Differences in mycorrhizal preferences between two tropical orchids.
Orchids parasitize their mycorrhizal fungi and are dependent on them for seed germination. Controversy reigns over how specific the mycorrhizal association is in tropical species. Although there is little experimental evidence to support any viewpoint, some variation is known to exist. We compared mycorrhizal specificity and performance in two phylogenetically related epiphytic orchids from Puerto Rico, Tolumnia variegata and Ionopsis utricularioides (Oncidiinae) by integrating two techniques: phylogenetic analysis of mycorrhizal fungi based on nuclear ribosomal internal transcribed spacer (ITS) sequences, and symbiotic seed germination experiments. Most of the mycorrhizal isolates from T. variegata fell into four different clades of Ceratobasidium, while most of those from I. utricularioides were restricted to a single clade of the same genus. Seeds of T. variegata germinated equally well with fungi from both T. variegata and I. utricularioides, but seeds of I. utricularioides germinated significantly better with its own isolates. Seeds of I. utricularioides germinated and developed faster than those of T. variegata. Both the molecular phylogeny and the seed germination experiments showed that T. variegata is a generalist in its association with fungal symbionts. In contrast, I. utricularioides is more specialized and more effective at exploiting a specific fungal clade. Our data are consistent with the theoretical trade-offs between specialized and generalized interactions.


Influencing factors of nursing caries.
This study was done to determine some of the effecting factors in the formation of nursing caries. For this purpose, 161 children with nursing caries, aged 2 to 5 years, were chosen. The control group was formed by 181 children with no caries. The ratio of giving up the habits in the group having nursing caries during the 12 months period was lower than the control group. The children falling asleep with the nursing caries bottle and a more incidence of nursing caries compared to those who did not. It was found that the type of carbohydrate liquid did not have a different effect in the formation of nursing caries. Commenced toothbrushing age did not show significant differences between those who developed nursing caries and those who did not. Even though the families had enough knowledge about the formation of caries, this was unable to stop the formation of the phenomenon.


The XXmotif web server for eXhaustive, weight matriX-based motif discovery in nucleotide sequences.
The discovery of regulatory motifs enriched in sets of DNA or RNA sequences is fundamental to the analysis of a great variety of functional genomics experiments. These motifs usually represent binding sites of proteins or non-coding RNAs, which are best described by position weight matrices (PWMs). We have recently developed XXmotif, a de novo motif discovery method that is able to directly optimize the statistical significance of PWMs. XXmotif can also score conservation and positional clustering of motifs. The XXmotif server provides (i) a list of significantly overrepresented motif PWMs with web logos and E-values; (ii) a graph with color-coded boxes indicating the positions of selected motifs in the input sequences; (iii) a histogram of the overall positional distribution for selected motifs and (iv) a page for each motif with all significant motif occurrences, their P-values for enrichment, conservation and localization, their sequence contexts and coordinates. Free access: http://xxmotif.genzentrum.lmu.de.


Hepatitis B and liver transplantation.
Liver transplantation is the treatment of choice for patients with liver failure secondary to chronic hepatitis B. However, liver transplantation is complicated by the risk of recurrent hepatitis B virus infection, which significantly impairs graft and patient survival. The main risk factor for the development of recurrent hepatitis B virus infection is the virus load at the time of transplantation. The development of antiviral medications, such as lamivudine and adefovir, and the implementation of effective prophylactic regimens using hepatitis B immune globulin have significantly improved the outcomes of hepatitis B after liver transplantation. However, current approaches continue to be hampered by the extremely high cost of treatment and the emergence of drug-resistant viral mutations. Ongoing studies are necessary to establish the most cost-effective approaches to prevent recurrent hepatitis B virus infection after liver transplantation.


Sexual polymorphisms of vomeronasal 1 receptor family gene expression in bulls, steers, and estrous and early luteal-phase heifers.
Vomeronasal 1 receptors (V1R) are a family of receptors for intraspecies chemosignals, including pheromones, and are expressed in the olfactory epithelium (OE) and vomeronasal organ (VO). Even in the well-studied rodents, it is unclear which members of the V1R family cause sexual polymorphisms, as there are numerous genes and it is difficult to quantify their expressions individually. Bovine species carry only 34 V1R homologs, and the OE and VOs are large enough to sample. Here, V1R expression was quantified in the OE and VOs of individual bovines. Based on the 34 gene sequences, we obtained a molecular dendrogram consisting of four clusters and six independent branches. Semi-quantitative RT-PCR was used to obtain gene expression profiles in the VOs and OE of 5 Japanese Black bulls, 5 steers, 7 estrous heifers and 6 early luteal-phase heifers. Ten genes showed significant between-group differences, and 22 showed high expression in VOs than in OE. The bulls showed higher expression of one gene more in OE and another in VOs (both P<0.05) than did steers; both genes belonged to the first cluster. No genes were expressed more abundantly in steers than in bulls. The estrous heifers showed higher expression of a gene of the second cluster in OE, and a gene of the third cluster in VOs (both P<0.05) than did early luteal-phase heifers. These results suggest V1R expression exhibits sexual polymorphisms in cattle.


The case for vaccines.
The case for vaccines is one which has been made through scientific advancement and public health implementation, resulting in one of the most significant historical achievements for mankind. This includes the elimination of endemic smallpox, polio, measles and rubella from the U.S. This exhilarating accomplishment was sobered with the threat of smallpox through biological attack following September 11, 2001. While the unthinkable return of that vaccine-preventable disease never materialized, other vaccine-preventable disease, such as pertussis, have markedly increased in many states because of never-established or waning immunity. Drivers of these current threats come both from the anti-vaccine movement, through legislative efforts to expand childhood immunization exemptions and the medical establishment itself through lack of immunization prioritization in adolescent and adult populations. Therefore, the case for vaccines needs to be made both externally and internally through sound science, sound logic and sound ethics. Most powerfully, however, the case for vaccines is told through stories of real people who have suffered or died from these preventable diseases.


The Application of a Low-temperature Physical Plasma Device Operating Under Atmospheric Pressure Leads to the Production of Toxic NO2.
Physical plasma is a mixture of reactive particles and electromagnetic radiation. Due to the antimicrobial, immunomodulatory, anti-inflammatory, wound-healing promoting, and antineoplastic effects of body tempered physical plasma under atmospheric pressure (cold atmospheric plasma: CAP), CAP therapy is increasingly becoming the focus of surgical and oncological disciplines. However, when applied in practice, a potential emission of harmful noxae such as toxic nitrogen oxides must be taken into account, which was investigated in the following study. MiniJet-R Ar CAP device was characterized with respect to NOX-specific spectra, ultraviolet radiation C (UVC) intensity in the range of 200-275 nm and the formation of NOX gases. Instrument-specific parameters such as gas flow, energy setting of the high-frequency generator, and flow rate of the carrier gas Ar were varied. To test the toxic properties of the NO2 concentrations formed by CAP, SK-OV-3 human ovarian cancer cells were incubated with different NO2 concentrations and cell growth was monitored for 120 h. The operation of MiniJet-R led to the formation of NO2 in the proximity of the CAP effluent. Synthesis of NO led to a NO-specific spectrum in the range of 100-275 nm, whereby UVC radiation produced reached intensities of up to 90 mW/m2 NO gas itself, however, was not detectable, as it was converted to NO2 rapidly. Cell culture incubation experiments demonstrated that NO2 in these concentration ranges had no influence on the cell growth of human cancer cells. Although no limit values were exceeded in the present study, the emission of high-energy UVC radiation and toxic NO2 is a risk factor with regard to the legal regulations on workplace protection (operator hazard) and the approval of medical devices (patient hazard). This is important for considerations regarding treatment frequency and duration. The growth inhibitory effect of CAP treatment on human cancer cells principally suggests a medical application of the MiniJet-R device, although more extensive studies will have to follow.


Assessment of the carotid artery Doppler flow time in patients with acute upper gastrointestinal bleeding.
Because of the subjectivity and ambiguity of the noninvasive measurements and limited use of invasive ones, there is an impending need for a real-time, fast, inexpensive, and reproducible noninvasive measurement method in acute upper gastrointestinal (GI) bleeding with active bleeding in emergency services. In this study, we aimed to evaluate the effect of bedside carotid artery flow time (CFT) measurement before and after the passive leg raising (PLR) maneuver on the determination of active bleeding in patients admitted to the emergency department (ED) with upper GI bleeding. This prospective case-control study was conducted in the ED of a training and research hospital with upper GI bleeding. Patients were placed in the supine position to perform bedside carotid Doppler ultrasonography before starting treatment. CFT, corrected CFT (CFTc), and carotid artery Doppler flow velocity were measured. After then performed PLR, the same parameters were measured again. A total of 94 patients, including 50 patients with GI bleeding and 44 healthy volunteers as control group were included in the study. CFT and CFTc were shorter in the patient group than the control group (P < 0.001, P = 0.004, respectively). After PLR, there were statistically significant differences in change in the CFT (ΔCFT) and change in the corrected CFT (ΔCFTc) between the groups (P = 0.001, P < 0.001). There were also statistically significant differences in ΔCFT and ΔCFT between the patients with active bleeding and the nonbleeding ones (P = 0.01, P = 0.005, respectively). Area under curve to detect active bleeding for ΔCFT and ΔCFTc were calculated as 0.801 (95% confidence interval [CI]: 0.65-0.95) and 0.778 (95% CI: 0.63-0.91), respectively. The corrected carotid Doppler flow time measurements in patients with GI bleeding at the time of presenting to the emergency department can be helpful to interpret the active bleeding.


Perceived control, academic performance and well-being of Ghanaian college students with disability.
Empirical evidence abounds showing the impact of perceived control on subjective well-being in several spheres of functioning, including academic performance. At tertiary institutions, such as the Kwame Nkrumah University of Science and Technology, Ghana, little is known about the needs of students with disabilities, as very few persons with disabilities attend institutions of higher learning. This study examined the relationship between perceptions of control and the academic and subjective well-being of students with disabilities. A total of 69 students with disabilities participated in this cross-sectional descriptive study. Using trusted control and subjective well-being scales, data were subject to descriptive analyses. Consistent with previous works, perceived control increased with increased subjective well-being, moderated by gender. In addition, forms of secondary control appeared to aid primary control in the tenacious pursuit of goals. However, neither perceived control nor self-esteem was predictive of academic performance. Limitations of sample size notwithstanding, the findings of the study can be considered provocative. Implications for clinical utility in facilitating context-specific interventions for this marginalised group are discussed. Replication with a larger sample size in other tertiary institutions is suggested for future work.


Ectopic gastric mucosa in the cervical esophagus.
This study describes the clinical presentation and management of ectopic gastric mucosa (EGM) in the cervical esophagus. This is a case report of a 53-year-old male who presented with left-sided odynophagia of 3 months' duration. Office examination, including flexible fiberoptic laryngoscopy, was unremarkable. Direct larynogoscopy and rigid esophagoscopy revealed a 2 x 1-cm fleshy, red, and raised lesion in the cervical esophagus. The remainder of the esophagus was normal. Histologic examination of a biopsy specimen from the lesion showed a normal gastric epithelium consistent with the diagnosis of ectopic gastric mucosa. He was treated with 3 months of proton pump inhibitors, which resulted in complete symptom resolution. Patient was then switched to H2-blockers and has been symptom-free for one year. The medical literature regarding EGM in the cervical esophagus is reviewed with respect to its incidence, pathogenesis, clinical course, and management. In conclusion, EGM can occur in the cervical esophagus, and odynophagia is the typical presenting symptom. Adenocarcinoma developing within random, nonsyndromic EGM of the cervical esophagus is exceedingly rare. The primary mode of treatment is medical therapy.


Properties of ultrasound-rapid MRI clinical diagnostic pathway in suspected pediatric appendicitis.
to determine diagnostic accuracy of an US-MRI clinical diagnostic pathway to detect appendicitis in the emergency department (ED). prospective cohort study of 624 previously healthy children 4-17 years old undergoing US for suspected appendicitis and clinical re-assessment. Children with non-diagnostic USs and persistent appendicitis concern/conclusive US-reassessment discrepancies underwent ultra-rapid MRI (US-MRI pathway), interpreted as positive, negative or non-diagnostic. Cases with missed appendicitis, negative appendectomies, and CT utilization were considered clinically diagnostically inaccurate. Primary outcome was the proportion of accurate diagnoses of appendicitis/lack thereof by the pathway. 150/624 (24%) children had appendicitis;255 USs (40.9%) were non-diagnostic. Of 139 US-MRI pathway children (after 117 non-diagnostic and 22 conclusive USs), 137 [98.6%; 95% CI 0.96-1.00] had clinically accurate outcomes (1 CT, 1 negative appendectomy): sensitivity 18/18 [100%], specificity 119/121 [98.3%], positive predictive value 18/20 [90.5%], negative predictive value 119/119 [100%]. MRI imaging accuracy was 134/139 (96.4%); 3 MRIs were non-diagnostic (no appendicitis). In the overall algorithm, 616/624 [98.7% (0.97-0.99)] patients had accurate outcomes: 147/150 (98.0%) appendicitis cases had confirmatory surgeries (3 CTs) and 469/474 (98.9%) appendicitis-negative children had no surgery/CT. this study demonstrated high clinical accuracy of the US-rapid-MRI pathway in suspected pediatric appendicitis after non-diagnostic US.


Tail-interacting protein TIP47 is a connector between Gag and Env and is required for Env incorporation into HIV-1 virions.
The presence of the envelope glycoprotein Env in HIV-1 virions is essential for infectivity. To date, the molecular mechanism by which Env is packaged into virions has been largely unknown. Here, we show that TIP47 (tail-interacting protein of 47 kDa), which has been shown to interact with Env, also binds the MA (matrix) domain of HIV-1 Gag protein and that these three proteins form a ternary complex. Mutations in Gag that abrogate interaction with TIP47 inhibit Env incorporation and virion infectivity as well as colocalization between Gag and Env. We also show that TIP47 silencing impairs Env incorporation and infectivity and abolishes coimmunoprecipitation of Gag with Env. In contrast, overexpression of TIP47 increases Env packaging. Last, we demonstrate that TIP47 can interact simultaneously with Env and Gag. Taken together, our results show that TIP47 is a cellular cofactor that plays an essential role in Env incorporation, allowing the encounter and the physical association between HIV-1 Gag and Env proteins during the viral assembly process.


[Celiac disease: etiology and susceptibility].
Celiac disease is the result of an immune response that only occurs in genetically predisposed individuals. Both innate and adquired immune responses have a role. In the acquired cellular immune response, the most important in the disease, T lymphocytes are activated by gliadin peptides presented by specific HLA-DQ molecules. However, even in HLA-DQ2 homozygotes, only 5% get the disease, whereas the concordance between identical twins is 80%. Studying candidate genes in more than 500 patients and controls we have found other susceptibility genes for the disease.


A synopsis of the prevalence rates and environmental risk factors for myopia.
The prevalence rates of myopia are higher in urban Asian cities such as Hong Kong and Singapore. One observation over the past few decades is that the prevalence rates of myopia have been rising and there is an epidemic of myopia in Asia. The age-old question of the roles of nature and nurture in this process remains unanswered. The strongest evidence for an environmental link to myopia is near work activity. Childhood exposure to night lighting has also been explored in different studies but the results have been mixed. Twin studies, segregation analysis and association studies have demonstrated that hereditary factors play an important role in myopia development. The exact nature and interplay of genetic and environmental factors is not known and data suggest that environmental factors may interact with genetic factors to increase the risks of developing myopia. Future research is needed to identify specific modifiable lifestyle factors and genetic markers for myopia. This will enable preventive measures such as health education to be instituted.


Type II NKT-TFH cells against Gaucher lipids regulate B-cell immunity and inflammation.
Chronic inflammation including B-cell activation is commonly observed in both inherited (Gaucher disease [GD]) and acquired disorders of lipid metabolism. However, the cellular mechanisms underlying B-cell activation in these settings remain to be elucidated. Here, we report that β-glucosylceramide 22:0 (βGL1-22) and glucosylsphingosine (LGL1), 2 major sphingolipids accumulated in GD, can be recognized by a distinct subset of CD1d-restricted human and murine type II natural killer T (NKT) cells. Human βGL1-22- and LGL1-reactive CD1d tetramer-positive T cells have a distinct T-cell receptor usage and genomic and cytokine profiles compared with the classical type I NKT cells. In contrast to type I NKT cells, βGL1-22- and LGL1-specific NKT cells constitutively express T-follicular helper (TFH) phenotype. Injection of these lipids leads to an increase in respective lipid-specific type II NKT cells in vivo and downstream induction of germinal center B cells, hypergammaglobulinemia, and production of antilipid antibodies. Human βGL1-22- and LGL1-specific NKT cells can provide efficient cognate help to B cells in vitro. Frequency of LGL1-specific T cells in GD mouse models and patients correlates with disease activity and therapeutic response. Our studies identify a novel type II NKT-mediated pathway for glucosphingolipid-mediated dysregulation of humoral immunity and increased risk of B-cell malignancy observed in metabolic lipid disorders.


Development of erythropoietin receptor-targeted drug delivery system against breast cancer using tamoxifen-loaded nanostructured lipid carriers.
Tamoxifen (TAM) has been used in the treatment of breast cancers and is supplemented with erythropoietin (EPO) to alleviate the cancer-related anemia. The purported deleterious effects caused by the use of EPO with chemotherapeutic agents in the treatment of cancer-related anemia vary across studies and remain controversial. The use of nanoparticles as a drug delivery system has the potential to improve the specificity of anticancer drugs. In this study, we simultaneously incorporated two pharmacological active ingredients in one nanocarrier to develop EPO-conjugated TAM-loaded lipid nanoparticles (EPO-TAMNLC), a targeted delivery system, to enhance the cytotoxic activity while reducing the side effects of the ingredients. The effect of temperature in modulating the thermodynamic parameters associated with the binding of EPO and TAMNLC was assessed using isothermal titration calorimetry, while the unfolding of EPO structure was determined using fluorescence-quenching approach. The association efficiency of EPO and TAMNLC was 55.43%. Unlike binding of albumin to TAMNLC, the binding of EPO to TAMNLC occurred through endothermic and entropy-driven reaction. The EPO-TAMNLC formulation was stable because of the hydrophobic interaction and the high free energy, suggesting the spontaneity of the interactions between EPO and TAMNLC. The EPO-TAMNLC enhanced the in vitro cytotoxicity of TAM to MCF-7 cells. The EPO surface-functionalized TAMNLC could sequentially deliver EPO and TAM as well as improving site-specific delivery of these therapeutic compounds.


The mutated S1-haplotype in sour cherry has an altered S-haplotype-specific F-box protein gene.
Gametophytic self-incompatibility (GSI) is an outcrossing mechanism in flowering plants that is genetically controlled by 2 separate genes located at the highly polymorphic S-locus, termed S-haplotype. This study characterizes a pollen part mutant of the S(1)-haplotype present in sour cherry (Rosaceae, Prunus cerasus L.) that contributes to the loss of GSI. Inheritance of S-haplotypes from reciprocal interspecific crosses between the self-compatible sour cherry cultivar Ujfehértói Fürtös carrying the mutated S(1)-haplotype (S(1)'S(4)S(d)S(null)) and the self-incompatible sweet cherry (Prunus avium L.) cultivars carrying the wild-type S(1)-haplotype revealed that the mutated S(1)-haplotype confers unilateral incompatibility with a functional pistil component and a nonfunctional pollen component. The altered sour cherry S(1)-haplotype pollen part mutant, termed S(1)', contains a 615-bp Ds-like element within the S(1)-haplotype-specific F-box protein gene (SFB(1)'). This insertion generates a premature in-frame stop codon that would result in a putative truncated SFB(1) containing only 75 of the 375 amino acids present in the wild-type SFB(1). S(1)' along with 2 other previously characterized Prunus S-haplotype mutants, S(f) and S(6m), illustrate that mobile element insertion is an evolutionary force contributing to the breakdown of GSI.


The Emerging Role of the IL-17B/IL-17RB Pathway in Cancer.
Among inflammatory mediators, a growing body of evidence emphasizes the contribution of the interleukin 17 (IL-17) cytokine family in malignant diseases. Besides IL-17A, the prototypic member of the IL-17 family, several experimental findings strongly support the role of the IL-17B/IL-17 receptor B (IL-17RB) pathway in tumorigenesis and resistance to anticancer therapies. In mouse models, IL-17B signaling through IL-17RB directly promotes cancer cell survival, proliferation, and migration, and induces resistance to conventional chemotherapeutic agents. Importantly, recent work by our and other laboratories showed that IL-17B signaling dramatically alters the tumor microenvironment by promoting chemokine and cytokine secretion which foster tumor progression. Moreover, the finding that elevated IL-17B is associated with poor prognosis in patients with pancreatic, gastric, lung, and breast cancer strengthens the results obtained in pre-clinical studies and highlights its clinical relevance. Here, we review the current understanding on the IL-17B/IL-17RB expression patterns and biological activities in cancer and highlight issues that remain to be addressed to better characterize IL-17B and its receptor as potential targets for enhancing the effectiveness of the existing cancer therapies.


The effect of F-actin on the binding and hydrolysis of guanine nucleotide by Dictyostelium elongation factor 1A.
Indirect evidence implicates actin as a cofactor in eukaryotic protein synthesis. The present study directly examines the effects of F-actin on the biochemical properties of eukaryotic elongation factor 1A (eEF1A, formerly EF1alpha), a major actin-binding protein. The basal mechanism of eEF1A alone is determined under physiological conditions with the critical finding that glycerol and guanine nucleotide are required to prevent protein aggregation and loss of enzymatic activity. The dissociation constants (Kd) for GDP and GTP are 2.5 microM and 0.6 microM, respectively, and the kcat of GTP hydrolysis is 1.0 x 10(-3) s-1. When eEF1A binds to F-actin, there is a 7-fold decrease in the affinity for guanine nucleotide and an increase of 35% in the rate of GTP hydrolysis. Based upon our results and the relevant cellular concentrations, the predominant form of cellular eEF1A is calculated to be GTP.eEF1A.F-actin. We conclude that F-actin does not significantly modulate the basal enzymatic properties of eEF1A; however, actin may still influence protein synthesis by sequestering GTP.eEF1A away from interactions with its known translational ligands, e.g. aminoacyl-tRNA and ribosomes.


Gonadal Maturation Changes Chemotaxis Behavior and Neural Processing in the Olfactory Circuit of Caenorhabditis elegans.
Many animal species change their behavior according to their stage of development. However, the mechanisms involved in translating their developmental stage into the modifications of the neuronal circuits that underlie these behavioral changes remain unknown. Here we show that Caenorhabditis elegans changes its olfactory preferences during development. Larvae exhibit a weak chemotactic response to the food-associated odor diacetyl, whereas adults exhibit a strong response. We show that germline loss, caused either by laser ablation of germline precursor cells or mutations, results in a diacetyl-specific chemotactic defect in adult animals. These results suggest that germline cells, which proliferate dramatically during the larval stages, enhance chemotaxis to diacetyl. Removal experiments of specific neurons suggested that AWA olfactory neurons and their downstream interneurons, AIA and AIB, are required for germline-dependent chemotactic enhancement. Calcium imaging in animals lacking germline cells indicates that the neural responses of AWA and AIB to diacetyl stimuli are decreased compared with animals with an intact germline. These changes in neural activities may at least partly explain the behavioral change of animals lacking germline cells. Furthermore, this germline-dependent chemotactic change depends on the transcription factor DAF-16/FOXO. We find that organismal behavior changes throughout development by integrating information about physiological status from internal tissues to modify a simple sensory circuit.


Age-related differences in the use of total shoulder arthroplasty over time: use and outcomes.
We assessed the age-related differences in the use of total shoulder arthroplasty (TSA) and outcomes, and associated time-trends using the United States Nationwide Inpatient Sample (NIS) between 1998 and 2010. Age was categorised as < 50, 50 to 64, 65 to 79 and ≥ 80 years. Time-trends in the use of TSA were compared using logistic regression or the Cochran Armitage test. The overall use of TSA increased from 2.96/100 000 in 1998 to 12.68/100,000 in 2010. Significantly lower rates were noted between 2009 and 2010, compared with between 1998 and 2000, for: mortality, 0.1% versus 0.2% (p = 0.004); discharge to an inpatient facility, 13.3% versus 14.5% (p = 0.039), and hospital stay > median, 29.4% versus 51.2% (p < 0.001). The rates of use of TSA/100,000 by age groups, < 50, 50 to 64, 65 to 79 and ≥ 80 years were: 0.32, 4.62, 17.82 and 12.56, respectively in 1998 (p < 0.001); and 0.65, 17.49, 75.27 and 49.05, respectively in 2010 (p < 0.001) with an increasing age-related difference over time (p < 0.001). Across the age categories, there were significant differences in the proportion: discharged to an inpatient facility, 3.2% versus 4.2% versus 14.7% versus 36.5%, respectively in 1998 (p < 0.001) and 1.8% versus 4.3% versus 12.5% versus 35.5%, respectively in 2010 (p < 0.001) and the proportion with hospital stay > median, 39.7% versus 40.2% versus 53% versus 69%, respectively in 1998 (p < 0.001) and 17.2% versus 20.6% versus 28.7% versus 50.7%, respectively in 2010 (p < 0.001). In a nationally representative sample, we noted a time-related increase in the use of TSA and increasing age-related differences in outcomes indicating a changing epidemiology of the use of TSA. Age-related differences in outcomes suggest that attention should focus on groups with the worst outcomes.


[Effects of overexpression of miR-17-92 gene cluster on the biological characteristics of prostate cancer cells and its mechanism].
Objective: To explore the effect and mechanism of over-expression of miR-17-92 gene cluster on the biological characteristics of prostate cancer cells. Methods: DU145 cells were transfected with miR-17-92 gene expression plasmid and clones with stable ectopic miR-17-92 overexpression were established. The cell viabilities of DU145-17-92 and DU145-control cells were monitored by xCELLigence system. Cell proliferation and apoptosis were analyzed by Ki-67 and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Cell cycle was detected by flow cytometry. Expression levels of proteins involved in apoptosis and Akt pathway were determined by western blotting. Results: xCELLigence RTCA array data showed that the growth rate of DU145-17-92 cells was significantly higher than that of DU145-control cells after 24 h of seeding (P<0.01). The Ki-67-positive rates of the DU145-control group at 24, 48 and 72 hours were (56.57±1.68)%, (85.48±0.26)% and (90.85±2.08)%, respectively. While the Ki-67 positive rates of the DU145-17-92 group at the desired time points were (73.64±0.68)%, (93.43±1.23)% and (97.36±0.86)%, respectively, with a statistically significant difference at 24 hours (P<0.01). The percentages of apoptotic cells of the DU145-control group at 24, 48 and 72 hours were (6.76±0.09)%, (14.51±0.86)% and (20.73±1.64)%, respectively, while the apoptotic percentages of the DU145-17-92 group were (1.86±0.15)%, (7.90±0.40)% and (4.92±0.48)%, respectively. The percentages of apoptotic cells of the DU145-control group at different time were significantly higher than those of DU145-17-92 group (P<0.01 for all). The result of western blotting showed that the protein expression levels of Bcl-2 interacting mediator of cell death (BIM) and phosphatase and tensin homolog deleted on chromosome ten (PTEN) in DU145-control cells were 0.83±0.00 and 0.91±0.00, respectively, significantly higher than 0.16±0.00 and 0.13±0.00 of DU145-17-92 cells (both P<0.01). Overexpression of miR17-92 induced the phosphorylation of protein kinase B (Akt) at Ser473 while no appreciable effect on the phosphorylation of Akt at Thr308. The phosphorylated level of extracellular regulated protein kinases (ERK) in DU145-control cells was 0.21±0.01, significantly lower than 0.72±0.01 of DU145-17-92 cells (P<0.01). Conclusions: Overexpression of miR-17-92 gene plays a pivotal role in growth, proliferation, apoptosis and cell cycle of DU145 cells through down-regulation of apoptotic protein BIM and tumor suppressor PTEN and activation of Akt and ERK signaling pathway.


Developing and psychometric testing of the anaesthesia nursing competence scale.
The competence of nurses in anaesthesia care is important for the quality of anaesthesia nursing care and patient safety. However, there is a lack of psychometrically tested instruments to measure the competence. Therefore, this study aimed to develop and test the psychometric properties of an anaesthesia nursing competence scale (AnestComp) assessing nurses' competence in anaesthesia care. The scale development and psychometric testing had three phases: (1) based on literature reviews and the description of experts, competence areas were identified and items were created; (2) the content validity of the scale was tested by a content expert group, and the scale was pilot tested; and (3) psychometric testing of scale was tested by anaesthesia nurses' (n = 222) and nursing students' (n = 205) self-assessments. The psychometric testing assessed the reliability when using Cronbach's α and the construct validity using factor analyses (confirmatory and exploratory) and known-group technique. Nursing students were included for the purpose of construct validity testing. The AnestComp has 39 items and consists of seven competence areas: (a) ethics of anaesthesia care, (b) patient's risk care, (c) patient engagement with technology, (d) collaboration within patient care, (e) anaesthesia patient care with medication, (f) peri-anaesthesia nursing intervention, and (g) knowledge of anaesthesia patient care. Cronbach's α values were high in all categories (0.83-0.95), and factor analyses and known-group technique supported a seven-factor model. The initial results supported the reliability and construct validity of the AnestComp. The scale is considered a promising instrument for measuring anaesthesia nursing competence among anaesthesia nurses. Further research with larger and more diverse samples is suggested to refine the current psychometric evaluation.


Hydrophobic forces and the length limit of foldable protein domains.
To find the native conformation (fold), proteins sample a subspace that is typically hundreds of orders of magnitude smaller than their full conformational space. Whether such fast folding is intrinsic or the result of natural selection, and what is the longest foldable protein, are open questions. Here, we derive the average conformational degeneracy of a lattice polypeptide chain in water and quantitatively show that the constraints associated with hydrophobic forces are themselves sufficient to reduce the effective conformational space to a size compatible with the folding of proteins up to approximately 200 amino acids long within a biologically reasonable amount of time. This size range is in general agreement with the experimental protein domain length distribution obtained from approximately 1,200 proteins. Molecular dynamics simulations of the Trp-cage protein confirm this picture on the free energy landscape. Our analytical and computational results are consistent with a model in which the length and time scales of protein folding, as well as the modular nature of large proteins, are dictated primarily by inherent physical forces, whereas natural selection determines the native state.


Adaptive digital notch filter design on the unit circle for the removal of powerline noise from biomedical signals.
This paper investigates adaptive digital notch filters for the elimination of powerline noise from biomedical signals. Since the distribution of the frequency variation of the powerline noise may or may not be centered at 60 Hz, three different adaptive digital notch filters are considered. For the first case, an adaptive FIR second-order digital notch filter is designed to track the center frequency variation. For the second case, the zeroes of an adaptive IIR second-order digital notch filter are fixed on the unit circle and the poles are adapted to find an optimum bandwidth to eliminate the noise to a pre-defined attenuation level. In the third case, both the poles and zeroes of the adaptive IIR second-order filter are adapted to track the center frequency variation within an optimum bandwidth. The adaptive process is considerably simplified by designing the notch filters by pole-zero placement on the unit circle using some suggested rules. A constrained least mean-squared (CLMS) algorithm is used for the adaptive process. To evaluate their performance, the three adaptive notch filters are applied to a powerline noise sample and to a noisy EEG as an illustration of a biomedical signal.


[Inhalation and intravenous anesthesia in pediatric patients].
Inhalation as well as total intravenous anesthesia have both advantages and disadvantages. The pros of an inhalation technique are mask induction without the initial need for intravenous access and precise dosing; the pros of an intravenous technique are postoperative quiet recovery and a low incidence of vomiting. With both techniques the aim is to reach a certain, most often constant effect site concentration, which after a short latent period equals the blood concentration. Initially, with both techniques a higher dosing is required to wash in the compound. An inhalational technique allows the end-tidal concentration to be measured and dosing to be adjusted. Dosing of intravenous anesthetics is based on assumptions. For neonates and young infants an inhalation technique is often preferred because of metabolic immaturity and the resulting difficulties of dosing, whereas older children can often profit from a quiet awakening and a reduced incidence of vomiting. The increased availability of syringe pumps with incorporated algorithms as well as of electroencephalograph (EEG)-based monitoring systems will further promote the popularity of total intravenous anesthesia.


A selective inhibitor of cell proliferation from normal serum.
A factor in normal serum that selectively and reversibly inhibits proliferation of cells in culture has been enriched 160-fold from calf serum by sequential ammonium sulfate precipitation, gel filtration, and lectin-affinity chromatography. DNA synthesis of normal (but not transformed) rat hepatocytes, human lymphoblast lines, and mitogen-stimulated murine spleen cells is inhibited by greater than 90%, and Vero, murine myeloma, MELC, and a human colon carcinoma cell line to a lesser extent. Growth of other cell lines tested was not affected. Responsive cells are arrested apparently in G1 by this inhibitor, the effect of which is maximal by 24 hr and is spontaneously reversible thereafter unless it is renewed. The active fraction is a protein that migrates with the alpha 2-globulins; it is not a lipoprotein, and it is of high apparent molecular weight.


Identification of cell subpopulations by dual-color surface immunofluorescence using biotinylated and unlabeled monoclonal antibodies.
A new method of dual-color immunofluorescence is presented for analysis of surface antigen distribution among heterogeneous cell suspensions. It involves flow cytometric analysis of cells stained with a biotinylated first monoclonal antibody and/or with an unlabeled second monoclonal antibody. After addition of streptavidin-phycoerythrin and/or fluoresceinated goat antimouse immunoglobulin antibody, single-cell fluorescence intensities are measured and biparametric graphic representations are obtained, allowing one to determine the percentage of cells stained by each of the monoclonal antibodies or both. The validity of the method was assessed on human peripheral blood mononuclear cells by using three sets of two monoclonal antibodies: CD8 and CD5, CD3 and CD4, CD11 and HLA-DR. The results showed that dual staining did not induce significant quenching or competition between pairs of antibodies. The procedure is simple and sensitive. It requires only minute amounts of monoclonal antibodies. It is readily applicable to the screening of hybridoma supernatants and to the characterization of new antibodies to cell surface antigens with respect to well-defined markers.


Predictors of prolonged benefit from palbociclib plus fulvestrant in women with endocrine-resistant hormone receptor-positive/human epidermal growth factor receptor 2-negative metastatic breast cancer in PALOMA-3.
The addition of palbociclib to fulvestrant improved clinical outcomes over placebo-fulvestrant in endocrine-pretreated metastatic breast cancer (MBC) patients in PALOMA-3. Here, we examined factors predictive of long-term benefit. Premenopausal-peri/postmenopausal patients with endocrine-resistant, hormone receptor-positive (HR+)/human epidermal growth factor receptor 2-negative MBC were randomised 2:1 to fulvestrant (500 mg) and either palbociclib (125 mg/d; 3/1 schedule; n = 347) or placebo (n = 174). Baseline characteristics, mutation status and HR expression levels were compared in patients with and without prolonged benefit (treatment duration ≥18 months). By August 2016, 100 patients (29%) on palbociclib-fulvestrant and 26 (15%) on placebo-fulvestrant demonstrated prolonged benefit, with long-term responders in both arms sharing common clinical characteristics. They usually had less disease burden at baseline versus those treated <18 months, such as having one disease site (40% vs 29% on palbociclib-fulvestrant and 69% vs 29% on placebo-fulvestrant), bone-only disease (32% vs 22% and 46% vs 17%) and were less heavily pretreated (69% vs 56% and 73% vs 60% had ≤2 prior therapies). Baseline tumour ESR1 and PIK3CA mutation rates were lower among long-term responders in both arms; median oestrogen receptor H-scores were similar, whereas progesterone receptor H-scores were higher among long-term responders. This exploratory analysis demonstrates that some patients with endocrine-resistant MBC derive significant and prolonged benefit when treated with palbociclib-fulvestrant, with fewer patients experiencing similar efficacy with placebo-fulvestrant. The current analysis did not identify specific molecular or clinical factors prognostic of long-term benefit with palbociclib-fulvestrant (ClinicalTrials.gov, NCT01942135).


Non-thermal emerging technologies as alternatives to chemical additives to improve the quality of wheat flour for breadmaking: a review.
Wheat flour is the main ingredient used in the preparation of bread. Factors such as low gluten content and the addition of nontraditional ingredients in baking affect the quality of wheat flour and may limit its use in baking. With the increasing trend of "clean label" products, it may be interesting to develop and use physical processes to improve the quality of wheat flour and avoid the use of chemical additives. High hydrostatic pressure, non-thermal plasma, ultrasound, ozonation, ultraviolet light, and pulsed light treatments are non-thermal emerging technologies (NTETs) that have been studied for this purpose. They were originally developed to inactivate microorganisms and enzymes in foods. Additionally, these technologies can be used at low temperatures to modify the most important component of wheat flour, i.e., gluten and its fractions, which are responsible for the rheological properties of wheat flour dough. Thus, this review focuses on the effects of these NTETs by considering the following factors: (1) the technological properties of gluten, (2) gluten-starch interactions, (3) possible effects of NTETs on minor components of flours, and (4) the quality of wheat flour and the resulting final products.


Characteristics of biochemical markers in patients with metabolic bone disorders.
Biochemical markers of bone turnover are expected to have some different characteristics among bone metabolic disorders. We compared bone formation markers: serum total alkaline phosphatase (s-Alp), serum osteocalcin (s-OC) and serum carboxy-terminal propeptide of type I collagen (s-PICP); and bone resorption markers: serum carboxy-terminal telopeptide of type I collagen (s-ICTP), urinary pyridinoline (u-Pyr) and urinary deoxypyridinoline (u-Dpyr) to examine which marker is the most suitable and reliable to evaluate bone turnover in patients with osteoporosis (n = 29), osteomalacia (n = 10), primary hyperparathyroidism (n = 6) and renal osteodystrophy (n = 21). The value of s-Alp in the osteomalacia group was significantly higher than those in the normal control group and the osteoporosis group (p < 0.001), and T-score of s-Alp was significantly higher than those of s-OC and s-PICP in the osteomalacia group. The values of u-Pyr and u-Dpyr in the primary hyperparathyroidism group were significantly higher than those in the other groups (p < 0.001). S-PICP, which are not dependent upon renal function, was much higher in the renal osteodystrophy group than in all other groups. In the osteoporosis group, T-score of s-ICTP was significantly higher than those of s-OC. Thus, s-Alp was a good marker in osteomalacia, u-Pyr and u-Dpyr in primary hyperparathyroidism, s-PICP in renal osteodystrophy, and s-ICTP in osteoporosis.


Second messenger function and the structure-activity relationship of cyclic adenosine diphosphoribose (cADPR).
Cyclic ADP-ribose (cADPR) is a Ca2+ mobilizing second messenger found in various cell types, tissues and organisms. Receptor-mediated formation of cADPR may proceed via transmembrane shuttling of the substrate NAD and involvement of the ectoenzyme CD38, or via so far unidentified ADP-ribosyl cyclases located within the cytosol or in internal membranes. cADPR activates intracellular Ca2+ release via type 2 and 3 ryanodine receptors. The exact molecular mechanism, however, remains to be elucidated. Possibilities are the direct binding of cADPR to the ryanodine receptor or binding via a separate cADPR binding protein. In addition to Ca2+ release, cADPR also evokes Ca2+ entry. The underlying mechanism(s) may comprise activation of capacitative Ca2+ entry and/or activation of the cation channel TRPM2 in conjunction with adenosine diphosphoribose. The development of novel cADPR analogues revealed new insights into the structure-activity relationship. Substitution of either the northern ribose or both the northern and southern ribose resulted in much simpler molecules, which still retained significant biological activity.


The skin test antigen stimulated killer (STAK) cell mediating NK like CMC is OKM1 positive and OKT3 negative.
Recently we demonstrated that candida antigen stimulated natural killer cell like cell-mediated cytolysis (NK like CMC) in peripheral blood mononuclear cells (PBMNC) isolated from normal individuals (Tartof et al., 1980). Utilizing monoclonal antibodies directed against human mononuclear cell subpopulations in conjunction with a fluorescence activated cell sorter (FACS) we determined that, similar to the previously described NK cell, the skin test antigen stimulated killer (STAK) cell is a larger OKM1 positive, OKT3 negative cell. We obtained similar results using two different skin test antigens. Thus, stimulation of NK like CMC in PBMNC by skin test antigens probably represents activation of NK or NK like cells.


Contribution of clonal dissemination and selection of mutants during therapy to Pseudomonas aeruginosa antimicrobial resistance in an intensive care unit setting.
Rates of antibiotic resistance in Pseudomonas aeruginosa isolates from intensive care unit (ICU) patients are expected to be dependent on the selection of resistance mutations during therapy, the availability of exogenous resistance determinants and their dissemination potential, and the efficiency of transmission of the resistant strains. The relative contributions of these three factors were studied in an ICU with no apparent outbreak in 216 sequential P. aeruginosa isolates recovered from 102 patients between September 2002 and November 2003. Analysis of pulsed-field gel electrophoresis patterns revealed the presence of 82 different clones. Thus, the dissemination of particular resistant clones had a minimal effect on the relatively high overall resistance frequencies found for imipenem (32%), cefepime (25%), ceftazidime (24%), meropenem (22%), ciprofloxacin (18%) and tobramycin (2%). Rates of primary resistance were relatively low, and resistance development during treatment (secondary resistance) was the main factor contributing to the overall high resistance rates. In ICU settings with a low prevalence of epidemic resistant strains, the main strategy for resistance control should focus on the design of targeted regimens to avoid the development of resistance.


Using Preanesthesia Clinic Visits to Improve Advance Directives Completion: An Interrupted Time Series Analysis.
Advance directives documentation can increase the likelihood that patient's wishes are respected if they become incapacitated. Unfortunately, completion rates are suboptimal overall, and disparities may exist, especially for vulnerable groups. We assessed whether implementing an initiative to standardize advance directives discussions during preanesthesia visits was associated with changes in rates of advance directives completion over time, and whether the association depends on race, insurance type, or income. We conducted a before-after interrupted time series evaluation between January 1, 2015 and June 30, 2019 in a single-center, outpatient preanesthesia clinic. Participants were adults who visited the preanesthesia clinic at Cleveland Clinic and had >1 comorbidity before a noncardiac surgery of either medium or high risk. The intervention in March of 2017 consisted of training staff to help patients complete and witness advance directives documents during visits. We measured advance directives completion, by race, payor, and income (using the 2019 Federal Poverty Line). We assessed the confounder-adjusted association between intervention (pre versus post) and proportion of patients completing advanced directives over time using segmented regression to compare slopes between periods and assess changes at start of the intervention. We used similar models to assess whether changes depended on race, insurance type, or income level. We included 26,368 visits from 22,430 patients. We analyzed financial status for 16,788 visits from 14,274 patients who had address data. There were 11,242 (43%) visits preintervention and 15,126 (57%) visits postintervention. Crude completion rates for advance directives increased from 29% to 78%, with odds of completion an estimated 18 times higher than preintervention (odds ratio [95% CI] of 18 [16-21]; P < 0.001). Regarding race, Black patients had lower completion rates preintervention than White patients, although the gap steadily closed after the intervention ( P = .001). Postintervention, both race groups immediately increased, with no difference in amount of increase ( P = .17) or postintervention change in slope difference ( P = .17). Regarding insurance, patients with Medicaid had lower preintervention completion rates than those with private. Intervention was associated with increases in both groups, but the difference in slopes ( P = .43) or proportions ( P = .23) between the groups did not change after intervention. Regarding the Federal Poverty Line, the completion rate gap between those below (<100%) and above (139%-400%) narrowed by approximately half (0.51: 95% CI, 0.27-0.98; P = .04). Standardizing advance directives discussions during preanesthesia visits was associated with more patients completing advance directives, particularly in vulnerable patient groups.


Oral antiseptic and periodontitis: a clinical and microbiological study.
This study evaluated the efficacy of a xanthan-based chlorhexidine gel (Xan-CHX) used as antiseptic in periodontal pockets after scaling and root planing in patients with generalized mild-severe chronic periodontitis. 30 systemically healthy patients, 15 males and 15 females (mean age 54,1 ± 6,9 years), diagnosed for chronic periodontitis were enrolled in this study. For each patient, in the 1(st) and 4(th) quadrant was applied antiseptic (Test sites); while the 2nd and 3rd were treated with the only mechanical causal therapy without using chemical agents (Control sites). Sub gingival plaque, gingival bleeding (BOP+), plaque index (PI-Plaque Control Record) and Probing Depths (PD) were evaluated at baseline (prior to any treatment) and after 4 weeks (21 days after treatment). The results showed that the mechanical causal therapy has a good effect in reducing the clinical indices, but the addition of antiseptics provides a significantly improvement in the PI, BOP and frequency of periodontopathic bacteria. The application of xanthan-based chlorhexidine (Xan-CHX) gel offers a great benefit in improving of the indices of periodontal disease, proving to be essential as adjunctive therapy in patients with a serious or moderate high chronic periodontitis, and of course as part of a program of periodontal treatment.


Muscle blood flow in humans: how high can it go?
The question is whether skeletal muscle blood flow (MBF) in humans can reach the peak values of 300-400 ml.100 g-1.min-1 seen in dogs, ponies, and rats. The answer depends on the total mass of active muscle. Total MBF has a fixed upper limit or maximal value set by maximal cardiac output--normally about 25 1.min-1. In a 75-kg man with 30 kg of muscle, total MBF is 22 1.min-1 or 73 ml.100 g-1.min-1. As the mass of active muscle decreases, MBF available per 100 g of muscle increases, but a "maximal" value can no longer be defined because it becomes a function of blood pressure and muscle vascular conductance (and not maximal cardiac output) for which "maximal" values are also undefined. This paper emphasizes three key points. First, pumping capacity of the human heart is much smaller per kg body weight than in many other species. Second, human cardiac pumping capacity is exceeded by the vasodilator capacity of skeletal muscle; thus, so-called "peak MBF" often occurs when metabolic vasodilator and blood pressure-regulating vasoconstrictor mechanisms are competing. Third, in order to observe a peak MBF, one must maximally activate a mass of muscle that is too small to overwhelm cardiac pumping capacity. This experimental approach has revealed that MBF can exceed 300 ml.100 g-1.min-1 in small masses of active human muscle. The upper limit for flow, which depends on pressure, and for vascular conductance is still unknown.


Toward the comparison of rare earth element and actinide behavior in materials: a computational study of Ce- and U-bearing britholites.
We present a computational investigation into the nature of bonds formed by f-elements in materials. The paper presents an example of the incorporation of rare earth elements (REE) and actinides in minerals derived from fluorapatite: Ca(10)(PO(4))(6)F(2). These minerals, called britholites, allow many substitutions on all three Ca, P, and F sites and are considered as potential host phases for radioactive elements separated from nuclear waste. REE and actinides have very similar physical and chemical properties, but REE are not radioactive and much more easily handled. REE are, therefore, very often used as a surrogate for actinides in experimental studies. The representative elements of rare earths and actinides chosen for this first investigation are cerium and uranium, respectively. We have studied all the various configurations of Ca(9)X(PO(4))(6)(-)(y)()(SiO(4))(y)()F(2), where X stands for Ce(3+), Ce(4+), U(3+), and U(4+), and y is equal to 1 and 2 for three-time and four-time charged cations, respectively. Calculations have been performed within the density functional theory (DFT) framework according to the computation scheme determined in a previous study. The analysis of the energies of the various configurations shows that the incorporation of all the cations considered stabilizes the apatitic structure. This stabilization, however, is greater for four-time charged cations than for three-time charged ones, which shows that Ce and U are both preferentially substituted in the +IV oxidation state. In addition, the substitution in one of the two cationic sites of the apatitic structure is always more favorable. Then, the geometry analysis shows a larger decrease in size of this cationic site for U than for Ce, as well as different volume variations for Ce and U substitutions in the two cationic sites. This cannot be explained by steric effects alone. Finally, the electronic density analysis yields three essential results: U and Ce form significantly covalent bonds, U forms bonds more covalent than Ce, and finally four-time charged cations form more covalent bonds than three-time charged ones. The comparison of these results with the formation enthalpies of the various phases shows a positive correlation between the covalence degree of the bonds formed by the f-element and the stability of the structure. In addition, our results prove that Ce- and U-bearing britholites exhibit very similar energetic, structural, and electronic properties. Ce, therefore, appears to be a good simulant for U.


Massive blood transfusion and outcome in 1062 polytrauma patients: a prospective study based on the Trauma Registry of the German Trauma Society.
About 15% of polytrauma patients receive massive blood transfusion (MBT) defined as > or = 10 units of packed red blood cells (PRBC). In general, the prognosis of trauma patients receiving MBT is considered to be poor. The purpose of this study was to investigate the impact of MBT on the outcome of polytrauma patients. Records of 10 997 patients in the Trauma Registry of the German Trauma Society were analysed. Transfusion data were available from 8182 severe trauma patients with a mean injury severity score of 24.5 and, of these 8182 patients, 1062 received > or = 10 units of PRBC. First, a logistic regression model for the predictors of mortality was performed. Second, incidences of organ failure and sepsis as well as survival rates were analysed. The highest risk for mortality was age over 55 years (odds ratios [OR] 4.7; confidence intervals [CI 95%], 3.5-6.5) followed by Glasgow Coma Scale < or = 8 (OR 4.6; 3.4-6.1), MBT > or = 20 units of PRBC (OR 3.3; 2.1-5.4), thromboplastin time < 50% (OR 3.2; 2.2-4.4) and injury severity score > or = 24 (OR 2.9; 2.1-4.1). Transfusion of 10-19 PRBC was identified as the variable with the lowest risk for mortality (OR 1.5; 1.0-2.3). Risk of organ failure, sepsis and death correlated with increasing transfusion amount. For the MBT patients, the survival rate was 56.9% (CI 95%, 53.9-59.9%) compared to 85.2% (84.4-86.0%) of non-MBT patients (P < 0.001). In the MBT group with > 30 PRBC (mean 40.6 PRBC) 39.6% survived (31.7-47.5%). Massive blood transfusion is one main prognostic factor for mortality in trauma. Although MBT is generally considered to be critical, every second trauma patient with MBT survived. A cut-off value for the number of PRBC could not be determined. Extended transfusion management even with high amounts of PRBC seems to be justified.


Auditory cortical responses in hearing subjects and unilateral deaf patients as detected by functional magnetic resonance imaging.
Functional magnetic resonance imaging is a non-invasive method for the detection of focal brain activity at high spatial resolution. Acoustic stimulation leads to a blood oxygenation level dependent signal change in the plane of the superior temporal gyrus. The dependence of this response in the auditory cortex on binaural, monaural left and monaural right acoustic stimulation for 10 healthy subjects and five monaural deaf patients is described. Acoustic stimulation consists of 1000 Hz pulsed sine tones at a pulse rate of 6 Hz and a sound pressure level of 95 dB. For monaural stimulation, normal-hearing subjects revealed a strong lateralization of cortical response towards the contralateral hemisphere. The lateralization ratios between left and right hemispheric response areas were 3.4-5.2 for monaural stimulation and nearly balanced for binaural stimulation. Additionally, the sum of cortical activation volumes induced by monaural left and right stimulation was approximately 30% smaller than for binaural stimulation, indicating either inhibitory mechanisms or neuronal facilitation within the auditory pathways. For monaural deaf subjects the lateralization ratio between left to right response was just 1.3 towards the contralateral hemisphere of the healthy ear, which is comparable to binaural responses of normal-hearing subjects. This observation seems to indicate a plasticity or a reorganization of auditory pathways of monaural deaf patients.


Comparative genome analysis of Bacillus sphaericus by ribotyping, M13 hybridization, and M13 polymerase chain reaction fingerprinting.
A comparative genome analysis of 15 strains representing the five homology groups of the highly heterogeneous species Bacillus sphaericus was performed by M13 hybridization fingerprinting, M13 polymerase chain reaction fingerprinting, and ribotyping with the whole rrn operon. The computer cluster analyses of the polymorphic patterns, presented in dendrograms, showed that these methods allow the differentiation of the individual strains and some homology groups. Our results confirm the close genetic relatedness of the mosquito pathogenic strains of group IIA and support the idea for differentiation of a separate species. At the same time, we present additional proof of the significant genetic heterogeneity of B. sphaericus and the necessity of reconsideration of its present classification.


Scattering coefficient determination in turbid media with backscattered polarized light.
A simple empirical method is presented to determine the scattering coefficient mu' s from backscattered polarized images of turbid media. It uses the ratio, pixel by pixel, of two images that are the second and the first backscattered Stokes parameter images Q and I, respectively. Taking this image ratio, then integrating it over the azimuth angle, we get a function depending on the distance from the light entrance point. This function has a maximum. Using Monte Carlo simulations, for a fixed reduced scattering coefficient mu s and for an anisotropy factor g varying between 0 and 0.8, it is found a linear relationship between the scattering coefficient mu s and the inverse of the maximum position of this function.


Traumatic unilateral jumped facet joint in the upper thoracic spine: Case presentation and literature review.
A jumped facet joint is defined by when the inferior articular process of the superior vertebra becomes locked anterior to the superior articular process of the inferior vertebra. These typically traumatic lesions are exceedingly rare in the thoracic spine. Here, we present a patient with a unilateral jumped facet joint in the upper thoracic spine treated with open reduction and an instrumented fusion. A 45-year-old male presented after a significant motor vehicle accident. In the emergency room, he had a Glasgow Coma Score of 13 without any neurologic deficit. The thoracic computed tomography (CT) showed a significant jumped left facet at the T2-T3 level. Two days later, utilizing intraoperative CT-guided navigation and neuromonitoring, he underwent open reduction of the T2-T3 jumped facet plus an instrumented T1-T5 fusion. X-rays taken 3-month postoperatively showed a stable construct. Six months postoperatively, he remained neurologically intact. A unilateral jumped thoracic facet may be present in patients with fractured ribs. The mechanism of injury is most likely axial rotation. Both CT and magnetic resonance imaging studies allow for early detection of these very rare lesions and warrant open reduction and instrumented fusion.


Charge-Order-Induced Ferroelectricity in LaVO_{3}/SrVO_{3} Superlattices.
The structure and properties of the 1∶1 superlattice of LaVO_{3} and SrVO_{3} are investigated with a first-principles density-functional-theory-plus-U (DFT+U) method. The lowest energy states are antiferromagnetic charge-ordered Mott-insulating phases. In one of these insulating phases, layered charge ordering combines with the layered La/Sr cation ordering to produce a polar structure with a large nonzero spontaneous polarization normal to the interfaces. This polarization, comparable to that of conventional ferroelectrics, is produced by electron transfer between the V^{3+} and V^{4+} layers. The energy of this normal-polarization state relative to the ground state is only 3 meV per vanadium. Under tensile strain, this energy difference can be further reduced, suggesting that the normal-polarization state can be induced by an electric field applied normal to the superlattice layers, yielding an antiferroelectric double-hysteresis loop. If the system does not switch back to the ground state on removal of the field, a ferroelectric-type hysteresis loop could be observed.


Genetic assessment of leech species from yak (Bos grunniens) in the tract of Northeast India.
Yak is an iconic symbol of Tibet and high altitudes of Northeast India. It is highly cherished for milk, meat, and skin. However, yaks suffer drastic change in milk production, weight loss, etc, when infested by parasites. Among them, infestation by leeches is a serious problem in the Himalayan belt of Northeast India. The parasite feeds on blood externally or from body orifices, like nasopharynx, oral, rectum, etc. But there has been limited data about the leech species infesting the yak in that region because of the difficulties in morphological identification due to plasticity of the body, changes in shape, and surface structure and thus, warrants for the molecular characterization of leech. In anticipation, this study would be influential in proper identification of leech species infesting yak track and also helpful in inventorying of leech species in Northeast India. Here, we investigated, through combined approach of molecular markers and morphological parameters for the identification of leech species infesting yak. The DNA sequences of COI barcode fragment, 18S and 28S rDNA, were analyzed for species identification. The generated sequences were subjected to similarity match in global database and analyzed further through Neighbour-Joining, K2P distance based as well as ML approach. Among the three markers, only COI was successful in delineating species whereas the 18S and 28S failed to delineate the species. Our study confirmed the presence of the species from genus Hirudinaria, Haemadipsa, Whitmania, and one species Myxobdella annandalae, which has not been previously reported from this region.


ISH and PCR study with Y-specific probe/primers.
Our main aim was to evaluate whether maternal whole venous blood could be used for determination of fetal sex, when no enrichment of fetal cells was attempted and when "standard" interphase cytogenetics and PCR analysis were adopted. Altogether 39 pregnant women were studied by using ISH and 59 by using PCR. Out of the 59 pregnant women, 26 carried a male fetus and 33 a female fetus. By ISH, Y-positive cells were detected in 12 of 19 pregnancies with a male fetus and in two of the 20 pregnancies with a female fetus. The frequency of the fetal cells ranged from 1 in 639 to 1 in 100,000. By nested PCR with primers flanking a Y-specific repeat sequence, the positive band indicating a male fetus was found in one of the 26 pregnancies with a male fetus and in one of the 33 pregnancies with a female fetus. According to our results, fetal cells in maternal blood cannot be reliably used for prenatal diagnosis without enrichment of fetal cells.


Urine methanol concentration and alcohol hangover severity.
Congeners are substances, other than ethanol, that are produced during fermentation. Previous research found that the consumption of congener-rich drinks contributes to the severity of alcohol hangover. Methanol is such a congener that has been related to alcohol hangover. Therefore, the aim of this study was to examine the relationship between urine methanol concentration and alcohol hangover severity. N = 36 healthy social drinkers (22 females, 14 males), aged 18-30 years old, participated in a naturalistic study, comprising a hangover day and a control day (no alcohol consumed the previous day). N = 18 of them had regular hangovers (the hangover group), while the other N = 18 claimed to be hangover-immune (hangover-immune group). Overall hangover severity was assessed, and that of 23 individual hangover symptoms. Urine methanol concentrations on the hangover and control days were compared, and correlated to hangover (symptom) severity. Urine methanol concentration was significantly higher on hangover days compared to control days (p = 0.0001). No significant differences in urine methanol concentration were found between the hangover group and hangover-immune group. However, urine methanol concentration did not significantly correlate with overall hangover severity (r = -0.011, p = 0.948), nor with any of the individual hangover symptoms. These findings were observed also when analyzing the data separately for the hangover-immune group. In the hangover group, a significant correlation with urine methanol concentration was found only with vomiting (r = 0.489, p = 0.037). No significant correlation was observed between urine methanol concentration and hangover severity, nor with individual core hangover symptoms.


The intercalated nuclear complex of the primate amygdala.
The organization of the inhibitory intercalated cell masses (IM) of the primate amygdala is largely unknown despite their key role in emotional processes. We studied the structural, topographic, neurochemical and intrinsic connectional features of IM neurons in the rhesus monkey brain. We found that the intercalated neurons are not confined to discrete cell clusters, but form a neuronal net that is interposed between the basal nuclei and extends to the dorsally located anterior, central, and medial nuclei of the amygdala. Unlike the IM in rodents, which are prominent in the anterior half of the amygdala, the primate inhibitory net stretched throughout the antero-posterior axis of the amygdala, and was most prominent in the central and posterior extent of the amygdala. There were two morphologic types of intercalated neurons: spiny and aspiny. Spiny neurons were the most abundant; their somata were small or medium size, round or elongated, and their dendritic trees were round or bipolar, depending on location. The aspiny neurons were on average slightly larger and had varicose dendrites with no spines. There were three non-overlapping neurochemical populations of IM neurons, in descending order of abundance: (1) Spiny neurons that were positive for the striatal associated dopamine- and cAMP-regulated phosphoprotein (DARPP-32+); (2) Aspiny neurons that expressed the calcium-binding protein calbindin (CB+); and (3) Aspiny neurons that expressed nitric oxide synthase (NOS+). The unique combinations of structural and neurochemical features of the three classes of IM neurons suggest different physiological properties and function. The three types of IM neurons were intermingled and likely interconnected in distinct ways, and were innervated by intrinsic neurons within the amygdala, or by external sources, in pathways that underlie fear conditioning and anxiety.


Complete Genome Sequence of Streptococcus pneumoniae Virulent Phage MS1.
The lytic Streptococcus pneumoniae phage MS1 was isolated from a throat swab of a patient with symptoms of upper respiratory tract infection. The genome of this siphophage has 56,075 bp, 42.3% G+C content, and 77 open reading frames, including queuosine biosynthesis genes. Phage MS1 is related to pneumococcal phage Dp-1.


Women's lived experience of infertility after unsuccessful medical intervention.
The purpose of this qualitative descriptive study was to explore the phenomenon of women's experience with infertility in the aftermath of unsuccessful medical treatment. A purposive sample of 22 women between the ages of 33 and 48 years participated in a hermeneutic-phenomenological research process and were interviewed an average of 3.9 years after unsuccessful medical treatment. Women described the existential challenges to their sense of self, their identity, and the meaning and purpose of life. The paradoxical dimensions of loss and opportunity in their experience contributed to an altered view of themselves and their world. Infertility and its role as a life-defining experience pervaded their stories of living with infertility after unsuccessful treatment. Health care professionals are advised to assess women's overall well-being, mental health status with particular attention to spiritual well-being, and their social support network as a basis for determining supportive services that may be required in the wake of unsuccessful treatment for infertility.


Huntington's disease: diagnosis and management.
Huntington's disease (HD) is an inherited neurodegenerative disease characterised by neuropsychiatric symptoms, a movement disorder (most commonly choreiform) and progressive cognitive impairment. The diagnosis is usually confirmed through identification of an increased CAG repeat length in the huntingtin gene in a patient with clinical features of the condition. Though diagnosis is usually straightforward, unusual presentations can occur, and it can be difficult to know when someone has transitioned from being an asymptomatic carrier into the disease state. This has become increasingly important recently, with several putative disease-modifying therapies entering trials. A growing number of conditions can mimic HD, including rare genetic causes, which must be considered in the event of a negative HD genetic test. Patients are best managed in specialist multidisciplinary clinics, including when considering genetic testing. Current treatments are symptomatic, and largely directed at the chorea and neurobehavioural problems, although supporting trial evidence for these is often limited.


Reduction of influenza virus titer and protection against influenza virus infection in infant mice fed Lactobacillus casei Shirota.
We investigated whether oral administration of Lactobacillus casei strain Shirota to neonatal and infant mice ameliorates influenza virus (IFV) infection in the upper respiratory tract and protects against influenza infection. In a model of upper respiratory IFV infection, the titer of virus in the nasal washings of infant mice administered L. casei Shirota (L. casei Shirota group) was significantly (P < 0.05) lower than that in infant mice administered saline (control group) (10(2.48) +/- 10(0.31) and 10(2.78) +/- 10(0.4), respectively). Further, the survival rate of the L. casei Shirota group was significantly (P < 0.05) higher than that of the control group (14.3 versus 40.0%). One day after infection, pulmonary NK cell activity and interleukin-12 production by mediastinal lymph node cells of mice in the L. casei Shirota group were significantly greater than those of mice in the control group. These findings suggest that oral administration of L. casei Shirota activates the immature immune system of neonatal and infant mice and protects against IFV infection. Therefore, oral administration of L. casei Shirota may accelerate the innate immune response of the respiratory tract and protect against various respiratory infections in neonates, infants, and children, a high risk group for viral and bacterial infections.


The impact of contact tracing in clustered populations.
The tracing of potentially infectious contacts has become an important part of the control strategy for many infectious diseases, from early cases of novel infections to endemic sexually transmitted infections. Here, we make use of mathematical models to consider the case of partner notification for sexually transmitted infection, however these models are sufficiently simple to allow more general conclusions to be drawn. We show that, when contact network structure is considered in addition to contact tracing, standard "mass action" models are generally inadequate. To consider the impact of mutual contacts (specifically clustering) we develop an improvement to existing pairwise network models, which we use to demonstrate that ceteris paribus, clustering improves the efficacy of contact tracing for a large region of parameter space. This result is sometimes reversed, however, for the case of highly effective contact tracing. We also develop stochastic simulations for comparison, using simple re-wiring methods that allow the generation of appropriate comparator networks. In this way we contribute to the general theory of network-based interventions against infectious disease.


Estimating the number of deaths averted from 2008 to 2020 within the Ethiopian CMAM programme.
The management of wasting in Ethiopia is heavily reliant on the Community-based Management of Acute Malnutrition (CMAM) programme that has been implemented in more than 18,000 service delivery points scattered across the country. Despite the full-scale implementation of the CMAM, the number of child death averted, and the cost per child death averted remains unknown. This study aimed to estimate the cost and the number of child death averted by the CMAM programme between 2008 and 2020. Using data from routine monitoring of the CMAM programme, we estimated the excess mortality averted by the programme and estimated the cost per averted child death based on supply and labour. Over the past 13 years between 2008 and 2020, 3.6 million children under 5 years were admitted to the Ethiopian CMAM programme. The yearly average admission of 317,228 was achieved since 2011. On average, ~34,000 child deaths were averted yearly. The CMAM programme was estimated to have saved 437,654 (95% confidence interval [CI]: 320,161; 469,932) child deaths between 2008 and 2020, approximately 12% of the admitted cases. The average cost of the programme per adverted death was estimated at US$762/child death averted (95% CI = 639; 1001). The CMAM programme in Ethiopia is cost-effective and has continued to avert a significant number of child death. Given the high short- and long-term economic and health consequences of child wasting, concerted multi-sectoral efforts are needed to accelerate progress not only in its treatment but also in its prevention.


Association of MUC16 Mutation With Tumor Mutation Load and Outcomes in Patients With Gastric Cancer.
MUC16, which encodes cancer antigen 125 (CA-125), is frequently mutated in gastric cancer (GC); however, its association with tumor mutation load (TML) and outcome in patients with GC has not been established, to date. To investigate whether MUC16 mutations are associated with TML and prognosis in patients with GC. Statistical analysis of genomic data from 437 GC samples obtained from The Cancer Genome Atlas (TCGA) and 256 samples from an Asian cohort. Both cohorts contained data of patients with GC involved in previous genomic studies. Data were obtained from TCGA on September 3, 2017, and from the Asian cohort on March 5, 2013, and analyzed from September 3 to December 1, 2017. The TCGA cohort was used as a discovery set and the Asian cohort as a validation set. Kaplan-Meier survival analysis and multivariate Cox and logistic regression models were applied. Regression models addressed confounding factors; Bayesian variant nonnegative matrix factorization was used to extract mutational signatures. The MutSigCV algorithm was used to identify significantly mutated genes. Primary outcomes were mutation frequency, overall survival, and TML, calculated using Kaplan-Meier survival analysis, odds ratios (ORs), and significance of signaling pathways. MUC16 was mutated in 168 of 437 (38.4%) of the GC samples from the TCGA cohort and in 57 of 256 (22.3%) from the Asian cohort. In both cohorts, GC samples with MUC16 mutations exhibited significantly greater TML than those without MUC16 mutations (median mutation counts: TCGA cohort, 264 with MUC16 mutation vs 115 without; Asian cohort, 134 with MUC16 mutation vs 74 without; Wilcoxon rank sum test, both P < .001). This association was independent of mutations in POLE and BRCA1/2 and mutational signatures in the TCGA cohort (OR, 1.87; 95% CI, 1.49-2.36; P < .001) and the Asian cohort (OR, 1.69; 95% CI, 1.25-2.29; P < .001). MUC16 mutations were significantly associated with better prognosis in both cohorts (median overall survival, 46.9 [95% CI, 26.4-NA (not available)] vs 26.7 [95% CI, 20.2-43.1] months; log-rank test, P = .007 [TCGA cohort] and not calculable [the median overall survival of patients with GC and MUC16 mutations could not be calculated because more than half the patients in the group were alive] vs 36.8 months; P = .04 [Asian cohort]). The association remained statistically significant after controlling for age, sex, TNM stage, mutations in POLE and BRCA1/2, and mutational signatures (hazard ratio, 0.61 [95% CI, 0.42-0.89]; log rank test, P = .01). Immune response and cell cycle regulation circuits were among the top altered signaling pathways in samples with MUC16 mutations (normalized enrichment score, 1.70 [95% CI, 1.57-1.79] and 2.04 [95% CI, 1.90-2.18]; adjusted P < .001). The prognostic significance of MUC16 mutation identified in the TCGA cohort was validated in the Asian cohort. These findings indicate that MUC16 mutations may be associated with higher TML, better survival outcomes, and immune response and cell cycle pathways. These findings may be immediately applicable for guiding immunotherapy treatment for patients with GC.


Which Cholecystectomy do Medical Students Prefer?
This study was undertaken to identify which minimally invasive technique medical students prefer for cholecystectomy and what factors determine their decision. Brazilian medical students watched a video reviewing the advantages and disadvantages of six different surgical approaches to cholecystectomy: open surgery, conventional laparoscopy, mini-laparoscopy (MINI), single-incision laparoscopic surgery, natural-orifice transluminal endoscopic surgery, and robotic surgery. Respondents then answered questions about hypothetical situations in which the participants would be submitted to elective cholecystectomy. One hundred eleven medical students completed the survey, 60 females (54%) and 51 males (46%). Most students were 19-26 years old. When asked whether they would consider an open cholecystectomy if minimally invasive surgery (MIS) techniques were available, only 9% answered yes. Senior medical students were the least willing to consider open surgery (P = .036). When asked if they would prefer conventional laparoscopy, MINI, or robotic surgery for their cholecystectomy, 85% of the women and 63% of the men chose MINI (P = .025). When asked if they would consider a single-incision laparoscopic surgery or natural-orifice transluminal endoscopic surgery approach, 94 respondents (84%) answered no. When asked to rank which factors they consider the most important when choosing a surgical technique, they ranked safety of the procedure first (58%) and surgeon experience second (30%). When Brazilian medical students were asked to select a surgical approach for cholecystectomy, most chose MINI. The preference for MINI was strongest amongst female medical students. Both female and male medical students ranked safety as the most important factor.


[Cryptotanshinone May Induce Ferroptosis of Human Liver Cancer HepG2 Cells].
Objective To observe the effect of cryptotanshinone on the ferroptosis of human liver cancer HepG2 cells. Methods The viability of the HepG2 cells cultured in vitro was determined using the Cell Counting Kit-8(CCK-8),and the half maximal inhibitory concentration(IC50)was calculated.The cell morphology was observed using an inverted microscope.The reactive oxygen species(ROS)level was detected with the 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)probe.The glutathione(GSH)assay kit was used to determine the GSH level.Western blot analysis was employed to detect the expression of cystine/glutamate antiporter system light chain(xCT)and glutathione peroxidase 4(GPX4),two marker proteins in ferroptosis.Additionally,the cell viability,ROS level,GSH level,and the expression levels of xCT and GPX4 were detected for the cells treated with the ferroptosis inhibitor ferrostain-1(Fer-1),the iron chelator deferoxamine(DFO),and the ROS scavenger N-acetylcysteine(NAC).Results Cryptotanshinone significantly inhibited the cell viability of HepG2 cells with an IC50 of 93.73 μmol/L,and caused the morphological changes and death of the cells.It could significantly induce ROS accumulation,reduce GSH level,and down-regulate the expression of xCT and GPX4 in HepG2 cells.Fer-1,DFO,and NAC can remedy the cryptotanshinone-caused decrease in the cell viability of HepG2 cells.Fer-1 could inhibit cryptotanshinone-induced ROS accumulation,restore GSH level,and recover the expression of xCT and GPX4. Conclusion Cryptotanshinone may increase the accumulation of ROS by inhibiting the expression of xCT and GPX4 to induce the ferroptosis of HepG2 cells.


Comparative life cycle assessment study on environmental impact of oil production from micro-algae and terrestrial oilseed crops.
In this study the LCA methodology is applied in order to satisfy two goals: i) to evaluate the hot spots in site-specific production chain of biodiesel from terrestrial and micro-algae feedstock; ii) to compare quantitatively, utilizing primary data, the impacts of the first generation in respect to the third generation bio-fuels. Results show that micro-algae are neither competitive yet with traditional oil crops nor with fossil fuel. The use of renewable technologies as photovoltaics and biogas self production might increase the competitiveness of micro-algae oil. Further investigations are however necessary to optimize their production chain and to increase the added value of co-products.


Validation of Canadian mothers' recall of events in labour and delivery with electronic health records.
Maternal report of events that occur during labour and delivery are used extensively in epidemiological research; however, the validity of these data are rarely confirmed. This study aimed to validate maternal self-report of events that occurred in labour and delivery with data found in electronic health records in a Canadian setting. Data from the All Our Babies study, a prospective community-based cohort of women's experiences during pregnancy, were linked to electronic health records to assess the validity of maternal recall at four months post-partum of events that occurred during labour and delivery. Sensitivity, specificity and kappa scores were calculated. Results were stratified by maternal age, gravidity and educational attainment. Maternal recall at four months post-partum was excellent for infant characteristics (gender, birth weight, gestational age, multiple births) and variables related to labour and delivery (mode of delivery, epidural, labour induction) (sensitivity and specificity >85%). Women who had completed a university degree had significantly better recall of labour induction and use of an epidural. Maternal recall of infant characteristics and events that occurred during labour and delivery is excellent at four months post-partum and is a valid source of information for research purposes.


Dietary nitrate lowers ambulatory blood pressure in treated, uncontrolled hypertension: a 7-d, double-blind, randomised, placebo-controlled, cross-over trial.
Dietary nitrate has been shown to increase nitrate/nitrite levels and decrease blood pressure (BP) in multiple populations. There are few reports among hypertensives and these reports have provided conflicting evidence. We aimed to assess the effect of daily nitrate compared with placebo in subjects with uncontrolled hypertension (HTN). On day 0, hypertensives wore an ambulatory BP monitor (ABPM) for 24 h and blood was taken. Subjects were then randomised to 7-d nitrate-rich beetroot juice (NO3 -) (12·9 mmol nitrate) followed by 7-d nitrate-depleted beetroot juice (0·5 mmol nitrate) or vice versa. ABPM and blood were assessed before and after both conditions. In all, twenty subjects with treated yet uncontrolled HTN entered and completed the trial (mean age=62·5 years, mean BMI=30·7 kg/m2). Baseline BP was 137/80 (sd 7/7) mmHg. Dietary nitrate was well tolerated and resulted in significantly increased plasma nitrite (P=0·0004) and decreased 24-h systolic BP and diastolic BP compared with placebo (-8 mmHg; P=0·012 and -4 mmHg; P=0·018, respectively). Our results support the existing data suggesting an anti-hypertensive effect of dietary nitrate in treated yet uncontrolled hypertensives. Targeted dietary strategies appear promising contributors to BP control.


Subtle right ventricular dysfunction in asymptomatic chronic heavy cigarette smokers: a speckle tracking case-control study.
Chronic heavy cigarette smoking can affect the right ventriclular function. The standard echocardiography may not show early right ventricular functional changes, and a more sensitive measure is needed. The aim of this work was to evaluate the subtle subclinical effects of chronic heavy cigarette smoking on the right ventricular function. The study included 55 healthy asymptomatic chronic heavy cigarette smokers (smoking history of at least 5 pack-years and a daily cigarette consumption of at least 1 pack) and 35 healthy non-smoking control subjects. Patients underwent a full clinical assessment and a conventional as well as a 2D-speckle tracking transthoracic echocardiography of the right ventricle and data was compared between the 2 groups. The mean age was 32.9 ± 7.2 years in smokers and 30.9 ± 7.9 years in non-smokers (p = 0.227). The 2 groups showed comparable conventional right ventricular systolic and diastolic functions. Smokers showed a significantly lower (less negative) right ventricular global longitudinal strain (- 19.0 ± 3.2% vs. - 24.5 ± 3.5%, p < 0.001). Patients with a higher daily cigarette consumption showed a poorer right ventricular global longitudinal strain (p = 0.014). Chronic heavy cigarette smoking can adversely affect the right ventricular function, a finding that can be easily missed by conventional echocardiography and can be better detected by the right ventricular speckle tracking.


Polypeptides synthesized in rabbit cells infected with murine herpesvirus (MHV): a comparison of proteins specified by various MHV strains.
Polypeptides synthesized in rabbit embryo fibroblasts (REF) infected with six isolates of murine herpesvirus (MHV) coming from different localities in CSFR were compared by electrophoresis (SDS-PAGE). Altogether 28 virus-coded polypeptides with apparent molecular weights ranging from 240 kD to 16 kD were identified. While isolates MHV-60, MHV-72, and MHV-76 had identical polypeptide profiles, the isolates MHV-68 and MHV-78 lacked a polypeptide with apparent molecular weight of 46 kD, the rest of polypeptides being identical as well. The polypeptide profile of MHV Sumava A.f. was most different from that of other MHV isolates. This result is in agreement with some different biological properties of this isolate. At a high multiplicity of infection (40 TCID50 per cell) first polypeptides with apparent molecular weights 125 kD and 84 kD were detected by 5-9 hr post-infection (p.i.). All other proteins appeared 14-18 p.i.


A new affordable flow cytometry based method to measure HIV-1 viral load.
Most commercially available assays for diagnosis of HIV infection have shown shortcomings in the detection and quantification of rare genotypes of the virus. Most of the assays do not detect subtype O (outlier) and/or N (nonmajor, nonoutlier) or new circulating recombinant forms (CRFs), which are becoming more important in sub-Saharan Africa. Furthermore, the commonly available tests require costly measuring devices and expensive test kits, which are not easily affordable for developing countries. This study was designed to explore solutions to the problem of viral load assays in developing countries. Two forward primers, digoxygenin (DIG) and dinitrophenol (DNP) labeled, and one biotin (BIO) labeled reverse primer were used to amplify both, the HIV-1-5'LTR (long terminal repeat) region and an internal standard sequence. The two polymerase chain reaction (PCR)-products were captured by anti-DIG and anti-DNP antibody coated microparticles. Flow cytometric analyses were carried out after labeling with streptavidin-R-phycoerythrine. The primer system used recognized all HIV-1 subtypes. A coamplified internal standard warranted the functionality of the PCR and allows reproducible viral load measurements. Two drawbacks of current viral load measurements are overcome by the flow cytometry based test described hereof. First, all known worldwide relevant HIV-1 subtypes including subtypes O, N, and new CRFs are quantifiable with high sensitivity (50 to >1 x 10(6) copies per PCR). Second, the cost per test can be reduced to less than 12 US$ instead of the current 50-100 US$. Additionally, the test described in this report offers the possibility to perform complete monitoring program (CD4 T-cell count, CD4% and viral load) for the first time, with the same device for HIV-infected persons.


Are Confirmatory Assays Reliable for HIV-1/HIV-2 Infection Differentiation? A Multicenter Study.
Immunoblots remain the gold standard for HIV-1/HIV-2 infection confirmation. However, their ability to differentiate HIV-1 from HIV-2 infection on an antigenically diversified HIV-1 and HIV-2 panel remain uncommon. We performed a multicenter study on 116 serum samples accounting for most of the diversity of HIV-1 (9 different subtypes in group M, 17 circulating recombinant forms (CRFs), and 3 group O) and HIV-2 (groups A and B), evaluating seven confirmatory assays (six commercially available assays and one in-house assay) with genotyping as the reference. The assays were INNO-LIA HIV I/II score, HIV-2 blot 1.2, HIV blot 2.2, New Lav blot I and II, Geenius, and an in-house serotyping enzyme-linked immunosorbent assay (ELISA). Among the HIV-1 samples, INNO-LIA, HIV blot 2.2, New Lav blot I, Geenius, and serotyping had comparable high sensitivities, from 98% to 100%, whereas HIV-2 blot 1.2 and New Lav blot II had high rates of "undetermined" results (85% and 95%, respectively). HIV-2 blot 1.2 and New Lav blot II misclassified 7% and 5% of HIV-1 samples as HIV-2, respectively, and HIV-2 blot 1.2 had an 8% false-negative rate. Among the HIV-2 samples, INNO-LIA, New Lav blot II, HIV-2 blot 1.2, and serotyping had high sensitivities, from 96% to 100%. HIV blot 2.2 misclassified 17% of HIV-2 samples as HIV-1/HIV-2 dual infections. New Lav blot I misclassified 19% of HIV-2 samples as HIV-1 with a high (81%) undetermined rate, and Geenius misclassified 2% as HIV-1 and 7% as untypeable HIV positive. For HIV-1/HIV-2 dual infection, the results were less sensitive, with at most 87.5% for INNO-LIA and Geenius and 75% for HIV blot 2.2 and serotyping. Overall, confirmatory assays remain useful for most cases, with the exception of HIV-1/HIV-2 dual-infection suspicion.


Apigenin ameliorates oxidative stress and mitochondrial damage induced by multiwall carbon nanotubes in rat kidney mitochondria.
The toxicity of carbon nanotubes (CNTs) toward the mitochondria of the kidney is not fully recognized and still needs further research. Apigenin (APG) is known as a flavonoid compound and natural antioxidant. The purpose of this study was to assess the ameliorative role of APG against multiwall CNT (MWCNT)-induced kidney toxicity in rats. The animals were administrated with APG (10 mg/kg) for 2 weeks and then were exposed to MWCNTs (5 mg/m3 ) in pure and impure forms (10 and 100 nm) for 5 h/day and 5 days/week. Then, mitochondria were isolated from the kidney tissue and mitochondrial toxicity parameters were measured. Decreases in succinate dehydrogenase activity have been reported in all groups exposed to MWCNTs. Results indicated that MWCNTs in both forms and sizes were able to increase the generation of reactive oxygen species, decline mitochondrial membrane potential, induce mitochondrial swelling, and release cytochrome c in isolated kidney mitochondria. The pretreatment of APG decreased all the abovementioned mitochondrial damage and oxidative stress parameters induced by both pure and impure MWCNTs. Our results showed that MWCNTs have the ability to enter the body, subsequently, cross cellular barriers, and reach the kidney as a sensitive organ, which can result in mitochondrial damage in kidney cells including renal tubular cells. In addition, APG can be an effective nutritional antioxidant regimen against MWCNT-induced kidney damage.


Benzodiazepine receptors studied in living primates by positron emission tomography: inverse agonist interactions.
The convulsant actions of methyl 6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM) and of methyl beta-carboline-3-carboxylate (beta-CCM) were evaluated in the baboon (Papio papio). DMCM, 0.6-4 mg/kg, induced epileptic seizures with short latency. DMCM convulsive seizures could be blocked by i.v. administration of the benzodiazepine agonist diazepam (10 mg). Similarly, beta-CCM, 0.3-3 mg/kg i.v., provoked generalized seizures in the baboons. These seizures were also reversed by the administration of propyl beta-carboline-3-carboxylate (3 mg/kg) or of diazepam (5 mg/kg). Combining the results from Positron Emission Tomography and the EEG assessments, benzodiazepine receptor occupancy by beta-CCM and DMCM was directly correlated with their convulsant actions in the living baboon. beta-CCM exerted its convulsant action in the living baboon at 76 and 74% benzodiazepine receptor occupancy in, respectively, occipital and temporal cortices whereas DMCM displayed a similar convulsive activity when only 58 and 65% of these receptors in the above regions were occupied.