Stabilization of 2D graphene, functionalized graphene, and Ti2CO2 (MXene) in super-critical CO2: a molecular dynamics study.
The stabilization of nanoparticles is a main concern to produce an efficient nanofluid. Here, we carry out Molecular Dynamics (MD) modeling to evaluate the stabilization of 2D graphene, oxygen- and hydrogen-functionalized graphene, and Ti2CO2 MXene nano-sheets in carbon dioxide (CO2) liquid under supercritical conditions. The rheological properties (i.e., viscosity and self-diffusion) of the nanofluids with the corresponding nano-sheets are also calculated. The results show that pristine graphene aggregates in the liquid and the adsorbed CO2 molecules on the graphene surfaces could not provide enough repulsive forces to avoid their aggregation. The stabilization of graphene is improved by surface functionalization of graphene with oxygen and hydrogen functional groups. This is because, first, the presence of the corresponding surface groups can alter the electrostatic forces of the graphene nano-sheets and, second, a larger number of CO2 molecules attracted by the functionalized nano-sheets can provide additional repulsive forces between the nano-sheets. The results demonstrate that Ti2CO2 MXene nano-sheets aggregate severely although they are covered by oxygen surface terminations and attract more CO2 molecules than the pristine graphene. This is attributed to the strong interlayer coupling between the MXene nano-sheets so that the electrostatic repulsion of the MXene surfaces could not overcome the interlayer coupling. Based on our results, the stabilization of the corresponding 2D nano-sheets in CO2 liquid is in the order: oxygen-functionalized graphene > hydrogen-functionalized graphene > pristine graphene > Ti2CO2 MXene. The calculation of the rheological properties of the nanofluids with different nano-sheets demonstrates that immersing the nano-sheets in the CO2 liquid affords a superior viscosity enhancement as much as possible to be dispersed in the liquid. This study expands the potential applications of these 2D materials in producing SC-CO2 based nanofluids.


Atorvastatin Promotes Phagocytosis and Attenuates Pro-Inflammatory Response in Human Retinal Pigment Epithelial Cells.
Phagocytosis of daily shed photoreceptor outer segments is an important function of the retinal pigment epithelium (RPE) and it is essential for retinal homeostasis. RPE dysfunction, especially impairment of its phagocytic ability, plays an essential role in the pathogenesis of age-related macular degeneration (AMD). Statins, or HMG CoA (3-hydroxy-3-methylglutaryl-coenzyme A) reductase inhibitors, are drugs with multiple properties that have been extensively used to treat hyperlipidemia. However, their effect on RPE cells has not been fully elucidated. Here we report that high dose atorvastatin increased the phagocytic function of ARPE-19 cells, as well as rescue the cells from the phagocytic dysfunction induced by cholesterol crystals and oxidized low-density lipoproteins (ox-LDL), potentially by increasing the cellular membrane fluidity. Similar effects were observed when evaluating two other hydrophobic statins, lovastatin and simvastatin. Furthermore, atorvastatin was able to block the induction of interleukins IL-6 and IL-8 triggered by pathologic stimuli relevant to AMD, such as cholesterol crystals and ox-LDL. Our study shows that statins, a well-tolerated class of drugs with rare serious adverse effects, help preserve the phagocytic function of the RPE while also exhibiting anti-inflammatory properties. Both characteristics make statins a potential effective medication for the prevention and treatment of AMD.


Systematic review and directors survey of quality indicators for the cardiovascular intensive care unit.
Quality indicators (QIs) are increasingly used in cardiovascular care as measures of performance but there is currently no consensus on indicators for the cardiovascular intensive care unit (CICU). We searched Medline, CINAHL, EMBASE, and COCHRANE databases from inception until October 2016 and websites for organizations involved in quality measurement for QIs relevant to cardiovascular disease in an intensive or critical care setting. We surveyed 14 expert cardiac intensivist-administrators (7 European; 7 North American) on the importance and relevance of each indicator as a measure of CICU care quality using a scale of 1 (=lowest) to 10 (=highest). Indicators with a mean score ≥8/10 for both importance and relevance were included in the final set. Overall, 108 QIs (70 process, 18 structural, 18 outcome, 1 patient engagement, and 1 covering multiple domains) were identified in 30 articles representing 23 agencies, organizations, and societies. Disease-specific QIs included myocardial infarction (n = 37), heart failure (n = 31), atrial fibrillation (n = 11), and cardiac rehabilitation (n = 1); general QIs represented about one-quarter (n = 28) of all measures. Fifteen QIs were selected for the final QI set: 7 process, 2 structural, and 6 outcome measures, including 6 general and 9 disease-specific measures. Outcome measures chosen to evaluate general CICU performance included overall CICU mortality, length of stay, and readmission rate. Numerous QIs relevant to the CICU have been recommended by a variety of organizations. The indicators chosen by the cardiac intensivist-administrators could serve as a basis for future efforts to develop a standardized set of quality measures for the CICU.


[Pathomorphology of the kidneys in hemolytic-uremic syndrome in children].
The kidneys were studied morphologically in 64 cases of hemolytic-uremic syndrome (HUS) in children aged from 17 days to 4 years. Three variants of the kidney alterations are distinguished depending on the duration of acute renal failure (ARF): early, late and delayed alterations. Early alterations were observed if ARF lasted for 1-10 days and were characterized by glomerular thrombotic microangiopathy with disturbance of the nephron proximal part function resultant from microcirculatory damage. Late kidney alterations (11-20 days of ARF) consisted of glomerular thrombi fibrinolysis, development of mesangial cell proliferation and mesangiolysis, proximal tubular epithelium regeneration and the appearance of nephrohydrosis resulting from the distal nephron part obstruction by hyalin cylinders. Delayed alterations (21-62 days of ARF) manifested with predominant atrophic and sclerotic processes. The first variant corresponded to the acute course of HUS while the two last variants to the subacute one.


Components study on gastroprotective effect and holistic mechanism of the herbal pair Alpinia officinarum - Cyperus rotundus based on spectrum-effect relationship and integrated transcriptome and metabolome analyses.
The herbal pair Alpinia officinarum-Cyperus rotundus (HPAC) has an extended history of use in the treatment of gastric ulcers, and its curative effect is definite. To explore the material basis and holistic mechanism of HPAC on ethanol-induced gastric ulcers. Three chemometrics, GRA, OPLS, and BCA, were used to construct the spectrum-effect relationship between the HPLC fingerprints of HPAC extracts and the bioactivity indices (cell viability; the levels of TNF-α, IL-6, COX-2, and PGE2; and wound healing rate) against GES-1 cell damage to screen the bioactive ingredients. The bioactive components were isolated and validated in vitro. Simultaneously, the effects of HPAC with concentrated bioactive ingredients was tested on ethanol-induced gastric ulcers in vivo, and the mechanism was investigated using transcriptomics and metabolomics. The mechanism was further validated by Western blotting. Finally, the contents of the main components of HPAC were determined before and after compatibility. Twelve bioactive components were screened, and the structures of nine compounds were confirmed. An in vitro verification test showed that DPHA and galangin could protect GES-1 cells from injury, and that their content increased after compatibility. The CH2Cl2 fraction of HPAC (HP-CH2Cl2) can protect mice from ethanol-induced gastric mucosal injury by reducing hemorrhage and decreasing inflammatory cell infiltration. Western blot analysis indicated that this fraction may up-regulate TRPV1 protein and down-regulate PI3K and AKT proteins. DPHA and galangin may be the bioactive components against ethanol-induced GES-1 cell injury. HP-CH2Cl2 may exert gastroprotective effects by regulating PI3K, AKT and TRPV1 proteins.


Is there any evidence for immunologically mediated or immunologically modifiable early pregnancy failure?
Human reproduction is an inefficient process. There is a high rate of loss of early pregnancies, often before the mother (or physician) knows she is pregnant. Genetic abnormalities can explain much of the wastage, but can it explain all of the failures? As embryos bear paternal and embryonic antigens foreign to the maternal immune system, could some otherwise normal embryos be "rejected"? Critical review of existing data. Otherwise normal embryos can fail prior to implantation, at implantation, in the periimplantation period as occult/chemical pregnancies, and as clinically evident miscarriages. The maternal immune system and its products (e.g., cytokines) can have innocent bystander effects, and a good case for direct recognition and "rejection" can also be made. The tools needed for accurate clinical diagnosis of such situations require further development and validation. Deliberate modification of the maternal host defence system can improve the chance of success, but the best evidence for efficacy of immunotherapeutic interventions is the situation of recurrent spontaneous abortions, which constitutes only a small percentage of losses. There is also evidence of clinical efficacy for several types of treatment to improve implantation and early pregnancy success.


Measuring the Cost of the Patient-Centered Medical Home: A Cost-Accounting Approach.
To explore the cost for individual practices to become more patient-centered, we inventoried and calculated the cost of costly activities involved in implementing the Patient-Centered Medical Home (PCMH) as defined by the National Committee for Quality Assurance. There were 3 key findings. The cost of each PCMH-related clinical activity can be classified in 1 of 3 major categories. Cost offsets can be used to defray part of the cost recognition. The cost of PCMH transformation varied by practice with no clear level or pattern of costs. Our study suggests that small- and medium-sized practices may experience difficulty with the financial burden of PCMH recognition.


Preparation and performance of hydroxypropyl methylcellulose esters of substituted succinates for in vitro supersaturation of a crystalline hydrophobic drug.
We prepared hydroxypropyl methylcellulose (HPMC) esters of substituted succinates and examined their performance for improving the aqueous solubility of crystalline hydrophobic drugs in spray-dried dispersions (SDDs). From one HPMC, we synthesized five HPMC esters using various monosubstituted succinic anhydrides. These HPMC esters along with a commercial HPMC acetate succinate (HPMCAS) were spray-dried from solutions with phenytoin. The SDDs with different matrices at 10 wt % loading had very similar bulk properties with a minimal amount of detectable crystalline phenytoin as revealed by scanning electron microscopy (SEM), powder X-ray diffraction (powder XRD), and differential scanning calorimetry (DSC). In solution, while the SDD with HPMCAS was very effective at achieving high levels of phenytoin supersaturation initially, it was not competent at maintaining such supersaturation due to the rapid crystallization of the dissolved phenytoin. Alternatively, SDDs with several synthesized HPMC esters of substituted succinates not only achieved rather high initial supersaturation but also maintained high concentrations for extended time (i.e., 1.5 h and longer). Such maintenance was largely ascribed to the inhibition of phenytoin nucleation. Structure-property relationships were established, and the most successful systems contained a high degree of substitution and a combination of a thioether with neighboring weak electron-withdrawing groups in the substituted succinic anhydrides. The effective maintenance of supersaturated solutions was only found in SDDs with rather low drug loadings, which indicates the significance of sufficiently high concentrations of polymer additives in the dissolution media.


Pollen tubes of Nicotiana alata express two genes from different beta-glucan synthase families.
The walls deposited by growing pollen tubes contain two types of beta-glucan, the (1,3)-beta-glucan callose and the (1,4)-beta-glucan cellulose, as well as various alpha-linked pectic polysaccharides. Pollen tubes of Nicotiana alata Link et Otto, an ornamental tobacco, were therefore used to identify genes potentially encoding catalytic subunits of the callose synthase and cellulose synthase enzymes. Reverse transcriptase-polymerase chain reactions (RT-PCR) with pollen-tube RNA and primers designed to conserved regions of bacterial and plant cellulose synthase (CesA) genes amplified a fragment that corresponded to an abundantly expressed cellulose-synthase-like gene named NaCslD1. A fragment from a true CesA gene (NaCesA1) was also amplified, but corresponding cDNAs could not be identified in a pollen-tube library, consistent with the very low level of expression of the NaCesA1 gene. RT-PCR with pollen-tube RNA and primers designed to regions conserved between the fungal FKS genes [that encode (1,3)-beta-glucan synthases] and their presumed plant homologs (the Gsl or glucan-synthase-like genes) amplified a fragment that corresponded to an abundantly expressed gene named NaGsl1. A second Gsl gene detected by RT-PCR (NaGsl2) was expressed at low levels in immature floral organs. The structure of full-length cDNAs of NaCslD1, NaCesA1, and NaGsl1 are presented. Both NaCslD1 and NaGsl1 are predominantly expressed in the male gametophyte (developing and mature pollen and growing pollen tubes), and we propose that they encode the catalytic subunits of two beta-glucan synthases involved in pollen-tube wall synthesis. Different beta-glucans deposited in one cell type may therefore be synthesized by enzymes from different gene families.


Identification of Sec36p, Sec37p, and Sec38p: components of yeast complex that contains Sec34p and Sec35p.
The Saccharomyces cerevisiae proteins Sec34p and Sec35p are components of a large cytosolic complex involved in protein transport through the secretory pathway. Characterization of a new secretion mutant led us to identify SEC36, which encodes a new component of this complex. Sec36p binds to Sec34p and Sec35p, and mutation of SEC36 disrupts the complex, as determined by gel filtration. Missense mutations of SEC36 are lethal with mutations in COPI subunits, indicating a functional connection between the Sec34p/sec35p complex and the COPI vesicle coat. Affinity purification of proteins that bind to Sec35p-myc allowed identification of two additional proteins in the complex. We call these two conserved proteins Sec37p and Sec38p. Disruption of either SEC37 or SEC38 affects the size of the complex that contains Sec34p and Sec35p. We also examined COD4, COD5, and DOR1, three genes recently reported to encode proteins that bind to Sec35p. Each of the eight genes that encode components of the Sec34p/sec35p complex was tested for its contribution to cell growth, protein transport, and the integrity of the complex. These tests indicate two general types of subunits: Sec34p, Sec35p, Sec36p, and Sec38p seem to form the essential core of a complex to which Sec37p, Cod4p, Cod5p, and Dor1p seem to be peripherally attached.


Use of the the combination labetalol plus chlorthalidone in essential arterial hypertension therapy.
Eleven patients suffering from WHO stage II essential arterial hypertension were treated with the combination of labetalol plus chlorthalidone in the reciprocal ratio of 10:1 in the form of once-a-day administration in the morning. The dosage was individualized on the basis of hypotensive response and side effects until an acceptable blood pressure was achieved. The duration of the treatment was 45 days with blood pressure measurements after 7, 15, and 45 days. By the end of the first week of therapy significant reductions had already been obtained in systolic arterial pressure (down from 181 +/- 24 to 157 +/- 9 mmHg, P less than 0.01) and diastolic arterial pressure (down from 110 +/- 7 to 96 +/- 7 mmHg, P less than 0.01), reductions which were also maintained over time. The mean dosage at the time of the final measurement was 150 mg (136 mg labetalol and 14 mg chlorthalidone). In no case were orthostatic hypotension phenomena observed, and the side effects proved modest without requiring suspension of the therapy. Because of the simplification of the therapy, patient compliance was good.


[Hospitalization demand for medical specialties in a health area with 200,000 inhabitants].
To assess the hospitalisation requirements for medical specialties (MS), admissions to a district hospital were studied during two subsequent years. Of the 196,800 inhabitants, 3% (6,089) were admitted which resulted in 7,841 admissions during this period. Four hundred and forty two different main diagnosis were performed but only 14 led to 50% of admissions and these same entities were also involved in any of the diagnosis associated (comorbidity). Mean age of the total group was 61 +/- 18 years being women patients three years older than men. More men than women were admitted. Pathology complexity was usually due to comorbidity, rather than a specific entity. Therefore we consider that the relationship between primary, general medicine and specialty attention should be reviewed and the involvement of MS in district hospitals should be revised so that the service may be efficient as well as improving cost-effectiveness ratio and thus achieving a remarkable economical saving.


Violence against children perpetrated by peers: A cross-sectional school-based survey in Uganda.
Violence against children by peers is a global public health problem. We aimed to assess factors associated with peer violence victimization among primary school children in Uganda. We conducted multilevel multivariable logistic regression analyses of cross-sectional data from 3706 primary students in 42 Ugandan primary schools. Among primary school students, 29% and 34% had ever experienced physical and emotional violence perpetrated by their peers, respectively. Factors strongly associated with both physical and emotional violence were similar and overlapping, and included exposure to interparental violence, having an attitude supportive of violence against children from school staff, not living with biological parents, working for payment, and higher SDQ score. However, we found that younger age, sharing sleeping area with an adult and achieving a higher educational performance score, were specifically associated with physical violence. On the other hand, being female, walking to school, reporting disability and eating one meal on the previous day, were particularly associated with emotional violence. Interventions to reduce peer violence should focus on family contexts, school environments and those with poor socio-economic status may need extra support.


Use of electrocochleography for assessing endolymphatic hydrops in patients with Lyme disease and Ménière's disease.
From an otological standpoint, Lyme disease can manifest itself as Ménière's disease both clinically and electrophysiologically. The aim of this study was to describe the findings of routine clinical, auditory and vestibular tests in patients with Ménière's and Lyme disease and to use electrocochleography (ECoG) to assess the presence of endolymphatic hydrops (EH) in both diseases. Transtympanic ECoG was performed in 91 patients with Ménière's disease and in 11 patients with confirmed Lyme disease. In both diseases the majority of patients had more than one complaint. There was one case with isolated hearing loss in the Lyme disease group. Typical clinical manifestations of Ménière's disease (vertigo, sensorineural hearing loss and tinnitus) were found in 6/11 patients (54.5%) in the Lyme disease group. The ECoG results indicated that there were 65/91 patients (71.4%) with Ménière's disease and 5 patients (45.5%) with Lyme disease who presented with EH. No statistically significant difference was found between the incidence of different symptoms of Ménière's and Lyme disease. On the basis of these results, patients with Lyme disease should undergo careful examination and investigation, especially in endemic regions. The presence of EH does not exclude the presence of infection with borreliosis as a cause of Ménière's disease-like symptoms.


Changing the tumor microenvironment: new strategies for immunotherapy.
Solid tumors are composed of malignant cells surrounded by a tumor-conditioned stroma that contains extracellular matrix and a variety of nonmalignant populations, including myeloid cells, lymphocytes, fibroblasts, and endothelial cells. These stromal elements form a local immunoregulatory network that must be overcome to achieve eradication of established tumors by immunotherapy. On March 21-22, 2012, a symposium was held in Pamplona, Spain, to share the recent advances regarding the molecules and cells that create and sustain this immune-hostile tumor microenvironment. Excellent targets for immunotherapeutic intervention were identified, and a number of therapeutic strategies under translation from mouse to human were presented.


CYP2U1, a novel human thymus- and brain-specific cytochrome P450, catalyzes omega- and (omega-1)-hydroxylation of fatty acids.
Long chain fatty acids have recently emerged as critical signaling molecules in neuronal, cardiovascular, and renal processes, yet little is presently known about the precise mechanisms controlling their tissue distribution and bioactivation. We have identified a novel cytochrome P450, CYP2U1, which may play an important role in modulating the arachidonic acid signaling pathway. Northern blot and real-time PCR analysis demonstrated that CYP2U1 transcripts were most abundant in the thymus and the brain (cerebellum), indicating a specific physiological role for CYP2U1 in these tissues. Recombinant human CYP2U1 protein, expressed in baculovirus-infected Sf9 insect cells, was found to metabolize arachidonic acid exclusively to two region-specific products as determined by liquid chromatography-mass spectrometry. These metabolites were identified as 19- and 20-hydroxy-modified arachidonic acids by liquid chromatography-tandem mass spectrometry analysis. In addition to omega/omega-1 hydroxylation of arachidonic acid, CYP2U1 protein also catalyzed the hydroxylation of structurally related long chain fatty acid (docosahexaenoic acid) but not fatty acids such as lauric acid or linoleic acid. This is the first report of the cloning and functional expression of a new human member of P450 family 2, CYP2U1, which metabolizes long chain fatty acids. Based on the ability of CYP2U1 to generate bioactive eicosanoid derivatives, we postulate that CYP2U1 plays an important physiological role in fatty acid signaling processes in both cerebellum and thymus.


Mouse and human ornithine decarboxylase genes. Methylation polymorphism and amplification.
With the use of the isoschizomeric restriction endonucleases HpaII and MspI, we found that mouse tumour ornithine decarboxylase (ODC; EC 184.108.40.206) genes are extensively methylated. ODC genes in L1210 mouse leukaemia cells were apparently more methylated than in Ehrlich ascites carcinoma, as revealed by the use of HpaII endonuclease, yet the digestion of genomic DNA isolated from these two murine tumour cell lines with MspI, which cleaves at a CCGG sequence, also with internally methylated cytosine, resulted in an apparently identical restriction pattern. It is possible that the amplification of ODC genes in Ehrlich ascites-carcinoma cells in response to 2-difluoromethylornithine (DFMO) was associated with hypomethylation, or that less-methylated genes were amplified. A human myeloma (Sultan) cell line only revealed three separate hybridization signals when cleaved with HpaII. One of these signals was amplified under the pressure of DFMO. When cleaved with MspI, these three HpaII fragments disappeared and were replaced by a double signal of 2.3-2.4 kilobase-pairs (kbp) in size. The amplified ODC sequences in the Sultan myeloma cell line apparently originated from chromosome 2, as indicated by a unique hybridization signal in a 5.8 kbp HindIII fragment specific for the human ODC locus on chromosome 2. A comparison of different human cells, the Sultan myeloma, a lymphocytic B-cell leukaemia (Ball), normal mononuclear leucocytes and leucocytes obtained from leukaemia patients, revealed interesting differences in the methylation of ODC genes. The use of two restriction endonucleases (HpaII and CfoI), the cleavage site for both of which contains a CG sequence and which only cleave when cytosine is unmethylated, indicated that ODC genes in the lymphocytic leukaemia cells were much less methylated than those in the normal leucocytes or in the Sultan cells.


New Developments in Traveler's Diarrhea.
Traveler's diarrhea (TD) is a crucial area for research, as it affects millions of tourists each year and creates a large economic burden. More than 60% of TD cases are caused by a variety of bacterial enteropathogens: diarrhea-producing Escherichia coli, Shigella, Campylobacter, Salmonella, Aeromonas, Plesiomonas, and noncholera Vibrios. Noroviruses are also an important cause of morbidity among travelers. Recent studies have identified host genetic risk factors associated with susceptibility to pathogen-specific TD. Prevention strategies should be emphasized, as all individuals with TD experience approximately 24 hours of disability and 5-10% experience chronic functional bowel disease. Poorly absorbed rifaximin provides protection for trips lasting 2 weeks or less. TD vaccines are also currently in development. All individuals planning trips to developing regions should be armed with 1 of the 3 agents that have shown efficacy for self-treatment of TD: ciprofloxacin (or levofloxacin), rifaximin, or azithromycin, depending upon the location of the trip. Although global epidemiologic changes in etiologic agents as well as antibiotic resistance patterns have been better understood recently, changes should be expected during the next decade due to new prevention and treatment approaches.


Occurrence of mycoplasmas in semen samples of birds of prey.
Mycoplasmas are well-known pathogens in a variety of animals. In poultry it is known that some species can be transmitted by semen and infect the uterus of females. As the prevalence of mycoplasmas in birds of prey is very high and artificial insemination is a commonly used technique for reproduction, the possibility of transmission Mycoplasma spp. by contaminated semen in birds of prey was investigated. Isolation of mycoplasmas was possible in five out of 32 (15.6%) semen samples of different bird of prey species. Two additional semen samples were positive for mycoplasma DNA using a Mycoplasma-genus-specific polymerase chain reaction. The isolation of mycoplasmas from a testicular sample indicates the testis as the possible source of contamination. Sequencing of large parts (>90%) of the 16S rRNA gene of the isolated mycoplasmas suggests that all isolates belong to the same species. Alignment of the sequenced products with the 16S rRNA gene of Mycoplasma species in GenBank demonstrated a similarity of 97% to Mycoplasma verecundum, but serological testing by immunobinding assay failed to identify it as such. It is recommended that the semen of donor birds of prey is examined for mycoplasmas before its use in artificial insemination.


Association Analysis Identifies New Loci for Resistance to Chinese Yr26-Virulent Races of the Stripe Rust Pathogen in a Diverse Panel of Wheat Germplasm.
Stripe rust caused by Puccinia striiformis f. sp. tritici (Pst) is one of the most destructive fungal diseases of wheat worldwide. The expanding Yr26-virulent Pst race (V26) group overcomes almost all currently deployed resistance genes in China and has continued to accumulate new virulence. Investigating the genetic architecture of stripe rust resistance in common wheat is an important basis for a successful utilization of resistance in breeding programs. A panel of 410 exotic wheat germplasms was used for characterizing new stripe rust resistance loci. This panel was genotyped using high-density wheat 660K single-nucleotide polymorphism (SNP) array, and phenotypic evaluation of seedlings for stripe rust resistance was performed using multiple Pst races. Thirty-five loci conferring resistance were identified through genome-wide association mapping, and explained phenotypic variances ranged from 53 to 75%. Of these, 14 were colocated in the proximity of the known loci, including cataloged Yr genes Yr9, Yr10, Yr26, Yr33, Yr47, Yr56, Yr57, Yr64, Yr67, Yr72, and Yr81 and three temporarily designated as YrCen, YrNP63, and YrRC detected in our quantitative trait locus (QTL) mapping studies. Seven of them (Yr9, Yr10, Yr24/26, Yr81, YrCEN, YrNP63, and YrRC) were confirmed by molecular detection or genetic analysis. New loci that were identified to be different from reported Yr genes need further confirmation. Nine QTL with significantly large phenotypic effect on resistance to all tested races were considered as major loci for effective resistance. The identified loci enrich our stripe rust resistance gene pool, and the linked SNPs should be useful for marker-assisted selection in breeding programs.


Comparison of neurological and functional outcomes after administration of granulocyte-colony-stimulating factor in motor-complete versus motor-incomplete postrehabilitated, chronic spinal cord injuries: a phase I/II study.
Granulocyte-colony-stimulating factor (G-CSF) is a major growth factor in the activation and differentiation of granulocytes. This cytokine has been widely and safely employed in different disease conditions over many years. The administration of the growth factors in spinal cord injury (SCI) has been reported elsewhere; here we have tried to see the effect of SCI severity on the neurological outcomes after neuroprotective treatment for SCI with G-CSF. Seventy-four consecutive patients with SCI of at least 6 months' duration, with stable neurological status in the last 3 months, having informed consent for the treatment were included in the study. All the patients had undergone at least 3 months of standard rehabilitation. Patients were assessed by the American Spinal Injury Association (ASIA) scale, Spinal Cord Independence Measure (SCIM) III, and International Association of Neurorestoratology-Spinal Cord Injury Functional Rating Scale (IANR-SCIFRS) just before intervention and periodically until 6 months after subcutaneous administration of 5 µg/kg per day of G-CSF for 7 consecutive days. Multiple linear regression models were performed for statistical evaluation of lesion completeness and level of injury on changes in ASIA motor, light touch, pinprick, IANR-SCIFRS, and SCIM III scores, as a phase I/II comparative study. The study consisted of 52 motor-complete and 22 motor-incomplete SCI patients. There was no significant difference regarding age and sex, chronicity, and level of SCI between the two groups. Motor-incomplete patients had significantly more improvement in ASIA motor score compared to the motor-complete patients (7.68 scores, p < 0.001); also they had significant improvement in light touch (6.42 scores, p = 0.003) and pinprick sensory scores (4.89 scores, p = 0.011). Therefore, G-CSF administration in motor-incomplete SCIs is associated with significantly higher motor improvement, and also the higher the initial ASIA Impairment Scale (AIS) grade, the less would be the final AIS change, and incomplete cases are more welcome into the future studies. This manuscript is published as part of the International Association of Neurorestoratology (IANR) special issue of Cell Transplantation.


Dense particles and slow sedimenting particles produced by ultraviolet irradiation of poliovirus.
Low doses of u.v. radiation rapidly inactivate poliovirus, and the virus is progressively converted into dense particles (DPs) of buoyant density 1.44 g/ml in CsCl. The DPs are structurally and antigenically related to standard virus (N-antigen), i.e. they are indistinguishable from virus in their RNA and protein content and in their sedimentation properties. Furthermore, there is no difference in reactivity of the structural proteins of virus and DPs with the monofunctional reagent [3H]N-succinimidyl propionate (3H-NSP). However, DPs differ from virus in that their capsids are permeable to several ions, and they can be degraded by RNase and protease. Increasing the radiation dose causes a successive transformation of DPs into 105S slow-sedimenting particles (SSPs). The SSPs are antigenically related to 76S artificial empty capsids (AECs) or H-antigen, but they differ physically and structurally from them. The SSPs have a higher S value than AECs and contain all the capsid proteins, including VP4, and the RNA, both of these macromolecules being absent from AECs. It is concluded, therefore, that transformation from N- to H-antigenicity by u.v. radiation does not require release of RNA and VP4. Conversion of virus particles to SSPs correlates with altered reactivity of VP2 and to a lesser extent VP1 and VP3, with the monofunctional reagent 3H-NSP.


Management of postmenopausal osteoporosis and the prevention of fractures.
Postmenopausal osteoporosis affects millions of women, being estrogen deficiency the key factor in the pathogenesis of involutional osteoporosis. Fracture prevention is one of the public health priorities worldwide. Different treatments for osteoporosis are available. The various options are aimed to maintain bone health and decrease the risk of fractures. The majority of these drugs are antiresorptive agents, i.e., drugs that lower bone turnover, inhibiting osteoclastic bone resorption. Dietary sources of calcium intake and vitamin D are ideal, while pharmachological supplements should be used if diet alone cannot provide the recommended daily intake. Bisphosphonates are first-line therapy for patients with established osteoporosis at high risk of fracture. Some serious, but rare, adverse events have been associated with their long-term administration. The monoclonal antibody to RANKL, named denosumab, administered as a 60-mg subcutaneous injection every 6 months, is a valuable option for the treatment of postmenopausal osteoporosis in women at increased or high risk of fractures, who are unable to take other osteoporosis treatments. Teriparatide (PTH 1-34) is the only available osteoanabolic drugs for osteoporosis treatment at present. Its use is limited to severe osteoporosis because of the high cost of the treatment. In climacteric women, in different stages of menopausal transition, and beyond, hormone replacement therapy at different doses (HRT) rapidly normalizes turnover, preventing and/or treating osteoporosis. HRT is able to preserve and even increase BMD at all skeletal sites, leading to a significant reduction in vertebral and non-vertebral fractures. Selective estrogen modulators (SERMs) as raloxifene and bazedoxifene reduce bone turnover and maintains or increases vertebral and femoral BMDs in comparison to placebo and reduces the risk of vertebral and new vertebral fractures, in high risk women. The combination of a SERM with an estrogen has been defined as tissue selective estrogen complex (TSEC). The bazedoxifene with conjugated estrogen is able to reduce climacteric symptoms, reducing bone turnover and preserving BMD. Studies investigating the actions of phytoestrogens on BMD or bone turnover are largely contradictory, making them inconclusive. At the present time, phytoestrogens cannot be recommended for postmenopausal osteoporosis. In conclusion, the use of HRT for osteoporosis prevention is based on biology, epidemiology, animal and preclinical data, observational studies and randomized, clinical trials. Osteoporosis prevention can actually be considered as a major additional effect in climacteric women who use HRT for treatment of climacteric symptoms. Bone protection is one of the major benefits of HRT. The possibility that low dose HRT or TSEC causes a decrease in fracture risk is not demonstrated but the scientific evidence is compelling. Conversely, established osteoporosis, often occurring in elderly women, can better be treated with specific treatments, such as bisphosphonates or, in more severe and selected cases, anabolic agents (teriparatide).


Biological responses to PDGF-BB versus PDGF-DD in human mesangial cells.
Platelet-derived growth factor (PDGF)-BB and PDGF-DD mediate mesangial cell proliferation in vitro and in vivo. While PDGF-BB is a ligand for the PDGF alpha- and beta-receptor chains, PDGF-DD binds more selectively to the beta-chain, suggesting potential differences in the biological activities. Signal transduction and regulation of gene expression induced by PDGF-BB and -DD were compared in primary human mesangial cells (HMCs), which expressed PDGF alpha- and beta-receptor subunits. The growth factor concentrations used were chosen based on their equipotency in inducing HMCs proliferation and binding to the betabeta-receptor. Both growth factors, albeit at different concentrations induced phosphorylation and activation of extracellular signal-regulated kinase 1 (ERK1) and ERK2. In addition, PDGFs led to the phosphorylation and activation of signal transducers and activators of transcription 1 (STAT1) and STAT3. HMCs proliferation induced by either PDGF-BB or -DD could be blocked by signal transduction inhibitors of the mitogen-activated protein kinase-, Janus kinase (JAK)/STAT-, or phosphatidyl-inositol 3-kinase pathways. Using a gene chip array and subsequent verification by real-time reverse transcriptase (RT)-polymerase chain reaction, we found that in HMC genes for matrix metalloproteinase 13 (MMP-13) and MMP-14 and, to a low extent, cytochrome B5 and cathepsin L were exclusively regulated by PDGF-BB, whereas no exclusive gene regulation was detected by PDGF-DD. However, at the protein level, both MMP-13 and -14 were equally induced by PDGF-BB and -DD. PDGF-BB and -DD effect similar biological responses in HMCs albeit at different potencies. Rare apparently differential gene regulation did not result in different protein expression, suggesting that in HMCs both PDGFs exert their biological activity almost exclusively via the PDGF beta-receptor.


Antibacterial and antioxidant activities of a novel biosynthesized selenium nanoparticles using Rosa roxburghii extract and chitosan: Preparation, characterization, properties, and mechanisms.
Developing efficient and safe antibacterial agents to inhibit pathogens including Physalospora piricola and Staphylococcus aureus is of great importance. Herein, a novel compound composed of Rosa roxburghii procyanidin, chitosan and selenium nanoparticle (RC-SeNP) was bio-synthesized, with the average diameter and zeta potential being 84.56 nm and -25.60 mV, respectively. The inhibition diameter of the RC-SeNP against P. piricola and S. aureus reached 18.67 mm and 13.13 mm, and the maximum scavenging activity against DPPH and ABTS reached 96.02% and 98.92%, respectively. Moreover, the RC-SeNP completely inhibited the propagation P. piricola and S. aureus on actual apples, suggesting excellent in vivo antimicrobial capacity. The transcriptome analysis and electron microscope observation indicated that the antibacterial activity would be attributed to adhering to and crack the cell walls as well as damage the cytomembrane and nucleus. Moreover, the RC-SeNP effectively maintained the vitamin C, total acid, and water contents of red bayberry, demonstrating potential application for fruit preservation. At last, the RC-SeNP showed no cell toxicity and trace selenium residual dose (0.03 mg/kg on apple, 0.12 mg/kg on red bayberry). This study would enlighten future development on novel nano-bioantibacterial agents for sustainable agriculture.


Identification of genes regulated by lipids from seaweed Susabinori (Pyropia yezoensis) involved in the improvement of hepatic steatosis: Insights from RNA-Seq analysis in obese db/db mice.
Hepatic steatosis is an early stage in the progression of non-alcoholic fatty liver disease (NAFLD) and can lead to the development of non-alcoholic steatohepatitis (NASH), a major cause of liver-related morbidity and mortality. Identification of dietary components that can alleviate hepatic steatosis is crucial for developing effective therapeutic strategies for NAFLD. Recently, we demonstrated the impact of lipids extracted from the marine red alga Susabinori (Pyropia yezoensis) in a murine model of type 2-diabete (db/db). We found that Susabinori lipids (SNL), abundant in eicosapentaenoic acid (EPA)-containing polar lipids, protected against obesity-induced hepatic steatosis in db/db mice. To understand the specific genes or biological pathways underlying the effects of SNL, we conducted RNA-Seq analysis of the hepatic transcriptome. By performing comparative analysis of differentially expressed genes between normal mice and db/db mice consuming a control diet, as well as SNL-fed db/db mice, we identified the 15 SNL-dependent up-regulated genes that were down-regulated in db/db mice but up-regulated by SNL feeding. Gene ontology and pathway analysis on these 15 genes demonstrated a significant association with the metabolisms of arachidonic acid (AA) and linoleic acid (LA). Furthermore, we observed alterations in the expression levels of monoacylglycerol lipase (Magl) and fatty acid-binding protein 4 (Fabp4) in the SNL-fed db/db mice, both of which are implicated in AA and LA metabolism. Additionally, the livers of SNL-fed db/db mice exhibited reduced levels of AA and LA, but a high accumulation of EPA. In conclusion, the SNL diet might affect the metabolisms of AA and LA, which contribute to the improvement of hepatic steatosis. Our findings provide insights into the molecular mechanisms underlying the beneficial effects of SNL.


Increased lymphocyte apoptosis in mouse models of colitis upon ABT-737 treatment is dependent upon BIM expression.
Exaggerated activation of lymphocytes contributes to the pathogenesis of inflammatory bowel disease (IBD). Medical therapies are linked to the BCL-2 family-mediated apoptosis. Imbalance in BCL-2 family proteins may cause failure in therapeutic responses. We investigated the role of BCL-2 inhibitor ABT-737 for lymphocyte apoptosis in mice under inflammatory conditions. B.6129P2-interleukin (IL)-10(tm1Cgn) /J (IL-10(-/-) ) weighing 25-30 g with ongoing colitis were used. Fifty mg/kg/day ABT-737 was injected intraperitoneally (i.p.). Haematological analyses were performed with an ADVIA 2120 flow cytometer and mass cytometry with a CyTOF 2. Following i.p. administration, ABT-737 was detected in both spontaneous and acute colitis in peripheral blood (PBL) and colon tissue. Treatment led to lymphopenia. CD4(+) CD44(+) CD62L(+) central memory and CD8(+) , CD44(+) CD62L(-) central memory T cells were decreased in PBL upon ABT-737 compared to vehicle-receiving controls. Increased apoptosis upon ABT-737 was determined in blood lymphocytes, splenocytes and Peyer's patches and was accompanied by a decrease in TNF and IL-1B. ABT-737 positively altered the colonic mucosa and ameliorated inflammation, as shown by colonoscopy, histology and colon length. A decreased BIM/BCL-2 ratio or absence of BIM in both Bim(-) (/) (-) and Il10(-) (/) (-) × Bim(-) (/) (-) impeded the protective effect of ABT-737. The BIM/BCL-2 ratio decreased with age and during the course of treatment. Thus, long-term treatment resulted in adapted TNF levels and macroscopic mucosal damage. ABT-737 was efficacious in diminishing lymphocytes and ameliorating colitis in a BIM-dependent manner. Regulation of inappropriate survival of lymphocytes by ABT-737 may provide a therapeutic strategy in IBD.


Heroin uncertainties: Exploring users' perceptions of fentanyl-adulterated and -substituted 'heroin'.
The US is experiencing an unprecedented opioid overdose epidemic fostered in recent years by regional contamination of the heroin supply with the fentanyl family of synthetic opioids. Since 2011 opioid-related overdose deaths in the East Coast state of Massachusetts have more than tripled, with 75% of the 1374 deaths with an available toxicology positive for fentanyl. Fentanyl is 30-50X more potent than heroin and its presence makes heroin use more unpredictable. A rapid ethnographic assessment was undertaken to understand the perceptions and experiences of people who inject drugs sold as 'heroin' and to observe the drugs and their use. A team of ethnographers conducted research in northeast Massachusetts and Nashua, New Hampshire in June 2016, performing (n=38) qualitative interviews with persons who use heroin. (1) The composition and appearance of heroin changed in the last four years; (2) heroin is cheaper and more widely available than before; and (3) heroin 'types' have proliferated with several products being sold as 'heroin'. These consisted of two types of heroin (alone), fentanyl (alone), and heroin-fentanyl combinations. In the absence of available toxicological information on retail-level heroin, our research noted a hierarchy of fentanyl discernment methods, with embodied effects considered most reliable in determining fentanyl's presence, followed by taste, solution appearance and powder color. This paper presents a new 'heroin' typology based on users' reports. Massachusetts' heroin has new appearances and is widely adulterated by fentanyl. Persons who use heroin are trying to discern the substances sold as heroin and their preferences for each form vary. The heroin typology presented is inexact but can be validated by correlating users' discernment with drug toxicological testing. If validated, this typology would be a valuable harm reduction tool. Further research on adaptations to heroin adulteration could reduce risks of using heroin and synthetic opioid combinations.


Diarrhoeagenic Escherichia coli and Escherichia albertii in Brazil: pathotypes and serotypes over a 6-year period of surveillance.
Diarrhoeagenic Escherichia coli (DEC) is a leading cause of infectious diarrhoea worldwide. In recent years, Escherichia albertii has also been implicated as a cause of human enteric diseases. This study describes the occurrence of E. coli pathotypes and serotypes associated with enteric illness and haemolytic uremic syndrome (HUS) isolated in Brazil from 2011 to 2016. Pathotypes isolated included enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC) and Shiga toxin-producing E. coli (STEC). PCR of stool enrichments for DEC pathotypes was employed, and E. albertii was also sought. O:H serotyping was performed on all DEC isolates. A total of 683 DEC and 10 E. albertii strains were isolated from 5047 clinical samples. The frequencies of DEC pathotypes were 52.6% (359/683) for EPEC, 32.5% for EAEC, 6.3% for ETEC, 4.4% for EIEC and 4.2% for STEC. DEC strains occurred in patients from 3 months to 96 years old, but EPEC, EAEC and STEC were most prevalent among children. Both typical and atypical isolates of EPEC and EAEC were recovered and presented great serotype heterogeneity. HUS cases were only associated with STEC serotype O157:H7. Two E. albertii isolates belonged to serogroup O113 and one had the stx2f gene. The higher prevalence of atypical EPEC in relation to EAEC in community-acquired diarrhoea in Brazil suggests a shift in the trend of DEC pathotypes circulation as previously EAEC predominated. This is the first report of E. albertii isolation from active surveillance. These results highlight the need of continuing DEC and E. albertii surveillance, as a mean to detect changes in the pattern of pathotypes and serotypes circulation and provide useful information for intervention and control strategies.


Parents with severe mental illness and their children: the need for human services integration.
This article presents the findings and recommendations of a statewide interagency task force on parents with mental illness who have young children. Based on testimony from consumers, providers, and advocates, the task force concluded that this is a substantial and neglected public policy issue requiring an intergovernmental, services integration approach. Recommendations were made in the areas of services, policies and procedures, and service coordination.


BRCA1 is an essential regulator of heart function and survival following myocardial infarction.
The tumour suppressor BRCA1 is mutated in familial breast and ovarian cancer but its role in protecting other tissues from DNA damage has not been explored. Here we show a new role for BRCA1 as a gatekeeper of cardiac function and survival. In mice, loss of BRCA1 in cardiomyocytes results in adverse cardiac remodelling, poor ventricular function and higher mortality in response to ischaemic or genotoxic stress. Mechanistically, loss of cardiomyocyte BRCA1 results in impaired DNA double-strand break repair and activated p53-mediated pro-apoptotic signalling culminating in increased cardiomyocyte apoptosis, whereas deletion of the p53 gene rescues BRCA1-deficient mice from cardiac failure. In human adult and fetal cardiac tissues, ischaemia induces double-strand breaks and upregulates BRCA1 expression. These data reveal BRCA1 as a novel and essential adaptive response molecule shielding cardiomyocytes from DNA damage, apoptosis and heart dysfunction. BRCA1 mutation carriers, in addition to risk of breast and ovarian cancer, may be at a previously unrecognized risk of cardiac failure.


Combined Structure- and Ligand-Based Approach for the Identification of Inhibitors of AcrAB-TolC in Escherichia coli.
The inhibition of efflux pumps is a promising approach to combating multidrug-resistant bacteria. We have developed a combined structure- and ligand-based model, using OpenEye software, for the identification of inhibitors of AcrB, the inner membrane protein component of the AcrAB-TolC efflux pump in Escherichia coli. From a database of 1391 FDA-approved drugs, 23 compounds were selected to test for efflux inhibition in E. coli. Seven compounds, including ivacaftor (25), butenafine (19), naftifine (27), pimozide (30), thioridazine (35), trifluoperazine (37), and meloxicam (26), enhanced the activity of at least one antimicrobial substrate and inhibited the efflux pump-mediated removal of the substrate Nile Red from cells. Ivacaftor (25) inhibited efflux dose dependently, had no effect on an E. coli strain with genomic deletion of the gene encoding AcrB, and did not damage the bacterial outer membrane. In the presence of a sub-minimum inhibitory concentration (MIC) of the outer membrane permeabilizer colistin, ivacaftor at 1 μg/mL reduced the MICs of erythromycin and minocycline by 4- to 8-fold. The identification of seven potential AcrB inhibitors shows the merits of a combined structure- and ligand-based approach to virtual screening.


Inhibition of cancer cell growth by anti-Tn monoclonal antibody MLS128.
Tn-antigens are generally masked by covalently linked carbohydrates but are exposed in most primary and metastatic epithelial malignant tumors, providing sensitive markers for detection of carcinoma. Here, therapeutic potentials of tumor-associated carbohydrate antigen-specific antibodies were investigated. MLS128, an anti-Tn monoclonal antibody, binds to a carbohydrate epitope consisting of three consecutive Tn-antigens (GalNAcalpha-Ser/Thr). MLS128 treatment significantly inhibited colon and breast cancer cell growth. MLS128 bound to 110-210 kDa glycoproteins on the cell surface. MLS128 treatment caused down-regulation of insulin-like growth factor-I receptor and epidermal growth factor receptor in LS180 colon cancer cells, suggesting that MLS128-inhibited cancer cell growth is in part mediated by down-regulation of growth factor receptors. This study provides the first insights into the potential use of this particular type of anti-Tn antigen antibodies as a cancer therapeutic.


Comment on "epidermal growth factor receptor is essential for toll-like receptor 3 signaling".
Epidermal growth factor receptor (EGFR) and mammalian target of rapamycin (mTOR) play important roles in tumor growth, which has stimulated efforts toward the design of targeted cancer therapeutics that inhibit their function. A growing body of literature indicates that EGFR and mTOR are also essential to support a functional innate immune response. Hence, although combination therapies that block both EGFR and mTOR may have improved activity against tumors, they may also place patients at risk of fulminant infections. We discuss data supporting this hypothesis.


Emotions Evoked by Colors and Health Functionality Information of Colored Rice: A Cross-Cultural Study.
This study aimed to examine the emotional responses evoked by cooked colored rice and its health functionality information in both consumers who eat rice as a staple food and consumers who do not eat rice as a staple food. Specifically, Korean and American consumers were exposed to colored rice and its health functionality information and an emotion lexicon was generated and measured based on focus group interviews (FGI) and two online consumer surveys. In test 1, the emotions evoked by presentation of stimuli to Koreans (N = 10) and Americans (N = 10) were extracted through FGIs and the first online consumer survey (Koreans = 69; Americans = 68) and an emotion lexicon was generated. As a result, a total of 34 terms were confirmed. Test 2 was conducted during the second online consumer survey (capturing data from a total of 208 Koreans and 208 Americans), utilizing the terms generated in test 1. In this test, only the colors (CO) of colored rice were presented to one group, while colors and health functionality information (CO&H) were presented to the other group. The overall liking for stimuli in both countries was highly correlated with familiarity. Koreans showed significantly more familiarity and liking for CO of white and black CO rice, while Americans showed significantly more familiarity and liking for CO of white and yellow rice. Hierarchical cluster analysis was performed to categorize the emotion terms, and the emotion terms were sorted into the three clusters, "Positive", "Negative", and "New", for both countries. Under informed conditions, the emotions became more positive, and emotions in the "New" cluster were evoked in both countries. The current study employed a cross-cultural approach to assess consumers' emotional responses to colored rice and health functionality information. Our findings suggest that providing foods with preferred colors for each culture and providing sufficient information on the said foods will help to promote unfamiliar foods.


Nardostachys jatamansi extract prevents chronic restraint stress-induced learning and memory deficits in a radial arm maze task.
Nardostachys jatamansi is traditionally used in alternative medicine for treatment of neuropsychiatric disorders. We investigated the potential of N. jatamansi extract (NJE) in protecting against chronic stress-induced impairments in spatial learning and memory. The rats were exposed to 21 days of chronic restraint stress and simultaneously received 100 mg or 200 mg/kg body weight of NJE following which acquisition and retention of hippocampus-dependent spatial memory were tested in a partially-baited eight arm radial maze. Animals treated with 200 mg/kg body weight NJE had learning curves comparable to control unstressed animals, made significantly more correct choices (38%, P < 0.001), and fewer reference memory errors (53%, P < 0.01) on the eighth day of training compared to untreated stressed animals as well as stressed animals which received vehicle or a lower dose (100 mg/kg) of NJE. NJE-treated animals also made significantly higher correct choices (31%, P < 0.001) than untreated animals in a retention test 10 days after the training period. We propose that NJE has a protective effect of stress-induced impairments in hippocampus-dependent learning and memory behavior in rats.


Multi-level 3D genome organization deteriorates during breast cancer progression.
Breast cancer entails intricate alterations in genome organization and expression. However, how three-dimensional (3D) chromatin structure changes in the progression from a normal to a breast cancer malignant state remains unknown. To address this, we conducted an analysis combining Hi-C data with lamina-associated domains (LADs), epigenomic marks, and gene expression in an in vitro model of breast cancer progression. Our results reveal that while the fundamental properties of topologically associating domains (TADs) remain largely stable, significant changes occur in the organization of compartments and subcompartments. These changes are closely correlated with alterations in the expression of oncogenic genes. We also observe a restructuring of TAD-TAD interactions, coinciding with a loss of spatial compartmentalization and radial positioning of the 3D genome. Notably, we identify a previously unrecognized interchromosomal insertion event, wherein a locus on chromosome 8 housing the MYC oncogene is inserted into a highly active subcompartment on chromosome 10. This insertion leads to the formation of de novo enhancer contacts and activation of the oncogene, illustrating how structural variants can interact with the 3D genome to drive oncogenic states. In summary, our findings provide evidence for the degradation of genome organization at multiple scales during breast cancer progression revealing novel relationships between genome 3D structure and oncogenic processes.


Response of primary glioblastoma cells to therapy is patient specific and independent of cancer stem cell phenotype.
Glioblastoma multiforme (GBM) contains a population of cells that exhibit stem cell phenotypes. These cancer stem cells (CSCs) may be a source of therapeutic resistance, although support for this important concept is limited. We determined whether early-passage GBM CSCs respond differently than patient-matched, genotypically similar non-CSCs to clinically relevant single or serial doses of temozolomide (TMZ), radiation therapy (XRT), or alternating TMZ treatment and XRT, which is the standard of care for GBM patients. Despite the phenotypic differences, including the presence of stem cell markers and formation of intracranial tumors, the CSCs and matched non-CSCs were equally resistant to TMZ in a majority of patients, using 2 independent assays. TMZ response was consistent with methylated O(6)-DNA methylguanine-methyltransferase (MGMT) and MGMT protein levels in both culture types. In contrast, CSCs were unexpectedly more responsive to XRT compared with matched non-CSCs from 2 patients despite having relatively equal resistance to TMZ. However, for the majority of culture pairs from individual patients, responses in CSCs were indistinguishable from non-CSC cultures. In our patient-matched primary cultures, response to TMZ was tightly linked to the individual tumor's MGMT status and independent of their phenotypic differences. TMZ and XRT together revealed no additive benefit compared with monotherapy for either culture type, in contrast to the notion that the CSC population is more resistant to XRT. If the tumor cell response in vitro mirrors therapeutic response in larger patient cohorts, these rapid assays in primary cultures could allow -empirical selection of efficacious therapeutic agents on a patient-specific basis.


High urinary dopa and low urinary dopamine-to-dopa ratio in salt-sensitive hypertension.
Dopamine in urine is derived substantially from renal uptake and decarboxylation of 3,4-dihydroxyphenylalanine (dopa), and increases in excretion of dopa normally parallel increases in excretion of dopamine during salt loading. Since patients with salt-sensitive hypertension may have decreased urinary excretion of dopamine during dietary salt loading, the present study was designed to evaluate the response of dopa to salt loading. Sixteen inpatients with normal-renin essential hypertension ate a constant metabolic diet containing 9 mmol/day sodium for 7 days, followed by the same diet but containing 249 mmol/day sodium for 7 days. Salt sensitivity was defined as an increase in mean arterial pressure of 8 mm Hg between the diets; on this basis, nine patients were salt-sensitive and seven, salt-resistant. The rate of urinary dopa excretion was significantly higher in the salt-sensitive patients throughout the study (mean rates 132 +/- 13 nmol/day in the salt-sensitive group and 78 +/- 9 nmol/day in the salt-resistant group for the 14 days of observation, p less than 0.01). When dietary sodium intake was increased to 249 mmol/day, urinary dopa excretion increased significantly more in salt-sensitive patients than salt-resistant patients. At the end of the high salt diet, dopamine excretion was significantly attenuated in the salt-sensitive patients, despite higher rates of dopa excretion. Thus, the urinary ratio of dopamine to dopa was decreased in salt-sensitive patients, regardless of salt intake.(ABSTRACT TRUNCATED AT 250 WORDS)


Single-Photon Emission from Organic Dye Molecules Adsorbed on a Quantum Dot via Energy Transfer.
Single-photon emissions from individual emitters are crucial in fundamental science and quantum information technologies. Multichromophoric systems, comprising multiple dyes, can exhibit single-photon emissions through efficient annihilation between the excited states; however, exploring this phenomenon in complex systems remains a challenge. In this study, we investigated the photon statistics of emissions from multiple perylene bisimide (PBI) dyes adsorbed onto the surface of CdSe/ZnS quantum dots (QDs). When multiple PBIs were simultaneously excited by both direct excitation and energy transfer from the QD, multiphoton emissions from the PBIs were observed. Conversely, when the QDs were selectively excited, multiple PBIs exhibiting single-photon emission through energy transfer from the QDs to the PBIs were found. These results highlight the intriguing interplay between multichromophoric systems and QDs, offering valuable insights into the development of efficient single-photon sources in quantum information technologies.


The antiphospholipid antibody syndrome in the emergency department setting--livedo reticularis and recurrent venous thrombosis.
We present the case of a 26-year-old man with an exacerbation of apparent chronic asthma with chronic peripheral vascular disease due to recurrent venous thrombosis. Localized livedo reticularis, new cutaneous infarctions, severe venous insufficiency, thrombocytopenia, renal failure, and cerebral supratentorial dysfunction were noted. During hospital admission, antibodies to phospholipids in high titer were present by three different testing methods. Renal biopsy demonstrated significant renal vasculature abnormalities characteristic of hemolytic endovasculopathy, and magnetic resonance imaging showed multiple cerebral infarctions. This case exemplifies the spectrum of presentations and management of the primary antiphospholipid antibody syndrome. The clue to its presence in this patient was the livedo reticularis rash, a cutaneous marker for this syndrome that was evident in the emergency department.


Shapes of carcinogenic benz[alpha]anthracenes: an x-ray crystal structure analysis of 12-methylbenz[alpha]anthracene.
The crystal structure of the moderately active carcinogen 12-methylbenz[alpha]anthracene (12-MBA) has been determined by application of direct methods to X-ray single-crystal diffraction data. Least-squares refinement to a residual R = 0.09 over 929 independent reflections enabled carbon positions to be established with apparent e.s.d.s. of atomic coordinates about 0.008 A. Deviation from planarity is exemplified by the 15.5 degrees inclination of the benz ring (A) to the anthracene nucleus and by the 0.89 A distance of the methyl carbon out of the best plane through the whole benzanthracene nucleus. Comparison with the structure of the highly carcinogenic 7,12-dimethylbenz[alpha]anthracene (7,12-DMBA), and with the recently solved structures of the weak carcinogen 1-MBA and the extremely weak carcinogen 1,12-DMBA, shows a close similarity in the anthracene parts; in 1-MBA, and 1,12-DMBA, the phenanthrenic K-region bond is close to 1.34 A and the M-region bond about 1.38 A. In 12-MBA, overcrowding in the 'bay' region causes the central anthracene ring C and the benz ring A each to be bent about 10 degrees in opposite directions from the phenanthrenic B ring, much as in 1-MBA and 7,12-DMBA, but less than in 1,12-DMBA; the 12-methyl carbon lies about the same distance (0.55 A) above the anthracene plane in 12-MBA as in 1,12-MBA and 7,12-DMBA.


Mutant p53(R175H) upregulates Twist1 expression and promotes epithelial-mesenchymal transition in immortalized prostate cells.
A mutation within one allele of the p53 tumor suppressor gene can inactivate the remaining wild-type allele in a dominant-negative manner and in some cases can exert an additional oncogenic activity, known as mutant p53 'gain of function' (GOF). To study the role of p53 mutations in prostate cancer and to discriminate between the dominant-negative effect and the GOF activity of mutant p53, we measured, using microarrays, the expression profiles of three immortalized prostate epithelial cultures expressing wild-type, inactivated p53 or mutated p53. Analysis of these gene expression profiles showed that both inactivated p53 and p53(R175H) mutant expression resulted in the upregulation of cell cycle progression genes. A second group, which was upregulated exclusively by mutant p53(R175H), was predominantly enriched in developmental genes. This group of genes included the Twist1, a regulator of metastasis and epithelial-mesenchymal transition (EMT). Twist1 levels were also elevated in metastatic prostate cancer-derived cell line DU145, in immortalized lung fibroblasts and in a subset of lung cancer samples, all in a mutant p53-dependent manner. p53(R175H) mutant bearing immortalized epithelial cells showed typical features of EMT, such as higher expression of mesenchymal markers, lower expression of epithelial markers and enhanced invasive properties in vitro. The mechanism by which p53(R175H) mutant induces Twist1 expression involves alleviation of the epigenetic repression. Our data suggest that Twist1 expression might be upregulated following p53 mutation in cancer cells.


Can online support groups address psychological morbidity of cancer patients? An artificial intelligence based investigation of prostate cancer trajectories.
Online Cancer Support Groups (OCSG) are becoming an increasingly vital source of information, experiences and empowerment for patients with cancer. Despite significant contributions to physical, psychological and emotional wellbeing of patients, OCSG are yet to be formally recognised and used in multidisciplinary cancer support programs. This study highlights the opportunity of using Artificial Intelligence (AI) in OCSG to address psychological morbidity, with supporting empirical evidence from prostate cancer (PCa) patients. A validated framework of AI techniques and Natural Language Processing (NLP) methods, was used to investigate PCa patient activities based on conversations in ten international OCSG (18,496 patients- 277,805 conversations). The specific focus was on activities that indicate psychological morbidity; the reasons for joining OCSG, deep emotions and the variation from joining through to milestones in the cancer trajectory. Comparative analyses were conducted using t-tests, One-way ANOVA and Tukey-Kramer post-hoc analysis. PCa patients joined OCSG at four key phases of psychological distress; diagnosis, treatment, side-effects, and recurrence, the majority group was 'treatment' (61.72%). The four groups varied in expression of the intense emotional burden of cancer. The 'side-effects' group expressed increased negative emotions during the first month compared to other groups (p<0.01). A comparison of pre-treatment vs post-treatment emotions showed that joining pre-treatment had significantly lower negative emotions after 12-months compared to post-treatment (p<0.05). Long-term deep emotion analysis reveals that all groups except 'recurrence' improved in emotional wellbeing. This is the first empirical study of psychological morbidity and deep emotions expressed by men with a new diagnosis of cancer, using AI. PCa patients joining pre-treatment had improved emotions, and long-term participation in OCSG led to an increase in emotional wellbeing, indicating a decrease in psychological distress. It is opportune to further investigate AI in OCSG for early psychological intervention as an adjunct to conventional intervention programs.


Juxtaglomerular cell complex in the regulation of renal salt excretion.
Luminal NaCl concentration at the macula densa (MD) has the two established effects of regulating glomerular arteriolar resistance and renin secretion. Tubuloglomerular feedback (TGF), the inverse relationship between MD NaCl concentration and glomerular filtration rate (GFR), stabilizes distal salt delivery and thereby NaCl excretion in response to random perturbations unrelated to changes in body salt balance. Control of vasomotor tone by TGF is exerted primarily by NaCl transport-dependent changes in local adenosine concentrations. During long-lasting perturbations of MD NaCl concentration, control of renin secretion becomes the dominant function of the MD. The potentially maladaptive effect of TGF under chronic conditions is prevented by TGF adaptations, permitting adjustments in GFR to occur. TGF adaptation is mechanistically coupled to the end point targeted by chronic deviations in MD NaCl, the rate of local and systemic angiotensin II generation. MD control of renin secretion is the result of the coordinated action of local mediators that include nitric oxide synthase (NOS) and cyclooxygenase (COX) products. Thus vascular smooth muscle cell activation during high MD transport and granular cell activation during low MD transport is achieved by different extracellular mediators. The coordinated regulation of NOS I and COX-2 expression in MD cells and of renin expression in granular cells suggests that control of juxtaglomerular regulation of gene transcription or mRNA metabolism may be another consequence of a chronic alteration in MD NaCl concentration.


Immune response to rotavirus polypeptides after vaccination with heterologous rotavirus vaccines (RIT 4237, RRV-1).
The antibody response to individual rotavirus polypeptides after vaccination with live attenuated heterologous rotavirus vaccines was studied with a radioimmunoprecipitation assay (RIPA). Following vaccination with bovine rotavirus strain RIT 4237 and rhesus monkey rotavirus strain RRV-1, an antibody response was demonstrated mainly against the inner capsid polypeptides VP2 and VP6 and to a lesser extent against VP3/VP4. There was no qualitative difference between the vaccines in the antibody response; both vaccines induced antibodies against the same polypeptides. Quantitatively the antibody response to RRV-1 was somewhat stronger. All prevaccination sera contained by RIPA low levels of rotavirus antibodies. The effect of these antibodies on the 'take' of the vaccines is discussed.


Quantification of the amount of mobile components in intact stratum corneum with natural-abundance 13C solid-state NMR.
The outermost layer of the skin is the stratum corneum (SC), which is mainly comprised of solid proteins and lipids. Minor amounts of mobile proteins and lipids are crucial for the macroscopic properties of the SC, including softness, elasticity and barrier function. Still this minor number of mobile components are not well characterized in terms of structure or amount. Conventional quantitative direct polarization (Q-DP) 13C solid-state NMR gives signal amplitudes proportional to concentrations, but fails to quantify the SC mobile components because of spectral overlap with the overwhelming signals from the solids. Spectral editing with the INEPT scheme suppresses the signals from solids, but also modulates the amplitudes of the mobile components depending on their values of the transverse relaxation times T2, scalar couplings JCH, and number of covalently bound hydrogens nH. This study describes a quantitative INEPT (Q-INEPT) method relying on systematic variation of the INEPT timing variables to estimate T2, JCH, nH, and amplitude for each of the resolved resonances from the mobile components. Q-INEPT is validated with a series of model systems containing molecules with different hydrophobicity and dynamics. For selected systems where Q-DP is applicable, the results of Q-INEPT and Q-DP are similar with respect to the linearity and uncertainty of the obtained molar ratios. Utilizing a reference compound with known concentration, we quantify the concentrations of mobile lipids and proteins within the mainly solid SC. By melting all lipids at high temperature, we obtain the total lipid concentration. These Q-INEPT results are the first steps towards a quantitative understanding of the relations between mobile component concentrations and SC macroscopic properties.


Building Additional Passageways in Polyamide Membranes with Hydrostable Metal Organic Frameworks To Recycle and Remove Organic Solutes from Various Solvents.
Membrane separation is a promising technology for extracting temperature-sensitive organic molecules from solvents. However, a lack of membrane materials that are permeable toward organic solvents yet highly selective curtails large-scale membrane applications. To overcome the trade-off between flux and selectivity, additional molecular transportation pathways are constructed in ultrathin polyamide membranes using highly hydrostable metal organic frameworks with diverse functional surface architectures. Additional passageways enhance water permeance by 84% (15.4 L m-2 h-1 bar-1) with nearly 100% rose bengal rejection and 97.6% azithromycin rejection, while showing excellent separation performance in ethyl acetate, ketones, and alcohols. These unique composite membranes remain stable in both aqueous and organic solvent environments. This immediately finds application in the purification of aqueous mixtures containing organic soluble compounds, such as antibiotics, during pharmaceutical manufacturing.


Do delays in radiology lead to breaches in the 4-hour rule?
To assess trends in medical imaging requests before and after the 4-hour rule commenced and to assess the imaging time component of emergency department (ED) length of stay (LOS). Retrospective analysis of ED patients and imaging requests 1 year prior to and 3 years after implementation of the 4-hour rule (April to December for 2011-2014) was performed at a single adult tertiary referral Level 1 trauma hospital with Level 6 ED. Logistic regression was used to evaluate trends in the number of ED patient presentations, patient triage categories, and imaging requests for these patients. The imaging component of the total ED LOS was compared for patients who met the 4-hour target and patients who did not. Compared to 2011 (before the 4-hour rule), ED presentations increased 4.74% in 2012, 12.7% in 2013, 21.28% in 2014 (p<0.01). Total imaging requests increased 23.05% in 2012, 48.04% in 2013, 60.77% in 2014 (p<0.01). For patients breaching the 4-hour rule, the mean time before radiology request was 2.4-2.8 hours; mean time from imaging request to completion was 1.2-1.3 hours; mean time from imaging completion to discharge from ED was the longest component of ED LOS (4.9-5.9 hours). There has been a significant increase in imaging requests, with a trend towards more CT and less radiography requests. Imaging requests for patients who breached the 4-hour target were made on average 2.4-2.8 hours after triage and average time after imaging in itself, exceeded 4 hours. Imaging is not likely a causative factor for patients breaching the 4-hour target.


Leukoreduction system chambers are a reliable cellular source for the manufacturing of T-cell therapeutics.
Following solid organ or hematopoietic cell transplantation, refractory opportunistic viral reactivations are a significant cause of morbidity and mortality but can effectively be controlled by virus-specific T-cell transfer. Among effective and safe strategies is the use of "third-party" (neither from the transplant donor nor recipient) virus-specific T cells that can be manufactured from healthy donors and used as "off-the-shelf" therapies. Leukoreduction system chambers (LRSCs), recovered after routine plateletpheresis, were evaluated as a potential source of peripheral blood mononuclear cells (PBMCs) for the manufacturing of clinical-scale virus-specific T cell. PBMCs from the same donors obtained either from LRSCs or peripheral blood were compared, focusing on T-cell function and phenotype as well as the potential to generate cytomegalovirus (CMV)-specific T-cell lines from both CMV seropositive and seronegative donors. PBMCs from both sources were comparable except for a transient downregulation of CD62L expression on freshly extracted PBMCs from LRSCs. Both nonspecific stimulation using anti-CD3/CD28 antibodies and CMV peptides revealed that LRSCs or blood T cells were equivalent in terms of expansion, differentiation, and function. Moreover, PBMCs from LRSCs can be used to generate autologous monocyte-derived dendritic cells to prime and expand CMV-specific T cells from seronegative donors. LRSCs are a reliable source of PBMCs for the generation of virus-specific T cells for immunotherapy. These findings have implications for the development of third-party therapeutic T-cell products from well-characterized blood product donors.


A betaine adduct of N-heterocyclic carbene and carbodiimide, an efficient ligand to produce ultra-small ruthenium nanoparticles.
The betaine adduct of N-heterocyclic carbene and carbodiimide (ICy·((p-tol))NCN) was found to be a very efficient ligand to prepare very small (1-1.3 nm) ruthenium nanoparticles (RuNPs). The coordination of the ligand on the metal surface takes place through the carbodiimide moiety. The resulting RuNPs led to decarbonylation of THF and showed size selectivity for styrene hydrogenation.


3D Reconstruction of Human Motion from Monocular Image Sequences.
This article tackles the problem of estimating non-rigid human 3D shape and motion from image sequences taken by uncalibrated cameras. Similar to other state-of-the-art solutions we factorize 2D observations in camera parameters, base poses and mixing coefficients. Existing methods require sufficient camera motion during the sequence to achieve a correct 3D reconstruction. To obtain convincing 3D reconstructions from arbitrary camera motion, our method is based on a-priorly trained base poses. We show that strong periodic assumptions on the coefficients can be used to define an efficient and accurate algorithm for estimating periodic motion such as walking patterns. For the extension to non-periodic motion we propose a novel regularization term based on temporal bone length constancy. In contrast to other works, the proposed method does not use a predefined skeleton or anthropometric constraints and can handle arbitrary camera motion. We achieve convincing 3D reconstructions, even under the influence of noise and occlusions. Multiple experiments based on a 3D error metric demonstrate the stability of the proposed method. Compared to other state-of-the-art methods our algorithm shows a significant improvement.


Actin-dependent endosomal receptor recycling.
Endosomes constitute major sorting compartments within the cell. There, a myriad of transmembrane proteins (cargoes) are delivered to the lysosome for degradation or retrieved from this fate and recycled through tubulo-vesicular transport carriers to different cellular destinations. Retrieval and recycling are orchestrated by multi-protein assemblies that include retromer and retriever, sorting nexins, and the Arp2/3 activating WASH complex. Fine-tuned control of actin polymerization on endosomes is fundamental for the retrieval and recycling of cargoes. Recent advances in the field have highlighted several roles that actin plays in this process including the binding to cargoes, stabilization of endosomal subdomains, generation of the remodeling forces required for the biogenesis of cargo-enriched transport carriers and short-range motility of the transport carriers.


Allergen-specific immunotherapy.
Specific immunotherapy was introduced for the treatment of grass pollen-induced hay fever in 1911. The treatment was soon extended to other pollens as well as perennial allergens, and to the treatment of bronchial asthma. Definitive studies of its efficacy for both rhinitis and asthma came only many decades later. Understanding gradually emerged of the underlying immunologic mechanisms that include the generation of regulatory T lymphocytes, immune deviation from allergen-specific Th2 to Th1 responses, and a shift in allergen-specific antibody production from immunoglobulin (Ig) E to IgG4. Along with understanding of the immune basis came an appreciation that immunotherapy modifies allergic disease expression, producing protection against disease progression and symptomatic improvement that persists for years after the treatment is discontinued. Recent new directions for immunotherapy include sublingual administration of inhalant allergens and use of the oral route to treat food allergy.


(1)H NMR metabolomics identification of markers of hypoxia-induced metabolic shifts in a breast cancer model system.
Hypoxia can promote invasive behavior in cancer cells and alters the response to therapeutic intervention as a result of changes in the expression many genes, including genes involved in intermediary metabolism. Although metabolomics technologies are capable of simultaneously measuring a wide range of metabolites in an untargeted manner, these methods have been relatively under utilized in the study of cancer cell responses to hypoxia. Thus, (1)H NMR metabolomics was used to examine the effects of hypoxia in the MDA-MB-231 human breast cancer cell line, both in vitro and in vivo. Cell cultures were compared with respect to their metabolic responses during growth under either hypoxic (1% O(2)) or normoxic conditions. Orthogonal partial least squares discriminant analysis (OPLS-DA) was used to identify a set of metabolites that were responsive to hypoxia. Via intracardiac administration, MDA-MB-231 cells were also used to generate widespread metastatic disease in immuno-compromised mice. Serum metabolite analysis was conducted to compare animals with and without a large tumor burden. Intriguingly, using a cross-plot of the OPLS loadings, both the in vitro and in vivo samples yielded a subset of metabolites that were significantly altered by hypoxia. These included primarily energy metabolites and amino acids, indicative of known alterations in energy metabolism, and possibly protein synthesis or catabolism. The results suggest that the metabolite pattern identified might prove useful as a marker for intra-tumoral hypoxia.


Innate Immune System Activation, Inflammation and Corneal Wound Healing.
Corneal wounds resulting from injury, surgeries, or other intrusions not only cause pain, but also can predispose an individual to infection. While some inflammation may be beneficial to protect against microbial infection of wounds, the inflammatory process, if excessive, may delay corneal wound healing. An examination of the literature on the effect of inflammation on corneal wound healing suggests that manipulations that result in reductions in severe or chronic inflammation lead to better outcomes in terms of corneal clarity, thickness, and healing. However, some acute inflammation is necessary to allow efficient bacterial and fungal clearance and prevent corneal infection. This inflammation can be triggered by microbial components that activate the innate immune system through toll-like receptor (TLR) pathways. In particular, TLR2 and TLR4 activation leads to pro-inflammatory nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) activation. Similarly, endogenous molecules released from disrupted cells, known as damage-associated molecular patterns (DAMPs), can also activate TLR2, TLR4 and NFκB, with the resultant inflammation worsening the outcome of corneal wound healing. In sterile keratitis without infection, inflammation can occur though TLRs to impact corneal wound healing and reduce corneal transparency. This review demonstrates the need for acute inflammation to prevent pathogenic infiltration, while supporting the idea that a reduction in chronic and/or excessive inflammation will allow for improved wound healing.


[Our experience with revision total knee arthroplasty].
In the period from 1990 and June 2003, 885 total knee replacements were performed at the orthopedic ward of the Ceske Budejovice Hospital. Of these, 19 (2.14 %) patients underwent revision surgery; in addition, 25 patients who had had primary surgery in other hospitals were operated on. Of these 44 patients, 25 were followed up and evaluated. The aim of the study was to evaluate the pre-operative treatment including examination for bacterial infection by cultivation, the selection of an optimal procedure (one- or two-stage operation, surgical approach and implantation technique) and postoperative therapy. The 25 evaluated patients were followed up for an average of 34.5 months (range, 6-109 months) after reimplantation. Loosening occurred in most of the commonly used types of primary implants. The group comprised five men and 19 women; the average age at the time of reoperation was 70 years (range, 51-78 years). Ten patients had repeat surgery on the left and 14 on the right knee at an average of 43.5 months (range, 4-120 months) after primary surgery. Fourteen patients were treated by one-stage and 10 patients by two-stage surgery. The Genesis system (Smith Nephew) was used in 13 patients, Sigma PFC revision implant (Johnson and Johnson) in eight, Walter-Motorlet implant (reuse of the primary implant at the time when a revision system was not available) in one and external fixator in five patients. Revision arthroplasty was indicated on the basis of clinical symptoms and X-ray, scintigraphic and biochemical (CRP, WBC, FW) examinations. The use of recently adopted methods (procalcitonin, orosomucoid, alpha-1-antitrypsin, beta-2- macroglobulin, ceruloplasmin, PCR and PET) was not evaluated because of short-term applications. Patients in whom infection or colonization of the implant was suspected were treated by two-stage reimplantation, using a canalized spacer with a stem and a patellar pelota made of antibiotic-loaded cement. The average time between implant removal and reimplantation was 108 days (range, 60-244 days). Each removed implant was placed in a culture medium for 5 to 7 days. This resulted in a high occurrence of positive cultivation results even in the patients who, on the basis of previous examination, were first considered to have had aseptic loosening and had undergone one-stage surgery. All patients with positive tests received long-term antibiotic therapy, usually a combination of ciprofloxacin and rifampicin, according to the cultivation results. Out of 14 one-stage reimplantations (indicated for by the negative results of all laboratory examinations), implant colonization was recorded in five cases, with a coagulase-negative staphylococcus being the most frequent infectious agent. No recurrent infection was found after the long-term antibiotic course. One patient with the implant infected with Staphylococcus aureus underwent primary arthrodesis. Out of 10 two-stage reimplantations (in patients with positive laboratory tests), recurrent infection was found in two cases and was caused by a pathogen different from the original one. The patients were treated by arthrodesis. Good outcomes, defined as a functional total knee replacement free from infection at least 6 months after reimplantation, were achieved in 79 % of the patients. Better functional results were obtained by onestage surgery. Patients with concomitant rheumatoid arthritis had aseptic loosening more frequently, and patients with impaired immunological status, due to diabetes mellitus, cytostatic drug or corticosteroid administration, more often showed septic loosening. The 2.14 % loosening of total knee arthroplasty in our patients (19 out of 885) can be considered a good result. A functional joint was achieved in all patients (100 %) with aseptic loosening and in 69 % of those with infected or colonized implants. The results of routine biochemical tests and bacteriological cultivation did not allow us to distinguish aseptic from septic loosening with certainty. Therefore, we adopted new screening markers (PCR and PET) and a new method of microbiological examination of the removed implant and collected tissue. However, we could evaluate the role of these specialized tests only on the basis of literature data, since we had only short-term experience with them ourselves. Our results with the treatment of early loosening of total knee arthroplasty suggest that patients benefit more from the two-stage procedure. We strongly emphasize the employment of all possible means to prevent loosening, i. e., to use an appropriate surgical technique for primary implantation, to observe aseptic principles and to administer antibiotic therapy in conditions suspected of bacteremia. The shorter the interval between the onset of complaints and the reimplantation, the better results. Early loosening should be treated by two-stage surgery. Our method of bacteriological examination gives good results. Because of complexity of the problem, patients with a loose knee prosthesis should be referred to orthopedic departments with experienced and skilled surgical teams and high-quality examination facilities.With the observation of appropriate procedures, there is a great chance of achieving good results. Arthrodesis is still regarded as a justified "salvage" operation, particularly in cases with pre-operative findings of Staphylococcus aureus. Procedures for repeat surgery following the failure of a reimplanted joint have so far yielded doubtful results and still await further development.


Magnetic resonance imaging findings of shoulders in Parkinson's disease.
The aim of this study is to evaluate shoulder disturbances in Parkinson's disease (PD) patients using magnetic resonance imaging (MRI) which is the best tool in the demonstration of complex shoulder pathologies; and to determine probable relations between shoulder pathologies and PD clinical features. Twenty-eight PD patients with a total of 56 shoulders were used as the study group while 13 age-matched cases with 26 shoulders were used as the control group (CG) in the study. Both patients with PD and the CG underwent shoulder MRI. The Hoehn and Yahr (H&Y) disability scale and Unified Parkinson's Disease Rated Scale (UPDRS) were used to determine the severity of the disease. Our results showed that patients with full-thickness supraspinatus (SSP) tear have statistically significant higher UPDRS (P = 0.012), tremor (P = 0.023), rigidity (P = 0.023), and total (P = 0.002) scores. Mild group patients (P = 0.045) showed significantly higher frequency resting tremor and subcoracoid effusion than those of severe group patients (P = 0.002). Subcoracoid effusion was observed in patients with significantly higher UPDRS (P = 0.045) and rigidity (P = 0.022) scores. When the resting tremor and subcoracoid effusion groups were compared according to the severity of the resting tremor but not according to the H&Y, higher frequency of full-thickness tear in SSP tendon was detected in the group of resting tremor (P = 0.053). Longer duration of disease was also observed in patients with full-thickness SSP tear (P = 0.029) and acromioclavicular joint changes (P = 0.018). Higher UPDRS, tremor, rigidity and total scores and longer PD duration appear as the predisposing factors for the development of shoulder disturbances in PD in this study.


A prospective randomized multicenter trial comparing clinical outcomes of patients treated surgically with a static or dynamic implant for acute ankle syndesmosis rupture.
To compare the clinical and radiographic outcome after stabilization of an acute syndesmosis rupture with either a static implant (a 3.5-mm metallic screw through 4 cortices) or a dynamic device (TightRope; Arthrex). Multicenter randomized double-blind controlled trial. Study realized in 5 trauma centers (2 level 1 and 3 level 2) in 2 countries. Seventy subjects admitted for an acute ankle syndesmosis rupture entered the study and were randomized into 2 groups (dynamic fixation = 34 and static fixation = 36). The 2 groups were similar regarding demographic, social, and surgical data. Sixty-five patients (dynamic = 33 and static = 32) completed the study and were available for analysis. Syndesmosis fixation in the static group was realized with a 4 cortices 3.5-mm cortical screw (Synthes) and in the dynamic group with 1 TightRope (Arthrex). Standardized rehabilitation process for the 2 groups: no weight bearing in a cast for 6 weeks and then rehabilitation without protection. Olerud-Molander score. Subjects with dynamic fixation achieved better clinical performances as described with the Olerud-Molander scores at 3 (68.8 vs. 60.2, P = 0.067), 6 (84.2 vs. 76.8, P = 0.082), and 12 months (93.3 vs. 87.6, P = 0.046). We also observed higher American Orthopaedic Foot and Ankle Society scores at 3 months (78.6 vs. 70.6, P = 0.016), but these were not significant at 6 (87.1 vs. 83.8, P = 0.26) or 12 months (93.1 vs. 89.9, P = 0.26). Implant failure was higher in the screw group (36.1% vs. 0%, P < 0.05). Loss of reduction was observed in 4 cases in the static screw group (11.1% vs. 0%, P = 0.06). Dynamic fixation of acute ankle syndesmosis rupture with a dynamic device seems to result in better clinical and radiographic outcomes. The implant offers adequate syndesmotic stabilization without failure or loss of reduction, and the reoperation rate is significantly lower than with conventional screw fixation. Therapeutic Level II. See Instructions for Authors for a complete description of levels of evidence.


High-dimensional phenotyping of the peripheral immune response in community-acquired pneumonia.
Community-acquired pneumonia (CAP) represents a major health burden worldwide. Dysregulation of the immune response plays an important role in adverse outcomes in patients with CAP. We analyzed peripheral blood mononuclear cells by 36-color spectral flow cytometry in adult patients hospitalized for CAP (n=40), matched control subjects (n=31), and patients hospitalized for COVID-19 (n=35). We identified 86 immune cell metaclusters, 19 of which (22.1%) were differentially abundant in patients with CAP versus matched controls. The most notable differences involved classical monocyte metaclusters, which were more abundant in CAP and displayed phenotypic alterations reminiscent of immunosuppression, increased susceptibility to apoptosis, and enhanced expression of chemokine receptors. Expression profiles on classical monocytes, driven by CCR7 and CXCR5, divided patients with CAP into two clusters with a distinct inflammatory response and disease course. The peripheral immune response in patients with CAP was highly similar to that in patients with COVID-19, but increased CCR7 expression on classical monocytes was only present in CAP. CAP is associated with profound cellular changes in blood that mainly relate to classical monocytes and largely overlap with the immune response detected in COVID-19.


1.38-µm mode-locked Raman fiber laser pumped by semiconductor disk laser.
A mode-locked Raman fiber laser pumped by 1.3 µm semiconductor disk laser is demonstrated. Direct Watt-level core-pumping of the single-mode fiber Raman lasers and amplifiers with low-noise disk lasers is demonstrated to represent a highly practical solution as compared with conventional scheme using pumping by Raman wavelength convertors. Raman laser employing passive mode-locking by nonlinear polarization evolution in normal dispersion regime produces stable pedestal-free 1.97 ps pulses at 1.38 µm. Using semiconductor disk lasers capable of producing high power with diffraction-limited beam allows Raman gain to be obtained at virtually any wavelength of interest owing to spectral versatility of semiconductor gain materials and wafer-fusing technology.


Fast eigenspace decomposition of images of objects with variation in illumination and pose.
Many appearance-based classification problems such as principal component analysis, linear discriminant analysis, and locally preserving projections involve computing the principal components (eigenspace) of a large set of images. Although the online expense associated with appearance-based techniques is small, the offline computational burden becomes prohibitive for practical applications. This paper presents a method to reduce the expense of computing the eigenspace decomposition of a set of images when variations in both illumination and pose are present. In particular, it is shown that the set of images of an object under a wide range of illumination conditions and a fixed pose can be significantly reduced by projecting these data onto a few low-frequency spherical harmonics, producing a set of "harmonic images." It is then shown that the dimensionality of the set of harmonic images at different poses can be further reduced by utilizing the fast Fourier transform. An eigenspace decomposition is then applied in the spectral domain at a much lower dimension, thereby significantly reducing the computational expense. An analysis is also provided, showing that the principal eigenimages computed assuming a single illumination source are capable of recovering a significant amount of information from images of objects when multiple illumination sources exist.