Putting cells under pressure: a simple and efficient way to enhance the productivity of medium-chain-length polyhydroxyalkanoate in processes with Pseudomonas putida KT2440.
The success of bioprocess implementation relies on the ability to achieve high volumetric productivities and requires working with high-cell-density cultivations. Elevated atmospheric pressure might constitute a promising tool for enhancing the oxygen transfer rate (OTR), the major growth-limiting factor for such cultivations. However, elevated pressure and its effects on the cellular environment also represent a potential source of stress for bacteria and may have negative effects on product formation. In order to determine whether elevated pressure can be applied for enhancing productivity in the case of medium-chain-length polyhydroxyalkanoate (mcl-PHA) production by Pseudomonas putida KT2440, the impact of a pressure of 7 bar on the cell physiology was assessed. It was established that cell growth was not inhibited by this pressure if dissolved oxygen tension (DOT) and dissolved carbon dioxide tension (DCT) were kept below ∼30 and ∼90 mg L(-1), respectively. Remarkably, a little increase of mcl-PHA volumetric productivity was observed under elevated pressure. Furthermore, the effect of DCT, which can reach substantial levels during high-cell-density processes run under elevated pressure, was investigated on cell physiology. A negative effect on product formation could be dismissed since no significant reduction of mcl-PHA content occurred up to a DCT of ∼540 mg L(-1). However, specific growth rate exhibited a significant decrease, indicating that successful high-cell-density processes under elevated pressure would be restricted to chemostats with low dilution rates and fed-batches with a small growth rate imposed during the final part. This study revealed that elevated pressure is an adequate and efficient way to enhance OTR and mcl-PHA productivity. We estimate that the oxygen provided to the culture broth under elevated pressure would be sufficient to triple mcl-PHA productivity in our chemostat system from 3.4 (at 1 bar) to 11 g L(-1)h(-1) (at 3.2 bar).


Identifying Dysregulated Epigenetic Enzyme Activity in Castrate-Resistant Prostate Cancer Development.
There is a tremendous need for novel strategies aimed at directly assessing activities of histone modifiers to probe epigenetic determinants associated with disease progression. Here, we developed a high-throughput peptide microarray assay to identify altered histone lysine (de)acetylation activity in prostate cancer (PCa). This microarray-based activity assay revealed up-regulated histone acetyltransferase (HAT) activity against specific histone H3 sites in a castrate-resistant (CR) PCa cell line compared to its hormone-sensitive (HS) isogenic counterpart. NAD+-dependent deacetylation assays revealed down-regulated sirtuin activity in validated CR lines. Levels of acetyltransferases GCN5, PCAF, CBP, and p300 were unchanged between matched HS and CR cell lines. However, autoacetylation of p300 at K1499, a modification known to enhance HAT activity and a target of deacetylation by SIRT2, was highly elevated in CR cells, while SIRT2 protein level was reduced in CR cells. Interrogation of HS and matched CR xenograft lines reveals that H3K18 hyperacetylation, increased p300 activity, and decreased SIRT2 expression are associated with progression to CR in 8/12 (66%). Tissue microarray analysis revealed that hyperacetylation of H3K18 is a feature of CRPC. Inhibition of p300 results in lower H3K18ac levels and increased expression of androgen receptors. Thus, a novel histone array identifies altered enzyme activities during the progression to CRPC and may be utilized in a personalized medicine approach. Reduced SIRT2 expression and increased p300 activity lead to a concerted mechanism of hyperacetylation at specific histone lysine sites (H3K9, H3K14, and H3K18) in CRPC.


Your prodrug releases formaldehyde: should you be concerned? No!
The title of this commentary contains a frequently asked question whenever someone presents or proposes a prodrug strategy that releases formaldehyde as a result of bioconversion of a prodrug to parent drug. Formaldehyde, a highly water-soluble one-carbon molecule, is endogenous to cells, tissues, and body fluids. Although formaldehyde is generated and incorporated into essential metabolic processes by the human body, exposure to large amounts of formaldehyde vapor can irritate the nasal mucosa and may potentially be carcinogenic. It also gives a positive Ames test. Metabolism of both endogenous and exogenous formaldehyde involves rapid oxidation to formic acid catalyzed by glutathione dependent and independent dehydrogenases in the liver and erythrocytes. Balancing this rapid detoxification pathway is endogenous formation from normal metabolic processes and exogenous formaldehyde input, resulting in approximately 0.1 mM systemic levels. The possibility that formaldehyde released upon bioconversion of prodrugs might induce toxicity has been repeatedly stated, but no convincing evidence for this perceived toxicity has been documented in experimental studies. Therefore, as pharmaceutical chemists and not as toxicologists, we present our perspective on the apparent concern with release of formaldehyde as a by-product of in vivo bioconversion of selective prodrugs, and suggest that in comparison to the total amount of daily endogenous formaldehyde production from metabolism, and exogenous exposure from food and the environment, the amount generated by prodrugs is minute and is unlikely to cause any systemic toxicity in humans. Such an argument does not preclude formaldehyde-based toxicity assessment of a prodrug. Instead, it reduces the risk that in vivo liberation of formaldehyde will cause undue toxicity.


Full-length tobacco mosaic virus RNAs and defective RNAs have different 3' replication signals.
The viral replicase complex of positive-stranded RNA viruses interacts with cis-acting elements that are usually located at the termini of the viral RNAs. On comparison of the replication requirement of a tobacco mosaic virus (TMV)-based defective RNA (dRNA) and its helper virus, we found different requirements for replication of TMV RNAs in cis and in trans. The level of replication of full-length TMV RNA decreased substantially in the absence of pseudoknot (pk) 1 and/or 2, whereas identical deletions in dRNAs did not affect their replication. However, pk3 was required for replication of both full-length TMV RNAs and dRNAs. The requirements for homologous sequences were greater for dRNA replication than for replication of full-length TMV RNAs. Defective RNAs with heterologous 3' nontranslated regions (NTRs) failed to be replicated or replicated minimally, whereas replication of similarly mutated full-length RNAs was much less affected. Increasing amounts of contiguous heterologous sequences in the dRNAs compensated for the impaired interactions between the replicase and 3' NTR. The precision requirement appeared to involve the terminal 28 nucleotides, specifically the pseudoknot in the aminoacyl acceptor arm of the tRNA like structure, which was important in replication of both dRNAs and full-length TMV RNAs.


The American Pain Foundation TARGET Chronic Pain Initiative: better patient/clinician communication to improve pain management.
The American Pain Foundation has developed a program to improve clinician-chronic pain patient communication. The program includes a card describing a six part evaluation process for clinicians and patient notebook in which important daily information is recorded. The program is described including information on how to obtain the resources.


Lewis blood group determination in bloodstains by planimetric measurement of eluted monoclonal antibodies.
Planimetric measurements were employed for reading the results of an elution test to determine Lewis blood groups in dry human bloodstains. In the absorption-elution test, two varieties of indicators were used to detect eluted Lewis antibodies. First, 64 blood-stains aged between 2 to 8 months were tested with glutaraldehyde (GLA)-treated erythrocytes (planimetric hemagglutination assay, PMHA). This method demonstrated that dry stains weighing approximately 0.4 mg (equivalent to 3 microliters of whole blood) were sufficient for detection of Lea or Leb antigen. Results were obtained within 1 h. Then, 37 of these stains were tested with Lewis substance-coated latex particles (planimetric latex agglutination assay, PMLA). The presence of Lea and Leb antigen were detected from dry stains weighing 0.1 mg (equivalent to 1 microL of whole blood) within 3 h. Both these assays are faster and simpler with accuracy than the enzyme-linked immunosorbent assay (ELISA). Latex particles coated with Lewis substance are, in particular, strongly agglutinated and show agglutination patterns more clearly than erythrocytes. The blind tests using these two methods properly classified 7 Le(a + b-) and 23 Le(a-b + ) bloodstains; whereas, 5 Le(a-b-) stains were undetermined by the criteria for these tests. These results indicate the usefulness of the PMHA and PMLA for typing Lewis blood groups from small bloodstains.


Dysphagia following anterior cervical spinal surgery: a systematic review.
Dysphagia is a common complication of anterior surgery of the cervical spine. The incidence of post-operative dysphagia may be as high as 71% within the first two weeks after surgery, but gradually decreases during the following months. However, 12% to 14% of patients may have some persistent dysphagia one year after the procedure. It has been shown that female gender, advanced age, multilevel surgery, longer operating time and severe pre-operative neck pain may be risk factors. Although the aetiology remains unclear and is probably multifactorial, proposed causes include oesophageal retraction, prominence of the cervical plate and prevertebral swelling. Recently, pre-operative tracheal traction exercises and the use of retropharyngeal steroids have been proposed as methods of reducing post-operative dysphagia. We performed a systematic review to assess the incidence, aetiology, risk factors, methods of assessment and management of dysphagia following anterior cervical spinal surgery.


The Normal, the Radiosensitive, and the Ataxic in the Era of Precision Radiotherapy: A Narrative Review.
(1) Background: radiotherapy is a cornerstone of cancer treatment. When delivering a tumoricidal dose, the risk of severe late toxicities is usually kept below 5% using dose-volume constraints. However, individual radiation sensitivity (iRS) is responsible (with other technical factors) for unexpected toxicities after exposure to a dose that induces no toxicity in the general population. Diagnosing iRS before radiotherapy could avoid unnecessary toxicities in patients with a grossly normal phenotype. Thus, we reviewed iRS diagnostic data and their impact on decision-making processes and the RT workflow; (2) Methods: following a description of radiation toxicities, we conducted a critical review of the current state of the knowledge on individual determinants of cellular/tissue radiation; (3) Results: tremendous advances in technology now allow minimally-invasive genomic, epigenetic and functional testing and a better understanding of iRS. Ongoing large translational studies implement various tests and enriched NTCP models designed to improve the prediction of toxicities. iRS testing could better support informed radiotherapy decisions for individuals with a normal phenotype who experience unusual toxicities. Ethics of medical decisions with an accurate prediction of personalized radiotherapy's risk/benefits and its health economics impact are at stake; (4) Conclusions: iRS testing represents a critical unmet need to design personalized radiotherapy protocols relying on extended NTCP models integrating iRS.


The added value of ordinal analysis in clinical trials: an example in traumatic brain injury.
In clinical trials, ordinal outcome measures are often dichotomized into two categories. In traumatic brain injury (TBI) the 5-point Glasgow outcome scale (GOS) is collapsed into unfavourable versus favourable outcome. Simulation studies have shown that exploiting the ordinal nature of the GOS increases chances of detecting treatment effects. The objective of this study is to quantify the benefits of ordinal analysis in the real-life situation of a large TBI trial. We used data from the CRASH trial that investigated the efficacy of corticosteroids in TBI patients (n = 9,554). We applied two techniques for ordinal analysis: proportional odds analysis and the sliding dichotomy approach, where the GOS is dichotomized at different cut-offs according to baseline prognostic risk. These approaches were compared to dichotomous analysis. The information density in each analysis was indicated by a Wald statistic. All analyses were adjusted for baseline characteristics. Dichotomous analysis of the six-month GOS showed a non-significant treatment effect (OR = 1.09, 95% CI 0.98 to 1.21, P = 0.096). Ordinal analysis with proportional odds regression or sliding dichotomy showed highly statistically significant treatment effects (OR 1.15, 95% CI 1.06 to 1.25, P = 0.0007 and 1.19, 95% CI 1.08 to 1.30, P = 0.0002), with 2.05-fold and 2.56-fold higher information density compared to the dichotomous approach respectively. Analysis of the CRASH trial data confirmed that ordinal analysis of outcome substantially increases statistical power. We expect these results to hold for other fields of critical care medicine that use ordinal outcome measures and recommend that future trials adopt ordinal analyses. This will permit detection of smaller treatment effects.


Right ventricular apical contractility in acute pulmonary embolism: the McConnell sign revisited.
The McConnell sign has been regarded as a highly specific echo finding in acute pulmonary embolism (aPE). However, a completely satisfying explanation to account for this observation in aPE remains elusive. We used longitudinal velocity vector imaging (VVI) using a dedicated software program (Research Arena, Siemens, California) to assess regional right ventricular global and apical (RVa) mechanics between aPE and chronic pulmonary hypertension (cPH) patients. Standard echo parameters of RV performance as well as base to apex RV strain and dyssynchrony were quantified using VVI in a total of 30 patients. The population studied was divided into three groups: Group I included 10 healthy volunteers (50 +/- 16 years), Group II consisted of 10 patients (47 +/- 13 years) with known cPH, and Group III comprised 10 patients (58 +/- 18 years; P = 0.323) with documented aPE. Progressively lower indices of RV performance as well as RV basal and apical strain were recorded in Groups II and III, respectively. Most importantly, no difference in RVa segmental strain values was seen between Groups II and III. Based on this pilot data, aPE patients demonstrate a significant reduction in overall RV strain with a similar reduction in RV apical deformation. Therefore, regional RVa function is not truly spared in aPE and the probable visual illusion of preserved contractility might simply reflect tethering of the RVa to a hyperdynamic left ventricle in the presence of an acutely dilated RV.


Is Acupuncture Effective for Improving Insulin Resistance? A Systematic Review and Meta-analysis.
This study aimed to evaluate the clinical efficacy of acupuncture for intervening insulin resistance (IR) by meta-analysis of related randomized controlled trials (RCTs). Studies published prior to 31 January 2018 were searched on Pubmed, Medline, Cochrane Library, Embase databases and Chinese databases. Only RCTs, which examined acupuncture as the sole or adjunctive treatment for IR-related diseases, were included. The primary outcome was homeostasis model assessment for insulin resistance (HOMA-IR). The secondary outcomes consisted of fasting blood glucose (FBG), fasting insulin (FINS) and 2-h postprandial blood glucose (2h-PBG). The differences between groups were reported as mean differences (MD). All statistical analyses were performed using RevMan software 5.3. After carefully screening relevant studies, 9 RCTs involving 562 patients (279 in experimental group and 283 in control group) were enrolled in this study. The pooled results showed that acupuncture had significant effects on HOMA-IR (MD 0.70, 95% CI 0.04 to 1.35, P=0.04<0.05), FINS (MD 3.35 mU/L, 95% CI 1.99 to 4.7, P<0.001) and 2h-PBG (MD 1.03 mmol/L, 95% CI 0.25 to 1.82, P=0.01). However, the differences in FBG were not significant (MD 0.28 mmol/L, 95% CI-0.28 to 0.84, P=0.32>0.05). The present meta-analysis indicated that acupuncture can help to improve IR to a certain extent, which remains to be confirmed by further high-quality RCTs.


Atrial natriuretic factor in dogs with one-kidney, one-clip Goldblatt hypertension.
The temporal changes in the plasma concentration of immunoreactive atrial natriuretic factor (ANF) were studied in dogs (n = 5) before and after renal artery constriction to produce one-kidney, one-clip renovascular hypertension. Three days after renal artery constriction, circulating ANF increased from 34 +/- 7 to 344 +/- 129 pg/ml (P less than 0.05) in association with a 38-mmHg elevation of mean arterial pressure and increases in plasma renin and aldosterone levels. Plasma renin and aldosterone remained elevated for 9 days after renal artery constriction. Arterial pressure stabilized at this elevated level of 35-40 mmHg above base line for the duration of the 4 wk after renal artery constriction. Right atrial filling pressure did not change significantly (P greater than 0.05) after constriction of the renal artery. This observation suggests that volume expansion with right atrial distention is not the only stimulus for release of ANF in these hypertensive dogs. Plasma ANF remained significantly elevated above base-line levels for 7 days postconstriction even though a twofold decrease from the maximal response of 344 +/- 129 pg/ml on day 3 to 168 +/- 48 pg/ml on day 7 was observed during this same time interval. Plasma ANF continued to decrease toward prehypertensive levels throughout the remainder of the established phase of the hypertension lasting an additional 21 days of observation. The mechanisms for this observed pattern of plasma ANF are unclear but may involve changes in left atrial distention and stretch as a stimulus for hormone secretion with progression of the hypertension.


Gait alterations in the UAE population with and without diabetic complications using both traditional and entropy measures.
Diabetic foot, one of the most common and debilitating manifestations of type 2 diabetes mellitus (T2DM), is the leading cause of worldwide non-traumatic lower extremity amputations. Diabetics who are at risk of ulceration are currently mainly identified by a thorough clinical examination of the feet, which typically does not show clear symptoms during the early stages of disease progression. In this study, we used a non-linear dynamics tool, gait entropy (GaitEN), in addition to traditional linear gait analysis methods, to investigate gait alterations amongst diabetic patients with combinations of three types of T2DM related complications: retinopathy, diabetic peripheral neuropathy (DPN) and nephropathy. Peak plantar pressure (PPP) was not significantly different in the group with DPN as compared to the control group (diabetics with no complications, CONT) in the forefoot region (DPN: mean±SD: 396±69.4kPa, CONT: 409±68.9kPa), although it was significantly lower in the heel region (DPN: mean±SD: 285±43.1.4kPa, CONT: 295±61.8kPa). On the other hand, gait entropy was significantly lower for the DPN compared to CONT group (DPN: 0.95±0.34, CONT: 1.03±0.28, p<0.05). The significant low entropy was maintained when neuropathy was combined with either retinopathy or nephropathy. For the group with all three complications (ALL-C), the entropy was higher than CONT (ALL-C: 1.07±0.26). This may indicate an intrinsic sensorimotor feedback mechanism for the DPN patients to regulate their gait. However, this feedback gets weaker as patients develop multiple complications. Further analysis with longer walking time and different speeds is needed to verify the entropy results.


Cancer-specific loss of beta-defensin 1 in renal and prostatic carcinomas.
In a previous large-scale gene expression profiling study of renal epithelial neoplasms, human beta-defensin-1 (DEFB1) was found to be significantly down-regulated in conventional clear cell (renal) carcinoma. We have now completed an expanded expression analysis of this gene. We performed immunohistochemical analysis for the DEFB1 protein in clinical specimens of both renal cell carcinoma and prostate cancer. In a subset of prostate cancers, we performed laser capture microdissection and RT-PCR to correlate mRNA levels with protein levels. Overall, 82% of prostate cancers exhibit either complete loss of protein expression or only minimal expression, whereas the adjacent benign epithelium retained expression in all cases. Similarly, 90% of renal cell carcinomas show cancer-specific loss of DEFB1 protein. In the prostate cancer subset analysis, mRNA levels correlate with protein levels. We have thus demonstrated the cancer-specific down-regulation of DEFB1 in a large sample of prostatic and renal carcinomas and validated one of the key findings of previous cancer gene profiling studies of prostatic and renal neoplasia.


Go with the flow - bulk transport by molecular motors.
Cells are the smallest building blocks of all living eukaryotic organisms, usually ranging from a couple of micrometers (for example, platelets) to hundreds of micrometers (for example, neurons and oocytes) in size. In eukaryotic cells that are more than 100 µm in diameter, very often a self-organized large-scale movement of cytoplasmic contents, known as cytoplasmic streaming, occurs to compensate for the physical constraints of large cells. In this Review, we discuss cytoplasmic streaming in multiple cell types and the mechanisms driving this event. We particularly focus on the molecular motors responsible for cytoplasmic movements and the biological roles of cytoplasmic streaming in cells. Finally, we describe bulk intercellular flow that transports cytoplasmic materials to the oocyte from its sister germline cells to drive rapid oocyte growth.


Analysis of the microtubule-binding domain of MAP-2.
We examined the microtubule-binding domain of the microtubule-associated protein (MAP), MAP-2, using rabbit antibodies that specifically bind to the microtubule-binding region ("stub") and the projection portion ("arm") of MAP-2. We found that (a) microtubules decorated with arm antibody look similar to those labeled with whole unfractionated MAP antibody, though microtubules are not labeled with stub antibody; (b) incubation of depolymerized microtubule protein with stub antibody prior to assembly partially inhibits the rate of microtubule elongation, presumably because MAPs that are complexed with antibody cannot bind to microtubules and stabilize elongating polymers; (c) the rate of appearance and amounts of 36- and 40-kD microtubule-binding peptides produced by digestion with chymotrypsin are distinct for MAPs associated with microtubules vs. MAPs free in solution. The enhanced stability of the 40-kD peptide when associated with microtubules suggests that this domain of the protein is closely associated with, or partially buried in, the microtubule surface; (d) MAP-2 is a slender, elongate molecule as determined by unidirectional platinum shadowing (90 +/- 30 nm), which is in approximate agreement with previous observations. Stub antibody labels MAP-2 in the terminal one-quarter of the extended protein, indicating an intrinsic asymmetry in the molecule.


Inhibition of cyclin D1 expression and phosphorylation of retinoblastoma protein by phosmidosine, a nucleotide antibiotic.
In this report, we studied the effect of phosmidosine, a proline-containing nucleotide on the serum-induced cell cycle progression in human lung fibroblast WI-38 cells. Phosmidosine suppressed S-phase entry and arrested cell cycle progression at the G1 phase. In serum-stimulated cells, phosmidosine did not affect the activation of the mitogen-activated protein kinase cascade. However, phosmidosine inhibited hyperphosphorylation of retinoblastoma (RB) protein by RB-kinases such as cyclin-dependent kinase 4 and cyclin-dependent kinase 2, probably as a result of the inhibition of cyclin D1 expression. Furthermore, in tsFT210 cells, a temperature-sensitive cdc2 mutant isolated from the mouse mammary carcinoma cell line FM3A, phosmidosine, irreversibly inhibited the cell cycle progression at G1 without affecting the G2 to M transition. Phosmidosine acts at an earlier point in G1 compared with mimosine or aphidicolin, well-known cell cycle blockers at the G1-S boundary. Taken together, phosmidosine arrested cells at a specific point between the start point and restriction point in G1 and is a useful drug that may contribute to the understanding of the regulatory mechanisms of G1 progression.


A Dual-Function Hemicyanine Material with Highly Efficient Photothermal and Photodynamic Effect Used for Tumor Therapy.
Small molecular organic optical agents with synergistic effects of photothermal therapy (PTT) and photodynamic therapy (PDT), holds credible promise for anti-tumor therapy by overcoming individual drawbacks and enhanced photon utilization efficiency. However, developing effective dual-function PTT-PDT photosensitizers (PSs) for efficient synergistic phototherapy remains challenging. Here we present a benz[c,d]indolium-substituted hemicyanine named Rh-BI, which possesses a high photothermal conversion efficiency of 41.67% by exhaustively suppressing fluorescence emission. Meanwhile, the rotating phenyl group at meso-site induced charge recombination to enhance the molar extinction coefficient up to 13.58×104 M-1 cm-1 , thereby potentiating the photodynamic effect. Under 808 nm irradiation, Rh-BI exhibited significant phototoxicity in several cancer cell types in vitro with IC50 values as low as about 0.5 μM. Moreover, treatment of 4T1 tumor-bearing mice with Rh-BI under laser irradiation successfully inhibits tumor growth. In a word, we developed an effective strategy to build PTT-PDT dual-functional optical materials based on hemicyanine backbone for tumor therapy by modulating conjugation system interaction to adjust the energy consumption pathway. This article is protected by copyright. All rights reserved.


Macrophages in the cochlea; an immunological link between risk factors and progressive hearing loss.
Macrophages are abundant in the cochlea; however, their role in hearing loss is not well understood. Insults to the cochlea, such as noise or insertion of a cochlear implant, cause an inflammatory response, which includes activation of tissue-resident macrophages. Activation is characterized by changes in macrophage morphology, mediator expression, and distribution. Evidence from other organs shows activated macrophages can become primed, whereby subsequent insults cause an elevated inflammatory response. Primed macrophages in brain pathologies respond to circulating inflammatory mediators by disproportionate synthesis of inflammatory mediators. This signaling occurs behind an intact blood-brain barrier, similar to the blood-labyrinth barrier in the cochlea. Local tissue damage can occur as the result of mediator release by activated macrophages. Damage is typically localized; however, if it is to structures with limited ability to repair, such as neurons or hair cells within the cochlea, it is feasible that this contributes to the progressive loss of function seen in hearing loss. We propose that macrophages in the cochlea link risk factors and hearing loss. Injury to the cochlea causes local macrophage activation that typically resolves. However, in susceptible individuals, some macrophages enter a primed state. Once primed, these macrophages can be further activated, as a consequence of circulating inflammatory molecules associated with common co-morbidities. Hypothetically, this would lead to further cochlear damage and loss of hearing. We review the evidence for the role of tissue-resident macrophages in the cochlea and propose that cochlear macrophages contribute to the trajectory of hearing loss and warrant further study.


Global and local environmental changes as drivers of Buruli ulcer emergence.
Many emerging infectious diseases are caused by generalist pathogens that infect and transmit via multiple host species with multiple dissemination routes, thus confounding the understanding of pathogen transmission pathways from wildlife reservoirs to humans. The emergence of these pathogens in human populations has frequently been associated with global changes, such as socio-economic, climate or biodiversity modifications, by allowing generalist pathogens to invade and persist in new ecological niches, infect new host species, and thus change the nature of transmission pathways. Using the case of Buruli ulcer disease, we review how land-use changes, climatic patterns and biodiversity alterations contribute to disease emergence in many parts of the world. Here we clearly show that Mycobacterium ulcerans is an environmental pathogen characterized by multi-host transmission dynamics and that its infectious pathways to humans rely on the local effects of global environmental changes. We show that the interplay between habitat changes (for example, deforestation and agricultural land-use changes) and climatic patterns (for example, rainfall events), applied in a local context, can lead to abiotic environmental changes and functional changes in local biodiversity that favor the pathogen's prevalence in the environment and may explain disease emergence.


Etv5a regulates the proliferation of ventral mesoderm cells and the formation of hemato-vascular derivatives.
Hematopoietic and vascular endothelial cells constitute the circulatory system and are both generated from the ventral mesoderm. However, the molecules and signaling pathways involved in ventral mesoderm formation and specification remain unclear. We found that zebrafish etv5a was expressed in the ventral mesoderm during gastrulation. Knockdown of Etv5a using morpholinos increased the proliferation of ventral mesoderm cells and caused defects in hematopoietic derivatives and in vascular formation. By contrast, the formation of other mesodermal derivatives, such as pronephros, somites and the gut wall, was not affected. Knockdown specificity was further confirmed by overexpression of an etv5a construct lacking its acidic domain. In conclusion, our data reveal that etv5a is essential for the inhibition of ventral mesoderm cell proliferation and for the formation of the hemato-vascular lineage.


Diversification of the ADH1B gene during expansion of modern humans.
A variant allele, ADH1B*48His, also known as ADH1B*2, at the human Alcohol Dehydrogenase 1B gene (ADH1B) is strongly associated with alcoholism in some populations and has an unusual geographic distribution. Strong evidence implies selection has increased the frequency of this allele in some East Asian populations but does not fully explain its geographic pattern. We have studied haplotypes of 10 single nucleotide polymorphisms (SNPs) and two short tandem repeat polymorphisms (STRPs) in the ADH1B region in 2,206 individuals from a worldwide set of populations. These SNPs and STRPs define nine common haplogroups most of which have distinct geographic patterns. The haplogroups H5 and H6, both with the derived ADH1B*48His allele, appear restricted to the Middle East and East Asia, respectively. The positively selected H7 is derived from H6 by a new regulatory region variant defining SNP rs3811801 restricted to East Asia. Age estimates of the haplogroups based on the STRPs also agree with the time of the migration events estimated by other studies. H7 is estimated to have expanded recently, around 2,800 years ago, and ancient DNA samples from North China confirm its presence about that time. The dating of the H7 expansion may help understand the selective force on the ADH1B gene.


Angular Displacement and Velocity Sensors Based on Coplanar Waveguides (CPWs) Loaded with S-Shaped Split Ring Resonators (S-SRR).
In this paper, angular displacement and angular velocity sensors based on coplanar waveguide (CPW) transmission lines and S-shaped split ring resonators (S-SRRs) are presented. The sensor consists of two parts, namely a CPW and an S-SRR, both lying on parallel planes. By this means, line-to-resonator magnetic coupling arises, the coupling level being dependent on the line-to-resonator relative angular orientation. The line-to-resonator coupling level is the key parameter responsible for modulating the amplitude of the frequency response seen between the CPW ports in the vicinity of the S-SRR fundamental resonance frequency. Specifically, an amplitude notch that can be visualized in the transmission coefficient is changed by the coupling strength, and it is characterized as the sensing variable. Thus, the relative angular orientation between the two parts is measured, when the S-SRR is attached to a rotating object. It follows that the rotation angle and speed can be inferred either by measuring the frequency response of the S-SRR-loaded line, or the response amplitude at a fixed frequency in the vicinity of resonance. It is in addition shown that the angular velocity can be accurately determined from the time-domain response of a carrier time-harmonic signal tuned at the S-SRR resonance frequency. The main advantage of the proposed device is its small size directly related to the small electrical size of the S-SRR, which allows for the design of compact angular displacement and velocity sensors at low frequencies. Despite the small size of the fabricated proof-of-concept prototype (electrically small structures do not usually reject signals efficiently), it exhibits good linearity (on a logarithmic scale), sensitivity and dynamic range.


Image-guided positioning in intracranial non-invasive stereotactic radiosurgery for the treatment of brain metastasis.
The aim of the study was to examine the feasibility of non-invasive image-guided radiosurgery to improve patient comfort and quality of life in stereotactic radiosurgery planning and treatment of patients with brain metastasis. Precise immobilization is a rule of thumb for stereotactic radiosurgery. Non-invasive immobilization techniques have the potential of improved quality of life compared with invasive procedures. A total of 92 lesions from 42 patients with brain metastasis were included in the study. After immobilization with a thermoplastic mask and a bite-block unlike the invasive frame-based procedure, planning computed tomography images were acquired and fused with magnetic resonance images. After contouring, intensity-modulated stereotactic radiosurgery (IM-SRS) planning was done, and the patients were re-immobilized on the treatment couch for the therapy procedures. While patients were on the treatment couch, kilovoltage-cone beam computed tomography images were acquired to determine setup errors and achieve on-line correction and then repeated after on-line correction to confirm precise tumor localization. The patients then underwent single-fraction definitive treatment. For the 92 lesions treated, mean ± SD values of translational setup corrections in X (lateral), Y (longitudinal), and Z (vertical) dimensions were 0.7 ± 0.7 mm, 0.8 ± 0.7 mm, and 0.6 ± 0.5 mm, and rotational set-up corrections were 0.5 ± 1.1°, 0.06 ± 1.1°, and -0.1 ± 1.1° in X (pitch), Y (roll), and Z (yaw), respectively. The mean three-dimensional correction vector was 1.2 ± 1.1 mm. Non-invasive image-guided radiosurgery for brain metastasis is feasible, and the non-invasive treatment approach can be routinely used in clinical practice to improve patients' quality of life.


Patient-centred orientation of students from different healthcare disciplines, their understanding of the concept and factors influencing their development as patient-centred professionals: a mixed methods study.
A patient-centred approach to care is increasingly the mandate for healthcare delivery. There is a need to explore how health professional students develop patient-centred attributes. This study aims to understand the extent of patient-centred orientations of health professional students, their perceptions and factors influencing their adoption of the approach. The study used a cross-sectional, parallel mixed methods design combining a survey using the Patient-Practitioner Orientation Scale (PPOS) followed by focus groups with medical, nursing, physiotherapy and speech and language therapy students. Data included students' age, gender, programme, and placements experienced. Pearson's chi squared and the non-parametric equivalent Kruskal-Wallis H test were done to test for differences in demographics for appropriate variables. One-way ANOVA or Welch test was done to explore differences in PPOS scores. Regression analysis was done to test the influence of the demographic variables on PPOS scores. Data from focus groups were coded, categorised and organised under themes appropriate to the research aims. Of the 211 complete responses, significant differences were observed between medical and physiotherapy students in total PPOS scores, (MD -8.11 [95% CI -12.02 - 4.20] p = 0.000), Caring component (MD -4.44 [95% CI - 6.69, - 2.19] p = 0.000) and Sharing component (MD -3.67 [95% CI -6.12 -1.22] p = 0.001). The programme in which students were enrolled i.e. Medicine and SALT were the only indicators of higher PPOS total scores (F = 4.6 Df 10,69; p = 7.396e-06) and caring scores (F = 2.164 Df 10, 69 p = 0.022). Focus groups revealed that students perceived patient-centredness as holistic yet individualised care through establishing a partnership with patient. They identified that their student status, placement pressures, placement characteristics especially mentoring influenced their development of patient-centred attributes. This study highlights the fact that the pressures of training in the National Health Service affects the development of students' patient-centred orientation. There is a need for further work to explore aspects related to mentor training, for the development of patient-centred attributes, in a curricular framework structured on students' needs from this study.


Molecular profiling in lung cancer associated with cystic airspaces.
'Lung cancer associated with cystic airspaces' is a rare radiological entity that is more frequently encountered on imaging studies and is gaining more attention since the widespread use of CT for lung cancer screening. Numerous aspects of this entity remain unraveled, including molecular profiling. The goal of this observational retrospective single-center case series is to investigate the molecular profile of lung cancers presenting with this specific morphology in a Caucasian population. Between January 2014 and May 2017, 13 patients were presented at the Multidisciplinary Thoracic Oncology Tumor Board with imaging findings consistent with 'lung cancer associated with cystic airspaces'. Electronic medical files were reviewed for patient characteristics, stage, histopathological findings and - in particular - molecular profiling. Histopathological diagnosis showed adenocarcinoma in 11 patients in our series. Mutational analysis in 10 showed different molecular alterations: an EGFR exon 18 mutation, ROS1 rearrangement and BRAF mutation in one patient each. Two patients showed KRAS mutations. With 5 out of 10 patients with an adenocarcinoma presenting with cystic airspace morphology showing a molecular alteration, this may indicate that in this subgroup, molecular profiling is mandatory, regardless of smoking history.


Occurrence and Type of Adverse Events During the Use of Stationary Gait Robots-A Systematic Literature Review.
Robot-assisted gait training (RAGT) devices are used in rehabilitation to improve patients' walking function. While there are some reports on the adverse events (AEs) and associated risks in overground exoskeletons, the risks of stationary gait trainers cannot be accurately assessed. We therefore aimed to collect information on AEs occurring during the use of stationary gait robots and identify associated risks, as well as gaps and needs, for safe use of these devices. We searched both bibliographic and full-text literature databases for peer-reviewed articles describing the outcomes of stationary RAGT and specifically mentioning AEs. We then compiled information on the occurrence and types of AEs and on the quality of AE reporting. Based on this, we analyzed the risks of RAGT in stationary gait robots. We included 50 studies involving 985 subjects and found reports of AEs in 18 of those studies. Many of the AE reports were incomplete or did not include sufficient detail on different aspects, such as severity or patient characteristics, which hinders the precise counts of AE-related information. Over 169 device-related AEs experienced by between 79 and 124 patients were reported. Soft tissue-related AEs occurred most frequently and were mostly reported in end-effector-type devices. Musculoskeletal AEs had the second highest prevalence and occurred mainly in exoskeleton-type devices. We further identified physiological AEs including blood pressure changes that occurred in both exoskeleton-type and end-effector-type devices. Training in stationary gait robots can cause injuries or discomfort to the skin, underlying tissue, and musculoskeletal system, as well as unwanted blood pressure changes. The underlying risks for the most prevalent injury types include excessive pressure and shear at the interface between robot and human (cuffs/harness), as well as increased moments and forces applied to the musculoskeletal system likely caused by misalignments (between joint axes of robot and human). There is a need for more structured and complete recording and dissemination of AEs related to robotic gait training to increase knowledge on risks. With this information, appropriate mitigation strategies can and should be developed and implemented in RAGT devices to increase their safety.


A minimal kinetic model for the interpretation of complex catalysis in single enzyme molecules.
Multi-exponential waiting-time distribution and randomness parameter greater than unity ascribe dynamic disorder in single-enzyme catalysis corroborated to the interplay of transforming conformers [English et al., Nat. Chem. Biol., 2006, 2, 87]. The associated multi-state model of enzymatic turnovers with statically heterogeneous catalytic rates misdescribes the non-linear uprising of the randomness parameter from unity in relation to the attributes of the fall-offs of the waiting-time distribution at different substrate concentrations. To resolve this crucial issue, we first employ a comprehensive stochastic reaction scenario and further rationalize and work out the minimal indispensable dynamic-disorder model that ensures the foregoing relationship upon comparison with the data. We elucidate that specific disregard for the transition rate coefficients in the multi-state model on account of the especially slow conformational transitions is the underlying reason for not achieving interrelation between the observables.


Enhanced bioavailability of danazol nanosuspensions by wet milling and high-pressure homogenization.
The majority of drugs obtained through synthesis and development show poor aqueous solubility and dissolution velocity, resulting in reduced bioavailability of drugs. Most of these problems arise from formulation-related performance issues, and an efficient way to overcome these obstacles and to increase dissolution velocity is to reduce the particle size of drug substances to form drug nanosuspensions. Danazol nanosuspensions were prepared by wet milling (WM) and high-pressure homogenization (HPH) methods. The nanosuspensions obtained using these fabrication methods were analyzed for their particle size, surface charge, and the crystallinity of the product was assessed by X-ray diffraction (XRD) and differential scanning calorimetry techniques. To determine in vitro and in vivo performances of the prepared nanosuspensions, dissolution velocity, and bioavailability studies were performed. Particle size and zeta potential analysis showed the formation of nanosized particles with a negative charge on the surface. XRD depicted the nanocrystalline nature of danazol with low diffraction intensities. With increased surface area and saturation solubility, the nanosuspensions showed enhanced dissolution velocity and oral bioavailability in rats when compared to the bulk danazol suspension. The results suggest that the preparation of nanosuspensions by WM or HPH is a promising approach to formulate new drugs or to reformulate existing drugs with poorly water-soluble properties.


Extrusion and death of DPP/BMP-compromised epithelial cells in the developing Drosophila wing.
During animal development, epithelial cell fates are specified according to spatial position by extracellular signaling pathways. Among these, the transforming growth factor beta/bone morphogenetic protein (TGF-beta/BMP) pathways are evolutionarily conserved and play crucial roles in the development and homeostasis of a wide range of multicellular tissues. Here we show that in the developing Drosophila wing imaginal epithelium, cell clones deprived of the BMP-like ligand Decapentaplegic (DPP) do not die as previously thought but rather extrude from the cell layer as viable cysts exhibiting marked abnormalities in cell shape and cytoskeletal organization. We propose that in addition to assigning cell fates, a crucial developmental function of DPP/BMP signaling is the position-specific control of epithelial architecture.


Surgery for fibrosing mediastinitis with severe pulmonary hypertension due to pulmonary venous stenosis.
A 21-year-old woman with severe pulmonary hypertension and circulatory collapse was referred to our hospital for possible lung transplantation with extracorporeal membrane oxygenation support. Computed tomography revealed severe stenosis of all 4 pulmonary veins, and fibrosing mediastinitis was suspected. Surgical reconstruction of the pulmonary veins was performed, and extracorporeal membrane oxygenation support was weaned off. After surgery, pulmonary vascular resistance normalized. This successful case demonstrates that surgical pulmonary venous reconstruction is an important treatment for fibrosing mediastinitis induced by pulmonary venous stenosis and pulmonary hypertension.


NOMePlot: analysis of DNA methylation and nucleosome occupancy at the single molecule.
Recent technical advances highlight that to understand mammalian development and human disease we need to consider transcriptional and epigenetic cell-to-cell differences within cell populations. This is particularly important in key areas of biomedicine like stem cell differentiation and intratumor heterogeneity. The recently developed nucleosome occupancy and methylome (NOMe) assay facilitates the simultaneous study of DNA methylation and nucleosome positioning on the same DNA strand. NOMe-treated DNA can be sequenced by sanger (NOMe-PCR) or high throughput approaches (NOMe-seq). NOMe-PCR provides information for a single locus at the single molecule while NOMe-seq delivers genome-wide data that is usually interrogated to obtain population-averaged measures. Here, we have developed a bioinformatic tool that allow us to easily obtain locus-specific information at the single molecule using genome-wide NOMe-seq datasets obtained from bulk populations. We have used NOMePlot to study mouse embryonic stem cells and found that polycomb-repressed bivalent gene promoters coexist in two different epigenetic states, as defined by the nucleosome binding pattern detected around their transcriptional start site.


Automated and robust PERCIST-based thresholding framework for whole body PET-CT studies.
Positron emission tomography (PET) is unique for quantitatively assessing treatment response before marked morphological changes are detectable by Computed Tomography (CT). PET response criterion (PERCIST) is a widely accepted approach of assessing metabolic response of malignant lesions by using Standardized uptake value (SUV) normalized by lean body mass (LBM) with a volume of interest (VOI) reference defined in the right lobe of liver. However, the operator-dependent delineation of VOI reference is a time consuming and subjective task. Although the VOI reference can be estimated from the co-aligned CT, the low-dose CT data in PET-CT poses challenge in liver segmentation. In this study, we propose a fully automatic framework to calculate the PERCIST-based thresholding for whole-body PET-CT studies. The framework consists of multi-atlas registration and voxel classification for CT data to segment liver structure and delineate the VOI reference, which is then mapped to the PET data to derive the value of SUVLBM thresholding for PET to select regions of high metabolism. We evaluated our framework with 28 clinical studies diagnosed with lung cancer or lymphoma, and demonstrated both reliability and efficiency in depicting lesions using PERCIST thresholding.


Prevention and care of paediatric HIV infection in Ouagadougou, Burkina Faso: knowledge, attitudes and practices of the caregivers.
The paediatric Human Immunodeficiency Virus (HIV) epidemic still progresses because of operational challenges in implementing prevention of mother-to-child HIV transmission (PMCT) programs. We assessed the knowledge, attitudes and practices (KAP) of children's caregivers regarding mother-to-child transmission (MTCT) of HIV, paediatric HIV infection, early infant diagnosis (EID), and paediatric antiretroviral treatment in Ouagadougou, Burkina Faso. We undertook a qualitative survey in the four public hospitals managing HIV exposed or infected children, in Ouagadougou in 2011. A sociologist used a semi-structured questionnaire to interview caregivers of children less than 5 years old attending the paediatrics wards on their KAP. Study participants were divided into four groups as follows: those who did not yet know their children's HIV infection status, those who were waiting for their children's HIV test results, those who were waiting for antiretroviral treatment, and those who were already on antiretroviral treatment. A total of 37 caregivers were interviewed. The mean age was 32.5 years, and 29 (78 %) were mothers. Twenty seven (73 %) caregivers had primary or higher level of education, and 15 (40 %) described their occupation as "housewife". Overall, 36 (97 %) of caregivers knew that the main route of HIV transmission for infants was through MTCT and 14 (38 %) specified that it occurred during pregnancy or delivery. Five percent thought that MTCT of HIV occurred during conception. PMTCT interventions could help prevent infant HIV infection according to 32 (87 %) caregivers. Thirty five percent of caregivers stated EID as a prevention strategy. Fifty-four percent of the participants believed that replacement feeding option would prevent MTCT of HIV; 24 (65 %) stated that they would prefer medical practitioners seek caregivers' consent before carrying out any HIV-test for their child, and that caregivers' consent was not compulsory before antiretroviral treatment. All caregivers thought that it was necessary to treat HIV-infected children, although they did not know what interventions could be done. This study highlighted the low level of caregivers' knowledge on paediatric HIV prevention and care in Ouagadougou. Awareness programs targeting caregivers need to be strengthened in order to improve the uptake of HIV early infant diagnosis and care.


[Focus on GIST management].
Since the discovery of the remarkable efficacy of imatinib in the metastatic GIST, several studies advanced our knowledge on the care of this pathology. In the localized GIST, the efficacy of the adjuvant treatment by imatinib was proved, but the duration, the indication and the management in case of relapse after imatinib are not still consensual. The imatinib is also used in neoadjuvant setting to optimize the quality of resection, the main treatment remaining the maximal tumor resection. In metastatic setting, imatinib remains the standard of care first-line treatment. It must be administered until progress or intolerance. Nevertheless, secondary resistance to imatinib is a substantial problem in routine clinical practice; in second line, sunitinib demonstrated its efficacy. Several inhibitors of tyrosine-kinases are ongoing evaluated in all the therapeutic lines. Clearly, a better knowledge of the molecular profile and the pharmacokinetics underlying the resistance to imatinib as well as the development of a new class of broad-spectrum tyrosine-kinase inhibitors may allow in the near future new individualized therapeutic strategies for GIST patients.


aguA, the gene encoding an extracellular alpha-glucuronidase from Aspergillus tubingensis, is specifically induced on xylose and not on glucuronic acid.
An extracellular alpha-glucuronidase was purified and characterized from a commercial Aspergillus preparation and from culture filtrate of Aspergillus tubingensis. The enzyme has a molecular mass of 107 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 112 kDa as determined by mass spectrometry, has a determined pI just below 5.2, and is stable at pH 6.0 for prolonged times. The pH optimum for the enzyme is between 4.5 and 6.0, and the temperature optimum is 70 degrees C. The alpha-glucuronidase is active mainly on small substituted xylo-oligomers but is also able to release a small amount of 4-O-methylglucuronic acid from birchwood xylan. The enzyme acts synergistically with endoxylanases and beta-xylosidase in the hydrolysis of xylan. The enzyme is N glycosylated and contains 14 putative N-glycosylation sites. The gene encoding this alpha-glucuronidase (aguA) was cloned from A. tubingensis. It consists of an open reading frame of 2,523 bp and contains no introns. The gene codes for a protein of 841 amino acids, containing a eukaryotic signal sequence of 20 amino acids. The mature protein has a predicted molecular mass of 91,790 Da and a calculated pI of 5.13. Multiple copies of the gene were introduced in A. tubingensis, and expression was studied in a highly overproducing transformant. The aguA gene was expressed on xylose, xylobiose, and xylan, similarly to genes encoding endoxylanases, suggesting a coordinate regulation of expression of xylanases and alpha-glucuronidase. Glucuronic acid did not induce the expression of aguA and also did not modulate the expression on xylose. Addition of glucose prevented expression of aguA on xylan but only reduced the expression on xylose.


Properties of the FhuA channel in the Escherichia coli outer membrane after deletion of FhuA portions within and outside the predicted gating loop.
Escherichia coli transports Fe3+ as a ferrichrome complex through the outer membrane in an energy-dependent process mediated by the FhuA protein. A FhuA deletion derivative lacking residues 322 to 355 (FhuA delta322-355) forms a permanently open channel through which ferrichrome diffused. This finding led to the concept that the FhuA protein forms a closed channel that is opened by input of energy derived from the electrochemical potential across the cytoplasmic membrane, mediated by the Ton system. In this study, we constructed various FhuA derivatives containing deletions inside and outside the gating loop. FhuA delta322-336 bound ferrichrome and displayed a residual Ton-dependent ferrichrome transport activity. FhuA delta335-355 no longer bound ferrichrome but supported ferrichrome diffusion through the outer membrane in the absence of the Ton system. FhuA delta335-355 rendered cells sensitive to sodium dodecyl sulfate and supported diffusion of maltotetraose and maltopentaose in a lamB mutant lacking the maltodextrin-specific channel in the outer membrane. Cells expressing FhuA delta70-223, which has a large deletion outside the gating loop, were highly sensitive to sodium dodecyl sulfate and grew on maltodextrins but showed only weak ferrichrome uptake, suggesting formation of a nonspecific pore through the outer membrane. FhuA delta457-479 supported Ton-dependent uptake of ferrichrome. None of these FhuA deletion derivatives formed pores in black lipid membranes with a stable single-channel conductance. Rather, the conductance displayed a high degree of current noise, indicating a substantial influence of the deletions on the conformation of the FhuA protein. FhuA also supports infection by the phages T1, T5, and phi80 and renders cells sensitive to albomycin and colicin M. Cells expressing FhuA delta322-336 were sensitive to albomycin and colicin M but were only weakly sensitive to T5 and phi480 and insensitive to T1. Cells expressing FhuA delta335-355 were resistant to all FhuA ligands. These results indicate different structural requirements within the gating loop for the various FhuA ligands. Cells expressing FhuA delta457-479 displayed a strongly reduced sensitivity to all FhuA ligands, while cells expressing FhuA delta70-223 were rather sensitive to all FhuA ligands except albomycin, to which they were nearly resistant. It is concluded that residues 335 to 355 mainly determine the properties of the gate with regard to FhuA permeability and ligand binding.


[Intervening factors in attention flow of professionals injured by biological material].
To describe the barriers and facilitator factors to follow the attention flow of professionals injured by biological material in the worker perspective. Qualitative descriptive study with data collected through individual interviews with 18 injured workers, assisted in reference public units in the city of Goiânia. The content analysis was carried out with assistance of the ATLAS.ti 6.2 software, under the work organization and subjective perspectives. From the interviews regarding the barriers and facilitator factors emerged the categories: organizational structure, Support from close people, and Knowledge influence. The organized services have enabled more qualified consultations and the workers follow-up, which caused a satisfaction feeling in relation to the working environment.


A Case of Large Intraoral Teratoma with Intracranial Extension: Peri‑Operative Challenges.
A 40-day-old female child presented to us with an oral cavity mass, noticed at birth and progressively increasing in size. Preoperative computed tomography revealed intracranial extension of the lesion. Airway management, securing and maintaining invasive vascular access peri-operatively and transport of the patient to MRI suite were the major challenges. Mask ventilation was difficult and successful intubation required three attempts. Otorhinolaryngologists were present in the theatre during induction to perform a tracheostomy if the situation so demanded.


High temperature limits on developmental canalization in the ascidian Ciona intestinalis.
The normal embryogenesis of marine animals is typically confined to a species-specific range of temperatures. Within that temperature range development results in a consistent, or canalized, phenotype, whereas above and below the range abnormal phenotypes are produced. This study reveals a high temperature threshold, occurring over a 1-2 °C range, for normal embryonic development in C. intestinalis. Above that threshold the prevalence of morphological abnormalities increases significantly, beginning with cleavage and gastrula stages, and becoming more pronounced as embryogenesis proceeds. However, even in highly morphologically abnormal temperature disrupted (TD) embryos, muscle, endoderm, notochord, epidermis, and sensory pigment cells are recognizable, as evidenced by histochemical markers or morphology. On the other hand, morphogenesis of the notochord and other structures is dependent on precise cell movement and shape changes after the gastrula stage, which are disrupted above the high temperature threshold. These findings suggest that morphogenetic processes may be more sensitive to high temperature than cell type specification events. They also point to avenues for investigation of the limiting factors to developmental canalization in marine invertebrates.


Serum C-reactive protein concentrations in dogs with multicentric lymphoma undergoing chemotherapy.
To determine whether serum C-reactive protein (CRP) concentration is high in dogs with multicentric lymphoma, whether CRP concentration changes in response to chemotherapy, and whether CRP concentration can be used as a marker for relapse in dogs with multicentric lymphoma. Cohort study. 20 dogs with multicentric lymphoma and 8 healthy control dogs undergoing chemotherapy with cyclophosphamide, vincristine, and prednisone (CVP) or with vincristine, cyclophosphamide, methotrexate, and L-asparaginase (VCMA) and 20 other healthy dogs. Serum CRP concentration was measured weekly during the first month of chemotherapy and then at 3-week intervals until relapse in dogs with multicentric lymphoma, weekly for 16 weeks in healthy dogs undergoing chemotherapy, and once in the healthy dogs not undergoing chemotherapy. For both groups of dogs with lymphoma, mean serum CRP concentration during week 1 (prior to treatment) was significantly higher than mean concentrations following induction of chemotherapy and at the time of relapse. Mean serum CRP concentration in the healthy dogs undergoing chemotherapy was not significantly different at any time from mean concentration for the healthy dogs not undergoing chemotherapy. No significant differences were observed between dogs treated with CVP and dogs treated with VCMA. Results suggest that serum CRP concentration is high in dogs with multicentric lymphoma but that serum CRP concentration is not a useful marker for relapse and that chemotherapy itself does not affect serum CRP concentration.


The multiMiR R package and database: integration of microRNA-target interactions along with their disease and drug associations.
microRNAs (miRNAs) regulate expression by promoting degradation or repressing translation of target transcripts. miRNA target sites have been catalogued in databases based on experimental validation and computational prediction using various algorithms. Several online resources provide collections of multiple databases but need to be imported into other software, such as R, for processing, tabulation, graphing and computation. Currently available miRNA target site packages in R are limited in the number of databases, types of databases and flexibility. We present multiMiR, a new miRNA-target interaction R package and database, which includes several novel features not available in existing R packages: (i) compilation of nearly 50 million records in human and mouse from 14 different databases, more than any other collection; (ii) expansion of databases to those based on disease annotation and drug microRNAresponse, in addition to many experimental and computational databases; and (iii) user-defined cutoffs for predicted binding strength to provide the most confident selection. Case studies are reported on various biomedical applications including mouse models of alcohol consumption, studies of chronic obstructive pulmonary disease in human subjects, and human cell line models of bladder cancer metastasis. We also demonstrate how multiMiR was used to generate testable hypotheses that were pursued experimentally.


Temperature effects on the membrane current of retinal rods of the toad.
Thermal effects on the visual transduction mechanism of toad rods were examined by recording the membrane current of a single outer segment while changing the temperature within the range 15-30 degrees C. Warming increased the amplitude rmax of the saturating flash response. This effect had a Q10 of about 1.8 and may result from an increase in the light-sensitive conductance. The flash sensitivity decreased with increasing temperature, while the half-saturating flash intensity increased. There was no evidence of a temperature effect on the probability that an incident 500 nm photon triggered an electrical response. Together with the results in (2) and (3) this indicates that at higher temperature a successfully absorbed photon blocked a smaller fraction of the light-sensitive conductance. Upon warming, the time scale of the flash response shortened but the characteristic wave form was preserved. The speed of the dim flash response, measured by the reciprocal of its time-to-peak, had a Q10 of 2.7 and an apparent activation energy of 16.8 kcal mole-1. The power spectrum of the continuous component of the dark noise could be predicted at different temperatures by assuming that the underlying event was shaped by two of the four delays required to fit the light response. This behaviour is consistent with the notion that the continuous noise arises within the cascade of processes controlling the internal transmitter concentration of the outer segment.


Maackiain suppresses the development of cervical cancer via AMPK priming autophagy.
Maackiain (Mac), a flavonoid analog isolated from Sophora flavescens, exhibits neuroprotective, anti-allergic, anti-inflammatory, and pro-apoptotic effects. It is not clear whether Mac has a therapeutic effect on cervical cancer. In this work, we used RT-qPCR, western blot, immunofluorescence, and related methods to detect the therapeutic mechanism of Mac for cervical cancer. We demonstrated that Mac significantly inhibited the proliferation, migration, and invasion of human cervical cancer cell lines HeLa and SiHa. And, Mac enhanced the pro-apoptotic effects of cisplatin in treating cervical cancer cells. Mac has shown good efficacy in treating cervical cancer. Furthermore, Mac inhibited the mammalian target of the rapamycin (mTOR) pathway, thereby inducing autophagy in cervical cancer cells. The regulation of mTOR/autophagy pathway by Mac relied on the activation of AMP-activated protein kinase (AMPK), and the inhibition of the AMPK reversed the Mac's anti-cervical cancer activity. In addition, experimental study of Mac in mouse xenograft tumor model further confirmed its good anti-cervical cancer activity. Mac inhibits human cervical cancer by activating the AMPK/mTOR/autophagy pathway, indicating that it is a potential natural compound for the treatment of cervical cancer. This study also provides a feasible molecular mechanism for the treatment of cervical cancer.


Psychosocial effects of continuous oxygen therapy in hypoxaemic chronic obstructive pulmonary disease patients.
Twenty six hypoxaemic patients with severe and stable chronic obstructive pulmonary disease (COPD) were treated with continuous domiciliary oxygen for a six month period. The patients were evaluated 1, 3 and 6 months after the start of oxygen therapy. In addition to blood gas analysis, 15 coping skills were evaluated by the patient and by the nurse, who also rated the general activity of the patients. Depression was measured by Beck Depression Inventory (BDI) at the start of the trial and after six months' oxygen therapy. The general psychosocial response was meagre; no significant changes were observed in any psychosocial measures. The response was slightly better in younger and less hypoxaemic patients. We conclude that the psychosocial response to oxygen therapy in severely hypoxaemic COPD patients is limited.


Orofacial conditions and their relation to the sense of coherence among participants afflicted with leprosy in West Bengal State: A cross-sectional study.
About 4 million people are disabled by leprosy. Eighty-six percent of leprosy patients reside in Southeast Asia and Brazil. India accounts for up to 70% of total cases. In India, it is highest in West Bengal, Maharashtra, Uttar Pradesh, and Bihar. The objective of the study is to assess the quality of life in terms of sense of coherence (SOC) among patients afflicted with leprosy in the state of West Bengal (located in eastern India on the Bay of Bengal). A cross-sectional descriptive epidemiological study was conducted among estimated sample size of 350 participants who were afflicted with leprosy drawn from across the state of West Bengal, India. Data were collected on a specially designed pro forma. Most of the participants afflicted with leprosy were in the age group of 40-77 years, and there was predominance of males, with most of them belonging to the upper-lower socioeconomic class. On evaluating the incidence of extraoral lesions through ranking, facial nodules and facial spots were highest 182 (52%), while among intraoral lesions such as melanin pigmentation 176 (50%) and inflammatory papillary hyperplasia 150 (42.8%), was seen in greater fraction. The results of bivariate analysis between SOC and variables under investigation show that majority of the cases, i.e., 258 (47.14%) revealed the SOC interpretation as "poor." Statistical analysis showed significant difference in relation to age (P ≤ 0001) and socioeconomic status (P = 0.053) of participants. We conclude that 52% of the population were having facial nodules and facial spots and 50.2% of the population had intraoral manifestation such as melanin pigmentation. Analysis of SOC revealed that majority of the cases were interpreted as "poor."


Fault localization and analysis in semiconductor devices with optical-feedback infrared confocal microscopy.
We report on a cost-effective optical setup for characterizing light-emitting semiconductor devices with optical-feedback confocal infrared microscopy and optical beam-induced resistance change. We utilize the focused beam from an infrared laser diode to induce local thermal resistance changes across the surface of a biased integrated circuit (IC) sample. Variations in the multiple current paths are mapped by scanning the IC across the focused beam. The high-contrast current maps allow accurate differentiation of the functional and defective sites, or the isolation of the surface-emitting p-i-n devices in the IC. Optical beam-induced current (OBIC) is not generated since the incident beam energy is lower than the bandgap energy of the p-i-n device. Inhomogeneous current distributions in the IC become apparent without the strong OBIC background. They are located at a diffraction-limited resolution by referencing the current maps against the confocal reflectance image that is simultaneously acquired via optical-feedback detection. Our technique permits the accurate identification of metal and semiconductor sites as well as the classification of different metallic structures according to thickness, composition, or spatial inhomogeneity.


Synthesis and Biological Activity of Unnatural Enediynes.
The first reports of the natural enediyne anticancer antibiotics date back to the late 1980s; since then, a great deal of interest has been devoted to the chemistry, biology and potential medical applications of this family of compounds. The biological activity of enediynes is linked to the presence of a highly unsaturated hex-1-ene-1,5-diyne system. The thermally induced transformation of this unit into a benzene σ-σ diradical (the Bergman cycloaromatization) is the key step of the antitumor properties of such compounds: 1,4-diaryl radicals are able to abstract H-atoms from the deoxyribose backbone of DNA, thus leading to DNA strand cleavage and ultimately cell death. We undertook a structured search of bibliographic databases for peer-reviewed research literature using focused and high quality papers. Research efforts addressed at understanding and mimicking the various processes involved in the targeting, activation and DNA cleavage associated with these products are described. The potential of a great number of non natural enediynes in the treatment of many infectious diseases, apart their role in anticancer drugs, such as antibacterial activity, protein degradation activity, has been reported Results: Due to the interesting mode of action of this class of compounds, the unique molecular architecture of enediynes has been exploited towards the synthesis of many non natural compounds in order to study and enhance their biological properties. Seventy-six papers were included in this review. It is divided in paragraphs that include: Carbo- oxygen-nitrogen- and sulfur- enediynes, polymers and macrocycles. The synthetic approaches to the different classes of compounds are discussed in detail together with the biological implications of the synthesized compounds Conclusion: The review summarizes the most recent advances in the synthesis and reactivity of non natural enediynes by focusing the attention particularly to the biological properties of the most interesting members of the family of carbo- and hetero- enediynes. The findings of this review confirm the importance of non natural enediynes as potential drugs in the treatment of cancer and many infectious diseases.


Extract latent features of single-particle trajectories with historical experience learning.
Single-particle tracking has enabled real-time, in situ quantitative studies of complex systems. However, inferring dynamic state changes from noisy and undersampling trajectories encounters challenges. Here, we introduce a data-driven method for extracting features of subtrajectories with historical experience learning (Deep-SEES), where a single-particle tracking analysis pipeline based on a self-supervised architecture automatically searches for the latent space, allowing effective segmentation of the underlying states from noisy trajectories without prior knowledge on the particle dynamics. We validated our method on a variety of noisy simulated and experimental data. Our results showed that the method can faithfully capture both stable states and their dynamic switch. In highly random systems, our method outperformed commonly used unsupervised methods in inferring motion states, which is important for understanding nanoparticles interacting with living cell membranes, active enzymes, and liquid-liquid phase separation. Self-generating latent features of trajectories could potentially improve the understanding, estimation, and prediction of many complex systems.


Skeletal muscle digoxin concentration and its relation to serum digoxin concentration and cardiac effect in healthy man.
Blood samples and skeletal muscle biopsies (m. quadriceps femoris, vastus lateralis) were taken from seven healthy subjects for analysis of serum and skeletal muscle digoxin concentrations by radioimmunoassay using a percutaneous needle biopsy technique for muscle sampling. The subjects were investigated on two digoxin dose levels and on the third day after withdrawal of digoxin. It was found that the skeletal muscle/serum digoxin ratio was significantly higher than the corresponding ratio obtained in a previous study with muscle sampling (m. rectus abdominis) from patients during open heart surgery. The present study indicates a significant correlation between the digoxin concentrations in serum and skeletal muscle as well as between cardiac effect, measured by changes in QS2I, and skeletal muscle digoxin concentration. A doubling of the digoxin dose gave a proportional increase in skeletal muscle digoxin concentration. The magnitude of the estimated half-life of skeletal muscle digoxin was the same as previously reported for serum or plasma digoxin.


[Experience of local radiation therapy and total irradiation of the skin by electron beam in patients with primary B and T-cell lymphomas of the skin].
The problem of the treatment of primary malignant lymphomas of the skin is now becoming increasingly important due to the increase of cases among people of working age and disability of these patients. In most cases lymphomas of the skin have a T-cell origin, the most common of skin lymphoma is mycosis fungoides. It is poorly studied the role of electronic radiation therapy in local and systemic skin lymphomas as well as methodological questions of its application, so research in this field is actual. Therefore the aim of the study is improving of the efficiency of therapy in patients affected by non-Hodgkin's lymphoma with skin lesions by the use of local radiation therapy and total skin irradiation by electron beam.


Ulcerative enterocolitis in two goats associated with enterotoxin- and beta2 toxin-positive Clostridium perfringens type D.
Enterotoxemia caused by Clostridium perfringens type D in sheep is believed to result from the action of epsilon toxin (ETX). However, the sole role of ETX in the intestinal changes of the acute and chronic forms of enterotoxemia in goats remains controversial, and the synergistic action of other C. perfringens toxins has been suggested previously. The current study examined 2 goats that were found dead without premonitory clinical signs. Gross lesions at necropsy consisted of multifocal fibrinonecrotic enterocolitis, edematous lungs, and excess pleural fluid. Histologically, there were multifocal fibrinonecrotic and ulcerative ileitis and colitis, edema of the colonic serosa, and proteinaceous interstitial edema of the lungs. Clostridium perfringens type D carrying the genes for enterotoxin (CPE) and beta2 toxin (CPB2) was cultured from intestinal content and feces of 1 of 2 goats, while C. perfringens type D CPB2-positive was isolated from the other animal. When multiple colonies of the primary isolations from both animals were tested by Western blot, most of the isolates expressed CPB2, and only a few isolates from the first case expressed CPE. Alpha toxin and ETX were detected in ileal and colonic contents and feces of both animals by antigen capture enzyme-linked immunosorbent assay. CPB2, but not CPE, was identified in the small and large intestines of both goats by immunohistochemistry. These findings indicate that CPB2 may have contributed to the necrotic changes observed in the intestine, possibly assisting ETX transit across the intestinal mucosa.


Impact of a Combination of UV-C Irradiation and Peracetic Acid Spray Treatment on Brochothrix thermosphacta and Yersinia enterocolitica Contaminated Pork.
Efficient ways of decontamination are needed to minimize the risk of infections with Yersinia (Y.) enterocolitica, which causes gastrointestinal diseases in humans, and to reduce the numbers of Brochothrix (B.) thermosphacta to extend the shelf-life of meat. While many studies have focused on a single treatment of peracetic acid (PAA) or UV-C-irradiation, there are no studies about a combined treatment on meat. Therefore, in the present study, pork was inoculated with either Y. enterocolitica or B. thermosphacta, and was treated with a combination of 2040 mJ/cm2 UV-C irradiation followed by a 2000 ppm PAA spray treatment (30 s). Samples were packed under modified atmosphere and stored for 1, 7, or 14 days. The samples were examined for Y. enterocolitica and B. thermosphacta content, chemical and sensory effects, and meat quality parameters. For Y. enterocolitica, a significant reduction of up to 2.16 log10 cfu/cm2 meat and for B. thermosphacta, up to 2.37 log10 cfu/cm2 meat was seen on day 14 after UV-C/PAA treatment compared to the untreated controls.


Selective facilitation of responses to cortical stimulation of proximal and distal arm muscles by precision tasks in man.
1. The responses of the first dorsal interosseus (1DI), opponens pollicis (OP), extensor digitorum communis (EDC), brachioradialis (BR), biceps brachii (BB) and anterior deltoid (AD) muscles to magnetic stimulation of the motor cortex were recorded during different motor tasks. 2. Two precision and two power isometric tasks were investigated. The precision tasks were a pincer grip ('grip') and a thrust against a target with the wrist ('push'). In the former, the prime movers were the intrinsic hand muscles, while the proximal muscles played a postural role. In the latter, the prime movers were the proximal muscles. In both tasks, force was controlled through visual feedback. The power tasks required encirclement of a cylinder with the fingers ('grasp'), or sustaining a weight suspended at wrist level ('load'). 3. Magnetic stimulation was applied in eight subjects by a coil placed over the vertex at 1.1-1.2 times the motor threshold for the most excitable muscles. This produced in the prime mover muscles larger motor-evoked responses (MEPs) during grip or push tasks than grasp or load tasks, in spite of similar background EMG levels. During grip tasks, only one of the two prime movers showed task-dependent changes. In the postural muscle AD there was no significant difference between MEPs during grip and grasp tasks; however, BB responses were larger during grasp than grip tasks. 4. MEPs simultaneously recorded in the prime movers were plotted against each other. The slope of the regression line for AD versus BB was larger in push than load tasks, whilst the changes in MEPs of 1DI and OP were independent during both grip and grasp tasks. 5. In three subjects, MEPs were also elicited by electrical stimulation during grip and grasp tasks. MEP changes tended to parallel those obtained for magnetic stimulation, but the increase in size of the electrically evoked MEPs during the precision task was smaller. 6. In all subjects the median and ulnar nerves were stimulated during grip and grasp tasks, and an H reflex was evoked in the hand muscles of five subjects. In no case did the two tasks produce reflexes of different amplitude. 7. The motor response of both proximal and distal muscles can be task dependent, in spite of the differences in their principal functional role and cortical representation. The modulation is related to the degree of control requested by the task, and is likely to reflect selective changes in the excitability of corticospinal neurones.


Filamentous sulfur bacteria preserved in modern and ancient phosphatic sediments: implications for the role of oxygen and bacteria in phosphogenesis.
Marine phosphate-rich sedimentary deposits (phosphorites) are important geological reservoirs for the biologically essential nutrient phosphorous. Phosphorites first appear in abundance approximately 600 million years ago, but their proliferation at that time is poorly understood. Recent marine phosphorites spatially correlate with the habitats of vacuolated sulfide-oxidizing bacteria that store polyphosphates under oxic conditions to be utilized under sulfidic conditions. Hydrolysis of the stored polyphosphate results in the rapid precipitation of the phosphate-rich mineral apatite-providing a mechanism to explain the association between modern phosphorites and these bacteria. Whether sulfur bacteria were important to the formation of ancient phosphorites has been unresolved. Here, we present the remains of modern sulfide-oxidizing bacteria that are partially encrusted in apatite, providing evidence that bacterially mediated phosphogenesis can rapidly permineralize sulfide-oxidizing bacteria and perhaps other types of organic remains. We also describe filamentous microfossils that resemble modern sulfide-oxidizing bacteria from two major phosphogenic episodes in the geologic record. These microfossils contain sulfur-rich inclusions that may represent relict sulfur globules, a diagnostic feature of modern sulfide-oxidizing bacteria. These findings suggest that sulfur bacteria, which are known to mediate the precipitation of apatite in modern sediments, were also present in certain phosphogenic settings for at least the last 600 million years. If polyphosphate-utilizing sulfide-oxidizing bacteria also played a role in the formation of ancient phosphorites, their requirements for oxygen, or oxygen-requiring metabolites such as nitrate, might explain the temporal correlation between the first appearance of globally distributed marine phosphorites and increasing oxygenation of Neoproterozoic oceans.


[Nurse-patient relations in a psychiatric department (experience with a social psychology study)].
The paper is related to some aspects of "nurse-patient" interrelations in conditions of rehabilitation of mental patients. The authors studied some factors influencing the evaluation (selection) of nurses by patients with the aid of special questionnaire techniques of nurses and patients in combination with the method of expert assessment. The study covered 141 mental patients and 14 nurses working in shifts. It was possible to distinguish some personality and occupational characteristics of the nurses who were most closely related to the estimation made by the patients. The authors established some relation between the assessment made by the patients with the degree of readiness for social contacts with mental patients by nurses.


Endoplasmic reticulum protein ERp46 in prostate adenocarcinoma.
Endoplasmic reticulum (ER) protein ERp46 is a member of the protein disulfide isomerase family of oxidoreductases, which facilitates the reduction of disulfides in proteins and their folding. Accumulation of misfolded proteins has been implicated in cancer. The objectives of the present study were to investigate the role of ERp46 in prostate cancer, its expression and its effects on prostate cancer growth. A tissue microarray with human prostate cancer and normal prostate tissue samples was stained for ERp46 followed by image analysis. Human prostate adenocarcinoma 22Rv1 cells were stably transfected with short hairpin RNA (shRNA) specific for ERp46, a non-effective scrambled control or a plasmid containing full-length human ERp46 cDNA, and cell growth was determined. Subcloned cells were treated with thapsigargin or tunicamycin to induce ER stress and lysates were subjected to western blot analysis for ER stress proteins. Subcutaneous xenografts of parental 22Rv1, ERp46-overexpressing (ERp46+), shERp46 or scrambled control cells were established in male inbred BALB/c nude mice (n=10/group). Tumor growth curves of the xenografts were constructed over a period of 30 days and subsequently the mice were sacrificed and the amount of serum prostate-specific antigen was determined. The results demonstrated increased ERp46 expression levels in prostate cancer tissue samples of Gleason ≥7 compared with normal prostate tissue samples. When ERp46 was stably knocked down using shRNA or overexpressed in prostate carcinoma 22Rv1 cells, tumor growth in vitro and in BALB/c nude mice was inhibited and accelerated, respectively. ERp46 overexpression led to reduced sensitivity to ER stress as indicated by higher half maximal inhibitory concentrations for tunicamycin and thapsigargin in ERp46+ cells. The shERp46 cells lost the ability to upregulate protein disulfide isomerase following tunicamycin-induced ER stress. The present study suggests a role for ERp46 as a therapeutic target in prostate cancer, given its expression profile in human prostate cancer, and its effect on prostate cancer cell growth.


Endoscopic Ultrasound-Guided Biliary Drainage for Unresectable Hilar Malignant Biliary Obstruction.
Endoscopic transpapillary biliary drainage is the current standard of care for unresectable hilar malignant biliary obstruction (MBO) and bilateral metal stent placement is shown to have longer patency. However, technical and clinical failure is possible and percutaneous transhepatic biliary drainage (PTBD) is sometimes necessary. Endoscopic ultrasound-guided biliary drainage (EUS-BD) is increasingly being reported as an alternative rescue procedure to PTBD. EUS-BD has a potential advantage of not traversing the biliary stricture and internal drainage can be completed in a single session. Some approaches to bilateral biliary drainage for hilar MBO under EUS-guidance include a bridging method, hepaticoduodenostomy, and a combination of EUS-BD and transpapillary biliary drainage. The aim of this review is to summarize data on EUS-BD for hilar MBO and to clarify its advantages over the conventional approaches such as endoscopic transpapillary biliary drainage and PTBD.


Total flavones from Sonchus arvensis L. ameliorate colitis by adjusting the gut microbiota.
Sonchus arvensis L. is traditional Chinese food and medicine. We investigated protective effects of flavones from Sonchus arvensis L. (SAF) on colitis induced by dextran sulfate sodium (DSS) in mice by regulating gut microbiota (GM). C57BL/6 mice were divided randomly: control group (CL); DSS group (ML); positive control + DSS group (AN); SAF + DSS (FE) group. The protective effects of SAF on ulcerative colitis (UC) were estimated by food intake, water intake, bodyweight loss, diarrhea, blood in stools, colon length, histology, disease activity index (DAI) score, and blood parameters. The sequencing of 16S rRNA gene was detected to investigate effect of SAF on GM. SAF attenuate bodyweight loss significantly. The DAI score was lower in FE group than that in ML group. Colon length was improved significantly in ML group. Pathologic changes could be ameliorated after SAF was administered to UC mice. SAF improved blood parameters of model mice. 16S rRNA sequencing revealed that it was very important to ameliorate colitis with bacteria of the phylum Verrucomicrobiota, class Verrucomicrobiae, order Verrucomicrobiales, family Akkermansiaceae, and genus Akkermansia. The SAF protective effect against colitis induced by DSS in mice may have a connection with GM diversity.


Evaluation of pancreatic endocrine and exocrine function in patients with autoimmune pancreatitis.
Up to now, the characteristics of pancreatic endocrine and exocrine functions in autoimmune pancreatitis (AIP) are still unclear. The aim of this study is to evaluate pancreatic functions in AIP compared with those of chronic pancreatitis (CP). Twelve patients with AIP and 25 patients with CP were examined for exocrine and endocrine pancreas. Exocrine function was evaluated by a secretin test. Concerning endocrine function, insulin secretion (C-peptide response) was examined with the glucagon tolerance test and glucagon secretion was examined with the arginine tolerance test. Pathological examination of pancreatic tissues was done on the operative specimens of AIP and CP that could not be clinically excluded from pancreatic cancer. For the secretin test, 8.3% of patients with AIP showed 1-factor abnormality, which was a reduction in volume, and 41.7% showed 2-factor abnormalities, which were a reduction in volume and amylase output. On the other hand, 44.0% of patients with CP showed only 1-factor abnormality, which was the reduction in the maximum bicarbonate concentration. Autoimmune pancreatitis accompanied with diabetes mellitus showed a reduction both in DeltaC-peptide response (beta-cell response) and Deltaglucagon (alpha-cell response). Histologically, AIP showed lymphoplasmatic cells infiltration surrounding the pancreatic ducts, but basement membranes were intact. Moreover, basement membranes of the duct were injured in CP. Furthermore, islet cells in AIP were revealed as almost intact even though they were surrounded by fibrosis. These findings indicate that exocrine dysfunction with AIP is different from CP because AIP induces stenosis of the pancreatic ducts, but ductal cells that possess the function of bicarbonate secretion are intact, and that endocrine dysfunction with AIP was secondary pancreatic diabetes.


Macrocognition in Day-To-Day Police Incident Response.
Using examples of incidents that UK Police Forces deal with on a day-to-day basis, we explore the macrocognition of incident response. Central to our analysis is the idea that information relating to an incident is translated from negotiated to structured and actionable meaning, in terms of the Community of Practice of the personnel involved in incident response. Through participant observation of, and interviews with, police personnel, we explore the manner in which these different types of meaning shift over the course of incident. In this way, macrocognition relates to gathering, framing, and sharing information through the collaborative sensemaking practices of those involved. This involves two cycles of macrocognition, which we see as 'informal' (driven by information gathering as the Community of Practice negotiates and actions meaning) and 'formal' (driven by the need to assign resources to the response and the need to record incident details). The examples illustrate that these cycles are often intertwined, as are the different forms of meaning, in situation-specific ways that provide adaptive response to the demands of the incident.


[Changes in the monoamine content in different parts of hypothalamus depending on the stages of the estrous cycle].
Fluorimetric determination of monoamines in various regions of the hypothalamus and at different stages of the estral cycle in rats showed that the serotonin, noradrenaline, and particularly dophamine content changed both in the course of the cycle and at different time (10, 15 and 18 hours) of the same stage of the cycle. Dophamine concentration in the arcuate area--the centre of the tonic activity--reached its maximum at 18 hours of the diestrus-2 (D2) and fell to the minimum at 10 hours of the proestrus (P). Noradrenaline level in the preoptic area increased at 18 hours of the D2 and fell at 10 hours of the P. It is supposed that in the hypothalamic regulation of the estral cycle at least two monoamines (dopamine and noradrenaline) took part; the trigger role belongs to noradrenaline of the preoptic area (the cyclic centre).


PBT2 Reduces Toxicity in a C. elegans Model of polyQ Aggregation and Extends Lifespan, Reduces Striatal Atrophy and Improves Motor Performance in the R6/2 Mouse Model of Huntington's Disease.
There is evidence that interaction with biologically important metals, particularly copper and iron, contributes to the pathological aggregation and toxicity of the mutant huntingtin protein in HD. PBT2 is a novel 8-hydroxyquinoline drug in clinical trials for AD and HD which restores metal homeostasis and has demonstrated neuroprotective and cognitive benefits in animal models of AD and in a Phase IIa clinical trial in AD patients. We assessed efficacy of PBT2 to improve function in nematode and mouse models of HD. We assessed the effects of PBT2 on motility in a strain of C. elegans engineered to overexpress an extended polyglutamine tract. We then assessed the effects of daily oral administration of PBT2 on the R6/2 mouse a vertebrate model of HD. Behavioural measures (Rotarod performance, clasping latency and duration) and measures of overall health (body weight, lifespan) were assessed throughout the study. At conclusion brains were evaluated for weight and lateral ventricle area. PBT2 significantly decreased paralysis caused by overexpression of an extended polyglutamine tract in a C. elegans. R6/2 mice treated with PBT2 showed significantly better performance in the Rotarod task and significantly decreased duration of clasping than control animals. Mean body weight and brain weight in the PBT2 treatment group were significantly higher than the control cohort and median lifespan was prolonged by 26%. Mean lateral ventricle area in the PBT2-treated group was 46% smaller than in the control group. Treatment with PBT2 may be beneficial as a disease modifying therapy for HD.


Dioxin-dependent activation of murine Cyp1a-1 gene transcription requires protein kinase C-dependent phosphorylation.
Transcriptional activation of the murine Cyp1a-1 (cytochrome P(1)450) gene by inducers such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (dioxin) requires the aromatic hydrocarbon (Ah) receptor and the interaction of an inducer-receptor complex with one or more of the Ah-responsive elements (AhREs) located about 1 kb upstream from the transcriptional initiation site. We find that treatment of mouse hepatoma Hepa-1 cells with 2-aminopurine, an inhibitor of protein kinase activity, inhibits CYP1A1 mRNA induction by TCDD as well as the concomitant increase in CYP1A1 enzyme activity. Formation of DNA-protein complexes between the Ah receptor and its AhRE target is also inhibited by 2-aminopurine, as determined by gel mobility shift assays. Phosphorylation is required for the formation of Ah receptor-specific complexes, since in vitro dephosphorylation of nuclear extracts from TCDD-treated Hepa-1 cells abolishes the capacity of the Ah receptor to form specific complexes with its cognate AhRE sequences. To determine whether any one of several known protein kinases was involved in the transcriptional regulation of the Cyp1a-1 gene, we treated Hepa-1 cells with nine other protein kinase inhibitors prior to induction with TCDD; nuclear extracts from these cells were analyzed for their capacity to form specific DNA-protein complexes. Only extracts from cells treated with staurosporine, a protein kinase C inhibitor, were unable to form these complexes. In addition, staurosporine completely inhibited CYP1A1 mRNA induction by TCDD. Depletion of protein kinase C by prolonged treatment with phorbol ester led to the complete suppression of CYP1A1 mRNA induction by TCDD. We conclude that (i) phosphorylation is necessary for the formation of a transcriptional complex and for transcriptional activation of the Cyp1a-1 gene; (ii) the phosphorylation site(s) exists on at least one of the proteins constituting the transcriptional complex, possibly the Ah receptor itself; and (iii) the enzyme responsible for the phosphorylation is likely to be protein kinase C.


Serum IgE concentrations in neurological diseases.
Serum IgE levels were determined in 42 patients with Guillain Barre Syndrome and 22 patients with multiple sclerosis. Serum IgE concentrations were significantly elevated in about half of the patients with acute Guillain Barre Syndrome and tends to fall in patients with clinical improvement. Patients with multiple sclerosis and other acute and chronic central nervous system and peripheral system diseases have normal serum IgE levels. IgE is an important mediator of tissue damage in allergic diseases of immediate type and may also play an important role in acute Guillain Barre Syndrome.