Electrochemical Oxidation of Δ9-Tetrahydrocannabinol at Nanomolar Concentrations.
With increasing marijuana legalization, there is a growing need for technology that can determine if an individual is impaired due to recent marijuana usage. The electrochemical oxidation of Δ9-THC to form its corresponding quinones can be used as a framework to develop an electrochemical sensor for Δ9-THC. This study describes an electrochemical oxidation of Δ9-THC that uses a copper anode, a platinum cathode, and an atmosphere of oxygen. The oxidation is feasible at nanomolar concentrations, which approaches the reactivity that is necessary for developing a real-world marijuana breathalyzer. Moreover, we show that vaporized Δ9-THC can be captured directly in an electrolyte medium and subjected to electrochemical oxidation, thus paving the way for use in future technology development.


Disruption of conserved polar interactions causes a sequential release of Bim mutants from the canonical binding groove of Mcl1.
Mcl1 is an important anti-apoptotic member of the Bcl2 family proteins that are upregulated in several cancer malignancies. The canonical binding groove (CBG) located at the surface of Mcl1 exhibits a critical role in binding partners selectively via the BH3-domain of pro-apoptotic Bcl2 family members that trigger the downregulation of Mcl1 function. There are several crystal structures of point-mutated pro-apoptotic Bim peptides in complex with Mcl1. However, the mechanistic effects of such point-mutations towards peptide binding and complex stability still remain unexplored. Here, the effects of the reported point mutations in Bim peptides and their binding mechanisms to Mcl1 were computationally evaluated using atomistic-level steered molecular dynamics (SMD) simulations. A range of external-forces and constant-velocities were applied to the Bim peptides to uncover the mechanistic basis of peptide dissociation from the CBG of Mcl1. Although the peptides showed similarities in their dissociation pathways, the peak rupture forces varied significantly. According to simulations results, the disruption of the conserved polar contacts at the complex interface causes a sequential release of the peptides from the CBG of Mcl1. Overall, the results obtained from the current study may provide valuable insights for the development of novel anti-cancer peptide-inhibitors that can downregulate Mcl1's function.


The effects of respiratory training for chronic obstructive pulmonary disease patients: a randomised clinical trial.
To assess the effects of respiratory training on lung function, activity tolerance and quality of life in patients with chronic obstructive pulmonary disease. For patients with chronic obstructive pulmonary disease, pulmonary rehabilitation can increase activity tolerance and improve their psychological state by relieving dyspnoea and promoting their quality of life. A randomised clinical trial was conducted in a local hospital. Patients with chronic obstructive pulmonary disease were randomly assigned to intervention (n=20) and control groups (n=20). Spirometry, six-minute walking distance and quality of life were used to assess the efficacy of respiratory training programme. Significant improvement in lung function, including forced vital capacity (p=0.037), forced expiratory volume in one-second (p=0.006) and per cent predicted forced expiratory volume in one-second (p=0.008) in the intervention group. Regarding efficacy of the training programme for patients with chronic obstructive pulmonary disease, in forced expiratory volume in one-second (p=0.024) and per cent predicted forced expiratory volume in one-second (p=0.035), six-minute walking distance significantly increased. In addition, there were significant improvements for symptoms (p=0.018), impact (p<0.001) and total quality of life scores (p<0.001), as well as significantly decreased body mass, airflow obstruction, dyspnoea and exercise capacity index (p=0.004) in the intervention group. A respiratory training programme for patients with chronic obstructive pulmonary disease was found to relieve dyspnoea, maintain lung function, increase activity tolerance and improve quality of life. Respiratory training programme can be used as a routine rehabilitation protocol for patients with chronic obstructive pulmonary disease and can be used by nurses as a reference to monitor chronic obstructive pulmonary disease patients' health status.


Novel insights in the genomic organization and hotspots of recombination in the human KIR locus through analysis of intergenic regions.
The Killer Immunoglobulin-like Receptor (KIR) proteins constitute a family of highly homologous surface receptors involved in the regulation of the innate cytotoxicity of natural killer (NK) cells. Within the human genome, 17 KIR genes are present, many of which show large variation across the population owing to the high number of allelic variants and copy number variation (CNV). KIR genotyping and CNV determination were used to map the KIR locus in a large cohort of >400 Caucasian individuals. Gene order and structure was determined by sequence-specific polymerase chain reaction of the intergenic regions. In this way, we could show that KIR3DL1 and KIR2DS4 gene variants are linked and that--contrary to current views--the gene KIR2DS5 is only present in the telomeric half of the KIR locus. Our study revealed novel insights in the highly organized distribution of KIR genes. Novel recombination hotspots were identified that contribute to the diversity of KIR gene distribution in the Caucasian population. Next-generation sequencing of the KIR intergenic regions allowed for a detailed single-nucleotide polymorphism analysis, which demonstrated several gene-specific as well as haplotype-specific nucleotides for a more accurate genotyping of this notoriously complex gene cluster.


beta 2-Microglobulin and its relationship to the immune system.
beta 2-Microglobulin has been isolated from several species, but only bovine beta 2-microglobulin, previously known as lactollin, has been crystallized. An improved method for its isolation from colostrum is described. The bovine homologue exhibits a concentration-dependent aggregation behavior. beta 2-Microglobulin is related to both immune and histocompatibility antigen systems. It exhibits homology with the constant domains of the immunoglobulin-G light and heavy chains and is an integral part of histocompatibility antigens bound to cell surface. beta 2-Microglobulin also occurs in the free state in various body fluids including milk and colostrum. The possible relationship of elevated free beta 2-microglobulin of pathological conditions is suggested for future research.


Recent gene conversion between genes encoding human red and green visual pigments.
Nucleotide sequences of the human X-linked red and green pigment genes were compared, and the number of silent substitutions per site (KSc) between these genes was analysed in comparison with the corresponding values of primate genes. Taking the retarded mutation rate of X-linked genes into consideration (Miyata et al., 1987), the red and green pigment genes were shown to have undergone gene conversion at around the time of separation of African apes and orangutan. Thus the recent gene conversion and retarded mutation rate in these X-linked genes are probably responsible for the strong sequence similarity between these genes, which is likely to facilitate the occurrence of red-green color blindness in the human population. It was also shown that the red pigment gene evolved about five times more rapidly than the green pigment gene since the latest gene conversion.


Synthesis and characterization of magnesium oxide supported catalysts with a meso-macropore structure.
Nanostructured magnesium oxide catalyst support materials with controlled morphology and size were synthesized from a supercritical carbon dioxide/ethanol solution via chemical reaction of soluble precursors and subsequent thermal decomposition. Leaf-like magnesium hydroxide precursors with high specific surface area were formed by a low-temperature chemical reaction at below 140 degrees C, while magnesium carbonate cubes with a very low surface area less than 3.3 m2/g were formed by temperature-induced phase transition from a loose to a dense structure during carbonation reaction at above 150 degrees C. The specific surface area has significantly increased higher than 90 m2/g due to the creation of highly porous MgO cubes with mesopore structure formed after thermal decomposition of the magnesium carbonate precursors. Ni-magnesium oxide catalysts with bimodal pore structure were finally formed by the consequence of packing heterogeneity of the porous magnesium oxides with different morphologies and sizes.


A Self-Healable Bifunctional Electronic Skin.
For the mimicry of human skin, one of the challenges is how to detect and recognize different stimulus by electronic device, while still has the ability of skin self-recovery at the same time. Because of the excellent elasticity and flexibility, strong self-healing ability, in this paper, we reported a bifunctional self-healing e-skin with polyurethane (PU) and polyurethane@multiwalled carbon nanotubes (PU@CNT) as the sensing materials by integrating a resistance temperature sensor on top of a capacitive pressure sensor on the same flexible cellulose nanocrystals@carboxylated nitrile rubber@polyethylenimine (CNC@XNBR) substrate. Studies found that each type of sensor exhibited fast and superior response to only the target stimuli. Meanwhile, due to the self-recovery properties of PU and CNC@XNBR, as-fabricated e-skin has the self-healing ability after damage and remains excellent sensitivity to temperature and pressure after healing. A 5 × 5 device array was also fabricated, which can simultaneously image the pressure and temperature distribution.


Remodeling arteries: studying the mechanical properties of 3D-bioprinted hybrid photoresponsive materials.
3D-printed cell models are currently in the spotlight of medical research. Whilst significant advances have been made, there are still aspects that require attention to achieve more realistic models which faithfully represent the in vivo environment. In this work we describe the production of an artery model with cyclic expansive properties, capable of mimicking the different physical forces and stress factors that cells experience in physiological conditions. The artery wall components are reproduced using 3D printing of thermoresponsive polymers with inorganic nanoparticles (NPs) representing the outer tunica adventitia, smooth muscle cells embedded in extracellular matrix representing the tunica media, and finally a monolayer of endothelial cells as the tunica intima. Cyclic expansion can be induced thanks to the inclusion of photo-responsive plasmonic NPs embedded within the thermoresponsive ink composition, resulting in changes in the thermoresponsive polymer hydration state and hence volume, in a stimulated on-off manner. By changing the thermoresponsive polymer composition, the transition temperature and pulsatility can be efficiently tuned. We show the direct effect of cyclic expansion and contraction on the overlying cell layers by analyzing transcriptional changes in mechanoresponsive mesenchymal genes associated with such microenvironmental physical cues. The technique described herein involving stimuli-responsive 3D printed tissue constructs, also described as four- dimensional (4D) printing, offers a novel approach for the production of dynamic biomodels.


Krukenberg tumors of the ovary: a clinicopathologic analysis of 112 cases.
The records from 1965 to 1985 in the Department of Obstetrics and Gynecology Kurume University Hospital were reviewed. In this period, 112 cases were fulfilled the criteria of Krukenberg tumor and they were carefully screened to clinical history, physical findings, operative findings, and histologic character of the lesion. The incidence of Krukenberg tumors in all ovarian cancer was 17.8%. The average age was 45.4 years with ranged from 23 to 71. As expected from this age distribution, 73 patients (65%) were 50 years of age or younger. A primary carcinoma of the stomach was found in 78 (69.6%) of 112 patients. In three cases, the Krukenberg tumors were associated with a recent pregnancy. Two cases were associated with ovarian dermoid cyst. Microscopically, both ovaries were involved in all cases that could be evaluated for bilaterality. The relation between age distribution and histological characteristics was examined. Signet-ring type carcinomas were observed frequently in young women and gland-formation type carcinomas in older women. Stromal hyperplasia of the tumors were remarkably common in younger women. In the patients with a recent pregnancy, the abdominal mass developed rapidly after delivery. In addition, estrogen receptors were present in all experimented cases. The above results suggested that estrogen influences tumor development and the histological character many of Krukenberg tumor.


The transcription factors GATA4 and dHAND physically interact to synergistically activate cardiac gene expression through a p300-dependent mechanism.
An intricate array of heterogeneous transcription factors participate in programming tissue-specific gene expression through combinatorial interactions that are unique to a given cell-type. The zinc finger-containing transcription factor GATA4, which is widely expressed in mesodermal and endodermal derived tissues, is thought to regulate cardiac myocyte-specific gene expression through combinatorial interactions with other semi-restricted transcription factors such as myocyte enhancer factor 2, nuclear factor of activated T-cells, serum response factor, and Nkx2.5. Here we determined that GATA4 also interacts with the cardiac-expressed basic helix-loop-helix transcription factor dHAND (also known as HAND2). GATA4 and dHAND synergistically activated expression of cardiac-specific promoters from the atrial natriuretic factor gene, the b-type natriuretic peptide gene, and the alpha-myosin heavy chain gene. Using artificial reporter constructs this functional synergy was shown to be GATA site-dependent, but E-box site-independent. A mechanism for the transcriptional synergy was suggested by the observation that the bHLH domain of dHAND physically interacted with the C-terminal zinc finger domain of GATA4 forming a higher order complex. This transcriptional synergy observed between GATA4 and dHAND was associated with p300 recruitment, but not with alterations in DNA binding activity of either factor. Moreover, the bHLH domain of dHAND directly interacted with the CH3 domain of p300 suggesting the existence of a higher order complex between GATA4, dHAND, and p300. Taken together with previous observations, these results suggest the existence of an enhanceosome complex comprised of p300 and multiple semi-restricted transcription factors that together specify tissue-specific gene expression in the heart.


The global structure of the VS ribozyme.
The VS ribozyme comprises five helical segments (II-VI) in a formal H shape, organized by two three-way junctions. It interacts with its stem-loop substrate (I) by tertiary interactions. We have determined the global shape of the 3-4-5 junction (relating helices III-V) by electrophoresis and FRET. Estimation of the dihedral angle between helices II and V electrophoretically has allowed us to build a model for the global structure of the complete ribozyme. We propose that the substrate is docked into a cleft between helices II and VI, with its loop making a tertiary interaction with that of helix V. This is consistent with the dependence of activity on the length of helix III. The scissile phosphate is well placed to interact with the probable active site of the ribozyme, the loop containing A730.


Comparison of coronary hemodynamics during infusions of dobutamine and adenosine in patients with angina pectoris.
Pharmacologic stress tests are used to evaluate the significance of coronary arterial stenosis, and adenosine and dobutamine are often used for this purpose. The aim of this study was to examine coronary hemodynamics in 16 patients with stable exertional angina who had organic stenosis (> 90%) in the left anterior descending coronary arteries and in 13 control subjects at baseline and during separate intravenous infusions of dobutamine (20 micrograms/kg per min [corrected]) and adenosine (140 micrograms/kg per min [corrected]). Measurements of great cardiac vein blood flow and lactate and oxygen contents in the blood from the aortic root and great cardiac vein were analyzed in patients with ischemic signs of myocardial ischemia (ST-segment depression in the precordial ECG leads and/or myocardial lactate production) during the infusions (in 11 patients during adenosine infusion and in 12 patients during dobutamine infusion) and in control subjects. Myocardial oxygen consumption during adenosine infusion did not change compared with the baseline in both groups, whereas it increased during dobutamine infusion in both groups (p < 0.01 compared with the baseline in each case). However, great cardiac vein blood flow was significantly increased compared with the baseline during both infusions in both groups (p < 0.01 compared with baseline in each case), and the oxygen contents of great cardiac vein blood, which contains effluent from the ischemic region, showed no decrease but increased significantly during both infusions in the patients in comparison with control subjects. Although the global oxygen supply was excessive, myocardial ischemia occurred during dobutamine and adenosine infusions in the patients. As myocardial oxygen consumption remained unchanged during adenosine infusion, regional myocardial flow reduction, possibly as a result of flow maldistribution, may contribute, at least partly, to adenosine-induced myocardial ischemia. An increase in myocardial oxygen consumption may be the principal cause of myocardial ischemia during dobutamine infusion, but an inappropriate flow increase in normal myocardium may occur during dobutamine-induced ischemia.


[Transabdominal removal of femoral and acetabular components of a severely protruded and infected hip arthroplasty with urinary tract complications].
We present the first report of transabdominal removal of femoral and acetabular components of a severely loosened hip prosthesis protruding into the pelvis. In a 73-year-old woman post-operative development of urinary tract complications emphasize importance of careful assessment of the prosthetic relations with the vascular and nervous structures as well as pelvic organs before removal of the hip prosthesis. Angio-computed tomography is the most contributive exploration to assess vascular relations. In patients with particular clinical presentations or with threatened structures in the vicinity of the prosthesis, this examination must be completed by complementary opacifications (urinary and gastrointestinal tracts, joints). Ureteral catheterization may be needed if the structures are close or if there is a suspected modification of the urinary tract (retraction, mass effect). In present case, we did not opacify the urinary tract before laparatomy despite the presence of urinary signs preoperatively. A suspected ureterovaginal fistula was discovered. But they where also a ureteral lesions which can result from difficult dissection in contact with infected tissues. In this patient, urinary complications led to nephrectomy after temporary pyelostomy for urine bypass. At last follow-up, the urinary tract infection was controlled but reimplantation was not attempted because of insulin dependent diabetes mellitus and poor general condition. The spontaneous course of this infection with prosthesis loosening recalls the importance of regular surveillance of total hip replacements.


Lamin B1 fluctuations have differential effects on cellular proliferation and senescence.
The nuclear lamina consists of A- and B-type lamins. Mutations in LMNA cause many human diseases, including progeria, a premature aging syndrome, whereas LMNB1 duplication causes adult-onset autosomal dominant leukodystrophy (ADLD). LMNB1 is reduced in cells from progeria patients, but the significance of this reduction is unclear. In this paper, we show that LMNB1 protein levels decline in senescent human dermal fibroblasts and keratinocytes, mediated by reduced transcription and inhibition of LMNB1 messenger ribonucleic acid (RNA) translation by miRNA-23a. This reduction is also observed in chronologically aged human skin tissue. To determine whether altered LMNB1 levels cause senescence, we either increased or reduced LMNB1. Both LMNB1 depletion and overexpression inhibited proliferation, but only LMNB1 overexpression induced senescence, which was prevented by telomerase expression or inactivation of p53. This phenotype was exacerbated by a simultaneous reduction of LMNA/C. Our results demonstrate that altering LMNB1 levels inhibits proliferation and are relevant to understanding the molecular pathology of ADLD.


Role of microparticles in endothelial dysfunction and arterial hypertension.
Microparticles are small cell vesicles that can be released by almost all eukaryotic cells during cellular stress and cell activation. Within the last 1-2 decades it has been shown that microparticles are useful blood surrogate markers for different pathological conditions, such as vascular inflammation, coagulation and tumour diseases. Several studies have investigated the abundance of microparticles of different cellular origins in multiple cardiovascular diseases. It thereby has been shown that microparticles released by platelets, leukocytes and endothelial cells can be found in conditions of endothelial dysfunction, acute and chronic vascular inflammation and hypercoagulation. In addition to their function as surrogate markers, several studies indicate that circulating microparticles can fuse with distinct target cells, such as endothelial cells or leukocyte, and thereby deliver cellular components of their parental cells to the target cells. Hence, microparticles are a novel entity of circulating, paracrine, biological vectors which can influence the phenotype, the function and presumably even the transcriptome of their target cells. This review article aims to give a brief overview about the microparticle biology with a focus on endothelial activation and arterial hypertension. More detailed information about the role of microparticles in pathophysiology and disease can be found in already published work.


Acquired cystic kidney disease following long-term peritoneal dialysis for congenital nephrotic syndrome.
We describe here the clinicopathological findings in a child with congenital nephrotic syndrome (CNS) non-responsive to medical therapy who developed acquired cystic kidney disease (ACKD) in both native kidneys after long-term peritoneal dialysis. This case indicates that CNS is a further pathologic condition related to the development of ACKD.


3D spongy nanofiber structure Fe-NC catalysts built by a graphene regulated electrospinning method.
3D spongy nanofiber structure Fe-NC catalysts were constructed by a graphene regulated electrospinning method. The framework of the catalysts was reconstructed into carbon nanotubes, mesopores and macropores, and most of the Fe3C is converted to Fe2N during the calcination process. All catalysts showed better electrocatalytic performances than commercial Pt/C.


Immune and genetic therapies for advanced renal cell carcinoma.
Although we have witnessed advances in many aspects of cancer research and therapy in recent years, the ability to cure the majority of patients with advanced renal cell carcinoma (RCC) remains elusive. At the same time, it has become increasingly apparent that a better understanding of the genetic alterations and immune dysregulations in RCC will play a key role in finding a treatment. Therefore, clinical trials directed at specific genetic alterations and studies exploiting components of the immune system are being conducted. These studies provide new hope for an improved outlook for patients presenting with advanced RCC. The future prospects of RCC therapy will be, without doubt, built on the foundation of current investigative efforts in gene and immune therapy. This article reviews the current role of immunotherapy and gene therapy in the management of metastatic RCC. Finally, current clinical trials focusing on gene and immune therapies are listed.


Catecholamine-induced myocardial potassium uptake mediated by beta 1-adrenoceptors and adenylate cyclase activation in the pig.
The myocardial potassium uptake during intracoronary isoproterenol stimulation was characterized in 12 anesthetized pigs. The beta-receptor subtype specificity and the effect of adenylate cyclase activation were determined. Potassium concentrations were continuously recorded by PVC-valinomycin minielectrodes in the left atrial cavity and in coronary sinus blood diverted through a shunt to the right atrium. The difference in potassium concentration between the left atrial cavity and coronary sinus, and the accumulated myocardial potassium uptake were calculated after computerized data sampling. By intracoronary drug infusion, changes in heart rate and systemic effects were minimized. Isoproterenol (0.6-0.8 microgram/min), a nonspecific beta-agonist, reduced coronary sinus potassium concentration transiently to a nadir of 0.28 (0.15-0.43) mM (median and 95% confidence interval) below control values (n = 12). The potassium uptake, which amounted to 140 (79-202) mumol/100 g tissue, corresponding to an intracellular potassium increase of about 3 mM, was abolished after selective beta 1-blockade by pafenolol. The specific beta 1-agonist dobutamine (40 micrograms/min) caused a similar potassium uptake before and after selective beta 2-blockade by ICI 118, 551. Salbutamol (2 micrograms/min), a specific beta 2-agonist, induced a minor potassium uptake of 4 (1-20) mumol/100 g, blocked by pafenolol. After nonselective beta-blockade by propranolol the adenylate cyclase stimulator forskolin caused a myocardial potassium uptake of similar magnitude to that of isoproterenol before beta-blockade. We conclude that a myocardial potassium uptake ensues during beta 1-adrenoceptor stimulation and adenylate cyclase activation.


Towards improved diagnosis of von Willebrand disease: comparative evaluations of several automated von Willebrand factor antigen and activity assays.
von Willebrand disease (VWD) is reportedly the most common bleeding disorder and arises from deficiency and/or defects of von Willebrand factor (VWF). Laboratory diagnosis and typing has important management implications and requires a wide range of tests, including VWF activity and antigen, and involves differential identification of qualitative vs quantitative defects. We have assessed several VWF antigen and activity assays (collagen binding [VWF:CB], ristocetin cofactor [VWF:RCo] and the new Siemens INNOVANCE assay [VWF:Ac], employing latex particles and gain of function recombinant glycoprotein Ib to facilitate VWF binding and agglutination without need for ristocetin) using different instrumentation, including the new Sysmex CS-5100, with a large sample test set (n=600). We included retrospective plus prospective study designs, and also evaluated desmopressin responsiveness plus differential sensitivity to high molecular weight VWF. VWF:Ag and VWF:RCo results from different methods were respectively largely comparable, although some notable differences were evident, including one high false normal VWF:Ag value (105 U/dL) on a type 3 VWD sample, possibly due to heterophile antibody interference in the latex-based CS-5100 methodology. VWF:Ac was largely comparable to VWF:RCo, but VWF:CB showed discrepant findings to both VWF:RCo and VWF:Ac with some patients, most notably patients with type 2M VWD. (a) VWF:Ag on different platforms are largely interchangeable, as are VWF:RCo on different platforms, except for occasional (some potentially important) differences, and manufacturer recommended methods may otherwise require some assay optimization; (b) VWF:RCo and VWF:Ac are largely interchangeable, except for occasional differences that may also relate to assay design (differing optimizations); (c) VWF:CB provides an additional activity to supplement VWF:RCo or VWF:Ac activity assays, and is not interchangeable with either.


[Electron microscopic observations on the extrusion of enteroendocrine cells from the epithelium of the mouse small intestine].
By means of electronmicrographs it could be showed that enteroendocrine cells at the tips of the villi intestinalis are extruded in a way similar to other intestinal cells. They lose their contact to the basal lamina. An at first small process protrudes into the intestinal lumen. The cell as an entity can follow afterwards. Frequently this extrusion takes place together with neighbouring cells and as a consequence, there is a cell complex in the intestinal lumen which can be connected with the epithelium by means of a small cytoplasm bridge. A second way leads over rupturing of the protruding part of the cell and the extrusion of the cell residue.


Complete genome sequences of three ebola virus isolates from the 2014 outbreak in west Africa.
Here, we report the complete genome sequences, including the genome termini, of three Ebola virus isolates (species Zaire ebolavirus) originating from Guinea that are now being widely used in laboratories in North America for research regarding West African Ebola viruses.


Perpetuating stigma? Differences between advertisements for psychiatric and non-psychiatric medication in two professional journals.
Continuing debates regarding advertising and the pharmaceutical industry, and others detailing the continued stigmatization of mental health problems. To establish whether there are any differences in advertisements for psychiatric and non-psychiatric medication aimed at health professionals. Quantitative (t-tests, Chi-squared) and qualitative analysis of all unique advertisements for medication that appeared in two professional journals (the British Medical Journal and the British Journal of Psychiatry) between October 2005 and September 2006 was undertaken. Close attention was paid to both images and text used in the advertisements. Significant differences were found between advertisements for psychiatric and non-psychiatric medication in both quantitative and qualitative analysis: advertisements for psychiatric medication contain less text and are less likely to include specific information about the actual drug than non-psychiatric medication advertisements; images used in advertisements for psychiatric medication are more negative than those used for non-psychiatric medication, and are less likely to portray people in everyday situations. A distinction between mental health problems and other forms of ill health is clearly being maintained in medication advertisements; this has potentially stigmatizing consequences, both for professional and public perceptions. There are also troubling implications in light of the debates surrounding Direct to Consumer Advertising.


Microbiology of infection in mechanical circulatory support.
The results of MCSS therapy for patients with severe heart failure are improving, and there are increasing numbers of implants as device therapy enters the era of permanent use (i.e., destination therapy). Device-related infection of implanted pumps and sepsis remain important risk factors for death, and once infections are established on biomaterial surfaces they usually persist despite prolonged antimicrobial therapy. This paper covers the topic of infection in mechanical circulatory support. Methods to prevent and manage infection are described. Recent information from the field of microbiology that is relevant to understanding device-related infection is reviewed, including the mechanisms for microbial adhesion to prosthetic surfaces, quorum sensing, and biofilm formation. Characteristics that are important to antimicrobial resistance of microbes dwelling in biofilms are described (e.g. responses to microenvironmental conditions that result in resistance to antibiotics). This information may lead to new therapies that further diminish rates of device infection, and allow complete eradication of infections when they occur.


The Effect of a Compound Protein on Wound Healing and Nutritional Status.
Proteins provide the main building blocks for tissue growth, cell renewal, and repair during wound healing. We aimed to examine the effect of a compound protein on wound healing, nutritional status, and underlying mechanisms. We first performed a preliminary experiment to identify the appropriate wound healing assessment points. In the formal experiment, there were five groups (control group: 8.3750 g/kg/day saline solution; model group: 8.3750 g/kg/day saline solution; whey protein group: 8.3750 g/kg/day whey protein; low-dose compound protein group: 4.1875 g/kg/day compound protein; and high-dose compound protein group: 8.3750 g/kg/day compound protein) with eight rats in each group. At each turning point, we observed the wound healing rate and nutritional status of the different groups of rats. In addition, biochemical assays were used to determine the mechanisms underlying the effects of the compound protein. In the preliminary experiment, the third day after modeling was the turning point between the inflammatory and proliferation phases, and the eighth day was the turning point between the proliferation and remodeling phases. The formal experiment evaluated wound healing condition, inflammatory response, angiogenesis, collagen deposition, and nutritional status. A pathological report showed increased vascularization, collagen deposition, and epithelialization in compound protein-treated groups. Protein-treated mice showed decreased interleukin (IL)-6, IL-8, neutrophils, and lymphocytes and increased IL-10, albumin, prealbumin, total protein levels, insulin-like growth factor 1 (IGF-1), fibroblast growth factor 2 (FGF-2), and vascular endothelial growth factor (VEGF) expressions. All parameters were significant (p < 0.05) compared to the model group. There was a dose-dependent effect of the compound protein. The accelerated wound healing mechanism may be that the compound protein accelerates the whole wound healing process, making wounds transition from the inflammatory phase to the proliferation phase faster, entering the remodeling phase earlier. Administration of a compound protein can accelerate wound healing and improve the nutritional status.


The efficacy of a video-based marker-less tracking system for gait analysis.
Most clinical gait analyses are conducted using motion capture systems which track retro-reflective markers that are placed on key landmarks of the participants. An alternative to a three-dimensional (3D) motion capture, marker-based, optical camera system may be a marker-less video-based tracking system. The aim of our study was to investigate the efficacy of the use of a marker-less tracking system in the calculation of 3D joint angles for possible use in clinical gait analysis. Ten participants walked and jogged on a treadmill and their kinematic data were captured with a marker and marker-less tracking system simultaneously. The hip, knee and ankle angles in the frontal, sagittal and transverse planes were computed. Root Mean Square differences (RMSdiff) between corresponding angles for each participant's support phase were calculated and averaged to derive the mean within-subject RMSdiff. These within-subject means were averaged to obtain the mean between-subject RMSdiff for the relevant joint angles in the two gait conditions (walking and jogging). The RMSdiff between the two tracking systems was less than 1° for all rotations of the three joint angles of the hip and knee. However, there were slightly larger differences in the ankle joint angles. The results of this study suggest a potential application in gait analysis in clinical settings where observations of anatomical motions may provide meaningful feedback.


Zero GVD slow-light originating from a strong coupling of one-way modes in double-channel magneto-optical photonic crystal waveguides.
We have studied the coupling effect of topological photonic states in a double-channel magneto-optical photonic crystal waveguide by introducing a two-stranded ordinary Al2O3 photonic crystal as the coupling layer. There exist both M1 (odd) and M2 (even) one-way modes simultaneously in the bandgap. Interestingly, M1 mode is always a fast-light mode with large group velocity (vg) and large group velocity dispersion (GVD) regardless what the radius (RA) of Al2O3 rods is. However, when RA is appropriate, M2 mode becomes a very slow-light mode exhibiting near-zero vg and zero GVD simultaneously. The physical reason of such slow-light is attributed to the strong coupling effect between the one-way edge modes in both sub-waveguides. Furthermore, the simulation results show that the robustness of both the fast- and slow-light modes are extremely strong against perfect electric conductor defect and the one-way transmittance is close to 100%. Besides, the PEC defect can cause significant phase delay. These results hold promise for many fields such as signal processing, optical modulation, and the design of various topological devices.


Transport properties in a Sb-Te binary topological-insulator system.
Sb-Te layers having various compositions between Sb2Te3 and Sb2Te are grown using molecular beam epitaxy. The structural and electrical properties of the layers change gradually with composition but exhibit a discontinuity involving a bistability. The holes in the layers are generated by Sb bilayers intercalated between Sb2Te3 quintuple layers and their mobility is governed by the scattering from the parent acceptors. Magnetoresistance for compositions around SbTe is linear, for which the reduction of the parabolic component due to low mobility is crucial. Density functional calculations predict Sb2Te3 and SbTe to be topological insulators (TIs) resembling Bi2Se3 and Bi2Te3, respectively. The prefactor of the weak antilocalization effect is α =- 1 regardless of the composition. The Sb-Te system is thus a family of TIs possessing undisturbed surface states for which the location of the Dirac point with respect to the bulk band gap is adjustable.


Peliosis Hepatis Complicating Pregnancy: A Rare Entity.
Peliosis hepatis (PH) is a rare, benign condition of the liver characterized by the presence of blood-filled lacunar spaces in the parenchyma. It usually has a chronic presentation and is a rare cause of portal hypertension reported in adult patients. Its etiology is diverse and ranges from infectious agents to tumors to toxic substances and anabolic steroids; however, the cause remains unclear in 25-50% of patients. Similarly, the symptomatology and imaging findings are diverse. Biopsy is the definitive test to diagnose the condition. Herein, we present a case of a young female presenting in her seventh month of gestational amenorrhea with signs of portal hypertension and subsequently diagnosed to have PH. She was managed conservatively and delivered her baby normally. Later, she presented with spontaneous bacterial peritonitis and hepatic encephalopathy and developed hepatorenal syndrome. She later succumbed to her illness. The condition should be kept in the differential diagnosis of the atypical liver masses and liver diseases causing portal hypertension.


Combination of rapamycin and garlic-derived S-allylmercaptocysteine induces colon cancer cell apoptosis and suppresses tumor growth in xenograft nude mice through autophagy/p62/Nrf2 pathway.
The natural plant-derived product S-allylmercapto-cysteine (SAMC) has been studied in cancer therapy as a single and combination chemotherapeutic agent. The present study was employed to verify the combination use of SAMC and rapamycin that is the mTOR inhibitor with anticancer ability but has limited efficacy due to drug resistance, and to explore the underlying mechanisms. We combined rapamycin and SAMC for colorectal cancer treatment in the HCT‑116 cancer cells and a xenograft murine model. The in vivo study was established by xenografting HCT‑116 cells in BALB/c nude mice. It was found that the combination therapy had enhanced tumor-suppressing ability with the upregulation of the Bax/Bcl-2 ratio as a consequence of activated apoptosis, inhibition of autophagic activity and prevention of Akt phosphorylation. The rapamycin and SAMC combination activated antioxidant transcription expressions of Nrf2 and downstream gene NQO1. Concomitantly, autophagosome cargo p62 was downregulated, indicating that the p62 played a negative-regulatory role between Nrf2 and autophagy. Our results show that the combination of SAMC and rapamycin enhanced the anticancer ability, which could be used for the treatment of colorectal cancer. The underling mechanism of autophagy/p62/Nrf2 pathway discovered may provide a new direction for drug development, especially for traditional Chinese medicines.


Prostacyclin reverses platelet stress fibre formation causing platelet aggregate instability.
Prostacyclin (PGI2) modulates platelet activation to regulate haemostasis. Evidence has emerged to suggest that thrombi are dynamic structures with distinct areas of differing platelet activation. It was hypothesised that PGI2 could reverse platelet spreading by actin cytoskeletal modulation, leading to reduced capability of platelet aggregates to withstand a high shear environment. Our data demonstrates that post-flow of PGI2 over activated and spread platelets on fibrinogen, identified a significant reduction in platelet surface area under high shear. Exploration of the molecular mechanisms underpinning this effect revealed that PGI2 reversed stress fibre formation in adherent platelets, reduced platelet spreading, whilst simultaneously promoting actin nodule formation. The effects of PGI2 on stress fibres were mimicked by the adenylyl cyclase activator forskolin and prevented by inhibitors of protein kinase A (PKA). Stress fibre formation is a RhoA dependent process and we found that treatment of adherent platelets with PGI2 caused inhibitory phosphorylation of RhoA, reduced RhoA GTP-loading and reversal of myosin light chain phosphorylation. Phospho-RhoA was localised in actin nodules with PKA type II and a number of other phosphorylated PKA substrates. This study demonstrates that PGI2 can reverse key platelet functions after their initial activation and identifies a novel mechanism for controlling thrombosis.


Post-myocardial infarction ventricular remodeling: animal and human studies.
Segmental alterations in left ventricular function are generally present in patients who suffer an acute myocardial infarction. Regional wall motion abnormalities in left ventricular systolic function can be identified in the hyperacute period and generally persist in patients who complete a myocardial infarction. Through the process of infarct expansion, the infarcted territory may thin and lengthen in the short term following a myocardial infarction. Some infarct survivors are also prone to further progressive alterations in the shape and size of the left ventricle, a process that has been termed postinfarction ventricular remodeling. Although left ventricular remodeling appears to represent an adaptive process serving to preserve stroke volume (and cardiac output) following myocardial injury, the enlargement process may have undesirable long-term effects on global left ventricular function and on clinical prognosis. Fortunately, recent experimental and clinical evidence demonstrates that ventricular remodeling and its deleterious consequences may be preventable.


Inferior Vena Cava Thrombus due to Left Inferior Vena Cava and Ulcerative Colitis.
A 29-year-old man with diarrhea and abdominal pain for 2 weeks presented with new-onset left back pain. Contrast-enhanced computed tomography (CT) showed a left inferior vena cava (IVC) crossing over the aorta, and thrombus in the IVC and left renal vein. Colonoscopy and biopsy for assessment of diarrhea and abdominal pain provided a diagnosis of ulcerative colitis. Stasis of blood flow due to left IVC crossing over the aorta, and hypercoagulability due to ulcerative colitis influenced thrombus formation.


The Visible Human Project: From Body to Bits.
Atlases of anatomy have long been a mainstay for visualizing and identifying features of the human body [1]. Many are constructed of idealized illustrations rendered so that structures are presented as three-dimensional (3-D) pictures. Others have employed photographs of actual dissections. Still others are composed of collections of artist renderings of organs or areas of interest. All rely on a basically two-dimensional (2-D) graphic display to depict and allow for a better understanding of a complicated 3-D structure.


Conus medullaris-cauda arteriovenous malformation and Klippel-Trenaunay syndrome: what is the treatment goal?
A 29-year-old man with Klippel-Trenaunay syndrome (KTS) presented with a symptomatic conus medullaris-cauda arteriovenous malformation (AVM) manifesting as back and right limb pain, which abruptly worsened with the onset of right limb weakness and urinary retention. He was treated by multisession endovascular embolization resulting in improved neurological status. KTS is a sporadic disease with unknown etiology, but genetic susceptibility may lead to the over-expression of angiogenic factors and increased angiogenesis. KTS may be exceptionally associated with slow-flow spinal AVM, but there is no consensus about the optimal treatment for these symptomatic lesions. Embolization treatment may represent a safe option to minimize complications and possibly improve the neurological status in patients with spinal AVM associated with KTS, if one or both legs are already impaired by hypertrophy or other vascular malformations. Genetic analysis may reveal an underlying angiogenesis change, so closer follow up might be indicated in selected patients.


Significance of platelet count in children admitted with bronchiolitis.
To determine the true prevalence of thrombocytosis in children less than 2 years of age with bronchiolitis, its association with risk factors, disease severity and thromboembolic complications. A retrospective observational medical chart review of 305 infants aged two years or less hospitalized for bronchiolitis. Clinical outcomes included disease severity, duration of hospital stay, admission to pediatric intensive care unit, or death. They also included complications of thrombocytosis, including thromboembolic complications such as cerebrovascular accident, acute coronary syndrome, deep venous thrombosis, pulmonary embolus, mesenteric thrombosis and arterial thrombosis and also hemorrhagic complications such as bleeding (spontaneous hemorrhage in the skin, mucous membranes, gastrointestinal, respiratory, or genitourinary tracts). The median age was 4.7 mo and 179 were males (59%). Respiratory syncytial virus was isolated in 268 (84%), adenovirus in 23 (7%) and influenza virus A or B in 13 (4%). Thrombocytosis (platelet count > 500 × 109/L) occurred in 88 (29%; 95%CI: 24%-34%), more commonly in younger infants with the platelet count declining with age. There was no significant association with the duration of illness, temperature on admission, white blood cell count, serum C-reactive protein concentration, length of hospital stay or admission to the intensive care unit. No death, thrombotic or hemorrhagic events occurred. Thrombocytosis is common in children under two years of age admitted with bronchiolitis. It is not associated with disease severity or thromboembolic complications.


Profitability of moderate intensive grazing of diary cows in the northeast.
Empirical analysis of the profitability of moderate grazing are presented based on sample dairy farms in Pennsylvania and New York. Net income per cow was higher for dairy farms that employed moderate intensive grazing than for dairy farms that employed extensive grazing. Income appeared to be adequate for family living expenses, but moderate intensive grazing could not be considered a high profit system. Detailed analyses of dairy farms that employed moderate and extensive grazing in northern Pennsylvania indicated that returns to management and owner equity were higher for pasture enterprises than for corn silage or hay enterprises. Positive dairy profits were related to lower feed costs. Milk production was lower on farms that employed moderate grazing than on farms that employed extensive grazing. Logit regression analysis characterized farms that employed moderate intensive grazing as oriented toward dairy rather than toward crop production; these farms had lower culling rates and a greater dependence on milk sales as a share of total sales. The reduced use of fertilizers and chemicals suggests that moderate grazing had environmentally sustainable features.


Effect of latanoprost on the diurnal variations in the intraocular and ocular perfusion pressure in normal tension glaucoma.
To evaluate the effects of latanoprost on the diurnal variations in the intraocular pressure (IOP) combined with the ocular perfusion pressure (OPP) in patients with normal tension glaucoma (NTG). Twenty-two eyes from 22 patients with NTG were used for the study. The diurnal variations in the IOP and blood pressure (BP) were measured every 3 hours without therapy, and then the patients were treated with latanoprost (0.005%) once daily for more than 12 weeks. The diurnal variations in the IOP and BP under medication were again measured every 6 hours. The diurnal variation of IOP for 24 hours, mean diurnal IOP, maximum IOP, minimum IOP, range of variation in IOP, OPP, and BP were compared between the baseline and after treatment by means of a paired t test. At 3 months after the start of the latanoprost treatment regimen, the IOP showed a statistically significant decrease at every assessed time point over 24 hours (P<0.001). Latanoprost significantly reduced the mean diurnal IOP, maximum IOP, minimum IOP, and mean range of variation in the IOP values from baseline (P<0.001, <0.001, <0.001, and 0.009, respectively). OPP after treatment showed no significant difference at any assessed time points from the baseline (P>0.1). Latanoprost did not significantly alter the mean diurnal OPP (P>0.1), and BP (P>0.5) from the baseline. Latanoprost was thus found to significantly reduce IOP over 24 hours, whereas it does not affect OPP and BP in NTG patients. Therefore, it may be a useful medication for NTG.


Important Roles of Water Clusters Confined in a Nanospace as Revealed by a Synchrotron X-ray Diffraction Study.
States of water molecules confined in a nanospace designed by montmorillonite (negatively charged silicate layer) and charge compensating benzylammonium were investigated. Caffeine was used as a probe because of its compatibility for the fine structure of the interlayer water. Powder synchrotron radiation X-ray diffraction (SXRD) and adsorption isotherms of the water vapor revealed a metastable structure of bimolecular water layers (2WLs) in the interlayer space. Water molecules readily penetrated to expand the interlayer space to 0.56 nm. The interlayer space did not increase further even in the presence of excess water. According to the isosteric heat of water, the expansion was limited because of moderate hydration as forming 2WLs. Caffeine molecules replaced a part of the water molecules in the 2WLs to expand the interlayer space to 0.65 nm. Time-resolved SXRD with an accumulation time of 500 ms revealed that the interlayer expansion reached a steady state within a few minutes. The caffeine intercalation proceeded, involving a change in the molecular orientation that increased the contact area of the caffeine molecules. The interlayer expansion was limited in all the solvents examined (mixtures of water with methanol, ethanol, acetone, and tetrahydrofuran), while the packing density of the incorporated caffeine was maximized in the absence of an organic solvent. The water molecules confined in the interlayer space acted as an actuator to accommodate a large quantity of amphiphilic molecules by adapting the nanostructure, which was achieved by releasing the confined water molecules.


Stereochemistry of 1,2-elimination and proton-transfer reactions: toward a unified understanding.
Many mechanistic and stereochemical studies have focused on the breaking of the C-H bond through base-catalyzed elimination reactions. When we began our research, however, chemists knew almost nothing about the stereospecificity of addition-elimination reactions involving conjugated acyclic carbonyl compounds, even though the carbonyl group is a pivotal functional group in organic chemistry. Over the last 25 years, we have studied the addition-elimination reactions of β-substituted acyclic esters, thioesters, and ketones in order to reach a comprehensive understanding of how electronic effects influence their stereochemistry. This Account brings together our understanding of the stereochemistry of 1,2-elimination and proton-transfer reactions, describing how each study has built upon previous work and contributed to our understanding of this field. When we began, chemists thought that anti stereospecificity in base-catalyzed 1,2-elimination reactions occurred via concerted E2 mechanisms, which provide a smooth path for anti elimination. Unexpectedly, we discovered that some E1cBirrev reactions produce the same anti stereospecificity as E2 reactions even though they proceed through diffusionally equilibrated, "free" enolate-anion intermediates. This result calls into question the conventional wisdom that anti stereochemistry must result from a concerted mechanism. While carrying out our research, we developed insights ranging from the role of historical contingency in the evolution of hydratase-dehydratase enzymes to the influence of buffers on the stereochemistry of H/D exchange in D2O. Negative hyperconjugation is the most important concept for understanding our results. This idea provides a unifying view for the largely anti stereochemistry in E1cBirrev elimination reactions and a basis for understanding the stereoelectronic influence of electron-withdrawing β-substituents on proton-transfer reactions.


Clinical characteristics of respiratory tract infection caused by Klebsiella pneumoniae in immunocompromised patients: a retrospective cohort study.
With advancements in medical technology and the growth of an aging society, the number of immunocompromised patients has increased progressively. Klebsiella pneumoniae (K. pneumoniae) is one of the most common opportunistic pathogens, causing a severe disease burden. We aimed to further clarify the differences in respiratory tract K. pneumoniae infections between immunocompromised and immunocompetent populations. We retrospectively compared cases of respiratory tract K. pneumoniae infection in immunocompromised and immunocompetent patients admitted to Ruijin Hospital in Shanghai between January 2019 and August 2020 to clarify the differences between the two groups. We enrolled 400 immunocompromised patients and 386 immunocompetent patients. Compared to the immunocompetent group, immunocompromised patients were more likely to develop bacteremia and shock and to require mechanical ventilation support during hospitalization. Immunocompromised patients also had a greater probability of polymicrobial infection and a higher rate of antibacterial resistance to carbapenem, which resulted in a higher intensive care unit admission rate, 30-day case fatality rate (CFR), and 6-month CFR. Multivariate analysis indicated that immunocompromised patients with respiratory diseases (odds ratio [OR], 2.189; 95% confidence interval [CI], 1.103-4.344; P = 0.025) and cardiovascular diseases (OR, 2.008; 95% CI, 1.055-3.822; P = 0.034), using mechanical ventilation (OR, 3.982; 95% CI, 2.053-7.722; P = 0.000), or infected with multidrug-resistant K. pneumoniae (OR, 3.870; 95%, 1.577-9.498; P = 0.003) were more likely to have a higher 30-day CFR. The disease burden of K. pneumoniae infection in immunocompromised patients is high. Immunocompromised patients who presented with respiratory diseases and cardiovascular diseases, used mechanical ventilation, or were infected with multidrug-resistant K. pneumoniae experienced a higher 30-day mortality rate.


Clinical significance of traumatic subarachnoid hemorrhage.
The clinical significance of traumatic subarachnoid hemorrhage (TSAH) was studied in 20 head-injured patients. They were classified into mild (9) and severe (11) groups by their initial Glasgow Coma Scale (GCS) scores. TSAH detected by computed tomography was localized in the Sylvian fissure in four of the nine mild group patients, but was also found in other basal subarachnoid cisterns in the other five. Except for one patient who developed delayed vasospasm and communicating hydrocephalus, all had favorable outcomes in this group. Massive TSAH was noted in the basal subarachnoid cisterns in 10 severe group patients and only one had a good outcome. Delayed intracerebral hematoma in the frontal or temporal lobe in the vicinity of the Sylvian fissure was found in two mild and two severe group patients with TSAH in the Sylvian fissures. Traumatic parenchymal lesions in the brainstem were minimal in one of the two autopsied patients and no evidence of diffuse axonal injury was found in both cases. Thus, TSAH in the Sylvian fissure is suggestive of focal brain contusion around the fissures. Massive TSAH in the basal subarachnoid cisterns is not necessarily associated with severe parenchymal injury of the brainstem.


Social support, temperament and previous prenatal loss interact to predict depression and anxiety during pregnancy.
Pregnancy is considered a complex period in a woman's life due to the changes that occur at different levels, which increase her vulnerability to developing psychological symptoms. A woman's temperament and perceived social support may play important roles in the development of such symptomatology. The main objective of the present work was to evaluate whether social support is an effective coping resource against the development of symptoms of depression and anxiety, especially among women with certain personality types, while also considering previous prenatal losses. The participants were 534 pregnant women in their 26th week of gestation. They completed measures related to social support, temperament (i.e. neuroticism and extraversion) and symptoms of depression and anxiety and informed us of any previous prenatal losses. The association between social support and depression was negative only for women who scored high on neuroticism. Extraversion did not interact with social support to predict depression or anxiety. Additionally, temperament interacted with previous prenatal losses, playing a relevant role in the development of anxiety symptoms. Previous losses had a greater effect on women who scored low in extraversion or high in neuroticism. Finally, a triple interaction between temperament, social support and previous prenatal loss emerged, indicating that previous prenatal loss was related to anxiety in women with low social support and low extraversion. Pregnant women, especially those who have suffered a previous prenatal loss and score high in neuroticism or low in extraversion, may benefit from interventions that enhance social support.


Persistent Chlamydia pneumoniae infection of cardiomyocytes is correlated with fatal myocardial infarction.
Acute myocardial infarction (AMI) associated with unfavorable prognosis is likely to be the consequence of a diffuse active chronic inflammatory process that destabilizes the whole coronary tree and myocardium, suggesting a possible common causal agent underlying both conditions. The main objective of this study was to investigate whether Chlamydia pneumoniae (CP) infection occurred beyond the coronary plaques, namely in the myocardium of individuals who died of AMI. The presence of CP cell wall antigen (OMP-2) and CP-HSP60 was investigated in the myocardium and coronary plaques of 10 AMI and 10 age-matched control patients by immunohistochemistry, electron microscopy, and molecular biology. OMP-2 antigens were found in the unaffected myocardium of 9 of 10 AMI patients. Conversely, only 1 of 10 control patients exhibited a positive staining for CP. Moreover, OMP-2 and CP-HSP60 were detected in the whole coronary tree. CP presence was strongly associated with a T-cell inflammatory infiltrate. Our results suggest that CP may underlie both coronary and myocardial vulnerabilities in patients who died of AMI and corroborate the notion that CP may act by reducing cardiac reserves, thus worsening the ischemic burden of myocardium.


Caregivers' Expectations on Possible Functional Changes following Disease-Modifying Treatment in Type II and III Spinal Muscular Atrophy: A Comparative Study.
Background: The primary aim of this study was to explore current caregivers' expectations on possible functional changes following treatment in comparison to data obtained in the pre-pharmacological era. Methods: A questionnaire, previously used in 2016, was administered to caregivers of type II and III SMA patients of age between 3 and 71 years, and to patients over the age of 13 years. The questionnaire focuses on (1) caregivers and patients expectations, (2) meaningfulness of the changes observed on the functional motor scales, and (3) their willingness to be enrolled in a clinical trial. A comparative study was performed with data obtained using the same questionnaire soon before the advent of disease-modifying therapies. Results: We administered the questionnaire to 150 caregivers. When comparing current caregiver data to those obtained in 2016, the most obvious differences were related to disease perception over the last year (stability: 16.5% in 2016 vs. 43.6% in 2022; deterioration 70.5% vs. 12.8%, and improvement: 12.9% vs. 43.6%) and expectations from clinical trials with higher expectations in 2022 compared to 2016 (p < 0.001). Forty-five of the 150 in the current study were caregivers of patients above the age of 13. In these 45 the questionnaire was also administered to the patient. No difference was found in responses between patients and their caregivers. Conclusions: Both carers and patients reported that even small changes on functional scales, similar to those reported by clinical studies and real-world data, are perceived as meaningful. Comparing the recent responses to those obtained in 2016, before pharmacological treatment was available, we found significant changes in caregivers' perception with increased expectations. These findings will provide a better understanding of the patients' expectations and facilitate discussion with regulators.


Repair of Physiologic Time Series: Replacement of Anomalous Data Points to Preserve Fractal Exponents.
Extraction of fractal exponents via the slope of the power spectrum is common in the analysis of many physiological time series. The fractal structure thus characterized is a manifestation of long-term correlations, for which the temporal order of the sample values is crucial. However, missing data points due to artifacts and dropouts are common in such data sets, which can seriously disrupt the computation of fractal parameters. We evaluated a number of methods for replacing missing data in time series to enable reliable extraction of the fractal exponent and make recommendations as to the preferred replacement method depending on the proportion of missing values and any a priori estimate of the fractal exponent.


Moderate exercise in mice improves cancer plus chemotherapy-induced muscle wasting and mitochondrial alterations.
Cancer cachexia is a multifactorial syndrome characterized by anorexia, body wasting, and muscle and adipose tissue loss, impairing patient's tolerance to anticancer treatments and survival. The aim of the present study was to compare the effects induced in mice by tumor growth alone (C26) or in combination with chemotherapy [C26 oxaliplatin and 5-fluorouracil (oxfu)] and to evaluate the potential of moderate exercise. Oxfu administration to C26 mice exacerbated muscle wasting and triggered autophagy or mitophagy, decreased protein synthesis, and induced mitochondrial alterations. Exercise in C26 oxfu mice counteracted the loss of muscle mass and strength, partially rescuing autophagy and mitochondrial function. Nevertheless, exercise worsened survival in C26 oxfu mice in late stages of cachexia. In summary, chemotherapy further impinges on cancer-induced alterations, worsening muscle wasting. An ideal multifactorial and early intervention to prevent cancer cachexia could take advantage of exercise, improving patient's energy metabolism, mobility, and quality of life.-Ballarò, R., Beltrà, M., De Lucia, S., Pin, F., Ranjbar, K., Hulmi, J. J., Costelli, P., Penna, F. Moderate exercise in mice improves cancer plus chemotherapy-induced muscle wasting and mitochondrial alterations.


Micro X-ray diffraction analysis of thin films using grazing-exit conditions.
An X-ray diffraction technique using a hard X-ray microbeam for thin-film analysis has been developed. To optimize the spatial resolution and the surface sensitivity, the X-ray microbeam strikes the sample surface at a large glancing angle while the diffracted X-ray signal is detected with a small (grazing) exit angle. Kirkpatrick-Baez optics developed at the Photon Factory were used, in combination with a multilayer monochromator, for focusing X-rays. The focused beam size was about 10 x 10 micro m. X-ray diffraction patterns of Pd, Pt and their layered structure were measured. Using a small exit angle, the signal-to-background ratio was improved due to a shallow escape depth. Under the grazing-exit condition, the refraction effect of diffracted X-rays was observed, indicating the possibility of surface sensitivity.


Development and Content Validity of the Bilateral Vestibulopathy Questionnaire.
To date, the burden and severity of the full spectrum of bilateral vestibulopathy (BVP) symptoms has not yet been measured in a standardized manner. Since therapeutic interventions aiming to improve BVP symptoms are emerging, the need for a new standardized assessment tool that encompasses the specific aspects of BVP arises. Therefore, the aim of this study was to develop a multi-item Patient Reported Outcome Measure (PROM) that captures the clinically important symptoms of BVP and assesses its impact on daily life. The development of the Bilateral Vestibulopathy Questionnaire (BVQ) consisted of two phases: (I) initial item generation and (II) face and content validity testing. Items were derived from a literature review and individual semi-structured interviews focusing on the full spectrum of reported BVP symptoms (I). Subsequently (IIa), individual patient interviews were conducted using "thinking aloud" and concurrent verbal probing techniques to assess the comprehensibility of the instructions, questions and response options, and the relevance, missing domains, or missing items. Interviews continued until saturation of input was reached. Finally, international experts with experience in the field of the physical, emotional, and cognitive symptoms of BVP participated in an online focus group to assess the relevance and comprehensiveness of the BVQ (IIb). The BVQ consisted of two sections. The first section included 50 items scored on a six-point Likert scale arranged into seven constructs (i.e., imbalance, oscillopsia, other physical symptoms, cognitive symptoms, emotional symptoms, limitations and behavioral changes and social life). The second section consisted of four items, scored on a visual analog scale from 0 to 100, to inquire about limitations in daily life, perceived health and expectations regarding future recovery. Interviews with BVP patients [n = 8, 50% female, mean age 56 years (range 24-88 years)] and the expert meeting confirmed face and content validity of the developed BVQ. The BVQ, which was developed to assess the spectrum of BVP symptoms and its impact on daily life, proved to have good face and content validity. It can be used to characterize current self-reported symptoms and disability and to evaluate symptom burden before and after therapeutic interventions in future research and clinical practice.


Acylhomoserine lactone production and degradation by the fish pathogen Tenacibaculum maritimum, a member of the Cytophaga-Flavobacterium-Bacteroides (CFB) group.
Tenacibaculum maritimum (formerly Flexibacter maritimus) is a filamentous, biofilm-forming member of the Cytophaga-Flavobacterium-Bacteroides group (or Bacteroidetes), which causes the widely distributed marine fish disease tenacibaculosis. A search for N-acylhomoserine lactones (AHLs) quorum-sensing (QS) signals in the culture media of nine representative strains of this species using different biosensor strains revealed the presence of short-type AHL activity in all of them. N-butyryl-L-homoserine lactone (C4-HSL) was identified in T. maritimum NCIMB2154(T) by LC-MS. A degradation activity for long-acyl AHLs (C10-HSL) was subsequently demonstrated in T. maritimum NCIMB2154(T). The acidification of the culture medium after degradation did not allow the recovery of C10-HSL, which indicates a possible acylase-type degradation activity. Even though the physiological processes under the control of AHL-mediated QS in T. maritimum need to be further characterized, this discovery extends the paradigm of AHL-mediated QS signalling beyond the Proteobacteria and reinforces its ecological significance.


Knowledge-based reconstruction for measurement of right ventricular volumes on cardiovascular magnetic resonance images in a mixed population.
Follow-up of right ventricular performance is important for patients with congenital heart disease. Cardiac magnetic resonance imaging is optimal for this purpose. However, observer-dependency of manual analysis of right ventricular volumes limit its use. Knowledge-based reconstruction is a new semiautomatic analysis tool that uses a database including knowledge of right ventricular shape in various congenital heart diseases. We evaluated whether knowledge-based reconstruction is a good alternative for conventional analysis. To assess the inter- and intra-observer variability and agreement of knowledge-based versus conventional analysis of magnetic resonance right ventricular volumes, analysis was done by two observers in a mixed group of 22 patients with congenital heart disease affecting right ventricular loading conditions (dextro-transposition of the great arteries and right ventricle to pulmonary artery conduit) and a group of 17 healthy children. We used Bland-Altman analysis and coefficient of variation. Comparison between the conventional method and the knowledge-based method showed a systematically higher volume for the latter group. We found an overestimation for end-diastolic volume (bias -40 ± 24 mL, r = .956), end-systolic volume (bias -34 ± 24 mL, r = .943), stroke volume (bias -6 ± 17 mL, r = .735) and an underestimation of ejection fraction (bias 7 ± 7%, r = .671) by knowledge-based reconstruction. The intra-observer variability of knowledge-based reconstruction varied with a coefficient of variation of 9% for end-diastolic volume and 22% for stroke volume. The same trend was noted for inter-observer variability. A systematic difference (overestimation) was noted for right ventricular size as assessed with knowledge-based reconstruction compared with conventional methods for analysis. Observer variability for the new method was comparable to what has been reported for the right ventricle in children and congenital heart disease with conventional analysis.


Suprabrow minicraniotomy for suprasellar tumours.
We present our experience with suprabow minicraniotomy in the excision of 18 suprasellar tumours between May 2001 and September 2003. There were 14 females and four males with ages ranging from 7 to 59 years. The tumours included one epidermoid cyst, 10 meningiomas, five craniopharyngiomas and two optico-chiasmatico-hypothalamic (OCHG) gliomas. The size of the one-piece craniotomy was about 2.5 cm. Both OCHGs were biopsied; one craniopharyngioma was totally excised, the remaining craniopharyngiomas and epidermoid cyst were subtotally excised. Four meningiomas were radically excised and six were subtotally removed. The patient with a craniopharyngioma that had a radical excision died of hypothalamic dysfunction. One patient developed a postoperative meningitis and subsequent hydrocephalus requiring a shunt. The other patients did well and the cosmetic result was excellent in all cases. This basal approach through a small craniotomy provided good surgical access to suprasellar tumours with minimal brain retraction.


The Diagnosis of Intrapulmonary Metastasis Multifocal Pulmonary Ground-Glass Nodules Based on Oncogenic Driver Mutation: Two Case Reports and Review of Literature.
With the increased use of low-dose computed tomography (LDCT), the detection of multifocal pulmonary ground-glass nodules (GGNs) has increased. According to the current clinical guidelines, multifocal GGNs tend to be treated as the multiple primary early-stage lung adenocarcinoma. However, studies have indicated that parts of multiple GGNs may originate from single nodules via intrapulmonary metastasis (IPM). Such IPM indicates the advanced stages even when the multiple GGNs are present as the early characteristics in pathological assessments. However, no gold standard exists for the differential diagnosis of multiple IPM GGNs. Here, we report two multifocal pulmonary GGNs cases where panel sequencing (672 driver mutation loci) showed that patient 1 (P1) shared two rare epidermal growth factor receptor (EGFR) mutations (primary L747_T751del and primary T790M) in the left upper lobe anterior segment and left lower lobe superior segment, respectively. Patient 2 (P2) shared a low-frequency human epidermal growth factor receptor 2 (HER2) mutation (primary Tyr772_Ala775dup) in two GGNs located in the apicoposterior and superior lingular segment of the left lower lobe (LLL). Oncogenic driver mutations were concordant between primary tumors and metastasis. Thus, shared low-frequency driver mutations in multiple GGNs strongly suggested that IPM existed with a high probability in these patients. Also, tumor cell spread through air spaces (STAS) was identified in pathological sections of the left upper lobe (LUL) nodule of P1, suggesting aerogenous spread may have been an effective pathway for IPM. Our report suggests that oncogenic driver mutations have prominent diagnostic value for IPM. Also, GGN IPM may occur in one lung lobe and between in different lung lobes.


Tuberculous pericardial effusion: features, tamponade, and computed tomography.
The clinical features with particular reference to tamponade and mediastinal adenopathy were studied in tuberculous pericardial effusion. Tamponade is a frequent complication and the recognition of tuberculous etiology can be difficult. Involvement of the pericardium is mostly from mediastinal lymph nodes that have not been studied. This was a prospective cohort study. All patients had large effusions, and underwent pericardiocentesis and chest computed tomography. Patients with tuberculosis had specific therapy. Others with viral/idiopathic effusion served as controls for the computed tomography studies. There were 26 patients with tuberculosis: 18 had tamponade on echocardiography. All had symptoms. Fever (n = 23) and dyspnea (n = 20) were the most frequent presenting symptoms. Pericardial rub was heard in 14, and 3 had enlarged cervical or axillary nodes. Pulmonary tuberculosis was present in 6. Tuberculin skin test measured 17 +/- 3.3 mm. The biopsy specimen showed a granuloma in 22 of 24. All 26 had mediastinal lymph nodes > 10 mm with a mean size of 19.5 +/- 8.6 mm that disappeared (81%) or regressed (19%) on treatment (p < 0.001). Aortopulmonary nodes were most frequently enlarged (65.4%) and hilar the least. Three required pericardiectomy. At follow-up all were doing well. None with viral/idiopathic effusion had lymph node enlargement. Fever, dyspnea, and tamponade were frequent with tuberculosis. The prognosis was good with specific therapy. Mediastinal nodes were enlarged in all and only with tuberculosis and not with viral/idiopathic effusion. Nodes disappeared or regressed with treatment. In the appropriate clinical context, mediastinal lymph node enlargement on chest computed tomography along with a strongly positive skin test results could help in the diagnosis of a tuberculous etiology of pericardial effusion.


Cloning and characterization of the deoxyribonuclease sd alpha gene from Streptococcus pyogenes.
The pathogenesis of Streptococcus pyogenes infection, especially toxic shock-like syndrome (TSLS), is still not fully understood; however, the exoproteins have been considered to play a role. We analyzed the culture supernatant proteins (exoproteins) from a TSLS-related isolate belonging to M3 serotype S. pyogenes by two-dimensional gel electrophoresis and characterized a single protein spot by using BLAST database. We cloned the gene of this protein and named it sd alpha, which was similar to the deoxyribonuclease (DNase) sdc of S. equisimilis. We showed that the recombinant protein from the sd alpha gene had DNase activity. By polymerase chain reaction, we found that the sd alpha gene was present in most clinically isolated S. pyogenes including TSLS-related isolates. We thus conclude that Sd alpha is a new DNase of S. pyogenes.


Remifentanil for sedation and analgesia in a preterm neonate with respiratory distress syndrome.
We present the efficacy and safety of the use of remifentanil for intubation, sedation and analgesia in a preterm infant during mechanical ventilation for respiratory distress syndrome. A 34-week-old baby, born by cesarean delivery that developed respiratory distress, required intubation and ventilatory support. For intubation, the baby was given midazolam (0.2 mg.kg(-1)) and remifentanil (1 microg.kg(-1)). The intubation conditions were assessed and classified as excellent. The remifentanil infusion was started at dose 0.75 microg.kg(-1).min(-1) and the dose adjustments were made depending on the neonatal infant pain scale (NIPS), hemodynamic and respiratory changes or the presence of spontaneous movements. Pulse oximetry, respiratory rate, ECG and invasive blood pressure were continuously monitored. He was given surfactant within 2.5 h of life after which ventilator parameters could be progressively decreased. Three hours later, the remifentanil infusion was decreased to 0.5 microg.kg(-1).min(-1), and he remained sedated (NIPS < 2). Six hour after surfactant administration, blood gases and chest X ray were normal. The remifentanil infusion was then discontinued and 30 min later the baby was awake and extubated with success. There were no side effects after intubation or during the continuous infusion. The profile of remifentanil allowing a rapid recovery, the absence of side effects and a good level of sedation and analgesia support the choice of this opioid for sedation in the NICU.


Cysteine as a Monothiol Reducing Agent to Prevent Copper-Mediated Oxidation of Interferon Beta During PEGylation by CuAAC.
Bioconjugation by copper-catalyzed azide-alkyne cycloaddition (CuAAC) provides a powerful means to produce site-specifically modified proteins. However, the use of a copper catalyst brings about the possible generation of reactive oxygen species that could cause degradation of vulnerable amino acid residues. We investigated whether PEGylation by CuAAC caused any modifications to the therapeutic protein interferon beta-1b, which was produced via global amino acid substitution with azidohomo-alanine at the N-terminus and contains no methionine residues. Using previously reported reaction conditions, LC-MS peptide mapping detected +32 Da and +48 Da oxidation modifications of tryptic peptides 28-33 (LEYCLK) and 137-147 (EYSHCAWTIVR) in the protein post-PEGylation. The oxidative degradation increased with reaction time, whereas reducing the copper concentration slowed the PEGylation rate as well as the oxidation rate. Replacing dithiothreitol (DTT) with any of five different monothiol reducing agents in anaerobic conditions allowed efficient PEGylation in 2-4 h and abrogated oxidative degradation. Free cysteine provided reproducible reaction results as a reducing agent in this system and has been successfully applied to other protein conjugations. Monothiol reducing agents, such as cysteine, may be useful tools as protective reducing agents for CuAAC in some bioconjugation systems.


Inhibition of vascular endothelial growth factor reduces cardiac allograft vasculopathy.
Cardiac allograft vasculopathy (CAV) is a leading expression of chronic organ rejection at and beyond 1 year post-transplantation. Host bone marrow (BM)-derived cell migration to the allograft has been demonstrated in earlier work. Vascular endothelial growth factor (VEGF) is endogenously overexpressed within allografts. Graft neo-angiogenesis has been proposed as a mechanism by which VEGF may contribute to CAV. Herein we assess the therapeutic effect of inhibition of VEGF expression in CAV. In 129J mice, female donor hearts were heterotopically transplanted into C57/B16 males and treated with soluble VEGF receptor 1 (sVEGFR1) or vehicle control. The effect of VEGF inhibition on BM-mediated microvascular outgrowth and endothelial cell migration and proliferation were assessed using in vitro assays of aortic ring angiogenesis, wound healing and proliferation, respectively. At 21 days post-transplantation, treatment with sVEGFR1 significantly reduced both percent luminal narrowing (p < 0.05) and percent of vessels affected (p < 0.005). sVEGFR1 significantly reduced average wet heart weight (p < 0.05), whereas mean ventricular cross-sectional area remained similar. Treatment of aortic rings with both sVEGFR1 and VEGFR2 tyrosine phosphorylation inhibitor (Ki 8751) significantly reduced BM-mediated microvascular outgrowth length (p < 0.05) and area (p < 0.05). Treatment of human coronary artery endothelial cells with sVEGFR1 and Ki 8751 significantly reduced BM-mediated endothelial cell migration (p < 0.005) and proliferation (p < 0.05). VEGF inhibition reduces the severity and incidence of CAV in mouse models of cardiac transplantation, while attenuating myocardial edema and neo-angiogenesis. Using this model, we provide in vitro evidence of the role of VEGF signaling in BM-mediated microvascular outgrowth and endothelial cell migration and proliferation. VEGF inhibition may represent a novel approach to CAV treatment and prevention.


Pleasantness of facial profile and its correlation with soft tissue cephalometric parameters: Perception of orthodontists and lay people.
This study was aimed to evaluate the perception of orthodontists and of lay people about the facial profile and its possible correlation with cephalometrics parameters. A total of 20 evaluators were divided into two groups (10 orthodontists and 10 people with no relation to such area - lay people). They were asked to evaluate the photographs of 25 young males and of 25 young females, aged 17-24-year-old (mean age of 22.3 years, standard deviation 2.41 years). Photographs were randomly arranged in a photo album. The evaluators rated each photograph by means of a scale ranging from "good" to "deficient" based on the pleasantness of their facial profile. Nasolabial angle, Holdaway's H-line and the distance from H-line to nose tip were measured, in order to verify a possible relation between these soft tissue profile cephalometric measurements and the subjective ratings. The kappa statistics test showed a concordance of 0.23 among orthodontists and 0.24 among lay people. Regarding the perception of orthodontists and lay people on facial profile, no significant divergence could be detected. For the correlation between cephalometric parameters and subjective ratings, there was a statistically significant correlation between the measures H and H-nose and the rating ascribed to the profile. It was concluded that smaller the difference from the normal cephalometric pattern, the higher was the rating given to the profile, demonstrating an important relation between subjective and objective criteria.


Present Status of the UVSOR Facility - 1997.
The UVSOR has been operational for nearly 14 years. In recent years, improvements to accelerators and beamlines have been carried out to obtain better performance and new scientific achievements. For example, a new type of helical undulator and a resonance transverse kicker were installed into the straight sections of the storage ring, and about one-third of the beamlines were upgraded by installing new monochromators and an interferometer. In addition, instruments and equipment were improved for research studies, with many interesting results. The most up-to-date information of the UVSOR facility is presented.


"Every path will end in darkness" or: why psychoanalysis needs metapsychology.
This article focuses on the dialectic of metapsychology and hermeneutics in psychoanalysis. By combining the causal language of the former with the intentional terminology of the latter, Freud's discourse continuously transgresses the narrowly conceived boundaries of scientific disciplines and places its stakes in both the humanities and the natural sciences. The argument is made that attempts to reduce psychoanalytic theory to either causal explanation or interpretation of meaning turn it into a closed thought system and rob it of its vitality. It is argued, moreover, that although Freud understood himself to be a scientist, by eschewing the dichotomous reductionism characteristic of both his orthodox followers and critics who tried and still try to turn psychoanalysis into either a natural-science-like discipline or a hermeneutics, Freud demonstrated that his self-understanding was far more sophisticated than admitted by either of these two groups. This argument is supported by a detailed discussion of Freud's epistemological premises, his conception of science and reality, and especially, the place he allocated to metapsychology in his interdisciplinary science. It is claimed that metapsychology served Freud as a double-edged sword, both enabling creative and metaphorical thought about the mind's hidden reality and revealing the necessary incompleteness of hermeneutics. The article concludes with the claim that psychoanalysis needs metapsychology in order to pursue this dual task.


Do weekend oocyte retrievals have an impact on the intracytoplasmic sperm injection cycles outcome?
Despite the fact that ovarian stimulation is controlled, it is not always predictable. Because the day of human chorionic gonadotrophin (hCG) injection depends mainly on the patient's ovarian response to gonadotrophins, the day of oocyte retrieval cannot be determined in advance. As a result, oocyte retrievals are often scheduled to occur on weekends, a fact that entails at least one extra working day for the staff, and could lead to physical and psychological stress, especially in embryologists. The aim of this study was to evaluate whether intracytoplasmic sperm injection (ICSI) outcomes are influenced by the day of oocyte retrieval. A total of 327 ICSI cycles, whose retrievals were performed on Wednesdays and Sundays, were analysed in this retrospective study. Cycles were subdivided into two groups according to the day of oocyte retrieval: group W (n = 196), cycles in which oocyte retrieval was performed on Wednesday; and group S (n = 131), cycles in which oocyte retrieval was performed on Sunday. Groups were compared regarding fertilization, implantation, pregnancy and take-home baby rates. No significant differences were observed between groups A and B regarding fertilization rate (68.9% and 72.5%; p = 0.1589), implantation rate (21.8% and 24.3%; p = 0.5714), pregnancy rate (29.9% and 31.6%; p = 0.7129) and take-home baby rate (23.6% and 28.1%; p = 0.4351). A well-trained embryologist's group adhering to staff scheduling allows large programmes to ensure a similar outcome independent of the workload or workday on which embryologists perform the manipulation of gametes.


Saccharomyces cerevisiae RAD2 gene: isolation, subcloning, and partial characterization.
A plasmid (pNF2000) containing a 9.7-kilobase pair DNA insert that complements the UV sensitivity of rad2-1, rad2-2, and rad2-4 mutants of Saccharomyces cerevisiae has been isolated from a yeast genomic library. Genetic analysis of strains derived by transformation of rad2 mutants with an integrating plasmid containing a 9.3-kilobase pair fragment from pNF2000 shows that the fragment integrates exclusively at the chromosomal rad2 gene. We therefore conclude that this plasmid contains the RAD2 gene. The 9.3-kilobase pair fragment was partially digested with Sau3A and cloned into a multicopy yeast vector designed for easy retrieval of Sau3A inserts. The smallest subclone that retains the RAD2 gene is 4.5 kilobase pairs. This fragment was partially digested with Sau3A and cloned into an integrating plasmid. These plasmids were isolated and integrated into a heterozygous rad2/RAD2 strain. Plasmids containing internal fragments of the RAD2 gene were identified because they yielded UV-sensitive transformants due to disruption of the RAD2 gene. Sporulation of diploids transformed with integrating plasmids containing internal fragments of RAD2 gave rise to four viable haploids per tetrad, indicating that unlike the RAD3 gene of S. cerevisiae, the RAD2 gene is not essential for the viability of haploid cells under normal growth conditions. Measurements of the RNA transcript by RNA-DNA hybridization with the internal fragment as the probe indicate a size of approximately 3.2 kilobases.


C677T (thermolabile alanine/valine) polymorphism in methylenetetrahydrofolate reductase (MTHFR): its frequency and impact on plasma homocysteine concentration in different European populations. EARS group.
A common polymorphism has been described in the methylenetetrahydrofolate reductase (MTHFR) gene, substituting an alanine (A) for a valine (V), where the V allele results in a thermolabile enzyme with reduced activity. This polymorphism is easily detectable by PCR amplification and digestion with HinfI restriction enzyme. We describe the use of the MADGE high throughput genotyping system for rapid typing of this polymorphism. Seven hundred and eighty five individuals participating in the European Atherosclerosis Research Study II (EARS II), aged 22-25 from 14 universities in 12 countries across Europe were genotyped for this polymorphism. The frequency of the V allele was 0.32 overall (95% CI; 0.30-0.35), but was significantly lower in the Baltic countries (0.23; 95% CI; 0.19-0.28) compared with the other regions of Europe (0.37; 95% CI; 0.32-0.38) (P < 0.001). Individuals homozygous for the V allele had statistically significant (P < 0.001) higher plasma homocysteine (16.5 micromol/l) compared with those heterozygous for an A allele (10.4 micromol/l) or homozygous for an A allele (10.0 micromol/l). This effect was seen in all countries and regions of Europe. Mean plasma homocysteine levels were significantly higher in the South compared to the Baltic, UK and Middle regions (P = 0.001), but this difference was not explained by the difference in the frequency of the V allele in the samples. This polymorphism explained 12.3% of the total sample variance in plasma homocysteine, other measured factors (smoking, alcohol consumption, systolic blood pressure, physical activity) explained 0.7%. This study demonstrates the large and consistent impact of the thermolabile MTHFR variant on plasma homocysteine levels in different European populations, and shows a regional difference in the levels of homocysteine that must be explained by other genetic or environmental factors.


Radiography in acute ankle injuries: the Ottawa Ankle Rules versus local diagnostic decision rules.
We validate the Ottawa Ankle Rules and 2 Dutch ankle rules in distinguishing clinically significant fractures from insignificant fractures and other injuries in patients with a painful ankle presenting to the emergency department. This prospective comparison of 3 ankle rules was conducted in the ED of a 580-bed community teaching hospital in Amsterdam from January 1998 to April 1999. Participants included 647 consecutive patients aged 18 years or older presenting with a painful ankle after trauma. All physicians received extensive and pictorial training on how to correctly score the respective items of the rules. The physician on call recorded these items derived from history and physical examination on a standardized data sheet. All patients subsequently underwent standard radiographic assessment. A radiologist and a trauma surgeon evaluated the radiographs blinded from the results of the data sheet form and the treatment given. The diagnostic performance of the 3 rules was measured in terms of sensitivity, specificity, and the reduction of radiographs. Receiver operating characteristic (ROC) curves were constructed, and the area under the ROC curves was calculated and compared. Seventy-four fractures were seen, of which 41 were clinically significant. The Ottawa Ankle Rules had a sensitivity of 98% for identifying clinically significant fractures; the local rules scored 88% and 59%, respectively. The potential savings in radiographs for the 3 decision rules were 24%, 54%, and 82%, respectively. The area under the ROC curve was better for both the local rules (0.84 and 0.83) compared with the Ottawa Ankle Rules (0.76). Because the identification of all relevant fractures is more important than a reduction in radiographs, the higher sensitivity of the Ottawa Ankle Rules makes these most suitable for implementation in The Netherlands.


Adverse outcomes of poor micronutrient status in childhood and adolescence.
The adverse effects of micronutrient deficiencies and excesses in children up to reproductive age are presented. A summary of risks and adverse functional and health outcomes associated with deficient and excessive intakes and nutrition status of iron, iodine, zinc, vitamins A and D, folate, vitamin B12, and riboflavin is presented. Nutrient-nutrient interactions of micronutrients, age, gender, and other host and environmental conditions, such as pregnancy, genetic conditions, overall nutrition, force of infection, and social conditions are considered as covariates in trying to define causation and outcomes due to specific micronutrients. The outcomes analyzed focus on growth and development, mental and neuromotor performance, immunocompetence, physical working capacity, morbidity, and in the case of pregnancy, overall reproductive performance. The results presented include responses to specific and multiple "experimental" interventions. A brief analysis of possible public health programs is presented, with emphasis on prevention.


Development of Engineered Bacteriophages for Escherichia coli Detection and High-Throughput Antibiotic Resistance Determination.
T7 bacteriophages (phages) have been genetically engineered to carry the lacZ operon, enabling the overexpression of beta-galactosidase (β-gal) during phage infection and allowing for the enhanced colorimetric detection of Escherichia coli (E. coli). Following the phage infection of E. coli, the enzymatic activity of the released β-gal was monitored using a colorimetric substrate. Compared with a control T7 phage, our T7lacZ phage generated significantly higher levels of β-gal expression following phage infection, enabling a lower limit of detection for E. coli cells. Using this engineered T7lacZ phage, we were able to detect E. coli cells at 10 CFU·mL-1 within 7 h. Furthermore, we demonstrated the potential for phage-based sensing of bacteria antibiotic resistance profiling using our T7lacZ phage, and subsequent β-gal expression to detect antibiotic resistant profile of E. coli strains.


A2BAR Antagonism Decreases the Glomerular Expression and Secretion of Chemoattractants for Monocytes and the Pro-Fibrotic M2 Macrophages Polarization during Diabetic Nephropathy.
Some chemoattractants and leukocytes such as M1 and M2 macrophages are known to be involved in the development of glomerulosclerosis during diabetic nephropathy (DN). In the course of diabetes, an altered and defective cellular metabolism leads to the increase in adenosine levels, and thus to changes in the polarity (M1/M2) of macrophages. MRS1754, a selective antagonist of the A2B adenosine receptor (A2BAR), attenuated glomerulosclerosis and decreased macrophage-myofibroblast transition in DN rats. Therefore, we aimed to investigate the effect of MRS1754 on the glomerular expression/secretion of chemoattractants, the intraglomerular infiltration of leukocytes, and macrophage polarity in DN rats. Kidneys/glomeruli of non-diabetic, DN, and MRS1754-treated DN rats were processed for transcriptomic analysis, immunohistopathology, ELISA, and in vitro macrophage migration assays. The transcriptomic analysis identified an upregulation of transcripts and pathways related to the immune system in the glomeruli of DN rats, which was attenuated using MRS1754. The antagonism of the A2BAR decreased glomerular expression/secretion of chemoattractants (CCL2, CCL3, CCL6, and CCL21), the infiltration of macrophages, and their polarization to M2 in DN rats. The in vitro macrophages migration induced by conditioned-medium of DN glomeruli was significantly decreased using neutralizing antibodies against CCL2, CCL3, and CCL21. We concluded that the pharmacological blockade of the A2BAR decreases the transcriptional expression of genes/pathways related to the immune response, protein expression/secretion of chemoattractants, as well as the infiltration of macrophages and their polarization toward the M2 phenotype in the glomeruli of DN rats, suggesting a new mechanism implicated in the antifibrotic effect of MRS1754.