Comprehensive analysis of miRNA-mRNA/lncRNA during gonadal development of triploid female rainbow trout (Oncorhynchus mykiss).
Chromosomal ploidy manipulation is one of the means to create excellent germplasm. Triploid fish could provide an ideal sterile model for searching of a underlying mechanism of abnormality in meiosis. The complete understanding of the coding and noncoding RNAs regulating sterility caused by meiosis abnormality is still not well understood. By high-throughput sequencing, we compared the expression profiles of gonadal mRNA, long non-coding RNA (lncRNA), and microRNA (miRNA) at three different developmental stages between the diploid (XX) and triploid (XXX) female rainbow trout. These stages were gonads before differentiation (65 days post fertilisation, dpf), at the beginning of morphological differences (180 dpf) and showing clear difference between diploids and triploids (600 dpf), respectively. A majority of differentially expressed (DE) RNAs were identified, and 22 DE mRNAs related to oocyte meiosis and homologous recombination were characterized. The predicted miRNA-mRNA/lncRNA networks of 3 developmental stages were constructed based on the target pairs of DE lncRNA-miRNA and DE mRNA-miRNA. According to the networks, meiosis-related gene of ccne1 was targeted by dre-miR-15a-5p_R + 1, and 6 targeted DE lncRNAs were identified. Also, qRT-PCR was performed to validate the credibility of the network. Overall, this study explored the potential interplay between coding and noncoding RNAs during the gonadal development of polyploid fish. The mRNA, lncRNA and miRNA screened in this study may be helpful to identify the functional elements regulating fertility of rainbow trout, which may provide reference for character improvement in aquaculture.


Impact of Anterior Kidney Calyx Involvement of Complex Stones on Outcomes for Patients Undergoing Percutaneous Nephrolithotomy.
A number of factors that can impact the outcomes of percutaneous nephrolithotomy (PCNL) procedures have previously been investigated. Complex stones that extend to the anterior calyx could affect the success and complication rates of PCNL. We analyzed the effect of anterior calyx involvement on the outcomes of patients with complex stones treated with PCNL. A total of 132 consecutive patients who underwent PCNL due to complex stones (multiple, partial staghorn, or staghorn stones) between 2015 and 2017 were enrolled in this study. They were stratified into two groups based on whether the stone extended to the anterior calyx (group 1, n = 45) or not (group 2, n = 87). The stratification was achieved through contrast-enhanced computerized tomography (CT). Demographics, laboratory tests, and peri- and postoperative findings (operation and fluoroscopy duration, hospital stay, utilization of flexible instruments, access numbers, total blood count change, stone-free rate [SFR], and complications) were compared between the groups. The SFR was evaluated by plain kidney-ureter-bladder radiography or CT. The demographics, operation and fluoroscopy duration, access number, and hospital stay were similar between the groups (p < 0.05). A higher drop in the hemoglobin level in group 1 was identified (group 1 [2.14 ± 1.49 g/dL] vs. group 2 [1.43 ± 1.31 g/dL]) (p = 0.006). The SFR among the patients with extension to the anterior calyx was 60%, compared to 77% among the patients with no extension to the anterior calyx (p = 0.041). Flexible instruments were utilized in 60% of the patients of group 1, which was a higher rate than for group 2 (36%) (p = 0.007). Complication rates were similar in the two groups according to the Clavien-Dindo classification (p > 0.05). Our study demonstrated that complicated stones with extension to the anterior calyx are more challenging than cases without extension to the anterior calyx. This was noted by a lower SFR, a more prominent drop in total blood count, and more frequent utilization of flexible scopes.


Anti-beta2-glycoprotein I antibodies as risk factors for acute myocardial infarction.
To determine whether high levels of antibodies against the phospholipid beta2-glycoprotein I (beta2-gpI) cofactor are associated with an increase in the risk of acute myocardial infarction. The study comprised 82 patients with acute myocardial infarction and 82 controls, who were assessed in regard to age, sex, race, hypertension, smoking, previous heart disease, history of diabetes mellitus, and hypercholesterolemia. The following antibodies were detected using immunoassay: anticardiolipin and anti-beta2-gpI IgA, IgG, and IgM. Adjusted odds ratios (OR) for risk factors were obtained through logistic regression. The mean ages of the cases and controls were, respectively, 57.7 and 51.1 years (P=0.003). Men (P=0.005) and the white race predominated in both groups (P=0.798). Of the risk factors, a history of diabetes (OR=5.3; 95% CI: 1.9 to 14.9; P=0.001) and previous heart disease (OR=4.7; 95% CI: 2.0 to 10.7; P<0.001) were the most consistent associations with myocardial infarction. The frequency of anticardiolipin IgG, IgM, and IgA antibodies did not differ between cases and controls (P=1.000). Anti-beta2-gpI IgA antibodies were more frequent in cases than in controls (P=0.054). The adjusted OR for anti-beta2-gpI IgA antibodies was 3.4 (95% CI: 1.3 to 9.1; P=0.015). Anti-beta2-gpI IgA antibodies, but not anticardiolipin antibodies, seemed to behave as independent risk factors for myocardial infarction, which may represent a link between autoimmunity and atherosclerosis in patients with acute myocardial infarction.


B virus, Herpesvirus simiae: historical perspective.
Between 1932 and 1972, 24 known infections of man by B virus caused 23 cases of encephalitis and 18 fatalities. The virus has been isolated from dermal lesions and neural ganglia from macaque monkeys. Serological evidence of infection is complicated by close antigenic relationships between B virus, Herpesvirus simplex, and SA8. Hyperimmune globulin produced from monkey, horse, and rabbit sera has not proved highly effective. Formalin-inactivated vaccine appears safe and antigenic in man but has not been licensed. Half of all human subjects have neutralizing B virus antibody related to their H. simplex titer. More stable animal populations and the improved use of protective apparel have reduced, but not eliminated, the risk of B virus to man.


Reactive oxygen species promote ovarian cancer progression via the HIF-1α/LOX/E-cadherin pathway.
Reactive oxygen species (ROS) can drive the de‑differentiation of tumor cells leading to the process of epithelial-to-mesenchymal transition (EMT) to enhance invasion and metastasis. The invasive and metastatic phenotype of malignant cells is often linked to loss of E-cadherin expression, a hallmark of EMT. Recent studies have demonstrated that hypoxic exposure causes HIF-1-dependent repression of E-cadherin. However, the mechanism by which ROS and/or HIF suppresses E-cadherin expression remains less clear. In the present study, we found that ROS accumulation in ovarian carcinoma cells upregulated HIF-1α expression and subsequent transcriptional induction of lysyl oxidase (LOX) which repressed E-cadherin. Loss of E-cadherin facilitated ovarian cancer (OC) cell migration in vitro and promoted tumor growth in vivo. E-cadherin immunoreactivity correlated with International Federation of Gynecology and Obstetrics (FIGO) stage, tumor differentiation and metastasis. Negative E-cadherin expression along with FIGO stage, tumor differentiation and metastasis significantly predicted for a lower 5-year survival rate. These findings suggest that ROS play an important role in the initiation of metastatic growth of OC cells and support a molecular pathway from ROS to aggressive transformation which involves upregulation of HIF-1α and its downstream target LOX to suppress E-cadherin expression leading to an increase in cell motility and invasiveness.


Multiple Participants' Discrete Activity Recognition in a Well-Controlled Environment Using Universal Software Radio Peripheral Wireless Sensing.
Wireless sensing is the utmost cutting-edge way of monitoring different health-related activities and, concurrently, preserving most of the privacy of individuals. To meet future needs, multi-subject activity monitoring is in demand, whether it is for smart care centres or homes. In this paper, a smart monitoring system for different human activities is proposed based on radio-frequency sensing integrated with ensemble machine learning models. The ensemble technique can recognise a wide range of activity based on alterations in the wireless signal's Channel State Information (CSI). The proposed system operates at 3.75 GHz, and up to four subjects participated in the experimental study in order to acquire data on sixteen distinct daily living activities: sitting, standing, and walking. The proposed methodology merges subject count and performed activities, resulting in occupancy count and activity performed being recognised at the same time. To capture alterations owing to concurrent multi-subject motions, the CSI amplitudes collected from 51 subcarriers of the wireless signals were processed and merged. To distinguish multi-subject activity, a machine learning model based on an ensemble learning technique was designed and trained using the acquired CSI data. For maximum activity classes, the proposed approach attained a high average accuracy of up to 98%. The presented system has the ability to fulfil prospective health activity monitoring demands and is a viable solution towards well-being tracking.


[Hyperuricemic variant of latent glomerulonephritis].
The authors describe the data of a comparative clinicomorphological analysis made in 40 patients with associated latent nephritis and hyperuricemia and 43 patients suffering from latent nephritis without hyperuricemia. No substantial differences in the disease clinical picture or in the structure of the morphological variants of glomerulonephritis (GN) were found in the above groups of patients. A more detailed morphological study made it possible to reveal a higher incidence of tubulostromal-vascular changes in patients with hyperuricemia. It is assumed that excess uric acid is likely to play an etiological role in the development of latent GN. Emphasis is laid on the necessity of early diagnosis of purine metabolism disorders in patients with latent nephritis whose goal-oriented therapy may improve the characteristics of the uric acid syndrome and renal function.


Reaction kinetics at very low temperatures: laboratory studies and interstellar chemistry.
Studies of gas-phase processes at temperatures down to 10 K have recently blossomed, largely through application of the CRESU (cinétique de réaction en ecoulement supersonique uniforme) technique. The results are of considerable relevance to the synthesis of molecules in dense interstellar clouds, demonstrating that the models developed to explain the observed molecular abundances must be expanded to include reactions between electrically neutral species. In addition, the experimental results have stimulated theoretical efforts to describe the factors that control the rates of such low-temperature reactions. In this Account, the CRESU method is described and the relevance of the results discussed.


The possible role of maternal bonding style and CHRNB2 gene polymorphisms in nicotine dependence and related depressive phenotype.
Neuronal nicotinic acetylcholinergic receptors (nAChR) and especially α4β2 nAChRs are the major targets for cessation medications and also for some promising antidepressant agents. Furthermore, depressive symptoms pose multifacet difficulties during cessation therapy. However, gene encoding for the β2 subunit of nAChRs has been poorly investigated in association with depression. Since both nicotine dependence (ND) and depressive phenotype are complex disorders, we investigated the effects of a significant early life experience, maternal bonding style (MB) and CHRNB2 gene SNPs on smoking-related depression. We recruited two hundred and thirty-two treatment-seeking smokers in our study. Phenotypic variants were evaluated using the Fagerstrom Test for Nicotine Dependence (FTND), the Zung Self-Rating Depression Scale (ZSDS) and the Parental Bonding Instrument (PBI). Besides the total score (TS) of ZSDS, impulsivity (ZSDS-I) and suicidal ideation (ZSDS-S) were distinguished as phenotypic variable. DNAs were extracted from buccal mucosa samples and one SNP in promoter and two SNPs in 3' UTR of CHRNB2 gene were genotyped. GLM and ANOVA tests were performed for genotype associations and interaction analyses. Maternal bonding had a significant impact on depressive phenotypes. Low care, high protection and affectionless control (ALC) were associated with ZSDS-TS and all subphenotypes of ZSDS. One SNP, the rs2072660 in 3' UTR, had a significant effect on the FTND score (p=0.010). Direct association of CHRNB2 variants and depressive phenotypes were not significant. However, in interaction with ALC, rs2072660 was significantly associated with ZSDS-S (p=0.005). MB had no significant effect on smoking-related phenotype. Our results highlight the important role of 3' UTR in the CHRNB2 gene in the shared molecular background of ND and depressive phenotype. Parental bonding style can be suggested as a significant environmental factor in further GxE studies of depression. The presented significant GxE interaction on smoking-related suicidal subphenotype may help establish further investigations on development of more effective and safer smoking cessation and antidepressant agents.


Leptin rapidly suppresses insulin release from insulinoma cells, rat and human islets and, in vivo, in mice.
Obesity is associated with diabetes, and leptin is known to be elevated in obesity. To investigate whether leptin has a direct effect on insulin secretion, isolated rat and human islets and cultured insulinoma cells were studied. In all cases, mouse leptin inhibited insulin secretion at concentrations within the plasma range reported in humans. Insulin mRNA expression was also suppressed in the cultured cells and rat islets. The long form of the leptin receptor (OB-Rb) mRNA was present in the islets and insulinoma cell lines. To determine the significance of these findings in vivo, normal fed mice were injected with two doses of leptin. A significant decrease in plasma insulin and associated rise in glucose concentration were observed. Fasted normal and leptin receptor-deficient db/db mice showed no response to leptin. A dose of leptin, which mimicked that found in normal mice, was administered to leptin-deficient, hyperinsulinemic ob/ob mice. This caused a marked lowering of plasma insulin concentration and a doubling of plasma glucose. Thus, leptin has a powerful acute inhibitory effect on insulin secretion. These results suggest that the action of leptin may be one mechanism by which excess adipose tissue could acutely impair carbohydrate metabolism.


Study of Amiloride Binding to Human Serum Albumin: Insights from Thermodynamic, Spectroscopic, and Molecular Docking Investigations.
This study was undertaken to investigate the interaction between the sodium channel blocker amiloride (AML) and human serum albumin (HSA). A combination of multi-spectroscopic techniques and computational methods were employed to identify the AML binding site on HSA and the forces responsible for the formation of the HSA-AML complex. Our findings revealed that AML specifically binds to Sudlow's site II, located in subdomain IIIA of HSA, and that the complex formed is stabilized using van der Waals hydrogen-bonding and hydrophobic interactions. FRET analysis showed that the distance between AML and Trp214 was optimal for efficient quenching. UV-Vis spectroscopy and circular dichroism indicated minor changes in the structure of HSA after AML binding, and molecular dynamics simulations (MDS) conducted over 100 ns provided additional evidence of stable HSA-AML-complex formation. This study enhances understanding of the interaction between AML and HSA and the mechanism responsible.


A comparison of gravid and under-house CO2-baited CDC light traps for mosquito species of public health importance in Houston, Texas.
The relative efficacy of gravid and under-house CO2 traps for monitoring mosquito species of public health importance within the Houston metroplex area was assessed. Gravid and under-house traps were colocated at 10 sites and monitored weekly between 1 March to 31 May 2007. The most numerous species caught was Culex pipiens quinquefasciatus Say. Other species of public health importance caught in gravid and under-house traps included Culex restuans Theobald, Aedes aegypti (L.), and Aedes albopictus Skuse. Adjusting for the week of collection, gravid traps caught significantly more mosquitoes (mean 23.1 per trap) in the study area than under-house traps (mean 3.6 per trap). However, under-house traps caught a greater variety of mosquito species (13) than gravid traps (11). Gravid and under-house traps only caught nine of 15 of the same mosquito species during the study period. In this study area, gravid traps should be used as the primary method of surveillance for mosquito-borne diseases of public health importance during the early part of the season, because of greater catch numbers of mosquitoes that pose a public health risk.


Insulin secretion during acid-base alterations.
Insulin secretion under extracellular acid-base alterations (metabolic acidosis or alkalosis) was studied, by challenging in vitro perfused sodium pentobarbital-anesthetized-rat pancreases with glucose, arginine, and tolbutamide. Under our experimental conditions, the amount of insulin released was lower at pH 7.8 than the amount corresponding to the pH 7.4 control, in spite of the agent used to stimulate the pancreas. The effect of pH 7.0 on insulin secretion, however, depends on the type and concentration of the stimulus used. It enhances the secretion elicited by glucose (6.6 mM) and glucose plus arginine (6.6 and 10 mM, respectively). On the other hand, it reduces the beta cell response to glucose plus tolbutamide (3.3 mM and 400 microgram/ml, respectively), whereas the response to high glucose (16.6 mM) is reduced in the first phase and not affected in the second. According to these results, modifications of the extracellular pH, mainly at high levels, may interfere with a common process involved in insulin secretion, namely beta cell emiocytosis.


The relationship of thyroid cancer with radiation exposure from nuclear weapon testing in the Marshall Islands.
The US nuclear weapons testing program in the Pacific conducted between 1946 and 1958 resulted in radiation exposure in the Marshall Islands. The potentially widespread radiation exposure from radio-iodines of fallout has raised concerns about the risk of thyroid cancer in the Marshallese population. The most serious exposures and its health hazards resulted from the hydrogen-thermonuclear bomb test, the Castle BRAVO, on March 1, 1954. Between 1993 and 1997, we screened 3,709 Marshallese for thyroid disease who were born before the BRAVO test. It was 60% of the entire population at risk and who were still alive at the time of our examinations. We diagnosed 30 thyroid cancers and found 27 other study participants who had been operated for thyroid cancer before our screening in this group. Fifty-seven Marshallese born before 1954 (1.5%) had thyroid cancer or had been operated for thyroid cancer. Nearly all (92%) of these cancers were papillary carcinoma. We derived estimates of individual thyroid dose proxy from the BRAVO test in 1954 on the basis of published age-specific doses estimated on Utirik atoll and 137Cs deposition levels on the atolls where the participants came from. There was suggestive evidence that the prevalence of thyroid cancer increased with category of estimated dose to the thyroid.


Microwave spectra, molecular structure, and aromatic character of 4a,8a-azaboranaphthalene.
The microwave spectra for seven unique isotopologues of 4a,8a-azaboranaphthalene [hereafter referred to as BN-naphthalene] were measured using a pulsed-beam Fourier transform microwave spectrometer. Spectra were obtained for the normal isotopologues with (10)B, (11)B, and all unique single (13)C and the (15)N isotopologue (with (11)B), in natural abundance. The rotational, centrifugal distortion and quadrupole coupling constants determined for the (11)B(14)N isotopologue are A = 3042.712 75(43) MHz, B = 1202.706 57(35) MHz, C = 862.220 13(35) MHz, DJ = 0.06(1) kHz, 1.5χaa ((14)N) = 2.5781(61) MHz, 0.25(χbb - χcc) ((14)N) = - 0.1185(17) MHz, 1.5χaa (11B) = - 3.9221(75) MHz, and 0.25(χbb - χcc) ((11)B) = - 0.9069(24) MHz. The experimental inertial defect is Δ = - 0.159 amu Å(2), which is consistent with a planar structure for the molecule. The B-N bond length from the experimentally determined structure is 1.47 Å, which indicates π-bonding character between the B and N. The measured quadrupole coupling strengths provide important and useful information about the bonding, orbital occupancy, and aromatic character for this aromatic molecule. Extended Townes-Dailey analyses were used to determine the B and N electron sp(2)-hybridized and p-orbital occupations. These results are compared with electron orbital occupations from the natural bond orbital option in theoretical calculations. From the analyses, it was determined that BN-naphthalene has aromatic character similar to that of other N-containing aromatics. The results are compared with similar results for B-N bonding in 1,2-dihydro-1,2-azaborine and BN-cyclohexene. Accurate and precise structural parameters were obtained from the microwave measurements on seven isotopologues and from high-level G09 calculations.


Long-term neuropathological and behavioral impairments after exposure to nerve agents.
One of the deleterious effects of acute nerve agent exposure is the induction of status epilepticus (SE). If SE is not controlled effectively, it causes extensive brain damage. Here, we review the neuropathology observed after nerve agent-induced SE, as well as the ensuing pathophysiological, neurological, and behavioral alterations, with an emphasis on their time course and longevity. Limbic structures are particularly vulnerable to damage by nerve agent exposure. The basolateral amygdala (BLA), which appears to be a key site for seizure initiation upon exposure, suffers severe neuronal loss; however, GABAergic BLA interneurons display a delayed death, perhaps providing a window of opportunity for rescuing intervention. The end result is a long-term reduction of GABAergic activity in the BLA, with a concomitant increase in spontaneous excitatory activity; such pathophysiological alterations are not observed in the CA1 hippocampal area, despite the extensive neuronal loss. Hyperexcitability in the BLA may be at least in part responsible for the development of recurrent seizures and increased anxiety, while hippocampal damage may underlie the long-term memory impairments. Effective control of SE after nerve agent exposure, such that brain damage is also minimized, is paramount for preventing lasting neurological and behavioral deficits.


Purification and characterisation of an extracellular serine proteinase from Aspergillus fumigatus and its detection in tissue.
A serine proteinase (Alp) from the culture supernate of a clinical isolate of Aspergillus fumigatus was purified to virtual homogeneity at a yield of 41%. The procedure involved affinity chromatography on agarose-epsilon-amino-caproyl-D-tryptophan methyl ester. Alp had an estimated mol. wt of 32 Kda and the pI was determined at pH 7.9. The enzyme was fully inhibited by phenylmethyl sulphonyl fluoride, chymostatin and alpha-1-proteinase inhibitor, and it was largely inhibited by alpha-1-anti-chymotrypsin. Partial inhibition was observed with tosyl-phenylalanine chloromethyl ketone, but tosyl-lysine chloromethyl ketone was ineffective. Thus, Alp may be identical with the major chymotryptic activity of A. fumigatus, which has already been described. The N-terminal sequence of 25 amino acids revealed an 88% homology of Alp with the subtilisin-related proteinase of A. oryzae. Alp acted on casein over a broad range from pH 5.5 to 11.5 and also acts to a lesser extent on haemoglobin and serum albumin. The enzyme degraded elastin and a synthetic elastase substrate; hence, it may be identical with the previously described elastinolytic activity of the fungus. At pH 7.3 and a concentration of 1 microgram/ml, Alp was not toxic for Vero cells, but it efficiently detached such cells from a plastic surface. Specific antibodies against Alp were detected by enzyme immunoassay in the sera of patients and Alp-antigen was demonstrated by immunofluorescence in mycotic human lung. In addition, a second proteinase (Exalp) with extremely alkaline activity, and an aspartic proteinase of A. fumigatus are described.


Neonatal kaempferol exposure attenuates impact of cerebral palsy model on neuromotor development, cell proliferation, microglia activation, and antioxidant enzyme expression in the hippocampus of rats.
This study aims to assess the effect of neonatal treatment with kaempferol on neuromotor development, proliferation of neural precursor cells, the microglia profile, and antioxidant enzyme gene expression in the hippocampus. A rat model of cerebral palsy was established using perinatal anoxia and sensorimotor restriction of hindlimbs during infancy. Kaempferol (1 mg/ kg) was intraperitoneally administered during the neonatal period. Neonatal treatment with kaempferol reduces the impact of the cerebral palsy model on reflex ontogeny and on the maturation of physical features. Impairment of locomotor activity development and motor coordination was found to be attenuated by kaempferol treatment during the neonatal period in rats exposed to cerebral palsy. Neonatal treatment of kaempferol in cerebral palsy rats prevents a substantial reduction in the number of neural precursor cells in the dentate gyrus of the hippocampus, an activated microglia profile, and increased proliferation of microglia in the sub-granular zone and in the granular cell layer. Neonatal treatment with kaempferol increases gene expression of superoxide dismutase and catalase in the hippocampus of rats submitted to the cerebral palsy model. Kaempferol attenuates the impact of cerebral palsy on neuromotor behavior development, preventing altered hippocampal microglia activation and mitigating impaired cell proliferation in a neurogenic niche in these rats. Neonatal treatment with kaempferol also increases antioxidant defense gene expression in the hippocampus of rats submitted to the cerebral palsy model.


The central morphology of mechanoreceptor afferents in the metathoracic leg of the cockroach, Periplaneta americana (Insecta).
The sensory arborizations of the femoral chordotonal organ (FECO), trochanteral campaniform sensilla (CS) and the trochanteral hairplate (THP) are restricted to the ipsilateral hemiganglion and possess large dendritic fields in the dorsal and ventral neuropiles. The efferent projections of the fast (Df) and the slow (Ds) depressor motoneurons project into the same areas. Three groups of trichoid sensilla located on the coxal and trochanteral cuticular walls have central contralateral projections and in one case intersegmental projections. All sensory projections terminate either just lateral of or in the Ventral Association Centre (VAC). Comparisons with the mechanoreceptors of other insects reveal similar projection patterns for some proprioceptors and non-homologous central configurations for others.


Barriers and facilitators of risk-based health care for adult survivors of childhood cancer: A report from the Childhood Cancer Survivor Study.
Optimal risk-based survivor health care includes surveillance for late effects and education targeted at reducing or preventing risky health behaviors. Understanding the reasons for a lack of risk-based follow-up care is essential. Adult participants from the Childhood Cancer Survivor Study were surveyed about having a cancer-related visit in the past 2 years and the likelihood of having a cancer-related visit in the future. Additional factors thought to be related to the primary outcomes were also assessed. Nine hundred seventy-five survivors completed the survey. Twenty-seven percent (95% confidence interval [CI], 24%-30%) had a cancer-related medical visit in the previous 2 years, and 41% (95% CI, 38%-44%) planned to have such a visit within the next 2 years. The likelihood of having had a cancer-related visit within the last 2 years was higher among survivors assigning greater importance to these visits (relative risk [RR], 1.2; 95% CI, 1.1-1.3), perceiving greater susceptibility to health problems (RR, 1.2; 95% CI, 1.1-1.3), having a moderate to life-threatening chronic health problem related to their cancer (RR, 2.1; 95% CI, 1.7-2.7), seeing a primary care provider for a cancer-related problem (RR, 1.3; 95% CI, 1.0-1.6), having a cancer treatment summary (RR, 1.3; 95% CI, 1.0-1.6), and endorsing greater confidence in physicians' abilities to address questions and concerns (RR, 1.2; 95% CI, 1.0-1.3). Educational interventions improving awareness of treatment history and susceptibility to cancer-related late effects and corresponding risk-based care are likely to be beneficial for survivors of childhood cancers.


Characterization of Bacteria and Inducible Phages in an Intensive Care Unit.
Intensive care units (ICUs) are critical locations for the transmission of pathogenic and opportunistic microorganisms. Bacteria may develop a synergistic relationship with bacteriophages and more effectively resist various stresses, enabling them to persist despite disinfection and antimicrobial treatment. We collected 77 environmental samples from the surroundings of 12 patients with infection/colonizations by Escherichia coli, Staphylococcus aureus or Klebsiella spp in an ICU in Austria. Surface swabs were tested for lytic phages and bacterial isolates for mitomycin C-inducible prophages. No lytic bacteriophages were detected, but S. aureus was isolated from the surroundings of all patients. About 85% of the colonies isolated from surface samples were resistant to antimicrobials, with 94% of them multidrug resistant. Two inducible temperate bacteriophages-myovirus vB_EcoM_P5 and siphovirus vB_SauS_P9-were recovered from two clinical isolates. Staphylococci phage vB_SauS_P9 lysed S. aureus isolates from the surface swabs collected from the surroundings of three patients. No transductants were obtained on propagation in phage-sensitive antimicrobial-resistant isolates. The two phages were sensitive to 0.25% (v/v) of the disinfectant TPH Protect, which eliminated viable phages after 15 min. Coliphage vB_EcoM_P5 was inactivated at 70 °C and staphylococci phage vB_SauS_P9 at 60 °C after 60 min.


Reporting of conflicts of interest in meta-analyses of trials of pharmacological treatments.
Disclosure of conflicts of interest (COIs) from pharmaceutical industry study funding and author-industry financial relationships is sometimes recommended for randomized controlled trials (RCTs) published in biomedical journals. Authors of meta-analyses, however, are not required to report COIs disclosed in original reports of included RCTs. To investigate whether meta-analyses of pharmacological treatments published in high-impact biomedical journals report COIs disclosed in included RCTs. We selected the 3 most recent meta-analyses of patented pharmacological treatments published January 2009 through October 2009 in each general medicine journal with an impact factor of at least 10; in high-impact journals in each of the 5 specialty medicine areas with the greatest 2008 global therapeutic sales (oncology, cardiology, respiratory medicine, endocrinology, and gastroenterology); and in the Cochrane Database of Systematic Reviews. Two investigators independently extracted data on disclosed study funding, author-industry financial ties, and author employment from each meta-analysis, from RCTs included in each meta-analysis, and on whether meta-analyses reported disclosed COIs of included RCTs. Of 29 meta-analyses reviewed, which included 509 RCTs, only 2 meta-analyses (7%) reported RCT funding sources; and 0 reported RCT author-industry ties or employment by the pharmaceutical industry. Of 318 meta-analyzed RCTs that reported funding sources, 219 (69%) were industry funded; and 91 of 132 (69%) that reported author financial disclosures had 1 or more authors with pharmaceutical industry financial ties. In 7 of the 29 meta-analyses reviewed, 100% of included RCTs had at least 1 form of disclosed COI (pharmaceutical industry funding, author-industry financial ties, or employment), yet only 1 of these 7 meta-analyses reported RCT funding sources, and 0 reported RCT author-industry ties or employment. Among a group of meta-analyses of pharmacological treatments published in high-impact biomedical journals, information concerning primary study funding and author COIs for the included RCTs were only rarely reported.


The lived experience of fathers whose children are diagnosed with a genetic disorder.
To describe the father's experience of having a child diagnosed with a genetic disorder. Interpretive phenomenological analysis. In-person interviews for participants living within 100 miles of the researcher; phone interviews with participants living more than 100 miles from the researcher. Six fathers of children diagnosed with genetic disorders. Participants were recruited with the assistance of nurses, physicians, and genetic counselors. Snowball sampling, Internet and e-mail recruitment, and word of mouth were also used. Semistructured interviews were completed, audio recorded, and transcribed verbatim. Five themes emerged: the impact of diagnosis, seeking understanding, coping with effects of the disorder, looking to the future, and addressing a father's needs. Fathers of children diagnosed with genetic disorders described a lack of education and information regarding the diagnoses. They attributed this deficiency to a lack of knowledge among physicians, nurses, and other health care professionals. Coping with the effects of the disorder was identified as an integral part of adapting to meet the child's needs. Understanding the experience of a father whose child is diagnosed with a genetic disorder is necessary to ensure provision of proper intervention and care.


Musical hallucinations and triazolam use.
Musical hallucinations developed in a healthy 57-year-old man after using hypnotic triazolam (Halcion) for eight nights; the hallucinations continued for over one year. Although a causal relationship to triazolam cannot be proved, use of this drug has been associated with other bizarre alterations of memory.


Three-dimensional tissue model in direct contact with an on-chip vascular bed enabled by removable membranes.
Three-dimensional (3D) tissue culture is a powerful tool for understanding physiological events. However, 3D tissues still have limitations in their size, culture period, and maturity, which are caused by the lack of nutrients and oxygen supply through the vasculature. Here, we propose a new method for culturing a 3D tissue-a spheroid-directly on an 'on-chip vascular bed'. The method can be applied to any 3D tissue because the vascular bed is preformed, so that angiogenic factors from the tissue are not necessary to induce vasculature. The essential component of the assay system is the removable membrane that initially separates the 3D tissue culture well and the microchannel in which a uniform vascular bed is formed, and then allows the tissue to be settled directly onto the vascular bed following its removal. This in vitro system offers a new technique for evaluating the effects of vasculature on 3D tissues.


The capability of heterogeneous γδ T cells in cancer treatment.
γδ T cells, a specialized subset of T lymphocytes, have garnered significant attention within the realm of cancer immunotherapy. Operating at the nexus between adaptive and innate immunological paradigms, these cells showcase a profound tumor discernment repertoire, hinting at novel immunotherapeutic strategies. Significantly, these cells possess the capability to directly identify and eliminate tumor cells without reliance on HLA-antigen presentation. Furthermore, γδ T cells have the faculty to present tumor antigens to αβ T cells, amplifying their anti-tumoral efficacy.Within the diverse and heterogeneous subpopulations of γδ T cells, distinct immune functionalities emerge, manifesting either anti-tumor or pro-tumor roles within the tumor microenvironment. Grasping and strategically harnessing these heterogeneous γδ T cell cohorts is pivotal to their integration in tumor-specific immunotherapeutic modalities. The aim of this review is to describe the heterogeneity of the γδ T cell lineage and the functional plasticity it generates in the treatment of malignant tumors. This review endeavors to elucidate the intricate heterogeneity inherent to the γδ T cell lineage, the consequential functional dynamics in combating malignancies, the latest advancements from clinical trials, and the evolving landscape of γδ T cell-based oncological interventions, while addressing the challenges impeding the field.


Signalling mechanisms in progesterone-neurotransmitter interactions.
The current and expanded view of transcriptional regulation by the steroid/thyroid superfamily of nuclear transcription factors integrates not only the hormone-dependent but also the hormone-independent cellular signaling mechanisms in physiology and reproduction. This effort has vastly been aided by the identification of steroid hormone receptors as transcriptional mediators of a variety of ligands, whose transcriptional response is dependent upon cross-talk with distinct signal transduction pathways, their recruitment of coregulators, alteration of chromatin structure and identification of specific interactive motifs within the receptors themselves. This review will provide a framework for the current concepts in the field of steroid hormone action as exemplified by our studies on progesterone receptor in female sexual behavior.


Incorporating mutational heterogeneity to identify genes that are enriched for synonymous mutations in cancer.
Synonymous mutations, which change the DNA sequence but not the encoded protein sequence, can affect protein structure and function, mRNA maturation, and mRNA half-lives. The possibility that synonymous mutations might be enriched in cancer has been explored in several recent studies. However, none of these studies control for all three types of mutational heterogeneity (patient, histology, and gene) that are known to affect the accurate identification of non-synonymous cancer-associated genes. Our goal is to adopt the current standard for non-synonymous mutations in an investigation of synonymous mutations. Here, we create an algorithm, MutSigCVsyn, an adaptation of MutSigCV, to identify cancer-associated genes that are enriched for synonymous mutations based on a non-coding background model that takes into account the mutational heterogeneity across these levels. Using MutSigCVsyn, we first analyzed 2572 cancer whole-genome samples from the Pan-cancer Analysis of Whole Genomes (PCAWG) to identify non-synonymous cancer drivers as a quality control. Indicative of the algorithm accuracy we find that 58.6% of these candidate genes were also found in Cancer Census Gene (CGC) list, and 66.2% were found within the PCAWG cancer driver list. We then applied it to identify 30 putative cancer-associated genes that are enriched for synonymous mutations within the same samples. One of the promising gene candidates is the B cell lymphoma 2 (BCL-2) gene. BCL-2 regulates apoptosis by antagonizing the action of proapoptotic BCL-2 family member proteins. The synonymous mutations in BCL2 are enriched in its anti-apoptotic domain and likely play a role in cancer cell proliferation. Our study introduces MutSigCVsyn, an algorithm that accounts for mutational heterogeneity at patient, histology, and gene levels, to identify cancer-associated genes that are enriched for synonymous mutations using whole genome sequencing data. We identified 30 putative candidate genes that will benefit from future experimental studies on the role of synonymous mutations in cancer biology.


An environmental scan of residential treatment service provision in Ontario.
Ontario has one of the highest rates of substance-related harms in Canada. Residential treatment programs in the province provide a variety of in-house treatment services to support the needs of individuals with substance use disorders (SUD). However, these programs are not standardized, often varying in the type, quality, and availability of services offered, including evidence-based interventions such as Opioid Agonist Treatment (OAT). Local treatment systems are also rather fragmented and complex to navigate, creating barriers for potential services users to identify and make informed choices on available treatment options. Between May to August 2023, we conducted an environmental scan to capture available information on all publicly-funded residential treatment programs in Ontario using the ConnexOntario service portal, a government-funded, health services information platform. Data were captured on organization name, geographical location, program description, program type (residential addictions treatment or supportive recovery programs), eligibility criteria, target population, the program's OAT policies, number of available beds, minimum and maximum length of stay, projected wait times, funding source, and associated fees for program admission. Data were extracted and organized by geographic region, and findings were presented descriptively. A total of 102 residential addiction treatment programs and 36 residential supportive recovery programs in Ontario were identified. The scan noted substantial regional variations in program availability and wait times, along with a lack of programs tailored to unique populations such as women, youth, and Indigenous peoples. There is also a paucity of publicly-available information on program offerings, including detailed specifics on OAT policies within residential treatment programs that are crucial to ensuring that the services being offered are safe and grounded in evidence-based practice. Findings from the scan highlight notable gaps in program types, offerings, and availability among residential treatment programs in the province, including a lack of standardization on OAT policies across programs. Efforts should be made to ensure access to treatment-specific program information relevant to potential service users and to enhance coordinated access to residential treatment services in the province.


Transcriptional Response to Acute Thermal Exposure in Juvenile Chinook Salmon Determined by RNAseq.
Thermal exposure is a serious and growing challenge facing fish species worldwide. Chinook salmon (Oncorhynchus tshawytscha) living in the southern portion of their native range are particularly likely to encounter warmer water due to a confluence of factors. River alterations have increased the likelihood that juveniles will be exposed to warm water temperatures during their freshwater life stage, which can negatively impact survival, growth, and development and pose a threat to dwindling salmon populations. To better understand how acute thermal exposure affects the biology of salmon, we performed a transcriptional analysis of gill tissue from Chinook salmon juveniles reared at 12° and exposed acutely to water temperatures ranging from ideal to potentially lethal (12° to 25°). Reverse-transcribed RNA libraries were sequenced on the Illumina HiSeq2000 platform and a de novo reference transcriptome was created. Differentially expressed transcripts were annotated using Blast2GO and relevant gene clusters were identified. In addition to a high degree of downregulation of a wide range of genes, we found upregulation of genes involved in protein folding/rescue, protein degradation, cell death, oxidative stress, metabolism, inflammation/immunity, transcription/translation, ion transport, cell cycle/growth, cell signaling, cellular trafficking, and structure/cytoskeleton. These results demonstrate the complex multi-modal cellular response to thermal stress in juvenile salmon.


Functional characterization of 23 allelic variants of thiopurine S-methyltransferase gene (TPMT*2 - *24).
Thiopurine S-methyltransferase (TPMT) is an enzyme responsible for the detoxification of the widely used thiopurine drugs. TPMT is genetically polymorphic and is associated with large interindividual variations in thiopurine drug toxicity and therapeutic efficacy. In this study, we performed an in-vitro analysis of TPMT variant alleles, namely, TPMT*2, *3A, *3B, *3C, *5, *6, *7, *8, *9, *10, *11, *12, *13, *14, *16, *17, *18, *19, *20, *21, *22, *23, and *24. The wild-type TPMT proteins, TPMT.1 and 23 variants were heterologously expressed in COS-7 cells, and the kinetic parameters Km, Vmax, and intrinsic clearance (Vmax/Km) of 6-thioguanine S-methylation were determined. The expression levels of TPMT.2, TPMT.3A, TPMT.5, TPMT.12, TPMT.14, and TPMT.22 were considerably lower than that of TPMT.1 (P<0.005), and that of TPMT.18 was slightly reduced (P<0.05). The kinetic parameters of TPMT.3A, TPMT.3B, TPMT.5, TPMT.14, TPMT.18, TPMT.21, and TPMT.22 could not be accurately established because of no activity in 6-thioguanine S-methylation. The Vmax/Km values of TPMT.2, TPMT.7, TPMT.17, and TPMT.24 were displayed less than 10% of the wild-type. This functional analysis with respect to TPMT variants could provide useful information for individualization of thiopurine drugs therapy.


Antimicrobial use and the influence of inadequate empiric antimicrobial therapy on the outcomes of nosocomial bloodstream infections in a neonatal intensive care unit.
To evaluate antimicrobial use and the influence of inadequate empiric antimicrobial therapy on the outcomes of nosocomial bloodstream infections (BSIs). Prospective cohort study with nested case-control analysis. Neonatal intensive care unit (NICU). All patients weighing 2,000 g or less were enrolled. Data collection included risk factors for nosocomial BSI, admission severity of illness, microbiology, antimicrobial therapy, and outcomes. Inadequate empiric antimicrobial therapy was defined as the use of antibiotics for more than 48 hours after the day that blood cultures were performed that did not cover the microorganisms causing the bacteremia or administration of antibiotics that failed to cover resistant microorganisms. Two hundred twenty-nine patients were enrolled. Forty-five developed nosocomial BSIs. The BSI rates were 11.2, 2.8, and 0 per 1,000 catheter-days for patients weighing 1,000 g or less, between 1,001 and 1,500 g, and between 1,501 and 2,000 g, respectively. After adjustment for severity of illness, the mortality in patients with nosocomial BSI receiving inadequate empiric antimicrobial therapy was higher than in those receiving adequate therapy (adjusted odds ratio [AOR], 5.3; 95% confidence interval [CI95], 1.2-23.2). By multivariate analysis, nosocomial BSI attributed to Candida species (AOR, 6.3; CI95, 1.4-28.0) and invasive procedure prior to onset of BSI (AOR, 6.4; CI95, 1.0-39.0) were associated with administration of inadequate empiric antimicrobial therapy. Administration of inadequate empiric antimicrobial therapy among NICU patients with nosocomial BSI was associated with higher mortality. Additional studies on the role of inadequate empiric antimicrobial therapy and the outcomes of BSIs among NICU patients are needed.


Statistical methods for body mass index: A selective review.
Obesity rates have been increasing over recent decades, causing significant concern among policy makers. Excess body fat, commonly measured by body mass index, is a major risk factor for several common disorders including diabetes and cardiovascular disease, placing a substantial burden on health care systems. To guide effective public health action, we need to understand the complex system of intercorrelated influences on body mass index. This paper, based on all eligible articles searched from Global health, Medline and Web of Science databases, reviews both classical and modern statistical methods for body mass index analysis. We give a description of each of these methods, exploring the classification, links and differences between them and the reasons for choosing one over the others in different settings. We aim to provide a key resource and statistical library for researchers in public health and medicine to deal with obesity and body mass index data analysis.


Serum cystatin-C and BETA 2-microglobulin as accurate markers in the early diagnosis of kidney injury in neonates: A single center study.
Early detection of kidney injury in neonates is very important for appropriate management and prevention of serious complications; however, commonly used detectors as creatinine and blood urea nitrogen (BUN) do not directly reflect early renal cell injury. Serum cystatin-C (Cys-C) and beta 2-microglobulin (Β2M), serum creatinine and BUN were assessed in 20 neonates who developed renal impairment after admission to the neonatal intensive care unit and 10 healthy neonates. The means of serum Cys-C, Β2M and creatinine on Day 1 of admission in the cases and control groups were 2.15 ± 0.52 vs 0.45 ± 0.19, 7.18 ± 2.36 vs 1.92 ± 0.41 and 0.81 ± 0.07 vs 0.53 ± 0.20, respectively, with P-value <0.05 in only Cys-C and Β2M. We conclude that serum Cys-C and Β2M are suggested as simple and accurate markers for the early diagnosis of kidney injury in neonates than serum creatinine.


Identification of a gene essential for the first committed step in the biosynthesis of bacteriochlorophyll c.
Bacteriochlorophylls (BChls) c, d, and e are the major chlorophylls in chlorosomes, which are the largest and one of the most efficient antennae produced by chlorophototrophic organisms. In the biosynthesis of these three BChls, a C-13(2)-methylcarboxyl group found in all other chlorophylls (Chls) must be removed. This reaction is postulated to be the first committed step in the synthesis of these BChls. Analyses of gene neighborhoods of (B)Chl biosynthesis genes and distribution patterns in organisms producing chlorosomes helped to identify a gene (bciC) that appeared to be a good candidate to produce the enzyme involved in this biochemical reaction. To confirm that this was the case, a deletion mutant of an open reading frame orthologous to bciC, CT1077, was constructed in Chlorobaculum tepidum, a genetically tractible green sulfur bacterium. The CT1077 deletion mutant was unable to synthesize BChl c but still synthesized BChl a and Chl a. The deletion mutant accumulated large amounts of various (bacterio)pheophorbides, all of which still had C-13(2)-methylcarboxyl groups. A C. tepidum strain in which CT1077 was replaced by an orthologous gene, Cabther_B0081 [corrected] from "Candidatus Chloracidobacterium thermophilum" was constructed. Although the product of Cabther_B0081 [corrected] was only 28% identical to the product of CT1077, this strain synthesized BChl c, BChl a, and Chl a in amounts similar to wild-type C. tepidum cells. To indicate their roles in the first committed step of BChl c, d, and e biosynthesis, open reading frames CT1077 and Cabther_B0081 [corrected] have been redesignated bciC. The potential mechanism by which BciC removes the C-13(2)-methylcarboxyl moiety of chlorophyllide a is discussed.


Moderate Depression Promotes Posttraumatic Growth (Ptg): A Young Population Survey 2 Years after the 2009 L'Aquila Earthquake.
Earthquakes can result in a range of psychopathology and in negative and positive consequences for survivors. To examine the association between clinical aftereffects (anxiety and depressive symptoms) and post-traumatic growth (PTG) among young survivors of the 2009 L'Aquila earthquake, Italy. 316 young earthquake survivors enrolled in the University of L'Aquila were evaluated two years after the natural disaster. Participants completed three main questionnaires, including Patient Health Questionnaire-9 items (PHQ-9), Self-Rating Anxiety Scale (SAS), and Posttraumatic Growth Inventory (PTGI). 59.6% of the student sample showed different levels of depression, whereas 13.3% reported anxiety symptoms. In both clinical dimensions (anxiety and depression), gender differences were found: female gender was confirmed risk factor for a clinical post-traumatic response. Personal PTG, demonstrated by 18% of the L'Aquila youths included in our sample, was predicted by moderate levels of depression (O.R. 2.7). In our model, gender, age, and anxiety did not show any predictive value. In a post-traumatic setting, the development of individual cognitive strategies is crucial, whereas after a natural disaster, paradoxically, a moderate depressive condition and the related distress could promote the drive to overcome the psychological consequences of the traumatic event.


Cardiovascular factors influencing survival in dialysis patients.
Cardiovascular disease is the most common cause of death among dialysis patients. Cardiovascular abnormalities that predispose to mortality include acute myocardial infarction, chronic ischemic heart disease, atherosclerotic peripheral vascular disease, congestive heart failure, left ventricular hypertrophy, dilated cardiomyopathy, hypertrophic hyperkinetic heart disease, prior cardiac arrest, and possibly systemic hypertension. This review explores the influences of cardiovascular disease on survival in patients with end-stage renal disease treated with dialysis.


Alteration of luminal membrane structure by antidiuretic hormone.
The final step in the action of antidiuretic hormone (ADH) is the insertion of aggregates of water-conducting particles into the luminal membrane of the receptor cell. The aggregates arise from cytoplasmic tubular structures that fuse with the luminal membrane. This review presents a number of questions about this process, along with current attempts to answer them. The following topics are addressed: 1) the exact role of the cytoskeleton in promoting tubular fusion, 2) the nature of the translocation process leading to fusion, 3) the point in the sequence at which ADH enters, 4) the composition and structure of the water-conducting particles. Some of the answers require a more complete understanding of the structure of the apical portion of ADH-sensitive cells; recent morphological studies are therefore included.


Orbital muscle of Müller: observations on human fetuses measuring 35-150 mm.
The arrangement and relationships of the orbital muscle of Müller in human fetuses have been analyzed. This is a constant muscle made of smooth muscle fibers arranged over the longer axis of the inferior orbital fissure; some of its fibers reach the inferior wall of the cavernous sinus. The muscle layer is pierced by orbital rami of the medial maxillary artery and by thin veins communicating with the ophthalmic vein system and pterygoid plexus. The zygomatic nerve can be found in the midst of the muscle mass along most of its course. It is innervated by short rami from the sphenopalatine ganglion.


The protein disulphide-isomerase family: unravelling a string of folds.
The mammalian protein disulphide-isomerase (PDI) family encompasses several highly divergent proteins that are involved in the processing and maturation of secretory proteins in the endoplasmic reticulum. These proteins are characterized by the presence of one or more domains of roughly 95-110 amino acids related to the cytoplasmic protein thioredoxin. All but the PDI-D subfamily are composed entirely of repeats of such domains, with at least one domain containing and one domain lacking a redox-active -Cys-Xaa-Xaa-Cys- tetrapeptide. In addition to their known roles as redox catalysts and isomerases, the last few years have revealed additional functions of the PDI proteins, including peptide binding, cell adhesion and perhaps chaperone activities. Attention is now turning to the non-redox-active domains of the PDIs, which may play an important role in all of the known activities of these proteins. Thus the presence of both redox-active and -inactive domains within these proteins portends a complexity of functions differentially accommodated by the various family members.


[Evaluation of the state of the progeny of mice with damaged kidneys].
It was established that increased mortality was characteristic of newborn mice from females with dystrophic lesions of the renal tissue. The kidneys of newborns from these females had a less relative weight as compared with newborns from healthy mice. Other organs of experimental and control newborns did not differ in their relative weight. Hypertrophy of the tubule cells with the signs of picnosis of nuclei was noted in the kidneys of experimental newborn animals. In two-month-old mice of the experimental and control groups the kidneys and other organs (liver, heart, lungs, spleen) had no substantial distinctions in the relative weight. The concentration of urea in the blood of two-month-old mice from females with injured kidneys under protein load was higher than in control two-month-old mice from females treated with 0,85% solution of NaCl which speaks of decreased resistance of kidneys in the animals of experimental group against pathogenic factors.


Involvement of Activation of Mitogen-Activated Protein Kinase (MAPK)/Extracellular Signal-Regulated Kinase (ERK) Signaling Pathway in Proliferation of Urethral Plate Fibroblasts in Finasteride-Induced Rat Hypospadias.
BACKGROUND We investigated the role of the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) signaling pathway in finasteride-induced hypospadias rats and explored the mechanisms involved. MATERIAL AND METHODS The hypospadias model was established by intragastric administration of finasteride and confirmed by hematoxylin and eosin (HE) staining. The urethral plate fibroblasts (UPF) were obtained from normal and modeled rats and identified based upon vimentin expression. Thereafter, UPF were divided into a normal control group, a model group, a model + MAPK inhibitor group, and a model + ERK inhibitor group. Cell proliferation, apoptosis, and cell cycling of UPF were assessed. Quantitative real-time PCR and Western blot analysis were used to evaluate expression of the MAPK signaling pathway and apoptosis-related genes. RESULTS HE staining confirmed that 10 mg/kg finasteride caused severe hypospadias in rats. UPFs obtained from the 10 mg/kg finasteride group showed higher proliferation and cell cycling and lower apoptosis compared with those obtained from the normal control group (P<0.05). Interestingly, a MAPK inhibitor or an ERK inhibitor could attenuate the abnormalities of cell proliferation, cycling, and apoptosis of UPF induced by finasteride. Compared with controls, the relative expression of p-MEK1/MEK1, caspase 3, and P53 in the UPF of the model group were reduced, while the relative expression of p-MAPK14/MAPK14 was increased in the cells of the model group. By contrast, a MAPK inhibitor or an ERK inhibitor could alleviate the abnormalities of MAPK/ERK signaling pathway and apoptosis-related gene expression induced by finasteride. CONCLUSIONS Our study reveals that the MAPK/ERK signaling pathway is involved in the regulation of proliferation, apoptosis, and cell cycling of UPFs in finasteride-induced hypospadias.


Development and Testing of the CAHPS Cancer Care Survey.
The Consumer Assessment of Healthcare Providers and Systems (CAHPS) Cancer Care Survey is a systematic assessment of health care experiences of patients with cancer. It supports comparisons among all cancer treatment settings and modalities. Formative research included 16 focus groups with patients receiving treatment and family members; advice from a panel of oncology and quality improvement experts; and interviews with stakeholders representing oncology associations, accredited cancer centers, and community oncology practices. We conducted cognitive tests of the instrument and field tests at six cancer centers and four community oncology practices, after which the survey was finalized and obtained the CAHPS trademark. The survey includes 56 questions that form six core composite measures (Getting Timely Care; Supporting Patient Self-Management; Available to Provide Care and Information; Provider Communication; Care Coordination; and Courteous Office Staff); two single-item measures of family participation in care and interpreter services; and two global ratings of cancer care and the treatment team. Sixteen additional items form three supplemental composite measures: Shared Decision-Making, Keeping Patients Informed, and Access to Care. Mail-only, mail-telephone mixed-mode, and Web-mail mixed-mode data collection methods are recommended. The questionnaires and instructions for use are free and available in English and Spanish on the CAHPS Website (www.ahrq.gov/cahps).


Flow diverter stents for unruptured saccular anterior circulation perforating artery aneurysms: safety, efficacy, and short-term follow-up.
Anterior circulation perforating artery aneurysms including anterior choroidal artery and lenticulostriate artery aneurysms are rare. Injury to these vessels can lead to severe debilitating symptoms. To present a new approach to treatment using flow diversion technology. Patients treated with a Pipeline embolization device (PED) for perforator artery aneurysms at our institution between June 2012 and May 2013 were identified and included in our retrospective analysis. We evaluated patient vascular risk factors; family history of aneurysms; aneurysm characteristics; National Institutes of Health Stroke Scale (NIHSS) and modified Rankin Scale (mRS) on admission; and angiography follow-up and patient clinical outcome at discharge, 6 months, and 1 year. We included four patients with a mean age of 59.8 years. Two patients had a positive family history of aneurysms. Patient vascular risk factors included smoking, dyslipidemia, and hypertension. All patients presented with a NIHSS and mRS of 0 on admission. Aneurysms were located at the anterior choroidal (n=2) or lenticulostriate artery (n=2) and were treated with a single PED. No periprocedural or postprocedural complications occurred. The patients were discharged with no change in NHISS or mRS score. Six-month and 1-year follow-up angiography showed complete aneurysm occlusion. Mild intimal hyperplasia was seen in 2 cases at 6 months, but was resolved at the 1-year follow-up. No re-treatment was necessary. NIHSS and mRS remained 0 at follow-up time points. Our preliminary results show that flow diversion technology is an effective and safe therapy for complex, hard-to-treat aneurysms in perforating arteries. Larger studies with long-term follow-up are needed to validate our promising results.


The Yin Yang 1 of Penaeus vannamei regulates transcription of the small subunit hemocyanin gene during Vibrio parahaemolyticus infection.
Hemocyanin is a respiratory protein, it is also a multifunctional immune molecule that plays a vital role against pathogen invasion in shrimp. However, the regulation of hemocyanin gene expression in shrimp hemocytes and the mechanisms involved during pathogen infection remains unclear. Here, we used DNA pull-down followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify the Yin Yang 1 transcription factor homolog in Penaeus vannamei (PvYY1) as a key factor that modulates transcription of the small subunit hemocyanin gene of P. vannamei (PvHMCs) in hemocytes during Vibrio parahaemolyticus AHPND (VPAHPND) infection. Bioinformatics analysis revealed that the core promoter region of PvHMCs contains two YY1 motifs. Mutational and oligoprecipitation analyses confirmed that PvYY1 could bind to the YY1 motifs in the PvHMCs core promoter region, while truncation of PvYY1 revealed that the N-terminal domain of PvYY1 is essential for the transactivation of PvHMCs core promoter. Besides, the REPO domain of PvYY1 could repress the activity of the PvHMCs core promoter. Overexpression of PvYY1 significantly activates the promoter activity of PvHMCs core promoter, while PvYY1 knockdown significantly decreases the expression level of PvHMCs in shrimp hemocytes and survival rate of shrimp upon infection with VPAHPND. Our present study provides new insights into the transcriptional regulation of PvHMCs by PvYY1 in shrimp hemocytes during bacteria (VPAHPND) infection.


How do GPs and patients share the responsibility for cancer safety netting follow-up actions? A qualitative interview study of GPs and patients in Oxfordshire, UK.
To explore patients' and General Practitioners' (GPs) accounts of how responsibility for follow-up was perceived and shared in their experiences of cancer safety netting occurring within the past 6 months. In-depth interviews were recorded and transcribed verbatim. Data were analysed through an abductive process, exploring anticipated and emergent themes. Conceptualisations of 'responsibility' were explored by drawing on a transactional to interdependent continuum drawing from the shared decision-making literature. A purposive sample of 25 qualified GPs and 23 adult patients in Oxfordshire, UK. The transactional sharing approach involves responsibility being passed from GP to patient. Patients expected and were willing to accept responsibility in this way as long as they received clear guidance from their GP and had capacity. In interdependent sharing, GPs principally aimed to reach consensus and share responsibility with the patient by explaining their rationale, uncertainty or by stressing the potential seriousness of the situation. Patients sharing this responsibility could be put at risk if no follow-up or timeframe was suggested, they had inadequate information, were falsely reassured or their concerns were not addressed at re-consultation. GPs and patients exchange and share responsibility using a combination of transactional and interdependent styles, tailoring information based on patient characteristics and each party's level of concern. Clear action plans (written where necessary) at the end of every consultation would help patients decide when to re-consult. Further research should investigate how responsibility is shared within and outside the consultation, within primary care teams and with specialist services.


The Fertility of Frozen Boar Sperm When used for Artificial Insemination.
One of the limits to practical use of frozen boar sperm involves the lowered fertility when used for artificial insemination. Years of studies have shown that 5-6 billion sperm (approximately 3 billion viable) used in single or multiple inseminations results in pregnancy rates most often between 60 and 70% and with litter sizes between nine and 10 pigs. Yet today, it is not uncommon for studies to report pregnancy rates from 70 to 85% and litter sizes with 11-12 pigs. While global statements about the incidence and reasons for higher fertility are not conclusive, incremental fertility improvements appear independently associated with use of a minimum number of viable sperm (1-2 billion), insemination timing that increases the probability that sperm will be present close to ovulation for groups of females, selection for boar sperm survival following cryopreservation, and modification of the freeze and thaw conditions using additives to protect sperm from oxidative damage. Studies show that techniques such as intrauterine and deep uterine insemination can provide an opportunity to reduce sperm numbers and that control of time of ovulation in groups of females can reduce the need for multiple inseminations and improve the chance for AI close to ovulation. However, optimal and consistent fertility with cryopreserved boar sperm may require a multifaceted approach that includes boar selection and screening, strategic use of additives during the freezing and thawing process, post-thaw evaluation of sperm and adjustments in sperm numbers for AI, assessment of female fertility and ovulation induction for single insemination. These sequenced procedures should be developed and incorporated into a quality control system for improved fertility when using minimal numbers of cryopreserved boar sperm.


Early time-course of respiratory mechanics, mechanical power and gas exchange in ARDS patients.
To describe the clinical course of ARDS during the first three days of mechanical ventilation, to compare ventilatory setting, respiratory mechanics and gas exchange variables collected during the first three days of mechanical ventilation between patients who survived and died during intensive care unit (ICU) stay and to investigate the variables associated with mortality at ICU admission and throughout the first three days of mechanical ventilation. Prospective observational study. Mechanically ventilated ARDS patients were studied at ICU admission and for the following three days. Univariate logistic regression models were performed for PaO2/FiO2 ratio, driving pressure and alveolar dead space fraction and for mechanical power and mechanical power ratio. Mechanical power ratio was higher in non survivors at ICU admission and over time; PaO2/FiO2 ratio was higher in survivors with a similar behavior over time in the two groups while alveolar dead space fraction was similar at ICU admission and over time between groups. Mechanical power ratio was the only physiological variable which remained consistently associated with ICU mortality throughout the study. The alteration in oxygenation, dead space, and mechanical power ratio should be assessed not at intensive care admission, but during the first days of mechanical ventilation to better predict outcome.


"Practice What You Teach" Public Health Nurses Promoting Healthy Lifestyles (PHeeL-PHiNe): Program Evaluation.
Healthy lifestyle programs are essential for meeting the challenge of noncommunicable diseases. The Public Health Nurses Promoting Healthy Lifestyles (PHeeL-PHiNe) program engaged nurses from family health clinics in Jerusalem District and included physical activity, healthy nutrition, and motivational skills. Questionnaires were completed at baseline, postintervention, and at 18 months. Results showed a marked effect on health practices. The proportion of nurses consuming a balanced diet and the use of food labels significantly increased and were maintained over time. Short-term improvements in physical activity were also observed. Nurses who practiced a healthy lifestyle were significantly more likely to provide guidance and counseling to families on healthy behaviors.


[Plasmapheresis as an experimental model for studies on the extracellular distribution of enzymes. Distribution and transport of cell enzymes within the extracellular space. IV (author's transl)].
The rate of distribution of cell enzymes between the intravascular and extravascular space was studied, following a sudden decrease of enzyme activities in plasma. This rapid decrease of enzyme activities was achieved in rats by a rapid exchange of the blood with a twofold volume of a suspension of homologous erythrocytes in isoosmolar bovine serum albumin solution. After this plasmapheresis, the activities of seven cell enzymes in the plasma were decreased to 14 to 22% of their original values. The subsequent increase in activities showed different kinetics, depending on the enzyme. After 120 min, creatine kinase had reached the starting activity; malate dehydrogenase and aldolase reached their original activities after 180 min. Aspartate aminotransferase, glutamate dehydrogenase, alanine aminotransferase and pyruvate kinase increased more slowly and they had still not reached their starting values after 240 min. Repetition of the plasmapheresis after 90 min had no obvious effect on the kinetics of the subsequent activity increase. During the first minutes after plasmapheresis the adjustment of the activity equilibrium between the interstitial and the intravascular compartments depends mainly on the capillary permeability. It is therefore possible to determine half-life constants for the distribution of enzymes within the extracellular space. The constants for malate dehydrogenase and aldolase are almost identical with those determined by intravenous injection, whereas there are discrepancies in the constants for the remaining enzymes. The constants for pyruvate kinase and glutamate dehydrogenase are significantly lower, while those for aspartate aminotransferase, alanine aminotransferase and creatine kinase are significantly higher, than those determined after intravenous injection. Possible reasons for these differences are disucssed.


Alpha-Actinin-4 is a Possible Target Protein for Aristolochic Acid I in Human Kidney Cells In Vitro.
Aristolochic acid I (AA-I) is a strong nephrotoxin, carcinogen, and mutagen found in plants such as the Aristolochia species. The mechanisms underlying AA-I toxicity in the kidneys are poorly understood. In this study, we aimed to gain insight into the mechanism of AA-I nephrotoxicity by analyzing the uptake, subcellular distribution, and intracellular targets of AA-I in the human kidney cell line HK-2 using immunocytochemistry, immunoprecipitation, and LC-MS/MS. In HK-2 cells incubated with 20[Formula: see text][Formula: see text]g/mL AA-I for different periods of time (up to 12[Formula: see text]h), AA-I was detected by a specific monoclonal antibody (MAb) against AA-I, both in the cytoplasm and nuclei. Nuclear localization depended on the exposure time. A protein with the molecular weight of 100 kDa was immunoprecipitated with the anti-AA-I MAb from the AA-I-treated cell lysates and was identified by LC-MS/MS as [Formula: see text]-actinin-4 after digestion of the protein, and was confirmed by immunoblotting with a specific anti-[Formula: see text]-actinin-4 MAb. This evidence shows, for the first time, that [Formula: see text]-actinin-4 is a protein targeted by AA-I in kidney cells. Our findings strongly suggest an association between [Formula: see text]-actinin-4 and AA-I nephrotoxic activity.


Prioritizing progressive MS rehabilitation research: A call from the International Progressive MS Alliance.
People with multiple sclerosis (MS) experience myriad symptoms that negatively affect their quality of life. Despite significant progress in rehabilitation strategies for people living with relapsing-remitting MS (RRMS), the development of similar strategies for people with progressive MS has received little attention. To highlight key symptoms of importance to people with progressive MS and stimulate the design and implementation of high-quality studies focused on symptom management and rehabilitation. A group of international research experts, representatives from industry, and people affected by progressive MS was convened by the International Progressive MS Alliance to devise research priorities for addressing symptoms in progressive MS. Based on information from the MS community, we outline a rationale for highlighting four symptoms of particular interest: fatigue, mobility and upper extremity impairment, pain, and cognitive impairment. Factors such as depression, resilience, comorbidities, and psychosocial support are described, as they affect treatment efficacy. This coordinated call to action-to the research community to prioritize investigation of effective symptom management strategies, and to funders to support them-is an important step in addressing gaps in rehabilitation research for people affected by progressive MS.


Gender-dependent levels of hyaluronic acid in cerebrospinal fluid of patients with neurodegenerative dementia.
Numerous reports over the years have described neuroinflammatory events and vascular changes in neurodegenerative diseases such as Alzheimers disease (AD) and Dementia with Lewy bodies (DLB). Interestingly, recent reports from other research areas suggest that inflammatory and vascular processes are influenced by gender. These findings are intriguing from the perspective that women show a higher incidence of AD and warrant investigations on how gender influences various processes in neurodegenerative dementia. In the current study we measured the cerebrospinal fluid (CSF) and plasma concentrations of hyaluroinic acid (HA), an adhesionmolecule known to regulate both vascular and inflammatory processes, in AD and DLB patients as well as in healthy elders. Our analysis showed that male AD and DLB patients had almost double the amount of HA compared to female patients whereas no gender differences were observed in the controls. Furthermore, we found that CSF levels of HA in foremost female AD patients correlated with various AD related biomarkers. Correlations between HA levels and markers of inflammation and vascular changes were only detected in female AD patients but in both male and female DLB patients. We conclude that HA may be linked to several pathological events present in AD, as reflected in CSF protein concentrations. The HA profile in CSF, but not in plasma, and associations to other markers appear to be gender-dependent which should be taken into account in clinical examinations and future biomarker studies.


Antiviral activity of monoclonal antibodies specific for the internal proteins N and NS of rabies virus.
Monoclonal antibodies (MAbs) specific for the rabies virus nucleoprotein (N protein) and non-structural (NS) protein of the nucleocapsid were introduced into adherent cells (fibroblasts and neuroblastoma) by the scrape-loading technique. After the cells had reattached to the substrate, they were infected with rabies virus. Inhibition of infection was monitored by measuring the intracytoplasmic viral nucleocapsid accumulation with an enzyme immunoassay using anti-N protein rabbit serum and by measuring the release of infectious virus with the plaquing system. Seventeen MAbs defining the three independent antigenic sites on the N protein were able to decrease nucleocapsid accumulation and the release of infectious virus. The MAbs describing the two antigenic sites on NS protein also had an antiviral effect on ERA virus. When an anti-N MAb (0.5 ng per cell) was introduced into CVS-infected cells, virus inhibition was complete if the anti-N MAb was introduced between 0 and 5 h post-infection and ineffective beyond 9 h post-infection. The inhibition was dose-dependent. The MAbs could block virus multiplication either by 'neutralizing' newly translated N and NS proteins or by impairing the initial transcription of the genome.


[Clinical features and outcome of treatment for novel coronavirus pneumonia: a meta-analysis].
Objective: To investigate the clinical features and outcome of treatment for novel coronavirus pneumonia. Methods: Literature on novel coronavirus pneumonia was retrieved from PubMed and EMBASE databases. The relevant data was extracted and a meta-analysis was performed using StatsDirect statistical software V.2.8.0 to calculate the combined odds ratio. Results: Seven studies were included, consisting of 1594 cases. The meta-analysis result showed that the most common clinical symptoms of the novel coronavirus pneumonia were fever (91.6%) and cough (64.5%), followed by dyspnea (32.8%) and sputum (28.1%). Headache (10.5%), sore throat (11.2%), hemoptysis (3.2%), diarrhea (6.6%) and the other symptoms were relatively rare. Aspartate aminotransferase (29%), alanine transaminase (22.7%), and total bilirubin (11.7%) levels were elevated, except for serum albumin levels (80.4%). The common therapeutic agents used were antibiotics (87.7%), antiviral drugs (75.5%), and glucocorticoids (26.6%), while antifungal agents (7.7%) were used in few. Mechanical ventilation (13.4%), extracorporeal membrane oxygenation (1.9%), and continuous renal replacement therapy (3.8%) were used in severe cases. The rate of mortality in hospital was 7.7%, respectively. Heterogeneity between studies was significant; however, subgroup and sensitivity analysis had failed to identify clear sources of heterogeneity. Conclusion: Fever, cough and liver dysfunction are the main clinical manifestations of this disease and the mortality rate is low.


Nanoscale segregation of actin nucleation and elongation factors determines dendritic spine protrusion.
Actin dynamics drive morphological remodeling of neuronal dendritic spines and changes in synaptic transmission. Yet, the spatiotemporal coordination of actin regulators in spines is unknown. Using single protein tracking and super-resolution imaging, we revealed the nanoscale organization and dynamics of branched F-actin regulators in spines. Branched F-actin nucleation occurs at the PSD vicinity, while elongation occurs at the tip of finger-like protrusions. This spatial segregation differs from lamellipodia where both branched F-actin nucleation and elongation occur at protrusion tips. The PSD is a persistent confinement zone for IRSp53 and the WAVE complex, an activator of the Arp2/3 complex. In contrast, filament elongators like VASP and formin-like protein-2 move outwards from the PSD with protrusion tips. Accordingly, Arp2/3 complexes associated with F-actin are immobile and surround the PSD. Arp2/3 and Rac1 GTPase converge to the PSD, respectively, by cytosolic and free-diffusion on the membrane. Enhanced Rac1 activation and Shank3 over-expression, both associated with spine enlargement, induce delocalization of the WAVE complex from the PSD. Thus, the specific localization of branched F-actin regulators in spines might be reorganized during spine morphological remodeling often associated with synaptic plasticity.


Immunoregulation through membrane proteins modified by reducing conditions induced by immune reactions.
Selected disulfide bonds in membrane proteins are labile and are thus susceptible to changes in redox potential and/or the presence of thiol isomerase enzymes. Modification of these disulfide bonds can lead to conformational changes of the protein that in turn may alter protein activity and function. This occurs in the entry of several enveloped viruses into their host cells, e.g. HIV, hepatitis C virus and Newcastle disease virus. Labile disulfide bonds are also important in platelet activation, cytokine signalling and in a variety of diseases including cancer and arthritis. In this review we will concentrate on recent advances in understanding the conditions that lead to disulfide bond reduction in membrane proteins and their effects in regulating immune function.


Implantable Self-Reporting Stents for Detecting In-Stent Restenosis and Cardiac Functional Dynamics.
Despite the increasing number of stents implanted each year worldwide, patients remain at high risk for developing in-stent restenosis. Various self-reporting stents have been developed to address this challenge, but their practical utility has been limited by low sensitivity and limited data collection. Herein, we propose a next-generation self-reporting stent that can monitor blood pressure and blood flow inside the blood arteries. This proposed self-reporting stent utilizes a larger inductor coil encapsulated on the entire surface of the stent strut, resulting in a 2-fold increase in the sensing resolution and coupling distance between the sensor and external antenna. The dual-pressure sensors enable the detection of blood flow in situ. The feasibility of the proposed self-reporting stent is successfully demonstrated through in vivo analysis in rats, verifying its biocompatibility and multifunctional utilities. This multifunctional self-reporting stent has the potential to greatly improve cardiovascular care by providing real-time monitoring and unprecedented insight into the functional dynamics of the heart.


Screening for GNAS genetic and epigenetic alterations in progressive osseous heteroplasia: first Italian series.
Progressive osseous heteroplasia (POH) is a rare autosomal dominant disorder of mesenchymal differentiation characterized by progressive heterotopic ossification (HO) of dermis, deep connective tissues and skeletal muscle. Usually, initial bone formation occurs during infancy as primary osteoma cutis (OC) then progressively extending into deep connective tissues and skeletal muscle over childhood. Most cases of POH are caused by paternally inherited inactivating mutations of GNAS gene. Maternally inherited mutations as well as epigenetic defects of the same gene lead to pseudohypoparathyroidism (PHP) and Albright's hereditary osteodystrophy (AHO). During the last decade, some reports documented the existence of patients with POH showing additional features characteristic of AHO such as short stature and brachydactyly, previously thought to occur only in other GNAS-associated disorders. Thus, POH can now be considered as part of a wide spectrum of ectopic bone formation disorders caused by inactivating GNAS mutations. Here, we report genetic and epigenetic analyses of GNAS locus in 10 patients affected with POH or primary OC, further expanding the spectrum of mutations associated with this rare disease and indicating that, unlike PHP, methylation alterations at the same locus are absent or uncommon in this disorder.


Machine-learning approaches for classifying haplogroup from Y chromosome STR data.
Genetic variation on the non-recombining portion of the Y chromosome contains information about the ancestry of male lineages. Because of their low rate of mutation, single nucleotide polymorphisms (SNPs) are the markers of choice for unambiguously classifying Y chromosomes into related sets of lineages known as haplogroups, which tend to show geographic structure in many parts of the world. However, performing the large number of SNP genotyping tests needed to properly infer haplogroup status is expensive and time consuming. A novel alternative for assigning a sampled Y chromosome to a haplogroup is presented here. We show that by applying modern machine-learning algorithms we can infer with high accuracy the proper Y chromosome haplogroup of a sample by scoring a relatively small number of Y-linked short tandem repeats (STRs). Learning is based on a diverse ground-truth data set comprising pairs of SNP test results (haplogroup) and corresponding STR scores. We apply several independent machine-learning methods in tandem to learn formal classification functions. The result is an integrated high-throughput analysis system that automatically classifies large numbers of samples into haplogroups in a cost-effective and accurate manner.


Brain-derived neurotrophic factor Val66Met polymorphism and early life adversity affect hippocampal volume.
The interaction between adverse life events during childhood and genetic factors is associated with a higher risk to develop major depressive disorder (MDD). One of the polymorphisms found to be associated with MDD is the Val66MET polymorphism of brain-derived neurotrophic factor (BDNF). The aim of our two-center study was to determine how the BDNF Val66Met polymorphism and childhood adversity affect the volumetric measures of the hippocampus in healthy individuals and people with MDD. In this two-center study, 62 adult patients with MDD and 71 healthy matched controls underwent high-resolution magnetic resonance imaging. We used manual tracing of the bilateral hippocampal structure with help of the software BRAINS2, assessed childhood adversity using the Childhood Trauma Questionnaire and genotyped Val66Met BDNF SNP (rs6265). MDD patients had smaller hippocampal volumes, both in the left and right hemispheres (F = 5.4, P = 0.022). We also found a significant interaction between BDNF allele and history of childhood adversity (F = 6.1, P = 0.015): Met allele carriers in our samples showed significantly smaller hippocampal volumes when they did have a history of childhood adversity, both in patients and controls. Our results highlight how relevant stress-gene interactions are for hippocampal volume reductions. Subjects exposed to early life adversity developed smaller hippocampal volumes when they carry the Met-allele of the BDNF polymorphism.


p.F508del in a heterogeneous cystic fibrosis population from Minas Gerais, Brazil.
Cystic fibrosis (CF) is the most common autosomal recessive disease of the Caucasian population. Among the various CF mutations, p.F508del is the most frequent, accounting for two-thirds of the global CF chromosomes, although showing great variability among populations. We have studied 115 unrelated CF patients from a mixed population of Minas Gerais (Brazil). To evaluate part of the DNA sequence of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, blood DNA was obtained and PCR was performed using two pairs of primers that anneal to exons 10 and 24 of the CFTR gene. The PCR product was then submitted to automatic sequencing using the ABI PRISM 310 Genetic Analyzer. The p.F508del mutation was found in 50 (21.7%) of 230 unrelated CF alleles. Fifteen (13.0%) patients were homozygous for this mutation, while 20 (17.4%) were heterozygous; the remaining 80 (69.6%) patients did not carry the p.F508del mutation. Exon 24 sequence had no change in 75 (65.2%) patients, 21 (18.3%) had the sequence variation 4521G/A, 11 (9.6%) had a not yet described sequence variation 4407T/A and 8 (7.0%) patients had both sequence variations (4521G/A and 4407T/A). The polymorphism 4407T/A results in an amino acid modification from aspartic acid to glutamic acid, which will probably have no function effect in CFTR. This low p.F508del prevalence can be due to the variable ethnic origin of this population from Minas Gerais, which may have a high diversity of CF rare mutations.


Our national nutrient reduction needs: Applying a conservation prioritization framework to US agricultural lands.
Targeted conservation approaches seek to focus resources on areas where they can deliver the greatest benefits and are recognized as key to reducing nonpoint source nutrients from agricultural landscapes into sensitive receiving waters. Moreover, there is growing recognition of the importance and complementarity of in-field and edge-of-field conservation for reaching nutrient reduction goals. Here we provide a generic prioritization that can help with spatial targeting and applied it across the conterminous US (CONUS). The prioritization begins with identifying areas with high agricultural nutrient surplus, i.e., where the most nitrogen (N) and/or phosphorus (P) inputs are left on the landscape after crop harvest. Subwatersheds with high surplus included 52% and 50% of CONUS subwatersheds for N and P, respectively, and were located predominantly in the Midwest for N, in the South for P, and in California for both N and P. Then we identified the most suitable conservation strategies using a hierarchy of metrics including nutrient use efficiency (proportion of new nutrient inputs removed by crop harvest), tile drainage, existing buffers for agricultural run-off, and wetland restoration potential. In-field nutrient input reduction emerged as a priority because nutrient use efficiency fell below a high but achievable goal of 0.7 (30% of nutrients applied are not utilized) in 45% and 44% of CONUS subwatersheds for N and P, respectively. In many parts of the southern and western US, in-field conservation (i.e., reducing inputs + preventing nutrients from leaving fields) alone was likely the optimal strategy as agriculture was already well-buffered. However, stacking in-field conservation with additional edge-of-field buffering would be important to conservation strategies in 35% and 29% of CONUS subwatersheds for N and P, respectively. Nutrient use efficiencies were often high enough in the Midwest that proposed strategies focused more on preventing nutrients from leaving fields, managing tile effluent, and buffering agricultural fields. Almost all major river basins would benefit from a variety of nutrient reduction conservation strategies, underscoring the potential of targeted approaches to help limit excess nutrients in surface and ground waters.


Ballistic PbTe Nanowire Devices.
Disorder is the primary obstacle in the current Majorana nanowire experiments. Reducing disorder or achieving ballistic transport is thus of paramount importance. In clean and ballistic nanowire devices, quantized conductance is expected, with plateau quality serving as a benchmark for disorder assessment. Here, we introduce ballistic PbTe nanowire devices grown by using the selective-area-growth (SAG) technique. Quantized conductance plateaus in units of 2e2/h are observed at zero magnetic field. This observation represents an advancement in diminishing disorder within SAG nanowires as most of the previously studied SAG nanowires (InSb or InAs) have not exhibited zero-field ballistic transport. Notably, the plateau values indicate that the ubiquitous valley degeneracy in PbTe is lifted in nanowire devices. This degeneracy lifting addresses an additional concern in the pursuit of Majorana realization. Moreover, these ballistic PbTe nanowires may enable the search for clean signatures of the spin-orbit helical gap in future devices.


Characterization of a recombinant transferrin-binding protein A (TbpA) fragment from Haemophilus parasuis serovar 5.
Haemophilus parasuis, the etiological agent of Glässer's disease in pigs, possesses iron acquisition pathways mediated by a surface receptor that specifically bind porcine transferrin. This receptor is composed of transferrin-binding protein A (TbpA) and TbpB. As it has been reported for other gram-negative organisms, H. parasuis TbpA could be useful as a candidate target for H. parasuis vaccination. In this study, a 600-bp tbpA fragment of the gene encoding TbpA from H. parasuis serovar 5, the Nagasaki strain, was amplified by PCR and cloned into a pBAD/Thio-TOPO expression vector, generating the pBAD-Thio-TbpA-V5-His (TbpA-His) construction. Escherichia coli LMG194-competent cells were transformed with this construction, followed by the induction of protein expression with arabinose. A band (38.5 kDa) corresponding to a 200-amino acid recombinant TbpA (rTbpA) fragment was seen on the sodium dodecyl sulfate polyacrylamide gel electrophoresis and confirmed by immunoblotting. Polyclonal antibodies raised against this fragment were specific for H. parasuis and Actinobacillus pleuropneumoniae, reacted at the cell surface with H. parasuis, and a significant bactericidal activity was also detected. Therefore, this rTbpA fragment induces an immunological response and might be useful as an antigen for vaccination against Glässer's disease.


Combined effect of fungicide, herbicide and plant elicitor used in apple orchards on non-target epiphytic moss Hypnum cupressiforme.
Apple production is a dynamic agricultural system in which pesticides are applied recurrently to control pests and diseases in the orchards. Understanding the impact of such agents on non-target organisms is crucial to minimise unintended consequences while maintaining their use in crop protection. The aim was to test how fungicide, herbicide, elicitor, and their combinations affect the physiology of the epiphytic moss Hypnum cupressiforme that naturally occurs in orchards. Our results showed that both dodine and diflufenican applied separately had a strong negative effect on moss physiology reflected in significantly decreased photosynthetic pigment contents, maximum quantum yield of PSII photochemistry, cell membrane integrity and dehydrogenase activity, and increased membrane lipid peroxidation, which indicates a high physiological stress. Furthermore, the combined use of herbicide and fungicide resulted in further deterioration of the physiological condition compared to the effects of both agents used separately. In many cases, the application of chitosan together with a diflufenican or dodine resulted in a reduction of the negative effects triggered by these agents. The compensatory effect was particularly pronounced in maintaining a low level of cell membrane permeability. Consequently, it can be concluded that chitosan could have a protective function against cell membrane damage in non-target mosses.


Human cytomegalovirus and Epstein-Barr virus infection in inflammatory bowel disease: need for mucosal viral load measurement.
To evaluate the best diagnostic technique and risk factors of the human Cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) infection in inflammatory bowel disease (IBD). A cohort of 40 IBD patients (17 refractory) and 40 controls underwent peripheral blood and endoscopic colonic mucosal sample harvest. Viral infection was assessed by quantitative real-time polymerase chain reaction and immunohistochemistry, and correlations with clinical and endoscopic indexes of activity, and risk factors were investigated. All refractory patients carried detectable levels of HCMV and/or EBV mucosal load as compared to 13/23 (56.5%) non-refractory and 13/40 (32.5%) controls. The median DNA value was significantly higher in refractory (HCMV 286 and EBV 5.440 copies/10(5) cells) than in non-refractory (HCMV 0 and EBV 6 copies/10(5) cells; P < 0.05 and < 0.001) IBD patients and controls (HCMV and EBV 0 copies/10(5) cells; P < 0.001 for both). Refractory patients showed DNA peak values ≥ 10(3) copies/10(5) cells in diseased mucosa in comparison to non-diseased mucosa (P < 0.0121 for HCMV and < 0.0004 for EBV), while non-refractory patients and controls invariably displayed levels below this threshold, thus allowing us to differentiate viral colitis from mucosal infection. Moreover, the mucosal load positively correlated with the values found in the peripheral blood, whilst no correlation with the number of positive cells at immunohistochemistry was found. Steroid use was identified as a significant risk factor for both HCMV (P = 0.018) and EBV (P = 0.002) colitis. Finally, a course of specific antiviral therapy with ganciclovir was successful in all refractory patients with HCMV colitis, whilst refractory patients with EBV colitis did not show any improvement despite steroid tapering and discontinuation of the other medications. Viral colitis appeared to contribute to mucosal lesions in refractory IBD, and its correct diagnosis and management require quantitative real-time polymerase chain reaction assay of mucosal specimens.