Clinical development of gene therapy: results and lessons from recent successes.
Therapeutic gene transfer holds the promise of providing lasting therapies and even cures for diseases that were previously untreatable or for which only temporary or suboptimal treatments were available. For some time, clinical gene therapy was characterized by some impressive but rare examples of successes and also several setbacks. However, effective and long-lasting treatments are now being reported from gene therapy trials at an increasing pace. Positive outcomes have been documented for a wide range of genetic diseases (including hematological, immunological, ocular, and neurodegenerative and metabolic disorders) and several types of cancer. Examples include restoration of vision in blind patients, eradication of blood cancers for which all other treatments had failed, correction of hemoglobinopathies and coagulation factor deficiencies, and restoration of the immune system in children born with primary immune deficiency. To date, about 2,000 clinical trials for various diseases have occurred or are in progress, and many more are in the pipeline. Multiple clinical studies reported successful treatments of pediatric patients. Design of gene therapy vectors and their clinical development are advancing rapidly. This article reviews some of the major successes in clinical gene therapy of recent years.


Impact of the nurse-related information through social media use on undergraduate nursing students' professional identity in nursing: A mixed-methods study.
To understand the impact of nurse-related information, through social media usage on undergraduate nursing students' professional identity. Previous research has addressed professional knowledge and practice that affect the development of undergraduate nursing students' professional identities. As an increasingly crucial factor in the internet age, social media needs to be explored in-depth for its influence on the professional identity in nursing. Sequential explanatory mixed method was used in this study. In this quantitative study, participants (N = 698) completed an e-questionnaire survey between June and July of 2020. The relationship between main variables was analyzed by linear regression using SPSS 20.0. Then, face-to-face semi-structured and audio-recorded interviews were carried out among participants (N = 16) between January and March of 2021. The quantitative findings indicated that the mean score of professional identity was 93.07 ± 11.96 in participants. The results of the linear regression showed that the year in school, whether nursing was the first choice, online time per day, frequency of reading official accounts related nursing on WeChat and appraisal of the online nurse-related information and so on explained 49.0 % of the variance in professional identity. Four major themes emerged from the qualitative study: 1) increased concern; 2) few and non-professional information; 3) encouraged by positive information; 4) limited impact of negative information. Nurse-related information on social media has the potential to have an impact on undergraduate nursing students' professional identity. Nursing educators should guide undergraduate nursing students to use social media appropriately and assist them in developing their professional identity with it.


Evaluation of the plasma microRNA levels in stage 3 premature retinopathy with plus disease: preliminary study.
PurposeIn the present study, we aimed to investigate the changes in plasma microRNA (miRNA) levels in premature infants diagnosed with premature retinopathy (ROP).Patients and methodsIn order to investigate the relationship of miRNAs with ROP, 13 premature infants admitted to Mersin University, Medical School, Department of Ophthalmology and diagnosed with ROP stage 3 with plus disease between January 2014-January 2015 were included in the study. Control group consisted of 15 premature infants with no ROP. The plasma miRNA levels were evaluated using high-throughput quantitative real-time PCR.ResultsThe results of study demonstrated that the expression level of miR-23a and miR-200b-3p was significantly upregulated in patients with ROP when compared with the control group (P<0.05). The expression level of miR-27b-3p and miR-214-3p was significantly downregulated in patients (P<0.05). In addition, expression of 17 miRNA (miR-410-3p, miR-17-5p, miR-451a, miR-31-5p, miR-132-3p, miR-183-5p, miR-184, miR-222-3p, miR-296-5p, miR-200a-3p, miR-328-3p,miR-96-5p, miR-199a-5p, miR-99a-5p, miR-106a-5p, miR-125b-5p, miR-155-5p) had upregulated or downregulated, but not statistically significantly different when compared with the control group.ConclusionsOur results suggest that plasma miRNA levels may alter in ROP and, some miRNAs might have an effect in the physiopathology of this disease. These molecules may have an important therapeutic role in patients who are unresponsive to anti-vascular endothelial growth factor therapy. However, further studies must be conducted for possible effects of miRNAs in vascular disorders of eye such as ROP. Moreover to define the relationship of these molecules with the disease more clearly, a multicenter study including more patients is necessary.


A reactive oxygen species burst causes haploid induction in maize.
Haploid induction (HI) is an important tool in crop breeding. Phospholipase A1 (ZmPLA1)/NOT LIKE DAD (NLD)/MATRILINEAL (MTL) is a key gene controlling HI in maize; however, the underlying molecular mechanism remains unclear. In this study, to dissect why loss of ZmPLA1 function could mediate HI we performed a comprehensive multiple omics analysis of zmpla1 mutant anthers by integrating transcriptome, metabolome, quantitative proteome, and protein modification data. Functional classes of significantly enriched or differentially abundant molecular entities were found to be associated with the oxidative stress response, suggesting that a reactive oxygen species (ROS) burst plays a critical role in HI. In support of this, we further discovered that a simple chemical treatment of pollen with ROS reagents could lead to HI. Moreover, we identified ZmPOD65, which encodes a sperm-specific peroxidase, as a new gene controlling HI. Taken together, our study revealed a likely mechanism of HI, discovered a new gene controlling HI, and created a new method for HI in maize, indicating the importance of ROS balance in maintaining normal reproduction and providing a potential route to accelerate crop breeding.


Effects of alteplase in the treatment of acute ischemic stroke.
For the last 15 years, alteplase (recombinant tissue plasminogen activator) has been used widely throughout the world for the treatment of acute ischemic stroke. Although considered to be safe and effective, like all drugs, alteplase has side effects. This retrospective cohort study was conducted in the intensive care unit of the department of internal medicine in a mid-size peripheral acute hospital in Germany. Patients with acute ischemic stroke who underwent alteplase-induced thrombolysis were investigated. Among the 1017 patients admitted for stroke investigation, 23 (2.26%) received thrombolytic therapy consisting of intravenous alteplase. Of these, six patients (26.09%) experienced complications, ie, four (17.39%) had intracerebral hemorrhage, one (4.35%) developed orolingual angioedema, and one (4.35%) had a hematoma on the right arm. After treatment with alteplase, two (33.33%) patients in the study group (n = 6) died because of intracerebral hemorrhage and one (16.67%) died because of aspiration pneumonia. One (5.88%) patient in the control group (n = 17) died of cerebral edema. The incidence of stroke and number of patients treated with alteplase in the examined hospital subunit has not increased in recent years. Also, in this study, no statistically significant difference was found in the incidence of various complications occurring during treatment for acute ischemic stroke with alteplase, but intracerebral hemorrhage was the most common complication.


[Monoclonal antibodies--new probes for diagnosis and therapy. Their use as an example of the micrometastasizing of solid tumors].
Monoclonal antibody CK2, recognizing component No. 18, appeared to be the most suitable reagent for the detection of epithelial tumor cells in bone marrow. Its specificity was confirmed in a double-marker staining procedure (combination of APAAP-technique and radioautography). CK2 positive cells were demonstrated not to reveal any cross-reactivity with an antibody directed against the "leucocyte common antigen". A significant correlation between the presence of epithelial tumor cells in bone marrow and certain conventional risk factors was found. A more detailed phenotypic characterisation could demonstrate the expression of proliferation associated antigens on these cells. Furthermore in an immunotherapeutic approach with monoclonal antibody 17-1A, labelling of the disseminated tumor cells in bone marrow after infusion of the antibody was shown.


Anti-Melanogenic Effects of Ethanol Extracts of the Leaves and Roots of Patrinia villosa (Thunb.) Juss through Their Inhibition of CREB and Induction of ERK and Autophagy.
Patrinia villosa (Thunb.) Juss is a traditional herb commonly used in East Asia including Korea, Japan, and China. It has been administered to reduce and treat inflammation in Donguibogam, Korea. The mechanism for its anti-inflammatory effects has already been reported. In this study, we confirmed the efficacy of Patrinia villosa (Thunb.) Juss ethanol extract (Pv-EE) for inducing autophagy and investigate its anti-melanogenic properties. Melanin secretion and content were investigated using cells from the melanoma cell line B16F10. Pv-EE inhibited melanin in melanogenesis induced by α-melanocyte-stimulating hormone (α-MSH). The mechanism of inhibition of Pv-EE was confirmed by suppressing the mRNA of microphthalmia-associated transcription factor (MITF), decreasing the phosphorylation level of CREB, and increasing the phosphorylation of ERK. Finally, it was confirmed that Pv-EE induces autophagy through the autophagy markers LC3B and p62, and that the anti-melanogenic effect of Pv-EE is inhibited by the autophagy inhibitor 3-methyl adenine (3-MA). These results suggest that Pv-EE may be used as a skin protectant due to its anti-melanin properties including autophagy.


Injuries of the cervical spine. Clinical and radiological results in 119 patients.
Seventy-one patients of 119 with injury of the cervical spine were re-examined an average of 5 years and 5 months after the accident. The mean age of the patients was 39 years. There were 91 male and 28 female patients. The majority of injuries (51) were sustained by car drivers and passengers involved in traffic accidents. More than one cervical vertebra was damaged in 36 patients. Neurological lesions were observed in 59 patients; tetraplegia was present in 19 of these. The average duration of hospital treatment was 37 days. Twenty-four patients were treated at the intensive care unit. Skull traction was used in 69 cases for an average of 38 days to bring about reduction and immobilization. Reduction and anterior spondylodesis by Cloward's technique was performed on 13 patients. Of the operated patients, 11 showed root symptoms or incomplete cord lesions. The 19 tetraplegics were all conservatively treated. Fourteen patients died of their cervical spine injuries and 7 remained permanently disabled. Half the patients recovered to a pain-free state and one third became asymptomatic. Of the radiological deformities observed on admission, lowering of the disc space increased during follow-up, and forward sliding, increase in interspinous distance and narrowing of the vertebral canal decreased. The clinical result was poorer if flexion deformity, compression, forward sliding, narrowing of the vertebral canal or lowering of the disc space were observed at the last radiological examination.


Synthesis, characterization and antibacterial activity of novel heterocycle, coumacine, and two of its derivatives.
Heterocyclic nucleus plays a fundamental role in the medicinal chemistry and serves as a key template for the development of various therapeutic agents including broad spectrum antibacterial drugs. In an effort to develop new antibacterial agents, a bicyclic twelve-membered heterocyclic nucleus derived from coumarin was prepared by an uncomplicated method. The rate of ring closure for this nucleus, which was given the name coumacine, in addition to two of its derivatives was monitored spectroscopically and this rate followed zero order kinetics. The chemical structures of the synthesized products were established by detecting their physicochemical properties and analyzing their IR, 1H NMR and 13C NMR spectra. The in vitro antibacterial activity of coumacines was evaluated via agar dilution method against different standard aerobic and anaerobic bacterial strains using ciprofloxacin and metronidazole as positive controls, respectively; the results indicated that coumacine I has an excellent broad spectrum antibacterial activity against the tested bacterial strains with percentage of growth inhibition approximating to those of positive controls.


Pain Characteristics, Psychosocial Wellbeing, and Sexual Wellbeing of Women Diagnosed With Provoked Vestibulodynia and a History of Sexual Abuse.
Provoked vestibulodynia (PVD) is a common chronic pain condition characterized by pain at the vulvar vestibule elicited by touch. Both PVD and sexual abuse lead to negative psychosocial and sexual consequences. However, little is known about the wellbeing of women with PVD and a history of sexual abuse. The aim of this study was to characterize a sample of women seeking treatment for PVD who have experienced sexual abuse. A total of 404 women diagnosed with PVD completed self-report questionnaires of PVD symptoms and psychosocial and sexual wellbeing before and after participating in a multidisciplinary PVD treatment program. History of sexual abuse was assessed via self-report, and women were dichotomized into groups. No significant differences were found on sociodemographic variables, baseline psychosocial or sexual functioning between women with and without a self-reported history of sexual abuse (n = 40 and n = 364, respectively). Significantly more women with a history of sexual abuse than without reported other comorbid chronic pain conditions and radiating PVD pain. History of sexual abuse did not affect improvements in sexual distress scores following multidisciplinary treatment for their PVD. Ten percent of women in our sample self-reported a history of sexual abuse, but the two groups did not differ significantly with respect to their baseline psychosocial or sexual functioning concerns, and both groups reported reductions in sexual distress following treatment for PVD. These findings indicate that a history of sexual abuse does not significantly affect the efficacy of multidisciplinary treatment approaches for PVD.


Effect of risk factors and antirheumatic drugs on the proliferation of aortic wall cells.
The proliferation of aortic smooth muscle cells (ASMC) of Wistar rats, impaired by risk factors such as arterial hypertension, diabetes mellitus, atherogenic diet and staphylolysin injections and of normal Wistar rats treated with antirheumatic drugs such as prednisolone and acetylsalicylic acid was investigated. The cells of these animals were cultivated, subcultivated, and in the 2nd subcultures the cell numbers/5 ml medium were counted by means of Coulter Counter, and the cells were incubated with [3H]thymidine and the percentage of labelling in 100 or 1000 counted cells was stated. The effect of risk factors such as LDL and staphylolysin and of antirheumatic drugs such as prednisolone, acetylsalicylic acid, D-penicillamine and chloroquine added to the 2nd subcultures of cultivated ASMC of normal minipigs was investigated by the same method. The proliferation of cultivated ASMC of rats impaired by risk factors was accelerated. The proliferation of cultivated ASMC of rats treated with antirheumatic drugs was inhibited. The proliferation of ASMC of minipigs in the 2nd subcultures was activated by addition of risk factors and inhibited by addition of antirheumatic drugs. Antirheumatic drugs given to the rats and added to the medium of the 2nd subcultures of ASMC of normal minipigs inhibit the acceleration of ASMC proliferation induced by simultaneously given risk factors. The proposal to augment up our arsenal of the hitherto existing preventive and therapeutical measures by the application of antirheumatic drugs based on the experimental models referred to is supported by the result of a limited prospective double-blind-study of a sample of 133 male patients after myocardial infarction. The most remarkable result that the acceleration of the ASMC proliferation, the real pathologic process of arteriosclerosis, is inhibited by the application of antirheumatic drugs, at exactly the same time as the acceleration of the fibroblast proliferation, the real pathologic process in rheumatic diseases--ASMC and fibroblast, both being mesenchymal cells--recommends the use of these drugs in the prevention and therapy of human arteriosclerosis. The surprising result of our in-vivo experiments, that the acceleration of the growth of the ASMC induced by risk factors and the inhibition of the growth induced by antirheumatic drugs persist in the subcultures, is explained by the "selection theory" that there are dissimilar kinds of ASMC in normal arteries and that they react differently.


Validating measures of third year medical students' use of interpreters by standardized patients and faculty observers.
Increasing prevalence of limited English proficiency patient encounters demands effective use of interpreters. Validated measures for this skill are needed. We describe the process of creating and validating two new measures for rating student skills for interpreter use. Encounters using standardized patients (SPs) and interpreters within a clinical practice examination (CPX) at one medical school. Students were assessed by SPs using the interpreter impact rating scale (IIRS) and the physician patient interaction (PPI) scale. A subset of 23 encounters was assessed by 4 faculty raters using the faculty observer rating scale (FORS). Internal consistency reliability was assessed by Cronbach's coefficient alpha (alpha). Interrater reliability of the FORS was examined by the intraclass correlation coefficient (ICC). The FORS and IIRS were compared and each was correlated with the PPI. Cronbach's alpha was 0.90 for the 7-item IIRS and 0.88 for the 11-item FORS. ICC among 4 faculty observers had a mean of 0.61 and median of 0.65 (0.20, 0.86). Skill measured by the IIRS did not significantly correlate with FORS but correlated with the PPI. We developed two measures with good internal reliability for use by SPs and faculty observers. More research is needed to clarify the reasons for the lack of concordance between these measures and which may be more valid for use as a summative assessment measure.


Improvement of soft ground using solidified coal ash and its effects on the marine environment.
This paper presents some research results pertaining to the application of solidified coal ash (SCA) for improving the stability of soft ground. The results pertain to the physical properties of SCA required for use as an alternative to sand in sand compaction piles (SCPs), and the assessment of possible environmental impacts resulting from construction of SCA piles in marine environments. The results of field tests indicate that the physical properties of SCA (permeability, internal friction angle, and grain-size distribution) are favorable for use in soil improvement applications. Also, the results show that SCA is sufficiently suitable as an alternative to sand in SCPs, although SCA piles cannot be compacted to the same extent as sand piles. Finally, test results showed no adverse environmental impacts on natural benthos resulting from placement of SCA piles in marine environments. Thus, the results of this study confirm that SCA is a viable alternative material to sand in SCPs that are used for ground improvement in marine environments, and that large quantities of SCA may be required for such applications resulting in an alternative use for an otherwise waste material.


Anlotinib optimizes anti-tumor innate immunity to potentiate the therapeutic effect of PD-1 blockade in lung cancer.
Many anti-angiogenic agents have the potential to modulate the tumor microenvironment and improve immunotherapy. Anlotinib has demonstrated anti-tumor efficacy in non-small cell lung cancer (NSCLC) in third-line clinical trials. However, its roles in immune regulation and potentially synergistic anti-tumor effect in combination with immune checkpoint inhibition remain unclear. Here, based on a syngeneic lung cancer mouse model, the intratumoral immunological changes post-anlotinib treatment in the model were assessed. Furthermore, it was tested whether anlotinib could enhance the anti-tumor effect of αPD-1 in vivo. This study shows that anlotinib increased infiltration of the innate immune cells, including natural killer (NK) cells, and antigen-presenting cells (APC), which include M1-like tumor-associated macrophages (TAM) and dendritic cells (DC), whereas the percentage of M2-like TAM was dramatically reduced. Subsequently, when combined with PD-1/PD-L1 (programmed cell death 1/PD-1 ligand 1) blockade, anlotinib conferred significantly synergistic therapeutic benefits. Overall, these findings describe a role for anlotinib in the innate immune cells in the tumor microenvironment and a potentially synergistic anti-tumor combination with immune checkpoint inhibition.


Efficient immobilized phospholipase A1 on Mo-basing nanomaterials for enzymatic degumming.
Six kinds of Mo-basing nanomaterials (MoO3 , MoO3 @Ru, Mo-PDA, MoPC , MoP, CNT@MoS2 ) were successfully synthesized, which were employed as carriers to immobilize phospholipase A1 (PLA1) for the hydrolysis of phospholipids (PLs). PLA1 was immobilized by a simple adsorption-precipitation-cross-linking to form an "enzyme net" covering on nanoparticles. The greatest advantage of these nanoparticles was their strong hydrophilic surface. It not only permitted their dispersion in the aqueous phase, but also showed the strong affinity for PLs in the organic phase, because amphiphilic PLs had the polar head group and higher hydrophilicity than other oils components. Michaelis-Menten analysis revealed that higher catalytic activity and enzyme-substrate affinity were observed in several immobilized PLA1 than its free form. MoO3 was confirmed to be the best candidate for carrier. The highest specific activity of MoO3 -immobilized PLA1 reached 43.1 U/mg, which was about 1.8 times higher than that of free PLA1 (24.4 U/mg). In addition, the stability and recycling were also enhanced. The robust immobilized PLA1 was prepared in this work, showing great potential for the enzymatic degumming.


Two-Dimensional Fluorinated Covalent Organic Frameworks with Tunable Hydrophobicity for Ultrafast Oil-Water Separation.
Covalent organic frameworks (COFs) hold great potentials for addressing the challenge of highly efficient oil-water separation, but they are restricted by the poor wettability and processability as crystalline membranes. Here we report the design and synthesis of two-dimensional (2D) robust COFs with controllable hydrophobicity and processability, allowing the COF layers to be directly grown on the support surfaces. Three 2D COFs with AA or ABC stacking are prepared by condensation of triamines with fluorine and/or isopropyl groups and perfluorodialdehyde. They all show excellent tolerance to water, acid, and base, with water contact angles (CA) of 111.5-145.8°. The two COFs with isopropyl and fluorine mixtures can grow as a coating on a stainless-steel net (SSN) substrate, whereas the one with only fluorine substituents cannot. The superhydrophobic COF@SSN coating with water CA of up to 150.1° displays high water-resistance and self-cleaning properties, enabling high oil-water separation performances with an efficiency of over 99.5 % and a permeation flux of 2.84×105 L m-2 h-1 , which are among the highest values reported for state-of-the-art membranes.


Shifting the mindset culture to address global educational disparities.
Educational outcomes remain highly unequal within and across nations. Students' mindsets-their beliefs about whether intellectual abilities can be developed-have been identified as a potential lever for making adolescents' academic outcomes more equitable. Recent research, however, suggests that intervention programs aimed at changing students' mindsets should be supplemented by programs aimed at the changing the mindset culture, which is defined as the shared set of beliefs about learning in a school or classroom. This paper reviews the theoretical and empirical origin of the mindset culture and examines its potential to reduce group-based inequalities in education. In particular, experiments have identified two broad ways the mindset culture is communicated by teachers: via informal messages about growth (e.g., that all students will be helped to learn and succeed), and formal opportunities to improve (e.g., learning-focused grading policies and opportunities to revise and earn credit). New field experiments, applying techniques from behavioral science, have also revealed effective ways to influence teachers' culture-creating behaviors. This paper describes recent breakthroughs in the U.S. educational context and discusses how lessons from these studies might be applied in future, global collaborations with researchers and practitioners.


Gender differences in rotation of the shank during single-legged drop landing and its relation to rotational muscle strength of the knee.
Increased shank rotation during landing has been considered to be one of the factors for noncontact anterior cruciate ligament injuries in female athletes. There have been no known gender differences in rotational knee muscle strength, which is expected to inhibit exaggerated shank rotation. Women have less knee external rotator strength than do men. Lower external rotator strength is associated with increased internal shank rotation at the time of landing. Controlled laboratory study. One hundred sixty-nine healthy young subjects (81 female and 88 male; age, 17.0 +/- 1.0 years) volunteered to participate in this study. The subjects performed single-legged drop landings from a 20-cm height. Femoral and shank kinematics were measured using a 3D optoelectronic tracking system during the drop landings, and then the joint angles around the knee (flexion/extension, valgus/varus, and internal/external rotation) were calculated. The maximal isometric rotational muscle strength of the knee was measured at 30 degrees of knee flexion in a supine position using a dynamometer. The female subjects had significantly less external shank rotation strength than did the male subjects (P < .001). Female subjects also exhibited significantly greater peak shank internal rotation angles than did males during landing (P < .05). Moderate but significant association was found between the maximum shank external rotation strength and the peak shank internal rotation angle during landing (r = -0.322, P < .01). Female subjects tended to have poor shank external rotator strength. This may lead to large shank internal rotation movement during the single-legged drop landing. Improving strength training of the external rotator muscle may help decrease the rates of anterior cruciate ligament injury in female athletes.


N,N-di(2-pyridylmethyl)amino-modified porphyrinato zinc complexes. The "ON-OFF" fluorescence sensor for Fe3+.
Two porphyrinato-zinc derivatives modified with two and four N,N-di(2-pyridylmethyl)amino groups, namely Zn-Porphyrin-2-DPA (1) and Zn-Porphyrin-4-DPA (2), have been designed, synthesized, and characterized. Systemic studies over a series of metal ions reveal that binding with Fe(3+) for the former compound induces the decrease of fluorescence emission intensity by 37.2%, while for the latter one results in a more remarkable decrease in the fluorescence emission intensity by 90.3%, rendering compound 2 the first example of DPA-modified porphyrin fluorescence ON-OFF sensor for Fe(3+).


Structural analysis of nucleosomal barrier to transcription.
Thousands of human and Drosophila genes are regulated at the level of transcript elongation and nucleosomes are likely targets for this regulation. However, the molecular mechanisms of formation of the nucleosomal barrier to transcribing RNA polymerase II (Pol II) and nucleosome survival during/after transcription remain unknown. Here we show that both DNA-histone interactions and Pol II backtracking contribute to formation of the barrier and that nucleosome survival during transcription likely occurs through allosterically stabilized histone-histone interactions. Structural analysis indicates that after Pol II encounters the barrier, the enzyme backtracks and nucleosomal DNA recoils on the octamer, locking Pol II in the arrested state. DNA is displaced from one of the H2A/H2B dimers that remains associated with the octamer. The data reveal the importance of intranucleosomal DNA-protein and protein-protein interactions during conformational changes in the nucleosome structure on transcription. Mechanisms of nucleosomal barrier formation and nucleosome survival during transcription are proposed.


Classical and molecular cytogenetics of the zebrafish, Danio rerio (Cyprinidae, Cypriniformes): an overview.
The zebrafish, Danio rerio, has recently become the model system for the genetic analysis of vertebrate development. This paper reviews the advances in zebrafish cytogenetics, obtained through classical and molecular techniques, which will lead to the assignment of specific linkage groups to specific chromosome pairs in the zebrafish genome project. Several chromosome pairs of the 50-chromosome karyotype of D. rerio were differentially stained by classical staining techniques and additional information has been obtained by molecular cytogenetics. Indeed, the analysis of constitutive heterochromatin by C-banding and base-specific fluorochrome staining had suggested a differential composition of peri- and paracentromeric constitutive heterochromatin. The chromosome mapping of distinct AT- and GC-rich zebrafish satellite DNAs by means of PRINS (Primed in situ) and multicolor FISH (Fluorescence in situ Hybridization) has confirmed this hypothesis, which therefore provided the chromosome localization of 10% of the zebrafish genome. The analysis of nucleolus organizer regions (NORs) by silver staining and by FISH with 18S rDNA has also revealed the existence of variable and inactive NORs, in addition to those on the terminal regions of the long arms of the three NOR-bearing chromosome pairs. Other multicopy genes, such as minor ribosomal genes, or multicopy repeats, such as telomere specific sequences, have now been mapped on zebrafish chromosomes. The latest advancement in zebrafish molecular cytogenetics is the chromosome mapping of single locus genes. Single-copy genes from each of the 25 genetic linkage groups are now being mapped on zebrafish chromosomes by using PAC clones.


Purification and cloning of interferon alpha.
Interferon alpha (IFN-alpha) was approved by the US Food and Drug Administration on June 5, 1986 and paved the way for development of many other cytokines and growth factors. Nevertheless, we have barely touched the surface of understanding the multitude of human IFNs. This paper reviews the history of the purification of human leukocyte IFN, the cloning of the IFN-alphas, and the current state of knowledge of human interferon alpha genes and proteins.


Small substrate transport and mechanism of a molybdate ATP binding cassette transporter in a lipid environment.
Embedded in the plasma membrane of all bacteria, ATP binding cassette (ABC) importers facilitate the uptake of several vital nutrients and cofactors. The ABC transporter, MolBC-A, imports molybdate by passing substrate from the binding protein MolA to a membrane-spanning translocation pathway of MolB. To understand the mechanism of transport in the biological membrane as a whole, the effects of the lipid bilayer on transport needed to be addressed. Continuous wave-electron paramagnetic resonance and in vivo molybdate uptake studies were used to test the impact of the lipid environment on the mechanism and function of MolBC-A. Working with the bacterium Haemophilus influenzae, we found that MolBC-A functions as a low affinity molybdate transporter in its native environment. In periods of high extracellular molybdate concentration, H. influenzae makes use of parallel molybdate transport systems (MolBC-A and ModBC-A) to take up a greater amount of molybdate than a strain with ModBC-A alone. In addition, the movement of the translocation pathway in response to nucleotide binding and hydrolysis in a lipid environment is conserved when compared with in-detergent analysis. However, electron paramagnetic resonance spectroscopy indicates that a lipid environment restricts the flexibility of the MolBC translocation pathway. By combining continuous wave-electron paramagnetic resonance spectroscopy and substrate uptake studies, we reveal details of molybdate transport and the logistics of uptake systems that employ multiple transporters for the same substrate, offering insight into the mechanisms of nutrient uptake in bacteria.


Allogeneic CD34-selected stem cell boost as salvage treatment of life-threatening infection and severe cytopenias after CAR-T cell therapy.
A variable incidence of profound cytopenia has been described in patients receiving chimeric antigen receptor T-cell (CAR-T) therapy for relapsed or refractory (R/R) B-cell acute lymphoblastic leukemia (ALL). This complication leads to severe infection in some cases, especially those who present additional risk factors including prior hematopoietic stem cell transplantation (HSCT). We report a case of breakthrough invasive fungal infection in a patient with prolonged neutropenia after CAR-T cell therapy administered for relapsed B-cell ALL after allogeneic haploidentical HSCT. After disease progression was discarded, therapy with antifungal agents, G-CSF and thrombopoietin analogue was started. However, no sign of haematological recovery or infection improvement was observed. A fresh mobilized selected CD34-stem cell boost from her haploidentical transplant donor was infused without further conditioning. Within 15 days of mobilized CD34-boost administration the patient showed complete resolution of both the aplasia and fungal infection. This case illustrates as proof-of-concept the efficacy and safety of selected CD34-stem cell boost from prior donor as salvage treatment of prolonged cytopenias after CAR-T cell therapy.


The use of two-dimensional Doppler sonography (color Doppler) in the diagnosis of hydranencephaly.
Hydranencephaly, first described by Cruveilhier, may be defined as a rare condition in which the cerebral hemispheres are replaced by membranous sacs filled with cerebrospinal fluid. Cerebral angiography is especially useful in differentiating hydranencephaly and severe hydrocephalus, massive subdural hygromas and alobar holoprosencephaly, but it is no better than other neuroradiological diagnosis methods in the neonate. This report outlines the diagnostic use of two-dimensional Doppler (color Doppler) in a child with hydranencephaly. Color Doppler was performed using an SSA-270A (Toshiba) with a 3.75 MHz phase-array sector scanning probe and no sedation. In this study, color Doppler demonstrated the internal carotid artery in the proximal portion, but the secondary and tertiary branches were not observed. In summary, color Doppler permitted early, easy diagnosis of hydranencephaly. In addition, the use of color Doppler was beneficial in diagnosing other anomalies of the central nervous system in neonates.


Persistent atrial fibrillation in heart failure with preserved ejection fraction: Prognostic relevance and association with clinical, imaging and invasive haemodynamic parameters.
Atrial fibrillation (AF) is a frequent finding in HFpEF. However, its association with invasive haemodynamics, imaging parameters and outcome in HFpEF is not well established. Furthermore, the relevance of AF subtype with regard to outcome is unclear. This study sought to investigate the prognostic impact of paroxysmal and persistent AF in a well-defined heart failure with preserved ejection fraction (HFpEF) population. Between 2010 and 2016, 254 HFpEF patients were prospectively enrolled. All patients underwent echocardiography as well as left and right heart catheterization. Patients without contraindications underwent CMR including T1 mapping. Follow-up and outcome data were collected. Patients with significant coronary artery disease were excluded. A total of 153 patients (60%) suffered from AF, 119 (47%) had persistent and 34 (13%) had paroxysmal AF. By multiple logistic regression analysis, persistent AF was independently associated with NT-proBNP (P = .003), NYHA functional class (P = .040), left and right atrial size (P = .022 and <.001, respectively), cardiac output (P = .002) and COPD (P = .034). After a median follow-up of 23 months (interquartile range 5-48), 92 patients (36%) reached the primary end point defined as hospitalization for heart failure or cardiovascular death. By multivariate Cox regression analysis, only persistent AF (P = .005) and six-minute walk distance (P = .011) were independently associated with the primary end point. Sixty percent of our HFpEF patients suffered from AF. Persistent but not paroxysmal AF was strongly associated with event-free survival and was independently related to NYHA functional class, serum NT-proBNP, atrial size, cardiac ouput and presence of COPD.


[Clinical usefulness of serum beta2-microglobulin determination in patients with multiple myeloma].
Serum concentrations of beta 2-microglobulin (beta 2M) were determined in 73 patients with various forms of multiple myeloma and in various phases of the proliferative process. These determinations showed that beta 2M may be a useful indicator of changes in tumour mass and proliferation activity, and also an important prognostic factor. In patients with active proliferation the serum beta 2M concentration was significantly higher than in the group with stable proliferative process, and particularly in remission. A correlation was found between the serum concentration of monoclonal protein and beta 2M concentration. In the group with the secretory form of myeloma significant differences were showed in the length of survival which depended on beta 2M concentration in serum. The median survival of patients with beta 2M concentration in serum below 5.0 mg/l was 52 months and that in those with this concentration above 8.0 mg/l was 24 months.


[Species-specific odor in closely related species: period of development in ontogenesis of house (Mus musculus musculus) and mound-building (Mus spicilegus) mice].
In two-choice odor tests, it was shown that house and mound-building mice began to discriminate the species-specific odor of young at the age of 17 days. The mice investigated the odor of conspecific urine significantly longer than that of heterospecific urine, when the young donors of urine, were 17 days old or older. Thus, the species-specific odor appears in ontogenesis of these species on the 17th day of life.


Foam counter-current chromatography with the cross-axis synchronous flow-through coil planet centrifuge.
A novel coil planet centrifuge system performs efficient foam separation in a long multilayer coil. Potential capability of the method was successfully demonstrated in separations of two sets of test samples using sodium dodecyl sulfate as a foaming agent. Rhodamine B with high foam affinity was quickly separated from Evans blue lacking foam affinity. A high concentration of salts such as disodium hydrogenphosphate in the surfactant solution effectively lowered the foam affinity of basic dyes, yielding complete peak resolution of rhodamine B and methylene blue according to a subtle difference in foam affinity.


Risperidone-associated hyperprolactinemia: evaluation in twenty psychiatric outpatients.
Despite the lack of extrapyramidal side effects, some atypical antipsychotics can induce an increase in prolactinemia, as the conventional neuroleptics do. We decided to evaluate the effects of risperidone on serum prolactin levels and prolactin-related adverse effects in 20 outpatients of an Italian community psychiatric service. Patients enrolled in this study were on risperidone (2-8 mg per day; mean dose=4.15+/-0.4 mg per day) treatment in the period May-November 2002. The 20 patients, 13 women and 7 men (mean age=36.38+/-3.2 years for women and 29.7+/-2.2 for men) who accomplished inclusion criteria, participated in the study after giving informed written consent. Raised prolactin levels were observed in 13 (9 women and 4 men) out of 20 patients, but only 8 patients presented prolactin-related adverse effects, libido reduction being the most frequent. In this observational study, risperidone enhanced serum prolactin in 65% of patients. A good correlation was found between age and prolactin levels in pre-menopausal women, although no clear correlation among duration of treatment, dose used, prolactin levels and prolactin-related adverse effects could be established.


Ventilation response to CO2 in birds. II. Contribution by intrapulmonary CO2 receptors.
The CO2 sensitivity of intrapulmonary CO2 receptors (IPC) in the duck was studied, before (Control) and after blockade of carbonic anhydrase by Diamox, by recording single unit afferent activity in the vagus nerve. During Control, IPC activity decreased with increasing airway CO2 concentration. After Diamox administration, the discharge from IPC was higher at all levels of airway PCO2, and the receptors' CO2 sensitivity was markedly attenuated. Comparing these results with measurements on ventilation and blood gases of the duck under similar experimental conditions (Powell et al., 1978b) suggests that IPC play a role in the adjustment of ventilation to altered concentrations of inspired CO2; IPC may thus be a significant component in the control of breathing under physiological conditions.


Adult ABO-incompatible liver transplantation by intraportal transfusion of donor-specific antigen: a case report.
A 55-year-old-woman suffering from fluminant hepatitis owing to autoimmune hepatitis underwent ABO-incompatible liver transplantation (LRLD) of blood type A to B. In this study, we investigated whether a new immunosuppressive strategy by intraportal transfusion of donor-specific leukocytes (DSLT) separated from whole blood would yield immunological benefit in adult ABO-LRLD. The operative course was uneventful; she was discharged at 46 days postoperatively without humoral or cellular rejection. On immunologic analysis, 54.6% intrahepatic macrochimerism of donor type CD56+ T cells was recognized at 1 month after transplantation. The interleukin-10 Th2 cytokine level was increased on postoperative day 1. Adult ABO-incompatible liver transplantation can be performed with acceptable patient and graft survival rates with a low risk of antibody-mediated rejection with our strategy of immunosuppression by intraportal administration of DSLT. Donor type CD56+ NKT cells may induce tolerance by a veto mechanism and/or an anti-idiotype network. ABO-incompatible liver transplantation may be improved by this strategy.


The effect of pirfenidone on rat chronic prostatitis/chronic pelvic pain syndrome and its mechanisms.
Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is an intractable problem of the urogenital system. The aetiopathogenesis and effective treatments for CP/CPPS are needed to be untangled. Pirfenidone is a molecule that exhibits anti-inflammatory, antifibrotic, and antioxidative stress capacities in a variety of animal experiments and clinical trials. This study was aimed to investigate the therapeutic effect of pirfenidone on CP/CPPS and to identify the mechanism responsible for it. A CP/CPPS model was induced in rats by intraprostatic injection of complete Freund's adjuvant (CFA). Blood and prostatic tissues were harvested for assessment after the administration of pirfenidone or vehicle for 4 weeks. The findings revealed that pirfenidone significantly ameliorated chronic pelvic pain and inhibited prostatic inflammation and fibrosis. Further study found that pirfenidone suppressed the expression of proinflammatory mediators, including tumor necrosis factor-α, interleukin-1β (IL-1β), IL-6, IL-8. Pirfenidone exhibited a potent antioxidant capacity through improving the activities of glutathione, catalase, total superoxide dismutase, and reducing the production of malondialdehyde. Furthermore, pirfenidone also facilitated the polarization of M2 macrophages and suppressed the activation of the nuclear factor-κB (NF-κB) signaling pathway. Pirfenidone can exert a beneficial effect against CFA-induced CP/CPPS by anti-inflammatory, antioxidative, antifibrotic properties, and the function is mediated at least partly through the M2 polarization of macrophages and the inhibition of NF-κB signaling pathway. These findings suggest that pirfenidone holds promise as a potential therapeutic for the treatment of CP/CPPS.


Development and validation of a physical activity monitor for use on a wheelchair.
Keeping physically active is important for people who mobilize using a wheelchair. However, current tools to measure physical activity in the wheelchair are either not validated or limited in their application. The purpose of this study was to develop and validate a monitoring system to measure wheelchair movement. The system developed consisted of a tri-axial accelerometer placed on the wheel of a wheelchair and an analysis algorithm to interpret the acceleration signals. The two accelerometer outputs in the plane of the wheel were used to calculate the angle of the wheel. From this, outcome measures of wheel revolutions, absolute angle and duration of movement were derived and the direction of movement (forwards or backwards) could be distinguished. Concurrent validity was assessed in comparison with video analysis in 14 people with spinal cord injury using their wheelchair on an indoor track and outdoor wheelchair skills course. Validity was assessed using intraclass correlation coefficients (ICC(2,1)) and Bland-Altman plots. The monitoring system demonstrated excellent validity for wheel revolutions, absolute angle and duration of movement (ICC(2,1)>0.999, 0.999, 0.981, respectively) in both manual and powered wheelchairs, when the wheelchair was propelled forwards and backwards, and for movements of various durations. This study has found this monitoring system to be an accurate and objective tool for measuring detailed information on wheelchair movement and maneuvering regardless of the propulsion technique, direction and speed.


Extracellular or intracellular application of argiotoxin-636 has inhibitory actions on membrane excitability and voltage-activated currents in cultured rat sensory neurones.
The whole cell variant of the patch clamp technique was used to investigate the actions of the polyamine amide spider toxin, argiotoxin-636, on the excitability of cultured dorsal root ganglion neurones. Synthesized argiotoxin-636 (0.1-100 microM) reduced neuronal excitability when applied to the extracellular environment by low pressure ejection or to the intracellular environment via the patch pipette solution. The toxin prolonged the duration of evoked action potentials and reduced the peak amplitude of action potentials. Intracellular and extracellular application of argiotoxin-636 also decreased the number of action potentials evoked in response to 800-ms depolarizing current commands. This action of the toxin was mimicked by 100 microM tetraethylammonium. Extracellular application of argiotoxin-636 inhibited voltage-activated K currents in a dose-dependent manner over the complete voltage range. This inhibition occurred without any significant changes in the voltage dependence of activation or inactivation. Intracellular application of argiotoxin-636, during 5-10 min of whole cell recording, also inhibited voltage-activated K+ currents without changing the voltage dependence of activation or steady-state inactivation. Extracellular or intracellular spermidine (250 microM) reversibly attenuated the inhibitory actions of extracellular argiotoxin-636. Argiotoxin-636 also inhibited voltage-activated Na + currents; this effect was dependent on repeated activation of the currents and the period during which the neurones were in culture. We conclude that application of argiotoxin-636 to either the extracellular or intracellular environment reduced excitability of cultured sensory neurones from neonatal rats and that this involved inhibition of both voltage-activated K+ and Na+ currents. The data suggest that the toxin was more effective at attenuating action potentials when neurones were repeatedly excited, and that access to inhibitory sites of action on the voltage-activated ion channels can be achieved from the inside of the neurone.


Proteomic profiling of postmortem prefrontal cortex tissue of suicide completers.
Suicide is a leading cause of death worldwide, presenting a serious public health problem. We aimed to investigate the biological basis of suicide completion using proteomics on postmortem brain tissue. Thirty-six postmortem brain samples (23 suicide completers and 13 controls) were collected. We evaluated the proteomic profile in the prefrontal cortex (Broadmann area 9, 10) using tandem mass tag-based quantification with liquid chromatography-tandem mass spectrometry. Bioinformatics tools were used to elucidate the biological mechanisms related to suicide. Subgroup analysis was conducted to identify common differentially expressed proteins among clinically different groups. Of 9801 proteins identified, 295 were differentially expressed between groups. Suicide completion samples were mostly enriched in the endocannabinoid and apoptotic pathways (CAPNS1, CSNK2B, PTP4A2). Among the differentially expressed proteins, GSTT1 was identified as a potential biomarker among suicide completers with psychiatric disorders. Our findings suggest that the previously under-recognized endocannabinoid system and apoptotic processes are highly involved in suicide.


Effects of follower load and rib cage on intervertebral disc pressure and sagittal plane curvature in static tests of cadaveric thoracic spines.
The clinical relevance of mechanical testing studies of cadaveric human thoracic spines could be enhanced by using follower preload techniques, by including the intact rib cage, and by measuring thoracic intervertebral disc pressures, but studies to date have not incorporated all of these components simultaneously. Thus, this study aimed to implement a follower preload in the thoracic spine with intact rib cage, and examine the effects of follower load, rib cage stiffening and rib cage removal on intervertebral disc pressures and sagittal plane curvatures in unconstrained static conditions. Intervertebral disc pressures increased linearly with follower load magnitude. The effect of the rib cage on disc pressures in static conditions remains unclear because testing order likely confounded the results. Disc pressures compared well with previous reports in vitro, and comparison with in vivo values suggests the use of a follower load of about 400N to approximate loading in upright standing. Follower load had no effect on sagittal plane spine curvature overall, suggesting successful application of the technique, although increased flexion in the upper spine and reduced flexion in the lower spine suggest that the follower load path was not optimized. Rib cage stiffening and removal both increased overall spine flexion slightly, although with differing effects at specific spinal locations. Overall, the approaches demonstrated here will support the use of follower preloads, intact rib cage, and disc pressure measurements to enhance the clinical relevance of future studies of the thoracic spine.


Photophysical studies of pheophorbide a and pheophytin a. Phosphorescence and photosensitized singlet oxygen luminescence.
The triplet states of pheophorbide a and pheophytin a were studied in several environments by direct measurement of the phosphorescence of the pigments and photosensitized singlet oxygen (1O2) luminescence. The spectra, lifetimes and quantum yields of phosphorescence and the quantum yields of 1O2 generation were determined. These parameters are similar for monomeric molecules of both pigments in all the environments studied. Aggregation of the pigment molecules leads to a strong decrease in the phosphorescence and 1O2 luminescence intensities, which is probably due to a large decrease in the triplet lifetime and triplet quantum yield in the aggregates. The results obtained for pheophorbide a and pheophytin a are compared with those previously reported for chlorophyll alpha. The data suggest that the photodynamic activity of the pigments in living tissues is probably determined by the monomeric pigment molecules formed in hydrophobic cellular structures. Aggregated molecules seem to have a much lower activity.


Increased RUNX3 expression mediates tumor-promoting ability of human breast cancer-associated fibroblasts.
Cancer-associated fibroblasts (CAFs) are a major stromal component of human breast cancers and often promote tumor proliferation, progression and malignancy. We previously established an experimental CAF (exp-CAF) cell line equipped with a potent tumor-promoting ability. It was generated through prolonged incubation of immortalized human mammary fibroblasts with human breast cancer cells in a tumor xenograft mouse model. Herein, we found that the exp-CAFs highly express Runt-related transcription factor 3 (RUNX3), while counterpart fibroblasts do not. In breast cancer patients, the proportion of RUNX3-positive stromal fibroblast-like cells tends to be higher in cancerous regions than in non-cancerous regions. These findings suggest an association of RUNX3 with CAF characteristics in human breast cancers. To investigate the functional role of RUNX3 in CAFs, the exp-CAFs with or without shRNA-directed knockdown of RUNX3 were implanted with breast cancer cells subcutaneously in immunodeficient mice. Comparison of the resulting xenograft tumors revealed that tumor growth was significantly attenuated when RUNX3 expression was suppressed in the fibroblasts. Consistently, Ki-67 and CD31 immunohistochemical staining of the tumor sections indicated reduction of cancer cell proliferation and microvessel formation in the tumors formed with the RUNX3-suppressed exp-CAFs. These results suggest that increased RUNX3 expression could contribute to the tumor-promoting ability of CAFs through mediating cancer cell growth and neoangiogenesis in human breast tumors.


Role of CRISPR/Cas9 in the treatment of Duchenne muscular dystrophy and its delivery strategies.
Duchenne muscular dystrophy (DMD) is a neuromuscular disorder brought on by mutations in the DMD gene, which prevent muscle cells from expressing the dystrophin protein. CRISPR/Cas9 technology has evolved as potential option to treat DMD due to its ability to permanently skip exons, restoring the disrupted DMD reading frame and leading to dystrophin restoration. Even though, having potential to treat DMD, the delivery, safety and efficacy of this technology is still challenging. Several delivery methods, including viral vectors, nanoparticles, and electroporation, have been explored to deliver CRISPR/Cas9 to the targeted cells. Despite the potential of CRISPR/Cas9 technology in the treatment of DMD, several limitations need to be addressed. The off-target effects of CRISPR/Cas9 are a major concern that needs to be addressed to avoid unintended mutations. The delivery of CRISPR/Cas9 to the target cells and the immune response due to the viral vectors used for delivery are a few other limitations. The clinical trials of CRISPR/Cas9 for DMD provide valuable insights into the safety and efficacy of this technology in humans and the limitations that need to be known. Therefore, in this review we insightfully discussed the challenges and limitations of CRISPR/Cas9 in the treatment of DMD and delivery strategies used, and the ongoing efforts to overcome these challenges and restore dystrophin expression in DMD patients in the ongoing trials.


Enhanced photocatalytic hydrogen evolution by piezoelectric effects based on MoSe2/Se-decorated CdS nanowire edge-on heterostructure.
The build-in electric field by the construction of heterojunction is one of the most promising strategies to suppress the recombination of photogenerated carriers. Here, we reported a piezo-photocatalytic system composed of Se-decorated CdS nanowires and few-layered edge-on MoSe2 nanosheets for efficient H2 generation by two-pot hydrothermal synthesis. The few-layered MoSe2 exposed abundant edge sites for hydrogen evolution reaction (HER). The activity of 20-MoSe2/CdS0.95Se0.05 (20-MS/CSS, with 20 mol% of MoSe2 loading) nanocomposite casted a remarkable photocatalytic HER performance, with a rate of 47.3 mmol h-1 g-1. Moreover, MoSe2 nanosheets deformed to generate the piezoelectric polarization field under magnetic stirring, which rendered efficient separation of photogenerated carriers, resulting in a piezo-photocatalytic synergistic effect. As a result, the HER of 20-MS/CSS at 900 rpm for piezo-photocatalysis was 59.1 mmol h-1 g-1, which was 1.25 times that of 20-MS/CSS for photocatalysis. Meanwhile, the photoelectrochemical measurements further visualized the piezo-photoelectric synergy. This study exposes a new way for utilizing mechanical energy to improve photocatalytic performance, and achieving high piezo-photocatalysis.


Out of destruction comes new growth: Pore-forming antimicrobials make pancreas grow.
Gut-residing bacteria are known to regulate the physiologies of distal organs. However, the mechanism behind the long-distance communication between gut microbes and distal organs remains unknown. In this issue of Cell Metabolism, two studies show that β cell expansion in the pancreas depends on bacterially induced antimicrobials produced in the gut.


Individual stress and its relationship to termination of nurse training.
During interviews with 23 learners (student and pupil nurses) who left training as a response to stress, the following points were explored; knowledge of nursing before commencing training; reasons for wanting to nurse; situations which produced stress; the way in which individual stress responses were expressed; and methods for coping with stress. Clinical experiences had produced the most distress, related predominantly to negative attitudes of other nurses. Behavioural and physiological signs of stress were observed as well as psychological reactions such as worry. People had few adaptive coping strategies and received little support in their time of need. The majority expressed a desire to try nursing again.


Multifactorial control of reproductive and growth axis in male goldfish: Influences of GnRH, GnIH and thyroid hormone.
Reproduction and growth are under multifactorial control of neurohormones and peripheral hormones. This study investigated seasonally related effects of GnIH, GnRH, and T3 on the reproductive and growth axis in male goldfish at three stages of gonadal recrudescence. The effects of injection treatments with GnRH, GnIH and/or T3 were examined by measuring serum LH and GH levels, as well as peripheral transcript levels, using a factorial design. As expected, GnRH elevated serum LH and GH levels in a seasonally dependant manner, with maximal elevations of LH in late stages of gonadal recrudescence (Spring) and maximal increases in GH in the regressed gonadal stage (Summer). GnIH injection increased serum LH and GH levels only in fish at the regressed stage but exerted both stimulatory and inhibitory effects on GnRH-induced LH responses depending on season. T3 treatment mainly had stimulatory effects on circulating LH levels and inhibitory effects on serum GH concentrations. In the liver and testes, we observed seasonal differences in thyroid receptors, estrogen receptors, vitellogenin, follicle-stimulating hormone receptor, aromatase and IGF-I transcript levels that were tissue- and sex-specific. Generally, there were no clear correlation between circulating LH and GH levels and peripheral transcript levels, presumably due to time-related response and possible direct interaction of GnRH and GnIH at the level of liver and testis. The results support the hypothesis that GnRH and GnIH are important components of multifactorial mechanisms that work in concert with T3 to regulate reciprocal control of reproduction and growth in goldfish.


Reversible truncal vagotomy in conscious dogs.
Prior attempts at in vivo reversible vagal denervation of the gastrointestinal tract have been limited to cervical cooling techniques that also denervate both sympathetic and vagal pulmonary and cardiac branches. Denervation of vagal efferent fibers at this level has produced results that are inconsistent with those obtained after surgical truncal vagotomy. We have, therefore, developed a technique to provide reversible vagal denervation below the pulmonary and cardiac branches for the study of gastric motility. Five dogs, previously equipped with gastric strain gages and electrodes, underwent implantation of a tubular cooling jacket around a distal thoracic vagal trunk with contralateral vagotomy (4 dogs), or around both vagal trunks (1 dog). The jacket was made of stainless steel tubing in a "J" design. Its inside channel was lined with a sterling silver sheet, and a thermistor was attached to record temperature change. Silicone tubing coursed externally to a pump and flask to which 95% ethanol at -70 degrees C was circulated at variable speeds. Thoracic vagal cooling, extending up to 5 h, reversibly blocked gastric contractions induced by insulin hypoglycemia. Contractile waves were terminated at device temperatures of 2 degrees-6 degrees C but promptly returned with warming. Dogs were tranquil during denervation, and enclosed nerves remained functional for greater than 40 days.


Antioxidant properties of new chalcogenides against lipid peroxidation in rat brain.
Ebselen (2-phenyl- 1,2-benzisoselenazole-3 (2H)-one) is a seleno-organic compound with antioxidant properties, and anti-inflammatory actions. Recently, ebselen improved the outcome of acute ischemic stroke in humans. In the present study, the potential antioxidant capacity of organochalcogenide compounds diphenyl diselenide (PhSe)2, diphenyl ditelluride (PhTe)2, diphenyl disulfide (PhS)2, p-Cl-diphenyl diselenide (pCl-PhSe)2, bis-[S-4-isopropyl 2-phenyl oxazoline] diselenide (AA-Se)2, bis-[S-4-isopropyl 2-phenyl oxazoline] ditelluride (AA-Te)2 and bis-[S-4-isopropyl 2-phenyl oxazoline] disulfide (AA-S)2 was compared with that of ebselen (a classical antioxidant). Spontaneous and quinolinic acid (QA)- (2 mM) and sodium nitroprusside (SNP)- (5 microM)-induced thiobarbituric reactive species (TBARS) production by rat brain homogenates was determined colorimetrically. TBARS formation was reduced by ebselen, (PhSe)2, (PhTe)2, (AA-Se)2, (AA-S)2 and (pCl- PhSe)2 to basal rates. The concentrations of these compounds needed to inhibit TBARS formation by 50% (IC50) are 1.71 microM, 3.73 microM, 1.63 microM, 9.85 microM, >33.3 microM, 23.2 microM and 4.83 microM, respectively for QA. For TBARS production induced by SNP the IC50 was 2.02 microM, 12.5 microM, 2.80 microM, >33.3 microM, 24.5 microM and 7.55 microM, respectively. The compounds (AA-Te)2 and (PhS)2 have no antioxidant activity and pro-oxidant activity, respectively. These results suggest that (AA-Se)2 and (AA-S)2 can be considered as potential pharmaceutical antioxidant agents.


Synthetic physical contact-remodeled rhizosphere microbiome for enhanced phytoremediation.
Phytoremediation is a prevalent strategy to treat environmental pollution caused by heavy metals and eutrophication-related pollutants. Although rhizosphere microbiome is critical for phytoremediation, it remains a great challenge to artificially remodel rhizosphere microbiome for enhancing multiple pollutant treatment. In this study, we designed a synthetic bacterium to strengthen physical contact between natural microbes and plant roots for remodeling the Eichhornia crassipes rhizosphere microbiome during phytoremediation. The synthetic bacterium EcCMC was constructed by introducing a surface-displayed synthetic protein CMC composed of two glucan-binding domains separated by the sequence of the fluorescent protein mCherry. This synthetic bacterium strongly bound glucans and recruited natural glucan-producing bacterial and fungal cells. Microbiome and metabolomic analysis revealed that EcCMC remarkably remodeled rhizosphere microbiome and increased stress response-related metabolites, leading to the increased activity of antioxidant enzymes involved in stress resistance. The remodeled microbiome further promoted plant growth, and enhanced accumulation of multiple pollutants into the plants, with the removal efficiency of the heavy metal cadmium, total organic matters, total nitrogen, total potassium, and total phosphorus reaching up to 98%, 80%, 97%, 93%, and 90%, respectively. This study sheds a novel light on remodeling of rhizosphere microbiome for enhanced phytoremediation of water and soil systems.


Mechanisms of tumor necrosis factor-alpha and interleukin-6 induction during human liver transplantation.
In human orthotopic liver transplantation (LTX) intraoperative elevations of TNF-alpha (> 100 pg/ml) and IL-6 (>800 pg/ml) have been found to correlate with early post-operative rejections and infections respectively. In this study the possible mechanism responsible for the induction of these cytokines has been investigated during liver allografting in 38 recipients. Intraoperative elevations of TNF-alpha (> 100 pg/ml) were detected in the majority of pre-transplant endotoxin positive recipients (8/12, > 10 endotoxin units/ml), the patients turning endotoxin positive until the end of grafting (3/5), and in a subgroup (6/21 patients), apparently endotoxin negative for the whole operation. Therefore endotoxin (ET) seems to stimulate release of TNF-alpha in approximately 50% of the patients, whereas sensitized Kupffer graft cells or immediate allograft reactivity of the host are likely to account for the remaining TNF-alpha positive cases. Elevations of IL-6 > 800 pg/ml) were found in approximately 50% of the TNF-alpha positive cases, indicating partially independent regulatory pathways for IL-6 induction in the TNF-alpha negative patients. In agreement with a previous study, 11/13 (85%) of the intraoperative TNF-alpha positive recipients rejected their grafts within the first 10 days post-operatively. These data demonstrate that ET/infection associated as well as ET independent/reperfusion associated intraoperative TNF-alpha elevations, promote the initiation of allograft rejection in human liver transplantation. The transient and low endotoxaemia caused by the liver grafting procedure performed without veno-venous bypass seems to be of minor importance in the intraoperative induction of TNF-alpha.


A study of PVRL1 mutations for non-syndromic cleft lip and/or palate among Taiwanese patients.
Mutations of codons 185 and 323, especially the W185X mutation, of the PVRL1 gene among non-syndromic cleft lip and/or palate (CL/P) patients and normal controls in Taiwan were studied in order to determine whether there are mutations that play a part in the formation of non-syndromic CL/P. A total of 76 patients were enrolled; 66 sporadic non-syndromic CL/P patients and 10 normal controls. The mutation survey for codons 185 and 323 was conducted using a polymerase chain reaction and DNA sequencing. Neither mutations of codons 185 and 323 were noted for any of the 76 patients (152 alleles), nor were found any other mutations in either exon 3 or 5 of the PVRL1 gene. These results suggest that mutations of the PVRL1 gene at codons 185 and 323, especially the W185X mutation, do not participate in the formation of CL/P within the Taiwanese population examined.


Evaluation of several dosing regimens of cefepime, with various simulations of renal function, against clinical isolates of Pseudomonas aeruginosa in a pharmacodynamic infection model.
The objectives of this study were as follows: (i) to examine the killing activity of 2-g doses of cefepime against two clinical isolates (mucoid and nonmucoid) of Pseudomonas aeruginosa in a pharmacodynamic in vitro infection model, (ii) to compare the percentage of time above the MIC (T > MIC) for each of the regimens against P. aeruginosa, and (iii) to evaluate the area under the bactericidal curve for each regimen. Cefepime was administered at intervals of 8, 12, and 24 h with and without tobramycin, and two different levels of renal function were simulated: normal (creatinine clearance [CLCR] = 90 ml/min) and decreased (CRCL = 60 ml/min). Also, the killing activity of cefepime with and without tobramycin was compared to the killing activity of ceftazidime (2 g every 8 h) with and without tobramycin. The T > MIC was 100% in the central chamber except for the regimen in which cefepime was administered every 12 h and the CLCR was 90 ml/min, which provided concentrations above the MIC for 92% of the dosing interval against the C31 (mucoid; MIC of cefepime, 4 microg/ml) isolate and for 75% of the interval against the C34 (nonmucoid; MIC of cefepime, 8 microg/ml) isolate. All cefepime and ceftazidime monotherapy simulations resulted in 99.9% killing of the nonmucoid isolate within 4 to 8 h and within 4 to 6 h, respectively. Against the mucoid isolate, 99.9% killing was achieved only with combination therapy. The results of this study indicate that cefepime dosed at 2 g every 12 h under conditions of normal renal function and every 24 h with decreased creatinine clearance (60 ml/min) is effective both as monotherapy and in combination therapy against a nonmucoid strain of P. aeruginosa. With cefepime MICs of 4 and 8 microg/ml, the single-agent regimens provided T > MIC values in the central chamber for 92 and >/=75% of the dosing interval against the mucoid and nonmucoid isolates, respectively. Cefepime dosed at 2 g every 12 h, with a creatinine clearance of 90 ml/min, and every 24 h, with a creatinine clearance of 60 ml/min, resulted in killing activity equivalent to that of ceftazidime dosed at 2 g every 8 h. None of the monotherapies provided adequate killing of the mucoid strain of P. aeruginosa despite drug concentrations being above the MIC for >/=92% of all dosing intervals. Finally, combination therapy with tobramycin and either cefepime or ceftazidime enhanced the killing of both the mucoid and nonmucoid P. aeruginosa isolates.


Epigenetic modifications in diabetes.
Diabetes is now considered as a 'silent epidemic' that claims over four million lives every year, and the disease knows no socioeconomic boundaries. Despite extensive efforts by the National and International organizations, and cutting-edge research, about 11% world's population is expected to suffer from diabetes (and its complications) by year 2045. This life-long disease damages both the microvasculature and the macrovasculature of the body, and affects many metabolic and molecular pathways, altering the expression of many genes. Recent research has shown that external factors, such as environmental factors, lifestyle and pollutants can also regulate gene expression, and contribute in the disease development and progression. Many epigenetic modifications are implicated in the development of micro- and macro- vascular complications including DNA methylation and histone modifications of several genes implicated in their development. Furthermore, several noncoding RNAs, such as micro RNAs and long noncoding RNAs, are also altered, affecting many biochemical pathways. Epigenetic modifications, however, have the advantage that they could be passed to the next generation, or can be erased. They are now being explored as therapeutical target(s) in the cancer field, which opens up the possibility to use them for treating diabetes and preventing/slowing down its complications.


Neonatal exposure to technical methoxychlor alters pregnancy outcome in female mice.
This study was designed to determine the ability of female mice who were exposed neonatally to the pesticide methoxychlor (MXC) to mate, ovulate, and become pregnant upon reaching sexual maturity. One-day-old female mice (5 to 8/group) were exposed daily by intraperitoneal (ip) injection for 14 d to either sesame oil or 10 microg estradiol-17beta or 0.1, 0.5 or 1.0 mg MXC suspended in sesame oil. The MXC exposures corresponded to 14 to 71, 68 to 357, or 135 to 714 mg/kg body weight, respectively. Three months later, female mice were placed with proven breeder males and checked daily for vaginal plugs. Mated female mice were sacrificed 18 d after the appearance of a vaginal plug to evaluate pregnancy. Uteri were examined for the presence of living fetuses and/or resorption sites. Ovaries were removed and prepared for histologic evaluation and tabulation of corpora lutea. All mice from all three MXC-treated groups did in fact mate, in comparison with only one of those exposed neonatally to estradiol. Increasing the dose of MXC produced a decreased number of pregnant animals at 18 d following mating. The mean number of live fetuses/litter was reduced in the 0.5 and 1.0 mg MXC-treated groups. Corpora lutea were significantly reduced in ovaries from only the 1.0 mg MXC group and the estradiol group. No effects of treatment were seen at 0.1 mg MXC. It is concluded that neonatal exposure to MXC does not interfere with mating. Instead, significant alterations are seen in initiating and/or maintaining pregnancy. The deleterious effects on pregnancy may be due to the influence of neonatal MXC treatments on the hypothalamic-pituitary-ovarian axis as well as on possible alteration of the uterine environment.


Lipin1 is a key factor for the maturation and maintenance of adipocytes in the regulatory network with CCAAT/enhancer-binding protein alpha and peroxisome proliferator-activated receptor gamma 2.
Lipin1 expression was induced at a late stage of differentiation of 3T3-L1 preadipocytes and maintained at high levels in mature adipocytes. Knockdown of expression of lipin1 by small interfering RNA in 3T3-L1 preadipocytes almost completely inhibited differentiation into adipocytes, whereas overexpression of lipin1 accelerated adipocyte differentiation, demonstrating that lipin1 is required for adipocyte differentiation. In mature adipocytes, transfection of lipin1-small interfering RNA decreased the expression of adipocyte functional genes, indicating the involvement of lipin1 in the maintenance of adipocyte function. Lipin1 increases the transcription-activating function of peroxisome proliferator-activated receptor gamma(2) (PPAR gamma(2)) via direct physical interaction, whereas lipin1 did not affect the function of other adipocyte-related transcription factors such as C/EBP alpha, liver X-activated receptor alpha, or sterol regulatory element binding protein 1c. In mature adipocytes, lipin1 was specifically recruited to the PPAR gamma-response elements of the phosphoenolpyruvate carboxykinase gene, an adipocyte-specific gene. C/EBP alpha up-regulates lipin1 transcription by directly binding to the lipin1 promoter. Based on the existence of a positive feedback loop between C/EBP alpha and PPAR gamma(2), we propose that lipin1 functions as an amplifier of the network between these factors, resulting in the maintenance of high levels of the specific gene expression that are required for adipogenesis and mature adipocyte functions.


Do surgical margins matter after mastectomy? A systematic review.
No consensus exists regarding adequacy of margins after mastectomy. To determine if pathological margin proximity is associated with local (LR) or distant recurrence after mastectomy for early invasive breast cancer or ductal carcinoma in situ. A systematic review of literature published from 1980 to 2019 and meta-analysis was conducted. Unpublished data were sought from authors (PROSPERO (CRD42019127541)). Thirty-four studies comprising 34,833 breast cancer patients were included in the quantitative synthesis. Eligible studies reported on patients undergoing curative mastectomy for cancer allowing estimation of outcomes in relation to margin status/width. The association between pathological margin status and local (LR) and distant recurrence was considered using random effects modelling. PRISMA guidelines were followed. Positive margins were associated with increased LR on multivariable analyses (HR, 2·64, (95%CI 2·01-3·46)) and LR was higher regardless of the distance of tumour from the margin defined as positive. After skin-sparing mastectomy, positive margins were associated with increased LR (HR 3·40, (95%CI 1·9-6·2)). In the 4 studies reporting distant recurrence, patients with involved margins had a higher risk (HR 1·53, (95%CI 1·03-2·25)). Failure to achieve clear margins after mastectomy may increase the risks of local and distant recurrence. Adequate margin clearance should be recommended to minimize recurrence after mastectomy in National and International Guidelines.


Faster engraftment of neutrophils and platelets with peripheral blood stem cells from unrelated donors: a comparison with marrow transplantation.
Engraftment was achieved in 43/45 (95%) recipients of peripheral blood stem cells (PBSC) from HLA-compatible unrelated donors (n = 45), compared to all 45 patients in matched controls receiving bone marrow and 14/18 (78%) recipients of CD34-selected PBSC (P < 0.01). The time to reach ANC >0.5 x 10(9)/l was a median of 16 days in the PBSC and CD34 groups, compared to 20 days in the bone marrow controls (P < 0.001 vs PBSC). The time to reach platelets >50 x 10(9)/l was a median of 23 days in the PBSC group and 24 days in the CD34 group, which was significantly faster than 29 days in the bone marrow controls (P < 0.01). Acute GVHD grades II-IV developed in 30% in the PBSC group, 20% in the recipients of bone marrow and 18% in the CD34 group. The corresponding figures for chronic GVHD were 59%, 85% and 0% (P < 0.01) in the three groups, respectively. The probability of non-relapse death was 27% in the recipients of PBSC, 21% in the bone marrow controls and 60% in the CD34 group (NS). The 2-year leukaemia-free survival was 46% in the PBSC group, 41% in the bone marrow group and 25% in the CD34 group (NS).


Multicenter Validation of the CamGFR Model for Estimated Glomerular Filtration Rate.
Important oncological management decisions rely on kidney function assessed by serum creatinine-based estimated glomerular filtration rate (eGFR). However, no large-scale multicenter comparisons of methods to determine eGFR in patients with cancer are available. To compare the performance of formulas for eGFR based on routine clinical parameters and serum creatinine not calibrated with isotope dilution mass spectrometry, we studied 3620 patients with cancer and 166 without cancer who had their glomerular filtration rate (GFR) measured with an exogenous nuclear tracer at one of seven clinical centers. The mean measured GFR was 86 mL/min. Accuracy of all models was center dependent, reflecting intercenter variability of isotope dilution mass spectrometry-creatinine measurements. CamGFR was the most accurate model for eGFR (root-mean-squared error 17.3 mL/min) followed by the Chronic Kidney Disease Epidemiology Collaboration model (root-mean-squared error 18.2 mL/min).


Fos-immunoreactivity in the hypothalamus: dependency on the diurnal rhythm, sleep, gender, and estrogen.
Diurnal variations and sleep deprivation-induced changes in the number of Fos-immunoreactive (Fos-IR) neurons in various hypothalamic/preoptic nuclei were studied in the rat. The nuclei implicated in sleep regulation, the ventrolateral preoptic (VLPO), median preoptic (MnPO), and suprachiasmatic (SCN, dorsomedial subdivision) nuclei, displayed maximum c-fos expression in the rest (light) period. Sleep deprivation (S.D.) suppressed Fos-IR in the dorsomedial subdivision of SCN but failed to alter Fos in the VLPO. Fos-IR increased in the VLPO during recovery after S.D. A nocturnal rise in Fos expression was detected in the arcuate (ARC), anterodorsal preoptic (ADP) and anteroventral periventricular (AVPV) nuclei whereas the lateroanterior hypothalamic nucleus (LA) and the ventrolateral subdivision of SCN did not display diurnal variations. S.D. stimulated Fos expression in the ARC, ADP, and LA. Statistically significant, albeit modest, differences were noted in the number of Fos-IR cells between males and cycling female (estrus/diestrus) in the VLPO, MnPO, ARC, LA, and AVPV, and the female ADP did not display diurnal variations. Ovariectomy (OVX) was followed by marked reduction in Fos expression in the VLPO, SCN, and AVPV, and the diurnal rhythm decreased in the VLPO, and vanished in the dorsomedial SCN, and AVP. Estrogen administration to OVX female rats stimulated Fos expression in most nuclei, and the lost diurnal variations reoccurred. In contrast, castration of male rats had little effect on Fos expression (slight rises in diurnal Fos in the ARC and ventrolateral SCN). The results suggest that Fos expression is highly estrogen-dependent in many hypothalamic nuclei including those that have been implicated in sleep regulation.


Effect of diamide on isolated guinea pig atria.
Diamide [(diazinedicarboxylic acid-bis (dimethylamide)] 50 micrograms/ml induced a positive inotropic and chronotropic effect on spontaneously beating atria from normal and reserpine pretreated guinea pigs and a positive inotropic action on electrically driven left atria. Cumulative addition of diamide 600 micrograms/ml caused a systolic standstill of atria. D(-) INPEA 2 micrograms/ml, a beta-adrenergic blocking agent, antagonized the positive chronotropic action of diamide 50 micrograms/ml while it did not affect the positive inotropic action caused by diamide. In isolated atria perfused with Ca2+ free medium and recovered by stepwise addition of calcium chloride, the calcium level required to restore normal activity was 1.04 mmol/1 while it was 0.52 mmol/l in the presence of diamide. Ruthenium red 10 micrograms/ml, a specific inhibitor of calcium transport in mitochondria, antagonized diamide-induced recovery of the cardiac activity in calcium poor medium. Two different mechanisms, that involve calcium movements and beta-receptor, have been proposed for diamide action on heart muscle.


Epileptic syndromes in infancy and childhood.
The aim of this article is to review new epilepsy syndromes, acquire a new understanding of older ones and emphasize the impact of this concept on basic research regarding aetiology and treatment. In addition to those included in the classification of the International League Against Epilepsy, new epilepsy syndromes comprise febrile seizures plus, benign familial neonatal-infantile seizures (BFNIS), benign infantile focal epilepsy with midline spikes and waves during sleep (BFIS), malignant migrating partial seizures in infancy, devastating epilepsy in school age children and late onset cryptogenic spasms. Genetics played a central role in identifying some new entities (BFNIS, BFIS with choreoathetosis), to delineate older syndromes (Dravet syndrome and myoclonic astatic epilepsy) and determine their mechanisms (infantile spasms, pyridoxine dependent seizures, neonatal encephalopathy with suppression bursts). A significant number of children, mainly infants, do not fit in any of the described epilepsy syndromes. Still many patients with infantile epilepsy require the identification of cause or recognition of an epilepsy syndrome.


The summer I learned to write.
A summer course in English grammar challenged a registered nurse student to learn new behaviors and acquire effective writing skills. A confident nursing writer now, the author shares her experience for registered nurse students who continue to face some of the same challenges.


Below-knee amputation using the sagittal technique: a comparison with the coronal amputation.
Fourteen patients had below-knee amputation performed by the coronal technique and 17 by the sagittal technique. By changing the incision to a sagittal plane, a substantial improvement is observed in wound morbidity and in earlier time of definitive prosthetic wear.


Biomolecular condensates - extant relics or evolving microcompartments?
Unprecedented discoveries during the past decade have unearthed the ubiquitous presence of biomolecular condensates (BCs) in diverse organisms and their involvement in a plethora of biological functions. A predominant number of BCs involve coacervation of RNA and proteins that demix from homogenous solutions by a process of phase separation well described by liquid-liquid phase separation (LLPS), which results in a phase with higher concentration and density from the bulk solution. BCs provide a simple and effective means to achieve reversible spatiotemporal control of cellular processes and adaptation to environmental stimuli in an energy-independent manner. The journey into the past of this phenomenon provides clues to the evolutionary origins of life itself. Here I assemble some current and historic discoveries on LLPS to contemplate whether BCs are extant biological hubs or evolving microcompartments. I conclude that BCs in biology could be extant as a phenomenon but are co-evolving as functionally and compositionally complex microcompartments in cells alongside the membrane-bound organelles.


Prevalence and demographic characteristics of vulvodynia in a population-based sample.
The objective of the study was to determine the prevalence and characteristics of vulvodynia among women in southeast Michigan. A population-based study of adult women was conducted, using telephone recruitment and completion of a self-administered survey. Weighted estimates of vulvodynia prevalence and characteristics were determined. Over a year, 2542 women were recruited and 2269 (89.3%) completed the self-administered survey. The weighted prevalence of vulvodynia was 8.3% (95% confidence interval, 7.0-9.8) or approximately 101,000 women in the targeted population. Prevalence remained stable through age 70 years and declined thereafter. Among sexually active women, the prevalence was similar at all ages. Of 208 women meeting vulvodynia criteria, 101 (48.6%) had sought treatment, and only 3 (1.4%) had been diagnosed with vulvodynia (unweighted values). Previous vulvodynia symptoms had resolved in 384 women (16.9%) after a mean duration of 12.5 years. Vulvodynia is common, although rarely diagnosed. Prevalence remains high among sexually active women of any age.


Hand grip strength and early mortality after hip fracture.
This research describes the risk of death in elderly after hip fracture according to their strength, measured by hand grip. The result is that the weaker the patient, the greater the risk of death after hip fracture, highlighting the need to assess the force in those patients. For the coming years, most of hip fractures will occur in developing countries. It has been described that low muscular strength, measured by grip strength, increases the risk of mortality in those with hip fracture, in both high-and low- income countries. The objective of this study was to determine the mortality among patients with hip fracture and lower hand grip strength (HGS). We conducted a cohort and longitudinal study at Hip and Pelvic Surgery Department of a tertiary hospital, in Monterrey, Mexico. The study included patients aged over of 69, admitted for hip fracture surgery from February 1st 2013 to July 31st 2014. HGS measurement was performed by a trained physician at arrival to emergency department prior to surgery; clinimetric variables were asked, and a complete medical history was included. A total of 670 patients were included in the study and grouped in different tertiles according to hand grip strength. During follow-up, there were 112 deaths (17.4%), 61 (27.5%) in tertile 1, 37 (17.1%) in tertile 2, and 14 (6.8%) in tertile 3, p < 0.001. The association remained significant after adjusting for confounding variables. Less than 5% of patients discharged from hospital were identified with osteoporosis. Lower hand grip strength in patients with a hip fracture is associated with high mortality after hip fracture.


Understanding the patterns and mechanisms of urban water ecosystem degradation: phytoplankton community structure and water quality in the Qinhuai River, Nanjing City, China.
The temporal and spatial distribution characteristics of environmental parameters and the phytoplankton community were investigated in October 2010 and January 2011 in the Qinhuai River, Nanjing, China. Results showed that the water quality in the study area was generally poor, and the main parameters exceeding standards (level V) were nitrogen and phosphorus. The observed average concentrations of the total nitrogen (TN) were 4.90 mg L(-1) in autumn and 9.29 mg L(-1) in winter, and those of the total phosphorus (TP) were 0.24 mg L(-1) in autumn and 0.88 mg L(-1) in winter, respectively. Thirty-seven species, 30 genera, and four phyla of phytoplankton were detected in the river. Cyanophyta and Bacillariophyta were the dominant phyla in autumn, with average abundance and biomass of 221.5 × 10(4) cells L(-1) and 4.41 mg L(-1), respectively. The dominant population in winter was Bacillariophyta, and the average abundance and biomass were 153.4 × 10(4) cells L(-1) and 6.58 mg L(-1), respectively. The results of canonical correspondence analysis (CCA) between environmental parameters and phytoplankton communities showed that Chlorophyta could tolerate the higher concentrations of the permanganate index, nitrogen, and phosphorus in eutrophic water; Bacillariophyta could adapt well to changing water environments; and the TN/TP ratio had obvious impacts on the distributions of Cyanophyta, Euglenophyta, and some species of Chlorophyta. CCA analyses for autumn and winter data revealed that the main environmental parameters influencing phytoplankton distribution were water temperature, conductivity, and total nitrogen, and the secondary factors were dissolved oxygen, NH4(+)-N, NO3-N, TN, CODMn, TN/TP ratio, and oxidation-reduction potential.


[Conditioned reactions elaborated on the basis of reinforcement by hypothalamic stimulation].
The possibility of elaboration of conditioned emotional reactions on the basis of the stimulation of the hypothalamus, was studied in cats. Reinforcement of a sound (500 c/s) by electrical stimulation of the hypothalamus, which produced reactions of undirected rage, of fear with avoidance, of food search, led to the formation of a conditioned passive defensive reaction, analogous to that based on pain reinforcement. Elaboration of conditioned emotional reactions were blocked by switching off of the sensorimotor cortex by cold. A conclusion is made that during stimulation of the hypothalamus, the state of fear serves as the reinforcing factor and that the sensorimotor neocortical areas participate in the estimation of biological significance of the conditioned signal.


Overweight children find food more reinforcing and consume more energy than do nonoverweight children.
The reinforcing value of food is a reliable index of motivation to eat and energy intake. Obese adults find food more reinforcing than do nonobese adults. The present study was designed to assess whether the relative reinforcing value of food differs as a function of weight status in 8-12-y-old children and whether the relative reinforcing value of food differs depending on the types of available nonfood alternatives. The reinforcing value of pizza (experiment 1) or snack foods (experiment 2) was measured on progressive ratio schedules of reinforcement in nonoverweight and overweight children. Experiment 2 also compared the relative reinforcing value of food and 2 nonfood alternatives: time to spend playing a hand-held video game or time to spend reading magazines or completing word searches or mazes. In both experiments, overweight children found food more reinforcing and consumed more energy than did their leaner peers. In experiment 2, the relative reinforcing value of food versus sedentary activity was higher in overweight children, but lower in nonoverweight children, regardless of the type of alternative activity available. These results show that overweight children find food more reinforcing than do nonoverweight children. This individual difference was replicated in different experiments using different types of foods and food alternatives. These studies provide support for studying food reinforcement as a factor associated with overweight and obesity.


Correction: Hokynar, K. et al. Chlamydia-Like Organisms (CLOs) in Finnish Ixodes ricinus Ticks and Human Skin. Microorganisms 2016, 4, 28.
The authors wish to make the following modification to this paper [...].


Qualitative detection of placidyl (ethchlorvynol) alone or in combination with poly-drugs in drug abuse urine screening programs using ion-exchange paper and/or liquid-liquid extraction.
Two procedures for the detection of ethchlorvynol are presented. Procedure I involves the use of SA-2-cation-exchange resin loaded paper. The use of multiple ion-exchange resin papers is proposed in a sequence that enables the detection of amphetamines, barbiturates, phenothiazines, propoxyphene (Darvon), phencyclidine, cocaine (benzoyl ecgonine), pentazocine (Talwin) and benzodiazepines in addition to ethchlorvynol. This procedure discusses the details of detecting ethchlorvynol in combination with the entire array of drugs of abuse depending on the needs of a clinical program. Procedure II involves the principle of liquid-liquid extraction using either raw or spent urines.


Effect of 3 months' nasal steroid therapy on nasal T cells and Langerhans cells in patients suffering from allergic rhinitis.
The effect of nasal corticosteroid therapy on allergic rhinitis is uncertain. In a double-blind, placebo-controlled study over 3 months, we investigated the influence of a new corticosteroid spray, fluticasone propionate aqueous nasal spray (FPANS), on Langerhans cells (CD1a+ cells), HLA-DR+ cells, and T cells in nasal mucosa. Efficacy was evaluated by nasal symptom score. This treatment significantly decreased the number of CD1a+ cells and HLA-DR+ cells in the nasal mucosa. Furthermore, a clear trend of decreasing numbers of T cells in nasal epithelium was found. No change in nasal symptom score was found after the treatment period. These findings suggest that fluticasone propionate aqueous nasal spray decreases the antigen presentation in nasal allergy.


The role of rehearsal and reminding in the recall of categorized word lists.
Most theories of free recall emphasize the importance of retrieval in explaining temporal and semantic regularities in recall; rehearsal mechanisms are often absent or limit rehearsal to a subset of what was last rehearsed. However, in three experiments using the overt rehearsal method, we show clear evidence that just-presented items act as retrieval cues during encoding (study-phase retrieval) with prior related items rehearsed despite well over a dozen intervening items. Experiment 1 examined free recall of categorized and uncategorized lists of 32 words. In Experiments 2 and 3, we presented categorized lists of 24, 48, and 64 words for free recall or cued recall, with the category exemplars blocked in successive list positions (Experiment 2) or randomized throughout the list (Experiment 3). The probability of rehearsing a prior word was affected by its semantic similarity to the just-presented item, and the frequency and recency of its prior rehearsals. These rehearsal data suggest alternative interpretations to well-known recall phenomena. With randomized designs, the serial position curves were reinterpreted by when words were last rehearsed (which contributed to the list length effects), and semantic clustering and temporal contiguity effects at output were reinterpreted by whether words were co-rehearsed during study. The contrast with the blocked designs suggests that recall is sensitive to the relative (not absolute) recency of targeted list items. We discuss the benefits of incorporating rehearsal machinery into computational models of episodic memory, and suggest that the same retrieval processes that generate the recalls are used to generate the rehearsals.


Phylogeny of gracillariid leaf-mining moths: evolution of larval behaviour inferred from phylogenomic and Sanger data.
Gracillariidae is the most taxonomically diverse cosmopolitan leaf-mining moth family, consisting of nearly 2000 named species in 105 described genera, classified into eight extant subfamilies. The majority of gracillariid species are internal plant feeders as larvae, creating mines and galls in plant tissue. Despite their diversity and ecological adaptations, their phylogenetic relationships, especially among subfamilies, remain uncertain. Genomic data (83 taxa, 589 loci) were integrated with Sanger data (130 taxa, 22 loci), to reconstruct a phylogeny of Gracillariidae. Based on analyses of both datasets combined and analyzed separately, monophyly of Gracillariidae and all its subfamilies, monophyly of the clade "LAMPO" (subfamilies: Lithocolletinae, Acrocercopinae, Marmarinae, Phyllocnistinae, and Oecophyllembiinae) and relationships of its subclade "AMO" (subfamilies: Acrocercopinae, Marmarinae, and Oecophyllembiinae) were strongly supported. A sister-group relationship of Ornixolinae to the remainder of the family, and a monophyletic leaf roller lineage (Callicercops Vári + Parornichinae) + Gracillariinae, as sister to the "LAMPO" clade were supported by the most likely tree. Dating analyses indicate a mid-Cretaceous (105.3 Ma) origin of the family, followed by a rapid diversification into the nine subfamilies predating the Cretaceous-Palaeogene extinction. We hypothesize that advanced larval behaviours, such as making keeled or tentiform blotch mines, rolling leaves and galling, allowed gracillariids to better avoid larval parasitoids allowing them to further diversify. Finally, we stabilize the classification by formally re-establishing the subfamily ranks of Marmarinae stat.rev., Oecophyllembiinae stat.rev. and Parornichinae stat.rev., and erect a new subfamily, Callicercopinae Li, Ohshima and Kawahara to accommodate the enigmatic genus Callicercops.