(32)P-postlabeling detection of DNA adducts in peripheral white blood cells of greenhouse floriculturists from western Liguria, Italy.
Pesticides are widely used in agriculture to enhance crop yields and to control disease vectors. Floriculturists work frequently in greenhouses and may be exposed to high levels of pesticides, which may result in adverse health effects. To evaluate the relationship between exposure to pesticides and DNA adduct formation in peripheral WBCs of Italian floriculturists, the nuclease P1 modification of a (32)P-postlabeling assay was used to analyze WBC DNA from floriculturists (n = 26) and matched controls (n = 22). DNA adduct-positive samples were more frequent in floriculturists (11/26; 42%) than in matched controls (2/22; 9%) (P < 0.01). Slightly higher frequencies of DNA adduct-positive samples were observed in floriculturists > or = 44 years of age (53%) and in female floriculturists (57%). Floricultural practice was found to be associated with a significantly higher DNA adduct-positive rate in WBCs (rate ratio, 5.12; 95% confidence interval, 1.1-23.7) after allowing for the effects of age and gender. These two latter covariates were not significantly associated with DNA adduct-positive rates. The quantitative levels of DNA adducts were significantly higher in floriculturists than in matched controls according to the Mann-Whitney nonparametric statistic (P = 0.0052). The median adduct level for positive samples among floriculturists was 1.5/10(8) bases. A specific, well-visible spot, named alpha adduct, was detected in 7 out of the 11 DNA adduct-positive samples from floriculturists but in none of the (22 + 20) referent samples (P = 0.0004). The presence of pesticide-related DNA adducts was confirmed clearly using the butanol extraction procedure. Six of 8 floriculturists and 0 of 10 referents were found positive with this method. The median adduct level for positive samples was 6.0/10(8) bases. Two strong spots close to the origin could be identified in all six positive floriculturists, using the butanol extraction procedure. No association between DNA adducts and use of specific pesticides was observed.


Hematogenous vertebral osteomyelitis.
Hematogenous vertebral osteomyelitis is an increasingly important consideration in the differential diagnosis of patients presenting with back pain even in the absence of fever. The incidence of this disease is increasing and physicians must maintain a high index of suspicion to make a prompt diagnosis. In patients with progressive neurologic deficits, urgent diagnosis is imperative, as the window for necessary surgical intervention is narrow. Magnetic resonance imaging is the preferred radiologic modality because of its sensitivity. Bone biopsy is the gold standard for diagnosis and is usually performed percutaneously using CT-guidance. Antimicrobial therapy of at least six weeks in duration is recommended and should be tailored to pathogens recovered from bone biopsy culture. With improved diagnostic techniques and prompt management, adverse outcomes associated with a delay in diagnosis can be avoided.


Optimization of Nonambulant Mass Casualty Decontamination Protocols as Part of an Initial or Specialist Operational Response to Chemical Incidents.
Objective: The UK's Initial Operational Response (IOR) is a new process for improving the survival of multiple casualties following a chemical, biological, radiological or nuclear incident. Whilst the introduction of IOR represents a patient-focused response for ambulant casualties, there is currently no provision for disrobe and dry decontamination of nonambulant casualties. Moreover, the current specialist operational response (SOR) protocol for nonambulant casualty decontamination (also referred to as "clinical decontamination") has not been subject to rigorous evaluation or development. Therefore, the aim of this study was to confirm the effectiveness of putatively optimized dry (IOR) and wet (SOR) protocols for nonambulant decontamination in human volunteers. Methods: Dry and wet decontamination protocols were objectively evaluated using human volunteers. Decontamination effectiveness was quantified by liquid chromatography-mass spectrometry analysis of the recovery of a chemical warfare agent simulant (methylsalicylate) from skin and hair of volunteers, with whole-body fluorescence imaging to quantify the skin distribution of residual simulant. Results: Both the dry and wet decontamination processes were rapid (3 and 4 min, respectively) and were effective in removing simulant from the hair and skin of volunteers, with no observable adverse effects related to skin surface spreading of contaminant. Conclusions: Further studies are required to assess the combined effectiveness of dry and wet decontamination under more realistic conditions and to develop appropriate operational procedures that ensure the safety of first responders.


Hybrid gap modes induced by fiber taper waveguides: application in spectroscopy of single solid-state emitters deposited on thin films.
We show, via simulations, that an optical fiber taper waveguide can be an efficient tool for photoluminescence and resonant, extinction spectroscopy of single emitters, such as molecules or colloidal quantum dots, deposited on the surface of a thin dielectric membrane. Placed over a high refractive index membrane, a tapered fiber waveguide induces the formation of hybrid mode waves, akin to dielectric slotted waveguide modes, that provide strong field confinement in the low index gap region. The availability of such gap-confined waves yields potentially high spontaneous emission enhancement factors (approximately 20), fluorescence collection efficiencies (approximately 23%), and transmission extinction (approximately 20%) levels. A factor of two improvement in fluorescence and extinction levels is predicted if the membrane is instead replaced with a suspended channel waveguide. Two configurations, for operation in the visible (approximately 600 nm) and near-infrared (approximately 1300 nm) spectral ranges are evaluated, presenting similar performances.


Linking Objects to Actions: Encoding of Target Object and Grasping Strategy in Primate Ventral Premotor Cortex.
Neural activity in ventral premotor cortex (PMv) has been associated with the process of matching perceived objects with the motor commands needed to grasp them. It remains unclear how PMv networks can flexibly link percepts of objects affording multiple grasp options into a final desired hand action. Here, we use a relational encoding approach to track the functional state of PMv neuronal ensembles in macaque monkeys through the process of passive viewing, grip planning, and grasping movement execution. We used objects affording multiple possible grip strategies. The task included separate instructed delay periods for object presentation and grip instruction. This approach allowed us to distinguish responses elicited by the visual presentation of the objects from those associated with selecting a given motor plan for grasping. We show that PMv continuously incorporates information related to object shape and grip strategy as it becomes available, revealing a transition from a set of ensemble states initially most closely related to objects, to a new set of ensemble patterns reflecting unique object-grip combinations. These results suggest that PMv dynamically combines percepts, gradually navigating toward activity patterns associated with specific volitional actions, rather than directly mapping perceptual object properties onto categorical grip representations. Our results support the idea that PMv is part of a network that dynamically computes motor plans from perceptual information. The present work demonstrates that the activity of groups of neurons in primate ventral premotor cortex reflects information related to visually presented objects, as well as the motor strategy used to grasp them, linking individual objects to multiple possible grips. PMv could provide useful control signals for neuroprosthetic assistive devices designed to interact with objects in a flexible way.


Surviving your new CEO.
Almost 50% of the largest American firms will have a new CEO within the next four years; your company could very well be next. Senior executives know that a CEO transition means they're in for a round of firings, organizational reshuffles, and other unwelcome career changes. When your career suddenly depends on the views of a person you may not know, how worried should you be? According to the authors--very. They investigated the 2002-2004 CEO turnover rates of the top 1,000 U.S. companies and interviewed more than a dozen CEOs, each of whom had taken over at least one very large organization. Their study reveals that when a new CEO takes charge, remaining top managers are more likely than not to be shown the door. Those who leave often land in a lower position at a new company, work in a much smaller firm, or retire altogether. The news is not all grim, however. The interviewees offer some pointers on how to create a good impression and maximize your chances of survival and success under the new regime. Some of that advice may surprise you. One CEO pointed out, for instance, that "managers do not realize how much the CEO is looking for teammates on day one. I am amazed at how few people come through the door and say, 'I want to help. I may not be perfect, but I buy into your vision:" Other recommendations are more intuitive, such as learning the new CEO's working style, understanding her agenda, and helping her look good in her new position by achieving positive operating results--and soon. Along with the inevitable stresses, the authors point out, CEO transitions can provide opportunities. Whether you reinvigorate your career within your company or find fulfillment elsewhere, the key lies in deciding what you want to do--and then doing it right.


Experimental reduction of intromittent organ length reduces male reproductive success in a bug.
It is now clear in many species that male and female genital evolution has been shaped by sexual selection. However, it has historically been difficult to confirm correlations between morphology and fitness, as genital traits are complex and manipulation tends to impair function significantly. In this study, we investigate the functional morphology of the elongate male intromittent organ (or processus) of the seed bug Lygaeus simulans, in two ways. We first use micro-computed tomography (micro-CT) and flash-freezing to reconstruct in high resolution the interaction between the male intromittent organ and the female internal reproductive anatomy during mating. We successfully trace the path of the male processus inside the female reproductive tract. We then confirm that male processus length influences sperm transfer by experimental ablation and show that males with shortened processi have significantly reduced post-copulatory reproductive success. Importantly, male insemination function is not affected by this manipulation per se. We thus present rare, direct experimental evidence that an internal genital trait functions to increase reproductive success and show that, with appropriate staining, micro-CT is an excellent tool for investigating the functional morphology of insect genitalia during copulation.


Effectiveness of blood donor questionnaire directed at risk factor for transfusion transmitted infections in Pakistani population.
Deferring blood donors who admit to high-risk behavior on questioning are likely to eliminate those in window period for transfusion transmitted infections (TTI). However, many questions have been implemented in some countries as part of donor history questionnaire, based on precautionary principle and not on evidence, and can result in increased donor losses. This study aims to identify effective risk-directed questions having high predictive value, in local context which can form part of blood donor deferral policies. For this, a case control study in a hospital blood bank having donation services was carried out prospectively over a period of three years. Two hundred and twenty donors, who were repeatedly reactive for HBsAg, anti-HCV, anti-HIV with EIA, and syphilis with TPHA, were the cases. Eight hundred and eighty four controls were the donors who tested negative for all TTI test. All donors answered seven hepatitis risk directed questions and their responses and reactivity status for TTI were used for statistical analysis with SPSS ver. 15. Positive predictive value for history of jaundice at any age for HBsAg was 20%, while PPV for history of surgery in previous six months for both HBsAg and anti-HCVHCV was also around 20%, based on pretest probability of 7%. The post-test probability for these questions was around 30%. Odds ratios with 95% CI did not reveal any significant association of hepatitis with any of seven questions. Donor losses after deferring on basis of two questions were 5.3% per year, while deferral rate after all seven questions was 20%. Donors should be permanently deferred if there is history of jaundice at any age, while deferral period after surgery should be one year. Other risk-directed questions should not be used to defer donors. Donor deferral policies should be evidence based and questions with proven efficacy should be made part of donor history questionnaire to minimize donor losses.


Effect of angiostatin on 1,2-dimethylhydrazine-induced colon cancer in mice.
Antiangiogenic therapy is supposed to be an attractive approach for antitumor treatment. Human plasminogen-derived angiostatin K1-3 is one of the most potent antiangiogenic agents known currently. However, it is unclear whether angiostatin has got protective effects on colon cancer. So we investigated the protective effects of angiostatin on 1,2-dimethylhydrazine (DMH)-induced colon cancer in mice. Thirty Balb/C male mice, weighing 25-30 g and 8 weeks of age, were used. Twenty of the mice were treated with DMH subcutaneously (20 mg/kg) once a week for 12 weeks. Six mice died during the DMH injection and surviving mice were divided into two groups (7 mice in DMH and 7 mice in DMH + angiostatin groups). In the angiostatin group, 6 weeks after the last DMH injection the animals were first treated with angiostatin (20 μg/mouse) intraperitoneally and then subcutaneously every 48 h (5 μg/mouse) throughout a period of 12 weeks. The animals were killed after 30 weeks for histopathological examination. When we look at the distribution of lesions in the colon, they mainly occurred in the distal colon. The incidence of mean colonic lesions in a tumor-bearing mouse was 9.85 ± 4.91 in those treated with DMH and 8.71 ± 3.49 in those treated with angiostatin. The incidence of colon tumors was not significantly affected by low dose of angiostatin, and we noticed that the number of lesions decreased by 12% in DMH + angiostatin group compared to the number of the lesions in DMH group, but this decrease was not statistically significant (p > 0.05). The administration period of angiostatin corresponds to the precancerous period and the reduction in the number of lesions could be important for the protective function of angiostatin in DMH + angiostain group. We assume that therapeutic effects of angiostatin are related to its doses, route of administration, frequency and administration period. In addition, we believe that combination of high doses of angiostatin with radiation, gene therapy or chemotherapy might be successful in proper tumor model.


Concurrent chemoradiotherapy with S-1 as first-line treatment for patients with oropharyngeal cancer.
S-1 is an oral fluoropyrimidine. The purpose of this study was to review the clinical outcomes of S-1 with concurrent radiotherapy for patients with oropharyngeal cancer. Between 2002 and 2007, 38 patients with oropharyngeal cancer treated concurrently with S-1 and definitive radiotherapy were reviewed. The clinical stage was Stage I in 4 patients, Stage II in 7, Stage III in 7, and Stage IV in 20. S-1 was administered orally twice daily for 4 consecutive weeks followed by a 2-week drug withdrawal. The initial dose of S-1 was 65 mg/m(2)/day. All patients were treated using three-dimensional conformal radiotherapy with a median total dose of 65.1 Gy (range, 60.0-71.0 Gy). Clinical outcomes and major acute toxicities were analyzed based on medical records and clinical follow-up. With a median follow-up time of 33 months, the 3-year estimates of local-regional control, distant metastases-free survival, disease-free survival, and overall survival for all patients were 75%, 80%, 65%, and 80%, respectively. The 3-year estimates of local-regional control according to stage were 100% for Stages I and II, 86% for Stage III, and 56% for Stage IV. The rate of ≥ Grade 3 acute mucositis was 32%, and the rate of ≥ Grade 3 hematological toxicities was 8%. No other severe toxicities were observed. Concurrent chemoradiotherapy with S-1 was found to be effective, especially for early disease. The treatment-related toxicities were acceptable, and the incidence of myelotoxicity was low. Further study must be carried out to compare with other chemotherapy regimens.


A mouse model reproducing the pathophysiology of neonatal group B streptococcal infection.
Group B streptococcal (GBS) meningitis remains a devastating disease. The absence of an animal model reproducing the natural infectious process has limited our understanding of the disease and, consequently, delayed the development of effective treatments. We describe here a mouse model in which bacteria are transmitted to the offspring from vaginally colonised pregnant females, the natural route of infection. We show that GBS strain BM110, belonging to the CC17 clonal complex, is more virulent in this vertical transmission model than the isogenic mutant BM110∆cylE, which is deprived of hemolysin/cytolysin. Pups exposed to the more virulent strain exhibit higher mortality rates and lung inflammation than those exposed to the attenuated strain. Moreover, pups that survive to BM110 infection present neurological developmental disability, revealed by impaired learning performance and memory in adulthood. The use of this new mouse model, that reproduces key steps of GBS infection in newborns, will promote a better understanding of the physiopathology of GBS-induced meningitis.


Moving towards IoT Based Digital Communication: An Efficient Utilization of Power Spectrum Density for Smart Cities.
The future of the Internet of Things (IoT) is interlinked with digital communication in smart cities. The digital signal power spectrum of smart IoT devices is greatly needed to provide communication support. The line codes play a significant role in data bit transmission in digital communication. The existing line-coding techniques are designed for traditional computing network technology and power spectrum density to translate data bits into a signal using various line code waveforms. The existing line-code techniques have multiple kinds of issues, such as the utilization of bandwidth, connection synchronization (CS), the direct current (DC) component, and power spectrum density (PSD). These highlighted issues are not adequate in IoT devices in smart cities due to their small size. However, there is a need to design an effective line-code method to deal with these issues in digital IoT-based communication for smart technologies, which enables smart services for smart cities. In this paper, the Shadow Encoding Scheme (SES) is proposed to transmit data bits efficiently by using a physical waveform in the smart cities' ecosystem. SES provides a reliable transmission over the physical medium without using extra bandwidth and with ideal PSD. In it, the shadow copy of the repeating bitstream is forwarded, rather than repeating the actual stream again and again. The PSD is calculated with the help of mathematical equations to validate SES. MATLAB simulator is used to simulate SES and compared with other well-known digital line-code techniques. The bit error rate is also compared between SES and the chirp spread spectrum (CSS) for the specific data frames. The coordinates of the PSD graph are also shown in tabular form, which shows a vivid picture of the working conditions of various line codes.


Jelly Belly trans-synaptic signaling to anaplastic lymphoma kinase regulates neurotransmission strength and synapse architecture.
In Drosophila, the secreted signaling molecule Jelly Belly (Jeb) activates anaplastic lymphoma kinase (Alk), a receptor tyrosine kinase, in multiple developmental and adult contexts. We have shown previously that Jeb and Alk are highly enriched at Drosophila synapses within the CNS neuropil and neuromuscular junction (NMJ) and postulated a conserved intercellular signaling function. At the embryonic and larval NMJ, Jeb is localized in the motor neuron presynaptic terminal whereas Alk is concentrated in the muscle postsynaptic domain surrounding boutons, consistent with anterograde trans-synaptic signaling. Here, we show that neurotransmission is regulated by Jeb secretion by functional inhibition of Jeb-Alk signaling. Jeb is a novel negative regulator of neuromuscular transmission. Reduction or inhibition of Alk function results in enhanced synaptic transmission. Activation of Alk conversely inhibits synaptic transmission. Restoration of wild-type postsynaptic Alk expression in Alk partial loss-of-function mutants rescues NMJ transmission phenotypes and confirms that postsynaptic Alk regulates NMJ transmission. The effects of impaired Alk signaling on neurotransmission are observed in the absence of associated changes in NMJ structure. Complete removal of Jeb in motor neurons, however, disrupts both presynaptic bouton architecture and postsynaptic differentiation. Nonphysiologic activation of Alk signaling also negatively regulates NMJ growth. Activation of Jeb-Alk signaling triggers the Ras-MAP kinase cascade in both pre- and postsynaptic compartments. These novel roles for Jeb-Alk signaling in the modulation of synaptic function and structure have potential implications for recently reported Alk functions in human addiction, retention of spatial memory, cognitive dysfunction in neurofibromatosis, and pathogenesis of amyotrophic lateral sclerosis.


Potential Mechanism for Some Postoperative C5 Palsies: An Anatomical Study.
Anatomical Study. Determine if shoulder depression (eg, taping the shoulders) might result in C5 nerve traction and subsequent injury. Postoperative C5 nerve palsy is a recognized entity that is still often enigmatic. Inferior shoulder depression is usually employed to assist with surgical visualization during cervical spine procedures. In the supine position, 10 adult fresh frozen human cadavers underwent dissection of the spinal cord and its adjacent dorsal, ventral roots, and spinal nerves from C4 to T1. In the supine position, the head was rotated ipsilaterally, contralaterally, and in lateral flexion. The shoulder was elevated, retracted, protracted, and depressed all with direct observation of nerve roots, intradural ventral/dorsal rootlets, or the spinal cord. The effects of these movements upon the cervical nerve rootlets were measured. The greatest displacement of nervous tissue was generated by shoulder depression and occurred primarily at the intradural rootlet level. The nerve rootlets that underwent the greatest average displacement were found at C5, with a decreasing gradient to C7 and no gross motion at C8 or T1. With maximal shoulder depression, C5-C7 rootlet tension produced cord movement to the ipsilateral side, touching the dura mater covering the lateral vertebral column with the C5 nerve root moving farthest. Shoulder depression is often used during cervical spine surgery. In cadavers, shoulder depression causes significant tension and displacement of the C5 nerve rootlets, and in the extreme, cord displacement to the ipsilateral side. This could be a mechanism for injury, putting patients at greater risk for postoperative C5 palsy. 5.


Patho-epigenetics: histone deacetylases as targets of pathogens and therapeutics.
In recent years, the interplay of epigenetics and infection moved into the limelight. Epigenetic regulation describes modifications in gene expression without alterations of the DNA sequence. In eukaryotes, this mechanism is central for fundamental cellular processes such as cell development and differentiation, but it is also involved in more specific tasks such as the response to infection by a pathogen. One of the most common types of epigenetic changes is the modification of histones. Histones, the small protein building blocks that are wrapped with DNA are the fundamental packaging unit of chromatin. Histones can be modified by linking different moieties to them-one of the most abundant ones is acetylation. Histone acetylation is regulated by two main classes of enzymes, histone acetyl transferases (HAT) and their counterparts, histone deacetylases (HDAC). Given the high abundance and importance in regulating gene expression, histone acetylation is an excellent target for pathogens to manipulate the host cell to their advantage. Targeting HDACs gained particular interest in recent years, due to the increased use of HDAC inhibitors in clinical practice. Recently, the possibility to fight an infection with HDAC inhibitors was suggested as an alternative to overcome the ever-growing problem of antibiotic resistance. In this review, we focus on the regulation of HDACs and their involvement in immune cell function. We then highlight different mechanisms employed by pathogens to manipulate histone deacetylases and we discuss the possibility of HDAC inhibitors as therapeutics to fight infections.


Thalamotomy as a treatment option for tremor after ineffective deep brain stimulation.
As the number of deep brain stimulation (DBS) surgeries increases, complications from malpositioned electrodes, tolerance to stimulation and loss of efficacy have also increased. Our objective was to assess thalamotomy as a salvage treatment option after ineffective DBS. A retrospective chart analysis of patients who underwent ipsilateral stereotactic thalamotomy after an ineffective ventrointermediate nucleus DBS procedure was undertaken. Patient outcome was based on follow-up visit chart notes, and a nonvalidated patient telephone questionnaire to assess patients' perception of tremor and functional ability after thalamotomy. Six patients with essential tremor and 1 with tremor-predominant Parkinson's disease met our inclusion criteria. Thalamotomies were undertaken for ineffective DBS due to dysarthria and paresthesias with programming in 2 patients, tremor that failed to respond to increased DBS despite a lack of side effects in 2 patients, malpositioned electrode in 2 patients, and sudden loss of DBS efficacy following eye surgery in 1 patient. Following thalamotomy, 3 patients reported improvement in symptoms and function, 3 patients reported improvement in symptoms that were not reflected in functional improvement, and 1 patient reported no improvement in symptoms or function. Thalamotomy may provide a viable salvage solution in patients who fail to respond to DBS due to complications such as malpositioned electrodes, tolerance to stimulation or loss of efficacy.


Complete biosynthesis of opioids in yeast.
Opioids are the primary drugs used in Western medicine for pain management and palliative care. Farming of opium poppies remains the sole source of these essential medicines, despite diverse market demands and uncertainty in crop yields due to weather, climate change, and pests. We engineered yeast to produce the selected opioid compounds thebaine and hydrocodone starting from sugar. All work was conducted in a laboratory that is permitted and secured for work with controlled substances. We combined enzyme discovery, enzyme engineering, and pathway and strain optimization to realize full opiate biosynthesis in yeast. The resulting opioid biosynthesis strains required the expression of 21 (thebaine) and 23 (hydrocodone) enzyme activities from plants, mammals, bacteria, and yeast itself. This is a proof of principle, and major hurdles remain before optimization and scale-up could be achieved. Open discussions of options for governing this technology are also needed in order to responsibly realize alternative supplies for these medically relevant compounds.


Modeling and computation of multistep batch testing for infectious diseases.
We propose a mathematical model based on probability theory to optimize COVID-19 testing by a multistep batch testing approach with variable batch sizes. This model and simulation tool dramatically increase the efficiency and efficacy of the tests in a large population at a low cost, particularly when the infection rate is low. The proposed method combines statistical modeling with numerical methods to solve nonlinear equations and obtain optimal batch sizes at each step of tests, with the flexibility to incorporate geographic and demographic information. In theory, this method substantially improves the false positive rate and positive predictive value as well. We also conducted a Monte Carlo simulation to verify this theory. Our simulation results show that our method significantly reduces the false negative rate. More accurate assessment can be made if the dilution effect or other practical factors are taken into consideration. The proposed method will be particularly useful for the early detection of infectious diseases and prevention of future pandemics. The proposed work will have broader impacts on medical testing for contagious diseases in general.


Model suggesting that replication of influenza virus is regulated by stabilization of replicative intermediates.
The RNA-dependent RNA polymerase of influenza A virus is responsible for both transcription and replication of negative-sense viral RNA. It is thought that a "switching" mechanism regulates the transition between these activities. We demonstrate that, in the presence of preexisting viral RNA polymerase and nucleoprotein (NP), influenza A virus synthesizes both mRNA (transcription) and cRNA (replication) early in infection. We suggest that there may be no switch regulating the initiation of RNA synthesis and present a model suggesting that nascent cRNA is degraded by host cell nucleases unless it is stabilized by newly synthesized viral RNA polymerase and NP.


Electron microscopic analysis reveals that replication factor C is sequestered by single-stranded DNA.
Replication factor C (RF-C) is a eukaryotic heteropentameric protein required for DNA replication and repair processes by loading proliferating cell nuclear antigen (PCNA) onto DNA in an ATP-dependent manner. Prior to loading PCNA, RF-C binds to DNA. This binding is thought to be restricted to a specific DNA structure, namely to a primer/template junction. Using the electron microscope we have examined the affinity of human heteropentameric RF-C and the DNA-binding region within the large subunit of RF-C from Drosophila melanogaster (dRF-Cp140) to heteroduplex DNA. The electron microscopic data indicate that both human heteropentameric RF-C and the DNA-binding region within dRF-Cp140 are sequestered by single-stranded DNA. No preferential affinity for the 3' or 5' transition points from single- to double-stranded DNA was evident.


The C-terminal dimerization motif of cyclase-associated protein is essential for actin monomer regulation.
Cyclase-associated protein (CAP) is a conserved actin-regulatory protein that functions together with actin depolymerizing factor (ADF)/cofilin to enhance actin filament dynamics. CAP has multiple functional domains, and the function to regulate actin monomers is carried out by its C-terminal half containing a Wiskott-Aldrich Syndrome protein homology 2 (WH2) domain, a CAP and X-linked retinitis pigmentosa 2 (CARP) domain, and a dimerization motif. WH2 and CARP are implicated in binding to actin monomers and important for enhancing filament turnover. However, the role of the dimerization motif is unknown. Here, we investigated the function of the dimerization motif of CAS-2, a CAP isoform in the nematode Caenorhabditis elegans, in actin monomer regulation. CAS-2 promotes ATP-dependent recycling of ADF/cofilin-bound actin monomers for polymerization by enhancing exchange of actin-bound nucleotides. The C-terminal half of CAS-2 (CAS-2C) has nearly as strong activity as full-length CAS-2. Maltose-binding protein (MBP)-tagged CAS-2C is a dimer. However, MBP-CAS-2C with a truncation of either one or two C-terminal β-strands is monomeric. Truncations of the dimerization motif in MBP-CAS-2C nearly completely abolish its activity to sequester actin monomers from polymerization and enhance nucleotide exchange on actin monomers. As a result, these CAS-2C variants, also in the context of full-length CAS-2, fail to compete with ADF/cofilin to release actin monomers for polymerization. CAS-2C variants lacking the dimerization motif exhibit enhanced binding to actin filaments, which is mediated by WH2. Taken together, these results suggest that the evolutionarily conserved dimerization motif of CAP is essential for its C-terminal region to exert the actin monomer-specific regulatory function.


Prognostic Value of Nodal Response After Preoperative Treatment of Gastric Adenocarcinoma.
Background: Pathologically positive lymph nodes (ypN+) after preoperative chemotherapy are associated with poor survival in patients with gastric cancer. Little is known about the association between response to preoperative therapy and the benefit of postoperative therapy. Methods: This retrospective cohort study of the National Cancer Database included patients with clinically node-positive (cN+) gastric cancer treated with preoperative therapy followed by surgery (2006-2014). Preoperative treatment modality was categorized as the inclusion of radiation therapy (RT) or chemotherapy alone. Pretreatment clinical and pathologic stages were used to determine pathologic treatment response rates. The association between overall risk of death and preoperative treatment, disease response, and adjuvant therapy use was evaluated using multivariable Cox regression. Results: Preoperative RT was used in 53.6% of 1,976 patients with cN+ gastric cancer, (74.3% cardia and 10.1% noncardia). The nodal response rate was 38.9% and was higher with RT than with chemotherapy alone (cardia, 46.0% vs 29.1%; P<.001; noncardia, 43.8% vs 31.9%; P=.06). Preoperative RT was associated with an approximate 2-fold increase in the odds of pathologic response compared with chemotherapy. Overall, use of adjuvant therapy was not associated with a decreased risk of death. A primary tumor response with residual nodal disease was not associated with survival (hazard ratio [HR], 1.03; 95% CI, 0.66-1.60). However, a nodal response with residual primary disease was significantly associated with survival (HR, 0.54; 95% CI, 0.44-0.65). Conclusions: More than one-third of node-positive gastric cancers showed pathologic nodal response with preoperative treatment. RT is associated with a higher response than chemotherapy. Patients with ypN+ disease have worse survival, regardless of whether they receive postoperative therapy. Future gastric cancer trials should evaluate the role of preoperative RT and individualize postoperative therapy use.


High Production of 2,3-butanediol by a Mutant Strain of the Newly Isolated Klebsiella pneumoniae SRP2 with Increased Tolerance Towards Glycerol.
Biodiesel, a renewable fuel produced by transesterification of animal fats and vegetable oils, generates about 10% (v/v) of crude glycerol as a core byproduct. The high volume of this non bio-degradable glycerol is becoming of a great environmental and economical concern due to its worldwide ever-growing surplus. Herein we report a high production of 2,3-butanediol (2,3-BD) from pure and biodiesel derived crude glycerol using a mutant K. pneumoniae SRM2 obtained from a newly isolated strain Klebsiella pneumoniae SRP2. The mutant strain SRM2 with standing high glycerol concentration (220 g L-1 of medium) could rapidly convert glycerol aerobically to 2,3-BD, a versatile product extensively used in chemical, pharmaceutical and fuel industries Our study revealed that an increased GDH activity led to a substantially enhanced production of 2,3-BD. The mutant strain exhibited 1.3-fold higher activity of GDH than that of parent strain (500.08 vs. 638.6 µmol min -1 mg -1 protein), yielding of 32.3 g L-1 and 77.5 g L-1 2,3-BD with glycerol in batch and fed-batch process respectively. However, in batch culture with crude glycerol, cell growth and glycerol consumption were expressively boosted, and 2,3-BD production was 27.7 g L-1 from 75.0 g/L crude glycerol. In this report, the optimal conditions for high production of 2,3-BD were defined in a completely aerobic process, and 0.59 g g-1 product yield of 2,3-BD was attained by the mutated strain K. pneumoniae SRM2, which is the highest amount obtained from batch biotransformation process of glycerol metabolism till today. These results indicated that our newly developed mutant can tolerate high concentration of glycerol, have a high glycerol utilization rate, and high product yield of 2,3-BD. It is demonstrated that the mutant strain K. pneumoniae SRM2 has an ability to produce fewer co-products at trace concentrations at higher glycerol concentrations, and could be a potential candidate for 2,3-DB production in an industrial bioconversion process.


A controlled trial of two nucleoside analogues plus indinavir in persons with human immunodeficiency virus infection and CD4 cell counts of 200 per cubic millimeter or less. AIDS Clinical Trials Group 320 Study Team.
The efficacy and safety of adding a protease inhibitor to two nucleoside analogues to treat human immunodeficiency virus type 1 (HIV-1) infection are not clear. We compared treatment with the protease inhibitor indinavir in addition to zidovudine and lamivudine with treatment with the two nucleosides alone in HIV-infected adults previously treated with zidovudine. A total of 1156 patients not previously treated with lamivudine or protease inhibitors were stratified according to CD4 cell count (50 or fewer vs. 51 to 200 cells per cubic millimeter) and randomly assigned to one of two daily regimens: 600 mg of zidovudine (or stavudine) and 300 mg of lamivudine, or that regimen with 2400 mg of indinavir. The primary end point was the time to the development of the acquired immunodeficiency syndrome (AIDS) or death. The proportion of patients whose disease progressed to AIDS or death was lower with indinavir, zidovudine, and lamivudine (6 percent) than with zidovudine and lamivudine alone (11 percent; estimated hazard ratio, 0.50; 95 percent confidence interval, 0.33 to 0.76; P=0.001). Mortality in the two groups was 1.4 percent and 3.1 percent, respectively (estimated hazard ratio, 0.43; 95 percent confidence interval, 0.19 to 0.99; P=0.04). The effects of treatment were similar in both CD4 cell strata. The responses of CD4 cells and plasma HIV-1 RNA paralleled the clinical results. Treatment with indinavir, zidovudine, and lamivudine as compared with zidovudine and lamivudine alone significantly slows the progression of HIV-1 disease in patients with 200 CD4 cells or fewer per cubic millimeter and prior exposure to zidovudine.


Mortality associated with atrial fibrillation in patients with myocardial infarction: a systematic review and meta-analysis.
Atrial fibrillation (AF) is a common finding in patients with myocardial infarction (MI). Atrial fibrillation is not generally perceived by clinicians as a critical event during the acute phase of MI; however, its prognostic influence in MI remains controversial. Furthermore, contradictory data exist concerning the risk of death according to AF timing. This article, a systematic review and first meta-analysis, aims to quantify the mortality risk associated with AF in MI patients and its timing. A comprehensive search of several electronic databases (1970 to 2010; adults, any language) identified MI studies that evaluated mortality related to AF. Evidence was reviewed by 2 blinded reviewers with a formal assessment of the methodological quality of the studies. Adjusted odds ratios were pooled across studies using the random-effects model. The I(2) statistic was used to assess heterogeneity. In the 43 included studies (278 854 subjects), the mortality odds ratio associated with AF was 1.46 (95% confidence interval, 1.35 to 1.58; I(2)=76%; 23 studies). This worse prognosis persisted regardless of the timing of AF; the odds ratio of mortality for new AF with no prior history of AF was 1.37 (95% confidence interval, 1.26 to 1.49), I(2)=28%, 9 studies), and for prior AF was 1.28 (95% confidence interval, 1.16 to 1.40; I(2)=24%; 4 studies). The sensitivity analysis of new AF studies adjusting for confounding factors did not show a decrease in risk of death. Atrial fibrillation is associated with increased risk of mortality in MI patients. New AF with no history of AF before MI remained associated with an increased risk of mortality even after adjustment for several important AF risk factors. These subsequent increases in mortality suggest that AF can no longer be considered a nonsevere event during MI.


Cyclooxygenase-2 directly induces MCF-7 breast tumor cells to develop into exponentially growing, highly angiogenic and regionally invasive human ductal carcinoma xenografts.
Based on our studies demonstrating first time evidence that the cyclooxygenase-2 (Cox-2) enzyme is abundant within invasive human breast tumors, we developed a clonally derived human breast tumor cell clone designated as MCF-7/Cox-2 Clone 10 by transfection of human Cox-2 cDNA into slow growing, Cox-2 null, non-metastatic MCF-7 human breast tumor cells. The present studies evaluated the biological characteristics of the MCF-7/Cox-2 Clone 10 human breast tumors compared to the characteristics of MCF-7/empty vector control tumors when grown in vivo following injection of 5x10(6) tumor cells into mammary fat pads of ovariectomized female Crl:Nu-Foxn1(nu) mice implanted with slow release 17-beta estradiol pellets. At 60 days after tumor cell injection, MCF-7/Cox-2 Clone 10 human breast tumors were 4-fold greater (p < 0.01) in volume than MCF-7/empty vector control tumors. MCF-7/Cox-2 Clone 10 human breast tumor xenografts were highly angiogenic based on histological observation of large-bore blood vessels, which was confirmed by immunohistochemical staining with anti-CD-31 antibody and quantitation of mean vessel density. MCF-7/Cox-2 Clone 10 human breast tumor cells were present within regional lymph nodes adjacent to mammary fat pads with their local invasion confirmed by Western blotting of Cox-2-protein. This unique Cox-2-dependent breast tumor model rapidly produces large, angiogenic, locally invasive human breast tumor xenografts in mammary fat pads of ovariectomized female Crl:Nu-Foxn1(nu) mice at 42-60 days which recapitulate human breast ductal carcinomas. This unique model may be invaluable as a means to evaluate preclinical safety and efficacy of novel adjuvant therapies for women with metastastic breast cancer including prostanoid receptor antagonists, newly developed anti-angiogenic therapies, as well as other novel approaches for targeting and destruction of human breast tumors and their vasculature.


Perceived skills for sports performance after primary hip arthroplasty: a cross-sectional study.
Recommendations arising from existing literature regarding restrictions and benefits of sporting activities after joint replacement surgery vary widely. As hip arthroplasty patients are becoming increasingly active, their expectations about post-operative function are constantly evolving. The aim of this study is to identify the perception of patients regarding their performance in sports activities after hip arthroplasty. This cross-sectional study included all patients undergoing primary hip arthroplasty, for any diagnosis, between January 2009 and January 2016. By applying a telephone survey, practice of sports before surgery, resumption after surgery, level of performance, and causes of non-resumption of sports activities were assessed. Data of 531 patients were obtained. Of these, 13% were engaged in sports before surgery. The most frequently practiced sports were golf (27.5%) and tennis (22%). Of the 72 patients that practiced sports, only 44.4% (30 patients) returned to this activity after surgery. Nonetheless, 71% of these patients reported to have an equal or better athletic performance than before surgery. The main causes reported by patients not to return to sports were the fear of injury and recommendation of the surgeon. A significant number of patients return to sports after hip arthroplasty and most of them perceive a good athletic performance after surgery. These findings should enrich the pre-operative assessment of patient's expectations, particularly for those who wish to resume physical activity.


Mast cell tryptase and chymase reverse airway smooth muscle relaxation induced by vasoactive intestinal peptide in the ferret.
Recent evidence suggests that nonadrenergic airway relaxation may be controlled by vasoactive intestinal peptide (VIP). The magnitude and duration of smooth muscle relaxation in response to VIP may be influenced by rates of peptide degradation after release from efferent peptidergic neurons. To explore the potential role of mast cell mediators in modulating neural control of airway tone, we studied the effect of the mast cell proteases tryptase and chymase on airway smooth muscle relaxation induced by VIP in ferret airway. Tracheal rings precontracted by serotonin (10(-6) M) in a muscle bath were relaxed by VIP (10(-7) M). We found that protease-rich supernatant obtained by degranulation of dog mastocytoma cells reversed VIP-induced relaxation, as did highly purified tryptase and chymase incubated with the tracheal rings. Either enzyme completely reversed the effect of VIP, but tryptase was more potent than chymase, paralleling previous test tube observations on the relative rates of VIP cleavage by the two enzymes. Inhibitors of mast cell tryptase and chymase preincubated with the supernatant or with the purified proteases prevented reversal of VIP-induced relaxation. Mast cell proteases did not reverse the tracheal relaxation caused by the nonpeptide adrenergic agonist isoproterenol. These findings show that mast cell proteases tryptase and chymase counteract the smooth muscle relaxant effects of VIP in ferret trachea and suggest a potential role for the mast cell proteases in the modulation of nonadrenergic neural control of airway tone by VIP.


Gastric outlet and duodenal obstruction from inflammatory pancreatic disease.
During a ten-year period, 16 patients with gastric outlet and duodenal obstruction due to inflammatory pancreatic disease were seen. The cause of obstruction was chronic pancreatitis in ten patients, pseudocysts with associated pancreatitis in five patients, and pancreatic abscess in one patient. All patients had nausea and vomiting, 14 had abdominal pain, and five had weight loss greater than 4.5 kg. Diagnosis was made by plain abdominal film in one case, upper gastrointestinal tract roentgenographic series in 15 cases, and endoscopy in 11 cases. Mobilization of the duodenum relieved the obstruction in two patients. Fixed obstruction remained in 14 patients. This was relieved by gastrojejunostomy in 12 patients. Gastrojejunostomy was combined with drainage of a pseudocyst in three patients, a dilated pancreatic duct in three patients, and a dilated common bile duct in four patients. Obstruction was relieved by pseudocyst drainage in two patients. Associated common duct and pancreatic duct obstruction must be identified preoperatively.


Overshadowing: a silver lining to a dark cloud in horse training.
Overshadowing is a process known in behavioral science that occurs when two stimuli of different strengths are applied simultaneously to a nonhuman animal. Typically, the stronger stimulus overshadows the weaker one, resulting in attenuation of the weaker stimulus. This phenomenon explains ways in which the decreased responsiveness and consequent conflict behaviors (and possibly learned helplessness and wastage) in some performance horses can result from the application of two concurrent aversive stimuli. Despite some adverse consequences in the context of ridden horses, overshadowing can have serendipitous benefits because it offers an efficient method of desensitization for certain stimuli that are sometimes highly aversive: the saddle/girth pressure, clippers, aerosols, and needles. Desensitization with concurrent overshadowing appears to be comparatively rapid, particularly with highly aversive stimuli, possibly because attentional mechanisms are diverted to the more salient stimulus. It is important to note that, following the overshadowing procedure, the effects appear to be retained when assessed on subsequent days. Using 4 examples, this article presents a preliminary exploration of the beneficial use of a poorly understood, underutilized--yet promising--phenomenon that warrants further investigation.


Purification and characterization of human DNA damage checkpoint Rad complexes.
Checkpoint Rad proteins function early in the DNA damage checkpoint signaling cascade to arrest cell cycle progression in response to DNA damage. This checkpoint ensures the transmission of an intact genetic complement to daughter cells. To learn about the damage sensor function of the human checkpoint Rad proteins, we purified a heteropentameric complex composed of hRad17-RFCp36-RFCp37-RFCp38-RFCp40 (hRad17-RFC) and a heterotrimeric complex composed of hRad9-hHus1-hRad1 (checkpoint 9-1-1 complex). hRad17-RFC binds to DNA, with a preference for primed DNA and possesses weak ATPase activity that is stimulated by primed DNA and single-stranded DNA. hRad17-RFC forms a complex with the 9-1-1 heterotrimer reminiscent of the replication factor C/proliferating cell nuclear antigen clamp loader/sliding clamp complex of the replication machinery. These findings constitute biochemical support for models regarding the roles of checkpoint Rads as damage sensors in the DNA damage checkpoint response of human cells.


Estimated risks of radiation-induced fatal cancer from pediatric CT.
In light of the rapidly increasing frequency of pediatric CT examinations, the purpose of our study was to assess the lifetime cancer mortality risks attributable to radiation from pediatric CT. Organ doses as a function of age-at-diagnosis were estimated for common CT examinations, and estimated attributable lifetime cancer mortality risks (per unit dose) for different organ sites were applied. Standard models that assume a linear extrapolation of risks from intermediate to low doses were applied. On the basis of current standard practice, the same exposures (milliampere-seconds) were assumed, independent of age. The larger doses and increased lifetime radiation risks in children produce a sharp increase, relative to adults, in estimated risk from CT. Estimated lifetime cancer mortality risks attributable to the radiation exposure from a CT in a 1-year-old are 0.18% (abdominal) and 0.07% (head)-an order of magnitude higher than for adults-although those figures still represent a small increase in cancer mortality over the natrual background rate. In the United States, of approximately 600,000 abdominal and head CT examinations annually performed in children under the age of 15 years, a rough estimate is that 500 of these individuals might ultimately die from cancer attributable to the CT radiation. The best available risk estimates suggest that pediatric CT will result in significantly increased lifetime radiation risk over adult CT, both because of the increased dose per milliampere-second, and the increased lifetime risk per unit dose. Lower milliampere-second settings can be used for children without significant loss of information. Although the risk-benefit balance is still strongly tilted toward benefit, because the frequency of pediatric CT examinations is rapidly increasing, estimates that quantitative lifetime radiation risks for children undergoing CT are not negligible may stimulate more active reduction of CT exposure settings in pediatric patients.


The use of medicinal plants by primates: A missing link?
There is growing evidence that some species of wild nonhuman primate, especially chimpanzees, take herbal and clay medicines to treat and prevent disease. Such a primate pharmacopoeia may be a missing link in our understanding of the relationship between primate foraging and ranging strategies and plant chemistry; not all plant secondary compounds may be deleterious to the consumer. Just as study of traditional herbal medicines has yielded powerful drugs, so primate medicines may hint at drugs useful in treating human disease.


Hepatic steatosis and HIV infection.
There is increasing concern that patients with chronic HIV infection may be at increased risk of nonalcoholic fatty liver disease (NAFLD), which can evolve into nonalcoholic steatohepatitis (NASH) and cirrhosis. Multiple factors have been hypothesized to be necessary for the development and progression of this condition. Potential risk factors, which tend to accumulate in the HIV-positive population, include metabolic derangements, chronic inflammation, hepatitis coinfection, and treatment with certain nucleoside reverse transcriptase inhibitors (NRTIs). HIV-associated conditions such as hyperlactatemia and lipodystrophy frequently overlap with fatty liver disease. The cornerstone of management of HIV-associated fatty liver disease is currently to treat the predominant underlying condition. There is a need for more epidemiologic data to better define the role of comorbidities and drugs in the development of NAFLD. Further work is also needed to elucidate the pathogenesis and to evaluate the therapeutic effect of treating comorbidities and avoiding certain antiretroviral drugs.


Inelastic collapse in simple shear flow of a granular medium.
Previous investigations have shown that inelastic collapse is a common feature of inelastic, hard-sphere simulations of nondriven (or unforced) flows, provided that the coefficient of restitution is small enough. The focus of the current effort is on a driven system, namely, simple shear flow. Two-dimensional, hard-sphere simulations have been carried out over a considerable range of restitution coefficients (r), solids fractions (nu), and numbers of particles (N). The results indicate that inelastic collapse is an integral feature of the sheared system. Similar to nondriven systems, this phenomenon is characterized by a string of particles engaging in numerous, repeated collisions just prior to collapse. The collapsed string is typically oriented along a 135 degrees angle from the streamwise direction. Inelastic collapse is also found to be more likely in systems with lower r, higher nu, and higher N, as is true for unforced systems. Nonetheless, an examination of the boundary between the collapsed and noncollapsed states reveals that the sheared system is generally more "resistant" to inelastic collapse than its nondriven counterpart. Furthermore, a dimensionless number V* is identified that represents the magnitude of the initial fluctuating velocities relative to that of a characteristic steady-state velocity (i.e., the product of shear rate and particle diameter). For values of V*>>O(1), the transient portion of the simulation is found to be more reminiscent of a nondriven system (i.e., isotropic particle bunching is observed instead of diagonal particle bands).


In vitro inhibition and induction of human hepatic cytochrome P450 enzymes by modafinil.
The ability of modafinil to affect human hepatic cytochrome P450 (CYP) activities was examined in vitro. The potential for inhibition of CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4/5, and CYP4A9/11 by modafinil (5-250 microM) was evaluated with pooled human liver microsomes. Modafinil exhibited minimal capacity to inhibit any CYP enzyme, except CYP2C19. Modafinil inhibited the 4'-hydroxylation of S-mephenytoin, a marker substrate for CYP2C19, reversibly and competitively with a K(i) value of 39 microM, which approximates the steady-state C(max) value of modafinil in human plasma at a dosage of 400 mg/day. No irreversible inhibition of any CYP enzyme was observed, and there was no evidence of metabolism-dependent inhibition. The potential for induction of CYP activity was evaluated by exposing primary cultures of human hepatocytes to modafinil (10-300 microM). Microsomes were then prepared and assayed for CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4/5 activities. The mean activities of microsomal CYP1A2, CYP2B6, and CYP3A4/5 from modafinil-treated hepatocytes were higher (up to 2-fold) than those in the solvent-treated controls but were less than those produced by reference inducers of these enzymes. At high concentrations of modafinil (>/=100 microM), the mean activity of CYP2C9 was decreased (up to 60%) relative to that in the solvent controls. Overall, modafinil was shown to have effects on human hepatic CYP1A2, CYP2B6, CYP2C9, CYP2C19, and CYP3A4/5 activities in vitro. Although effects obtained in vitro are not always predictive of effects in vivo, such results provide a rational basis for understanding drug-drug interactions that are observed clinically and for planning subsequent investigations.


Parvalbumin-expressing interneurons coordinate hippocampal network dynamics required for memory consolidation.
Activity in hippocampal area CA1 is essential for consolidating episodic memories, but it is unclear how CA1 activity patterns drive memory formation. We find that in the hours following single-trial contextual fear conditioning (CFC), fast-spiking interneurons (which typically express parvalbumin (PV)) show greater firing coherence with CA1 network oscillations. Post-CFC inhibition of PV+ interneurons blocks fear memory consolidation. This effect is associated with loss of two network changes associated with normal consolidation: (1) augmented sleep-associated delta (0.5-4 Hz), theta (4-12 Hz) and ripple (150-250 Hz) oscillations; and (2) stabilization of CA1 neurons' functional connectivity patterns. Rhythmic activation of PV+ interneurons increases CA1 network coherence and leads to a sustained increase in the strength and stability of functional connections between neurons. Our results suggest that immediately following learning, PV+ interneurons drive CA1 oscillations and reactivation of CA1 ensembles, which directly promotes network plasticity and long-term memory formation.


Clinical Predictors for Repeat Hospitalizations in Left Ventricular Assist Device (LVAD) Patients With Gastrointestinal Bleeding.
Patients implanted with left ventricular assist devices (LVAD) carry an increased risk of gastrointestinal bleeding (GIB), estimated at 25% in most studies. Significant efforts are employed in localizing and stopping the source of bleeding, but the rates of repeat hospitalization for GIB remain surprisingly high. Given the increasing incidence of LVAD-dependent end-stage heart failure and the excessive costs associated with repetitive endoscopic investigations, risk factors associated with re-bleeding need to be determined. The aim of our study was to investigate clinical predictors associated with repeat hospitalizations for GIB in patients implanted with a LVAD. We conducted a retrospective cohort using the prospectively assembled ventricular assist device database at the University of Alabama at Birmingham. We identified all end-stage heart failure patients who were implanted with a continuous-flow (CF) LVAD between Jan 1, 2009 and Dec 31, 2013. We excluded pulsatile devices, biventricular assist devices (BiVADs), right ventricular assist devices (RVADs), and patients under 19 years of age. There were 102 patients implanted with a CF-LVAD within the specified time period. With an average follow-up of 127 weeks, 32 (31.4%) patients developed GIB requiring 79 separate hospitalizations. Average time from LVAD implantation to first bleed was 343 days. The re-bleeding rate requiring readmission was 56.3% in those admitted with GIB, with eight (25%) of the patients necessitating multiple readmissions. The average hospital stay for a primary diagnosis of GIB was 9.45 days. Totally, 68 (86%) patients required endoscopic evaluation during their hospitalization, with 35 (44%) necessitating multiple procedures during the same admission. The average time to first endoscopy was 2.5 days with a median of 2 days. Patients receiving early endoscopy (< 48 h from admission) were 57% less likely to require future readmission for GIB compared to patients undergoing late endoscopy (> 48 h) (OR: 0.43, CI: 0.19 - 0.9). Other factors associated with repeat admissions for GIB included indication for LVAD (bridge to transplant had OR: 0.07, CI: 0.02 - 0.27), male gender (OR: 10.4, CI: 1.8 - 59), length of initial hospital stay (OR: 0.83, CI: 0.71 - 0.97), and INR on admission (OR: 3.6, CI: 1.46 - 8.8). Although not statistically significant, patients undergoing subsequent endoscopies during a single admission were 84% less likely to develop re-bleeding in the future (OR: 0.158, CI: 0.025 - 1.02). GIB in LVAD patients is a significant problem with high rates of readmission despite extensive endoscopic investigations and anticoagulant adjustments. Our experience revealed that early endoscopy, longer initial hospital stay, and better INR control were all associated with decreased rates of readmission for GIB in this population. These modifiable factors should be emphasized and addressed in the future to reduce the burdens associated with repeated hospitalizations.


Blunted Dopamine Transmission in Addiction: Potential Mechanisms and Implications for Behavior.
Positron emission tomography (PET) imaging consistently shows blunted striatal dopamine release and decreased dopamine D2 receptor availability in addiction. Here, we review the preclinical and clinical studies indicating that this neurobiological phenotype is likely to be both a consequence of chronic drug consumption and a vulnerability factor in the development of addiction. We propose that, behaviorally, blunted striatal dopamine transmission could reflect the increased impulsivity and altered cost/benefit computations that are associated with addiction. The factors that influence blunted striatal dopamine transmission in addiction are unknown. Herein, we give an overview of various factors, genetic, environmental, and social, that are known to affect dopamine transmission and that have been associated with the vulnerability to develop addiction. Altogether, these data suggest that blunted dopamine transmission and decreased D2 receptor availability are biomarkers both for the development of addiction and resistance to treatment. These findings support the view that blunted dopamine reflects impulsive behavior and deficits in motivation, which lead to the escalation of drug use.


Calibrating the end-Permian mass extinction.
The end-Permian mass extinction was the most severe biodiversity crisis in Earth history. To better constrain the timing, and ultimately the causes of this event, we collected a suite of geochronologic, isotopic, and biostratigraphic data on several well-preserved sedimentary sections in South China. High-precision U-Pb dating reveals that the extinction peak occurred just before 252.28 ± 0.08 million years ago, after a decline of 2 per mil (‰) in δ(13)C over 90,000 years, and coincided with a δ(13)C excursion of -5‰ that is estimated to have lasted ≤20,000 years. The extinction interval was less than 200,000 years and synchronous in marine and terrestrial realms; associated charcoal-rich and soot-bearing layers indicate widespread wildfires on land. A massive release of thermogenic carbon dioxide and/or methane may have caused the catastrophic extinction.


Quantifying the transfer and settling in NMR experiments with sample shuttling.
Nuclear magnetic resonance (NMR) in combination with pulsed magnetic field gradients has proven very successful for measuring molecular diffusion, where the correlation time of the motion is much shorter than the timescale of the experiment. In this article, it is demonstrated that a single-scan NMR technique to measure molecular diffusion can be employed to also study incoherent random motions over macroscopic length scales that show correlation times similar to the timescale of the experiment. Such motions are observed, for example, after the mixing of two components or after transferring a sample from one container into another. To measure the fluid settling, a series of magnetization helices were encoded onto a sample. Stimulated gradient echo trains were then generated after different mixing times, which enabled the determination of an effective dispersion coefficient for the fluid. This technique was used to optimize the timing of NMR experiments combined with dissolution dynamic nuclear polarization, where a sample was shuttled between two magnets. In addition to the decay of fluid turbulences, the presence of microbubbles in the sample tube at the end of the shuttling step was identified as another contribution to the NMR linewidth. Microbubbles could be indirectly observed through the line broadening effect on the NMR signal due to their different susceptibility compared to the solvent, which induced field gradients near the interfaces. Using these data, the signal attenuation caused by sample motion in single-scan two-dimensional correlation spectroscopy NMR experiments could be predicted with reasonable accuracy.


Antiepiligrin (laminin 5) cicatricial pemphigoid associated with an underlying gastric carcinoma producing laminin 5.
Although bullous pemphigoid and cicatricial pemphigoid are sometimes associated with malignancy, it remains uncertain whether such an association is pathogenetically related or just a coincidence attributable to the advanced age of the patients. We report a 61-year-old patient with antiepiligrin (laminin 5) cicatricial pemphigoid (AeCP) associated with an advanced gastric carcinoma. The gastric carcinoma cells in this patient were shown to produce laminin 5 by immunofluorescence microscopy, and the patient's serum contained autoantibodies directed against laminin 5 on immunoprecipitation. Furthermore, the blistering symptoms and the titre of antibasement membrane zone antibodies coordinately changed with the resection and subsequent relapse of the gastric cancer. These observations suggest that the gastric carcinoma producing laminin 5 may have induced the production of autoantibodies to this laminin, which were pathogenic to the skin and mucous membranes in this patient. This report demonstrates a link between this autoimmune subepithelial blistering disease and malignancy. It is of interest and potential great importance to examine other cases of AeCP for such a potential association.


Successful treatment of renal artery embolism even forty-eight hours after event.
Renal artery embolism is a disease that is easily missed due to its infrequent and nonspecific presentations. Although early diagnosis and optimal thrombolytic treatment can sometimes restore renal function, therapeutic guidelines have not yet been established. However, early anticoagulant therapy is beneficial and selective infusion of lytic agents into renal artery has been reported with increasing frequency and efficacy if used in the early stage. We report that intra-arterial thrombolytic therapy with low dose of 35 mg recombinant tissue plasminogen activator (t-PA) may be an effective and safe strategy for the treatment of renal artery embolism, despite the period of ischemia being longer than 48 hours.


Presentation patterns in women with pelvic venous disorders differ based on age of presentation.
The prevalence and presentation patterns in women with pelvic venous disorders (PeVD) secondary to pelvic venous insufficiency (PVI) at various ages are ill-defined. The purpose of this investigation was to determine if the types of symptoms, interventions, and treatment outcomes of women with PeVD varied with age progression. From January 2015 to December 2019, we retrospectively reviewed prospectively collected data on 1,280 women with PeVD from our electronic medical record at the Center for Vascular Medicine (CVM). Medical and surgical comorbidities, past medical history, presenting pelvic and lower extremity symptoms, Clinical, Etiology, Anatomy, Pathophysiology (CEAP) class, revised Venous Clinical Severity Score (rVCSS), visual analog pain score (VAS) and types of interventions were assessed. Patients were grouped into five categories based on age of initial presentation: 20-29, 30-39, 40-49, 50-59, and greater than or equal to 60. Patients were also subcategorized according to their course of treatment: Iliac venous stenting alone, ovarian vein embolization (OVE) alone, simultaneous iliac vein stenting and ovarian vein embolization, and staged iliac vein stenting and ovarian vein embolization. Differences in groups were analyzed utilizing chi square, analysis of variance and regression analysis with Graphpad Prism 8 (San Diego, CA) and SAS Studio 3.8 (Cary, NC) statistical software. From January 2015 through December 2019, 1,280 women were treated for PeVD. The average ages in each group were the following: 26.53 ± 2.90 (n = 57), 35.80 ± 2.84 (n = 238), 44.98 ± 2.78 (n = 345), 54.67 ± 2.90 (n = 324) and 68.39 ± 8.44 (n = 316) respectively. The prevalence of PVI by age group was 4.45%,18.59%, 26.95%, 25.31% and 24.70% respectively (p < 0.05). The prevalence of diabetes, hypertension, coronary artery disease (CAD) and hypercholesterolemia differed between age groups (p < 0.05). Significant differences in the prevalence of pelvic pain, dyspareunia, dysmenorrhea, vulvar varices, leg pain, leg edema, leg heaviness, restless legs, varicose veins, and leg ulcers were observed across treatment groups (p < 0.05). CEAP distribution differed by age with CEAP 0-2 decreasing with age, CEAP 3 progressively increasing with age and CEAP 4-6 predominantly observed in patients older than 50. Analysis of variance indicated that with each decile increase in age, the prevalence of leg symptoms increased (p < 0.05) with a reciprocal decrease in pelvic symptoms (p < 0.05). All intervention types were infrequent in twenty year old's. There was a linear increase in stenting with each decile increase in age. The prevalence of OVE was similar across all age groups. There was an overall improvement in VAS scores for all treatment and age groups. For patients in their 20 s, an initial improvement in VAS scores was observed with moderate recurrence of pain observed after three months. PeVD presents as a spectrum of signs and symptoms, with pelvic and leg symptoms being inversely related according to age. The prevalence of PeVD is lowest in patients in their twenties with differences in presentation observed with increasing age. Venous stenting progressively increases with each decile of age whereas the prevalence of OVE is similar regardless of age. There is overall improvement in symptoms post intervention, although women in their 20 s do not respond as well to intervention as women in other age groups. Future investigations will focus on determining which pelvic venous lesion is the predominant factor that needs correction to achieve maximal pain reduction.


Improved estimates of the radiation absorbed dose to the urinary bladder wall.
Specific absorbed fractions (SAFs) have been calculated as a function of the content in the urinary bladder in order to allow more realistic calculations of the absorbed dose to the bladder wall. The SAFs were calculated using the urinary bladder anatomy from the ICRP male and female adult reference computational phantoms. The urinary bladder and its content were approximated by a sphere with a wall of constant mass, where the thickness of the wall depended on the amount of urine in the bladder. SAFs were calculated for males and females with 17 different urinary bladder volumes from 10 to 800 mL, using the Monte Carlo computer program MCNP5, at 25 energies of mono-energetic photons and electrons ranging from 10 KeV to 10 MeV. The decay was assumed to be homogeneously distributed in the urinary bladder content and the urinary bladder wall, and the mean absorbed dose to the urinary bladder wall was calculated. The Monte Carlo simulations were validated against measurements made with thermoluminescent dosimeters. The SAFs obtained for a urine volume of 200 mL were compared to the values calculated for the urinary bladder wall using the adult reference computational phantoms. The mean absorbed dose to the urinary wall from (18)F-FDG was found to be 77 µGy/MBq formales and 86 µGy/MBq for females, while for (99m)Tc-DTPA the mean absorbed doses were 80 µGy/MBq for males and 86 µGy/MBq for females. Compared to calculations using a constant value of the SAF from the adult reference computational phantoms, the mean absorbed doses to the bladder wall were 60% higher for (18)F-FDG and 30% higher for (99m)Tc-DTPA using the new SAFs.


Diabetes induced changes in rat mesenchymal stem cells.
Diabetes mellitus, the single most important cause of vascular disease in the industrialized world, is also associated with bone loss and impaired fracture healing. Mesenchymal stem cells (MSCs) have the potential to differentiate into osteoblasts, chondrocytes and adipocytes and other mesenchymal cells and play a central role in bone formation and repair. Because of this, we have investigated the possibility that diabetes has direct effects on MSCs in vivo and that this might represent a cellular basis for diabetes-induced osteoporosis. We isolated MSCs from rats with streptozotocin-induced diabetes and analysed them ex vivo for their ability to proliferate and differentiate in the fibroblastic colony-forming unit assay. Effects of diabetes on bone metabolism in vivo were determined by analysing tibiae from control and diabetic animals by quantitative computerized tomography. The total number of colonies and osteoblastic colonies staining positive for alkaline phosphatase were quantified and both colony size and number were found to be significantly reduced in diabetic rats. The changes appear to be mediated by the induction of apoptosis and senescence by advanced glycation end products (AGEs), together with an increase in the receptor for AGEs (RAGE). These changes were paralleled by extensive loss of trabecular bone in the tibiae of the diabetic animals. These data suggest that MSCs become exhausted during diabetes and lose their differentiation potential, leading to a net loss of trabecular bone. Therefore, direct effects on MSCs may be responsible for some of the orthopaedic effects associated with diabetes.


miR-216a promotes breast cancer cell apoptosis by targeting PKCα.
Breast cancer (BC) is the most common cause of death in women throughout the world. MicroRNAs (miRNAs, miR) have been identified as key regulators in carcinogenesis of several cancers, including BC. MicroRNA-216a (miR-216a) is downregulated in several cancers. Here, we evaluated the effects of miRNA-216a on breast cancer cells and the underlying mechanisms. miR-216a level was quantified by real-time RT-PCR. Cell viability was analyzed by MTT assay. Wound-healing assay was performed for detection of cell migration. Apoptosis was detected by TUNEL and caspase-3 activity assay. Moreover, the level of protein expression was determined by Western blot. We found that miR-216a expression was remarkably decreased in both human BC tissues and MCF-7 cells. miR-216a overexpression dramatically suppressed the migration and promoted the apoptosis in cultured MCF-7 cells. We validated PKCα (protein kinase C alpha, PRKCA) as a direct target of miR-216a. Knockdown of PKCα induced apoptosis and inhibited migration in cultured MCF-7 cells which were reversed by miR-216a inhibitor. Moreover, the level of miR-216a is negatively correlated with PKCα in cell lines. Our results collectively suggest that miR-216a suppressed migration and promoted apoptosis in breast cancer cells by targeting PKCα. These findings indicate that manipulation of miR-216a expression may represent a novel therapeutic strategy in the treatment of breast cancer.


Isolation of a full-length human WNT7A gene implicated in limb development and cell transformation, and mapping to chromosome 3p25.
The Wnt gene family has a role in development as well as tumourigenesis. One mouse member, Wnt7a, is vital for limb development in vivo and also possesses transforming ability in vitro. This study reports the isolation of a full length of human homologue of mouse Wnt7a gene by library screening. Yeast artificial chromosome-fluorescence in situ hybridisation (YAC-FISH) mapped the WNT7A gene to chromosome 3p25. Human WNT7A had an ORF encoding a deduced protein of 349 aa that exhibited 97% and 92% identity to mouse Wnt7a at the aa and nucleic acid levels, respectively. It possessed the 22 conserved cysteine residues and 3 more at the amino terminus, and a putative poly A tail. This is the fifth human WNT gene in which a complete cDNA sequence had been determined.


Deciphering human heat shock transcription factor 1 regulation via post-translational modification in yeast.
Heat shock transcription factor 1 (HSF1) plays an important role in the cellular response to proteotoxic stresses. Under normal growth conditions HSF1 is repressed as an inactive monomer in part through post-translation modifications that include protein acetylation, sumoylation and phosphorylation. Upon exposure to stress HSF1 homotrimerizes, accumulates in nucleus, binds DNA, becomes hyper-phosphorylated and activates the expression of stress response genes. While HSF1 and the mechanisms that regulate its activity have been studied for over two decades, our understanding of HSF1 regulation remains incomplete. As previous studies have shown that HSF1 and the heat shock response promoter element (HSE) are generally structurally conserved from yeast to metazoans, we have made use of the genetically tractable budding yeast as a facile assay system to further understand the mechanisms that regulate human HSF1 through phosphorylation of serine 303. We show that when human HSF1 is expressed in yeast its phosphorylation at S303 is promoted by the MAP-kinase Slt2 independent of a priming event at S307 previously believed to be a prerequisite. Furthermore, we show that phosphorylation at S303 in yeast and mammalian cells occurs independent of GSK3, the kinase primarily thought to be responsible for S303 phosphorylation. Lastly, while previous studies have suggested that S303 phosphorylation represses HSF1-dependent transactivation, we now show that S303 phosphorylation also represses HSF1 multimerization in both yeast and mammalian cells. Taken together, these studies suggest that yeast cells will be a powerful experimental tool for deciphering aspects of human HSF1 regulation by post-translational modifications.


Guided mode biosensor based on grating coupled porous silicon waveguide.
Porous silicon waveguide biosensors that utilize grating couplers etched directly into porous silicon are demonstrated for improved molecular detection capabilities. Molecules are infiltrated through the grating couplers into the waveguide where they can interact with a guided waveguide mode. Hybridization of nucleic acids inside the waveguide is shown to significantly perturb the wave vector of the guided mode and is detected through angle-resolved reflectance measurements. A detection sensitivity of 7.3°/mM is demonstrated with selectivity better than 6:1 compared to mismatched sequences. Experimental results are in good agreement with calculations based on rigorous coupled wave analysis. Use of the all-porous silicon grating-coupled waveguide allows improved interaction of the optical field with surface-bound molecules compared to evanescent wave-based biosensors.


Repair of oxidative DNA damage in gram-positive bacteria: the Lactococcus lactis Fpg protein.
The formamidopyrimidine DNA glycosylase gene (fpg-L) of the Gram-positive microaerophilic bacterium Lactococcus lactis subsp. cremoris ML3 has been cloned, characterized and sequenced. The fpg-L gene is composed of 819 bp encoding a protein of 31.3 kDa (Fpg-L). The deduced amino acid sequence of the Fpg-L protein shows 59% similarity and 38% identity with the Escherichia coli Fpg protein (Fpg-E). Polyclonal antibodies against Fpg-E react with the Fpg-L protein. The Fpg-L protein was purified to apparent homogeneity from the overproducing E. coli strain BH410 hosting plasmid pVE1064, which carries fpg-L under the control of the E. coli lac promoter. In its active form, Fpg-L is a 30 kDa monomeric enzyme with a measured isoelectric point of 9.0. It contains one zinc per molecule and has a zinc finger motif localized at the carboxyterminal end (Cys-X2-Cys-X16-Cys-X2-Cys-X3-COOH). The Fpg-L protein has two enzyme activities: DNA glycosylase, which excises 2,6-diamino-4-hydroxy-5N-methylformamidopyrimidine and 7,8-dihydro-8-oxoguanine, and DNA nicking at abasic sites. Furthermore, the expression of the fpg-L gene in fpg and mutY mutants of E. coli suppresses their spontaneous GC-->TA mutator phenotype. The similarity of the activity of the two Fpg proteins and its conversation in evolutionarily distant bacteria may reflect the importance of its role in protecting bacterial DNA against oxidative free radicals.


Unconditional regard buffers children's negative self-feelings.
Unconditional regard refers to the feeling that one is accepted and valued by others without conditions. Psychological theory suggests that experiences of unconditional regard lead children to feel that they are valuable despite setbacks. We hypothesized that reflecting on experiences of unconditional regard would buffer children's negative self-feelings (eg, shame, insecurity, powerlessness) in the face of setbacks. To test this hypothesis, we randomized children to reflect on experiences of unconditional regard or other experiences, and examined their response to an academic setback 3 weeks later. Participants (11-15 years old) were randomly assigned to reflect for 15 minutes on experiences of unconditional regard (n = 91), conditional regard (n = 80), or other social experiences (n = 76). Research personnel, teachers, and classmates remained blind to condition assignment. Three weeks later, after receiving their course grades, children reported their self-feelings. Course grades were obtained from school records. Receiving low course grades represents a salient and painful real-world setback for children. Replicating previous research, children who received lower grades experienced more negative self-feelings (P < .001). As predicted, this well-established relationship was significantly attenuated among children who had reflected, 3 weeks previously, on experiences of unconditional regard (Ps < .03). Reflecting on unconditional regard specifically reduced negative self-feelings after low grades (P = .01), not after average or high grades (Ps > .17). Reflecting on unconditional regard buffered children's selves against the adverse impact of an academic setback over an extended period of time. Unconditional regard may thus be an important psychological lever to reduce negative self-feelings in youth.


Phylogeography and genetic ancestry of tigers (Panthera tigris).
Eight traditional subspecies of tiger (Panthera tigris),of which three recently became extinct, are commonly recognized on the basis of geographic isolation and morphological characteristics. To investigate the species' evolutionary history and to establish objective methods for subspecies recognition, voucher specimens of blood, skin, hair, and/or skin biopsies from 134 tigers with verified geographic origins or heritage across the whole distribution range were examined for three molecular markers: (1) 4.0 kb of mitochondrial DNA (mtDNA) sequence; (2) allele variation in the nuclear major histocompatibility complex class II DRB gene; and (3) composite nuclear microsatellite genotypes based on 30 loci. Relatively low genetic variation with mtDNA,DRB,and microsatellite loci was found, but significant population subdivision was nonetheless apparent among five living subspecies. In addition, a distinct partition of the Indochinese subspecies P. t. corbetti in to northern Indochinese and Malayan Peninsula populations was discovered. Population genetic structure would suggest recognition of six taxonomic units or subspecies: (1) Amur tiger P. t. altaica; (2) northern Indochinese tiger P. t. corbetti; (3) South China tiger P. t. amoyensis; (4) Malayan tiger P. t. jacksoni, named for the tiger conservationist Peter Jackson; (5) Sumatran tiger P. t. sumatrae; and (6) Bengal tiger P. t. tigris. The proposed South China tiger lineage is tentative due to limited sampling. The age of the most recent common ancestor for tiger mtDNA was estimated to be 72,000-108,000 y, relatively younger than some other Panthera species. A combination of population expansions, reduced gene flow, and genetic drift following the last genetic diminution, and the recent anthropogenic range contraction, have led to the distinct genetic partitions. These results provide an explicit basis for subspecies recognition and will lead to the improved management and conservation of these recently isolated but distinct geographic populations of tigers.


Non invasive assessment of liver fibrosis.
Liver fibrosis is the final common pathway of most chronic liver diseases of various etiologies.The development of fibrosis determines the management policy as well as the prognosis. In recent years it became clear that the fibrotic process is reversible. Therefore early detection of liver fibrosis is critical. At present, liver biopsy is considered the gold standard for the detection of fibrosis. Nevertheless liver biopsy is an invasive procedure limiting both its acceptance by patients, and their compliance for repeated procedures. More over it is a questionable gold standard due to significant sample error and intra and inter observer variability. These limitations have led to the development of noninvasive techniques for assessing the presence and the degree of liver fibrosis, such as biomarkers and transient elastography. The accuracy of these methods as compared to liver biopsy, as well as their use and limitations are reviewed in the article.


ConcatSeq: A method for increasing throughput of single molecule sequencing by concatenating short DNA fragments.
Single molecule sequencing (SMS) platforms enable base sequences to be read directly from individual strands of DNA in real-time. Though capable of long read lengths, SMS platforms currently suffer from low throughput compared to competing short-read sequencing technologies. Here, we present a novel strategy for sequencing library preparation, dubbed ConcatSeq, which increases the throughput of SMS platforms by generating long concatenated templates from pools of short DNA molecules. We demonstrate adaptation of this technique to two target enrichment workflows, commonly used for oncology applications, and feasibility using PacBio single molecule real-time (SMRT) technology. Our approach is capable of increasing the sequencing throughput of the PacBio RSII platform by more than five-fold, while maintaining the ability to correctly call allele frequencies of known single nucleotide variants. ConcatSeq provides a versatile new sample preparation tool for long-read sequencing technologies.


Assessment of MR angiography versus arteriography for evaluation of cervical carotid bifurcation disease.
To assess the accuracy of MR angiography alone in screening for vascular stenosis of the common carotid bifurcation. Two hundred two common carotid bifurcations in 101 patients were evaluated with MR angiography and selective contrast arteriography. A two-dimensional time-of-flight pulse sequence was used to obtain sequential transverse images through the common carotid bifurcations. These images were reprojected with a maximum intensity pixel ray-tracing algorithm. Both examinations were blindly graded as either normal or mildly stenotic (0%-29%), moderately stenotic (30%-69%), severely stenotic (70%-99%), or occluded. Of the 202 common carotid bifurcations, 119 were classified as normal-mild stenosis by contrast arteriography. In this category, MR angiography correctly identified 114 of these as normal to mild stenosis. Among 21 common carotid bifurcations graded as moderate stenosis by arteriography, 15 were correctly graded as moderate by MR angiography. Among 45 common carotid bifurcations graded as severe stenosis by arteriography, 41 of these were correctly graded as severe by MR angiography. There were 17 complete occlusions which were all correctly graded by MR angiography. MR angiography with its high rate of agreement with contrast arteriography can be regarded as an accurate screening method of the common carotid bifurcation.


RA and FGF signalling are required in the zebrafish otic vesicle to pattern and maintain ventral otic identities.
During development of the zebrafish inner ear, regional patterning in the ventral half of the otic vesicle establishes zones of gene expression that correspond to neurogenic, sensory and non-neural cell fates. FGF and Retinoic acid (RA) signalling from surrounding tissues are known to have an early role in otic placode induction and otic axial patterning, but how external signalling cues are translated into intrinsic patterning during otic vesicle (OV) stages is not yet understood. FGF and RA signalling pathway members are expressed in and around the OV, suggesting important roles in later patterning or maintenance events. We have analysed the temporal requirement of FGF and RA signalling for otic development at stages after initial anteroposterior patterning has occurred. We show that high level FGF signalling acts to restrict sensory fates, whereas low levels favour sensory hair cell development; in addition, FGF is both required and sufficient to promote the expression of the non-neural marker otx1b in the OV. RA signalling has opposite roles: it promotes sensory fates, and restricts otx1b expression and the development of non-neural fates. This is surprisingly different from the earlier requirement for RA signalling in specification of non-neural fates via tbx1 expression, and highlights the shift in regulation that takes place between otic placode and vesicle stages in zebrafish. Both FGF and RA signalling are required for the development of the otic neurogenic domain and the generation of otic neuroblasts. In addition, our results indicate that FGF and RA signalling act in a feedback loop in the anterior OV, crucial for pattern refinement.


Cerebellar Volume and Executive Function in Parkinson Disease with and without Freezing of Gait.
Freezing of gait (FOG) affects approximately 50% of people with Parkinson Disease (PD), impacting quality of life and placing financial and emotional strain on the individual and caregivers. People with PD and FOG have similar deficits in motor adaptation and cognition as individuals with cerebellar lesions, indicating the cerebellum may play a role in FOG. To examine potential differences in cerebellar volumes and their relationships with cognition between PD with (FOG+) and without FOG (FOG-). Sixty-three participants were divided into two groups, FOG+ (n = 25) and FOG- (n = 38), based on the New Freezing of Gait Questionnaire. Cognitive assessment included Trail Making, Stroop, Verbal Fluency, and Go-NoGo executive function tasks. All participants completed structural T1- and T2-weighted MRI scans. Imaging data were processed with FreeSurfer and the Spatially Unbiased Infratentorial toolbox to segment the cerebellum into individual lobules. FOG+ performed significantly worse on phonemic verbal fluency (F(1, 22) = 7.06, p = 0.01) as well as the Go-NoGo task (F(1, 22) = 9.00, p = 0.004). We found no differences in cerebellar volumes between groups (F(4, 55) = 1.42, p = 0.24), but there were significant relationships between verbal fluency measures and lobule volumes in FOG-. These findings underscore the need for longitudinal studies to better characterize potential changes in cerebellar volume, cognitive function, and functional connectivity between people with PD with and without FOG.


Oncogenic Gata1 causes stage-specific megakaryocyte differentiation delay.
The megakaryocyte/erythroid Transient Myeloproliferative Disorder (TMD) in newborns with Down Syndrome (DS) occurs when N-terminal truncating mutations of the hemopoietic transcription factor GATA1, that produce GATA1short protein (GATA1s), are acquired early in development. Prior work has shown that murine GATA1s, by itself, causes a transient yolk sac myeloproliferative disorder. However, it is unclear where in the hemopoietic cellular hierarchy GATA1s exerts its effects to produce this myeloproliferative state. Here, through a detailed examination of hemopoiesis from murine GATA1s ES cells and GATA1s embryos we define defects in erythroid and megakaryocytic differentiation that occur relatively late in hemopoiesis. GATA1s causes an arrest late in erythroid differentiation in vivo, and even more profoundly in ES-cell derived cultures, with a marked reduction of Ter-119 cells and reduced erythroid gene expression. In megakaryopoiesis, GATA1s causes a differentiation delay at a specific stage, with accumulation of immature, kit-expressing CD41hi megakaryocytic cells. In this specific megakaryocytic compartment, there are increased numbers of GATA1s cells in S-phase of cell cycle and reduced number of apoptotic cells compared to GATA1 cells in the same cell compartment. There is also a delay in maturation of these immature GATA1s megakaryocytic lineage cells compared to GATA1 cells at the same stage of differentiation. Finally, even when GATA1s megakaryocytic cells mature, they mature aberrantly with altered megakaryocyte-specific gene expression and activity of the mature megakaryocyte enzyme, acetylcholinesterase. These studies pinpoint the hemopoietic compartment where GATA1s megakaryocyte myeloproliferation occurs, defining where molecular studies should now be focussed to understand the oncogenic action of GATA1s.


Chromoplasts--the last stages in plastid development.
The results of investigations on the development of chromoplast fine structures in various plants are reviewed. Emphasis is placed on the specific pigment-containing structures and their development during chromoplast formation. There is a large variety of these structures, although four fundamental types can be discerned. These are plastoglobules, membranes, crystals, and tubules. During chromoplast development, various types of structure follow one after the other, or they may even be present simultaneously in the same chromoplast. Depending on the structures present in chromoplasts their pigment content also varies. It is still not clear whether the type of structure defines the pigment content of the chromoplast or vice-versa. Various possible ways of chromoplast development and dedifferentiation are discussed.


TMS evidence for the involvement of the right occipital face area in early face processing.
Extensive research has demonstrated that several specialized cortical regions respond preferentially to faces. One such region, located in the inferior occipital gyrus, has been dubbed the occipital face area (OFA). The OFA is the first stage in two influential face-processing models, both of which suggest that it constructs an initial representation of a face, but how and when it does so remains unclear. The present study revealed that repetitive transcranial magnetic stimulation (rTMS) targeted at the right OFA (rOFA) disrupted accurate discrimination of face parts but had no effect on the discrimination of spacing between these parts. rTMS to left OFA had no effect. A matched part and spacing discrimination task that used house stimuli showed no impairment. In a second experiment, rTMS to rOFA replicated the face-part impairment but did not produce the same effect in an adjacent area, the lateral occipital cortex. A third experiment delivered double pulses of TMS separated by 40 ms at six periods after stimulus presentation during face-part discrimination. Accuracy dropped when pulses were delivered at 60 and 100 ms only. These findings indicate that the rOFA processes face-part information at an early stage in the face-processing stream.


Self-Assembly of Aβ40, Aβ42 and Aβ43 Peptides in Aqueous Mixtures of Fluorinated Alcohols.
Fluorinated alcohols such as hexafluoroisopropanol (HFIP) and trifluoroethanol (TFE) have the ability to promote α-helix and β-hairpin structure in proteins and peptides. HFIP has been used extensively to dissolve various amyloidogenic proteins and peptides including Aβ, in order to ensure their monomeric status. In this paper, we have investigated the self-assembly of Aβ40, Aβ42, and Aβ43 in aqueous mixtures of fluorinated alcohols from freshly dissolved stock solutions in HFIP. We have observed that formation of fibrillar and non-fibrillar structures are dependent on the solvent composition. Peptides form fibrils with ease when reconstituted in deionized water from freshly dissolved HFIP stocks. In aqueous mixtures of fluorinated alcohols, either predominant fibrillar structures or clustered aggregates were observed. Aqueous mixtures of 20% HFIP are more favourable for Aβ fibril formation as compared to 20% TFE. When Aβ40, Aβ42, and Aβ43 stocks in HFIP are diluted in 50% aqueous mixtures in phosphate buffer or deionized water followed by slow evaporation of HFIP, Aβ peptides form fibrils in phosphate buffer and deionized water. The clustered structures could be off-pathway aggregates. Aβ40, Aβ42, and Aβ43 showed significant α-helical content in freshly dissolved HFIP stocks. The α-helical conformational intermediate in Aβ40, Aβ42, and Aβ43 could favour the formation of both fibrillar and non-fibrillar aggregates depending on solvent conditions and rate of α-helical to β-sheet transition.