Treatment Burden in People with Hypertension is Correlated with Patient Experience with Self-Management.
New hypertension guidelines in the United States may require more people to take multiple medications or implement lifestyle changes. Increased treatment burden may be an unintended consequence and lead to worse health outcomes. Our study examined whether treatment burden is associated with factors related to self-management in those with hypertension. We conducted a cross-sectional mailed survey of patients from 2 medical centers in Minnesota. Participants with 2 or more medical conditions completed the Patient Experience with Treatment and Self-management (PETS), a validated treatment burden questionnaire, as well as measures of confidence in self-management ability, health literacy, health care-related financial difficulties, and perception of provider interpersonal skills. We used partial correlation analyses, controlling for age, sex, race, and education, to test relationships among study variables. Of 254 respondents who had a diagnosis of hypertension, 54% were female, 74% were non-Hispanic White, and the mean age was 67 years. People with hypertension who reported having lower confidence in self-management ability, lower health literacy, more financial difficulties, and health care providers with poorer interpersonal skills reported higher treatment burden scores (PETS scales correlation magnitude range 0.09 to 0.62, P < .05 on all but 4). The strongest associations were observed for medical information and physical/mental exhaustion with self-management (correlation magnitudes from 0.25 to 0.54, P < .01). Hypertension treatment guideline stringent blood pressure criteria may lead to more interventions-medical or lifestyle-creating strains on populations already challenged by abstract disease self-management and at risk of experiencing disparities in cardiovascular health outcomes.


Influence of soot carbon on the soil-air partitioning of polycyclic aromatic hydrocarbons.
Soil-air partitioning is one of the key processes controlling the regional and global cycling and storage of polycyclic aromatic hydrocarbons (PAHs). However, the specific processes dominating the partitioning of PAHs between these two environmental compartments still need to be elucidated. Stable and distinct atmospheric conditions paralleling different soil properties are found at Tenerife island (28 degrees 18'N, 16 degrees 29'W), which is located in permanent inversion layer conditions, and they provide interesting model cases for the study of air-soil partitioning. Analysis of phenanthrene, pyrene, fluoranthene, and chrysene showed concentrations 4- to 10-fold higher below than above the inversion layer. Similarly, soil total organic carbon (TOC) and black carbon (BC) were 11 and 3 times higher, respectively, below the inversion layer than above. The octanol-air partition coefficient (K(OA)) derived model provides a good description of PAH soil-air partitioning coefficients (K(P)) below the inversion layer but underpredicts them in the area dominated by deposition of long-range transported aerosols without inputs of organic matter from local vegetation. Inclusion of soot carbon in the soil-air partitioning model results in good agreement between predicted and measured K(P) in this area but in overpredicted K(P) values for the soils under the vegetation cover, which shows that the influence of soil soot carbon on PAH air-soil partitioning depends on its abundance relative to soil organic carbon. Absorption into organic matter is the dominant process in soils containing high organic carbon concentrations, whereas adsorption onto soot carbon becomes relevant in soils with low organic carbon and high soot content.


Enhancing Intelligence: From the Group to the Individual.
Research aimed at testing whether short-term training programs can enhance intelligence is mainly concentrated on behavior. Expected positive effects are found sometimes, but the evidence is far from conclusive. It is assumed that training must evoke changes in the brain for observing genuine improvements in behavior. However, behavioral and brain data are seldom combined in the same study. Here we present one example of this latter type of research summarizing, discussing, and integrating already published results. The training program was based on the adaptive dual n-back task, and participants completed a comprehensive battery measuring fluid and crystallized ability, along with working memory and attention control, before and after training. They were also submitted to MRI scanning at baseline and post-training. Behavioral results revealed positive effects for visuospatial processing across cognitive domains. Brain imaging data were analyzed by longitudinal voxel-based morphometry, tensor-based morphometry, surface-based morphometry, and structural connectivity. The integration of these multimodal brain results provides clues about those observed in behavior. Our findings, along with previous research and current technological advances, are considered from the perspective that we now live in ideal times for (a) moving from the group to the individual and (b) developing personalized training programs.


Effects of halothane on GABAergic and glutamatergic transmission in isolated hippocampal nerve-synapse preparations.
We evaluated the effects of halothane on synaptic and extrasynaptic GABA(A) and glutamate receptor responses using mechanically dissociated rat hippocampal CA3 neurons in which the well isolated neurons retain functional native nerve endings (the 'synaptic bouton' preparation). The preparation allows the simultaneous comparison of extrasynaptic GABA(A) and glutamate receptors, activated by bath applied GABA and glutamate, respectively, to the synaptic receptors measured as spontaneous and evoked postsynaptic currents. Paired-pulse synaptic responses evoked by focal electrical stimulation were also measured to evaluate any presynaptic effects. Halothane enhanced the extrasynaptic GABA(A)-receptor mediated postsynaptic responses in a concentration dependent fashion. At clinically relevant concentrations, halothane significantly increased both the amplitude and frequency of spontaneous postsynaptic inhibitory currents (sIPSCs) mediated by synaptic GABA(A) receptors. The relative amplitude of evoked IPSCs (eIPSCs) was also increased, concurrent with a decrease in failure rate and a significantly decreased eIPSC paired-pulse ratio. Halothane concentration dependently decreased the extrasynaptic glutamate-receptor induced postsynaptic responses but had no effects on spontaneous or evoked excitatory postsynaptic currents. These results suggest that halothane acts predominantly at presynaptic sites at GABAergic synapses to enhance inhibitory transmission at CA3 synapses, although it also increases extra-synaptic GABA responses. At excitatory synapses on to CA3 neurons, halothane has no presynaptic action-effecting only extrasynaptic receptors. Our results have clarified the locus of effects of the volatile anesthetic halothane at excitatory and inhibitory synapses, drawing somewhat different conclusions from those deduced from slices and culture systems.


Efficient modification of λ-DNA substrates for single-molecule studies.
Single-molecule studies of protein-nucleic acid interactions frequently require site-specific modification of long DNA substrates. The bacteriophage λ is a convenient source of high quality long (48.5 kb) DNA. However, introducing specific sequences, tertiary structures, and chemical modifications into λ-DNA remains technically challenging. Most current approaches rely on multi-step ligations with low yields and incomplete products. Here, we describe a molecular toolkit for rapid preparation of modified λ-DNA. A set of PCR cassettes facilitates the introduction of recombinant DNA sequences into the λ-phage genome with 90-100% yield. Extrahelical structures and chemical modifications can be inserted at user-defined sites via an improved nicking enzyme-based strategy. As a proof-of-principle, we explore the interactions of S. cerevisiae Proliferating Cell Nuclear Antigen (yPCNA) with modified DNA sequences and structures incorporated within λ-DNA. Our results demonstrate that S. cerevisiae Replication Factor C (yRFC) can load yPCNA onto 5'-ssDNA flaps, (CAG)13 triplet repeats, and homoduplex DNA. However, yPCNA remains trapped on the (CAG)13 structure, confirming a proposed mechanism for triplet repeat expansion. We anticipate that this molecular toolbox will be broadly useful for other studies that require site-specific modification of long DNA substrates.


Feasibility of endovascular recanalization for symptomatic cervical internal carotid artery occlusion.
This study sought to report technical details and clinical results of the first series of endovascular recanalization for cervical internal carotid artery (ICA) occlusion. Cervical ICA occlusion is associated with impaired cerebral perfusion, which may lead to ischemic cerebral symptoms and hemodynamic infarcts. Neither surgical nor endovascular revascularization has been shown to benefit this population. Endovascular recanalization was attempted in 30 patients with ICA occlusions (27 men; age 72.1 +/- 8.0 years, range 48 to 85 years). Recurrent neurologic deficit or cerebral ischemia by perfusion study, after known ICA occlusion, was noted in all patients. Strategies and devices for coronary occlusion intervention were applied, including microcatheter-supported tapered-tip stiff coronary guidewires. Contralateral ICA stenosis was found in 9 patients (30%). All patients underwent independent neurologic and duplex ultrasound follow-up. The overall technical success rate was 73% (22 of 30). No neck hematoma, intracranial hemorrhage, or hyperperfusion was noted. One (3.3%) fatal brainstem infarction occurred 1 day after a successful ICA procedure, with angiographically proven acute basilar artery occlusion and patent ICA stent. Baseline ophthalmic artery flow was reversed in 15 of the 22 successfully recanalized patients, and was normalized in 12 after the procedure. There was no new cerebral ischemic event or neurologic death for a mean follow-up of 16.1 +/- 18.5 months. Endovascular recanalization for cervical ICA occlusion is feasible with acceptable midterm clinical results.


The complete sequence of the group-specific antigen, VP7, of African horsesickness disease virus serotype 4 reveals a close relationship to bluetongue virus.
The complete sequence of the S7 RNA that codes for the major group-specific coat protein. VP7, of African horsesickness virus serotype 4 (AHSV-4) was determined from cDNA analyses and found to be 1179 nucleotides in length. One single open reading frame of 353 codons was observed defining a protein of Mr 38,107 with a net charge of -1.5 at neutral pH. Comparison of the AHSV-4 VP7 sequence with that of bluetongue virus serotype 10 revealed an overall similarity of 44%, with the amino- and carboxy-terminal regions exhibiting the greatest levels of homology. In addition, potential secondary structures of the terminal sequences of the S7 RNA segments of AHSV-4 and BTV serotypes 10, 13 and 17 are presented.


Scene Configuration and Object Reliability Affect the Use of Allocentric Information for Memory-Guided Reaching.
Previous research has shown that egocentric and allocentric information is used for coding target locations for memory-guided reaching movements. Especially, task-relevance determines the use of objects as allocentric cues. Here, we investigated the influence of scene configuration and object reliability as a function of task-relevance on allocentric coding for memory-guided reaching. For that purpose, we presented participants images of a naturalistic breakfast scene with five objects on a table and six objects in the background. Six of these objects served as potential reach-targets (= task-relevant objects). Participants explored the scene and after a short delay, a test scene appeared with one of the task-relevant objects missing, indicating the location of the reach target. After the test scene vanished, participants performed a memory-guided reaching movement toward the target location. Besides removing one object from the test scene, we also shifted the remaining task-relevant and/or task-irrelevant objects left- or rightwards either coherently in the same direction or incoherently in opposite directions. By varying object coherence, we manipulated the reliability of task-relevant and task-irrelevant objects in the scene. In order to examine the influence of scene configuration (distributed vs. grouped arrangement of task-relevant objects) on allocentric coding, we compared the present data with our previously published data set (Klinghammer et al., 2015). We found that reaching errors systematically deviated in the direction of object shifts, but only when the objects were task-relevant and their reliability was high. However, this effect was substantially reduced when task-relevant objects were distributed across the scene leading to a larger target-cue distance compared to a grouped configuration. No deviations of reach endpoints were observed in conditions with shifts of only task-irrelevant objects or with low object reliability irrespective of task-relevancy. Moreover, when solely task-relevant objects were shifted incoherently, the variability of reaching endpoints increased compared to coherent shifts of task-relevant objects. Our results suggest that the use of allocentric information for coding targets for memory-guided reaching depends on the scene configuration, in particular the average distance of the reach target to task-relevant objects, and the reliability of task-relevant allocentric information.


Hypoperfusion of brain parenchyma is associated with the severity of chronic cerebrospinal venous insufficiency in patients with multiple sclerosis: a cross-sectional preliminary report.
Several studies have reported hypoperfusion of the brain parenchyma in multiple sclerosis (MS) patients. We hypothesized a possible relationship between abnormal perfusion in MS and hampered venous outflow at the extracranial level, a condition possibly associated with MS and known as chronic cerebrospinal venous insufficiency (CCSVI). We investigated the relationship between CCSVI and cerebral perfusion in 16 CCSVI MS patients and 8 age- and sex-matched healthy controls. Subjects were scanned in a 3-T scanner using dynamic susceptibility, contrast-enhanced, perfusion-weighted imaging. Cerebral blood flow (CBF), cerebral blood volume (CBV) and mean transit time (MTT) were measured in the gray matter (GM), white matter (WM) and the subcortical GM (SGM). The severity of CCSVI was assessed according to the venous hemodynamic insufficiency severity score (VHISS) on the basis of the number of venous segments exhibiting flow abnormalities. There was a significant association between increased VHISS and decreased CBF in the majority of examined regions of the brain parenchyma in MS patients. The most robust correlations were observed for GM and WM (r = -0.70 to -0.71, P < 0.002 and P corrected = 0.022), and for the putamen, thalamus, pulvinar nucleus of thalamus, globus pallidus and hippocampus (r = -0.59 to -0.71, P < 0.01 and P corrected < 0.05). No results for correlation between VHISS and CBV or MTT survived multiple comparison correction. This pilot study is the first to report a significant relationship between the severity of CCSVI and hypoperfusion in the brain parenchyma. These preliminary findings should be confirmed in a larger cohort of MS patients to ensure that they generalize to the MS population as a whole. Reduced perfusion could contribute to the known mechanisms of virtual hypoxia in degenerated axons.


Management of a departmental budget using an electronic spreadsheet.
Many dental school departments utilize computers, which provide the chairperson with important information on the status of the departmental budget. With the appropriate software, awareness of the funds remaining in each of the budget categories allows rapid decisions on a variety of issues. A computer-driven spreadsheet can be indispensable, providing essential information on current income and disbursements, and facilitating comparison with previous, current, and projected operations.


Application of engineered myoglobins for biosynthesis of clofazimine by integration with chemical synthesis.
Significant efforts have been made in the design of artificial metalloenzymes. Myoglobin (Mb), an O2 carrier, has been engineered to exhibit different functions. Herein, we applied a series of engineered Mb mutants with peroxidase activity for biosynthesis of clofazimine (CFZ), a potential drug with a broad-spectrum antiviral activity, by integration with chemical synthesis. Two of those mutants, F43Y Mb and F43Y/T67R Mb, have been shown to efficiently catalyze the oxidative coupling of 2-N-(4-chlorophenyl) benzene-1,2-diamine (N-4-CPBDA) in the presence of H2O2, with 97% yields. The overall catalytic efficiency (kcat/Km) is 46-fold and 82-fold higher than that of WT Mb, respectively. By further combination of this reaction with chemical synthesis, the production of CFZ was accomplished with an isolated yield of 72%. These results showed that engineered Mbs containing the Tyr-heme cross-link (F43Y Mb and F43Y/T67R Mb) exhibit enhanced activity in the oxidative coupling reaction. This study also indicates that the combination of biocatalysis and chemical synthesis avoids the need for the separation of intermediate products, which offers a convenient approach for the total synthesis of the biological compound CFZ.


Is DVT prophylaxis overemphasized? A randomized prospective study.
This study was designed to prospectively evaluate a previously published prognostic index for predicting deep venous thrombosis (DVT) in general surgical patients with conventional prophylaxis. Patients undergoing procedures of at least 1 hr duration (abdominal, thoracic, head and neck, inguinal) requiring general or spinal anesthetic were prospectively randomized into the following groups: Group 1, sequential pneumatic compression devices during surgery and 2 days postoperatively; Group 2, subcutaneous heparin (5000 U q 12 hr) starting 1 hr before surgery and for 7 days postop; Group 3, control group. All patients underwent duplex evaluation of bilateral lower extremity deep venous systems preoperatively and on postoperative Days 1, 3, and 30. In addition, a previously developed predictive DVT incidence indicator, the prognostic index (PI), was calculated for each patient. A total of 137 patients were entered into the study with 29 removed for patient/staff reasons. There were no differences in PI among the three groups at the 0.05 level (ANOVA). The distribution of risk factors for DVT including increased age, body size, hemoglobin (Hb), and colorectal procedures were distributed evenly among the groups. Additional factors such as diabetes, COPD, PVD, immobilization, and cancer were also evenly distributed among the groups. The PI predicted a 20% incidence of DVT. For Groups 1 (n = 25), 2 (n = 38), and 3 (n = 45) no DVTs were detected over the 30 days of study. During the study period, 8 DVTs were detected by duplex evaluation in general surgical patients not in the study (1.5%). In conclusion, in a prospective randomized study using sequential pneumatic compression devices, subcutaneous heparin or no prophylaxis in matched general surgical patients at moderate to high risk for thromboembolism, no DVTs occurred for up to 30 days. Furthermore, neither a PI nor other factors associated with DVT accurately predicted the incidence of DVT in this patient population.


Helium-3 from the mantle: primordial signal or cosmic dust?
Helium-3 in hotspot magmas has been used as unambiguous evidence for the existence of a primordial, undegassed reservoir deep in the Earth's mantle. However, a large amount of helium-3 is delivered to the Earth's surface by interplanetary dust particles (IDPs). Recycling of deep-sea sediments containing these particles to the mantle, and eventual incorporation in magma, can explain the high helium-3/helium-4 ratios of hotspot magmas. Basafts with high helium-3/helium-4 ratios may represent degassing of helium introduced by ancient (probably 1.5 to 2.0 billion years old) pelagic sediments rather than degassing of primordial lower mantle material brought to the surface in plumes. Influx of IDPs can also explain the neon and siderophile compositions of mantle samples.


The Jak-STAT pathway stimulated by interferon gamma.
Different cytokines stimulate the Janus kinase (Jak) and signal transducer and activator of transcription (STAT) pathway. Interferon gamma (IFN-gamma) is a type II interferon that triggers antiviral and adaptive immune responses through a Jak-STAT signaling pathway. These pathways involve a similar paradigm whereby cytokine binding to the receptor activates the Jaks, which phosphorylate STATs leading to their dimerization, translocation to the nucleus, and activation of their transcription factor activity. The animation shows binding of IFN-gamma and dimerization of the receptors, activation of Jak1 and Jak2, and phosphorylation of STAT1 leading to dimerization, translocation to the nucleus, and stimulation of target gene transcription.


Performance measurement for schizophrenia: adherence to guidelines for antipsychotic dose.
This report describes the development, application, and exploratory evaluation of a clinical performance measure based on recently published schizophrenia guidelines for antipsychotic dose. The performance measure, which assesses adherence to antipsychotic dose recommendations for acute schizophrenia treatment, was calculated at hospital discharge for 116 patients with schizophrenia who had participated in a 6-month outcomes study. The Brief Psychiatric Rating Scale (BPRS) was used to assess symptom severity at 6-month followup. At discharge, almost one-half of the patients were prescribed doses outside the recommended range. For the entire sample, linear regression models showed that the performance measure variable was not significantly associated with followup symptom severity (BPRS total scores). However, a significant association was observed for patients prescribed oral antipsychotics only (n = 69). Patients prescribed recommended doses had lower adjusted mean BPRS totals than patients prescribed doses either greater than (P < 0.05) or less than (P < 0.05) recommended. Our findings suggest that the antipsychotic dose performance measure may be useful for monitoring quality. It assesses a modifiable aspect of care for which clinical improvement is needed, and such improvement is likely to improve patient outcomes. Future research is needed to confirm our findings and to develop and test interventions to improve the quality of care for schizophrenia that incorporate this clinical performance measure.


Wingless/int-1induced secreted protein-1: a new biomarker for renal fibrosis.
This study aimed to detect the expression of Wnt-induced secreted protein-1 (WISP-1) in renal fibrosis (RF) and to clarify the underlying mechanism. An in vivo mousee model of unilateral ureteral obstruction (UUO) and in vitro model of fibrosis on renal tubular epithelial NRK52E cells after transforming growth factor-β1 (TGF-β1) stimulation were used. Quantitative real-time PCR (qRT-PCR), Western blot (WB), and immunohistochemistry were used to detect WISP-1 and fibrosis markers, including the expression of fibronectin (FN), collagen I (Col I), collagen IV (Col IV), and α-smooth muscle actin (α-SMA). In vitro experiments showed that the expression of WISP-1 and fibrosis markers FN, Col I, Col IV, and α-SMA in rat renal tubular epithelial cells were significantly higher than that in the control group after 48 h of TGF-β1 stimulation. In vivo experiments showed that the expressions of WISP-1 and fibrosis markers FN, Col I, Col IV, and α-SMA in the obstructed kidney of UUO animal models were significantly increased in mRNA and protein levels compared to normal mice. This study showed that WISP-1 may be an essential cytokine that promotes renal fibrosis, being involved in the development of renal fibrosis.


Childhood Parental Incarceration and Adult-Onset Hypertension and Cardiovascular Risk.
Parental incarceration is an adverse childhood experience that disproportionately affects racially minoritized individuals and has been associated with long-term health risks. Although cardiovascular disease remains the primary cause of mortality differences between Black and White individuals in the US, the association between parental incarceration and cardiovascular risk remains poorly understood. To examine the association between parental incarceration during childhood and incident cardiovascular risk in adulthood. This population-based cohort study included data from waves IV (2008-2009) and V (2016-2018) of the US National Longitudinal Study of Adolescent to Adult Health. Participants represented US adults transitioning from young adulthood to adulthood. Data were analyzed from October 28, 2021, to May 1, 2023. Parental incarceration was defined as a parent or parent-like figure going to jail or prison when participants were aged younger than 18 years. Outcome measures included self-reported diagnoses of obesity, hyperlipidemia, hypertension, diabetes, or heart disease as well as serum elevations in non-high-density lipoprotein cholesterol (≥160 mg/dL) and high-sensitivity C-reactive protein (hsCRP >3 mg/L), a marker of inflammation used to estimate risk of future coronary events. Using sampling weights, incident development of each outcome was modeled as a function of parental incarceration, adjusting for participant- and neighborhood-level characteristics. This study included 9629 participants representing 16 077 108 US adults. Approximately half of participants were women (5498 [weighted 50.3%]) and the majority (5895 [weighted 71.4%]) were White. The mean participant age was 37.8 years (95% CI, 37.5 to 38.0 years) in wave V compared with 28.9 years (95% CI, 28.6 to 29.1 years) in wave IV. In wave V, those with childhood exposure to parental incarceration had lower educational attainment (91 [weighted 8.2%] vs 245 [weighted 4.2%] completing less than high school), had higher rates of public insurance (257 [weighted 20.6%] vs 806 [weighted 11.0%]), and were disproportionately Black (374 [weighted 22.5%] vs 1488 [weighted 13.6%]). Parental incarceration was associated with 33% higher adjusted odds (95% CI, 1.05 to 1.68) of developing hypertension and 60% higher adjusted odds (95% CI, 1.03 to 2.48) of developing elevated hsCRP. Associations between childhood parental incarceration and other diagnoses (ie, obesity, hyperlipidemia, diabetes, or heart disease) and serum lipid levels were not observed. In this cohort study of US adults transitioning from young adulthood to adulthood, an increased incidence of hypertension and high-risk hsCRP, but not other cardiovascular risk factors, was observed among those exposed to parental incarceration during childhood. These findings suggest possible transgenerational health consequences of mass incarceration.


A call for a moratorium on the .health generic top-level domain: preventing the commercialization and exclusive control of online health information.
In just a few weeks, the Internet could be expanded to include a new .health generic top-level domain name run by a for-profit company with virtually no public health credentials - unless the international community intervenes immediately. This matters to the future of global public health as the "Health Internet" has begun to emerge as the predominant source of health information for consumers and patients. Despite this increasing use and reliance on online health information that may have inadequate quality or reliability, the Internet Corporation for Assigned Names and Numbers (ICANN) recently announced it intends to move forward with an auction to award the exclusive, 10 year rights to the .health generic top-level domain name. This decision is being made over the protests of the World Medical Association, World Health Organization, and other stakeholders, who have called for a suspension or delay until key questions can be resolved. However, rather than engage in constructive dialogue with the public health community over its concerns, ICANN chose the International Chamber of Commerce-a business lobbying group for industries to adjudicate the .health concerns. This has resulted in a rejection of challenges filed by ICANN's own independent watchdog and others, such that ICANN's Board decided in June 2014 that there are "no noted objections to move forward" in auctioning the .health generic top-level domain name to the highest bidder before the end of the year. This follows ICANN's award of several other health-related generic top-level domain names that have been unsuccessfully contested. In response, we call for an immediate moratorium/suspension of the ICANN award/auction process in order to provide the international public health community time to ensure the proper management and governance of health information online.


Osteopontin promotes the progression of gastric cancer through the NF-κB pathway regulated by the MAPK and PI3K.
To elucidate the biological functions of osteopontin (OPN) in gastric tumors and to better understand the molecular events of OPN responsible for the malignancy, the present studies were performed. Growth curve, apoptosis assay, invasion assay and migration assay revealed that OPN status significantly affected proliferation, apoptosis, invasion and migration in gastric cancer cell lines. In mouse xenograft models of human gastric cancer, OPN silencing significantly inhibited tumor growth and the incidence of metastasis compared with non-silenced control. Mechanistic investigations revealed that OPN silencing inhibited the mitogen-activated protein kinase (MAPK), phosphoinositide 3-kinase (PI3K) and phosphoinositide 3-kinase (NF-κB) pathways, and OPN-mediated NF-κB activity was reduced in the presence of MAPK or PI3K inhibitor. Our findings also indicated that OPN through the NF-κB pathway promotes the expression of matrix metalloproteinase 2 (MMP-2), MMP-9 and urokinase-type plasminogen activator (uPA), the activation of MMP-2 and MMP-9, and the inhibition of caspase-3. Taken together, our results reveal that OPN promotes the progression of gastric cancer through the NF-κB pathway, which is regulated by the MAPK and PI3K pathways and leads to MMP-2, MMP-9 and uPA activation and caspase-3 inhibition. These findings identify OPN as a novel oncogene in gastric cancer and suggest that OPN is an attractive therapeutic target for this aggressive malignancy.


Method validation and proficiency testing for determination of total arsenic in apple juice by inductively coupled plasma/ mass spectrometry.
A proficiency test (PT) program for determination of total As in apple juice samples was conducted by the Food Emergency Response Network (FERN) laboratories. An analytical method using inductively coupled plasma (ICP)-MS was validated for this project. The LOD and LOQ were determined to be 0.315 and 2.32 ng/g, respectively. A total of eight apple juice samples were sent to 38 FERN laboratories, and results were statistically evaluated according to ISO 13528:2005. The total As concentrations in the PT samples reported by the participating laboratories were very close to those obtained in the homogeneity and stability tests. The reproducibility, repeatability, interlaboratory, and intralaboratory variability results led to 69% of participating laboratories being rated as satisfactory using the widely accepted Izl score <or= 2 criterion. The percentage of laboratories with acceptable z scores (Izl scores <or=2) increased from 61 to 89% with the increase in As concentration in PT samples from 5.05 to 30.72 ng/g. This study demonstrated that the ICP/MS method was adequate for total As determination in fruit juice, and most of the participating laboratories were proficient in their analytical performance.


Two overlapping domains of a lyssavirus matrix protein that acts on different cell death pathways.
The lyssavirus matrix (M) protein induces apoptosis. The regions of the M protein that are essential for triggering cell death pathways are not yet clearly defined. We therefore compared the M proteins from two viruses that have contrasting characteristics in terms of cellular apoptosis: a genotype 3 lyssavirus, Mokola virus (MOK), and a genotype 1 rabies virus isolated from a dog from Thailand (THA). We identified a 20-amino-acid fragment (corresponding to positions 67 to 86) that retained the cell death activities of the full-length M protein from MOK via both the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and inhibition of cytochrome c oxidase (CcO) activity. We found that the amino acids at positions 77 and 81 have an essential role in triggering these two cell death pathways. Directed mutagenesis demonstrated that the amino acid at position 77 affects CcO activity, whereas the amino acid at position 81 affects TRAIL-dependent apoptosis. Mutations in the full-length M protein that compromised induction of either of these two pathways resulted in delayed apoptosis compared with the time to apoptosis for the nonmutated control.


Danish cattle veterinarians' perspectives on antimicrobial use: Contextual and individual influencing factors.
The global risk for antimicrobial resistance (AMR) can be reduced by reducing antimicrobial use (AMU). Veterinarians are one of the key actors in relation to AMU in livestock, and understanding the dynamics of veterinary treatment and prescription is central to achieving AMU reduction. Veterinary AMU decisions are influenced by a complex pattern of both individual and contextual factors. In this study, we conducted semi-structured interviews with 16 Danish cattle veterinarians to investigate their perspectives on AMU and current practices in a national context with low use and extensive legal control. We found that personal experiences and emotions, rather than scientific evidence, guided some veterinarians when making AMU choices. Furthermore, less-experienced veterinarians felt pressure to prescribe according to colleagues' and farmers' preferences for certain antimicrobials. We found that changes in Danish legislation seemed to have introduced hesitancy and a lack of motivation within the veterinary profession, and that AMR was perceived as an abstract threat not applicable to the veterinarians' daily professional decision making. We concluded that the lack of field-generated research of local relevance nourished a culture in which AMU choices are built on personal experience rather than scientific evidence, which also diminished newly educated veterinarians' self-confidence in relation to their AMU choices. Future research should focus on developing locally relevant research on optimal AMU choices and AMR, and the implications of extensive legal control of AMU in livestock farming should be further investigated to find a balance on the path to reducing AMU.


Low-pathogenicity avian influenza virus in live bird markets--what about the livestock area?
Low-pathogenic avian influenza virus (AV) continues to be isolated from the live bird markets (LBMs) in the Northeasten United States. Recent years have seen increasing numbers of these markets opening and an expansion of the type of animals they sell in conjunction with traditional live poultry. Specific-pathogen-free chickens were released into the livestock area of 13 New York City LBMs and then tested for evidence of AIV. We were able to recover virus or demonstrate seroconversion among the chickens introduced to four of the markets.


The distribution of nuclear mass, volume and dimensions.
Our own measurements and a review of measurements presented in the literature showed that the mass, volume and dimensions of cell nuclei are distributed with a skew towards higher values, a distribution that can best be described as lognormal. The practical consequences of this finding suggest that the measurements of mass and of size should be plotted on a logarithmic scale while DNA values are appropriately presented on a linear scale. The distribution of DNA values of normal cells represents errors introduced by random disturbances in preparations and measurements.


Trust and risk communication in high-risk organizations: a test of principles from social risk research.
This study explored the effects of open communication about occupational risks on workers' trust beliefs and trust intentions toward risk management, and the resilience of these beliefs and intentions to further risk information. An experimental survey of 393 student nurses showed the importance of open communication in the development of worker trust in risk management. Consistent with the trust asymmetry principle, we found that the increase in trust beliefs following open communication was weaker than the reduction in trust following a lack of communication. Further, the level of trust developed through communication (or lack of) influenced the way that subsequent risk information was processed. Negative risk information reduced trust beliefs in nurses with already low levels of trust while positive risk information increased trust beliefs only in those with already high levels. A similar pattern of results emerged for nurses' trust intentions, although the magnitude of these effects was weaker. The implications of these findings for occupational risk management are discussed.


[Common sense in operative treatment of thoracoabdominal aortic aneurysms].
Treatment approaches of patients with thoracoabdominal aneurysms and aortic dissections type B should be individual. Risk ratio of surgery itself and progression of non-operated disease, such as aneurysm rupture, is the determining factor in defining pro et contra surgical treatment.


Role of osteoclast extracellular signal-regulated kinase (ERK) in cell survival and maintenance of cell polarity.
Morphological changes of osteoclasts by a MEK1 inhibitor, PD98059, were investigated to clarify a role of ERK. PD98059 promoted apoptosis of osteoclasts and the loss of ruffled borders. This study supports the importance of ERK in survival and polarity of osteoclasts. Extracellular signal-regulated kinase (ERK) is a mitogen activated protein kinase (MAPK) that has been reported to play a role in the survival and apoptosis of osteoclasts. However, the precise signal transduction mechanism is not fully understood. The aim of this study was to clarify the role of ERK in osteoclasts by histological analysis. Using a rat calvarial organ culture system, the inhibition of ERK phosphorylation by PD98059, a MAPK/ERK kinase 1 (MEK1) inhibitor, was assayed by immunoblotting. Morphological changes in osteoclasts induced by PD98059 were elucidated by light and electron microscopy. The cellular localization of ERK was also determined by immunoelectron microscopy. PD98059 inhibited phosphorylated ERK after a 1-h incubation. Ultrastructural study demonstrated that PD98059 induced the accumulation of vesicles and vacuoles in osteoclasts and the loss of ruffled border at 1 h. At 3 h, some osteoclasts showed apoptosis with nuclear condensation, and at 6 h after PD98059 treatment, many osteoclasts were detached from the bone surface and had lost their cell polarity. Electron microscopic immunohistochemistry revealed that ERK was mainly localized in the cytoplasm of clear zones in control osteoclasts, but apoptotic osteoclasts also showed immunoreactivity in clear zone-like structures in contact with osteoblast-lineage cells. These findings indicate that ERK in osteoclasts is involved in their survival and may be involved in the formation of a ruffled border and the maintenance of cell polarity.


Graft vascular function after transplantation of pancreatic islets.
Endogenous pancreatic islets have a dense glomerular-like angioarchitecture, which ensures an optimal delivery of oxygen and nutrients to the islet cells, provides signals from other cells in the body and disposes secreted hormones. Transplantation of isolated islets means that their vascular connection is interrupted. The islet grafts therefore depend upon endothelial cells and microvessels originating in the implantation organ for derivation of a new vascular system. A re-establishment of islet blood-flow occurs within 7-14 days after transplantation, mainly through vascular sprouting. The newly formed blood vessels acquire the morphological characteristics of those in endogenous islets. In intraportally transplanted islets to the liver, the islets become revascularized almost exclusively from tributaries to the hepatic artery. Exocrine contamination of the transplanted islets could hamper the revascularization process, whereas neither cryopreservation nor immunosuppressive drugs like cyclosporin, prednisolon and RS-61443 have any essential effects on the angiogenesis. Investigators have noticed improvements in islet graft survival and function by means of basic fibroblast growth factor (bFGF), acidic FGF and endothelial cell growth factor exposure of the grafts. The functional properties of transplanted islets are largely unknown, but evidence from experimental islet transplantation suggests that both the blood perfusion and the tissue oxygen tension of the grafted islets are chronically decreased, indicating an insufficient vascular system. In order to achieve optimal condition for survival and function of transplanted beta cells, it is important to ascertain whether impairments in vascular function are present also after clinical islet transplantations as well.


Probabilistic algorithms for MEG/EEG source reconstruction using temporal basis functions learned from data.
We present two related probabilistic methods for neural source reconstruction from MEG/EEG data that reduce effects of interference, noise, and correlated sources. Both methods localize source activity using a linear mixture of temporal basis functions (TBFs) learned from the data. In contrast to existing methods that use predetermined TBFs, we compute TBFs from data using a graphical factor analysis based model [Nagarajan, S.S., Attias, H.T., Hild, K.E., Sekihara, K., 2007a. A probabilistic algorithm for robust interference suppression in bioelectromagnetic sensor data. Stat Med 26, 3886-3910], which separates evoked or event-related source activity from ongoing spontaneous background brain activity. Both algorithms compute an optimal weighting of these TBFs at each voxel to provide a spatiotemporal map of activity across the brain and a source image map from the likelihood of a dipole source at each voxel. We explicitly model, with two different robust parameterizations, the contribution from signals outside a voxel of interest. The two models differ in a trade-off of computational speed versus accuracy of learning the unknown interference contributions. Performance in simulations and real data, both with large noise and interference and/or correlated sources, demonstrates significant improvement over existing source localization methods.


Trehalose accumulation during cellular stress protects cells and cellular proteins from damage by oxygen radicals.
The disaccharide trehalose, which accumulates dramatically during heat shock and stationary phase in many organisms, enhances thermotolerance and reduces aggregation of denatured proteins. Here we report a new role for trehalose in protecting cells against oxygen radicals. Exposure of Saccharomyces cerevisiae to a mild heat shock (38 degrees C) or to a proteasome inhibitor (MG132) induced trehalose accumulation and markedly increased the viability of the cells upon exposure to a free radical-generating system (H(2)O(2)/iron). When cells were returned to normal growth temperature (28 degrees C) or MG132 was removed from the medium, the trehalose content and resistance to oxygen radicals decreased rapidly. Furthermore, a mutant unable to synthesize trehalose was much more sensitive to killing by oxygen radicals than wild-type cells. Providing trehalose exogenously enhanced the resistance of mutant cells to H(2)O(2). Exposure of cells to H(2)O(2) caused oxidative damage to amino acids in cellular proteins, and trehalose accumulation was found to reduce such damage. After even brief exposure to H(2)O(2), the trehalose-deficient mutant exhibited a much higher content of oxidatively damaged proteins than wild-type cells. Trehalose accumulation decreased the initial appearance of damaged proteins, presumably by acting as a free radical scavenger. Therefore, trehalose accumulation in stressed cells plays a major role in protecting cellular constituents from oxidative damage.


Outcomes associated with supportive periodontal therapy in smokers and nonsmokers.
This study evaluated the effects of smoking status on retrospective clinical and radiographic measures of periodontal disease and compared these to prospective changes in digital radiographic bone height. Clinical data on moderate (4 to 6 mm) and severe (> 6 mm) periodontal pocket depths, and bleeding on probing, were obtained from 95 subjects on suggested three-month supportive periodontal therapy (SPT) for AAP Class III/IV periodontitis. Standardized radiographic data were obtained concerning posterior interproximal alveolar bone height from 36 of the 95 subjects using computer-assisted digital technology at baseline and one year later. The subjects were divided into groups by smoking status: current, former, and never. Data were evaluated using a general linear statistical model. Evaluation of clinical data showed that current smokers exhibited a significantly higher percentage of moderate (18%) and severe (1%) periodontal pockets than nonsmokers (10% and 0%, respectively; p < 0.002). Baseline radiographic interproximal bone height loss also was greater in current smokers (5.75 +/- 1.07 v. 4.64 +/- 1.16 mm). Bone loss over one year occurred in 5% of the sites, but was not significantly different among groups. Clinical periodontal pockets and bone loss accumulated more rapidly in smokers, even though they submitted to regular supportive periodontal therapy. Although this population was clinically compliant over a one year period, digital radiography showed a high incidence of detectable bone loss. The impact of smoking, however, may require longer than one year to show longitudinal changes. It is recommended that a periodic radiographic analysis on bone height be considered during SPT, and longer term studies be conducted in order to accurately identify the outcome of smoking status on this variable.


Composition, Mineral and Fatty Acid Profiles of Milk from Goats Fed with Different Proportions of Broccoli and Artichoke Plant By-Products.
In the Mediterranean region, artichoke and broccoli are major crops with a high amount of by-products that can be used as alternative feedstuffs for ruminants, lowering feed costs and enhancing milk sustainability while reducing the environmental impact of dairy production. However, nutritional quality of milk needs to be assured under these production conditions and an optimal inclusion ratio of silages should be determined. This work aimed to evaluate the effect of three inclusion levels (25%, 40%, and 60%) of these silages (artichoke plant, AP, and broccoli by-product, BB) in goat diets on milk yield, composition, and mineral and fatty profiles. Treatments with 60% inclusion of AP and BB presented the lowest milk yield. No differences were found on the milk mineral profile. Inclusion of AP in the animals' diet improved the milk lipid profile from the point of view of human health (AI, TI) compared to BB due to a lower saturated fatty acid content (C12:0, C14:0, and C16:0) and a higher concentration of polyunsaturated fatty acids (PUFA), especially vaccenic acid (C18:1 trans11) and rumenic acid (CLA cis9, trans11), without any differences with the control treatment.


Fellowship training in extracorporeal life support: Characterization and educational needs assessment.
Adult Extracorporeal Life Support (ECLS) use is rapidly increasing. The structure of fellowship ECLS education is unknown. We sought to define current ECLS education and identify curricular needs. An anonymous survey with Likert, binary and free response questions was sent to Critical Care Program Directors (CCPDs). A total of 103 CCPDs responded, a response rate of 31. ECLS training was provided by 64% (66/103) of fellowships. Importantly, 50% (52/103) of CCPDs agreed or strongly agreed that fellows should be competent in ECLS and 70% (72/103) agreed or strongly agreed that ECLS will be an important part of critical care in the next 10years. Only 28% (29/103) and 37% (38/103) of CCPDs agreed or strongly agreed their fellows could independently manage veno-arterial or veno-venous ECLS, respectively. Formal ECLS education was 5h or less in 85% (88/103) of programs. Desired curricular improvements were: simulation 50% (51/103), patient volume 47% (48/103), and didactics 44% (45/103). CCPDs identified ECLS as a critical care skill, but believe that a minority of fellows are prepared for independent practice. Simulation, formal didactics and clinical volume are key needs. These data will guide the development of ECLS curriculum.


Clinical results of modular polyethylene insert exchange with retention of total knee arthroplasty components.
Modular polyethylene inserts have enabled surgeons to perform an isolated tibial insert exchange while retaining well fixed components. The purpose of this study was to review the results of insert revision and to clarify the role of this option compared with that of revision total knee arthroplasty. Fifty-six patients (sixty-three knees) were managed with revision of a tibial polyethylene insert and retention of well aligned and stable femoral and tibial components. The implants had been in situ for an average of fifty-nine months (range, two to 108 months) at the time of the insert exchange. The inserts that were removed at the time of exchange were evaluated with regard to wear of the articular surface according to the classification system of Hood et al. and with regard to undersurface wear according to the method described by Wasielewski et al. Forty-eight knees were followed for an average of 7.4 years (range, 3.0 to 12.2 years) after the insert exchange. Knees that did not require an additional operation were considered to have had a successful exchange. Seven of the forty-eight exchanges failed, at an average of fifty-four months, because of accelerated wear of the new insert. All seven knees required complete revision of all components. Of the twenty-two exchanges that were performed because of severe wear of the primary insert, six (27 percent) failed at an average of less than five years; thus, knees in which the exchange was performed because of advanced wear were more likely to fail again (p < 0.05). In addition, primary inserts that were removed from knees in which the exchange procedure subsequently failed had higher delamination scores than those that were removed from knees in which the exchange was successful (p < 0.05). Most of the primary inserts had substantial undersurface wear at the time of the exchange procedure. Metallosis (thirty knees) and osteolysis (nineteen knees) were unrelated to failure of the exchange. An isolated revision of the tibial polyethylene insert should not be performed when there is accelerated wear of the insert with severe delamination and grade-3 or 4 undersurface wear within ten years after the primary procedure. Because a variety of patient-related, implant-related, and technical factors influence polyethylene wear, the orthopaedist must consider multiple variables whenever contemplating a limited revision.


Portals to Wonderland: health portals lead to confusing information about the effects of health care.
The Internet offers a seemingly endless amount of health information of varying quality. Health portals, which provide entry points to quality-controlled collections of websites, have been hailed as a solution to this problem. The objective of this study is to assess the extent to which government-run health portals provide access to relevant, valid and understandable information about the effects of health care. We selected eight clinically relevant questions for which there was a systematic review, searched four portals for answers, and compared the answers we found to the results of the systematic reviews. Our searches resulted in 3400 hits, 155 of which mentioned both the condition and the intervention in one of the eight questions. Sixty-three of the 155 web pages did not give any information about the effect of the intervention. Seventy-seven qualitatively described the effects of the intervention. Twenty-six of these had information that was too unclear to be categorised; 15 were not consistent with the systematic review; and 36 were consistent with the review, but usually did not mention what happens without the intervention, what outcomes have been measured or when they were measured. Fifteen web pages quantitatively described effects. Four of these were abstracts from the systematic review, nine had information that was incomplete and potentially misleading because of a lack of information about people not receiving the intervention and the length of follow-up; one had information that was consistent with the review, but only referred to three trials whereas the review included six; and one was consistent with the review. Information accessible through health portals is unlikely to be based on systematic reviews and is often unclear, incomplete and misleading. Portals are only as good as the websites they lead to. Investments in national health portals are unlikely to benefit consumers without investments in the production and maintenance of relevant, valid and understandable information to which the portals lead.


Maternal Immune Activation Alters Fetal Brain Development and Enhances Proliferation of Neural Precursor Cells in Rats.
Maternal immune activation (MIA) caused by exposure to pathogens or inflammation during critical periods of neurodevelopment is a major risk factor for behavioral deficits and psychiatric illness in offspring. A spectrum of behavioral abnormalities can be recapitulated in rodents by inducing MIA using the viral mimetic, PolyI:C. Many studies have focused on long-term changes in brain structure and behavioral outcomes in offspring following maternal PolyI:C exposure, but acute changes in prenatal development are not well-characterized. Using RNA-Sequencing, we profiled acute transcriptomic changes in rat conceptuses (decidua along with nascent embryo and placenta) after maternal PolyI:C exposure during early gestation, which enabled us to capture gene expression changes provoked by MIA inclusive to the embryonic milieu. We identified a robust increase in expression of genes related to antiviral inflammation following maternal PolyI:C exposure, and a corresponding decrease in transcripts associated with nervous system development. At mid-gestation, regions of the developing cortex were thicker in fetuses prenatally challenged with PolyI:C, with females displaying a thicker ventricular zone and males a thicker cortical mantle. Along these lines, neural precursor cells (NPCs) isolated from fetal brains prenatally challenged with PolyI:C exhibited a higher rate of self-renewal. Expression of Notch1 and the Notch ligand, delta-like ligand 1, which are both highly implicated in maintenance of NPCs and nervous system development, was increased following PolyI:C exposure. These results suggest that MIA elicits rapid gene expression changes within the conceptus, including repression of neurodevelopmental pathways, resulting in profound alterations in fetal brain development.


The GTPase Rab26 links synaptic vesicles to the autophagy pathway.
Small GTPases of the Rab family not only regulate target recognition in membrane traffic but also control other cellular functions such as cytoskeletal transport and autophagy. Here we show that Rab26 is specifically associated with clusters of synaptic vesicles in neurites. Overexpression of active but not of GDP-preferring Rab26 enhances vesicle clustering, which is particularly conspicuous for the EGFP-tagged variant, resulting in a massive accumulation of synaptic vesicles in neuronal somata without altering the distribution of other organelles. Both endogenous and induced clusters co-localize with autophagy-related proteins such as Atg16L1, LC3B and Rab33B but not with other organelles. Furthermore, Atg16L1 appears to be a direct effector of Rab26 and binds Rab26 in its GTP-bound form, albeit only with low affinity. We propose that Rab26 selectively directs synaptic and secretory vesicles into preautophagosomal structures, suggesting the presence of a novel pathway for degradation of synaptic vesicles.


Buprenorphine decreases the CCL2-mediated chemotactic response of monocytes.
Despite successful combined antiretroviral therapy, ∼ 60% of HIV-infected people exhibit HIV-associated neurocognitive disorders (HAND). CCL2 is elevated in the CNS of infected people with HAND and mediates monocyte influx into the CNS, which is critical in neuroAIDS. Many HIV-infected opiate abusers have increased neuroinflammation that may augment HAND. Buprenorphine is used to treat opiate addiction. However, there are few studies that examine its impact on HIV neuropathogenesis. We show that buprenorphine reduces the chemotactic phenotype of monocytes. Buprenorphine decreases the formation of membrane projections in response to CCL2. It also decreases CCL2-induced chemotaxis and mediates a delay in reinsertion of the CCL2 receptor, CCR2, into the cell membrane after CCL2-mediated receptor internalization, suggesting a mechanism of action of buprenorphine. Signaling pathways in CCL2-induced migration include increased phosphorylation of p38 MAPK and of the junctional protein JAM-A. We show that buprenorphine decreases these phosphorylations in CCL2-treated monocytes. Using DAMGO, CTAP, and Nor-BNI, we demonstrate that the effect of buprenorphine on CCL2 signaling is opioid receptor mediated. To identify additional potential mechanisms by which buprenorphine inhibits CCL2-induced monocyte migration, we performed proteomic analyses to characterize additional proteins in monocytes whose phosphorylation after CCL2 treatment was inhibited by buprenorphine. Leukosialin and S100A9 were identified and had not been shown previously to be involved in monocyte migration. We propose that buprenorphine limits CCL2-mediated monocyte transmigration into the CNS, thereby reducing neuroinflammation characteristic of HAND. Our findings underscore the use of buprenorphine as a therapeutic for neuroinflammation as well as for addiction.


In vitro analysis of transport and metabolism of 4'-thiothymidine in human tumor cells.
The use of thymidine (TdR) and thymidine analogs such as 3'-fluoro-3'-deoxythymidine (FLT) as positron emission tomography (PET)-based proliferation markers can provide information on tumor response to treatment. Studies on another TdR analog, 4'-thiothymidine (4DST), suggest that it might be a better PET-based proliferation tracer than either TdR or FLT. 4DST is resistant to the catabolism that complicates analysis of TdR in PET studies, but unlike FLT, 4DST is incorporated into DNA. To further evaluate 4DST, the kinetics of 4DST transport and metabolism were determined and compared to FLT and TdR. Transport and metabolism of FLT, TdR and 4DST were examined in the human adenocarcinoma cell line A549 under exponential-growth conditions. Single cell suspensions were incubated in buffer supplemented with radiolabeled tracer in the presence or absence of nitrobenzylmercaptopurine ribonucleoside (NBMPR), an inhibitor of equilibrative nucleoside transporters (ENT). Kinetics of tracer uptake was determined in whole cells and tracer metabolism measured by high performance liquid chromatography of cell lysates. TdR and 4DST were qualitatively similar in terms of ENT-dependent transport, shapes of uptake curves, and relative levels of DNA incorporation. FLT did not incorporate into DNA, showed a significant temperature effect for uptake, and its transport had a significant NBMPR-resistant component. Overall 4DST metabolism was significantly slower than either TdR or FLT. 4DST provides a good alternative for TdR in PET and has advantages over FLT in proliferation measurement. However, slow 4DST metabolism and the short half-life of the (11)C label might limit widespread use in PET.


Cyclobutane pyrimidine dimers and (6-4) photoproducts block polymerization by DNA polymerase I.
Bipyrimidine cyclobutane dimers and 6-4'-(pyrimidin-2'-one)-pyrimidine photoproducts are the major adducts formed in DNA following exposure to ultraviolet light. The relationship between the type and frequency of UV-induced DNA damage and the effects of such damage on DNA replication were investigated. UV-irradiated M13 phage DNA was employed in polymerization reactions with the Kenow fragment of Escherichia coli DNA polymerase I. The locations and frequencies of polymerase termination events occurring within a defined sequence of M13 DNA were compared with measurements of the locations and frequencies of UV-induced DNA damage of the same DNA sequence by using UV-specific enzymatic and chemical methods. The results indicate that both cyclobutane dimers and (6-4) photoproducts quantitatively block polymerization by DNA polymerase I.


Expression of thyroglobulin by regulatory T cells in thyroid tissue.
The follicles are the minimal functional unit of the thyroid; the morphology and the function of each follicle can vary significantly. However, the reasons for the apparent follicular heterogeneity are poorly understood. Some tissue-resident regulatory T cells (Tregs) have a special phenotype that expresses unique molecules related to local tissue and regulates the tissue functions. The aim of this study was to identify the phenotype of thyroid Tregs and the roles of thyroid Tregs in thyroid physiological regulation. Thyroid tissue and peripheral blood samples were obtained from patients with benign thyroid nodules. Microarray-based gene expression, flow cytometry, immunofluorescence microscopy, and functional analysis of thyroid Tregs were performed. Here, we demonstrated that human thyroid Tregs expressed high level of thyroglobulin (Tg), both gene and protein. The immunofluorescence microscopy of thyroid section showed that the FOXP3+Tg+ cells concentrated in some of the thyroid follicles, at the side of the thyroid follicle. The peripheral blood Tregs expressed minimal levels of Tg, and low levels of Tg could effectively induce peripheral blood Tregs to express Tg, which was independent of thyrotropin simulation. Furthermore, the Tg secreted freely from thyroid Tregs that negatively regulated some thyroid-related genes expression. Our results revealed that the thyroid Tregs was a distinct population of Tregs, which expressed high level of Tg. The thyroid Tregs regulate thyroid function by Tg that is paracrine from the cells.


The STIM1/Orai signaling machinery.
Ca(2+) influx via store-operated Ca(2+) release activated Ca(2+) (CRAC) channels represents a main signaling pathway for T-cell activation as well as mast-cell degranulation. The ER-located Ca(2+)-sensor, STIM1 and the Ca(2+)-selective ion pore, Orai1 in the membrane are sufficient to fully reconstitute CRAC currents. Their identification, but even more the recent structural resolution of both proteins by X-ray crystallography has substantially advanced the understanding of the activation mechanism of CRAC channels. In this review, we provide a detailed description of the STIM1/Orai1 signaling pathway thereby focusing on the critical domains mediating both, intra- as well as intermolecular interactions and on the ion permeation pathway. Based on the results of functional studies as well as the recently published crystal structures, we portray a mechanistic view of the steps in the CRAC channel signaling cascade ranging from STIM1 oligomerization over STIM1-Orai1 coupling to the ultimate Orai1 channel activation and permeation.


Roll-to-roll-compatible, flexible, transparent electrodes based on self-nanoembedded Cu nanowires using intense pulsed light irradiation.
Copper nanowire (Cu NW)-based flexible transparent conductive electrodes (FTCEs) have been investigated in detail for use in various applications such as flexible touch screens, organic photovoltaics and organic light-emitting diodes. In this study, hexadecylamine (HDA) adsorbed onto the surface of NWs is changed into polyvinylpyrrolidone (PVP) via a ligand exchange process; the high-molecular-weight PVP enables high dispersion stability. Intense pulsed light (IPL) irradiation is used to remove organic species present on the surface of the NWs and to form direct connections between the NWs rapidly without any atmospheric control. NWs are self-nanoembedded into a plastic substrate after IPL irradiation, which results in a smooth surface, strong NW/substrate adhesion, excellent mechanical flexibility and enhanced oxidation stability. Moreover, Cu NW FTCEs with high uniformities are successfully fabricated on a large area (150 mm × 200 mm) via successive IPL irradiation that is synchronized with the motion of the sample stage. This study demonstrates the possibility of roll-to-roll-based, large-scale production of low-cost, high-performance Cu NW-based FTCEs.


A gene expression signature identifies two prognostic subgroups of basal breast cancer.
Prognosis of basal breast cancers is poor but heterogeneous. Medullary breast cancers (MBC) display a basal profile, but a favorable prognosis. We hypothesized that a previously published 368-gene expression signature associated with MBC might serve to define a prognostic classifier in basal cancers. We collected public gene expression and histoclinical data of 2145 invasive early breast adenocarcinomas. We developed a Support Vector Machine (SVM) classifier based on this 368-gene list in a learning set, and tested its predictive performances in an independent validation set. Then, we assessed its prognostic value and that of six prognostic signatures for disease-free survival (DFS) in the remaining 2034 samples. The SVM model accurately classified all MBC samples in the learning and validation sets. A total of 466 cases were basal across other sets. The SVM classifier separated them into two subgroups, subgroup 1 (resembling MBC) and subgroup 2 (not resembling MBC). Subgroup 1 exhibited 71% 5-year DFS, whereas subgroup 2 exhibited 50% (P = 9.93E-05). The classifier outperformed the classical prognostic variables in multivariate analysis, conferring lesser risk for relapse in subgroup 1 (HR = 0.52, P = 3.9E-04). This prognostic value was specific to the basal subtype, in which none of the other prognostic signatures was informative. Ontology analysis revealed effective immune response (IR), enhanced tumor cell apoptosis, elevated levels of metastasis-inhibiting factors and low levels of metastasis-promoting factors in the good-prognosis subgroup, and a more developed cell migration system in the poor-prognosis subgroup. In conclusion, based on this 368-gene SVM model derived from an MBC signature, basal breast cancers were classified in two prognostic subgroups, suggesting that MBC and basal breast cancers share similar molecular alterations associated with aggressiveness. This signature could help define the prognosis, adapt the systemic treatment, and identify new therapeutic targets.


Similarity of secretory protein I from parathyroid gland to chromogranin A from adrenal medulla.
We have compared the amino acid and carbohydrate compositions, partial amino acid sequences, immunological crossreactivity, and physical properties of secretory protein I of the parathyroid gland and chromogranin A of adrenal gland. This comparison indicates that these proteins are similar molecules. Because secretory protein I is present in secretory granules containing parathormone and is cosecreted with the hormone, and because chromogranin A is contained within chromaffin granules and, likewise, is secreted with the catecholamines, the present observations raise the possibility that this class of protein plays a general role in hormone secretion or storage mechanisms.


Evaluation of Ischemic Heart Disease Factors in Hemophilia Patients in Khorramabad.
Evaluation of risk factors associated with coronary artery disease and cardiac health in hemophilia patients is necessary to prevent the onset of ischemic heart disease. In this study, we evaluated the cardiovascular status of hemophilic patients in Lorestan province for the early onset of ischemic heart disease. In this cross-sectional descriptive study, a total of 80 patients presenting severe hemophilia, a detailed questionnaire-based investigation was conducted to analyze the prevalence of cardiovascular risk factors in severe hemophilic patients. In patients with hemophilia, body mass index (BMI), blood pressure, diabetes, LDL, cholesterol and HDL, the risk of cardiovascular death was estimated using a predictive risk predictor algorithm of Europe SCORE. The mean age of the patients was 25 years, where all the patients were non-diabetic. Echocardiography did not show any wall motion abnormality and changes in the T wave and dysrhythmia were also not seen by ECG. 7 patients had high blood pressure, 11 had abnormal HDL, and 1 had abnormal LDL. In this study, serum LDL and HDL levels were not significantly correlated with age and BMI. Conversely, age and BMI were significantly associated with hypertension. Hypertension was observed in people over the age of 25 years and in overweighed individuals. 78.8% had normal BMI and 21.3% were overweighed. There was no significant correlation between serum LDL, serum HDL, and blood pressure and sex. The levels of abnormal LDL and HDL were higher in men than in women. ECG findings from our study did not report any significant cardiac abnormalities among hemophilic patients. Cardiovascular risk factors were not significantly correlated in these patients.


Analysis of sympathetic discharges during cardiogenic hypertensive chemoreflex.
Serotonin activates a cardiogenic hypertensive chemoreflex. During this reflex, sympathetic efferent neural discharges were examined in 10 anesthetized dogs. Thoracic nerves were identified, severed distally, and placed across electrodes. The efferent neurogram was recorded along with an electrocardiogram and blood pressure and stored on tape. Neurograms were later analyzed by a Lab 8/e computer with a Schmitt trigger. The resting tone and the elicited discharges of the multifiber preparations were plotted as poststimulus histograms. The integrals of the histograms were normalized. During the reflex, sympathetic efferent discharges averaged 4.9 (+/- 0.8) s in duration and the total discharge frequency was 199 (+/- 16)% of control tone. The evoked peak blood pressure response averaged 221/171 mmHg. Following autonomic blockade, the attenuated blood pressure response averaged 125/96 mmHg. The sympathetic efferent discharges then lasted 5.2 (+/- 0.7) s (NS). However, total discharge frequency (268 +/- 37% of control tone) was significantly greater (P less than 0.01) that before autonomic blockade. These data suggest that baroreceptors influence the rate of discharge and the total amount of neural traffic, but not the duration of the sympathetic response evoked by the cardiogenic hypertensive chemoreflex.


Native low-density lipoprotein uptake by macrophage colony-stimulating factor-differentiated human macrophages is mediated by macropinocytosis and micropinocytosis.
To examine the pinocytotic pathways mediating native low-density lipoprotein (LDL) uptake by human macrophage colony-stimulating factor-differentiated macrophages (the predominant macrophage phenotype in human atherosclerotic plaques). We identified the kinase inhibitor SU6656 and the Rho GTPase inhibitor toxin B as inhibitors of macrophage fluid-phase pinocytosis of LDL. Assessment of macropinocytosis by time-lapse microscopy revealed that both drugs almost completely inhibited macropinocytosis, although LDL uptake and cholesterol accumulation by macrophages were only partially inhibited (approximately 40%) by these agents. Therefore, we investigated the role of micropinocytosis in mediating LDL uptake in macrophages and identified bafilomycin A1 as an additional partial inhibitor (approximately 40%) of macrophage LDL uptake that targeted micropinocytosis. When macrophages were incubated with both bafilomycin A1 and SU6656, inhibition of LDL uptake was additive (reaching 80%), showing that these inhibitors target different pathways. Microscopic analysis of fluid-phase uptake pathways in these macrophages confirmed that LDL uptake occurs through both macropinocytosis and micropinocytosis. Our findings show that human macrophage colony-stimulating factor-differentiated macrophages take up native LDL by macropinocytosis and micropinocytosis, underscoring the importance of both pathways in mediating LDL uptake by these cells.


Vaccine preventable diseases in eastern Maharashtra--a hospital based analysis.
A retrospective study of hospital admissions of six vaccine preventable diseases from 1982-86 was carried out to save as a baseline data for future evaluation of vaccination programmes. Children between 1-4 years predominated the admissions in all diseases except whooping cough. The frequency of admissions of tuberculosis showed no change. Measles showed classical cyclical trend. Male female ratio was 1.8: 1.0. The mortality rate in meningeal tuberculosis was not influenced by the immunization status. Relatively small number of admissions of diphtheria and whooping cough indicates a decline due to natural causes or due to intervention programmes.


Differential expression of CuZn- and Fe-superoxide dismutase genes of tobacco during development, oxidative stress, and hormonal treatments.
Chloroplasts of Nicotiana tabacum have two superoxide dismutases: a Fe- and a CuZn-containing enzyme, encoded by the nuclear genes sodB and sodCp, respectively. As a first step in studying the physiological function of these two enzymes, we compared the expression of sodB and sodCp in different plant organs, in response to hormonal treatments, and upon treatment with paraquat and Norflurazon. The sodCp transcript and active enzyme were detected only in young leaves of mature plants. The sodB transcript was more abundant in young compared to old leaves, but the enzymatic activity was higher in mature and senescent leaves. sodCp and sodB exhibited a different expression pattern upon treatment with abscisic acid, indole-3-acetic acid, kinetin, gibberellin, and 1-aminocyclopropane-1-carboxylate. Paraquat treatment caused a decrease in abundance of both transcripts, although the dose dependency of this decrease differed. Norflurazon-induced photooxidation resulted in a 10-fold increase of sodCp mRNA whereas the sodB transcript level was 25% higher than the control. These differences in expression might explain why both plastid-located superoxide dismutase enzymes are needed, particularly under stress conditions.


Differences in Approach and Avoidance Motivation Sensitivities Predicting Participation and Performance in Strength Sport.
Cusimano, K, Freeman, P, Moran, J, and Yamaguchi, M. Differences in approach and avoidance motivation sensitivities predicting participation and performance in strength sport. J Strength Cond Res 38(1): 180-184, 2024-Gray's Reinforcement Sensitivity Theory proposes that individual differences in behavior are due to the sensitivity to 2 brain systems: the behavioral inhibition system (BIS), which regulates aversive emotions to threatening stimuli, and the behavioral approach system (BAS), which regulates positive emotions toward rewarding or nonpunishing stimuli. The current study investigated whether BIS and BAS sensitivity predicts participation and performance in strength sports. A sample of 177 competitive strength athletes (male = 148; female = 29; mean age = 28.68; SD = 6.24 years) and 178 control participants (male = 89; female = 89; mean age = 29.39; SD = 7.42) completed the BIS/BAS scale, with strength athletes also providing their Wilks scores as a measure of sporting performance. Independent t tests showed significantly higher BIS (MD = 2.37, p = 0.003, 95% CI [0.79, 3.94], d = 0.31) and total BAS (MD = 11.71, p < 0.001, 95% CI [9.26, 14.15], d = 1.00) sensitivity in strength athletes than individuals in the control group. A 3-step hierarchical regression analysis revealed that the number of training years (β = 0.506, p < 0 .001), BIS (β = -203, p = 0.005) and BAS drive (β = 0.188, p = 0.012) made significant unique contributions to predicting the Wilks score, with no significant contributions of age, sex, BAS fun seeking, and BAS reward responsiveness. The findings indicate that the overall reward sensitivity (total BAS score) and reward seeking (BAS drive) are associated positively with participation and performance in strength sports, respectively. Given the association of these brain systems to addiction and other psychiatric disorders, the findings could have implications in psychiatric treatment and sporting recruitment.


A prospective study of the accuracy of the surgeon's diagnosis and significance of positive margins in nonmelanoma skin cancers.
Even with a precise preoperative diagnosis, complete excision of nonmelanoma skin cancer is not always achieved. The conundrum remains the decision for appropriate secondary treatment. Many surgeons, regardless of the nature of the lesion, consider re-excision to be the only option. In a prior 4-year prospective study that ascertained the accuracy of our clinical diagnosis of skin lesions removed in an office setting, one-fifth were found to be malignant and 98 percent (n = 415) of the lesions were nonmelanoma skin cancer. Unfortunately, 65 (15.7 percent) of the malignant nonmelanoma skin cancer lesions had positive margins. The outcome of our management for these specific lesions was followed prospectively over the 7.5 years of this study to determine whether aggressive surgical intervention was justified in every case. Of 65 patients with lesions, early and complete re-excision of margin-positive nonmelanoma skin cancer was performed for 34 (52.3 percent), with residual tumor found in 11 (32.4 percent), followed by a later recurrence in one (2.9 percent). The remaining 31 patients agreed to semiannual office visits, with one (3.2 percent) recurrence in this group. Thus, the overall rate of recurrence for margin-positive nonmelanoma skin cancer was 3.1 percent, with a mean follow-up of 3.6 years (range, 0 to 7.5 years). There were no recurrences for basal cell carcinoma in either treatment group, suggesting that, at least for "simple" primary lesions without confounding risk factors, there is some validity to a "wait and see" attitude, in which treatment of a potential recurrence would be straightforward. Despite our observed infrequent local recurrences of squamous cell cancers (13.3 percent), the small risk of metastases still suggests the appropriateness of complete surgical eradication for these tumors whenever feasible.


Extracellular signals and receptor-like kinases regulating ROP GTPases in plants.
Rho-like GTPase from plants (ROPs) function as signaling switches that control a wide variety of cellular functions and behaviors including cell morphogenesis, cell division and cell differentiation. The Arabidopsis thaliana genome encodes 11 ROPs that form a distinct single subfamily contrarily to animal or fungal counterparts where multiple subfamilies of Rho GTPases exist. Since Rho proteins bind to their downstream effector proteins only in their GTP-bound "active" state, the activation of ROPs by upstream factor(s) is a critical step in the regulation of ROP signaling. Therefore, it is critical to examine the input signals that lead to the activation of ROPs. Recent findings showed that the plant hormone auxin is an important signal for the activation of ROPs during pavement cell morphogenesis as well as for other developmental processes. In contrast to auxin, another plant hormone, abscisic acid, negatively regulates ROP signaling. Calcium is another emerging signal in the regulation of ROP signaling. Several lines of evidence indicate that plasma membrane localized-receptor like kinases play a critical role in the transmission of the extracellular signals to intracellular ROP signaling pathways. This review focuses on how these signals impinge upon various direct regulators of ROPs to modulate various plant processes.


The bioflavonoid galangin suppresses the growth of ehrlich ascites carcinoma in Swiss Albino mice: a molecular insight.
Bioflavonoids are plant compounds touted for their potential to treat or prevent several diseases including cancer caused by various stress conditions. Galangin (4H-1-Benzopyran-4-one, 3, 5, 7-trihydroxy-2-phenyl-), a flavonoid, is a polyphenolic compound found primarily in medicinal herb, Alpinia galanga. This study aims to demonstrate the galangin as a pharmacological lead compound using in vitro, in vivo, and in silico model targeting specific cancer condition and proteins. The proliferation of MCF-7 and Ehrlich ascites carcinoma (EAC) cells was significantly inhibited with an IC₅₀ of 34.11 and 22.29 μg/ml, respectively. In an animal model system, galangin has inhibited the tumor growth by 73.51% ± 4.742 in EAC-induced Swiss Albino mice with no evidences of mortality as compared to standard drug, 5-fluorouracil. The effectiveness of galangin is proven in an animal system suggesting its pharmacokinetics behavior in an animal model which is also complemented by outcome of in silico analysis with more than 88 % of human intestinal absorption and significant Caco-2 cell, MDCK cell, and skin permeability as predicted by in silico methods. Galangin was docked against 19 different proteins involved in tumorogenesis and apoptosis; the energetic analysis indicates that it exhibits higher predicted binding free energy of -12.7 kcal/mol with Bcl-xL protein.


Patterns of endocrine reactivity in patients with recent myocardial infarction. Clinical and biochemical correlations: trial of endocrine therapy.
The following endocrine function parameters were studied serially in a group of 10 patients with recent myocardial infarction: blood and urinary levels of epinephrine and norepinephrine, urinary excretion of vanillyl-mandelic acid; protein-bound iodine, Hamolsky test (Hamolsky, Stein, and Freedberg, 1957); blood insulin; 24-hour urinary excretion of 17-hydroxycorticoids, sodium, and potassium. The acute phase of myocardial infarction, especially in those patients with a severe clinical course (rhythm disturbances, coronary insufficiency, circulatory failure), was associated with disturbed endocrine reactivity. The most frequent and the earliest feature was the increased level of the 24-hour urinary excretion of epinephrine, combined with a pronounced decrease in blood insulin level. Later in the course of the disease, as the adrenergic reactivity returned to normal, there was an increase in blood insulin to normal levels. In 3 patients with severe clinical symptoms of acute myocardial infarction, there were, in addition to the increased 24-hour urinary excretion of catecholamines, a decreased blood insulin, higher than normal levels of protein-bound iodine, and of the Hamolsky test. One of these patients developed hypoadrenia. It is possible that the abnormal endocrine reactions accelerate the catabolic processes within cardiac tissue (catecholamines, thyroid hormones), especially when there is a possible functional deficiency of hormones, occurring as a general adaptation reaction to stress (cortisol, insulin). The disturbances that follow may be dangerous for the patient.


High level expression and purification of antimicrobial human cathelicidin LL-37 in Escherichia coli.
The human antimicrobial peptide LL-37 is a cationic peptide with antimicrobial activity against both Gram-positive and Gram-negative microorganisms. This work describes the development of an expression system based on Escherichia coli capable of high production of the recombinant LL-37. The fusion protein Trx-LL-37 was expressed under control of T7 promoter. The expression of T7 polymerase in the E. coli strain constructed in this work was controlled by regulation mechanisms of the arabinose promoter. The expression plasmid was stabilized by the presence of parB locus which ensured higher homology of the culture during cultivation without antibiotic selection pressure. This system was capable of producing up to 1 g of fusion protein per 1 l of culture. The subsequent semipreparative HPLC allowed us to isolate 40 mg of pure LL-37. LL-37 showed high antimicrobial activity against both Gram-negative and Gram-positive microorganisms. Its activity against Candida albicans was practically nonexistent. Minimal Inhibition Concentration (MIC) determined for E. coli was 1.65 microM; for Staphylococcus aureus 2.31 microM, and for Enterococcus faecalis 5.54 microM. The effects of cathelicidin on E. coli included the ability to permeabilize both cell membranes, as could be observed by the increase of beta-galactosidase activity in extracellular space in time. Physiological changes were studied by scanning electron microscopy; Gram-positive microorganisms did not show any visible changes in cell shapes while the changes observed on E. coli cells were evident. The results of this work show that the herein designed expression system is capable of producing adequate quantities of active human antimicrobial peptide LL-37.


Immunological self, nonself discrimination.
The ability of immunodominant peptides derived from several antigen systems to compete with each other for T cell activation was studied. Only peptides restricted by a given transplantation antigen are mutually competitive. There is a correlation between haplotype restriction, ability to bind to the appropriate transplantation antigen, and ability to inhibit activation of other T cells restricted by the same transplantation antigen. An exception was noted in that a peptide derived from an antigen, bacteriophage lambda cI repressor, binds to the I-Ed molecule in a specific way, yet is not I-Ed-restricted. Comparison of the sequence of the repressor peptide with that of other peptides able to bind to (and be restricted by) I-Ed and a polymorphic region of the I-Ed molecule itself revealed a significant degree of homology. Thus, peptides restricted by a given class II molecule appear to be homologous to a portion of the class II molecule itself. The repressor-derived peptide is identical at several polymorphic residues at this site, and this may account for the failure of I-Ed to act as a restriction element. Comparison of antigenic peptide sequences with transplantation antigen sequences suggests a model that provides a basis for explaining self, nonself discrimination as well as alloreactivity.


UV mutagenesis for the overproduction of xylanase from Bacillus mojavensis PTCC 1723 and optimization of the production condition.
This study highlights xylanase overproduction from Bacillus mojavensis via UV mutagenesis and optimization of the production process. Bacillus mojavenis PTCC 1723 underwent UV radiation. Mutants' primary screening was based on the enhanced Hollow Zone Diameter/ Colony Diameter Ration (H/C ratios) of the colonies in comparison with the wild strain on Xylan agar medium. Secondly, enzyme production of mutants was compared with parental strain. Optimization process using lignocellulolytic wastes was designed with Minitab software for the best overproducer mutant. H/C ratio of 3.1 was measured in mutant number 17 in comparison with the H/C ratio of the parental strain equal to 1.6. Selected mutant produced 330.56 IU/ml xylanase. It was 3.45 times more enzyme than the wild strain with 95.73 IU/ml xylanase. Optimization resulted 575 IU/ml xylanase, with wheat bran as the best carbon source, corn steep liquor as the best nitrogen source accompanied with natural bakery yeast powder, in a medium with pH 7, after 48 hr incubation at 37°C, and the shaking rate of 230 rpm. Optimum xylanase activity was assayed at pH 7 and 40°C. Enzyme stability pattern shows it retains 62% of its initial activity at pH 9 after 3 hr. It also maintains up to 66% and 59% of its initial activity after 1 hr of pre-incubation at 70°C and 80°C. Mutation and optimization caused 5.9 times more enzyme yield by mutant strain. Also this enzyme can be categorized as an alkali-tolerant and thermo-stable xylanase.


The N-terminal TOG domain of Arabidopsis MOR1 modulates affinity for microtubule polymers.
Microtubule-associated proteins of the highly conserved XMAP215/Dis1 family promote both microtubule growth and shrinkage, and move with the dynamic microtubule ends. The plant homologue, MOR1, is predicted to form a long linear molecule with five N-terminal TOG domains. Within the first (TOG1) domain, the mor1-1 leucine to phenylalanine (L174F) substitution causes temperature-dependent disorganization of microtubule arrays and reduces microtubule growth and shrinkage rates. By expressing the two N-terminal TOG domains (TOG12) of MOR1, both in planta for analysis in living cells and in bacteria for in vitro microtubule-binding and polymerization assays, we determined that the N-terminal domain of MOR1 is crucial for microtubule polymer binding. Tagging TOG12 at the N-terminus interfered with its ability to bind microtubules when stably expressed in Arabidopsis or when transiently overexpressed in leek epidermal cells, and impeded polymerase activity in vitro. In contrast, TOG12 tagged at the C-terminus interacted with microtubules in vivo, rescued the temperature-sensitive mor1-1 phenotype, and promoted microtubule polymerization in vitro. TOG12 constructs containing the L174F mor1-1 point mutation caused microtubule disruption when transiently overexpressed in leek epidermis and increased the affinity of TOG12 for microtubules in vitro. This suggests that the mor1-1 mutant protein makes microtubules less dynamic by binding the microtubule lattice too strongly to support rapid plus-end tracking. We conclude from our results that a balanced microtubule affinity in the N-terminal TOG domain is crucial for the polymerase activity of MOR1.


Extracellular Matrix Molecular Remodeling in Human Liver Fibrosis Evolution.
Chronic liver damage leads to pathological accumulation of ECM proteins (liver fibrosis). Comprehensive characterization of the human ECM molecular composition is essential for gaining insights into the mechanisms of liver disease. To date, studies of ECM remodeling in human liver diseases have been hampered by the unavailability of purified ECM. Here, we developed a decellularization method to purify ECM scaffolds from human liver tissues. Histological and electron microscopy analyses demonstrated that the ECM scaffolds, devoid of plasma and cellular components, preserved the three-dimensional ECM structure and zonal distribution of ECM components. This method has been then applied on 57 liver biopsies of HCV-infected patients at different stages of liver fibrosis according to METAVIR classification. Label-free nLC-MS/MS proteomics and computation biology were performed to analyze the ECM molecular composition in liver fibrosis progression, thus unveiling protein expression signatures specific for the HCV-related liver fibrotic stages. In particular, the ECM molecular composition of liver fibrosis was found to involve dynamic changes in matrix stiffness, flexibility and density related to the dysregulation of predominant collagen, elastic fibers and minor components with both structural and signaling properties. This study contributes to the understanding of the molecular bases underlying ECM remodeling in liver fibrosis and suggests new molecular targets for fibrolytic strategies.


[Sympathetic ophthalmia versus Harada disease].
An algerian woman presents with a panuveitis of the right eye. Main features of this uveitis are exudative retinal detachments. That happened several months after a corneal ulcer of unknown aetiology on the left eye. Immunologic, neurologic and infections workup associated with previous ocular injury points to the diagnosis of sympathetic ophthalmia. However, the diagnosis of Harada disease can also be considered in this case because of similarities regarding the clinical and the histologic aspects which are exposed. If corticosteroids are the drug of choice, enucleation is still indicated for sympathetic ophthalmia.


Cyclin D1 is an early target in hepatocyte proliferation induced by thyroid hormone (T3).
The thyroid hormone (T3) affects cell growth, differentiation, and regulates metabolic functions via its interaction with the thyroid hormone nuclear receptors (TRs). The mechanism by which TRs mediate cell growth is unknown. To investigate the mechanisms responsible for the mitogenic effect of T3, we have determined changes in activation of transcription factors, mRNA levels of immediate early genes, and levels of proteins involved in the progression from G1 to S phase of the cell cycle. We show that hepatocyte proliferation induced by a single administration of T3 to Wistar rats occurred in the absence of activation of AP-1, NF-kappa B, and STAT3 or changes in the mRNA levels of the immediate early genes c-fos, c-jun, and c-myc. These genes are considered to be essential for liver regeneration after partial hepatectomy (PH). On the other hand, T3 treatment caused an increase in cyclin D1 mRNA and protein levels that occurred much more rapidly compared to liver regeneration after 2/3 PH. The early increase in cyclin D1 expression was associated with accelerated onset of DNA synthesis, as demonstrated by a 20-fold increase of bromodeoxyuridine-positive hepatocytes at 12 h after T3 treatment and by a 20-fold increase in mitotic activity at 18 h. An early increase of cyclin D1 expression was also observed after treatment with nafenopin, a ligand of a nuclear receptor (peroxisome proliferator-activated receptor alpha) of the same superfamily of steroid/thyroid receptors. T3 treatment also resulted in increased expression of cyclin E, E2F, and p107 and enhanced phosphorylation of pRb, the ultimate substrate in the pathway leading to transition from G1 to S phase. The results demonstrate that cyclin D1 induction is one of the earlier events in hepatocyte proliferation induced by T3 and suggest that this cyclin might be a common target responsible for the mitogenic activity of ligands of nuclear receptors.


WITHDRAWN: Home-based HIV voluntary counseling and testing in developing countries.
The low uptake of HIV voluntary counseling and testing (VCT), an effective HIV prevention intervention, has hindered global attempts to prevent new HIV infections, as well as limiting the scale-up of HIV care and treatment for the estimated 38 million infected persons. According to UNAIDS, only 10% of HIV-infected individuals worldwide are aware of their HIV status. At this point in the HIV epidemic, a renewed focus has shifted to prevention, and with it, a focus on methods to increase the uptake of HIV VCT. This review discusses home-based HIV VCT delivery models, which, given the low uptake of facility-based testing models, may be an effective avenue to get more patients on treatment and prevent new infections. (1) To identify and critically appraise studies addressing the implementation of home-based HIV voluntary counseling and testing in developing countries. (2) To determine whether home-based HIV voluntary counseling and testing (HBVCT) is associated with improvement in HIV testing outcomes compared to facility-based models. We searched online for published and unpublished studies in MEDLINE (February 2007), EMBASE (February 2007), CENTRAL (February 2007). We also searched databases listing conference proceedings and abstracts; AIDSearch (February 2007), The Cochrane Library (Issue 2, 2007), LILACS, CINAHL and Sociofile. We also contacted authors who have published on the subject of review. We searched for randomized controlled trials (RCTs) and non-randomized trials (e.g., cohort, pre/post-intervention and other observational studies) comparing home-based HIV VCT against other testing models. We independently selected studies, assessed study quality and extracted data. We expressed findings as odds ratios (OR), and relative Risk (RR) together with their 95% confidence intervals (CI). We identified one cluster-randomized trial and one pre/post-intervention (cohort) study, which were included in the review. An additional two ongoing RCTs were identified. All identified studies were conducted in developing countries. The two included studies comprised one cluster-randomized trial conducted in an urban area in Lusaka, Zambia and one pre/post-intervention (cohort) study, part of a rural community cohort in Southwestern Uganda. The two studies, while differing in methodology, found very high acceptability and uptake of VCT when testing and or results were offered at home, compared to the standard (facility-based testing and results). In the cluster-randomized trial (n=849), subjects randomized to an optional testing location (including home-based testing) were 4.6 times more likely to accept VCT than those in the facility arm (RR 4.6, 95% CI 3.6-6.2). Similarly, in the pre/post study (n=1868) offering participants the option of home delivery of results increased VCT uptake. In the intervention year (home delivery) participants were 5.23 times more likely to receive their results than during the year when results were available only at the facility. (OR 5.23 95% CI 4.02-6.8). Home-based testing and/or delivery of HIV test results at home, rather than in clinics, appears to lead to higher uptake in testing. However, given the limited extant literature and the limitations in the included existing studies, there is not sufficient evidence to recommend large-scale implementation of the home-based testing model.