Health and doping in elite-level cycling.
The protection of the health of athletes is one of the three criteria taken into account when registering a substance in the World Anti-Doping Agency prohibited list. Nevertheless, in elite-level cycling, banned substance use is widespread. The present research adopted a psychological approach to examine how or whether perceived health risks influence elite-level cyclists' decisions to use banned substances. Sixteen semi-structured interviews were conducted with cyclists hoping to join a professional team (n=6), neo-professional cyclists (n=2), and former professional cyclists (n=8). Although an evolution was observed in the organization of doping and perceptions of doping over the last decade, the perceived health hazards did not influence, most of the time, decisions to use banned substances among the sample of cyclists. There was a systematization of exogenous substance use in the cycling environment and a trivialization of the side effects of the banned substances. Finally, younger cyclists were not concerned about the long-term health consequences of banned substances; they were more focused on the short-term performance-enhancing benefits. There is a need to implement more effective preventive programs to change athletes' attitudes toward doping and its health risks.


A biomechanical analysis of patellofemoral stress syndrome.
THIS STUDY WAS CONDUCTED IN ORDER TO: a) investigate the relationship of selected anthropometric, strength, and kinematic variables to the incidence of patellofemoral stress syndrome in high school female athletes; and b) develop a predictive equation to screen individuals who may be predisposed to patellofemoral stress syndrome. Twenty-nine subjects were analyzed across nine dependent variables: two anthropometric measures, one strength measure, and six kinematic measures. Heavy subjects and those with a larger static quadriceps angle (Q-angle) were more likely to have patellofemoral stress syndrome. Leg strength did not seem to be a factor. Also, a variable of gait, the time from foot contact to the time when the minimum dynamic Q-angle occurred, was significantly slower in the subjects with patellofemoral stress syndrome. Furthermore, a predictive equation, which we created using discriminant analysis, was 89% accurate in predicting which subjects would or would not have patellofemoral stress syndrome. The equation uses an individual's weight, pelvic width, and static Q-angle. We conclude that, through proper screening, individuals susceptible to patellofemoral stress syndrome may be identified prior to their becoming symptomatic, and that, through identifying causal variables, corrective procedures may be introduced in order to prevent patellofemoral stress syndrome from hindering an individual's physical activity.


A phase I randomized safety study of a single-size silicone rubber diaphragm used with or without a lactic-acid-containing diaphragm gel.
To evaluate a lactic-acid-containing diaphragm gel (Contragel®) approved outside the United States for use with a silicone rubber diaphragm (Caya®). The study gel is being evaluated as a safer alternative to nonoxynol-9 (N-9) gel, which has been associated with risk of increasing susceptibility to human immunodeficiency virus (HIV). This was a Phase I randomized, parallel study evaluating the safety of the novel diaphragm gel versus hydroxyethylcellulose (HEC) universal placebo gel delivered by the study diaphragm for two 7-day test cycles of daily use, without and with intercourse. The primary clinical safety endpoint was treatment emergent adverse events. Mucosal safety endpoints included colposcopic findings, anti-Escherichia coli activity of endocervical and vaginal fluid, immune mediators, Nugent score and ectocervical immune cell density. Endpoints were assessed prior to each test cycle and at day 7 of each test cycle. We compared the two independent groups and also evaluated paired changes from baseline in each gel cohort. Twenty-three participants used the study diaphragm with the novel gel (n=11) or with HEC (n=12). Use of either gel resulted in few genital AEs and no colposcopic findings. There were no differences in ectocervical histology and lymphocyte density or phenotype between the two cohorts at baseline or after each test cycle. We found no clinically important differences in the anti-microbial (anti Escherichia coli) activity of endocervical or vaginal fluid or concentrations of genital immune mediators (e.g. anti-inflammatory secretory leukocyte protease inhibitor (SLPI) or pro-inflammatory mediator RANTES) between the two gel cohorts at any visit. There were no important paired changes from baseline among participants using either gel in Nugent score, ectocervical histology or anti-microbial activity of genital secretions. We found no clinically significant differences in clinical and mucosal safety endpoints between the two cohorts. The mucosal safety profiles of the study gel and HEC placebo gel were similar. Our data demonstrate no clinically important differences between the safety profiles of the lactic-acid-containing diaphragm gel versus HEC placebo gel when used with the study diaphragm. N-9 can no longer be used with contraceptive diaphragms in high HIV prevalence regions. Although larger studies are needed, the novel gel appears safe for use with the study diaphragm, which is the first over-the-counter, non-hormonal, diaphragm.


Assignment of Clostridium bryantii to Syntrophospora bryantii gen. nov., comb. nov. on the basis of a 16S rRNA sequence analysis of its crotonate-grown pure culture.
The Clostridium bryantii-Methanospirillum hungatei syntrophic coculture, grown on caproate, was adapted to grow on crotonate. Then, C. bryantii was isolated in pure culture from crotonate bottle plates. A 16S rRNA sequence analysis of the pure subculture revealed that, as a member of the gram-positive phylum, it was not closely related to any of the Clostridium species with which it was compared or to any of the other clusters in the gram-positive phylum with which it was compared. However, it was closely related to another syntrophic fatty acid-degrading bacterium, Syntrophomonas wolfei. On the basis of its phylogeny, physiology, and cell wall ultrastructure, we propose assignment of C. bryantii to Syntrophospora bryantii gen. nov., nov. comb.


Parental imagery vividness: gender differences and relationships to emotional closeness.
This research was an investigation of the vividness with which individuals see their parents in different situations. In 3 studies, the Vividness of Visual Imagery for Parents Questionnaire (VVIPQ; S. J. McKelvie, 1998a) was completed by university undergraduates (201 women, 167 men). Visual imagery of mothers was reported to be more vivid than that of fathers, but only by the female students. For intact families, the female students also reported more imagery of mothers than of fathers, whereas the male students reported more vivid imagery of fathers than of mothers. However, for divorced or separated families in which individuals spent more time with their mothers, both men and women rated their mothers more vividly than their fathers. More vivid imagery was positively related to reported emotional closeness to parents and to more vivid imagery for general scenes. VVIPQ scores were associated with social desirability but were not related to measures of response leniency or general intelligence. Split-half reliability of the VVIPQ was very good, and both test-retest and alternate-format reliability were acceptable. The correlational evidence supports the construct validity of the VVIPQ, which is a promising tool for studying parental imagery.


Daily multivitamins with iron to prevent anemia in high-risk infants: a randomized clinical trial.
The goal of this study was to assess the effectiveness of multivitamins with iron as prophylaxis against iron deficiency and anemia in infancy. The study was a double-blinded, randomized, pragmatic, clinical trial conducted at 3 urban primary care clinics. Subjects included healthy, full-term infants who were enrolled at their 6-month well-child visit. Infants were randomly assigned to receive standard-dose multivitamins with or without iron (10 mg/day). Parents administered multivitamins by mouth daily for 3 months. Laboratory results at 9 months of age were analyzed for the presence of anemia and/or iron deficiency. Anemia was defined as hemoglobin level <11.0 g/dL. Iron deficiency was initially defined as any abnormal laboratory value of the following: mean corpuscular volume combined with red cell distribution width or zinc protoporphyrin (with blood lead level <10 microg/dL) for most subjects and ferritin, transferrin saturation, or reticulocyte hemoglobin content for a subset. Subsequent analyses defined iron deficiency as any 2 abnormalities of the above laboratory outcomes, except hemoglobin. The control (n = 138) and intervention (n = 146) groups were equivalent with respect to all important sociodemographic and nutritional variables. At 9 months of age, anemia was found in 21% of infants (n = 58). A total of 229 (81%) had iron deficiency on the basis of 1 abnormal laboratory indicator and 139 (49%) on the basis of 2 abnormal laboratory indicators. No difference existed in the occurrence of anemia and iron deficiency between the intervention and control groups. In the intervention group, 22% and 78% of 138, respectively, were anemic or had 1 abnormal laboratory outcome indicative of iron deficiency. In the control group, 19% and 84% of 144 were anemic or iron deficient. When stratified by adherence, no differences in hematologic outcomes between groups were noted. However, in multivariate logistic regression, infants whose mothers were anemic during pregnancy were 2.15 times more likely than others to have any laboratory abnormality (95% confidence interval: 1.14-4.07). Increasing adherence, regardless of group assignment, was associated with a 0.56 times reduced risk of any abnormality (95% confidence interval: 0.41-0.76). On the basis of intention-to-treat analysis, multivitamins with iron was not effective in preventing iron deficiency or anemia in 9-month-old infants. However, effective prevention and treatment of maternal anemia during pregnancy and giving multivitamins with or without additional iron during infancy may prove to be important approaches to the prevention of iron deficiency among high-risk children. Because of the consequences of iron deficiency and its high prevalence among low-income infants, additional investigation in these areas is warranted.


Theory for Modeling of High Resolution Resonant and Nonresonant Raman Images.
Tip-enhanced Raman imaging is capable of resolving the inner structure of a single molecule owing to the generation of highly localized nanocavity plasmon. Here we present a general theory and detailed computational methodology to fully describe resonant and nonresonant Raman scattering under the localized plasmonic field. We use an allylcarbinol molecule adsorbed on the gold surface as a model system to illustrate different effects on the Raman images. It is found that the ability of distinguishing an individual vibration mode is highly limited under the resonant condition due to the dominant contribution from the Franck-Condon term and the mode-independent component of the Herzberg-Teller term. The nonresonant Raman images of the single molecule are vibrationally distinguishable and present the vibrational motion of the corresponding vibrational modes in real space. Furthermore, the calculated results confirm that nonlinear optical effects can further improve the resolution of the images. The theoretical and computational methods presented here provide the basic tools to model high resolution Raman images at the single molecular level.


Zinc-dependent structure of a single-finger domain of yeast ADR1.
In the proposed "zinc finger" DNA-binding motif, each repeat unit binds a zinc metal ion through invariant Cys and His residues and this drives the folding of each 30-residue unit into an independent nucleic acid-binding domain. To obtain structural information, we synthesized single and double zinc finger peptides from the yeast transcription activator ADR1, and assessed the metal-binding and DNA-binding properties of these peptides, as well as the solution structure of the metal-stabilized domains, with the use of a variety of spectroscopic techniques. A single zinc finger can exist as an independent structure sufficient for zinc-dependent DNA binding. An experimentally determined model of the single finger is proposed that is consistent with circular dichroism, one- and two-dimensional nuclear magnetic resonance, and visual spectroscopy of the single-finger peptide reconstituted in the presence of zinc.


Lower respiratory tract myeloid cells harbor SARS-CoV-2 and display an inflammatory phenotype.
SARS-CoV-2 pneumonia may induce an aberrant immune response with brisk recruitment of myeloid cells into the lower respiratory tract, which may contribute to morbidity and mortality. We describe endotracheal aspirate samples from seven patients with SARS-CoV-2 pneumonia requiring mechanical ventilation. We note SARS-CoV-2 virions within lower respiratory tract myeloid cells shown by electron tomography, immunofluorescence confocal imaging, and immuno-electron microscopy. Endotracheal aspirates are primarily composed of mononuclear and polymorphonuclear leukocytes. These myeloid cells that harbor virus are frequently positive for CD14 and/or CD16 and most display an inflammatory phenotype marked by expression of IL-6 and tissue factor mRNA transcript and protein expression.


New epidemiology of hepatitis delta.
Hepatitis D virus (HDV) is a defective pathogen that needs hepatitis B virus (HBV) for infection. Co-infection of HBsAg-positive individuals with HDV is commonly associated with a more rapid progression to cirrhosis, a higher incidence of hepatocellular carcinoma (HCC) and increased mortality. Initial studies have shown that about 5% of chronic HBV carriers worldwide (15-20 millions) were also infected with HDV. However, recent studies suggest that the prevalence of HDV is at least two- to three-fold higher than previous estimations. Improved diagnostic techniques have shown that HDV infection remains endemic in certain areas of the world. Injection drug users, individuals with high-risk sexual behaviour and patients co-infected with human immunodeficiency virus (HIV) represent the major reservoir of the disease in the Western world. Although the burden of HDV infection significantly decreased in Europe in the nineties, there has been no further decrease in the last decade, probably because of migration from HDV endemic countries. Until new and more effective therapies are available, public health measures should be reinforced by increasing prophylactic HBV vaccination programs, preventing transmission of the virus among parenteral drug users and implementing universal HDV screening of all HBV-infected individuals.


Thymoma Causing Bilateral Upper Extremity Deep Vein Thrombosis.
A 38-year-old African American male presented with progressive pain, swelling, numbness, and warmth of the left upper extremity ten days before admission. A chest computerized tomography scan showed a large 8.3 cm × 6.1 cm x 9.9 cm anterior mediastinal mass with compression of the left brachiocephalic vein and superior vena cava. A venous doppler showed multiple occlusive venous thrombi in bilateral upper extremities, including the bilateral internal jugular and subclavian veins, as well as the left subclavian, axillary, cephalic, brachial and median cubital veins. Further laboratory workup came positive for acetylcholine receptor binding antibody suggesting myasthenia gravis, but the patient was asymptomatic for myasthenia gravis. A percutaneous core CT guided biopsy pathology resulted in a predominant T-cell population CD5 positive with few B cells; the immunophenotypic features suggested Type B2 thymoma. To the best of our knowledge, this case is the only reported thymoma presenting with bilateral deep vein thrombosis of the upper extremities. The deep vein thrombosis therapy was enoxaparin 1mg/kg subcutaneously every 12 hours and dexamethasone 4mg intravenously every 4 hours as an anti-inflammatory drug for thymoma related compression of the mediastinum. The patient was referred to a tertiary oncological medical center for a total thymectomy, chemotherapy, and adjuvant radiotherapy.


Determination of Depth in Transmission Raman Spectroscopy in Turbid Media Using a Beam Enhancing Element.
This study demonstrates experimentally a method to enable prediction of depth of a chemical species buried in a turbid medium by using transmission Raman spectroscopy alone. The method allows the prediction of the depth of a single, chemically distinct layer within a turbid matrix by performing two measurements, with and without a beam enhancing element, or "photon diode." The samples employed consisted of two different polymers, of total thickness 3.6 mm, whose optical properties are loosely relevant to pharmaceutical applications. A polymer layer of low-density polyethylene (LDPE) was placed at different positions within multiple layers of the polytetrafluoroethylene (PTFE) matrix and Raman spectra were recorded in each case. Both univariate and multivariate analyses were utilized to determine whether the depth of the LDPE layer could be predicted using the obtained data. The best-achieved RMSE of prediction was 4.2% of the total sample size (i.e., +/- 0.15 mm) with the multivariate approach.


Impact of Frailty on Bleeding Events Related to Anticoagulation Therapy in Patients With Atrial Fibrillation.
Although anticoagulation is the key treatment to prevent stroke in patients with atrial fibrillation (AF), including elderly patients, anticoagulation is sometimes withheld for elderly people because of concerns about frailty. However, it remains unknown whether frailty increases bleeding events.Methods and Results:A total of 120 consecutive non-valvular AF patients admitted with symptoms of AF or congestive heart failure were included in this study. Frailty was assessed using the Cardiovascular Health Study (CHS) frailty index. We performed a retrospective analysis of the risk factors associated with major bleeding events. After a median follow-up of 518 days, major bleeding events occurred in 17 (14.2%) patients. Patients with major bleeding events had a higher CHS frailty index (P=0.015). The cutoff value for high-risk CHS frailty index was 2 (area under the ROC curve: 0.68 [95% confidence interval (CI): 0.57-0.78]). The event-free rates at 2 years were 97.6% (95% CI: 83.9-99.7) in patients with a CHS frailty index <2 and 59.6% (95% CI: 27.9-81.0) for those with a CHS frailty index ≥2 (P<0.001). Frailty is associated with increased bleeding events related to anticoagulant therapy in patients previously hospitalized with AF. Greater care should be taken with patients with a CHS frailty index ≥2.


A novel implementation of signature, encryption and authentication (SEA) protocol on mobile patient monitoring devices.
Mobile patient monitoring devices are becoming an integral part of healthcare industry and these devices will eventually become the method of choice for accessing and implementing health checks for patients located in remote areas. The thrust behind this research work was to investigate how a complete security strategy, comprising of digital signatures, encryption and authorisation on the mobile healthcare devices could be implemented without compromising overall system performance. Java 2, Micro Edition (J2ME) platform has been used to implement the Signature, Encryption and Authentication (SEA) protocol based on Elliptic Curve Cryptography (ECC). The work presented highlights a practical, secure and effective prototype for interactive monitoring of remote patients. The proposed facilities could improve the quality of care and doctor-patient communications; prevent misuse of healthcare data and consequential expenses to the healthcare industry in an efficient user friendly way that does not significantly affect the device performance.


Role of the sperm, oocyte, and embryo in recurrent pregnancy loss.
Disorders affecting the sperm, oocyte, or embryo may cause a significant fraction of spontaneous miscarriages and cases of recurrent pregnancy loss (RPL). Altered chromosomal integrity of sperm and oocytes, which is highly dependent of the age of the mother, represents a major cause of miscarriage and in turn RPL. Avoiding transfers of abnormal embryos is possible with preimplantation genetic testing for aneuploidies. Chromosomal anomalies may also be caused by structural rearrangements of one or several chromosomes in either parents, a finding encountered in 12% of couples with RPL, including in those who have had one or several healthy babies. More than 40% of these chromosomal rearrangements are identifiable on regular karyotypes. When abnormal findings are made, preimplantation genetic testing for monogenic disorders allows selection of disease-free embryos. Finally, asymmetric inactivation of the X chromosome has been found more commonly in women with RPL, but no specific treatment is currently available.


Small-molecule inhibitor BMS-777607 induces breast cancer cell polyploidy with increased resistance to cytotoxic chemotherapy agents.
The RON receptor tyrosine kinase is a therapeutic target for cancer treatment. Here, we report therapeutic effect and phenotypic change of breast cancer cells in response to BMS-777607, a RON tyrosine kinase inhibitor. Treatment of breast cancer cells with BMS-777607 at therapeutic doses inhibited cancerous clonogenic growth but had only minimal effect on cell apoptosis. Significantly, BMS-777607 induced extensive polyploidy with multiple sets of chromosomes in cancer cells. This effect is independent of RON expression. Knockdown of RON in T-47D and ZR-75-1 cells by specific siRNA did not prevent polyploid formation. Immunofluorescent analysis of α-tubulin and γ-tubulin expression in polyploid cells revealed that BMS-777607 disrupts bipolar spindle formation and causes multipolar-like microtubule assembly. Also, both metaphase equatorial alignment and chromosomal segregation were absent in polyploid cells. These results suggest that cellular mitosis arrests at prophase/pro-metaphase and fails to undergo cytokinesis. By analyzing kinase-inhibitory profiles, aurora kinase B was identified as the target molecule inhibited by BMS-777607. In BMS-777607-treated cells, aurora kinase B was inhibited followed by protein degradation. Moreover, BMS-777607 inhibited Ser10 phosphorylation of histone H3, a substrate of aurora kinase B. Chemosensitivity analysis indicated the resistance of polyploid cells toward chemotherapeutics. Treatment with doxorubicin, bleomycin, methotrexate, and paclitaxel significantly increased cellular IC50 values. These findings highlight the theory that BMS-777607 acts as a multikinase inhibitor at therapeutic doses and is capable of inducing polyploidy by inhibiting aurora kinase B. Increased resistance of polyploid cells to cytotoxic chemotherapeutics could have a negative impact on targeted cancer therapy using BMS-777607.


Sex-Based Differences in Melanoma Survival in a Contemporary Patient Cohort.
Background: A female survival advantage in cutaneous melanoma has been long recognized. However, whether this extends across all age groups, with risk stratification using the latest prognostic staging system or in the current era of efficacious systemic therapies is unknown. Therefore, we evaluated whether sex-based differences in melanoma survival persisted within a recent population-based patient cohort with consideration of these factors. Materials and Methods: We identified stage II-IV cutaneous melanoma patients from 2010 to 2014 Surveillance, Epidemiology, and End Results cancer registries data. We recalculated stage per American Joint Committee on Cancer 8th edition guidelines. Cancer-specific survival (CSS) was estimated by using the Kaplan-Meier method and multivariable Cox proportional hazards regression. Results: Of 16,807 patients (39.8% female), 8,990 were stage II, 4,826 stage III, and 2,991 stage IV at diagnosis. Unadjusted 3-/5-year CSS estimates for females versus males were 64.2% versus 59.7%, and 53.5% versus 49.9%, respectively, p ≤ 0.0001. Five-year CSS varied within each stage and across age strata of <45, 45 - 59, and ≥60 years. Within each stage, females <45 had better CSS than all other sex/age groups (p < 0.0001). In multivariable analysis of stage II/III patients, female sex, younger age, and lower mitotic index retained favorable CSS prognostic significance (p < 0.001). Conclusions: Sex-based differences in melanoma survival persist in a contemporary patient cohort staged with the latest prognostic system. These data may guide decision marking regarding adjuvant therapy, highlight the importance of including sex as a pre-specified clinical trial variable, and suggest that investigation of underlying biologic mechanisms may drive discovery of biomarkers and therapeutic targets to improve patient care.


Geographical region traceability of wild topmouth culter (Culter alburnus) from Xingkai Lake based on muscle quality and aroma profiles.
The wild topmouth culter (Culter alburnus) from Xingkai Lake (XKL) is highly regarded for its delicious taste and unique flavor. In this study, based on muscle quality and aroma analysis, we first differentiated the XKL population from three wild populations in Heilongjiang Province and one artificially cultured population (from Xingkai Lake). Compared with the other populations, the XKL population has a significantly higher crude protein content, essential amino acid content, delicious amino acid content, and n-3/n-6 PUFA ratio. Additionally, it exhibits superior hardness, elasticity, chewiness, recoverability, and viscosity. E-nose detection analysis revealed that W1S, W2S, and W3S were the potential sensors contributing the most to the differences among the five populations. HS-SPME-GC-MS and multivariate regression analysis showed that 21 volatile flavor compounds were identified as key markers for geographical identification of the Xingkai Lake region. These findings will provide guidance for the geographical traceability and identification of the XKL population.


Electrically detected magnetic resonance using radio-frequency reflectometry.
The authors demonstrate readout of electrically detected magnetic resonance at radio frequencies by means of a LCR tank circuit. Applied to a silicon field-effect transistor at millikelvin temperatures, this method shows a 25-fold increased signal-to-noise ratio of the conduction band electron spin resonance and a higher operational bandwidth of >300 kHz compared to the kilohertz bandwidth of conventional readout techniques. This increase in temporal resolution provides a method for future direct observations of spin dynamics in the electrical device characteristics.


[The professional burnout syndrome in resident physicians in hospital medical specialties].
To evaluate the existence of burnout among medicine residents of hospitality specialties, and the relationship with sundry factors. A transversal study was made among internal medicine residents of La Fe Hospital with more than one year in their job. For this purpose two questionnaires were used: a) the Maslach Burnout Inventory and a general questionnaire about social, demographic, residency, laborer and economic factors. Fifty-six (81.1%) internal medicine residents participated. Comparing with Spanish sample established with medical specialists, we found 7.1% or the residents with high scores on emotional exhaustion, while 17.8% scored high on depersonalization and 23.2% scored low on personal accomplishment. But the levels were lower on residents sample than in the medical specialties sample. Factors associated with burnout were social and demographic compass (be single), like on residency compass (the chosen specialty was not the first option), like properly laborer (low laborer satisfaction, to feel low recognition from boss or patients). We didn't find relationship with economic factors. There are hospitality internal medicine residents who suffer burnout and exist relationship with social, demographic, residency and laborer factors.


Aromatic amino acid requirement of the lactating sow.
Six mature Yorkshire X Landrace sows were randomly assigned to a 6 X 6 Latin-square experiment on d 5 of lactation to determine the total phenylalanine (Phe) and tyrosine (Tyr) requirement. A control diet of corn sugar, cornstarch, whey, L-glutamic acid, solka floc, soybean oil, amino acids, minerals and vitamins was supplemented with L-Phe to provide .30 (basal), .45, .60, .75, .90 and 1.05% total aromatic amino acids (TAAA). Each diet was fed to a maximum of 5.5 kg/d within each of six 10-d periods. Feed intake, average pig weight gain and sow milk yield decreased linearly (P less than .01) with increasing period. Sow milk yield was maximized at .75% TAAA (quadratic, P less than .10), but average pig weight gain did not reflect the higher yield. Urea nitrogen decreased quadratically with increasing dietary Phe in both plasma (P less than .05) and urine (P less than .01) to a breakpoint at .56% TAAA. Plasma Phe increased (quadratic, P less than .01) as dietary TAAA increased, but no clear inflection point was obtained. A sharp rise (quadratic, P less than .001) in plasma Tyr occurred at .73% dietary TAAA. Plasma lysine decreased (linear, P less than .001) to a low level at .76% TAAA, but plasma methionine was unaffected by treatment. Urine allantoin/urea X protein intake was maximized at .61% TAAA (quadratic, P less than .01). Nitrogen (N) intake varied among diets (quadratic, P less than .05), but fecal N was not altered by TAAA level.(ABSTRACT TRUNCATED AT 250 WORDS)


A plasmid-encoded nicotinamidase (PncA) is essential for infectivity of Borrelia burgdorferi in a mammalian host.
Borrelia burgdorferi, a spirochaete that causes Lyme borreliosis, contains 21 linear and circular plasmids thought to be important for survival in mammals or ticks. Our results demonstrate that the gene BBE22 encoding a nicotinamidase is capable of replacing the requirement for the 25 kb linear plasmid lp25 during mammalian infection. Transformation of B. burgdorferi lacking lp25 with a shuttle vector containing the lp25 gene BBE22 (pBBE22) restored infectivity in mice to a level comparable to that of wild-type Borrelia. This complementation also restored the growth and host adaptation of lp25-B. burgdorferi in dialysis membrane chambers (DMCs) implanted in rats. A single Cys to Ala conversion at the putative active site of BBE22 abrogated the ability of pBBE22 to re-establish infectivity or growth in DMCs. Additional Salmonella typhimurium complementation studies and enzymatic analysis demonstrated that the BBE22 gene product has nicotinamidase activity and is most probably required for the biosynthesis of NAD. These results indicate that some plasmid-encoded products fulfil physiological functions required in the enzootic cycle of pathogenic Borrelia.


[Problem of candidiasis in surgery].
On the basis of personal investigations and literature data the authors analyse etiopathogenesis of Candidiasis. The suppressed immune system due to administration of corticosteroids, antibiotics, immunodepressive drugs, the presence of the initial disease and operation proper are responsible for the increased amount of fungi on mucous membranes and their more frequent colonizations.


Does Referral Bias Impact Outcomes of Surgery for Degenerative Mitral Valve Disease?
Geographic origin is cited as a possible factor influencing outcomes of operation for repair or replacement of degenerative mitral valve (MV) disease. Our study aimed to identify the potential impact of referral bias on clinical outcomes of MV surgery. We analyzed clinical and echocardiographic information of 2353 patients undergoing primary or secondary MV surgery for degenerative MV disease. Patients were grouped as local (in-state), regional (5 surrounding states), or national referrals. The number of patients (local, 827; regional, 809; national, 717) and median follow-up time (9.1 years) were similar between geographic groups. More comorbidities were found in the local patient group. Overall operative risk was 0.7% and was greater in local and regional patients compared with national patients (0.7% and 1.1% vs 0.1%, P = .05). Valve repair was performed in 97% of isolated MV surgeries, and repair rate was similar in the 3 geographic groups. The 3 groups had similar incidences of major morbidity, but local and regional groups had higher 30-day readmissions. In univariate analysis, survival was improved in national and regional patients compared with local patients; however in multivariable analysis this difference was no longer significant. There were important variations in baseline demographic and clinical characteristics between referral groups; local and regional patients presented with more comorbid conditions compared with national referrals. Aside from a small difference in perioperative mortality, early outcomes were generally similar. Late survival, however, was superior in national patients, and this referral bias is explained by fewer associated medical illnesses.


[Dependence of the evoked potentials of the auditory cortex on the parameters of acoustic stimulus].
Experiments were conducted on 40 rabbits; a study was made of the dependence of the amplitude and the latent period of the auditory evoked potentials on the intensity, duration, accretion front, frequency and the rhythm of the acoustic stimuli (the intervals between them). The intensity of the stimuli, the accretion front and the interstimulus intervals proved to influence the induced potential parameters.


[Stromal tumor of the rectum and leiomyosarcoma of the prostate as a cause of urinary syndrome attributable to BPH].
To discuss the classifications of patients with lower urinary tract symptoms ascribed to BPH, in order to define the indication for treatment for patients with similar features but of a different etiology. The literature is also reviewed. Two patients, aged 62 and 63 years, diagnosed as having BPH were found to have a gastrointestinal stromal tumor (GIST) and a leiomyosarcoma of the prostate, respectively. The first patient had undergone retropubic resection. Frozen section analysis revealed a GIST. The second patient had undergone TURP twice. A perineal biopsy at the last medical examination showed a leiomyosarcoma of the prostate. This patient finally underwent pelvic tumorectomy. The international classification of BPH is discussed: PQSF (P: prostatic weight determined by transrectal US or DRE; Q: quality of life assessment; S: international prostate symptom score (IPSS); F: maximum urinary flow rate by uroflowmetry--Qmax). The classifications are practical, but should be improved. In our view, details useful in orienting the diagnosis could be lost by oversimplification.


Root relative water content is a potential signal for impending mortality of a subtropical conifer during extreme drought stress.
Adaptation to future climates characterized by more frequent severe droughts requires enhanced mechanistic understanding of tree mortality. However, our knowledge of the physiological limits to withstand extreme drought, and how the coordination between water and carbon traits enhances survival, is still limited. Potted seedlings of Pinus massoniana were dehydrated to three target droughts (percentage loss of stem hydraulic conductivity of ca. 50%, 85%, and 100%; PLC50 , PLC85 and PLC100 ) and then relieved from these target droughts by fully rewatering. Predawn and midday water potentials (Ψ), relative water content (RWC), PLC and nonstructural carbohydrates (NSC) were monitored. During drought, Ψ and RWC declined as PLC increased. Root RWC declined more rapidly than other organ RWCs, particularly after PLC50 stress. All organ NSC concentrations were above predrought values. During rewatering, water trait recovery declined as drought increased, with no mortality at PLC50 but 75% mortality at PLC85 . The observed stem hydraulic recovery at PLC50 following rewatering was not correlated to NSC dynamics. Collectively, our results highlighted the primary role of hydraulic failure in Pinus massoniana seedling mortality by assessing mortality threshold and links among water status and water supply. Root RWC can be considered as a potential warning signal of P. massoniana mortality.


Umbelliferone Prevents Lipopolysaccharide-Induced Bone Loss and Suppresses RANKL-Induced Osteoclastogenesis by Attenuating Akt-c-Fos-NFATc1 Signaling.
Excessive bone resorption plays a central role in the development of inflammatory bone diseases, including osteoporosis and rheumatoid arthritis. Thus, identification of agents that can effectively suppress excessive osteoclast formation and function is crucial for the prevention and treatment of inflammatory bone loss. Umbelliferone (Umb), a derivative of coumarin, is a natural bioactive compound with anti-inflammatory and antioxidant properties. However, the effect of Umb on metabolic bone diseases is unknown. In this study, we found that Umb exhibited a strong inhibitory effect on lipopolysaccharide (LPS)-induced inflammatory bone loss in vivo. Histological analysis confirmed that Umb prevented trabecular bone matrix degradation and osteoclast formation in bone tissue. In addition, Umb suppressed RANKL-induced osteoclast differentiation and abrogated bone resorption. We found that the anti-osteoclastic and anti-resorptive activities of Umb are mediated via suppression of the RANKL-induced Akt-c-Fos-NFATc1 signaling pathway and the attenuation of osteoclast-specific genes, such as TRAP, OSCAR, ATP6v0d2, and CtsK. In particular, Umb downregulated the stability of c-Fos and NFATc1 proteins, but did not suppress the expression of their mRNAs. These results indicate that Umb may be a potential therapeutic agent for inflammatory bone diseases associated with abnormal osteoclast formation and function.


False negative diagnoses of critical congenital heart disease with screening neonatal pulse oximetry.
False negative pulse oximeter results occur in new born infants with critical congenital heart disease who have an oximeter saturation ≥95%. Some of these infants have abnormal physical findings but others do not. To determine the causes of false negative results. Mathematical analysis of determinants of arterial oxygen saturation and discussion of oximeter bias. False negative oximeter results are not rare; the sensitivity of pulse oximetry screening for critical congenital heart disease is about 80%. The high saturation may be due to a very small right-to-left shunt at the time of study, a relatively high cardiac output and mixed venous saturation, or to positive bias in oximeter readings. It may also be due to some critical congenital heart lesions that do not show desaturation at the time of testing. A diagnosis of a normal heart based on a negative oximeter test is presumptive, and requires careful follow-up for 1-2 weeks after birth.


In planta Activity of the Novel Copper Product HA + Cu(II) Based on a Biocompatible Drug Delivery System on Vine Physiology and Trials for the Control of Botryosphaeria Dieback.
The growing concerns on human and environment health are forcing the plant protection industry toward the formulation of more eco-sustainable plant protection products (PPP), both efficient and innovative in their approach to disease control. A large number of these innovative formulations now rely on a combination of pathogens antagonistic properties and stimulation of natural plant defense to pathogens. The formulation HA + Cu(II), in which copper is delivered to the plants by the drug-delivery molecule hydroxyapatite (HA), was found efficient against the grapevine pathogens Plasmopara viticola and Phaeoacremonium minimum and able to induce the host-plant defense system. We investigated the HA + Cu(II) impacts on grapevine physiology, both in uninfected and when infected by the Botryosphaeria dieback agents Diplodia seriata and Neofusicoccum parvum. This study of plant physiology and disease impact were addressed to evaluate both the HA + Cu(II) potential as a plant defense elicitor and its possible and future use as PPP in vineyard. Our results showed that HA + Cu(II) induced several key-defense genes without negatively affecting plant growth and photosynthetic activity. In addition, fungistatic effect on the two Botryosphaeriaceae at the in planta tested concentrations is reported. Altogether, our results obtained under controlled conditions fully support the potential of HA + Cu(II) as a promising PPP toward grapevine trunk diseases in vineyard.


Successful engraftment of allogeneic peripheral blood stem cell transplant after nonmyeloablative preparative regimen with cytarabine and cyclophosphamide: report of 2 cases.
We report our experience with allogeneic peripheral blood stem cell transplantation (allo-PBSCT) following a nonmyeloablative conditioning regimen consisting of cytarabine (8 g/m2) and cyclophosphamide (120 mg/kg) in the treatment of 2 patients aged 50 and 55 years with refractory chronic myelomonocytic leukemia and chronic myeloid leukemia in accelerated phase, respectively. Our nonmyeloablative regimen was well tolerated by older patients at high risk of regimen-related toxicity by the conventional conditioning regimen but was immunosuppressive enough to achieve mixed chimerism. After allo-PBSCT, we monitored chimerism in these patients by fluorescence in situ hybridization using X- and Y-specific probes and polymerase chain reaction-based analysis of a variable number of tandem repeats. We found that full chimerism and graft-versus-leukemia (GVL) effects could be induced in these patients by donor lymphocyte infusions and withdrawal of posttransplantation immunosuppressive therapy. Our observations suggest that a nonmyeloablative conditioning regimen can establish mixed chimerism and that donor lymphocyte infusion may induce GVL effects in older patients at high risk of regimen-related toxicity.


The Facts or the Story? It Takes Both to Sensitize People About Unknown Health Hazards.
Communicating about new or unknown health risks is challenging because it requires audiences to engage with and process novel and often complex health information. This study examines how texts can convey awareness and increase knowledge about health risks people are unaware of. The focus is on how text genre (narrative, expository, and mixed-genre) affects relevant emotional (arousal, transportation) and cognitive outcomes (knowledge and risk severity), measured using both online (electrodermal activity) and offline self-report measures. Mixed-effects model analyses revealed that narrative texts exhibit the highest self-reported arousal, transportation, and risk severity. Additionally, transportation mediates the relationship between text genre and risk severity. Ultimately, mixed-genre texts produced significantly higher arousal peaks and confidence ratings on knowledge posttests compared to expository texts. Taken together, the findings suggest that narrative texts perform better at raising awareness, whereas mixed-genre texts seem more effective in learning. The implications for health risk communication are discussed.


Changing profile of elderly patients undergoing coronary bypass surgery.
Are elderly patients now undergoing CABG at higher risk than several years ago? 1536 patients, aged between 70 and 77 years, who underwent CABG between January 1987 and December 2001, were analysed. Group A 1987-1989 (n=177); group B 1990-1992 (n=243); group C 1993-1995 (n=362); group D 1996-1998 (n=418); and group E 1999-2001 (n=336). The Euroscore evaluated operative risk. There was an increase in the percentage of the study population from 12.5 to 24.1% (p=0.000). Mean age increased from 72.1±1.8 to 73.2±1.9 years (p=0.047). The percentage of patients with neurological (p=0.002), renal (p=0.013) and lung disease (p=0.04), a previous PTCA (p=0.000), left main stenosis (p=0.003), impaired ventricular function (p=0.000) and reoperations (p=0.01) increased. Emergency/urgent operations (p=0.001) decreased. Hospital mortality decreased from 7.3 to 5.7% (p=0.34). Only neurological problems increased significantly (p=0.03). The calculated operative risk by Euroscore remained stable (p=0.28). To eliminate the influence of the urgent/emergency situation, the Euroscore was recalculated, supposing that all patients were elective. At that moment a significant increase in the operative risk was seen (p=0.02). Over the last few years there has not only been an increase in the number of older patients undergoing CABG, but even in this older population there is an increasing number of high-risk patients.


IB-367 pre-treatment improves the in vivo efficacy of teicoplanin and daptomycin in an animal model of wounds infected with meticillin-resistant Staphylococcus aureus.
Antimicrobial peptides are known as immunomodulators and antibiotic enhancers. We report that administration of an antimicrobial peptide, IB-367, was efficacious in increasing the antimicrobial activity of daptomycin and teicoplanin in a mouse model of wound infection caused by meticillin-resistant Staphylococcus aureus (MRSA). Mice were assigned to seven groups: an IB-367 pre-treated group with no antibiotics given after challenge, two IB-367 pre-treated groups plus daptomycin or teicoplanin given after challenge, two groups treated with daptomycin or teicoplanin only after challenge, and two control groups without infection or that did not receive any treatment. The main outcome measures were quantitative bacterial culture and analysis of natural killer (NK) cytotoxicity and leukocyte phenotype. The wound, established through the panniculus carnosus muscle of mice, was infected by MRSA. Bacterial cultures of mice receiving antibiotics alone showed a -2 log decrease, whilst those for IB-367 plus daptomycin or teicoplanin showed a -4 log decrease. IB-367 plus daptomycin showed the highest efficacy. The higher antimicrobial effect exerted by IB-367 was associated with increased levels of NK cytotoxicity but not of NK cell number. IB-367 increased the number of both CD11b and Gr-1 cells 3 days after MRSA challenge, whereas both of these leukocyte populations were reduced at 10 days after challenge. Our data suggest that a combination of IB-367 with antibiotic exerts a therapeutic effect and may be useful for the management of staphylococcal wounds.


An integrative review on methodological considerations in mental health research - design, sampling, data collection procedure and quality assurance.
Several typologies and guidelines are available to address the methodological and practical considerations required in mental health research. However, few studies have actually attempted to systematically identify and synthesise these considerations. This paper provides an integrative review that identifies and synthesises the available research evidence on mental health research methodological considerations. A search of the published literature was conducted using EMBASE, Medline, PsycINFO, CINAHL, Web of Science, and Scopus. The search was limited to papers published in English for the timeframe 2000-2018. Using pre-defined inclusion and exclusion criteria, three reviewers independently screened the retrieved papers. A data extraction form was used to extract data from the included papers. Of 27 papers meeting the inclusion criteria, 13 focused on qualitative research, 8 mixed methods and 6 papers focused on quantitative methodology. A total of 14 papers targeted global mental health research, with 2 papers each describing studies in Germany, Sweden and China. The review identified several methodological considerations relating to study design, methods, data collection, and quality assurance. Methodological issues regarding the study design included assembling team members, familiarisation and sharing information on the topic, and seeking the contribution of team members. Methodological considerations to facilitate data collection involved adequate preparation prior to fieldwork, appropriateness and adequacy of the sampling and data collection approach, selection of consumers, the social or cultural context, practical and organisational skills; and ethical and sensitivity issues. The evidence confirms that studies on methodological considerations in conducting mental health research largely focus on qualitative studies in a transcultural setting, as well as recommendations derived from multi-site surveys. Mental health research should adequately consider the methodological issues around study design, sampling, data collection procedures and quality assurance in order to maintain the quality of data collection.


Systematic review and meta-analysis: susceptibility-guided versus empirical antibiotic treatment for Helicobacter pylori infection.
The cure rate of standard triple therapy for Helicobacter pylori infection is unacceptably low. Susceptibility-guided therapies (SGTs) have been proposed as an alternative to standard empirical treatments. The aim of this study was to perform a systematic review and meta-analysis evaluating the efficacy of SGTs. A systematic search was performed in multiple databases. Randomized controlled trials comparing cure rates of SGTs versus those of empirical therapy were selected and analysed separately for first- and second-line treatments. A meta-analysis was performed using risk ratio (RR) and number needed to treat (NNT) to measure the effect. Twelve studies were included in the meta-analysis. In first-line treatment, SGT was more efficacious than empirical 7-10 day triple therapy (RR 1.16, 95% CI 1.10-1.23, I (2) = 33%; NNT = 8). Most studies used a 7-10 day triple therapy and randomized the patients after endoscopy and/or culture, thus precluding the comparison of SGT versus non-invasive testing and empirical treatment in clinical practice. For second-line therapy, only four studies were found. Results were highly heterogeneous and no significant differences were found (RR 1.11, 95% CI 0.82-1.51, I (2) = 87%). Once endoscopy and culture have been performed, SGT is superior to empirical 7 or 10 day triple therapy for first-line treatment. Further studies are needed to evaluate the effectiveness of SGT in clinical practice, especially when compared with currently recommended first-line quadruple therapies.


[Urease biosynthesis and isolation in Staphylococcus saprophyticus L-1].
Experiments were carried out to investigate the effect of organic components of the medium and cultivation conditions on the multiplication rate and urease biosynthesis by Staphylococcus saprophyticus L-1 cells isolated from natural sources. The yeast enzymic hydrolyzate and corn extract were found to be an adequate substitute for the costly organic components--peptone and yeast extract. The substitutes ensured a high level of urease biosynthesis and biomass accumulation. The biomass accumulation was maximum at pH 6.0-7.0 and the urease activity reached maximum at pH 6.0-6.5. The optimum temperature of cultivation was 37 degrees C. Enhanced aeration and constant pH during microbial cultivation in 250 1 fermenters did not increase the biomass accumulation or urease biosynthesis as compared to flask cultivation. The study of urease isolation from the cell extract showed that the ratio of 3 volumes of ethanol to 1 volume of homogenate was optimum and provided the best precipitation of the enzyme. Preliminary thermal treatment of the cell extract increased the urease activity by 2.5 times. In this situation the activity yield was close to 100%.


CREB-independent regulation by CBP is a novel mechanism of human growth hormone gene expression.
Hypothalamic growth hormone-releasing hormone (GHRH) stimulates growth hormone (GH) gene expression in anterior pituitary somatotrophs by binding to the GHRH receptor, a G-protein-coupled transmembrane receptor, and by mediating a cAMP-mediated protein kinase A (PKA) signal-transduction pathway. Two nonclassical cAMP-response element motifs (CGTCA) are located at nucleotides -187/-183 (distal cAMP-response element; dCRE) and -99/-95 (proximal cAMP-response element; pCRE) of the human GH promoter and are required for cAMP responsiveness, along with the pituitary-specific transcription factor Pit-1 (official nomenclature, POU1F1). Although a role for cAMP-response element binding protein (CREB) in GH stimulation by PKA has been suggested, it is unclear how the effect may be mediated. CREB binding protein (CBP) is a nuclear cofactor named for its ability to bind CREB. However, CBP also binds other nuclear proteins. We determined that CBP interacts with Pit-1 and is a cofactor for Pit-1-dependent activation of the human GH promoter. This pathway appears to be independent of CREB, with CPB being the likely target of phosphorylation by PKA.


Statins are ineffective at reducing neuroinflammation or prolonging survival in scrapie-infected mice.
Neuroinflammation is a prominent component of several neurodegenerative diseases, including multiple sclerosis, Alzheimer's disease, Parkinson's disease, tauopathies, amyotrophic lateral sclerosis and prion diseases. In such conditions, the ability to decrease neuroinflammation by drug therapy may influence disease progression. Statins have been used to treat hyperlipidemia as well as reduce neuroinflammation and oxidative stress in various tissues. In previous studies, treatment of scrapie-infected mice with the type 1 statins, simvastatin or pravastatin, showed a small beneficial effect on survival time. In the current study, to increase the effectiveness of statin therapy, we treated infected mice with atorvastatin, a type 2 statin that has improved pharmacokinetics over many type 1 statins. Treatments with either simvastatin or pravastatin were tested for comparison. We evaluated scrapie-infected mice for protease-resistant PrP (PrPres) accumulation, gliosis, neuroinflammation and time until advanced clinical disease requiring euthanasia. All three statin treatments reduced total serum cholesterol ≥40 % in mice. However, gliosis and PrPres deposition were similar in statin-treated and untreated infected mice. Time to euthanasia due to advanced clinical signs was not changed in statin-treated mice relative to untreated mice, a finding at odds with previous reports. Expression of 84 inflammatory genes involved in neuroinflammation was also quantitated. Seven genes were reduced by pravastatin, and one gene was reduced by atorvastatin. In contrast, simvastatin therapy did not reduce any of the tested genes, but did slightly increase the expression of Ccl2 and Cxcl13. Our studies indicate that none of the three statins tested were effective in reducing scrapie-induced neuroinflammation or neuropathogenesis.


Decision models for assessing the cost effectiveness of drug-eluting stents.
Percutaneous coronary intervention is an increasingly common treatment for many people with coronary disease. Randomised trials using antiproliferative, drug-eluting stents (DES) have shown important reductions in the need for repeat procedures after initial percutaneous coronary intervention. These trials indicate that the growing use of DES has the potential to create major clinical benefits for patients; however, in actual practice, the associated costs may cause financial crises for hospital systems. This paper reviews previously published DES economic decision models from both societal and hospital perspectives. From a social perspective, additional spending for new technologies may be acceptable if these technologies convincingly improve the lives of patients. The models reviewed suggest that DES incurs large costs to both society and hospitals, and currently offers uncertain clinical benefits. More research is recommended to identify high-risk patients who stand to gain significant health benefits from this therapy.


Oxia Planum: The Landing Site for the ExoMars "Rosalind Franklin" Rover Mission: Geological Context and Prelanding Interpretation.
The European Space Agency (ESA) and Roscosmos ExoMars mission will launch the "Rosalind Franklin" rover in 2022 for a landing on Mars in 2023.The goals of the mission are to search for signs of past and present life on Mars, investigate the water/geochemical environment as a function of depth in the shallow subsurface, and characterize the surface environment. To meet these scientific objectives while minimizing the risk for landing, a 5-year-long landing site selection process was conducted by ESA, during which eight candidate sites were down selected to one: Oxia Planum. Oxia Planum is a 200 km-wide low-relief terrain characterized by hydrous clay-bearing bedrock units located at the southwest margin of Arabia Terra. This region exhibits Noachian-aged terrains. We show in this study that the selected landing site has recorded at least two distinct aqueous environments, both of which occurred during the Noachian: (1) a first phase that led to the deposition and alteration of ∼100 m of layered clay-rich deposits and (2) a second phase of a fluviodeltaic system that postdates the widespread clay-rich layered unit. Rounded isolated buttes that overlie the clay-bearing unit may also be related to aqueous processes. Our study also details the formation of an unaltered mafic-rich dark resistant unit likely of Amazonian age that caps the other units and possibly originated from volcanism. Oxia Planum shows evidence for intense erosion from morphology (inverted features) and crater statistics. Due to these erosional processes, two types of Noachian sedimentary rocks are currently exposed. We also expect rocks at the surface to have been exposed to cosmic bombardment only recently, minimizing organic matter damage.


Using Social Constructivist Learning Theory to Unpack General Practitioners' Learning Preferences of End-of-Life Care: A Systematically Constructed Narrative Review.
General practitioners play a vital role in providing community-based palliative care to patients reaching end of life. In order for GPs to upgrade their skills at end-of-life care delivery, it is imperative that training programs be aligned to their learning needs and preferences. A narrative review was conducted using the electronic databases PubMed, CINAHL, PsycINFO, EMBASE, Scopus, Web of Science, and Cochrane from 01/01/1990 to 31/05/2021. 23 articles (of 10037 searched) were included for the review. Following themes were generated: Value attributed to end-of-life care learning, experience and reflection as a departure point for learning, learning as embedded in the clinical context; autonomy to decide upon their learning needs and learning preferences, learning as a transformative process; and learning as embedded in social interaction and interpretation. Training programs that are aligned to the preferences of GPs will encourage a larger clientele of GPs to access them.


Global patterns and controlling factors of soil nitrification rate.
Soil nitrification, an important pathway of nitrogen transformation in ecosystems, produces soil nitrate that influences net primary productivity, while the by-product of nitrification, nitrous oxide, is a significant greenhouse gas. Although there have been many studies addressing the microbiology, physiology, and impacting environment factors of soil nitrification at local scales, there are very few studies on soil nitrification rate over large scales. We conducted a global synthesis on the patterns and controlling factors of soil nitrification rate normalized at 25°C by compiling 3,140 observations from 186 published articles across terrestrial ecosystems. Soil nitrification rate tended to decrease with increasing latitude, especially in the Northern Hemisphere, and varied largely with ecosystem types. The soil nitrification rate significantly increased with mean annual temperature (MAT), soil nitrogen content, microbial biomass carbon and nitrogen, soil ammonium, and soil pH, but decreased with soil carbon:nitrogen and carbon:nitrogen of microbial biomass. The total soil nitrogen content contributed the most to the variations of global soil nitrification rate (total coefficient = 0.29) in structural equation models. The microbial biomass nitrogen (MBN; total coefficient = 0.19) was nearly of equivalent importance relative to MAT (total coefficient = 0.25) and soil pH (total coefficient = 0.24) in determining soil nitrification rate, while soil nitrogen and pH influenced soil nitrification via changing soil MBN. Moreover, the emission of soil nitrous oxide was positively related to soil nitrification rate at a global scale. This synthesis will advance our current understanding on the mechanisms underlying large-scale variations of soil nitrification and benefit the biogeochemical models in simulating global nitrogen cycling.


Accessory genetic content in Campylobacter jejuni ST21CC isolates from feces and blood.
Campylobacter jejuni is an important foodborne pathogen and the most commonly reported bacterial cause of gastroenteritis. C. jejuni is occasionally found in blood, although mechanisms important for invasiveness have remained unclear. C. jejuni is divided into many different lineages, of which the ST21 clonal complex (CC) is widely distributed. Here, we performed comparative genomic and in vitro analyses on 17C. jejuni ST21CC strains derived from human blood and feces in order to identify features associated with isolation site. The ST21CC lineage is divided into two large groups; centered around ST-21 and ST-50. Our clinical strains, typed as ST-50, showed further microevolution into two distinct clusters. These clusters were distinguished by major differences in their capsule loci and the distribution of accessory genetic content, including C. jejuni integrated elements (CJIEs) and plasmids. Accessory genetic content was more common among fecal than blood strains, whereas blood strains contained a hybrid capsule locus which partially consisted of C. jejuni subsp. doylei-like content. In vitro infection assays with human colon cell lines did not show significant differences in adherence and invasion between the blood and fecal strains. Our results showed that CJIEs and plasmid derived genetic material were less common among blood isolates than fecal isolates; in contrast, hybrid capsule loci, especially those containing C. jejuni subsp. doylei-like gene content, were found among many isolates derived from blood. The role of these findings requires more detailed investigation.


Divided-aperture technique for fluorescence confocal microscopy through scattering media.
We present a diffraction analysis for image formation in fluorescence confocal microscopy with divided apertures. The three-dimensional optical transform function is given, and axial resolution and transverse resolution are investigated. The results show that the employment of a divided-aperture technique in fluorescence confocal microscopy can enhance the rejection of background scattering. In addition, the axial resolution and transverse resolution can be improved by adjusting the width of the divider strip. For a given detector size, an optimum value of the divider strip width is given to obtain the best axial resolution or transverse resolution. The integrated intensity and signal-to-background ratio are also discussed.


Primary myelofibrosis: 2012 update on diagnosis, risk stratification, and management.
Primary myelofibrosis (PMF) is a myeloproliferative neoplasm characterized by stem cell-derived clonal myeloproliferation, abnormal cytokine expression, bone marrow fibrosis, anemia, splenomegaly, extramedullary hematopoiesis (EMH), constitutional symptoms, cachexia, leukemic progression, and shortened survival. Diagnosis is based on bone marrow morphology. The presence of fibrosis, JAK2/MPL mutation or +9/13q- cytogenetic abnormality is supportive but not essential for diagnosis. Prefibrotic PMF mimics essential thrombocythemia in its presentation and the distinction is prognostically relevant. Differential diagnosis of myelofibrosis should include chronic myelogenous leukemia, myelodysplastic syndromes, chronic myelomonocytic leukemia, and acute myeloid leukemia. The Dynamic International Prognostic Scoring System-plus (DIPSS-plus) prognostic model for PMF can be applied at any point during the disease course and uses eight independent predictors of inferior survival: age >65 years, hemoglobin <10 g/dL, leukocytes >25 × 10(9) /L, circulating blasts ≥1%, constitutional symptoms, red cell transfusion dependency, platelet count <100 × 10(9) /L, and unfavorable karyotype (i.e., complex karyotype or sole or two abnormalities that include +8, -7/7q-, i(17q), inv(3), -5/5q-, 12p- or 11q23 rearrangement). The presence of 0, 1, "2 or 3," and ≥4 adverse factors defines low, intermediate-1, intermediate-2, and high-risk disease with median survivals of ~15.4, 6.5, 2.9, and 1.3 years, respectively. A >80% two-year mortality is predicted by monosomal karyotype, inv(3)/i(17q) abnormalities, or any two of circulating blasts >9%, leukocytes ≥40 × 10(9) /L or other unfavorable karyotype. Observation alone is adequate for asymptomatic low/intermediate-1 risk disease. Allogeneic stem cell transplantation or experimental drug therapy is considered for intermediate-2/ high risk disease. Conventional or experimental drug therapy is reasonable for symptomatic intermediate-1 risk disease. Splenectomy and low-dose radiotherapy are used for drug-refractory splenomegaly. Radiotherapy is also used for the treatment of non-hepatosplenic EMH, PMF-associated pulmonary hypertension, and extremity bone pain.


Stable HIV-1 reservoirs on dolutegravir maintenance monotherapy: the MONODO study.
Dolutegravir (DTG) is a highly effective integrase inhibitor with a strong genetic resistance barrier and a potential role in simplified HIV maintenance treatment. We assessed the feasibility of DTG maintenance monotherapy and measured HIV reservoirs on DTG monotherapy. An interventional, open-label, single-arm study including eight virologically suppressed HIV-1-infected patients switched to DTG 50 mg once daily for 24 weeks was performed. HIV-1 RNA levels in plasma and cerebrospinal and seminal fluids were measured at baseline and week 24, as well as HIV-1 DNA in peripheral cells and DTG concentrations in these compartments. HIV-1 RNA remained undetectable in all samples of blood, cerebrospinal fluid and sperm throughout the 24 weeks, except for one cerebrospinal fluid sample with a value of 28 HIV-1 RNA copies/mL at week 24. One patient discontinued the study because of a neurological side effect. There was no change in the mean HIV-1 DNA level between baseline and week 24. Plasma and cerebrospinal fluid DTG concentrations reached therapeutic levels in all patients in these two compartments. In this small sample of carefully selected patients, HIV-1 reservoirs were well controlled on DTG monotherapy over a period of 24 weeks. Viral suppression was also maintained throughout follow-up.


Standardising visual control devices for tsetse flies: Central and West African species Glossina palpalis palpalis.
Glossina palpalis palpalis (G. p. palpalis) is one of the principal vectors of sleeping sickness and nagana in Africa with a geographical range stretching from Liberia in West Africa to Angola in Central Africa. It inhabits tropical rain forest but has also adapted to urban settlements. We set out to standardize a long-lasting, practical and cost-effective visually attractive device that would induce the strongest landing response by G. p. palpalis for future use as an insecticide-impregnated tool in area-wide population suppression of this fly across its range. Trials were conducted in wet and dry seasons in the Ivory Coast, Cameroon, the Democratic Republic of Congo and Angola to measure the performance of traps (biconical, monoconical and pyramidal) and targets of different sizes and colours, with and without chemical baits, at different population densities and under different environmental conditions. Adhesive film was used as a practical enumerator at these remote locations to compare landing efficiencies of devices. Independent of season and country, both phthalogen blue-black and blue-black-blue 1 m(2) targets covered with adhesive film proved to be as good as traps in phthalogen blue or turquoise blue for capturing G. p. palpalis. Trap efficiency varied (8-51%). There was no difference between the performance of blue-black and blue-black-blue 1 m(2) targets. Baiting with chemicals augmented the overall performance of targets relative to traps. Landings on smaller phthalogen blue-black 0.25 m(2) square targets were not significantly different from either 1 m(2) blue-black-blue or blue-black square targets. Three times more flies were captured per unit area on the smaller device. Blue-black 0.25 m(2) cloth targets show promise as simple cost effective devices for management of G. p. palpalis as they can be used for both control when impregnated with insecticide and for population sampling when covered with adhesive film.


The Microevolution and Epidemiology of Staphylococcus aureus Colonization during Atopic Eczema Disease Flare.
Staphylococcus aureus is an opportunistic pathogen and variable component of the human microbiota. A characteristic of atopic eczema (AE) is colonization by S. aureus, with exacerbations associated with an increased bacterial burden of the organism. Despite this, the origins and genetic diversity of S. aureus colonizing individual patients during AE disease flares is poorly understood. To examine the microevolution of S. aureus colonization, we deep sequenced S. aureus populations from nine children with moderate to severe AE and 18 non-atopic children asymptomatically carrying S. aureus nasally. Colonization by clonal S. aureus populations was observed in both AE patients and control participants, with all but one of the individuals carrying colonies belonging to a single sequence type. Phylogenetic analysis showed that disease flares were associated with the clonal expansion of the S. aureus population, occurring over a period of weeks to months. There was a significant difference in the genetic backgrounds of S. aureus colonizing AE cases versus controls (Fisher exact test, P = 0.03). Examination of intra-host genetic heterogeneity of the colonizing S. aureus populations identified evidence of within-host selection in the AE patients, with AE variants being potentially selectively advantageous for intracellular persistence and treatment resistance.


Network instability dynamics drive a transient bursting period in the developing hippocampus in vivo.
Spontaneous correlated activity is a universal hallmark of immature neural circuits. However, the cellular dynamics and intrinsic mechanisms underlying network burstiness in the intact developing brain are largely unknown. Here, we use two-photon Ca2+ imaging to comprehensively map the developmental trajectories of spontaneous network activity in the hippocampal area CA1 of mice in vivo. We unexpectedly find that network burstiness peaks after the developmental emergence of effective synaptic inhibition in the second postnatal week. We demonstrate that the enhanced network burstiness reflects an increased functional coupling of individual neurons to local population activity. However, pairwise neuronal correlations are low, and network bursts (NBs) recruit CA1 pyramidal cells in a virtually random manner. Using a dynamic systems modeling approach, we reconcile these experimental findings and identify network bi-stability as a potential regime underlying network burstiness at this age. Our analyses reveal an important role of synaptic input characteristics and network instability dynamics for NB generation. Collectively, our data suggest a mechanism, whereby developing CA1 performs extensive input-discrimination learning prior to the onset of environmental exploration.


Response to beta-blockers and natriuretic peptide level in acute heart failure: analysis of data from the Korean acute heart failure registry.
To investigate the effect of beta-blockers according to NP levels and HF phenotypes because natriuretic peptide (NP) level can be used to risk-stratify HF patients regardless of left ventricular ejection fraction (LVEF). Of 5,625 patients in the Korean acute heart failure registry, we included patients with LVEF and NP levels. HF phenotypes were defined as HF with reduced ejection fraction (HFrEF) (EF ≤ 40%), HF with midrange ejection fraction (HFmrEF) (40% < EF < 50%), and HF with preserved EF (HFpEF) (EF ≥ 50%). Patients were further stratified by NP tertiles. Primary outcome was 5-year all-cause mortality according to beta-blocker use at discharge. Both B-type NP (BNP) (r = -0.279, P < 0.001) and N-terminal pro-BNP (r = -0.186, P < 0.001) levels correlated inversely with LVEF. During a median follow-up duration of 961 days, 1560 (35.3%) patients died. In HFrEF, patients taking beta-blockers showed better survival regardless of NP levels. Regarding HFmrEF, there was no mortality difference between those taking and not taking beta-blockers. In HFpEF, beta-blocker use demonstrated lower mortality in those in the 3rd NP tertile (log-rank P = 0.041) but not in those in the 1st and 2nd NP tertiles (log-rank P > 0.05). After adjusting covariates, the use of beta-blockers was associated with a 38%-reduced mortality (hazard ratio: 0.62; 95% confidence interval: 0.39-0.98; P = 0.040) in HFpEF patients in the 3rd NP tertile but not in those in 1st and 2nd tertiles. We confirm that the use of beta-blockers is beneficial in patients with HFrEF. Furthermore, we extend the benefits of beta-blockers to patients with HFpEF and high NP levels. ClinicalTrial.gov identifier: NCT01389843 URL: https://clinicaltrials.gov/ct2/show/NCT01389843.


Does sexual abuse in childhood cause pedophilia: an exploratory study.
The reliability of the notion that pedophilia is caused by sexual abuse in childhood was explored by examining retrospective self-reports of 344 males. Included in the study were 77 heterosexual pedophiles, 54 homosexual pedophiles, 51 nonpedophilic sex offenders against children, 36 sex offenders against physically mature females, 75 heterosexual paid volunteers who erotically preferred mature females, and 51 homosexual clients who preferred mature males. For each sex offender the differential diagnosis of an erotic preference for minors vs. a preference for physically mature partners was made by means of the phallometric test of erotic gender and age preferences. The analysis of self-reports confirmed that the proportion of pedophiles who report having been sexually abused in childhood by mature persons is larger than that of men who were not charged for or accused of a sex offense against a child though the difference is relatively small (28.6 vs. 13.9 and 10.7% for the heterosexual pedophiles and the two groups of gynephiles, respectively, and 25.9 vs. 11.8% for the homosexual pedophiles and androphiles, respectively). Further analysis demonstrated, however, that pedophiles who admitted having an erotic interest in children significantly more often claimed that they had been sexually abused as children than pedophiles who did not admit having such feelings. This interdependence renders the reliability of these self-reports questionable.


Lethal Sandslides from Eolian Dunes.
Fossil vertebrates entombed within the Upper Cretaceous Djadokhta Formation of southern Mongolia bear testimony to a heretofore unknown geologic phenomenon: mass wasting of eolian dunes during heavy rainstorms. Evaporation of shallow-penetrating rainwater led to progressive calcite accumulation in a thin layer of sand about 0.5 m below the surface of dune lee slopes. During rare heavy rainstorms, a perched water table developed at the top of calcitic zones. Positive pore water pressure led to translational slides and fast-moving sediment gravity flows that overwhelmed animals on the lee slopes of large dunes and in interdune areas.


Successful pregnancy in one horn of a bicornuate uterus.
Normal development of the female reproductive tract involves a series of complex processes characterized by the differentiation, migration, fusion, and subsequent canalization of the Müllerian system. Uterine anomalies result when these processes are interrupted. Bicornuate uterus is a unification defect of the Müllerian ducts, and it is estimated to represent 10-39% of Müllerian duct anomalies. Pregnancies in bicornuate uterus are usually considered high-risk because of association with poor reproductive outcomes, such as pregnancy loss, preterm birth, malpresentations, and fetal deformity. Routine cervical cerclage and Strassman metroplasty have been advocated as treatment for this anomaly. We report a case of successful pregnancy in one of the horns of a bicornuate uterus managed in our unit.


Vaccine safety studies of Brucella abortus S19 and S19ΔvjbR in pregnant swine.
Brucellosis in swine is caused by Brucella suis, a bacterial infection of nearly worldwide distribution. Brucella suis is also transmissible to humans, dogs and cattle and is considered a reemerging disease of public health concern. To date, there is no effective vaccine for swine. This prompted us to investigate the potential use of the commercially available vaccine for cattle or the live attenuated vaccine candidate S19ΔvjbR. As the first step, we sought to study the safety of the vaccine candidates when administered in pregnant sows, since one of the major drawbacks associated with vaccination using Live Attenuated Vaccines (LAV) is the induction of abortions when administered in pregnant animals. Fifteen pregnant gilts at mid-gestation were divided into four groups and subsequently vaccinated subcutaneously using different formulations containing 2.0 ± 0.508 × 109 CFU of either S19 or S19ΔvjbR. Vaccination in pregnant animals with the vaccine candidates did not induce abortion, stillbirths or a reduction in litter size. Multiple tissues in the gilts and piglets were examined at the time of delivery to assess bacterial colonization and histopathological changes. There was no evidence of vaccine persistence in the gilts or bacterial colonization in the fetuses. Altogether, these data suggest that both vaccine candidates are safe for use in pregnant swine. Analysis of the humoral responses, specifically anti-Brucella IgG levels measured in serum, demonstrated a robust response induced by either vaccine, but of shorter duration (4-6 weeks post-inoculation) compared to that observed in cattle or experimentally infected mice. Such a transient humoral response may prove to be beneficial in cases where the vaccine is used in eradication campaigns and in the differentiation of vaccinated from infected animals. This study provides evidence to support future efficacy studies of both vaccine candidates in swine.


Upper extremity venous thrombosis in hospitalized patients: A prospective epidemiological study.
The frequency of upper extremities vein thrombosis (UEVT) is rising with the increasing use of endovenous devices. These thromboses are particularly common among hospitalized patients. The epidemiology and risk factors for UEVT are poorly understood in a hospitalized population. To assess the prevalence of UEVT in hospitalized patients and study thrombosis risk factors according to their location. Prospective evaluation of patients hospitalized in a university hospital with clinical and Doppler ultrasound (DUS) assessment of the upper extremities. Of the 400 patients included, 91(22.8 %) had UEVT including 8 (8.8 %) proximal thrombosis, 32 (35.2 %) arm venous thrombosis and 51 (56.0 %) forearm venous thrombosis; 7 (7.7 %) patients with UEVT had a concomitant symptomatic pulmonary embolism. In this population of hospitalized patients, 40 (10 %) had proximal or arm thrombosis and 51 (12.8 %) forearm thrombosis. All patients with UEVT had a venous catheter and 63 (69.2 %) of UEVT patients had therapeutic or prophylactic anticoagulation at the time of DUS evaluation. In multivariate analysis, peripheral intravenous catheter (PIVC) OR 3.71 [1.90; 7.91] (p < 0.001); MID line OR 3.58 [1.46; 8.91] (p = 0.005) and infection disease OR 2.21 [1.26; 4386] (p = 0.005) were associated with UEVT. Central venous catheter OR 66.24 [12.64; 587.03] was associated with proximal UEVT and MID line OR 12.61 [4.64; 35.77] (p < 0.001) with arm UEVT. Forearm UEVT were associated with PIVCOR 10.71[3.14; 67.37] (p = 0.001); infectious disease OR 2.87 [1.48; 5.60] (p = 0.002), iron infusion OR 3.11 [1.23; 7.47] and hospitalization for postpartum OR 4.37 [0.97; 18.06] (p = 0.04). One in 5 hospitalized patients suffers from UEVT. Proximal venous thrombosis is uncommon. The presence of a catheter and hospitalization for infection are the factors most frequently associated with UEVT.


A guide to GRADE guidelines for the readers of JTH.
More than 70 organizations worldwide have adopted the GRADE methodology for guideline development. The ninth iteration of the American Collage of Chest Physicians guidelines (AT9) adopted structural and policy changes that resulted in a greater adherence to GRADE guidance than previous iterations. The most important of these changes include minimizing the impact of financial and intellectual conflict of interest, increasing the rigor of evidence evaluation, acknowledging uncertainty in estimates of typical values and preferences, and awareness of the large variability in values and preferences. One of the consequences of the greater adherence to GRADE methodology is an increase in weak vs. strong recommendations in AT9. The result of the GRADE process highlights the desirability of higher-quality evidence both regarding the outcomes of alternative management strategies and regarding the distribution of values and preferences in patients considering those alternatives. It also encourages shared decision making in encounters between physicians and patients. Although some physicians might find the uncertainty underlying medical practice discouraging or unsettling, relative to denying or obscuring the uncertainty, acknowledging and addressing the uncertainty will lead to more credible, realistic, and useful recommendations.


Trends in motorcycle fatalities associated with alcohol-impaired driving--United States, 1983-2003.
Motorcycles are the most dangerous type of motor vehicle to drive. These vehicles are involved in fatal crashes at a rate of 35.0 per 100 million miles of travel, compared with a rate of 1.7 per 100 million miles of travel for passenger cars. The National Highway Traffic Safety Administration (NHTSA) has reported increasing numbers of motorcycle deaths associated with alcohol-impaired driving in recent years, especially among persons aged > or =40 years. To determine trends by age group in motorcycle fatalities overall and in those involving alcohol impairment, CDC analyzed data from the NHTSA Fatality Analysis Reporting System (FARS) for 1983, 1993, and 2003. This report summarizes the results of that analysis, which indicated that, during 1983-2003, the overall prevalence of elevated blood alcohol concentrations (BACs) among motorcycle drivers who died in crashes declined; however, the peak rate of death among alcohol-impaired motorcycle drivers shifted from those aged 20-24 years to those aged 40-44 years. Strong enforcement of existing BAC laws, together with other public health interventions aimed at motorcyclists, might reduce the crash mortality rate, especially among older drivers.


Evaluation of a droplet digital PCR assay for quantification of Mycobacterium avium subsp. paratuberculosis DNA in whole-blood and fecal samples from MAP-infected Holstein cattle.
Bovine paratuberculosis (PTB) is an infectious disease that affects ruminants worldwide and is a burden on the dairy industry. PTB control measures include culling of Mycobacterium avium subsp. paratuberculosis (MAP)-infected animals from the herd and the enhancement of farm-biosecurity measures. Diagnostics tools for the direct detection of MAP are fecal real-time qPCR and bacteriological culture, the last one being considered the gold standard. However, both show limitations for detecting subclinical MAP-infected cattle with low bacterial load in feces and gut tissues. Droplet digital polymerase chain reaction (ddPCR) is a third-generation PCR method that shows high reproducibility for the quantification of low DNA copy numbers. The objective of this study was to design a ddPCR assay to detect and quantify a fragment of the F57 MAP-specific sequence in samples of naturally MAP-infected Holstein cattle. DNA was isolated from whole-blood and fecal samples from control cows with a negative ELISA and qPCR result (N = 75) and from cows with PTB-associated focal (N = 32), multifocal (N = 21), and diffuse lesions (N = 17) in gut tissues. After ddPCR, the DNA extracted from fecal samples of cows with diffuse lesions showed higher mean copies per microliter (13,791.2 copies/μl) than samples from cows with multifocal lesions (78.8 copies/μl), focal lesions (177.1 copies/μl) or control cows (4.8 copies/μl) (P ≤ 0.05). Significant differences in mean DNA copies/μl were also observed in the blood samples from cows with focal lesions (47.7 copies/μl) when compared with cows with multifocal and diffuse lesions; 18.1 and 12.4 copies/μl, respectively. Using a principal component analysis, the results of the fecal ddPCR clustered together with the results of a commercial ELISA for the specific detection of MAP antibodies, fecal and tissue qPCR, and bacteriological culture results. In contrast, blood ddPCR results clustered together with the results of an ELISA for the detection of a biomarker of subclinical PTB, the ABCA13 transporter. Blood ddPCR was the most sensitive tool (sensitivity 71%, specificity 100%) of all the quantitative methods used in the study for the detection of subclinical cows with focal lesions.


Yeast DNA polymerase--DNA primase complex; cloning of PRI 1, a single essential gene related to DNA primase activity.
The immunopurified yeast DNA polymerase--DNA primase complex is constituted by DNA polymerase I polypeptides and by three other protein species, called p74, p58 and p48, which we show to be immunologically unrelated. The gene encoding the p48 polypeptide has been identified by immunological screening of a lambda gt11 yeast genomic DNA library. Antiserum specific for p48 inhibits DNA primase, and immunoreactive, inhibitory antibodies are affinity-purified by the clone-encoded protein, thus relating the p48 polypeptide to DNA primase activity. The entire gene has been cloned, and the 1.45-kb p48 mRNA is overproduced in cells containing the gene in high copy number. Gene disruption and Southern hybridization experiments demonstrate that the p48 protein is encoded by a single gene and it performs an essential function.


The relationship between microstructure and in vivo degradation of modified bacterial cellulose sponges.
Bacterial cellulose (BC) and hydroxyapatite (HA) possess unique structures and excellent biocompatibility. Considerable work has been performed to develop composites that promote bone repair. However, the use of BC/HA composites is limited because the lack of corresponding enzymes makes them non-degradable in vivo. In the present study, C6-carboxylated bacterial cellulose (TBC) was prepared in a bromide-free system. Several composite methods of TBC and HA are compared, including in situ formation, physical mixing and biomineralization. Composite sponges prepared by different methods were characterized by tensile testing, X-ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscopy and in vivo degradation. The structural anisotropy of various sponges was analyzed to quantitatively evaluate their microstructure. The results suggest that the interaction between HA and TBC nanofibers has a large influence on microstructure and macroscopic properties. Moreover, the structural anisotropy and the speed of granulation ingrowth were strongly interdependent. This improved understanding of slowly degrading BC-based materials suggests that modified cellulose-based materials can be made degradable by altering their microstructure.


Expression hierarchy in the Yersinia type III secretion system established through YopD recognition of RNA.
The Yersinia type III secretion system (T3SS) is environmentally responsive to enable its rapid induction upon contact with host cells and is necessary for Yersiniae to establish a replicative niche and cause disease. YopD, a translocator protein, represses the expression of T3SS genes until signalled by environmental cues, a mechanism known as the low calcium response. In this work, we investigated recognition of target genes by Yersinia pestis YopD. Expression of all genes of the T3SS was induced in a yopD mutant, though not to the same degree, with effector Yops most affected. Two, short AU-rich sequence elements up- and downstream of start codons of target genes were necessary but not sufficient for YopD mediated repression. Purified YopD-LcrH bound specifically to target RNAs in vitro with different relative affinities, with effector Yops having greater affinity. Together, the data suggest YopD binds to T3SS transcripts where it may prevent ribosome binding causing accelerated mRNA degradation. This regulatory mechanism may ensure an expression hierarchy during the low calcium response as low affinity YopD targets such as chaperones would be translated prior to high affinity targets such as effector Yops allowing the bacteria another layer of control over Yop translocation during infection.


[Gaucher's disease type I: an infrequent cause of portal hypertension].
Portal hypertension associated to Gaucher's disease type I with non cirrhotic liver involvement is a very in-equent finding. We report a case of Gaucher's disease type I without cirrhosis in which portal hypertension is the main manifestation. The mechanism of the portal hypertension is discussed in our report and can be related to an elevation in hepatic fluid resistance due to the Gaucher's cells compression on sinusoids.


The evaluation of 1,4-benzoquinones as inhibitors of human monoamine oxidase.
The monoamine oxidase (MAO) enzymes are of considerable pharmacological interest and inhibitors are used in the clinic for the treatment of major depressive disorder and Parkinson's disease. A limited number of studies have shown that the quinone class of compounds possesses MAO inhibition properties. Most notable among these is a report that 2,3,6-trimethyl-1,4-naphthoquinone (TMN), present in extracts of cured tobacco leafs, is a non-selective inhibitor of both MAO isoforms. An older study reports that 1,4-benzoquinone inhibits MAO-A and MAO-B from human synaptosomes. Both 1,4-naphthoquinones and 1,4-benzoquinone are reported to inhibit the MAOs with a reversible mode of action. Since the MAO inhibition properties of additional members of the 1,4-benzoquinone class of compounds have not yet been explored, the present study investigates a small series of four 1,4-benzoquinones which incorporate phenyl, benzyl, benzyloxy and cyclopentyl monosubstitution on C2. The 1,4-benzoquinones were found to be moderately potent MAO inhibitors with IC50 values of 5.03-13.2 μM (MAO-A) and 3.69-23.2 μM (MAO-B). These values are comparable to those recorded for 1,4-benzoquinone of 4.82 μM (MAO-A) and 10.2 μM (MAO-B). Of interest however, is the finding that the 1,4-benzoquinones are irreversible inhibitors of MAO-A since prolonged incubation results in near complete inhibition, and enzyme activity is not recovered by dialysis. MAO-B is much less sensitive to inactivation by the 1,4-benzoquinones. These findings are discussed with reference to a possible mechanism by which irreversible inhibition occurs. It may be concluded that irreversible 1,4-benzoquinone-derived inhibitors may act as probes for investigating quinone reactive sites in the MAOs.


Highly Selective p-Xylene Separation from Mixtures of C8 Aromatics by a Nonporous Molecular Apohost.
High and increasing production of separation of C8 aromatic isomers demands the development of purification methods that are efficient, scalable, and inexpensive, especially for p-xylene, PX, the largest volume C8 commodity. Herein, we report that 4-(1H-1,2,4-triazol-1-yl)-phenyl-1H-benzo[de]isoquinoline-1,3(2H)-dione (TPBD), a molecular compound that can be prepared and scaled up via solid-state synthesis, exhibits exceptional PX selectivity over each of the other C8 isomers, o-xylene (OX), m-xylene (MX), and ethylbenzene (EB). The apohost or α form of TPBD was found to exhibit conformational polymorphism in the solid state enabled by rotation of its triazole and benzene rings. TPBD-αI and TPBD-αII are nonporous polymorphs that transformed to the same PX inclusion compound, TPBD-PX, upon contact with liquid PX. TPBD enabled highly selective capture of PX, as established by competitive slurry experiments involving various molar ratios in binary, ternary, and quaternary mixtures of C8 aromatics. Binary selectivity values for PX as determined by 1H NMR spectroscopy and gas chromatography ranged from 22.4 to 108.4, setting new benchmarks for both PX/MX (70.3) and PX/EB (59.9) selectivity as well as close to benchmark selectivity for PX/OX (108.4). To our knowledge, TPBD is the first material of any class to exhibit such high across-the-board PX selectivity from quaternary mixtures of C8 aromatics under ambient conditions. Crystallographic and computational studies provide structural insight into the PX binding site in TPBD-PX, whereas thermal stability and capture kinetics were determined by variable-temperature powder X-ray diffraction and slurry tests, respectively. That TPBD offers benchmark PX selectivity and facile recyclability makes it a prototypal molecular compound for PX purification or capture under ambient conditions.


Reproducible high yield of rat islets by stationary in vitro digestion following pancreatic ductal or portal venous collagenase injection.
Pancreatic distension with collagenase solution followed by stationary in vitro digestion yields large numbers of intact islets. We compared in rats two routes of collagenase injection, pancreatic ductal (PD) and portal venous (PV), for islet yield, in vitro insulin secretory capacities, and in vivo functional viability. The islet yield in the PD method (n = 11) was greater than that in the PV method (n = 8) (682 +/- 27 vs. 417 +/- 39 per pancreas, P less than 0.025). The insulin release from the PD islets in response to 16.7 mM glucose increased gradually following culture, 3.2 +/- 0.8 ng/10 islets/30 min (fresh) to 12.3 +/- 2.1 (24-hr culture). In contrast, insulin release from the PV islets increased during the first 6 hr of culture, but decreased after 24 hr in culture. Under electronmicroscopic examination, the PD islets revealed a well preserved structure with healthy endocrine cells, while the PV islets showed a dilated capillary network and distorted endocrine cell continuity. When 100 PD islets were transplanted into streptozotocin-induced diabetic B6AF1 mice (n = 8), all the recipient mice restored normoglycemia (less than 200 mg/dl) within 1-4 days following transplantation and maintained it until rejection. However, the recipient mice given 100 PV islets showed a significant delay in restoring normoglycemia, and 3 of 8 mice given 100 PV islets were still hyperglycemic on day 4 postgrafting. In summary, pancreatic ductal collagenase injection followed by stationary in vitro digestion reproducibly yields higher numbers of intact and viable islets when compared with portal venous collagenase injection, indicating the superiority of this method to portal venous injection.


Nonnegative principal component analysis for mass spectral serum profiles and biomarker discovery.
As a novel cancer diagnostic paradigm, mass spectroscopic serum proteomic pattern diagnostics was reported superior to the conventional serologic cancer biomarkers. However, its clinical use is not fully validated yet. An important factor to prevent this young technology to become a mainstream cancer diagnostic paradigm is that robustly identifying cancer molecular patterns from high-dimensional protein expression data is still a challenge in machine learning and oncology research. As a well-established dimension reduction technique, PCA is widely integrated in pattern recognition analysis to discover cancer molecular patterns. However, its global feature selection mechanism prevents it from capturing local features. This may lead to difficulty in achieving high-performance proteomic pattern discovery, because only features interpreting global data behavior are used to train a learning machine. In this study, we develop a nonnegative principal component analysis algorithm and present a nonnegative principal component analysis based support vector machine algorithm with sparse coding to conduct a high-performance proteomic pattern classification. Moreover, we also propose a nonnegative principal component analysis based filter-wrapper biomarker capturing algorithm for mass spectral serum profiles. We demonstrate the superiority of the proposed algorithm by comparison with six peer algorithms on four benchmark datasets. Moreover, we illustrate that nonnegative principal component analysis can be effectively used to capture meaningful biomarkers. Our analysis suggests that nonnegative principal component analysis effectively conduct local feature selection for mass spectral profiles and contribute to improving sensitivities and specificities in the following classification, and meaningful biomarker discovery.