lymphoproliferative response, pro- and anti-inflammatory cytokine responses and stimulated splenocytes for heightened leishmanicidal activity associated with nitric oxide production in young and aged mice. Furthermore, upon virulent L. donovani challenge, LdCen-/- immunized mice from both age groups displayed multifunctional Th1-type CD4 and cytotoxic CD8 T cells correlating to a significantly reduced parasite burden in the spleen and liver compared to naïve mice. It is interesting to note that even though there was no difference in the LdCen-/- induced innate response in dendritic cells between aged and young mice; the adaptive response specifically in terms of T cell and B cell activation in aged animals was reduced compared to young mice which correlated with less protection in old mice compared to young mice. Taken together, LdCen-/- immunization induced a significant but diminished host protective response in aged mice after challenge with virulent L. donovani parasites compared to young mice.


Epidemiological study of salivary gland tumours.
Tumours located in the salivary glands form the most heterogeneous group in all human oncological pathology. They show various epidemiological, clinical and evolutionary characteristics which separate them from other neoplasms of the head and neck. In this paper, we have carried out a study on their epidemiological aspects, collecting 80 cases diagnosed in the ENT Service of the University Hospital Complex of Santiago over 17 years. The incidence was 1.22 cases per 100,000 inhabitants per year. The frequency was higher in males (58.75%) and in the 7th decade of age. A predominance was noticed in females under 40 years of age and in males over this age, but the differences were not statistically significant. The most frequent site was the parotid gland, and we could not find any case in the sublingual gland. In 52.5% of cases the tumour was benign, pleomorphic adenoma being the most prevalent. Among malignant tumours, the epidermoid carcinoma stood out in our series. The prevalence of benign tumours in females and of malignant tumours in males was clear, with significant differences. We compare our results with the data published in the literature.


System and methods for wide-field quantitative fluorescence imaging during neurosurgery.
We report an accurate, precise and sensitive method and system for quantitative fluorescence image-guided neurosurgery. With a low-noise, high-dynamic-range CMOS array, we perform rapid (integration times as low as 50 ms per wavelength) hyperspectral fluorescence and diffuse reflectance detection and apply a correction algorithm to compensate for the distorting effects of tissue absorption and scattering. Using this approach, we generated quantitative wide-field images of fluorescence in tissue-simulating phantoms for the fluorophore PpIX, having concentrations and optical absorption and scattering variations over clinically relevant ranges. The imaging system was tested in a rodent model of glioma, detecting quantitative levels down to 20 ng/ml. The resulting performance is a significant advance on existing wide-field quantitative imaging techniques, and provides performance comparable to a point-spectroscopy probe that has previously demonstrated significant potential for improved detection of malignant brain tumors during surgical resection.


Comparison of four commercial, automated antigen tests to detect SARS-CoV-2 variants of concern.
A versatile portfolio of diagnostic tests is essential for the containment of the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) pandemic. Besides nucleic acid-based test systems and point-of-care (POCT) antigen (Ag) tests, quantitative, laboratory-based nucleocapsid Ag tests for SARS-CoV-2 have recently been launched. Here, we evaluated four commercial Ag tests on automated platforms and one POCT to detect SARS-CoV-2. We evaluated PCR-positive (n = 107) and PCR-negative (n = 303) respiratory swabs from asymptomatic and symptomatic patients at the end of the second pandemic wave in Germany (February-March 2021) as well as clinical isolates EU1 (B.1.117), variant of concern (VOC) Alpha (B.1.1.7) or Beta (B.1.351), which had been expanded in a biosafety level 3 laboratory. The specificities of automated SARS-CoV-2 Ag tests ranged between 97.0 and 99.7% (Lumipulse G SARS-CoV-2 Ag (Fujirebio): 97.03%, Elecsys SARS-CoV-2 Ag (Roche Diagnostics): 97.69%; LIAISON® SARS-CoV-2 Ag (Diasorin) and SARS-CoV-2 Ag ELISA (Euroimmun): 99.67%). In this study cohort of hospitalized patients, the clinical sensitivities of tests were low, ranging from 17.76 to 52.34%, and analytical sensitivities ranged from 420,000 to 25,000,000 Geq/ml. In comparison, the detection limit of the Roche Rapid Ag Test (RAT) was 9,300,000 Geq/ml, detecting 23.58% of respiratory samples. Receiver-operating-characteristics (ROCs) and Youden's index analyses were performed to further characterize the assays' overall performance and determine optimal assay cutoffs for sensitivity and specificity. VOCs carrying up to four amino acid mutations in nucleocapsid were detected by all five assays with characteristics comparable to non-VOCs. In summary, automated, quantitative SARS-CoV-2 Ag tests show variable performance and are not necessarily superior to a standard POCT. The efficacy of any alternative testing strategies to complement nucleic acid-based assays must be carefully evaluated by independent laboratories prior to widespread implementation.


Association of interleukin-1β genetic polymorphisms with cognitive performance in elderly females without dementia.
Interleukin-1β (IL-1β) is considered to have a role in age-related cognitive decline. A recent study has shown that a promoter polymorphism of the IL-1β gene (rs16944) is associated with cognitive performance in elderly males without dementia. In this study, we examined whether polymorphisms of the IL-1β gene also influence cognitive functions in elderly females. Cognitive functions were assessed by the Wechsler adult intelligence scale-revised (WAIS-R) in 99 elderly (60 years) females without dementia. We selected five tagging polymorphisms from the IL-1β gene and examined the associations with the WAIS-R scores. Significant associations were found between verbal intelligence quotient (IQ) and the genotypes of rs1143634 and rs1143633 (P=0.0037 and P=0.010, respectively). No significant associations of rs16944 genotype were found with verbal or performance IQ. However, individuals homozygous for the G allele of rs16944 achieved higher scores in digit span compared with their counterpart, which is consistent with the previous findings in males. These results suggest that IL-1β gene variation may have a role in cognitive functions in aging females as well as males.


Anthropogenic warming has increased drought risk in California.
California is currently in the midst of a record-setting drought. The drought began in 2012 and now includes the lowest calendar-year and 12-mo precipitation, the highest annual temperature, and the most extreme drought indicators on record. The extremely warm and dry conditions have led to acute water shortages, groundwater overdraft, critically low streamflow, and enhanced wildfire risk. Analyzing historical climate observations from California, we find that precipitation deficits in California were more than twice as likely to yield drought years if they occurred when conditions were warm. We find that although there has not been a substantial change in the probability of either negative or moderately negative precipitation anomalies in recent decades, the occurrence of drought years has been greater in the past two decades than in the preceding century. In addition, the probability that precipitation deficits co-occur with warm conditions and the probability that precipitation deficits produce drought have both increased. Climate model experiments with and without anthropogenic forcings reveal that human activities have increased the probability that dry precipitation years are also warm. Further, a large ensemble of climate model realizations reveals that additional global warming over the next few decades is very likely to create ∼ 100% probability that any annual-scale dry period is also extremely warm. We therefore conclude that anthropogenic warming is increasing the probability of co-occurring warm-dry conditions like those that have created the acute human and ecosystem impacts associated with the "exceptional" 2012-2014 drought in California.


[CMACPAR an modified parallel neuro-controller for control processes].
CMACPAR is a Parallel Neurocontroller oriented to real time systems as for example Control Processes. Its characteristics are mainly a fast learning algorithm, a reduced number of calculations, great generalization capacity, local learning and intrinsic parallelism. This type of neurocontroller is used in real time applications required by refineries, hydroelectric centers, factories, etc. In this work we present the analysis and the parallel implementation of a modified scheme of the Cerebellar Model CMAC for the n-dimensional space projection using a mean granularity parallel neurocontroller. The proposed memory management allows for a significant memory reduction in training time and required memory size.


ROCK-isoform-specific polarization of macrophages associated with age-related macular degeneration.
Age is a major risk factor in age-related macular degeneration (AMD), but the underlying cause is unknown. We find increased Rho-associated kinase (ROCK) signaling and M2 characteristics in eyes of aged mice, revealing immune changes in aging. ROCK isoforms determine macrophage polarization into M1 and M2 subtypes. M2-like macrophages accumulated in AMD, but not in normal eyes, suggesting that these macrophages may be linked to macular degeneration. M2 macrophages injected into the mouse eye exacerbated choroidal neovascular lesions, while M1 macrophages ameliorated them, supporting a causal role for macrophage subtypes in AMD. Selective ROCK2 inhibition with a small molecule decreased M2-like macrophages and choroidal neovascularization. ROCK2 inhibition upregulated M1 markers without affecting macrophage recruitment, underlining the plasticity of these macrophages. These results reveal age-induced innate immune imbalance as underlying AMD pathogenesis. Targeting macrophage plasticity opens up new possibilities for more effective AMD treatment.


Al(CN)6(3-) and Al(NC)6(3-) trianions.
At this time the smallest trianions observed in the gas phase are fluorinated fullerenes and large organic ring systems with attached sulfonic acid groups. Considerably smaller trianions have been predicted to be sufficiently stable for observation in mass spectrometers, but have not yet been detected. Here two isomers of the aluminium cyanide trianion, Al(CN)(6)(3-) and Al(NC)(6)(3-), are studied using ab initio methods. These two isomers are predicted to be electronically stable and to show substantial barriers with respect to dissociation of CN(-) units. Thus, the investigated trianions hit a sweat-spot regarding the possibility of detection, as they are by far more robust with respect to dissociation than alkali halide trianions, while at the same time materials from which these trianions can at least in principle be formed are much more readily available than those needed for producing small covalently bound trianions.


Flubromazolam - Basic pharmacokinetic evaluation of a highly potent designer benzodiazepine.
Since their first appearance on the Internet in 2012, designer benzodiazepines established as an additional, quickly growing compound class among new psychoactive substances. Data regarding pharmacokinetic parameters, metabolism, and detectability for new compounds are limited or often not available. One of these compounds, flubromazolam (8-bromo-6-(2-fluorophenyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine), the triazolo-analogue of flubromazepam, has been offered on the Internet from 2014 on. The purpose of the present study was to assess the period of detectability in biological samples along with preliminary basic pharmacokinetic parameters of the designer benzodiazepine flubromazolam. To investigate these, one of the authors ingested a capsule containing 0.5 mg of the drug. Metabolism studies and suitability tests for the detection with immunochemical assays were performed with the samples obtained from the self-experiment and five authentic case samples. Flubromazolam and its mono-hydroxylated metabolite were detectable by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in urine for up to 6.5 and 8 days, respectively (lower limit of quantification (LLOQ) flubromazolam: 0.1 ng/mL). Peak serum concentrations were as low as 8 ng/mL (8 h post ingestion). Glucuronides were also detected. The terminal elimination half-life could be estimated in the range of 10-20 h. Immunochemical assays yielded negative results for serum samples and positive results for urine samples for up to five days post ingestion. The presented data demonstrate the detectability of a single uptake of 0.5 mg of flubromazolam in hair samples collected two weeks after drug uptake by LC-MS3 (cmax 0.6 pg/mg; LOD 0.01 pg/mg). The detected metabolites were in good agreement with those described in other studies. Copyright © 2017 John Wiley & Sons, Ltd.


Differentiation of an embryonic stem cell to hemogenic endothelium by defined factors: essential role of bone morphogenetic protein 4.
Current approaches to differentiate embryonic stem (ES) cells to hematopoietic precursors in vitro use either feeder cell, serum, conditioned culture medium or embryoid body, methods that cannot avoid undefined culture conditions, precluding analysis of the fate of individual cells. Here, we have developed a defined, serum-free and low cell-density differentiation program to generate endothelial and hematopoietic cells within 6 days from murine ES cells. Our novel approach identifies a set of factors that are necessary and sufficient to differentiate ES cells into definitive hematopoietic precursors, as documented by the time-lapse video microscopy of the stepwise differentiation processes from single progenitors. Moreover, this defined milieu revealed the essential role of bone morphogenetic protein 4 (BMP4) in determining the hematopoietic/endothelial fate and demonstrated that the hemogenic fate in mesoderm is determined as early as day 4 of our differentiation protocol. Our ability to directly convert ES cells to endothelial and hematopoietic precursors should have important utilities for studies of hematopoietic development and personalized medicine in the future.


Optical spectra of synechocystis and spinach photosystem II preparations at 1.7 K: identification of the D1-pheophytin energies and stark shifts.
We report and compare highly resolved, simultaneously recorded absorption and CD spectra of active Photosystem II (PSII) samples in the range 440-750 nm. From an appropriately scaled comparison of spinach membrane fragment (BBY) and PSII core spectra, we show that key features of the core spectrum are quantitatively represented in the BBY data. PSII from the cyanobacterium Synechocystis 6803 display spectral features in the Qy region of comparable width (50-70 cm(-1) fwhm) to those seen in plant PSII but the energies of the resolved features are distinctly different. A comparison of spectra taken of PSII poised in the S1QA and S2QA(-) redox states reveals electrochromic shifts largely attributable to the influence of QA(-) on Pheo(D1). This allows accurate determinations of the Pheo(D1) Qy absorption positions to be at 685.0 nm for spinach cores, 685.8 nm for BBY particles, and 683.0 nm for Synechocystis. These are discussed in terms of earlier reports of the Pheo(D1) energies in PSII. The Qx transition of Pheo(D1) undergoes a blue shift upon Q(A) reduction, and we place a lower limit of 80 cm(-1) on this shift in plant material. By comparing the magnitude of the Stark shifts of the Qx and Qy bands of Pheo(D1), the directions of the transition-induced dipole moment changes, Deltamu(x) and Deltamu(y), for this functionally important pigment could be determined, assuming normal magnitudes of the Deltamu's. Consequently, Deltamu(x) and Deltamu(y) are determined to be approximately orthogonal to the directions expected for these transitions. Low-fluence illumination experiments at 1.7 K resulted in very efficient formation of QA(-). This was accompanied by cyt b(559) oxidation in BBYs and carotenoid oxidation in cores. No chlorophyll oxidation was observed. Our data allow us to estimate the quantum efficiency of PSII at this temperature to be of the order 0.1-1. No Stark shift associated with the S1-to-S2 transition of the Mn cluster is evident in our samples. The similarity of Stark data in plants and Synechocystis points to minimal interactions of Pheo(D1) with nearby chloropyll pigments in active PSII preparations. This appears to be at variance with interpretations of experiments performed with inactive solubilized reaction-center preparations.


Barriers to innovation in continuing medical education.
Criteria for maintenance of certification (MOC) emphasize the importance of competencies such as communication, professionalism, systems-based care, and practice performance in addition to medical knowledge. Success of this new competency paradigm is dependent on physicians' willingness to engage in activities that focus on less traditional competencies. We undertook this analysis to determine whether physicians' preferences for CME are barriers to participation in innovative programs. A geographically stratified, random sample of 755 licensed, practicing physicians in the state of Oregon were surveyed regarding their preferences for type of CME offering and instructional method and plans to recertify. Three hundred seventy-six of 755 surveys were returned for +/-5% margin of error at 95% confidence level; 91% of respondents were board certified. Traditional types of CME offerings and instructional methods were preferred by the majority of physicians. Academic physicians were less likely than clinical physicians to prefer nontraditional types of CME offerings and instructional methods. Multiple regression analyses did not reveal any significant differences based on demography, practice location, or physician practice type. Physicians who participate in CME select educational opportunities that appeal to them. There is little attraction to competency-based educational activities despite their requirement for MOC. The apparent disparity between the instructional methods a learner prefers and those that are the most effective in changing physician behavior may represent a barrier to participating in more innovative CME offerings and instructional methods. These findings are important for medical educators and CME program planners developing programs that integrate studied and effective educational methods into CME programs that are attractive to physicians.


Sclerosing bone dysplasias: review and differentiation from other causes of osteosclerosis.
Sclerosing bone dysplasias are skeletal abnormalities of varying severity with a wide range of radiologic, clinical, and genetic features. Hereditary sclerosing bone dysplasias result from some disturbance in the pathways involved in osteoblast or osteoclast regulation, leading to abnormal accumulation of bone. Several genes have been discovered that, when disrupted, result in specific types of hereditary sclerosing bone dysplasia (osteopetrosis, pyknodysostosis, osteopoikilosis, osteopathia striata, progressive diaphyseal dysplasia, hereditary multiple diaphyseal sclerosis, hyperostosis corticalis generalisata), many of which exhibit similar pathologic mechanisms involving endochondral or intramembranous ossification and some of which share similar underlying genetic defects. Nonhereditary dysplasias include intramedullary osteosclerosis, melorheostosis, and overlap syndromes, whereas acquired syndromes with increased bone density, which may simulate sclerosing bone dysplasias, include osteoblastic metastases, Paget disease of bone, Erdheim-Chester disease, myelofibrosis, and sickle cell disease. Knowledge of the radiologic appearances, distribution, and associated clinical findings of hereditary and nonhereditary sclerosing bone dysplasias and acquired syndromes with increased bone density is crucial for accurate diagnosis.


The FAP motif within human ATG7, an autophagy-related E1-like enzyme, is essential for the E2-substrate reaction of LC3 lipidation.
ATG7 is an autophagy-related E1-like enzyme that is essential for two ubiquitination-like reactions, ATG12-conjugation and LC3-lipidation. The existence of functional sequences at the amino-terminal region of human ATG7 remains uncertain. Mutational analyses of ATG7 revealed that both mutant ATG7ΔFAP lacking the FAP motif and ATG7FAPtoDDD, in which the Phe15-Ala16-Pro17 sequence was changed to Asp-Asp-Asp, could not complement defects in endogenous ATG12-conjugation and LC3-lipidation when expressed in Atg7-deficient mouse embryonic fibroblasts (MEFs). However, wild-type ATG7 complemented the defects in these cells. Overexpression of GFP-ATG10 and GFP-ATG12 rescued a defect in ATG12-conjugation in Atg7-deficient MEFs expressing mutant ATG7ΔFAP and ATG7FAPtoDDD, whereas overexpression of all ATG proteins related to ATG12-conjugation and LC3-lipidation could not rescue a defect in LC3-lipidation in Atg7-deficient MEFs expressing these ATG7 mutants. Both ATG7ΔFAP and ATG7FAPtoDDD mutants showed severe defects in the formation of an E2-substrate intermediate of ATG3 with LC3 in LC3-lipidation, but were able to form an E1-substrate intermediate of ATG7 with LC3 and the E1- and E2-substrate intermediates in ATG12-conjugation with reduced efficiency. These ATG7 mutants could also form the ATG12-ATG3 conjugate. Co-immunoprecipitation experiments revealed that the FAP motif of ATG7 is essential for the interaction of ATG7 with ATG3, but not for ATG7-homodimerization. These results indicated that the FAP motif of ATG7 is indispensable for formation of the ATG3-LC3 E2-substrate intermediate through the interaction of ATG7 with ATG3.


[The effect of hemosorption on the activity of the kallikrein-kinin system in patients with hemorrhagic pancreatitis].
The kallikrein-kinin system (KKS) is activated in all forms of acute pancreatitis. The severity of toxemia in patients with pancreatitis correlates with KKS activity. 2-3 hemosorption procedures performed in patients with hemorrhagic pancreatitis in the first 1, 2 and 3, 4 days normalizes KKS activity by days 3-5, leads to a rapid improvement of the clinical manifestations of the disease, shortens the treatment period and improves the results.


[Diagnostic guidelines for complex regional pain syndrome].
The complex regional pain syndrome, a special form of neuropathic pain, develops after a minor trauma of the distal limbs. Besides the presentation of pain disproportional to the inciting event, further plus and minus symptoms in the form of sensory, vasomotor, sudomotor-oedematous and motor-trophic changes can be found. Interindividually and intraindividually, occurrence of these symptoms differs widely and single symptoms can be lacking completely. A gold standard in diagnosing CRPS has not been found yet, diagnostics are based on the patients medical history and correlating clinical signs. The International Association for the Study of Pain (IASP) compiled and later on revised operational diagnostic criteria resulting in a satisfactory sensitivity and specificity for both research and clinical needs. Additionally, diagnostic tools can support the clinical suspicion - reasonable tests are conventional X-ray examination comparing sides, magnetic resonance imaging and a 3-phase bone scintigraphy. Moreover, electrophysiological examinations can prove a nerve lesion and differentiate between CRPS type I and II. A temperature difference can be detected via infrared thermography. Furthermore, quantitative sensory testing can verify the magnitude of the sensory disturbance and can be beneficial to objectify therapeutic effects. Use of these diagnostic tools, even after achievement of normal findings, cannot exclude a CRPS and the decision for therapeutic initiation should not be influenced thereby.


Evaluating Clinical Outcomes in Patients Being Treated Exclusively via Telepsychiatry: Retrospective Data Analysis.
Depression and anxiety are highly prevalent conditions in the United States. Despite the availability of suitable therapeutic options, limited access to high-quality psychiatrists represents a major barrier to treatment. Although telepsychiatry has the potential to improve access to psychiatrists, treatment efficacy in the telepsychiatry model remains unclear. Our primary objective was to determine whether there was a clinically meaningful change in 1 of 2 validated outcome measures of depression and anxiety-the Patient Health Questionnaire-8 (PHQ-8) or the Generalized Anxiety Disorder-7 (GAD-7)-after receiving at least 8 weeks of treatment in an outpatient telepsychiatry setting. We included treatment-seeking patients enrolled in a large outpatient telepsychiatry service that accepts commercial insurance. All analyzed patients completed the GAD-7 and PHQ-8 prior to their first appointment and at least once after 8 weeks of treatment. Treatments included comprehensive diagnostic evaluation, supportive psychotherapy, and medication management. In total, 1826 treatment-seeking patients were evaluated for clinically meaningful changes in GAD-7 and PHQ-8 scores during treatment. Mean treatment duration was 103 (SD 34) days. At baseline, 58.8% (1074/1826) and 60.1% (1097/1826) of patients exhibited at least moderate anxiety and depression, respectively. In response to treatment, mean change for GAD-7 was -6.71 (95% CI -7.03 to -6.40) and for PHQ-8 was -6.85 (95% CI -7.18 to -6.52). Patients with at least moderate symptoms at baseline showed a 45.7% reduction in GAD-7 scores and a 43.1% reduction in PHQ-8 scores. Effect sizes for GAD-7 and PHQ-8, as measured by Cohen d for paired samples, were d=1.30 (P<.001) and d=1.23 (P<.001), respectively. Changes in GAD-7 and PHQ-8 scores correlated with the type of insurance held by the patients. Greatest reductions in scores were observed among patients with commercial insurance (45% and 43.9% reductions in GAD-7 and PHQ-8 scores, respectively). Although patients with Medicare did exhibit statistically significant reductions in GAD-7 and PHQ-8 scores from baseline (P<.001), these improvements were attenuated compared to those in patients with commercial insurance (29.2% and 27.6% reduction in GAD-7 and PHQ-8 scores, respectively). Pairwise comparison tests revealed significant differences in treatment responses in patients with Medicare versus commercial insurance (P<.001). Responses were independent of patient geographic classification (urban vs rural; P=.48 for GAD-7 and P=.07 for PHQ-8). The finding that treatment efficacy was comparable among rural and urban patients indicated that telepsychiatry is a promising approach to overcome treatment disparities that stem from geographical constraints. In this large retrospective data analysis of treatment-seeking patients using a telepsychiatry platform, we found robust and clinically significant improvement in depression and anxiety symptoms during treatment. The results provide further evidence that telepsychiatry is highly effective and has the potential to improve access to psychiatric care.


Reducing maternal deaths in a low resource setting in Nigeria.
To assess the impact of the adoption of evidence based guidelines on maternal mortality reduction at Enugu State University Teaching Hospital, Nigeria. A retrospective review of all maternal deaths between 1 st January, 2005 and 31 st December, 2010 was carried out. Evidence based management guidelines for eclampsia and post-partum hemorrhage were adopted. These interventions strategy were carried out from 1 st January, 2008-31 st December, 2010 and the result compared with that before the interventions (2005-2007). Maternal mortality ratio (MMR) and case fatality rates. There were 9150 live births and 59 maternal deaths during the study period, giving an MMR of 645/100 000 live births. Pregnant women who had no antenatal care had almost 10 times higher MMR. There was 43.5% reduction in the MMR with the interventions (488 vs. 864/100 000 live births P = 0.039, odds ratio = 1.77). There was also significant reduction in case fatality rate for both eclampsia (15.8% vs. 2.7%; P = 0.024, odds ratio = 5.84 and Post partum hemorrhage (PPH) (13.6% vs. 2.5% P value = 0.023, odds ratio = 5.5. Obstetric hemorrhage was the most common cause of death (23.73%), followed by the eclampsia. Administration of evidence based intervention is possible in low resource settings and could contribute to a significant reduction in the maternal deaths.


Study of developed resistance due to antibiotic treatment of coagulase-negative Staphylococci.
Coagulase-negative Staphylococci (CoNS) are a major cause of postoperative infections. These infections are often associated with foreign material implants and/or a compromised immune system in the patient. Multiresistant strains are increasingly common in the hospital environment and there is concern that the infections will become difficult or impossible to treat. This report is based on a study of 75 patients, with postoperative infections caused by CoNS after thoracic surgery. All patients were treated with surgical revision and antibiotic therapy. One or more bacterial cultures were made in each case, and the resistance pattern of the CoNS found was determined. The goal of the study was to evaluate possible relationships between antibiotic therapy and the appearance of resistance to antibiotics in CoNS found. To describe this relationship, three models were constructed and analyzed by multiple logistic regression. The results indicate an increased resistance to beta-lactam antibiotics and clindamycin after the use of cephalosporins. Also, the use of vancomycin or vancomycin in combination with rifampicin or fusidic acid increases the risk for development of resistance to beta-lactam antibiotics, ciprofloxacin, fusidic acid, clindamycin, netilmycin, and rifampicin. The hypothesis that a combination of antibiotics will curtail the development of resistance was not supported in this study.


Effect of viral and bacterial pneumonias on cell-mediated immunity in humans.
Cell-mediated immunity (CMI) was assessed during infection and after convalescence in 12 patients with influenza pneumonia and 10 patients with bacterial pneumonia. The patients with influenza pneumonia had a marked impairment of skin test reactivity, and their lymphocytes showed a diminished response to phytohemagglutinin and streptokinase-streptodornase stimulation in vitro. Suppression of CMI was related to the severity of the pneumonia. Patients with bacterial pneumonia showed as great a suppression of the response to phytohemagglutinin and streptokinase-streptodornase as the patients with viral pneumonia. All parameters of CMI returned to normal in both groups after convalescence. The depression of CMI could not be related to a decrease in the number of thymus-derived lymphocytes or to serum-suppressive factors in these patients.


Why Gametes are not Like Enriched Uranium.
According to Rivka Weinberg, gametes are like enriched uranium: both are hazardous materials. Exposing human beings to enriched uranium can result in radioactivity and decreased life expectancy, while exposing sperm and ova to each other can result in the creation of needy innocent persons with full moral status. Weinberg argues that when we engage in activities that put our gametes at risk of joining with others and growing into persons, we assume the costs of that risky activity. She calls this the Hazmat Theory of parental responsibility. The theory is novel and important, and has far-reaching consequences for the ethics of procreation, parenting, and population, implying among other things that that the only way to avoid acquiring parental responsibilities may be to "abstain from sexual intercourse or surgically interfere with our gamete-release system." For these reasons the theory merits careful scrutiny. In this article I criticize the theory's account of how parental responsibility is acquired and its treatment of the standard of care expected of gamete possessors, and argue that it fails to properly account for a distinction between procreative costs and parental responsibility. Even if gametes are hazardous, it does not follow that parental responsibility in Weinberg's sense is acquired whenever one brings new persons into existence.


Ca2+ Signaling and IL-8 Secretion in Human Testicular Peritubular Cells Involve the Cation Channel TRPV2.
Peritubular cells are part of the wall of seminiferous tubules in the human testis and their contractile abilities are important for sperm transport. In addition, they have immunological roles. A proteomic analysis of isolated human testicular peritubular cells (HTPCs) revealed expression of the transient receptor potential channel subfamily V member 2 (TRPV2). This cation channel is linked to mechano-sensation and to immunological processes and inflammation in other organs. We verified expression of TRPV2 in peritubular cells in human sections by immunohistochemistry. It was also found in other testicular cells, including Sertoli cells and interstitial cells. In cultured HTPCs, application of cannabidiol (CBD), a known TRPV2 agonist, acutely induced a transient increase in intracellular Ca2+ levels. These Ca2+ transients could be blocked both by ruthenium red, an unspecific Ca2+ channel blocker, and tranilast (TRA), an antagonist of TRPV2, and were also abolished when extracellular Ca2+ was removed. Taken together this indicates functional TRPV2 channels in peritubular cells. When applied for 24 to 48 h, CBD induced expression of proinflammatory factors. In particular, mRNA and secreted protein levels of the proinflammatory chemokine interleukin-8 (IL-8/CXCL8) were elevated. Via its known roles as a major mediator of the inflammatory response and as an angiogenic factor, this chemokine may play a role in testicular physiology and pathology.


The timing and frequency of complications after peripheral percutaneous transluminal angioplasty and iliac stenting: is a change from inpatient to outpatient therapy feasible?
A prospective study was performed to assess the frequency and timing of complications after transluminal angioplasty and stent placement with a view to changing our practice and performing these procedures on an outpatient basis. A total of 266 angioplasties and 51 stent deployments were attempted on 240 consecutive patients. Immediate complications were documented by the radiologists. The timing and nature of any complications during and beyond the first 24 hr were reported by the vascular surgeons. There were 14 complications in 240 patients, giving a complication rate of 4.8% per vessel segment dilated. There were five major and nine minor complications. Eighty-six percent of complications were evident before the patient had left the angiography suite. All complications were evident within 4.5 hr of the procedure. The timing of complications suggests it would be reasonable to perform percutaneous transluminal angioplasties and iliac stenting on an outpatient basis in suitable patients.


Transcriptional regulation of a gene encoding the small subunit of ribulose-1,5-bisphosphate carboxylase in soybean tissue is linked to the phytochrome response.
The effects of white light, far-red light, and darkness on the transcription of a soybean ribulose-1,5-biphosphate carboxylase small subunit gene, SRS1, were investigated. RNA was labeled with [alpha-32P]UTP in nuclei isolated from plants grown under different conditions of light and darkness and used to probe Southern blots and dot blots. The levels of small subunit mRNA synthesis were normalized to ribosomal RNA synthesis. We demonstrate that the SRS1 gene is transcribed at a rate 16- to 32-fold higher in plants grown in the light than in those grown in darkness. Transcription of the small subunit increased dramatically when plants grown in darkness were given 30 min to 6 h of light and then leveled off after 24 to 48 h of exposure. When light-grown seedlings were exposed to greater than 2 h of darkness, a gradual decrease in transcription was detected. This decrease in transcription reached basal dark-grown levels after 48 h of exposure to darkness. The increase in transcription in etiolated seedlings treated with white light for 15 min could be reduced to basal levels if the treatment was followed by treatment with far-red light for 15 min. In addition, transcription in ligh-grown seedlings was reduced to basal levels when plants were exposed to far-red light for 15 min. The transcription of this ribulose-1,5-biphosphate carboxylase small subunit gene is strongly positively regulated by white light, is negatively regulated by far-red light, and exhibits a classic phytochrome-linked response.


Optimal feed level during the transition period to achieve faster farrowing and high colostrum yield in sows.
This study aimed to determine the optimal supply of lactation feed during the transition period to minimize farrowing duration (FD) and maximize colostrum yield (CY) and quality with the overall aim of reducing piglet mortality. A total of 48 sows were stratified for body weight and assigned to six levels of feed supply (1.8, 2.4, 3.1, 3.7, 4.3, and 5.0 kg/d) from day 108 of gestation until 24 h after the onset of farrowing. The number of total born, live-born, and stillborn piglets; birth time and birth weight of each piglet; and frequency of farrowing assistance (FA) was recorded, and blood samples were obtained from newborn piglets at birth. Live-born piglets were further weighed at 12 and 24 h after birth to record weight gain, which in turn was used to estimate intake and yield of colostrum. Colostrum samples were collected at 0, 12, 24, and 36 h after the onset of farrowing. FD was shortest (4.2 h) at intermediate (3.7 kg/d), longest (7.1 to 7.6 h) at low (1.8 and 2.4 kg/d), and intermediate (5.6 to 5.7 h) at high (4.3 and 5.0 kg/d) feed intake (P = 0.004; mean comparison). FA was lowest (0.7% to 0.8%) at intermediate feed intake (3.7 and 4.3 kg/d) and substantially elevated (4.3% to 4.7%) at both lower and higher feed intake (P = 0.01; mean comparison). The cubic contrast revealed 4.1 kg/d as the optimal feed intake to achieve the shortest FD and to minimize FA. Newborn piglets from second-parity sows were less vital than piglets from gilts as evaluated by blood biochemical variables immediately after birth. CY was greatest at 3.1 kg/d (P = 0.04), whereas the cubic contrast revealed 3.0 kg/d as the optimal feed intake to maximize CY. Concentrations of colostral components were affected by the diet, parity, and their interaction except for lactose concentrations. In conclusion, the study demonstrated the importance of proper feed level during the transition period on sow productivity. Moreover, this study estimated 4.1 and 3.0 kg/d as the optimal feed intake during the transition period to improve farrowing characteristic and CY, respectively, and these two feed intake levels supplied daily 38.8 MJ metabolizable energy (ME) and 23.9 g standardized ileal digestible (SID) lysine (3.0 kg/d) or 53.0 MJ ME and 32.7 g SID lysine (4.1 kg/d). The discrepancy of optimal feed intake for optimal farrowing and colostrum performance suggests that it may be advantageous to lower dietary lysine concentration in the diet fed prepartum.


The chloroplast view of the evolution of polyploid wheat.
Polyploid wheats comprise four species: Triticum turgidum (AABB genomes) and T. aestivum (AABBDD) in the Emmer lineage, and T. timopheevii (AAGG) and T. zhukovskyi (AAGGA(m) A(m) ) in the Timopheevi lineage. Genetic relationships between chloroplast genomes were studied to trace the evolutionary history of the species. Twenty-five chloroplast genomes were sequenced, and 1127 plant accessions were genotyped, representing 13 Triticum and Aegilops species. The A. speltoides (SS genome) diverged before the divergence of T. urartu (AA), A. tauschii (DD) and the Aegilops species of the Sitopsis section. Aegilops speltoides forms a monophyletic clade with the polyploid Emmer and Timopheevi wheats, which originated within the last 0.7 and 0.4 Myr, respectively. The geographic distribution of chloroplast haplotypes of the wild tetraploid wheats and A. speltoides illustrates the possible geographic origin of the Emmer lineage in the southern Levant and the Timopheevi lineage in northern Iraq. Aegilops speltoides is the closest relative of the diploid donor of the chloroplast (cytoplasm), as well as the B and G genomes to Timopheevi and Emmer lineages. Chloroplast haplotypes were often shared by species or subspecies within major lineages and between the lineages, indicating the contribution of introgression to the evolution and domestication of polyploid wheats.


Progress in acute myeloid leukaemia: small molecular inhibitors with small benefits.
The use of small molecule inhibitors in acute myeloid leukaemia (AML) has become ubiquitous with the US Food and Drug Administration approval of multiple agents between 2017 and 2018. Despite the promise, some of these indications are based on early efficacy data (phase I/II), and single-arm studies, and have not been tested in randomised trials. Furthermore, there are important limitations in the evidence that exists in randomised trials. This perspective aims to summarise the data that formed the basis for approval of gilteritinib, glasdegib, ivosidenib, enasidenib and venetoclax. It also aims to shed a light on some of the limitations in the evidence. Clinicians should exercise caution when using drugs that largely have yet to show an improvement in survival over the standard of care in AML.


A comparison of ovarian cancer treatments: analysis of utility assessments of ovarian cancer patients, at-risk population, general population, and physicians.
Perceptions of the severity of chemotherapy-related toxicity differ among physicians, ovarian cancer patients, at-risk individuals, and persons in the general population. In this study, we elicited assessments of toxicity from women with ovarian cancer, women at increased risk of developing ovarian cancer, women in the general population, and gynecologic oncologists. Thirty-nine ovarian cancer patients, fifteen women at increased risk, thirty-nine women at baseline risk, and eleven gynecologic oncologists completed utility assessment surveys. There was good face validity to the utility exercise as assessments of health states. Health states with toxicity were consistently associated with less favorable assessments than the health states with no toxicity. The ovarian cancer patients as a group and the women at risk for the development of ovarian cancer viewed health states with toxicity similarly and more favorably than women in the general population. However, patient assessments varied, with the most favorable assessment of life with toxicity being reported from individuals who had experienced such toxicity. Physician assessments of the impact of toxicity on overall health status were most similar to those obtained from patients who had experienced moderate to severe toxicity, and were more favorable than those elicited from patients who had not experienced any toxicity. Assessments of the impact of chemotherapy-related toxicity vary depending on the perspective of the individual responding to these questions. In discussing ovarian cancer treatments with at-risk women and the general population, concerns over treatment-related toxicity are likely to be greater than those expressed by persons who are more familiar with the actual occurrence of such events and therapeutic preferences may be affected as a result.


Gene expression profiling of papillary thyroid carcinoma identifies transcripts correlated with BRAF mutational status and lymph node metastasis.
To identify papillary thyroid carcinoma (PTC)-associated transcripts, we compared the gene expression profiles of three Serial Analysis of Gene Expression libraries generated from thyroid tumors and a normal thyroid tissue. Selected transcripts were validated in a panel of 57 thyroid tumors using quantitative PCR (qPCR). An independent set of 71 paraffin-embedded sections was used for validation using immunohistochemical analysis. To determine if PTC-associated gene expression could predict lymph node involvement, a separate cohort of 130 primary PTC (54 metastatic and 76 nonmetastatic) was investigated. The BRAF(V600E) mutational status was compared with qPCR data to identify genes that might be regulated by abnormal BRAF/MEK/extracellular signal-regulated kinase signaling. We identified and validated new PTC-associated transcripts. Three genes (CST6, CXCL14, and DHRS3) are strongly associated with PTC. Immunohistochemical analysis of CXCL14 confirmed the qPCR data and showed protein expression in PTC epithelial cells. We also observed that CST6, CXCL14, DHRS3, and SPP1 were associated with PTC lymph node metastasis, with CST6, CXCL14, and SPP1 being positively correlated with metastasis and DHRS3 being negatively correlated. Finally, we found a strong correlation between CST6 and CXCL14 expression and BRAF(V600E) mutational status, suggesting that these genes may be induced subsequently to BRAF activation and therefore may be downstream in the BRAF/MEK/extracellular signal-regulated kinase signaling pathway. CST6, CXCL14, DHRS3, and SPP1 may play a role in PTC pathogenesis and progression and are possible molecular targets for PTC therapy.


Single Administration of Melatonin Modulates the Nitroxidergic System at the Peripheral Level and Reduces Thermal Nociceptive Hypersensitivity in Neuropathic Rats.
Neuropathic pain is a severe condition with unsatisfactory treatments. Melatonin, an indolamine, seems to be a promising molecule suitable for this purpose due to its well-known anti-inflammatory, analgesic, and antioxidant effects, as well as its modulation of the nitroxidergic system. Nevertheless, the data on its mechanism of action and potentialities are currently insufficient in this pathology, especially at the peripheral level. Thus, this work evaluated the effect of a single administration of melatonin in an established mononeuropathy pain model that monitors the behaviour and the changes in the nitroxidergic system in dorsal root ganglia and skin, which are affected by nervous impairment. Experiments were carried out on Sprague Dawley rats subdivided into the sham operated (control) and the chronic constriction injured animals, a model of peripheral neuropathic pain on sciatic nerve. Single administrations of melatonin (5-10 mg/kg) or vehicle were injected intraperitoneally on the 14th day after surgery, when the mononeuropathy was established. The animals were behaviourally tested for thermal hyperalgesia. The dorsal root ganglia and the plantar skin of the hind-paws were removed and processed for the immunohistochemical detection of neuronal and inducible nitric oxide synthases. The behavioural results showed an increase of withdrawal latency during the plantar test as early as 30 min after melatonin administration. The immunohistochemical results indicated a modulation of the nitroxidergic system both at dorsal root ganglia and skin level, permitting speculate on a possible mechanism of action. We showed that melatonin may be a possible therapeutic strategy in neuropathic pain.


[Chemical characterisation and serological activity of lipopolysaccharides of Ralstonia solanacearum].
Lipopolysaccharides (LPS) of 8 strains of Ralstonia solanacearum (767, 4157, 5712, 7944, 7945, 7955, 8089 and 8110) characterized by different types of the structures of O-specific polysaccharide chains have been investigated. The ELISA and ELISA-inhibition methods were used to investigate the extent of antigenic activity of the presented LPS with respect to the obtained polyclonal O-antisera to the above strains. When analyzing the results obtained the strains were divided into five serogroups as the result of biochemical and serological investigations. The first serogroup included the strains 5712, 7945, 7955 and 8110, whose LPS and O-antisera were characterized by the high level of serological affinity. The second and the third serogroups included the strains 767 and 7944, respectively. Their LPS demonstrated antigenic specificity in the homologous systems only, as well as to O-serum to the strain 5712. The fourth and the fifth serogroups included strains 4157 and 8089, respectively, whose LPS did not manifest the antigenic specificity with respect to O-antisera of the other strains, besides their own ones, and which were characterized by different in principle types of the O-PS structures O-PS.


3D Printed Graphene and Graphene/Polymer Composites for Multifunctional Applications.
Three-dimensional (3D) printing, alternatively known as additive manufacturing, is a transformative technology enabling precise, customized, and efficient manufacturing of components with complex structures. It revolutionizes traditional processes, allowing rapid prototyping, cost-effective production, and intricate designs. The 3D printed graphene-based materials combine graphene's exceptional properties with additive manufacturing's versatility, offering precise control over intricate structures with enhanced functionalities. To gain comprehensive insights into the development of 3D printed graphene and graphene/polymer composites, this review delves into their intricate fabrication methods, unique structural attributes, and multifaceted applications across various domains. Recent advances in printable materials, apparatus characteristics, and printed structures of typical 3D printing techniques for graphene and graphene/polymer composites are addressed, including extrusion methods (direct ink writing and fused deposition modeling), photopolymerization strategies (stereolithography and digital light processing) and powder-based techniques. Multifunctional applications in energy storage, physical sensor, stretchable conductor, electromagnetic interference shielding and wave absorption, as well as bio-applications are highlighted. Despite significant advancements in 3D printed graphene and its polymer composites, innovative studies are still necessary to fully unlock their inherent capabilities.


Detection of gonococcal antigens by an indirect sandwich enzyme-linked immunosorbent assay.
The detection of gonococcal antigens by an indirect sandwich ELISA system is described. The feasibility of using rabbit antiserum raised against whole cells of Neisseria gonorrhoeae strain 9 to detect gonococcal lipopolysaccharide, whole cells, and outer membrane (OM) protein was investigated. OM protein was found to be the main antigen detectable with the antiserum in a direct ELISA system. A positive result in the indirect assay could be obtained with a minimum of 46 to 92 ng of gonococcal OM protein or with 6.6 X 10(3) cfu of N. gonorrhoeae. The sensitivity of the assay was found to be approximately eightfold lower with OM complex from a strain of N. meningitidis serogroup B for which the minimum amount of OM protein detected was 375 ng. Negative results were obtained with OM complex from Streptococcus agalactiae, Bacteroides bivius and Escherichia coli. The assay seems to be highly specific for gonococcal antigens. The sensitivity of the assay and its specificity commend it for further evaluation in the detection of gonococcal antigens in clinical specimens.


Inhibition of mammary duct development but not alveolar outgrowth during pregnancy in transgenic mice expressing active TGF-beta 1.
The transforming growth factors beta (TGFs-beta) are potent inhibitors of cell proliferation and are usually secreted in a latent form. TGF-beta 1, TGF-beta 2, and TGF-beta 3 are expressed in distinct but overlapping patterns in the developing mouse mammary gland. To study the role of transforming growth factor-beta 1 (TGF-beta 1) in normal mammary development and in mammary neoplasia, we have constructed three transgenic mouse lines that express a simian TGF-beta 1 s223/225 mutated to produce a constitutively active product under the control of the MMTV enhancer/promoter. Expression of the transgene, as confirmed by in situ hybridization, immunohistochemistry, and Northern blot analysis, was associated with marked suppression of the normal pattern of mammary ductal tree development in female transgenics. Reduction in total ductal tree volume was observed at 7 weeks, soon after estrous begins, and was most apparent at 13 weeks, as ductal growth in the normal mammary gland declines. This effect was seen in all three lines. However, during pregnancy, alveolar outgrowths developed from the hypoplastic ductal tree, and lactation occurred, therefore, all transgenic females could feed full litters. Unlike many other transgenic mouse models in which expression of growth factors or oncogenes under control of the MMTV promoter leads to mammary epithelial hyperplasia and increased tumor formation, the MMTV-TGF-beta 1S223/225 transgene causes conditional hypoplasia of the mammary ductal tree and no spontaneous tumors have been detected in the MMTV-TGF-beta 1S223/225 transgenic animals.


Ligand and oxidation-state specific regulation of the heme-based oxygen sensor FixL from Sinorhizobium meliloti.
Phosphorylation of the transcription factor RmFixJ is the key step in the hypoxic induction of Sinorhizobium meliloti nitrogen fixation genes. Oxygen regulates this process by binding reversibly to RmFixL, a heme protein kinase whose deoxy form catalyzes the phosphoryl transfer from ATP to RmFixJ. Here we present the first quantitative measure of the extent by which various heme ligands inhibit the turnover of RmFixJ to phospho-RmFixJ. We also quantitate the inhibition by ligands of the reaction of RmFixL with ATP, in the absence of RmFixJ, to form phospho-RmFixL, i.e., the "autophosphorylation". Phospho-RmFixL formed from autophosphorylation will transfer its phosphoryl group to RmFixJ in an oxygen-independent "phosphotransfer." Here we show that the mode of substrate presentation, i.e., simultaneous versus sequential, influences the regulation of phosphoryl transfer by heme status. Inhibition factors for O(2), CO, NO, CN(-), and imidazole in the presence of RmFixJ are drastically different from the inhibition of autophosphorylation by the same ligands. Oxidation of the heme iron in unliganded RmFixL is known to have no effect on either of the sequential reactions; yet oxidation causes a 100-fold decrease in RmFixJ turnover when ATP and RmFixJ are presented simultaneously. The profound difference between the regulation of isolated RmFixL versus the complex of RmFixL with RmFixJ shows that interaction of a response regulator with its histidine-kinase partner need not be limited to the enzymatic regions of the histidine kinase, but can extend also to its sensory domain.


Compensatory Hyperhidrosis After Non-Surgical Treatment of Primary Focal Hyperhidrosis: Two-Year Single-Centered Prospective Study From Jordan.
Primary focal hyperhidrosis (PH) can be managed by a wide range of medical and surgical modalities. Compensatory hyperhidrosis (CH) is a well-documented complication of surgical treatment. We aimed to investigate the occurrence of compensatory hyperhidrosis (CH) in PH patients after nonsurgical treatment with botulinum toxin A (BTX- A) or iontophoresis. We carried out a unicentric prospective study on PH patients from King Abdullah University Hospital (KAUH) in Jordan. PH patients were evaluated after 1-month of nonsurgical treatment. Patients who developed CH were re-assessed after 3-6 months through a telephone-based interview. A total of 86 patients with PH who underwent nonsurgical treatment with iontophoresis or botulinum toxin were recruited. Twenty-four (27.9%) patients developed subjective CH. It was mild in (75%), moderate in (21%), and severe in (4%) of patients affected, it was self-limiting within a few months in all patients. Patients with CH did not differ significantly in demographic or clinical variables from patients who did not develop CH except at the site of PH (p value = .05). The findings of this study indicate that more than quarter (27.9%) of patients with PH may develop minor compensatory sweating, however this didn't affect satisfaction with treatment.


Effects of enhanced bioturbation intensities on the toxicity assessment of legacy-contaminated sediments.
Many benthic communities within estuarine ecosystems are highly degraded due to the close proximity of urban and industrial contamination sources. The maintenance of recolonised, healthy ecosystems following remediation is a challenge, and better techniques are required for monitoring their progressive recovery. Rates of ecosystem recovery are influenced by the changes in the concentrations and forms of contaminants, the sensitivity of recolonising organisms to bioavailable contaminants, and a range of abiotic and biotic factors influencing the exposure of organisms to the contamination. Here we investigate the influence of bioturbation by an active amphipod (Victoriopisa australiensis) on the bioavailability of metals and hydrocarbons in highly contaminated sediments. Changes in contaminant bioavailability were evaluated by assessing sublethal effects to a smaller cohabiting amphipod (Melita plumulosa). For predominantly metal-contaminated sediments, the presence of V. australiensis generally increased survival and reproduction of M. plumulosa when compared to treatments with only M. plumulosa present (from 42 to 93% survival and 3-61% reproduction). The decrease in toxic effects to M. plumulosa corresponded with lower dissolved copper and zinc concentrations in the overlying waters (14-9 μg Cu L-1, and 14 to 6 μg Zn L-1 for absence to presence of V. australiensis). For sediments contaminated with both hydrocarbons and metals, the increased bioturbation intensity by V. australiensis resulted in decreased reproduction of M. plumulosa, despite lower dissolved metal exposure, and indicated increased bioavailability of the hydrocarbon contaminants. Thus, the presence of a secondary active bioturbator can enhance or suppress toxicity to co-inhabiting organisms, and may depend on the contaminant class and form. The results highlight the need to consider both abiotic and biotic interactions when using laboratory studies to evaluate the ability of organisms to recolonise and reproduce within benthic environments degraded by contamination, or for more general extrapolation for sediment quality assessment purposes.


Aberrantly high glycated haemoglobin measurement due to the haemoglobin variant Hb Santa Juana.
Various laboratory and patient-related factors can result in falsely high or low glycated haemoglobin (HbA1c) measurements, and haemoglobin (Hb) variants that interfere with laboratory readings is an important cause of this. We report a case of a rare Hb variant, Hb Santa Juana, manifesting as a falsely high HbA1c in a 62-year-old patient with type 2 diabetes mellitus. The patient presented with high HbA1c values that persisted despite the intensification of anti-diabetic treatment. His home blood glucose levels were incongruently low compared to his HbA1c values. Further investigations revealed a family history of the variant Hb Santa Juana. This was confirmed in the patient when his blood was sent for DNA analysis. It is important for clinicians to be aware of the factors that can influence laboratory HbA1c measurements, as clinical decisions on treatment are often based on these measurements.


Adaptive control deficits in attention-deficit/hyperactivity disorder (ADHD): the role of error processing.
Cognitive performance of children with attention-deficit hyperactivity disorder (ADHD) is characterized by large moment-to-moment fluctuations in cognitive control reflected by a highly inconsistent and inaccurate response style. It has been suggested that abnormal error processing underlies this failure to implement adequate control. We investigated the error-related negativity (ERN), a negative deflection in the event-related potential (ERP) time-locked to erroneous responses in 16 rigorously screened ADHD boys aged 8-12 years and 16 age-matched normal control boys during a modified Eriksen flanker paradigm with two levels of time pressure. Children with ADHD responded as fast and regularly as controls, but committed significantly more errors, particularly when facing time pressure and response conflict. ADHD children produced shorter runs of correct responses than controls. In addition, with high time pressure, error runs were prolonged relative to control children, suggesting an increase in both frequency and magnitude of temporary lapses of control. ERP amplitude differences between correct and incorrect responses were diminished in ADHD children, whereas post-error slowing remained unaffected. This pattern of results indicates that a specific deficit in monitoring ongoing behaviour, rather than insufficient strategic adjustments, gave rise to performance limitations in ADHD. Findings are discussed in terms of anterior cingulate cortex (ACC) dysfunction, leading to a failure to predict the likelihood that an error occurs in a given context.


Predicting short-term disease progression among HIV-infected patients in Asia and the Pacific region: preliminary results from the TREAT Asia HIV Observational Database (TAHOD).
HIV disease progression has been well documented in Western populations. This study aimed to estimate the short-term risk of AIDS and death from the TREAT Asia HIV Observational Database (TAHOD), a prospective, multicentre cohort study in Asia and the Pacific region. Prospective data were analysed to estimate short-term disease progression. Endpoints were defined as the time from study entry to diagnosis with AIDS or death. Antiretroviral treatment was fitted as a time-dependent variable. Predictors of disease progression were assessed using Cox proportional hazards models, and prognostic models were developed using Weibull models. A total of 1260 patients with prospective follow-up data contributed 477 person-years of follow-up, during which 18 patients died and 34 were diagnosed with AIDS, a combined rate of 10.1 per 100 person-years. Compared with patients receiving antiretroviral treatment, patients not on treatment had a higher rate of disease progression (17.6 vs. 8.1 per 100 person-years, respectively). Baseline CD4 count was the strongest predictor of disease progression. Prognostic models, using either a baseline CD4 count as the sole marker or markers including baseline haemoglobin, AIDS-related symptoms and previous or current antiretroviral treatment, were successful at identifying patients at high risk of short-term disease progression. Similar to the situation in Western countries, baseline CD4 count was the strongest predictor of short-term disease progression. Prognostic models based on readily available clinical data and haemoglobin level should be useful in estimating short-term clinical risk in HIV-infected patients in Asia and the Pacific region.


A laser Doppler system for monitoring of intracerebral microcirculation.
A two-channel standard laser Doppler perfusion monitor has been adapted for intracerebral measurements. Software developed in Labview makes it possible to present the microvascular perfusion, total light intensity (TLI), heart rate and trend curves in real-time during surgery. A custom-made optical probe was designed in order to enable easy fixation during brain surgery. The constructed brain probe was evaluated and compared to a standard probe. Both probes presented similar feasibility when used for the skin recordings. In addition, evaluation was done in one patient in relation to tumor resection. Stable perfusion and TLI signals were immediately recorded when the probe was positioned in cerebral tissue. Movement artifacts were clearly seen when the probe was moved to a new site. Recordings in cortex and tumor border showed higher perfusion and lower TLI compared to measurements in subcortical white matter. The calculated heart rate estimate agreed well with the noted value from the electrocardiographic patient monitoring system.


Index suffix-prefix overlaps by (w, k)-minimizer to generate long contigs for reads compression.
Advanced high-throughput sequencing technologies have produced massive amount of reads data, and algorithms have been specially designed to contract the size of these datasets for efficient storage and transmission. Reordering reads with regard to their positions in de novo assembled contigs or in explicit reference sequences has been proven to be one of the most effective reads compression approach. As there is usually no good prior knowledge about the reference sequence, current focus is on the novel construction of de novo assembled contigs. We introduce a new de novo compression algorithm named minicom. This algorithm uses large k-minimizers to index the reads and subgroup those that have the same minimizer. Within each subgroup, a contig is constructed. Then some pairs of the contigs derived from the subgroups are merged into longer contigs according to a (w, k)-minimizer-indexed suffix-prefix overlap similarity between two contigs. This merging process is repeated after the longer contigs are formed until no pair of contigs can be merged. We compare the performance of minicom with two reference-based methods and four de novo methods on 18 datasets (13 RNA-seq datasets and 5 whole genome sequencing datasets). In the compression of single-end reads, minicom obtained the smallest file size for 22 of 34 cases with significant improvement. In the compression of paired-end reads, minicom achieved 20-80% compression gain over the best state-of-the-art algorithm. Our method also achieved a 10% size reduction of compressed files in comparison with the best algorithm under the reads-order preserving mode. These excellent performances are mainly attributed to the exploit of the redundancy of the repetitive substrings in the long contigs. https://github.com/yuansliu/minicom. Supplementary data are available at Bioinformatics online.


Bcl2 retards G1/S cell cycle transition by regulating intracellular ROS.
Bcl2's antiapoptotic function is regulated by phosphorylation. Bcl2 also regulates cell cycle progression, but the molecular mechanism is unclear. Bcl2 is functionally expressed in mitochondria where it can act as an antioxidant that may regulate intracellular reactive oxygen species (ROS). Since ROS have been reported to act as second messengers in cell signaling, we tested whether Bcl2 phosphorylation regulates ROS and cell cycle progression. G1 --> S transition and ROS levels were measured in cells expressing either the gain of function phosphomimetic Bcl2 mutants S70E and T69E/S70E/S87E (EEE) or the nonphosphorylatable and survival-deficient mutants S70A and T69A/S70A/S87A (AAA). Expression of S70E and EEE but not the A-containing Bcl2 mutants retards G1 --> S transition by 35% to 50% and significantly slows cell growth in association with reduced levels of intracellular ROS. In addition to expression of the phosphomimetic Bcl2 mutants, either interleukin-3 withdrawal or treatment of cells with the antioxidant pyrrolidine dithiocarbamate (PDTC) also reduces intracellular ROS levels in association with up-regulation of p27 and accumulation of cells in G0/G1. Retardation of G1 --> S transition can be overridden by directly adding H2O2 to the cells in a mechanism that involves down-regulation of p27 and activation of Cdk2. Thus, Bcl2 may regulate G1 --> S transition by a novel signaling mechanism that couples regulation of intracellular ROS with p27 and Cdk2. Furthermore, phosphorylation of Bcl2 may functionally link its antiapoptotic, cell cycle retardation, and antioxidant properties.


Density-dependent selection incorporating intraspecific competition. II. A diploid model.
A diploid model is introduced and analyzed in which intraspecific competition is incorporated within the context of density-regulated selection. It is assumed that each genotype has a unique carrying capacity corresponding to the equilibrium population size when only that type is present. Each genotypic fitness at a single diallelic autosomal locus is a decreasing function of a distinctive effective population size perceived as a result of intraspecific competition. The resulting fitnesses are both density and frequency dependent with selective advantage determined by a balance between genotypic carrying capacity and sensitivity to intraspecific competition. A major finding is that intergenotypic interactions may allow genetic variation to be more easily maintained than in the corresponding model of purely density-dependent selection. In addition, numerical study confirms the possible existence of multiple interior equilibria and that neither overdominance in fitness nor carrying capacity is necessary for stability. The magnitude of the equilibrium population size and optimization principles are also discussed.


The Latitudes, Attitudes, and Platitudes of Watershed Phosphorus Management in North America.
Phosphorus (P) plays a crucial role in agriculture as a primary fertilizer nutrient-and as a cause of the eutrophication of surface waters. Despite decades of efforts to keep P on agricultural fields and reduce losses to waterways, frequent algal blooms persist, triggering not only ecological disruption but also economic, social, and political consequences. We investigate historical and persistent factors affecting agricultural P mitigation in a transect of major watersheds across North America: Lake Winnipeg, Lake Erie, the Chesapeake Bay, and Lake Okeechobee/Everglades. These water bodies span 26 degrees of latitude, from the cold climate of central Canada to the subtropics of the southeastern United States. These water bodies and their associated watersheds have tracked trajectories of P mitigation that manifest remarkable similarities, and all have faced challenges in the application of science to agricultural management that continue to this day. An evolution of knowledge and experience in watershed P mitigation calls into question uniform solutions as well as efforts to transfer strategies from other arenas. As a result, there is a need to admit to shortcomings of past approaches, plotting a future for watershed P mitigation that accepts the sometimes two-sided nature of Hennig Brandt's "Devil's Element."


[A concept of need and construction of a hip joint prosthesis with isolation of the friction point from the tissue].
The paper presents data available in the literature and the author's own data on the causes of bone tissue resolution around endoprostheses and on the occurrence of their aseptic instability. The author's own clinical data on 546 operations and follow-ups up to 16 years are strong evidence and his experimental findings are unique in the number of animals operated on and follow-up periods. The material was examined roentgenologically, morphologically, spectroscopically and the mineral content of bone tissue microstructures were determined. With the methods applied, the author clearly shows the role of tissue metallosis in the occurrence of aseptic instability of prosthesis. The author presents two up-to-day constructions of the total-type endoprostheses with the friction point isolated from tissue. The prostheses are accessible for manufacture at the modern enterprises and require no materials in short supply.


Tele-Robotic Platform for Dexterous Optical Single-Cell Manipulation.
Single-cell manipulation is considered a key technology in biomedical research. However, the lack of intuitive and effective systems makes this technology less accessible. We propose a new tele-robotic solution for dexterous cell manipulation through optical tweezers. A slave-device consists of a combination of robot-assisted stages and a high-speed multi-trap technique. It allows for the manipulation of more than 15 optical traps in a large workspace with nanometric resolution. A master-device (6+1 degree of freedom (DoF)) is employed to control the 3D position of optical traps in different arrangements for specific purposes. Precision and efficiency studies are carried out with trajectory control tasks. Three state-of-the-art experiments were performed to verify the efficiency of the proposed platform. First, the reliable 3D rotation of a cell is demonstrated. Secondly, a six-DoF teleoperated optical-robot is used to transport a cluster of cells. Finally, a single-cell is dexterously manipulated through an optical-robot with a fork end-effector. Results illustrate the capability to perform complex tasks in efficient and intuitive ways, opening possibilities for new biomedical applications.


Developmentally regulated expression of the murine ortholog of the potassium channel KIR4.2 (KCNJ15).
The gene KIR4.2 (K(+) inwardly rectifying channel 4.2) has been recently identified in the Down syndrome Chromosome Region 1. We have cloned the mouse ortholog of KIR4.2 and characterized its expression pattern. In situ hybridization showed a restricted and developmentally regulated pattern of expression. The expression is starting at E12.5 and expands at E14.5 in different tissues and organs, which may be affected in Down syndrome: heart, thymus, thyroid gland, and perichondrium. At E17.5, additional epithelia (kidney, bladder, stomach, lung) expressed also strongly the gene.


Development of highly efficient protocols for extraction and amplification of cytomegalovirus DNA from dried blood spots for detection and genotyping of polymorphic immunomodulatory genes.
Congenital cytomegalovirus (CMV) infection is a major cause of birth defects ranging from developmental disorders to stillbirth. Most newborns affected by CMV do not present with symptoms at birth but are at risk of sequelae at later stages of their childhood. Stored dried blood spots (DBS) taken at birth can be used for retrospective diagnosis of hereditary diseases, but detection of pathogens is challenged by potentially low pathogen concentrations in the small blood volume available in a DBS. Here we test four different extraction methods for optimal recovery of CMV DNA from DBS at low to high CMV titers. The recovery efficiencies varied widely between the different extractions (from 3% to 100%) with the most efficient method extracting up to 113-fold more CMV DNA than the least efficient and 8-fold more than the reference protocol. Furthermore, we amplified four immunomodulatory CMV genes from the extracted DNA: the UL40 and UL111A genes which occur as functional knockouts in some circulating CMV strains, and the highly variable UL146 and US28 genes. The PCRs specifically amplified the CMV genes at all tested titers with sufficient quality for sequencing and genotyping. In summary, we here report an extraction method for optimal recovery of CMV DNA from DBSs that can be used for both detection of CMV and for genotyping of polymorphic CMV genes in congenital CMV infection.


An empirically based model for latitudinal gradient in vole population dynamics.
Vole dynamics in northern Europe exhibit a well-defined geographical gradient, with oscillatory populations being confined to high latitudes. It has been proposed that oscillations in northern vole populations are driven by their interaction with specialist predators (weasels), while the more southern rodent populations are relatively stable because of regulation by generalist predators. We tested this generalist/specialist predation hypothesis by constructing an empirically based model for vole population dynamics, estimating its parameters, and making predictions about the quantitative pattern of the latitudinal shift in vole dynamics. Our results indicated that the model accurately predicted the latitudinal shift in the amplitude and periodicity of population fluctuations. Moreover, the model predicted that vole dynamics should shift from stable to chaotic as latitude is increased, a result in agreement with nonlinear time-series analysis of the data. The striking success of the model at predicting the shifts in amplitude and stability along the geographical gradient in northern Europe provides strong support for the key role of specialist and generalist predators in vole population dynamics.


[A method of tubular resection of the stomach with formation of a rectangular oblique gastro-jejunal anastomosis].
The method of tubular resection of the stomach with deomentization, subtotal resection of the lesser curvature and formation of the retrocolic right-angled gastro-jejunal anastomosis on a short loop with an oblique section of the jejunum was developed experimentally and successfully used in 98 patients for the treatment of complicated forms of gastroduodenal ulcers and of oncological patients with the tumorous involvement of the outlet portion of the stomach.


Both Male and Female Malfunction Contributes to Yield Reduction under Water Stress during Meiosis in Bread Wheat.
Water stress during meiosis in wheat is a major constraint to yield especially for the rainfed farming regions. Pollen sterility has been proposed as the most sensitive process leading to low seed set (low % of fertile spikelets), but here we show this is not universal, and that the development of female reproductive parts is equally if not more sensitive than male parts in many wheat cultivars. The first experiment examined water stress during meiosis in 46 wheat genotypes. The reduction in seed set varied widely, ranging from 6 to 48%. The second experiment differentiated the effect of water stress on the male or the female reproductive part in 13 wheat genotypes. Water stress was imposed during meiosis, with plants cross-pollinated 5 days later with pollen from stressed or unstressed plants used to pollinate emasculated stressed or unstressed female parts. Seed set and kernel weight were measured at maturity. Contrary to the well-held view that the male reproductive part is the major contributor to seed set reduction when water stress is experienced during meiosis, the stressed-female part was also a predominant contributor in four wheat genotypes among the 13 genotypes examined. This strongly indicates that both male and female parts are responsible for yield reduction when water-stressed during meiosis and suggests that it may be possible to breed tolerant wheat cultivars combining tolerance from both male and female reproductive parts.


Ertugliflozin: a sodium-glucose cotransporter-2 (SGLT-2) inhibitor for glycemic control in type 2 diabetes.
Recently, a new class of anti-diabetic drugs has found its way into the market: sodium-glucose cotransporters (SGLT) inhibitors. Two major SGLT isoforms have been identified: SGLT-2, mainly expressed in proximal renal tubules, and SGLT-1, mainly expressed in the small intestine, the proximal renal tubule, and the myocardium. SGLT-2 inhibitors increase urinary glucose excretion, lowering glycemia without inducing excessive insulin secretion. Marketed SGLT-2 inhibitors actually include dapagliflozin, canagliflozin, and empagliflozin; a new SGLT-2 inhibitor is being studied: ertugliflozin. Ertugliflozin is a potent inhibitor of SGLT-2 and possesses a high selectivity over glucose transport via SGLT-1 and several other glucose transporters GLUT-1-4. Ertugliflozin inhibits renal glucose reabsorption resulting in urinary glucose excretion and thereby reduces plasma glucose and glycated hemoglobin in subjects with type 2 diabetes mellitus. Ertugliflozin is being developed as an adjunct to diet and exercise to improve glycemic control in patients with type 2 diabetes mellitus. The aim of this review is to evaluate the preliminary published data about this new molecule.


Renal artery repair: consequence of operative failures.
This report examines the blood pressure and renal function response in 20 consecutive patients after secondary renal revascularization following failed operative repair. Most reports describing operative failure of renal artery (RA) repair emphasize the technical aspects of redo RA reconstruction and the immediate blood-pressure response to secondary operation. This report examines the eventual renal function and estimated survival after secondary intervention. Primary methods of RA reconstruction, primary blood pressure and renal function responses, and causes of failed RA repair were defined for 20 patients requiring reoperation for recurrent hypertension or renal insufficiency. These parameters were compared with secondary procedures and eventual blood pressure and renal function response. The eventual outcome for these 20 patients was compared with 514 patients managed by primary renal revascularization during the same period. Failure of primary RA repair correlated with complex fibromuscular dysplasia requiring branch ex vivo reconstruction (p = 0.020). RA thrombosis frequently required nephrectomy (83%), whereas RA stenosis was successfully reconstructed (91 %; p = 0.001). Primary and secondary blood-pressure responses were equivalent (94% vs. 95% cured or improved); however, primary and eventual renal function responses differed significantly (p = 0.015), with seven patients dialysis-dependent on follow-up. Eventual dialysis dependence was associated with preoperative azotemia (p = 0.022), bilateral failure of primary RA repair (p = 0.007), and an increased risk of follow-up death (p = 0.002). Considering all 534 patients, failed RA repair demonstrated a significant and independent association with eventual dialysis dependence and decreased dialysis-free survival. Contemporary rates of reoperation after surgical RA repair are low. In properly selected patients, beneficial blood-pressure response is reliably observed after both primary and secondary operative procedures. However, secondary procedures are associated with a significant and independent risk of eventual dialysis dependence.


Effects of Diesel Exhaust Particles on Mouse Gastric Stem Cells.
Stem cells have attracted many scientists because of their unique properties and therapeutic applications. However, very little is known on the environmental toxins that could affect their biological features. This study focuses on the consequences of the exposure of a cell line representative of the mouse gastric stem/progenitor (mGS) cells to diesel exhaust particles (DEPs). These immortal cells were cultured using routine protocols. The DEPs were added to the culture media at 1, 10, and 100 µg/mL for 1 to 72 h. The cells were assayed for their viability, migration, oxidative stress, and the expression of genes specific for cell proliferation, pluripotency, and death. DEPs induced a reduction in the metabolic activity of mGS cells, only at a high concentration of 100 µg/mL. However, no significant effects were detected on cell migration, oxidative stress markers (glutathione and thiobarbituric acid reactive substances), and cell death related proteins/genes. Interestingly, these findings were associated with down-regulation of Notch 2 and 3 and Bmi-1 proteins and activation of STAT3 involved in the regulation of the fate of stem cells. In conclusion, this study demonstrates that mGS cells have some resistance to oxidative stress and apoptosis when exposed to DEPs at the expense of their stemness.


Beyond potentiometry: robust electrochemical ion sensor concepts in view of remote chemical sensing.
For about 100 years, potentiometry with ion-selective electrodes has been one of the dominating electroanalytical techniques. While great advances in terms of selective chemistries and materials have been achieved in recent years, the basic manner in which ion-selective membranes are used has not fundamentally changed. The potential readings are directly co-dependent on the potential at the reference electrode, which requires maintenance and for which very few accepted alternatives have been proposed. Fouling or clogging of the exposed electrode surfaces will lead to changes in the observed potential. At the same time, the Nernst equation predicts quite small potential changes, on the order of millivolts for concentration changes on the order of a factor two, making frequent recalibration, accurate temperature control and electrode maintenance key requirements of routine analytical measurements. While the relatively advanced selective materials developed for ion-selective sensors would be highly attractive for low power remote sensing application, one should consider solutions beyond classical potentiometry to make this technology practically feasible. This paper evaluates some recent examples that may be attractive solutions to the stated problems that face potentiometric measurements. These include high-amplitude sensing approaches, with sensitivities that are an order of magnitude larger than predicted by the Nernst equation; backside calibration potentiometry, where knowledge of the magnitude of the potential is irrelevant and the system is evaluated from the backside of the membrane; controlled current coulometry with ion-selective membranes, an attractive technique for calibration-free reagent delivery without the need for standards or volumetry; localized electrochemical titrations at ion-selective membranes, making it possible to design sensors that directly monitor parameters such as total acidity for which volumetric techniques were traditionally used; and controlled potential coulometry, where all ions of interest are selectively transferred into the ion-selective organic phase, forming a calibration-free technique that would be exquisitely suitable for remote sensing applications.


Centriole movements in mammalian epithelial cells during cytokinesis.
In cytokinesis, when the cleavage furrow has been formed, the two centrioles in each daughter cell separate. It has been suggested that the centrioles facilitate and regulate cytokinesis to some extent. It has been postulated that termination of cytokinesis (abscission) depends on the migration of a centriole to the intercellular bridge and then back to the cell center. To investigate the involvement of centrioles in cytokinesis, we monitored the movements of centrioles in three mammalian epithelial cell lines, HeLa, MCF 10A, and the p53-deficient mouse mammary tumor cell line KP-7.7, by time-lapse imaging. Centrin1-EGFP and alpha-Tubulin-mCherry were co-expressed in the cells to visualize respectively the centrioles and microtubules. Here we report that separated centrioles that migrate from the cell pole are very mobile during cytokinesis and their movements can be characterized as 1) along the nuclear envelope, 2) irregular, and 3) along microtubules forming the spindle axis. Centriole movement towards the intercellular bridge was only seen occasionally and was highly cell-line dependent. These findings show that centrioles are highly mobile during cytokinesis and suggest that the repositioning of a centriole to the intercellular bridge is not essential for controlling abscission. We suggest that centriole movements are microtubule dependent and that abscission is more dependent on other mechanisms than positioning of centrioles.


Requirements for accurate and efficient mRNA 3' end cleavage and polyadenylation of a simian virus 40 early pre-RNA in vitro.
Using a pre-RNA containing the simian virus 40 early introns and poly(A) addition site, we investigated several possible requirements for accurate and efficient mRNA 3' end cleavage and polyadenylation in a HeLa cell nuclear extract. Splicing and 3' end formation occurred under the same conditions but did not appear to be coupled in any way in vitro. Like splicing, 3' end cleavage and polyadenylation each required Mg2+, although spermidine could substitute in the cleavage reaction. Additionally, cleavage of this pre-RNA, but not others, was totally blocked by EDTA, indicating that structural features of pre-RNA may affect the ionic requirements of 3' end formation. The ATP analog 3' dATP inhibited both cleavage and polyadenylation even in the presence of ATP, possibly reflecting the coupled nature of these activities. A 5' cap structure appears not to be required for mRNA 3' end processing in vitro because neither the presence or absence of a 5' cap on the pre-RNA nor the addition of cap analogs to reaction mixtures had any effect on the efficiency of 3' end processing. Micrococcal nuclease pretreatment of the nuclear extract inhibited cleavage and polyadenylation. However, restoration of activity was achieved by addition of purified Escherichia coli RNA, suggesting that the inhibition caused by such a nuclease treatment was due to a general requirement for mass of RNA rather than to destruction of a particular nucleic acid-containing component such as a small nuclear ribonucleoprotein.


Plasma Nervonic Acid Is a Potential Biomarker for Major Depressive Disorder: A Pilot Study.
Diagnostic biomarkers of major depressive disorder, bipolar disorder, and schizophrenia are urgently needed, because none are currently available. We performed a comprehensive metabolome analysis of plasma samples from drug-free patients with major depressive disorder (n=9), bipolar disorder (n=6), schizophrenia (n=17), and matched healthy controls (n=19) (cohort 1) using liquid chromatography time-of-flight mass spectrometry. A significant effect of diagnosis was found for 2 metabolites: nervonic acid and cortisone, with nervonic acid being the most significantly altered. The reproducibility of the results and effects of psychotropic medication on nervonic acid were verified in cohort 2, an independent sample set of medicated patients [major depressive disorder (n=45), bipolar disorder (n=71), schizophrenia (n=115)], and controls (n=90) using gas chromatography time-of-flight mass spectrometry. The increased levels of nervonic acid in patients with major depressive disorder compared with controls and patients with bipolar disorder in cohort 1 were replicated in the independent sample set (cohort 2). In cohort 2, plasma nervonic acid levels were also increased in the patients with major depressive disorder compared with the patients with schizophrenia. In cohort 2, nervonic acid levels were increased in the depressive state in patients with major depressive disorder compared with the levels in the remission state in patients with major depressive disorder and the depressive state in patients with bipolar disorder. These results suggested that plasma nervonic acid is a good candidate biomarker for the depressive state of major depressive disorder.


Tissue transglutaminase antibodies in celiac disease, comparison of an enzyme linked immunosorbent assay and a dot blot assay.
The purpose of our study is to determine the sensitivity, specificity and predictive values of an enzyme linked immunosorbent assay (ELISA) and a dot blot assay for the detection of IgA class anti-tissue transglutaminase antibodies (IgA-AtTGA) and to compare these results with those of IgA class anti-endomysium antibodies (IgA-AEA), IgA class anti-reticulin antibodies (IgA-ARA) and IgA class anti-gliadin antibodies (IgA-AGA). Serum samples from 143 patients (97 children, 46 adults) with untreated celiac disease (CD) confirmed by intestinal biopsy and 74 disease controls (64 children, 10 adults) were studied. Methods. - The anti-tissue transglutaminase antibodies were detected by dot blot assay and an ELISA using guinea pig tissue transglutaminase (gp-tTG) as antigen. The anti-endomysium antibodies were detected by an indirect immunofluorescence technique on cryostat sections of human umbilical cord. The anti-reticulin antibodies were also investigated by indirect immunofluorescence on cryostat sections of kidney, liver and stomach of rat. The anti-gliadin antibodies were determined by an ELISA. The sensitivity of an ELISA for the detection of anti-tissue transglutaminase antibodies was 86% in children and 87% in adults and the sensitivity of dot blot assay was 57% in children and 54% in adults. The specificity of an ELISA and dot blot for the detection for anti-tissue transglutaminase antibodies was, respectively, 96% and 88% lower than that of anti-endomysium antibodies (100%). The sensitivity of anti-gliadin antibodies was 97% in children and 91% in adults and their specificity was 85%. The sensitivity of anti-reticulin antibodies was 94% in children and 87% in adults. Their specificity was 100%. The sensitivity and specificity of an ELISA for the detection of anti-tissue transglutaminase antibodies were better than that of dot blot assay. However, this dot blot assay could screen four celiac patients who have not had anti-tissue transglutaminase antibodies by an ELISA. The sensitivity of anti-endomysium antibodies was better than that of anti-tissue transglutaminase antibodies, anti-reticulin antibodies and anti-gliadin antibodies but in children aged less than 2 years, the sensitivity of anti-gliadin antibodies was better than that of anti-tissue transglutaminase antibodies.