Estimates of the Economic Cost Caused by Five Major Reproductive Problems in Dairy Animals in Assam and Bihar, India.
Reproductive problems in dairy animals reduce fertility, prevent conception, create problems in the delivery of healthy calves, lead to postpartum complications, increase inter-calving periods, reduce milk yield, and lower overall lifetime productivity. This study aimed at understanding the incidence of reproductive problems and the cost caused by these. The study covered 954 dairy animals in Bihar and 1348 dairy animals in Assam that were selected using a multi-stage random sampling method. The costs were calculated as the sum of income losses and expenditures incurred. The major cost incurred resulted from extended calving intervals (46.1% of the total cost), followed by loss through salvage selling (38.1%), expenditure for treatment of repeat breeders (5.9%), loss of milk production (5.3%) and expenditure for extra inseminations (2.0%). About one fifth of the selected reproductive problems were left untreated. The estimated cost of reproductive problems was Indian Rupees (INR) 2424.9 (USD 36.1) per dairy animal per year (of the total dairy animal population) which represented approximately 4.1% of the mean value loss of dairy animals (INR 58,966/USD 877) per year. Reproductive problems were significantly (p < 0.001) higher among improved (exotic breed or cross-bred) dairy animals than indigenous (native breed or nondescript indigenous) dairy animals. The study suggests that with the increase of improved dairy animal population, the loss may further increase. The study concludes that any economic estimation of reproduction problems based on aetiology without confirmatory diagnoses could be highly misleading because of the complex nature of the problems.


Natural Infection of Pomegranate (Punica Granatum) by Apple Dimple Fruit Viroid.
The analysis by high throughput sequencing (HTS) and RT-PCR of Spanish pomegranate fruits showing yellow rings revealed the presence of viroid isolates closely related to fig isolates of apple dimple fruit viroid (ADFVd). The analysis of pomegranate public RNASeq data (Sequence Reads Archives, SRAs) from Israel provided evidence for the presence of similar ADFVd isolates in pomegranate trees in this country. In addition, reads or contigs of plum viroid I (PVd-I) isolates were also identified in two of the analyzed SRA datasets from Israel, suggesting the presence of this second viroid in pomegranate. Full length ADFVd genomic sequences have been recovered, increasing knowledge on the diversity of this viroid and on the pomegranate virome in which only four viruses and one viroid had previously been reported.


[Evaluation of the intensity of stress at work and burnout syndrome in the managers].
Burn out syndrome is connected with the intensity of stress at work. It is a common problem among people who work in management departments. This significantly decreased the effectiveness of work and the quality of psychosocial functioning. The purpose of the study was to establish the dependence between occupational stress and burnout syndrome in the group of managers. The studied group consisted of 73 managers, aged 27-55 years. The Perceived Job Stress Questionnaire and Maslach Burnout Inventory (MBI) were used in this study. The data obtained revealed significant intensity of stress filling at work in evaluated group. The most stressful factors occur: psychical balance of work, lack of rewards, social contacts and uncertainty at work. The highest risk of burnout syndrome is associated with symptoms of emotional exhaustion and depersonalization. Higher intensity of occupational stress is connected with the greater intensification of burnout syndrome in investigated group of managers.


Physicians' shared decision-making behaviors in depression care.
Although shared decision making (SDM) has been reported to facilitate quality care, few studies have explored the extent to which SDM is implemented in primary care and factors that influence its application. This study assesses the extent to which physicians enact SDM behaviors and describes factors associated with physicians' SDM behaviors within the context of depression care. In a secondary analysis of data from a randomized experiment, we coded 287 audiorecorded interactions between physicians and standardized patients (SPs) using the Observing Patient Involvement (OPTION) system to assess physician SDM behaviors. We performed a series of generalized linear mixed model analyses to examine physician and patient characteristics associated with SDM behavior. The mean (SD) OPTION score was 11.4 (3.3) of 48 possible points. Older physicians (partial correlation coefficient = -0.29; beta = -0.09; P < .01) and physicians who practiced in a health maintenance organization setting (beta = -1.60; P < .01) performed fewer SDM behaviors. Longer visit duration was associated with more SDM behaviors (partial correlation coefficient = 0.31; beta = 0.08; P < .01). In addition, physicians enacted more SDM behaviors with SPs who made general (beta = 2.46; P < .01) and brand-specific (beta = 2.21; P < .01) medication requests compared with those who made no request. In the context of new visits for depressive symptoms, primary care physicians performed few SDM behaviors. However, physician SDM behaviors are influenced by practice setting and patient-initiated requests for medication. Additional research is needed to identify interventions that encourage SDM when indicated.


Dairy products and the risk of developing prostate cancer: A large-scale cohort study (JACC Study) in Japan.
Dairy products have been indicated as a risk factor for prostate cancer. However, only a few epidemiological studies have reported dairy products as being a risk factor for prostate cancer in Japan, reporting contradictory results. We therefore investigated the association between the intake of dairy products and the occurrence of prostate cancer through a large-scale cohort study. The Japan Collaborative Cohort study analyzed approximately 110,000 residents from various Japanese districts who participated in our questionnaire survey during 1988-1990. The subjects of the present study were 26,464 men (age range: 40-79 years) from 24 districts wherein cancer incidence was reported. Their clinical course was followed up until 2009. Hazard ratios (HRs) were calculated using Cox's proportional hazards model, adjusted for age, survey area, family history of prostate cancer, body mass index, and total energy intake. For diet, we calculated the HRs associated with intermediate and high consumption of dairy products and compared them with those associated with low consumption. There were 412 cases of prostate cancer in the survey population. As dairy products, milk, yogurt, cheese, and butter were evaluated. Among them, milk consumption was associated with a significant risk (HR = 1.37, p = 0.009) and a dose-dependent response (p for trend = 0.009) adjusted for age and family history of prostate cancer, stratified by area. Milk and yogurt consumption showed a significantly positive risk and a dose-response relationship adjusted for age, family history of prostate cancer, body mass index, and total energy intake, stratified by area. In summary, a high intake of dairy products such as milk increased the risk of developing prostate cancer in Japanese men.


Time for a change to assess and evaluate body temperature in clinical practice.
The definition of normal body temperature as 37 degrees C still is considered the norm worldwide, but in practice there is a widespread confusion of the evaluation of body temperature, especially in elderly individuals. In this paper, we discuss the relevance of normal body temperature as 37 degrees C and consequences in clinical practice. Our conclusion is that body temperature should be evaluated in relation to the individual variability and that the best approach is to use the same site, and an unadjusted mode without adjustments to other sites. If the baseline value is not known, it is important to notice that frail elderly individuals are at risk of a low body temperature. In addition, what should be regarded as fever is closely related to what is considered as normal body temperature. That is, as normal body temperature shows individual variations, it is reasonable that the same should hold true for the febrile range.


Adenovirus core protein V interacts with p32--a protein which is associated with both the mitochondria and the nucleus.
Adenovirus protein V is associated with the DNA-containing virus core and functions as a bridge between the capsid and the core. A yeast two-hybrid analysis performed with a human cDNA library using protein V as 'bait' selected a cellular protein, p32 --described previously as associated with the splicing factor ASF/SF2. By expression and purification of p32 and preparation of an antibody we confirmed the binding of p32 to V by a variety of methods including immune precipitation. We demonstrated that p32 was primarily located in the cytoplasm in association with mitochondria but could also be detected in the nucleus as distinct granules and tubules. By examining infected cells using confocal microscopy and immunofluorescence we were able to follow the intracellular locations of protein V and p32 and it is postulated that p32 is part of a system which imports proteins to the nucleus and that adenovirus hijacks this process to deliver its genome to the nucleus.


Reference values for volumetric capnography-derived non-invasive parameters in healthy individuals.
The aim of this study was to determine typical values for non-invasive volumetric capnography (VCap) parameters for healthy volunteers and anesthetized individuals. VCap was obtained by a capnograph connected to the airway opening. We prospectively studied 33 healthy volunteers 32 ± 6 years of age weighing 70 ± 13 kg at a height of 171 ± 11 cm in the supine position. Data from these volunteers were compared with a cohort of similar healthy anesthetized patients ventilated with the following settings: tidal volume (VT) of 6-8 mL/kg, respiratory rate 10-15 bpm, PEEP of 5-6 cmH₂O and FiO₂ of 0.5. Volunteers showed better clearance of CO₂ compared to anesthetized patients as indicated by (median and interquartile range): (1) an increased elimination of CO₂ per mL of VT of 0.028 (0.005) in volunteers versus 0.023 (0.003) in anesthetized patients, p < 0.05; (2) a lower normalized slope of phase III of 0.26 (0.17) in volunteers versus 0.39 (0.38) in anesthetized patients, p < 0.05; and (3) a lower Bohr dead space ratio of 0.23 (0.05) in volunteers versus 0.28 (0.05) in anesthetized patients, p < 0.05. This study presents reference values for non-invasive volumetric capnography-derived parameters in healthy individuals. Mechanical ventilation and anesthesia altered these values significantly.


Impact of polygenic risk communication: an observational mobile application-based coronary artery disease study.
We developed a smartphone application, MyGeneRank, to conduct a prospective observational cohort study (NCT03277365) involving the automated generation, communication, and electronic capture of response to a polygenic risk score (PRS) for coronary artery disease (CAD). Adults with a smartphone and an existing 23andMe genetic profiling self-referred to the study. We evaluated self-reported actions taken in response to personal CAD PRS information, with special interest in the initiation of lipid-lowering therapy. 19% (721/3,800) of participants provided complete responses for baseline and follow-up use of lipid-lowering therapy. 20% (n = 19/95) of high CAD PRS vs 7.9% (n = 8/101) of low CAD PRS participants initiated lipid-lowering therapy at follow-up (p-value = 0.002). Both the initiation of statin and non-statin lipid-lowering therapy was associated with degree of CAD PRS: 15.2% (n = 14/92) vs 6.0% (n = 6/100) for statins (p-value = 0.018) and 6.8% (n = 8/118) vs 1.6% (n = 2/123) for non-statins (p-value = 0.022) in high vs low CAD PRS, respectively. High CAD PRS was also associated with earlier initiation of lipid lowering therapy (average age of 52 vs 65 years in high vs low CAD PRS respectively, p-value = 0.007). Overall, degree of CAD PRS was associated with use of any lipid-lowering therapy at follow-up: 42.4% (n = 56/132) vs 28.5% (n = 37/130) (p-value = 0.009). We find that digital communication of personal CAD PRS information is associated with increased and earlier lipid-lowering initiation in individuals of high CAD PRS. Loss to follow-up is the primary limitation of this study. Alternative communication routes, and long-term studies with EHR-based outcomes are needed to understand the generalizability and durability of this finding.


Antibodies against Anthrax Toxins: A Long Way from Benchlab to the Bedside.
Anthrax is an acute disease caused by the bacterium Bacillus anthracis, and is a potential biowarfare/bioterrorist agent. Its pulmonary form, caused by inhalation of the spores, is highly lethal and is mainly related to injury caused by the toxins secretion. Antibodies neutralizing the toxins of B. anthracis are regarded as promising therapeutic drugs, and two are already approved by the Federal Drug Administration. We developed a recombinant human-like humanized antibody, 35PA83 6.20, that binds the protective antigen and that neutralized anthrax toxins in-vivo in White New Zealand rabbits infected with the lethal 9602 strain by intranasal route. Considering these promising results, the preclinical and clinical phase one development was funded and a program was started. Unfortunately, after 5 years, the preclinical development was cancelled due to industrial and scientific issues. This shutdown underlined the difficulty particularly, but not only, for an academic laboratory to proceed to clinical development, despite the drug candidate being promising. Here, we review our strategy and some preliminary results, and we discuss the issues that led to the no-go decision of the pre-clinical development of 35PA83 6.20 mAb. Our review provides general information to the laboratories planning a (pre-)clinical development.


Morphology control of structured polymer brushes.
The surface-initiated photopolymerization (SIPP) of vinyl monomers on structured self-assembled monolayers, as defined by two-dimensional (2D) initiator templates for polymer growth, is investigated. The 2D templates are prepared by electron-beam chemical lithography (EBCL) of 4'-nitro-4-mercaptobiphenyl (NBT) and chemical conversion to an asymmetric azo initiator (4'-azomethylmalonodinitrile-1,1'-biphenyl-4-thiol). Ex situ kinetic studies of the SIPP process reveal a linear increase in the thickness of the polymer layer with the irradiation/polymerization time. The effect of the applied electron dosage during the EBCL process upon the final thickness of the polymer layer is also studied. The correlation between the electron-induced conversion of the 4'-nitro to the 4'-amino group and the layer thickness of the resulting polymer brush indicates that the polymer-brush grafting density can be directly controlled by the EBCL process. NBT-based template arrays are used for the combinatorial study of the influence of the lateral structure size and the irradiation dosage on the morphology of the resulting polymer-brush layer. Analysis of the array topography reveals the dependence of the thickness of the dry polymer layer on both electron dosage and structure size. This unique combination of EBCL as a lithographic technique to locally manipulate the surface chemistry and SIPP to amplify the created differences allows the preparation of defined polymer-brush layers of controlled morphologies.


Context-specific metabolic networks are consistent with experiments.
Reconstructions of cellular metabolism are publicly available for a variety of different microorganisms and some mammalian genomes. To date, these reconstructions are "genome-scale" and strive to include all reactions implied by the genome annotation, as well as those with direct experimental evidence. Clearly, many of the reactions in a genome-scale reconstruction will not be active under particular conditions or in a particular cell type. Methods to tailor these comprehensive genome-scale reconstructions into context-specific networks will aid predictive in silico modeling for a particular situation. We present a method called Gene Inactivity Moderated by Metabolism and Expression (GIMME) to achieve this goal. The GIMME algorithm uses quantitative gene expression data and one or more presupposed metabolic objectives to produce the context-specific reconstruction that is most consistent with the available data. Furthermore, the algorithm provides a quantitative inconsistency score indicating how consistent a set of gene expression data is with a particular metabolic objective. We show that this algorithm produces results consistent with biological experiments and intuition for adaptive evolution of bacteria, rational design of metabolic engineering strains, and human skeletal muscle cells. This work represents progress towards producing constraint-based models of metabolism that are specific to the conditions where the expression profiling data is available.


Maximizing the Smile Symmetry in Facial Paralysis Reconstruction: An Algorithm Based on Twenty Years' Experience.
Over the last two decades, the senior author (B.H.) has had an extensive experience with facial paralysis reconstruction. During this period, the techniques have evolved substantially based on the experience and after observing and analyzing the surgical outcomes. The purpose of this article is to relay the lessons learned from the 20 years' experience and suggest an algorithm. In this retrospective study, we have included 343 cases of facial paralysis cases. Complete facial paralysis cases were 285 and 58 were incomplete facial paralyses, both requiring surgical procedures. Complete facial paralyses were divided in to short term (n = 83) and long term (n = 202). In total, 58% of the patients were women and 42% were men. The age range was 6 to 82 years. The techniques employed were direct suture, nerve grafts, cross-facial nerve grafts (CFNGs), masseteric-to-facial nerve transference, hypoglossal-to-facial nerve transference, free muscle transplants, and lengthening temporal myoplasty to achieve the best symmetry after reanimation of unilateral, bilateral, complete, and incomplete facial paralysis. The type of paralysis, objective measurements, the personal patient's smile, and the gender are key concepts to be considered before scheduling a dynamic facial paralysis reconstruction. For unilateral facial paralysis, the time of onset, the type of paralysis, the patient's comorbidities, and the healthy side status are some of the determining factors when selecting the correct technique. The preferred techniques for unilateral facial paralysis are direct repair, CFNG, masseteric-to-facial transposition, and free gracilis transfer. For incomplete facial paralysis, the masseteric-to-facial nerve transference is preferred. In bilateral facial paralysis, bilateral free gracilis transfer is performed in two stages using the nerve of the masseter muscle as the source of innervation. The authors provide an algorithm which simplifies facial paralysis reconstruction to achieve the greatest facial symmetry while thinking about the potential comorbidities and developing spontaneity smile according to the gender of the patient.


4β-Hydroxycholesterol is a prolipogenic factor that promotes SREBP1c expression and activity through the liver X receptor.
Oxysterols are oxidized derivatives of cholesterol that play regulatory roles in lipid biosynthesis and homeostasis. How oxysterol signaling coordinates different lipid classes such as sterols and triglycerides remains incompletely understood. Here, we show that 4β-hydroxycholesterol (HC) (4β-HC), a liver and serum abundant oxysterol of poorly defined functions, is a potent and selective inducer of the master lipogenic transcription factor, SREBP1c, but not the related steroidogenic transcription factor SREBP2. By correlating tracing of lipid synthesis with lipogenic gene expression profiling, we found that 4β-HC acts as a putative agonist for the liver X receptor (LXR), a sterol sensor and transcriptional regulator previously linked to SREBP1c activation. Unique among the oxysterol agonists of the LXR, 4β-HC induced expression of the lipogenic program downstream of SREBP1c and triggered de novo lipogenesis both in primary hepatocytes and in the mouse liver. In addition, 4β-HC acted in parallel to insulin-PI3K-dependent signaling to stimulate triglyceride synthesis and lipid-droplet accumulation. Thus, 4β-HC is an endogenous regulator of de novo lipogenesis through the LXR-SREBP1c axis.


6-Thioguanine-induced DNA damage as a determinant of cytotoxicity in cultured Chinese hamster ovary cells.
The mechanism of action of 6-thioguanine (TG) has been examined in cultured Chinese hamster ovary cells by direct measurement of the incorporation of the compound into DNA and by analysis of the resulting DNA damage. The predominant lesions as monitored by alkaline elution were DNA strand breaks. Very few, if any, interstrand or DNA-protein cross-links could be definitively observed. The cytotoxicity of TG as measured by colony-forming ability appeared closely related with its incorporation into DNA and the DNA strand scission events. As TG concentrations were increased, cytotoxicity, DNA incorporation, and strand scission reached a plateau; this result is consistent with earlier reports that TG produces a reversible block of DNA synthesis. Strand breaks appeared to be related to the incorporation of TG into DNA, since the addition of 1 microM cycloheximide during a 24-hr treatment with 3 microM TG prevented the cytotoxicity, prevented incorporation of TG into DNA, and eliminated the strand breaks. Alkali-labile sites were detected in the DNA of TG-treated cells by alkaline elution at pH 12.8, suggesting that depurination of TG residues by a glycosylase mechanism may occur. It is also postulated that TG residues are recognizable by the long-patch repair system, since UV-sensitive cells deficient for long-patch repair were more sensitive to TG than were wildtype cells. Furthermore, caffeine (1 mM) was shown to enhance the lethality of TG (3 microM), as monitored by colony formation, without altering levels of TG incorporation into DNA or the strand scission as measured immediately after treatment. This result, coupled with the known delayed cytotoxic response of TG, suggests that gaps may occur in newly synthesized DNA opposite TG residues and that the repair of these gaps by a postreplication repair mechanism is inhibited by caffeine.


Long-Term Observational Study of the Isolated Ostial Diagonal Stenosis in Patients with Chronic Coronary Syndrome.
Isolated ostial diagonal stenoses are very rare lesions in which percutaneous intervention could cause significant vessel compromise, and the long-term results have been reported in a few studies. This study sought the characteristics and long-term follow-up of the patients with isolated osteal diagonal stenosis regarding percutaneous coronary intervention and presence of angina. The study was an observational retrospective study conducted between January 2014 and December 2020. A total of 9769 patients who underwent coronary angiography were analyzed, and 87 patients had isolated diagonal stenosis. The patients were evaluated according to treatment modality and angina severity in long-term pattern. Median follow-up time was 36 months. A total of 54 (83.1%) patients were followed up with only medical treatment, and 11 (16.9%) patients underwent revascularization in addition to medical treatment. The degree of stenosis of the diagonal artery was significantly higher in the percutaneous coronary intervention group than medical group (P = 0.002) and the patients with wider reference diameter of diagonal artery complaint of more angina (P = 0.007). Class I angina was significantly higher in percutaneous coronary intervention group than medical and the patients with no angina were significantly higher in medical group than percutaneous coronary intervention group. Percutaneous coronary intervention was mainly performed for diagonal arteries with a higher degree of stenosis; however, the patients who underwent percutaneous coronary intervention had angina more than 50% rates. Furthermore, the patients with ongoing angina had a larger diameter of the diagonal artery regardless of the type of treatment.


Capecitabine in gastric cancer.
Chemotherapy is an important part of treatment for patients with gastric cancer. Although there is no single globally accepted standard of care for patients with advanced disease, regimens typically include a fluoropyrimidine and a platinum compound with or without a third drug (usually epirubicin or docetaxel). Oral fluoropyrimidines, such as capecitabine, offer clear advantages to patients in terms of convenience, but it is only recently that comprehensive data on their efficacy and safety in patients with gastric cancer have become available. The present article reviews capecitabine in the treatment of advanced and resectable gastric cancer. Ongoing Phase III trials involving capecitabine are also discussed. The data show that capecitabine is now established as an integral part of the multi-agent regimens used in the management of patients with gastric cancers.


Single Viruses on the Fluorescence Microscope: Imaging Molecular Mobility, Interactions and Structure Sheds New Light on Viral Replication.
Viruses are simple agents exhibiting complex reproductive mechanisms. Decades of research have provided crucial basic insights, antiviral medication and moderately successful gene therapy trials. The most infectious viral particle is, however, not always the most abundant one in a population, questioning the utility of classic ensemble-averaging virology. Indeed, viral replication is often not particularly efficient, prone to errors or containing parallel routes. Here, we review different single-molecule sensitive fluorescence methods that we employ routinely to investigate viruses. We provide a brief overview of the microscopy hardware needed and discuss the different methods and their application. In particular, we review how we applied (i) single-molecule Förster resonance energy transfer (smFRET) to probe the subviral human immunodeficiency virus (HIV-1) integrase (IN) quaternary structure; (ii) single particle tracking to study interactions of the simian virus 40 with membranes; (iii) 3D confocal microscopy and smFRET to quantify the HIV-1 pre-integration complex content and quaternary structure; (iv) image correlation spectroscopy to quantify the cytosolic HIV-1 Gag assembly, and finally; (v) super-resolution microscopy to characterize the interaction of HIV-1 with tetherin during assembly. We hope this review is an incentive for setting up and applying similar single-virus imaging studies in daily virology practice.


Differential Effects of Pesticide Applications on Liriomyza huidobrensis (Diptera: Agromyzidae) and its Parasitoids on Pea in Central Kenya.
Three Liriomyza species [Liriomyza huidobrensis (Blanchard), Liriomyza trifolii (Burgess), and Liriomyza sativae Blanchard] have been reported as the most important leafminer pests in vegetable production systems in Africa. In Kenya, farmers rely on indiscriminate synthetic insecticides use. On-farm field investigations were set up at three different locations (Sagana, Kabaru, and Naromoru) in central Kenya to determine the effect of pesticide application on the abundance of leafminers and their parasitoids under three management practices, namely: farmer practice (FP), reduced pesticide use (RP), and a control with no use of pesticides (CO). In addition, laboratory experiments were designed to test the effect of commonly used pesticides in pea production systems in central Kenya--Dimethoate, Dynamec, Thunder, Cyclone, Bestox, Folicur, Milraz, and Bulldock--on L. huidobrensis and two of its parasitoids, Diglyphus isaea Walker and Phaedrotoma scabriventris Nixon. The mean numbers of leafminer flies in control treatment were higher than in RP and FP in both first and second seasons across all sites, but RP and FP did not differ significantly. Parasitoid numbers were very low and there was no much variation between treatments at each location in both first and second seasons. No significant differences were observed between the three management practices with regards to the yield measurements. In the laboratory, the estimated LD50 values for L. huidobrensis larvae were all more than two times higher than the recommended dosages, while the LD50 of adults were below the recommended dosages. The estimated LD50 values for the parasitoids were much lower than recommended dosages for all pesticides except Thunder. This study, therefore, demonstrates that the pesticides currently used do not control the Liriomyza leafminer larvae that constitute the most destructive stage of the pest, but are rather detrimental to their parasitoids. In addition, the current low level of parasitoids recorded under field conditions even where no pesticide was used during this study, warrants consideration of classical biological control programs.


Hepatitis C virus infection suppresses the interferon response in the liver of the human hepatocyte chimeric mouse.
Recent studies indicate that hepatitis C virus (HCV) can modulate the expression of various genes including those involved in interferon signaling, and up-regulation of interferon-stimulated genes by HCV was reported to be strongly associated with treatment outcome. To expand our understanding of the molecular mechanism underlying treatment resistance, we analyzed the direct effects of interferon and/or HCV infection under immunodeficient conditions using cDNA microarray analysis of human hepatocyte chimeric mice. Human serum containing HCV genotype 1b was injected into human hepatocyte chimeric mice. IFN-α was administered 8 weeks after inoculation, and 6 hours later human hepatocytes in the mouse livers were collected for microarray analysis. HCV infection induced a more than 3-fold change in the expression of 181 genes, especially genes related to Organismal Injury and Abnormalities, such as fibrosis or injury of the liver (P = 5.90E-16∼3.66E-03). IFN administration induced more than 3-fold up-regulation in the expression of 152 genes. Marked induction was observed in the anti-fibrotic chemokines such as CXCL9, suggesting that IFN treatment might lead not only to HCV eradication but also prevention and repair of liver fibrosis. HCV infection appeared to suppress interferon signaling via significant reduction in interferon-induced gene expression in several genes of the IFN signaling pathway, including Mx1, STAT1, and several members of the CXCL and IFI families (P = 6.0E-12). Genes associated with Antimicrobial Response and Inflammatory Response were also significantly repressed (P = 5.22×10(-10)∼1.95×10(-2)). These results provide molecular insights into possible mechanisms used by HCV to evade innate immune responses, as well as novel therapeutic targets and a potential new indication for interferon therapy.


Penetration of moxifloxacin into rat mandibular bone and soft tissue.
Based on its in vitro activity and spectrum of activity, the new 8-methoxyquinolone antibiotic moxifloxacin (MXF) seems suited for the antibiotic therapy of odontogenic infections. Penetration into the relevant tissue is another prerequisite for clinical efficacy. For this reason, the levels of MXF in plasma, soft tissue, and mandibular bone were determined in an animal model with Wistar rats. Samples of 49 rats were analyzed. Tissue samples were homogenized and proteins were precipitated. The pharmacokinetic evaluation was conducted based on non-compartmental analysis. The concentration-time courses of tissues show a more plateau-shaped curve compared to plasma. Calculated AUC (area under the curve) ratios tissue:plasma were M. masseter:plasma = 2.64 and mandibles:plasma = 1.13. Administration of antibiotics is considered an important part of therapy during and/or after surgical procedures in the maxillofacial area. Because of the good penetration into bone and muscle tissues demonstrated in Wistar rats, MXF might be an option for clinical application in this indication.


PEGylated gene carriers in serum under shear flow.
The behaviour of drug/gene carriers in the blood stream under shear is still a puzzle. In this work, using the complexes formed by 21 bp DNA and poly(ethylene glycol)-b-poly(l-lysine) (PEG-PLL) of varying PEG lengths, we studied the dynamic behaviour of the complexes in the presence of fetal bovine serum (FBS) and under flow at different shear rates, a condition mimicking the internal physical environment of blood vessels. The PEG5k-PLL/DNA complex possesses a dense DNA/PLL core and a loose PEG5k protecting layer. The PEGylated DNA complexes exhibit multiple responses to external shear in the presence of FBS. The loose PEG5k layer is firstly disturbed at a shear rate below 30 s-1. The exposure of the charged core to the environment results in a secondary aggregation of the complex with FBS. The size of the aggregate is limited to a certain range as the shear rate increases to 50 s-1. The dense DNA/PLL core starts to withstand the shear force as the shear rate reaches 500 s-1. The reorganization of the core to accommodate more serum molecules leads to tertiary aggregation of the complexes. If PEG cannot form a valid layer around the complex, as in PEG2k-PLL/DNA, the complex forms an aggregate even without shear, and the first shear dependent region is missing. If the PEG layer is too stable around the complex, as in PEG10k-PLL/DNA, no tertiary aggregation occurs. The mechanism of shear on the behaviour of delivery particles in serum helps to design gene carriers with high efficacy.


Sources of Variation in Anesthetic Drug Costs.
Increasing attention has been focused on health care expenditures, which include anesthetic-related drug costs. Using data from 2 large academic medical centers, we sought to identify significant contributors to anesthetic drug cost variation. Using anesthesia information management systems, we calculated volatile and intravenous drug costs for 8 types of inpatient surgical procedures performed from July 1, 2009, to December 31, 2011. For each case, we determined patient age, American Society of Anesthesiologists (ASA) physical status, gender, institution, case duration, in-room provider, and attending anesthesiologist. These variables were then entered into 2 fixed-effects linear regression models, both with logarithmically transformed case cost as the outcome variable. The first model included duration, attending anesthesiologist, patient age, ASA physical status, and patient gender as independent variables. The second model included case type, institution, patient age, ASA physical status, and patient gender as independent variables. When all variables were entered into 1 model, redundancy analyses showed that case type was highly correlated (R = 0.92) with the other variables in the model. More specifically, a model that included case type was no better at predicting cost than a model without the variable, as long as that model contained the combination of attending anesthesiologist and case duration. Therefore, because we were interested in determining the effect both variables had on cost, 2 models were created instead of 1. The average change in cost resulting from each variable compared to the average cost of the reference category was calculated by first exponentiating the β coefficient and subtracting 1 to get the percent difference in cost. We then multiplied that value by the mean cost of the associated reference group. A total of 5504 records were identified, of which 4856 were analyzed. The median anesthetic drug cost was $38.45 (25th percentile = $23.23, 75th percentile = $63.82). The majority of the variation was not described by our models-35.2% was explained in the model containing case duration, and 32.3% was explained in the model containing case type. However, the largest sources of variation our models identified were attending anesthesiologist, case type, and procedure duration. With all else held constant, the average change in cost between attending anesthesiologists ranged from a cost decrease of $41.25 to a cost increase of $95.67 (10th percentile = -$19.96, 90th percentile = +$20.20) when compared to the provider with the median value for mean cost per case. The average change in cost between institutions was significant but minor ($5.73). The majority of the variation was not described by the models, possibly indicating high per-case random variation. The largest sources of variation identified by our models included attending anesthesiologist, procedure type, and case duration. The difference in cost between institutions was statistically significant but was minor. While many prior studies have found significant savings resulting from cost-reducing interventions, our findings suggest that because the overall cost of anesthetic drugs was small, the savings resulting from interventions focused on the clinical practice of attending anesthesiologists may be negligible, especially in institutions where access to more expensive drugs is already limited. Thus, cost-saving efforts may be better focused elsewhere.


Strongly Anisotropic Strain-Tunability of Excitons in Exfoliated ZrSe3.
The effect of uniaxial strain on the band structure of ZrSe3 , a semiconducting material with a marked in-plane structural anisotropy, is studied. By using a modified three-point bending test apparatus, thin ZrSe3 flakes are subjected to uniaxial strain along different crystalline orientations monitoring the effect of strain on their optical properties through micro-reflectance spectroscopy. The obtained spectra show excitonic features that blueshift upon uniaxial tension. This shift is strongly dependent on the direction along which the strain is being applied. When the flakes are strained along the b-axis, the exciton peak shifts at ≈60-95 meV %-1 , while along the a-axis, the shift only reaches ≈0-15 meV %-1 . Ab initio calculations are conducted to study the influence of uniaxial strain, applied along different crystal directions, on the band structure and reflectance spectra of ZrSe3 , exhibiting a remarkable agreement with the experimental results.


Aberrant popliteus muscle: anatomy and clinical consideration.
The aberrant popliteus muscle is an anomalous muscle in the popliteal fossa. It originates from the femur in the superior triangular surface of the popliteal fossa above the origin of the gastrocnemius muscle and it is inserted into the musculo- tendinous part on the medial side of the lateral head of the gastrocnemius muscle. The muscle lies immediately deep to the popliteal vein and during contraction is likely to compress the popliteal vein and reduce venous return from legs; it could also compress the lateral branches of the popliteal artery to the knee joint and the lateral head of the gastrocnemius muscle. It may present as a tumour of the popliteal fossa in radiographic examinations such as computed tomography (CT) and magnetic resonance imaging (MRI). If it undergoes hypertrophy as it may do in athletes and body builders, it may limit flexion of the knee joint, cause pain due to compression of surrounding structures and inhibit performance.


[Current data on the exploration and the evaluation of clinical pain].
Pain is a complex, multidimensional and multifactorial neuropsychological phenomenon with sensory-discriminative, affective, cognitive and behavioral components. Issues on pain assessment are related to the objective evaluation of a subjective phenomenon. Different levels of evaluation are to be considered. Some methods like visual analogue scale, numerical scale and verbal permit a global pain estimation. The aim of other methods like the McGill Pain Questionnaire is a quantitative and qualitative evaluation of the sensory and affective pain components. The measure of objective behavioral changes is an interesting approach, but at the present time there is no valid, simple and commonly used method. There is also a need for methods permitting a better exploration of a pain, and in particular a selective evaluation of organic and functional factors. The limits of psychological factors evaluation are reported.


Assessment of individual susceptibility to baseline DNA and cytogenetic damage in a healthy Turkish population: evaluation with lifestyle factors.
Cytogenetic biomarkers are most frequently used well-established endpoints in human population studies with their sensitivity for measuring exposure to genotoxic agents. They have an important role as early predictors of cancer risk. Identification of individual genotypes of metabolic gene polymorphisms helps to understand the modulation of cancer susceptibility by environmental exposures, such as cigarette smoking and other lifestyle factors. To evaluate individual susceptibility to chemicals, we determined individual DNA damage related to glutathione S-transferase (GST) genotypes (GSTM1, GSTT1, and GSTP1) in a Turkish population. Peripheral blood lymphocytes (PBL) and DNA samples of 127 subjects were analyzed for the presence of DNA damage, using single-cell gel electrophoresis (the Comet assay), and for cytogenetic parameters (chromosomal aberrations [CAs], bleomycin-induced CA, and a cytokinesis-blocked micronucleus assay), and the polymerase chain reaction/restriction fragment length polymorphism method, respectively. Individuals carrying a GSTT1-null allele showed higher frequencies of CA and micronucleus (MN) (p=0.026, p=0.003, respectively), whereas the GSTM1-null and GSTP1 mutant genotypes did not show any differences in cytogenetic parameters. Our findings demonstrated that none of the lifestyle factors (smoking, alcohol drinking, dietary habits, vitamin intake, and physical activity), except for vitamin intake (p=0.002), were significantly associated with the studied cytogenetic parameters. Our results suggest that the GSTT1 gene polymorphism may influence the baseline cytogenetic frequency in a healthy population.


Management of migraine headaches.
With 17% to 18% of women suffering from migraine headaches, clinicians will often be asked by their patients to prescribe medication. Migraine is an episodic chronic disease that is best managed with an overall treatment plan, rather than treated as an acute illness that is managed with sporadic medications. Realistic goals for the long-term management of migraine are based on patient education and an ongoing discussion between patient and provider. This article reviews the clinical presentation of migraine and recommendations for both acute and preventive treatment, including complementary therapies. "Red flags" that could be signs of more serious neurologic illness are presented. The management of migraine in pregnancy is also reviewed.


Potential role of gut microbes in the efficacy and toxicity of immune checkpoints inhibitors.
In recent years, Immune checkpoint inhibitors have been extensively used in the treatment of a variety of cancers. However, the response rates ranging from 13% to 69% depending on the tumor type and the emergence of immune-related adverse events have posed significant challenges for clinical treatment. As a key environmental factor, gut microbes have a variety of important physiological functions such as regulating intestinal nutrient metabolism, promoting intestinal mucosal renewal, and maintaining intestinal mucosal immune activity. A growing number of studies have revealed that gut microbes further influence the anticancer effects of tumor patients through modulation of the efficacy and toxicity of immune checkpoint inhibitors. Currently, faecal microbiota transplantation (FMT) have been developed relatively mature and suggested as an important regulator in order to enhance the efficacy of treatment. This review is dedicated to exploring the impact of differences in flora composition on the efficacy and toxicity of immune checkpoint inhibitors as well as to summarizing the current progress of FMT.


Association between the neuron-specific RNA-binding protein ELAVL4 and Parkinson disease.
Inflammatory processes have been implicated in the cascade of events that lead to nerve cell death. In the nervous system, a number of genes involved in inflammation pathways are regulated post-transcriptionally via the interaction of their mRNAs with specific RNA-binding Hu proteins, the vertebrate homologues of the Drosophila ELAV (for embryonic lethal abnormal vision). The gene encoding ELAVL4, a member of the Hu family of proteins, is located 2 Mb from the chromosome 1p linkage region peak for age-at-onset (AAO) of Parkinson disease (PD) (LOD = 3.41). Nine single-nucleotide polymorphisms (SNPs) in ELAVL4 were genotyped for 266 multiplex families (1,223 samples). Additional genotyping in 377 singleton families was performed for a subset of five SNPs (SNPs 1-5) that were not in linkage disequilibrium. SNP 2 (located in the first intron of ELAVL4) showed a strong significant association with AAO of PD (P = 0.006), and SNP 5 (a coding SNP in ELAVL4) showed a moderately significant association (P = 0.035). Haplotype analysis revealed that the A-C haplotype at SNPs 2 and 3 has the strongest significant association with AAO (P = 0.0001) among all combinations of two or three loci. The A-C haplotype remained significant for AAO after the inclusion of the C allele at SNP 5 to this haplotype (A-C-C haplotype, P = 0.00018). Although SNP 5 was found to associate with PD risk in the early-onset subset of PD families (at least one affected with AAO <40 years, 60 families), we believe that it is a by-product of its association with AAO. Taken together, these results suggest a potential role for ELAVL4 as a modifier gene for AAO of PD.


Promissory and protective imaginaries of regenerative medicine: Expectations work and scenario maintenance of disease research charities in the United Kingdom.
This article draws upon recent scholarship on technoscientific imaginaries and the sociology of technology expectations to reveal the mediating roles played by a number of disease-focused research charities in the United Kingdom. We examine the expectations they deal with about regenerative medicine research, and how they develop strategies to support and 'protect' potential medical scenarios for new therapies for dread diseases. In so doing, we develop and detail a concept of scenario maintenance to denote the strategic discursive and practical work of preserving stakeholders' faith in specific disease research pathways in the face of obstacles. Semi-structured in-depth interviews (N = 10) of research managers at nine research charities were qualitatively analysed, alongside a variety of charities' documentary data. Our analysis yielded three themes: managing and moderating media expectations; specifying expectations about disease-specific appropriateness of regenerative medicine; and maintaining scenarios of possible pathways for future success taking challenges into account.


[Effect of cerebral blood supply disorders on certain aspects of metabolism].
Bilateral ligation of common carotid arteries sharply decreases volume blood flow and oxygen consumption in the cortex, in diencephalon and midbrain whereas inducing no changes in the cerebellum and medulla oblongata. The same is true for intensity of metabolism of both total and separate fractions of phospholipids. This suggests that the changes of phospholipid metabolism in neural tissue depend on the degree of its blood supply disturbance and duration of the ischemia.


Tear film dynamics on an eye-shaped domain I: pressure boundary conditions.
We study the relaxation of a model for the human tear film after a blink on a stationary eye-shaped domain corresponding to a fully open eye using lubrication theory and explore the effects of viscosity, surface tension, gravity and boundary conditions that specify the pressure. The governing non-linear partial differential equation is solved on an overset grid by a method of lines using a finite-difference discretization in space and an adaptive second-order backward-difference formula solver in time. Our 2D simulations are calculated in the Overture computational framework. The computed flows show sensitivity to both our choices between two different pressure boundary conditions and the presence of gravity; this is particularly true around the boundary. The simulations recover features seen in 1D simulations and capture some experimental observations including hydraulic connectivity around the lid margins.


Altered gut microbiota promotes colitis-associated cancer in IL-1 receptor-associated kinase M-deficient mice.
Microbial sensing by Toll-like receptors (TLR) and its negative regulation have an important role in the pathogenesis of inflammation-related cancer. In this study, we investigated the role of negative regulation of Toll-like receptors signaling and gut microbiota in the development of colitis-associated cancer in mouse model. Colitis-associated cancer was induced by azoxymethane and dextran sodium sulfate in wild-type and in interleukin-1 receptor-associated kinase M (IRAK-M)-deficient mice with or without antibiotic (ATB) treatment. Local cytokine production was analyzed by multiplex cytokine assay or enzyme-linked immunosorbent assay, and regulatory T cells were analyzed by flow cytometry. Changes in microbiota composition during tumorigenesis were analyzed by pyrosequencing, and β-glucuronidase activity was measured in intestinal content by fluorescence assay. ATB treatment of wild-type mice reduced the incidence and severity of tumors. Compared with nontreated mice, ATB-treated mice had significantly lower numbers of regulatory T cells in colon, altered gut microbiota composition, and decreased β-glucuronidase activity. However, the β-glucuronidase activity was not as low as in germ-free mice. IRAK-M-deficient mice not only developed invasive tumors, but ATB-induced decrease in β-glucuronidase activity did not rescue them from severe carcinogenesis phenotype. Furthermore, IRAK-M-deficient mice had significantly increased levels of proinflammatory cytokines in the tumor tissue. We conclude that gut microbiota promotes tumorigenesis by increasing the exposure of gut epithelium to carcinogens and that IRAK-M-negative regulation is essential for colon cancer resistance even in conditions of altered microbiota. Therefore, gut microbiota and its metabolic activity could be potential targets for colitis-associated cancer therapy.


[Polymorphism of virus-like particles of retrotransposons in Drosophila and yeast cells].
Data on the molecular arrangement of viruslike particles (VLPs) of yeast and Drosophila retrotransposons are presented. Two methods for identifying VLPs from specific retrotransposon families have been offered. The first method is based on VLPs fractionation by electrophoresis in agarose gel under strictly controlled conditions. VLPs of the Drosophila melanogaster retrotransposon families copia and gypsy and D. virilis retrotransposon Tv1 were identified by this method. The method based on heterologous induction of retrotransposons in cells of the mutant spt3 strain of Saccharomyces cerevisiae was used to identify VLPs of yeast retrotransposon Tyl and D. melanogaster gypsy retrotransposon.


Mode coupling between first- and second-order whispering-gallery modes in coupled microdisks.
The mode characteristics for two coupled microdisks are investigated by the finite-difference time-domain technique. In the two coupled micodisks, mode coupling between the same order whispering-gallery modes (WGMs) results in coupled WGMs with split mode wavelengths. The numerical results show that the split mode wavelengths of the coupled first- and second-order WGMs can have a crossing point in some cases, which can induce anticrossing mode coupling between them and greatly reduce the mode Q factor of the coupled first-order WGMs. The time variation of mode field pattern shows the transformation between the coupled first- and second-order WGMs.


Sense and Sensibility: Characterizing Social Media Users Regarding the Use of Controversial Terms for COVID-19.
With the world-wide development of 2019 novel coronavirus, although WHO has officially announced the disease as COVID-19, one controversial term - "Chinese Virus" is still being used by a great number of people. In the meantime, global online media coverage about COVID-19-related racial attacks increases steadily, most of which are anti-Chinese or anti-Asian. As this pandemic becomes increasingly severe, more people start to talk about it on social media platforms such as Twitter. When they refer to COVID-19, there are mainly two ways: using controversial terms like "Chinese Virus" or "Wuhan Virus", or using non-controversial terms like "Coronavirus". In this article, we attempt to characterize the Twitter users who use controversial terms and those who use non-controversial terms. We use the Tweepy API to retrieve 17 million related tweets and the information of their authors. We find the significant differences between these two groups of Twitter users across their demographics, user-level features like the number of followers, political following status, as well as their geo-locations. Moreover, we apply classification models to predict Twitter users who are more likely to use controversial terms. To our best knowledge, this is the first large-scale social media-based study to characterize users with respect to their usage of controversial terms during a major crisis.


Constitutive G protein coupling profiles of understudied orphan GPCRs.
A large number of GPCRs are potentially valuable drug targets but remain understudied. Many of these lack well-validated activating ligands and are considered "orphan" receptors, and G protein coupling profiles have not been defined for many orphan GPCRs. Here we asked if constitutive receptor activity can be used to determine G protein coupling profiles of orphan GPCRs. We monitored nucleotide-sensitive interactions between 48 understudied orphan GPCRs and five G proteins (240 combinations) using bioluminescence resonance energy transfer (BRET). No receptor ligands were used, but GDP was used as a common G protein ligand to disrupt receptor-G protein complexes. Constitutive BRET between the same receptors and β-arrestins was also measured. We found sufficient GDP-sensitive BRET to generate G protein coupling profiles for 22 of the 48 receptors we studied. Altogether we identified 48 coupled receptor-G protein pairs, many of which have not been described previously. We conclude that receptor-G protein complexes that form spontaneously in the absence of guanine nucleotides can be used to profile G protein coupling of constitutively-active GPCRs. This approach may prove useful for studying G protein coupling of other GPCRs for which activating ligands are not available.


The New Role of Sirtuin1 in Human Osteoarthritis Chondrocytes by Regulating Autophagy.
The aim of this study is to investigate the role of Sirtuin1 (Sirt1) in the regulation of autophagy for human osteoarthritis (OA) chondrocytes. All cartilage samples were collected from human donors, including young group, aged group, and OA group. Primary chondrocytes were isolated and cultured with Sirt1 activator or inhibitor. Sirt1 expression in cartilage tissue and chondrocytes was evaluated, and the deacetylation activity of Sirt1 was determined. The alteration of autophagy activity after upregulating or downregulating Sirt1 was detected. Chondrocytes were treated with autophagy activator and inhibitor, and then the protein level of Sirt1 was examined. The interactions between Sirt1 and autophagy-related proteins Atg7, microtubule associated protein 1 light chain 3 (LC3), and Beclin-1 were determined by using immunoprecipitation. The assay of articular cartilage revealed that the expression of Sirt1 might be age-related: highly expressed in of younger people, and respectively decreased in the elderly people and OA patients. In vitro study was also validated this result. Further study confirmed that higher levels of Sirt1 significantly increased autophagy in aged chondrocytes, while the lower expression of Sirt1 reduced autophagy in young chondrocytes. Of note, the high levels of Sirt1 reduced autophagy in OA chondrocytes. When the chondrocytes were treated with autophagy activator or inhibitor, we found the expression of Sirt1 was not affected. In addition, we found that Sirt1 could interact with Atg7. These results suggest that Sirt1 in human chondrocytes regulates autophagy by interacting with autophagy related Atg7, and Sirt1 may become a more important target in OA treatment.


Casemix funding for acute hospital inpatient services in Australia.
Casemix funding was introduced first in Victoria in 1993-94, and since then most States have moved towards either casemix funding or using casemix to inform the budget setting process. The five States implementing casemix have adopted some common funding elements: all use AN-DRG-3; all have introduced capping, msot commonly at the hospital level; and all ensure accuracy of diagnosis and procedure coding through coding audits. Two funding models have been developed. The fixed and variable model involves a fixed grant for hospital overhead costs and a payment for each patient treated, covering only variable costs. The integrated model provides an integrated payment to hospitals for each patient treated, covering both the fixed and variable costs. There are different weight setting processes and base prices between the States, which result in marked differences in the price paid for the same type of case treated in similar hospitals. Learning across State boundaries should be encouraged, with knowledge of what is effective and what is ineffective in casemix funding arrangements being used to develop Australian best practice in this area.


A novel Ka-band chirped-pulse spectrometer used in the determination of pressure broadening coefficients of astrochemical molecules.
A novel chirped-pulse Fourier transform microwave (CP-FTMW) spectrometer has been constructed to cover the Ka-band (26.5 GHz-40 GHz) for use in the CRESUCHIRP project, which aims to study the branching ratios of reactions at low temperatures using the chirped-pulse in uniform flow technique. The design takes advantage of recent developments in radio-frequency components, notably, high-frequency, high-power solid-state amplifiers. The spectrometer had a flatness of 5.5 dB across the spectral range, produced harmonic signals below -20 dBc, and the recorded signal scaled well to 6 × 106 averages. The new spectrometer was used to determine pressure broadening coefficients with a helium collider at room temperature for three molecules relevant to astrochemistry, applying the Voigt function to fit the magnitude of the Fourier-transformed data in the frequency domain. The pressure broadening coefficient for carbonyl sulfide was determined to be (2.45 ± 0.02) MHz mbar-1 at room temperature, which agreed well with previous measurements. Pressure broadening coefficients were also determined for multiple transitions of vinyl cyanide and benzonitrile. Additionally, the spectrometer was coupled with a cold, uniform flow from a Laval nozzle. The spectrum of vinyl cyanide was recorded in the flow, and its rotational temperature was determined to be (24 ± 11) K. This temperature agreed with a prediction of the composite temperature of the system through simulations of the experimental environment coupled with calculations of the solution to the optical Bloch equations. These results pave the way for future quantitative studies in low-temperature and high-pressure environments using CP-FTMW spectroscopy.


Increased Osmolarity in Biofilm Triggers RcsB-Dependent Lipid A Palmitoylation in Escherichia coli.
Biofilms are often described as protective shelters that preserve bacteria from hostile surroundings. However, biofilm bacteria are also exposed to various stresses and need to adjust to the heterogeneous physicochemical conditions prevailing within biofilms. In Gram-negative bacteria, such adaptations can result in modifications of the lipopolysaccharide, a major component of the outer membrane characterized by a highly dynamic structure responding to environmental changes. We previously showed that Gram-negative biofilm bacteria undergo an increase in lipid A palmitoylation mediated by the PagP enzyme, contributing to increased resistance to host defenses. Here we describe a regulatory pathway leading to transcriptional induction of pagP in response to specific conditions created in the biofilm environment. We show that pagP expression is induced via the Rcs envelope stress system independently of the Rcs phosphorelay cascade and that it requires the GadE auxiliary regulator. Moreover, we identify an increase in osmolarity (i.e., ionic stress) as a signal able to induce pagP expression in an RcsB-dependent manner. Consistently, we show that the biofilm is a hyperosmolar environment and that RcsB-dependent pagP induction can be dampened in the presence of an osmoprotectant. These results provide new insights into the adaptive mechanisms of bacterial differentiation in biofilm.IMPORTANCE The development of the dense bacterial communities called biofilms creates a highly heterogeneous environment in which bacteria are subjected to a variety of physicochemical stresses. We investigated the mechanisms of a widespread and biofilm-associated chemical modification of the lipopolysaccharide (LPS), a major component of all Gram-negative bacterial outer membranes. This modification corresponds to the incorporation, mediated by the enzyme PagP, of a palmitate chain into lipid A (palmitoylation) that reduces bacterial recognition by host immune responses. Using biochemical and genetic approaches, we demonstrate that a significant part of biofilm-associated lipid A palmitoylation is triggered upon induction of pagP transcription by the hyperosmolar biofilm environment. pagP induction is regulated by RcsB, the response regulator of the Rcs stress response pathway, and is not observed under planktonic conditions. Our report provides new insights into how physiological adaptations to local biofilm microenvironments can contribute to decreases in susceptibility to antimicrobial agents and host immune defenses.


[Estimating utility values for health status using the Spanish version of the SF-36. Validity of the SF-6D index vs EQ-5D].
A new utility index derived of the SF-36, the SF-6D, was recently developed and has been compared with other utility measures. The objective was to validate this index in the Spanish version of SF-36 with respect to the EQ-5D. 1,843 complete measures of the SF-36 and the EQ-5D from 1,283 patients who received a solid organ transplant were collected. SF-6D values were calculated using the model proposed by its creator and without tariff values (not weighted). EQ-5D values were calculated using Spanish visual analog scale tariff (VAS-t), the time-trade off tariff (TTO-t) and also without tariff values (not weighted). Spearman correlation coefficients were calculated between SF-6D and EQ-5D values. Mean value (standard deviation) of SF-6D was 0.67 (0.15), of VAS-t, 0.69 (0.24) and of TTO-t, 0.70 (0.32). SF-6D values had moderate correlation with VAS-t (r = 0.734) and TTO-t (r = 0.731) (both p < 0.001). The not weighted SF-6D had a high correlation with the weighted version (r = 0.969, p < 0.001), and moderate with the VAS-t (r = 0.754), TTO-t (r = 0.750) and no weighted EQ-5D (r = 0.784) (p < 0.001). The SF-6D index derived from the Spanish version of SF-36 seem to be a valid utility index to be used with the SF-36 databases from studies made in Spain.


The Impact of Adjuvant Postoperative Radiation Therapy and Chemotherapy on Survival After Esophagectomy for Esophageal Carcinoma.
The objective of this study was to analyze the impact on overall survival (OS) from the addition of postoperative radiation with or without chemotherapy after esophagectomy, using a large, hospital-based dataset. Previous retrospective studies have suggested an OS advantage for postoperative chemoradiation over surgery alone, although prospective data are lacking. The National Cancer Data Base was queried to select patients diagnosed with stage pT3-4Nx-0M0 or pT1-4N1-3M0 esophageal carcinoma (squamous cell or adenocarcinoma) from 1998 to 2011 treated with definitive esophagectomy ± postoperative radiation and/or chemotherapy. OS was analyzed using the Kaplan-Meier method and compared using the log-rank test. Multivariate Cox regression analysis was used to identify covariates associated with OS. There were 4893 patients selected, of whom 1153 (23.6%) received postoperative radiation. Most patients receiving radiation also received sequential/concomitant chemotherapy (89.9%). For the entire cohort, postoperative radiation was associated with a statistically significant but modest absolute improvement in survival (hazard ratio 0.77; 95% CI, 0.71-0.83; P < 0.001). On subgroup analysis, postoperative radiation was associated with improved OS for patients with node-positive disease (3-yr OS 34.3 % vs 27.8%, P < 0.001) or positive margins (3-yr OS 36.4% vs 18.0%, P < 0.001). When chemotherapy usage was incorporated, sequential chemotherapy was associated with the best survival (P < 0.001). Multivariate analysis revealed that the addition of chemotherapy to radiation therapy, whether sequentially or concurrently, was a strong prognostic factor for OS. In this hospital-based study, the addition of postoperative chemoradiation (either sequentially or concomitantly) after esophagectomy was associated with improved OS for patients with node-positive disease or positive margins.


Association of Coronary Artery Severity and Late In-Stent Restenosis: An Angiographic Imaging Study.
Coronary in-stent restenosis (ISR) remains a challenge in interventional cardiology. We investigated the relationship between angiographic pre-interventional grade of lesion stenosis (LS) and the prognosis of late ISR. After exclusions, 110 patients with ISR and 109 patients without ISR were compared. In the ISR group, the grade of LS was greater (P < .001) and the length of the critical segment (LCS) was longer (P < .001). Stent length was longer in the ISR group (P = .008). Compared with the LCS, the grade of LS above 87.5% is 6.9 times more predictive of ISR than the LCS >10.5 mm. Kaplan-Meier curve analysis showed that the grade of initial LS >87.5% had a higher ISR rate than the grade of LS <87.5% (log-rank test P < .001) and critical lesion length over 10.5 mm had a higher ISR rate than critical lesion length under 10.5 mm (log-rank test P < .001). The present study found that the angiographic pre-interventional grades of LS and LCS were important predictors of ISR. Pre-interventional angiographic stenosis >87.5% was significantly predictive of late ISR.


μ-Opioid receptor availability in the amygdala is associated with smoking for negative affect relief.
The perception that smoking relieves negative affect contributes to smoking persistence. Endogenous opioid neurotransmission, and the μ-opioid receptor (MOR) in particular, plays a role in affective regulation and is modulated by nicotine. We examined the relationship of MOR binding availability in the amygdala to the motivation to smoke for negative affect relief and to the acute effects of smoking on affective responses. Twenty-two smokers were scanned on two separate occasions after overnight abstinence using [¹¹C]carfentanil positron emission tomography imaging: after smoking a nicotine-containing cigarette and after smoking a denicotinized cigarette. Self-reports of smoking motives were collected at baseline, and measures of positive and negative affect were collected pre- and post- cigarette smoking. Higher MOR availability in the amygdala was associated with motivation to smoke to relieve negative affect. However, MOR availability was unrelated to changes in affect after smoking either cigarette. Increased MOR availability in amygdala may underlie the motivation to smoke for negative affective relief. These results are consistent with previous data highlighting the role of MOR neurotransmission in smoking behavior.


Stimulus-dependent modulation of human B cell function by T cell clones.
T cells exert both positive and negative regulatory effects on B cell function. To determine whether the nature of the stimulus could alter the immunoregulatory effects of T cells, the capacity of a battery of human T cell clones to modulate B cell function after stimulation with either pokeweed mitogen or a mAb to the CD3 molecular complex was examined. It was observed that most clones induced B cell differentiation when stimulated with immobilized mAb to CD3 (64.1) regardless of their phenotype. Moreover, the majority of clones (42 of 51) augmented the generation of immunoglobulin-secreting cells (ISC) supported by anti-CD3-stimulated blood CD4+ T cells. By contrast, none of the clones induced B cell differentiation when stimulated with PWM and 48 of 51 clones suppressed the generation of ISC induced by blood CD4+ T cells. This suppression could not be accounted for by the depletion of essential molecules or factors or by secretion of suppressive factors. Suppressive activity of clones did not correlate with the CD4 or CD8 phenotype and was not overcome by the addition of supernatants generated from mitogen-stimulated T cells or recombinant IL-2. Suppression by most clones, however, was abrogated when the clones were treated with mitomycin C or irradiated. A number of suppressive mechanisms by individual PWM-stimulated clones was identified, including direct inhibition of B cell function by cytotoxic and non-lytic means and suppression of helper T cell function. The failure of anti-CD3-stimulated clones to suppress the differentiation of B cells appeared to reflect the capacity of this stimulus to induce apoptosis by the clones after initial activation. These results indicate that T cell clones can provide help for B cell differentiation or can suppress B cell function by a variety of mechanisms depending upon the mode of stimulation.


An Examination of Chronic Pain Indices and the Updated Posttraumatic Stress Disorder Checklist for Diagnostic and Statistical Manual of Mental-Disorders-Fifth Edition.
Chronic pain and post-traumatic stress disorder (PTSD) comorbidity is prevalent among veterans and is associated with increased levels of pain severity and pain-related disability. An improved understanding of the relationship between these co-occurring disorders, in addition to effective integrated treatments, will develop by considering the changes to the PTSD diagnostic criteria in the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5). The current study examined the relationship between the revised PTSD Checklist for DSM-5 (PCL-5) symptom clusters (i.e., intrusion, avoidance, negative alterations in cognition and mood [NACM], and arousal) and chronic pain measurements (i.e., pain severity, interference, and disability). Participants included 103 veterans (ages 26-70, mean = 45.33) participating in a randomized clinical trial examining the efficacy of an interdisciplinary pain management program for chronic musculoskeletal pain. The study was approved by a university system Institutional Review Board and affiliated healthcare system. The participants with a provisional PTSD diagnosis based on PCL-5 responses (N = 76) had significantly greater pain severity, interference, and disability than the participants without a provisional diagnosis (N = 23). Correlations between symptom clusters and pain measurements were mostly significant and positive with varying strengths. The avoidance symptom cluster, however, had relatively weaker correlations with pain measurements and was not significantly associated with the numeric rating scale of pain severity. Path analyses revealed that, after controlling for avoidance symptoms, significant associations remained between NACM and all the pain measurements. After controlling for NACM symptoms, however, there were no significant associations between avoidance symptoms and pain measurements. The current study highlights a need to re-examine the leading theories about the mutual maintenance of these disorders in order to develop effective integrative treatment approaches. PTSD-related avoidance may have a relatively weaker role in co-occurring chronic pain than the other symptom clusters and may have a qualitatively different role than chronic pain-related avoidance. Future research should explore the relationship between the avoidance in PTSD and the avoidance in chronic pain as well as identify which chronic pain measurements are the most useful when examining the relationship between PTSD and chronic pain. The potential impact of trauma-related cognition and mood on chronic pain indicates that this is an important area for intervention and should be considered in the development of integrated treatments for chronic pain and PTSD among veterans.


Identification of V3 loop-binding proteins as potential receptors implicated in the binding of HIV particles to CD4(+) cells.
The binding of human immunodeficiency virus (HIV) type 1 particles to CD4(+) cells could be blocked either by antibodies against the V3 loop domain of the viral external envelope glycoprotein gp120, or by the V3 loop mimicking pseudopeptide 5[Kpsi(CH2N)PR]-TASP, which forms a stable complex with a cell-surface-expressed 95-kDa protein. Here, by using an affinity matrix containing 5[Kpsi(CH2N)PR]-TASP and cytoplasmic extracts from human CEM cells, we purified three V3 loop-binding proteins of 95, 40, and 30 kDa, which after microsequencing were revealed to be as nucleolin, putative HLA class II-associated protein (PHAP) II, and PHAP I, respectively. The 95-kDa cell-surface protein was also isolated and found to be nucleolin. We show that recombinant preparations of gp120 bind the purified preparations containing the V3 loop-binding proteins with a high affinity, comparable to the binding of gp120 to soluble CD4. Such binding is inhibited either by 5[Kpsi(CH2N)PR]-TASP or antibodies against the V3 loop. Moreover, these purified preparations inhibit HIV entry into CD4(+) cells as efficiently as soluble CD4. Taken together, our results suggest that nucleolin, PHAP II, and PHAP I appear to be functional as potential receptors in the HIV binding process by virtue of their capacity to interact with the V3 loop of gp120.


Discovery of a novel small molecule inhibitor targeting the frataxin/ubiquitin interaction via structure-based virtual screening and bioassays.
Friedreich's ataxia (FRDA) is an autosomal recessive neuro- and cardiodegenerative disorder for which there are no proven effective treatments. FRDA is caused by decreased expression and/or function of the mitochondrial protein frataxin. Here, we report findings that frataxin is degraded via the ubiquitin-proteasomal pathway and that it is ubiquitinated at residue K(147) in Calu-6 cells. A theoretical model of the frataxin-K(147)/Ub complex, constructed by combining bioinformatics interface predictions with information-driven docking, revealed a hitherto unnoticed, potential ubiquitin-binding domain in frataxin. Through structure-based virtual screening and cell-based assays, we discovered a novel small molecule (compound (+)-11) able to prevent frataxin ubiquitination and degradation. (+)-11 was synthesized and tested for specific binding to frataxin by an UF-LC/MS based ligand-binding assay. Follow-up scaffold-based searches resulted in the identification of a lead series with micromolar activity in disrupting the frataxin/Ub interaction. This study also suggests that frataxin could be a potential target for FRDA drug development.


A spectrum of molecular variation in a cohort of Italian families with trimethylaminuria: identification of three novel mutations of the FM03 gene.
Fish-odor syndrome or trimethylaminuria, is a rare inborn error of metabolism inherited in an autosomal recessive fashion, involving the dysfunction of hepatic enzyme flavin-containing monooxygenase 3 (FMO3) that converts fishy-smelling trimethylamine (TMA) into odorless trimethylamine-N-oxide (TMAO). This confers, to the affected individual a very unpleasant body odor resembling that of rotting fish. This disorder has been relatively well-documented in British, Australian, and American populations and reports have appeared regarding patients in Thailand and Hong Kong, but no Italian families affected by trimethylaminuria have been reported in the literature. We have collected a cohort of Italian families and investigated the genetic basis of the disorder in these Italian pedigrees disclosing a spectrum of molecular variation in the FM03 gene comprising three novel deleterious mutations: the first documented de novo missense mutation causative of trimethylaminuria; a guanidine nucleotide deletion (G1182del) at codon 394 and a novel missense mutation (R238P) that altered highly conserved amino acid in the exon 6. Moreover, we investigated by aplotype analysis a family with mild TMAuria identifying a putative causative aplotype. Finally, we failed to detect any variation in other Italian families suggesting that this gene is not associated with all clinical form of trimethylaminuria or that polymorphisms in this gene could be susceptibility factors for developing the disease. Our findings support the hypothesis that TMAuria is not a rare recessive disorder but rather a spectrum of malodour phenotypes in which diet and environmental exposures can play a role in triggering symptoms.


AZGP1 Protein Expression in Hormone-Naïve Advanced Prostate Cancer Treated with Primary Androgen Deprivation Therapy.
Biomarkers for predicting the risk of castration-resistant prostate cancer (CRPC) in men treated with primary androgen deprivation therapy (ADT) are lacking. We investigated whether Zinc-alpha 2 glycoprotein (AZGP1) expression in the diagnostic biopsies of men with hormone-naïve prostate cancer (PCa) undergoing primary ADT was predictive of the development of CRPC and PCa-specific mortality. The study included 191 patients who commenced ADT from 2000 to 2011. The AZGP1 expression was evaluated using immunohistochemistry and scored as high or low expression. The risks of CRPC and PCa-specific mortality were analyzed using stratified cumulative incidences and a cause-specific COX regression analysis for competing risk assessment. The median follow-up time was 9.8 (IQR: 6.1-12.7) years. In total, 94 and 97 patients presented with low and high AZGP1 expression, respectively. A low AZGP1 expression was found to be associated with a shorter time to CRPC when compared to patients with a high AZGP1 expression (HR: 1.5; 95% CI: 1.0-2.1; p = 0.03). However, the multivariable analysis demonstrated no added benefit by adding the AZGP1 expression to prediction models for CRPC. No differences for PCa-specific mortality between the AZGP1 groups were observed. In conclusion, a low AZGP1 expression was associated with a shorter time to CRPC for PCa patients treated with first-line ADT but did not add any predictive information besides well-established clinicopathological variables.


Platforms to test the temporospatial capabilities of carrier systems in delivering growth factors to benefit vascular bioengineering.
In this study we produced a set of in vitro culture platforms to model vascular cell responses to growth factors and factor delivery vehicles. Two of the systems (whole vessel and whole lung vascular development) were supported by microfluidic systems facilitating media circulation and waste removal. We assessed vascular endothelial growth factor (VEGF) delivery by Pluronic F-127 hydrogel, 30 nm pore-sized microparticles (MPs), 60 nm pore-sized MP or a 50/50 mixture of 30 and 60 nm pore-sized MP. VEGF was delivered to porcine acellular lung vascular scaffolds (2.5 cm2 square pieces or whole 3D segments of acellular blood vessels) as well as whole acellular lung scaffolds. Scaffold-cell attachment was examined as was vascular tissue formation. We showed that a 50/50 mixture of 30 and 60 nm pore-sized silicon wafer MPs allowed for long-term release of VEGF within the scaffold vasculature and supported vascular endothelial tissue development during in vitro culture.


Development of Multilocus PCR Assays for Raffaelea lauricola, Causal Agent of Laurel Wilt Disease.
Laurel wilt, caused by the fungus Raffaelea lauricola, is an exotic disease that affects members of the Lauraceae plant family in the southeastern United States. The disease is spreading rapidly in native forests and is now found in commercial avocado groves in south Florida, where an accurate diagnostic method would improve disease management. A polymerase chain reaction (PCR) method based on amplifying the ribosomal small-subunit DNA, with a detection limit of 0.0001 ng, was found to be suitable for some quantitative PCR applications; however, it was not taxon specific. Genomic sequencing of R. lauricola was used to identify and develop primers to amplify two taxon-specific simple-sequence repeat (SSR) loci, which did not amplify from related taxa or host DNA. The new SSR loci PCR assay has a detection limit of 0.1 ng of R. lauricola DNA, is compatible with traditional and real-time PCR, was tested in four labs to confirm consistency, and reduces diagnostic time from 1 week to 1 day. Our work illustrates pitfalls to designing taxon-specific assays for new pathogens and that undescribed fungi can limit specificity.


Association of Sensory Liking for Fat with Dietary Intake and Metabolic Syndrome in Korean Adults.
Individual sensory liking is perceived as a major determinant of dietary intake and may influence chronic disease. This study aimed to assess the odds of metabolic syndrome in Korean adults based on their liking for fat. Data from 7731 adults aged 40⁻69, included in the Korean Genome and Epidemiology Study, were collected. Fat liking scores were obtained from self-report questionnaires. In both genders, sensory liking for fat was positively associated with consumption of red meat and added fat. Subjects with a stronger liking for fat showed a higher intake of energy and fat and a lower intake of vitamin C and fiber as compared to subjects with a lower liking for fat. There were increasing trends in the odds of metabolic syndrome with stronger liking for fat (odds ratios (ORs) for the Like group compared to the Dislike group, men: ORs = 1.29 (95% confidence interval 1.06⁻1.50) p-trend = 0.01; women: ORs = 1.28 (1.04⁻1.58) p-trend = 0.018) after adjustment for age, alcohol intake, smoking, exercise, education level, and income status. Our results suggested that the liking for fat-rich food might partially contribute to the increased odds of metabolic syndrome.


Cooperative loading of multisite receptors with lanthanide containers: an approach for organized luminescent metallopolymers.
Metal-containing (bio)organic polymers are materials of continuously increasing importance for applications in energy storage and conversion, drug delivery, shape-memory items, supported catalysts, organic conductors and smart photonic devices. The embodiment of luminescent components provides a revolution in lighting and signaling with the ever-increasing development of polymeric light-emitting devices. Despite the unique properties expected from the introduction of optically and magnetically active lanthanides into organic polymers, the deficient control of the metal loading currently limits their design to empirical and poorly reproducible materials. We show here that the synthetic efforts required for producing soluble multi-site host systems Lk are largely overcome by the virtue of reversible thermodynamics for mastering the metal loading with the help of only two parameters: (1) the affinity of the luminescent lanthanide container for a single binding site and (2) the cooperative effect which modulates the successive fixation of metallic units to adjacent sites. When unsymmetrical perfluorobenzene-trifluoroacetylacetonate co-ligands (pbta-) are selected for balancing the charge of the trivalent lanthanide cations, Ln3+, in six-coordinate [Ln(pbta)3] containers, the explored anti-cooperative complexation processes induce nearest-neighbor intermetallic interactions twice as large as thermal energy at room temperature (RT = 2.5 kJ mol-1). These values have no precedent when using standard symmetrical containers and they pave the way for programming metal alternation in luminescent lanthanidopolymers.


Cross-sectional areas of rotator cuff muscles in males without tears on shoulder MRI.
To establish reference values of rotator cuff (RC) cross sectional area (CSA) in males. We retrospectively analyzed shoulder MRIs from 500 patients aged 13-78 years, grouped as follows (N=100 in each): <20, 20-30, 30-40, 40-50, >50 years. All examinations were reviewed to exclude prior surgery, tears, or significant RC pathology. We segmented a standardized T1 sagittal MR image in each case to obtain CSA of supraspinatus (SUP), infraspinatus/teres minor (INF), and subscapularis (SUB) muscles. Across age groups, we recorded individual and total muscle CSA. We also performed ratios between individual muscle CSA and total CSA to examine total muscle mass contribution over age groups. We tested for differences between age groups controlled for BMI. CSAs for SUP, INF, SUB, and total RC CSA were lower in subjects >50 years compared to all other groups (P<0.003 for all comparisons), persisting after controlling for BMI (P<0.03). Relative contribution of SUP CSA to total RC CSA was stable across age groups (P>0.32). INF CSA relative to total RC CSA increased with age, whereas SUB decreased (P<0.005). Subjects >50 years showed lower SUP (-15%), INF (-6%), and SUB (-21%) CSA, when compared to mean CSAs of all subjects <50 years. Total RC CSA significantly correlated with age (r=-0.34, P<0.001), persisting after controlling for BMI (r=-0.42, P<0.001). RC muscles in male subjects with no tears on MRI show decreasing CSA with age, independent of BMI.


Differential Impact of the Pinewood Nematode on Pinus Species Under Drought Conditions.
The pinewood nematode (PWN), Bursaphelenchus xylophilus, responsible for the pine wilt disease (PWD), is a major threat to pine forests worldwide. Since forest mortality due to PWN might be exacerbated by climate, the concerns regarding PWD in the Mediterranean region are further emphasized by the projected scenarios of more drought events and higher temperatures. In this context, it is essential to better understand the pine species vulnerability to PWN under these conditions. To achieve that, physiological responses and wilting symptoms were monitored in artificially inoculated Pinus pinaster (P. pinaster), Pinus pinea (P. pinea), and Pinus radiata (P. radiata) saplings under controlled temperature (25/30°C) and water availability (watered/water stressed). The results obtained showed that the impact of PWN is species-dependent, being infected P. pinaster and P. radiata more prone to physiological and morphological damage than P. pinea. For the more susceptible species (P. pinaster and P. radiata), the presence of the nematode was the main driver of photosynthetic responses, regardless of their temperature or water regime conditions. Nevertheless, water potential was revealed to be highly affected by the synergy of PWN and the studied abiotic conditions, with higher temperatures (P. pinaster) or water limitation (P. radiata) increasing the impact of nematodes on trees' water status. Furthermore, water limitation had an influence on nematodes density and its allocation on trees' structures, with P. pinaster revealing the highest nematode abundance and inner dispersion. In inoculated P. pinea individuals, nematodes' population decreased significantly, emphasizing this species resistance to PWN. Our findings revealed a synergistic impact of PWN infection and stressful environmental conditions, particularly on the water status of P. pinaster and P. radiata, triggering disease symptoms and mortality of these species. Our results suggest that predicted drought conditions might facilitate proliferation and exacerbate the impact of PWN on these two species, through xylem cavitation, leading to strong changes in pine forests of the Mediterranean regions.


Species Identification and In Vitro Antifungal Susceptibility of Paecilomyces/Purpureocillium Species Isolated from Clinical Respiratory Samples: A Multicenter Study.
Paecilomyces spp. are emerging fungal pathogens, where Paecilomyceslilacinus and Paecilomyces variotii are the most reported species. Taxonomic and phylogenetic revisions in this genus have shown that P. variotii represents a species complex, whereas P. lilacinus is related to another genus called Purpureocillium. The aims of this study were to identify clinical isolates of Paecilomyces spp. at the species level, and to determine their antifungal susceptibility profiles. 70 clinical Paecilomyces spp. isolates were identified by MALDI-TOF Mass Spectrometry (MS) and by multilocus rDNA genes sequencing including ITS and the D1/D2 genes. Among the 70 Paecilomyces spp. isolates, 28 were identified as P. lilacinum, 26 as P. variotii stricto sensu, and 16 as P. maximus. For antifungal susceptibility testing, Minimal Inhibitory Concentrations (MICs) or Minimal Effective Concentrations (MECs) were determined for 8 antifungals. All P. lilacinum isolates had high MICs and MECs of amphotericin B and echinocandins, respectively, unlike P. variotii and P. maximus. For azole drugs, MICs were molecule- and species- dependent. The differences in in vitro susceptibility to antifungals underline the importance of accurate species identification. The MALDI-TOF MS can be a good alternative in routine laboratory to ensure fast identification of Paecilomyces spp. and P. lilacinum.


Causes and Consequences of Metabolic Acidosis in Patients after Kidney Transplantation.
Metabolic acidosis (MA) is a common complication in kidney transplantation (KTx). It is more prevalent in KTx than in CKD, and it occurs at higher glomerular filtration rates. The pathophysiologic understanding of MA in KTx and its clinical impact has been highlighted by few recent studies. However, no guidelines exist yet for the treatment of MA after KTx. MA in KTx seems to share pathophysiologic mechanisms with CKD, such as impaired ammoniagenesis. Additional kidney transplant-specific factors seem to alter not only the prevalence but also the phenotype of MA, which typically shows features of renal tubular acidosis. There is evidence that calcineurin inhibitors, immunological factors, process of donation, donor characteristics, and diet may contribute to MA occurrence. According to several mainly observational studies, MA seems to play a role in disturbed bone metabolism, cardiovascular morbidity, declining graft function, and mortality. A better understanding of the pathophysiology and evidence from randomized controlled trials, in particular, are needed to clarify the role of MA and the potential benefit of alkali treatment in KTx. Alkali therapy might not only be beneficial but also cost effective and safe. Key Messages: MA seems to be associated with several negative outcomes in KTx. A deeper understanding of the pathophysiology and clinical consequences of MA in KTx is crucial. Clinical trials will have to determine the potential benefits of alkali therapy.


Pathology consultation. Extracranial meningiomas.
Extracranial meningiomas are most often due to extension from an intracranial primary and less so from meningocytic cells found with arachnoid clusters or modified mesenchymal cells. The most common presentation of all forms is cutaneous, either primary or secondary. Orbital, paranasal, temporal bone and oral presentations follow in that order.


Norstictic Acid Is a Selective Allosteric Transcriptional Regulator.
Inhibitors of transcriptional protein-protein interactions (PPIs) have high value both as tools and for therapeutic applications. The PPI network mediated by the transcriptional coactivator Med25, for example, regulates stress-response and motility pathways, and dysregulation of the PPI networks contributes to oncogenesis and metastasis. The canonical transcription factor binding sites within Med25 are large (∼900 Å2) and have little topology, and thus, they do not present an array of attractive small-molecule binding sites for inhibitor discovery. Here we demonstrate that the depsidone natural product norstictic acid functions through an alternative binding site to block Med25-transcriptional activator PPIs in vitro and in cell culture. Norstictic acid targets a binding site comprising a highly dynamic loop flanking one canonical binding surface, and in doing so, it both orthosterically and allosterically alters Med25-driven transcription in a patient-derived model of triple-negative breast cancer. These results highlight the potential of Med25 as a therapeutic target as well as the inhibitor discovery opportunities presented by structurally dynamic loops within otherwise challenging proteins.


Etiology and pathogenesis of type 1 diabetes.
An increasing bulk of evidence suggests that type 1 (insulin-dependent) diabetes mellitus is an autoimmune disease with a strong immunogenetic background. 1st-degree relatives of type 1 diabetic patients, especially HLA-identical individuals, bear an increased risk to develop the disease. The autoimmune reactions are pronounced at the onset of disease where an infiltration of islets with T and B lymphocytes, plasma cells and macrophages can be observed. Autoreactive T lymphocytes play a crucial role among effector mechanisms which finally lead to a selective destruction of pancreatic beta cells. Disease-specific autoantibodies (Ab) include cytoplasmic islet cell Ab (ICA), islet cell surface Ab (ICSA), Ab to the 64KD islet cell protein and Ab to insulin (IAA). As ICA can be detected months or years before the onset of clinical disease, testing of individuals at risk or population screening programs can help to recognize subclinical insulitis. High titers of ICA and high levels of IAA, as measured by radioimmunoassay, indicate a high risk for progression to type 1 diabetes. A blunted first phase insulin response in the i.v. glucose tolerance test is the most sensitive sign of an irreversible metabolic deterioration. It is likely that immunotherapy at a prediabetic state will be more efficacious than its initiation after the clinical manifestation of diabetes. However, the appropriate immunotherapeutical strategies are yet to be worked out.


On the crystallisation temperature of very high-density amorphous ice.
The influence of the protocol of preparation on the crystallisation temperature TX of very high-density amorphous ice (VHDA) was studied by varying the annealing pressure (1.1, 1.6 and 1.9 GPa) and temperature (160, 167 and 175 K, respectively). TX increases by up to 4 K in the pressure range of 0.7 to 1.8 GPa for samples annealed at 1.9 GPa compared to samples annealed at 1.1 GPa. Concomitantly, secondary crystallisation channels are suppressed, indicating the absence of structural inhomogeneities. For VHDA prepared at 1.1 GPa and 1.6 GPa our results indicate such inhomogeneities, which we regard to be incompletely amorphized, distorted nanodomains of hexagonal ice that cannot be detected through X-ray diffraction experiments. VHDA prepared at high pressures and temperatures thus represents the amorphous state of water at >0.7 GPa least affected by nanocrystals that has been described so far. We expect the TX obtained for the samples prepared in this manner to be close to the ultimate limit, i.e., we do not consider it possible to raise the low-temperature border to the no-man's land notably further by changing the preparation protocol. An additional, considerable increase in this border will only be possible by working at much shorter time-scales, e.g., by employing fast heating experiments.


[Multimodal treatment of colon cancer?].
Surgical therapy is still the basis of therapy of patients with colon carcinoma. Multimodal therapeutical concepts are presently applied as a therapeutical standard in the adjuvant therapy and increasingly in the systemic therapy of patients with primarily inoperable metastases of the liver to reach a secondary operability. Interdisciplinary multimodal therapeutical concepts are even accepted within the therapy of metastasized colon carcinomas. There are still unanswered questions regarding sequences of palliative systemic therapies and their combinations with local ablative methods.


[Quantification of left ventricular function and mass in dual-source CT (DSCT).].
To evaluate the interobserver agreement in quantifying left ventricular function and mass and to assess the accuracy of conventional manual contour tracing compared to semiautomatic segmentation analysis software. Twenty consecutive subjects who underwent cardiac DSCT with retrospective ECG-gating were included. Two different multiphase image reconstructions were done in 5% steps throughout the entire cardiac cycle (0-95% of the R-R interval) with effective slice thickness of 1mm in the axial plane and 8mm in the short-axis orientation. Left ventricular function and mass were calculated by two independent observers, tracing endocardial and epicardial borders manually and using a semiautomatic software tool (Circulation II, Siemens). Ejection fraction (EF), end-diastolic volume (EDV), end-systolic volume (ESV), stroke volume (SV), cardiac output (CO), and myocardial mass were evaluated by two independent observers blind to each other's assessments. The interobserver agreement and the reliability of the different segmentation methods were calculated. The time required for manual contouring and semiautomatic contour tracing was also registered. We found an excellent correlation (r>0.94; p<0.05) between the two independent observers for the quantification of left ventricular function and mass. Left ventricular functional parameters derived from semiautomatic contour software and conventional manual tracing method were not significantly different (p>0.05). The semiautomatic contour detection algorithm overestimated LV mass significantly compared with the manual contouring method (mean difference 29.45+/-1.64g; p<0.05). The time needed to calculate these parameters with the semiautomatic tool was significantly lower (248.85+/-99.8s) than with manual contouring (452.7+/-73.92s) (p<0.05). Interobserver agreement for quantifying left ventricular function and mass using DSCT is excellent. Despite overestimating left ventricular mass, the semiautomatic software tool allows cardíac parameters to be quantified with the same reliability as the conventional manual method in half the time.