Protective effect of gui zhi (Ramulus Cinnamomi) on abnormal levels of four amino acid neurotransmitters by chronically ma huang (Herb Ephedra) intoxicated prefrontal cortex in rats treated with a ma huang-gui zhi herb pair.
The Herb Ephedra (Ma Huang in Chinese)-Ramulus Cinnamomi (Gui Zhi in Chinese) herb pair is a classic traditional Chinese herb pair used to treat asthma, nose and lung congestion, and fever with anhidrosis. In previous study, we found that chronic administration of ma huang induced obvious neurodegeneration in rat brains, with the prefrontal cortex showing the greatest effect. Gui zhi decreased hyperactivity produced by repeated ma huang administration, and attenuated oxidative stress in rat prefrontal cortex induced by ma huang. The study was aimed to investigate the protective effect of gui zhi on ma huang-induced abnormal levels of four amino acid neurotransmitters in rat prefrontal cortex. All ma huang and ma huang-gui zhi herb pair extracts were prepared using methods of traditional Chinese medicine and were normalized based on the ephedrine content. Two-month-old male Sprague-Dawley rats (6 rats/group) were administered ma huang or ma huang-gui zhi herb pair extracts for 1, 3, 5 or 7 days (ephedrine = 48 mg/kg). The contents of ephedrine, glutamate (Glu), aspartic acid (Asp), glycine (Gly), and gamma-aminobutyric acid (GABA) in the prefrontal cortex were determined using ultra-high-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) at 0.5, 1.0, 5.0 h after administration. Ma huang significantly enhanced the levels of GABA, Gly, Glu and Asp in the prefrontal cortex, while gui zhi partially abolished the effects. Ma huang-induced neurotoxicity may be associated with its effects on amino acid neurotransmitters. Gui zhi is a promising neuroprotective agent against for ma huang-induced neurotoxicity. The information presented in this study will help supplement the available data for future ma huang-gui zhi herb pair compatibility studies.


Palbociclib use with grade 3 neutropenia in hormone receptor-positive metastatic breast cancer.
Neutropenia is the most common toxicity of CDK4/6 inhibitors, causing frequent dose interruptions. However, CDK4/6 inhibitor-induced neutropenia shows a benign clinical course in contrast to that caused by chemotherapy. Here, we investigated the safety of a new dose scheme for palbociclib, which avoids dose delays or reductions due to afebrile grade 3 neutropenia. A consecutive cohort of ER( +)/HER2( -) advanced breast cancer patients who received palbociclib between 2017 and 2018 was analyzed. The patients were classified into Group 1 (patients who maintained palbociclib dose with afebrile grade 3 neutropenia), Group 2 (patients who experienced any dose modification with afebrile grade 3 neutropenia), and Group 3 (patients without afebrile grade 3 neutropenia). The primary endpoint was febrile neutropenia incidence; other toxicities were compared with those of the PALOMA-2 trial. Among the 107 patients, 54.2%, 22.4%, and 23.4% were classified into Groups 1, 2, and 3, respectively. There was no febrile neutropenia in Groups 1 and 2 during palbociclib treatment. Group 1 showed higher incidence of thrombocytopenia (all-grade, 32.8%; grade 3-4, 8.6%) than Group 2 and the PALOMA-2 data, but there was no bleeding related to thrombocytopenia. Group 1 showed higher incidence of all-grade non-hematologic adverse events than Group 2; only one grade 3 non-hematologic toxicity was observed in Group 1. There were no treatment-related hospitalizations or deaths in Group 1. Thus, omitting palbociclib dose modification with afebrile grade 3 neutropenia is safe and tolerable without febrile neutropenia events. This scheme could be useful to avoid unnecessary reductions in palbociclib doses in future practice.


Dependence of 2,3-DPG and oxygen affinity of haemoglobin on sex and pregnancy in the guinea-pig.
Oxygen half-saturation of blood (P50), 2,3-diphosphoglycerate concentration (2,3-DPG) and Bohr effect were determined in male, and nonpregnant and pregnant female guinea pigs, according to a randomized block design. P50 was significantly higher in the female group (26.3 Torr plus or minus 0.22 SEM) than in the male group (24.8 Torr plus or minus 0.26 SEM) and was significantly lower in both these groups than in the pregnant group (27 Torr plus or minus 0.35 SEM). This difference in oxygen affinity was explained by differences in 2,3-DPG: 1.08 plus or minus 0.02 SEM in males, 1.24 plus or minus 0.03 in non-pregnant females and 1.34 plus or minus 0.03 mol/mol HB in pregnant females P50, 2,3-DPG and haemoglobin concentrations were significantly correlated for the ensemble of the 3 groups. There was no significant difference in Bohr effect between the 3 groups.


Amniotic fluid stem cell-based models to study the effects of gene mutations and toxicants on male germ cell formation.
Male infertility is a major public health issue predominantly caused by defects in germ cell development. In the past, studies on the genetic regulation of spermatogenesis as well as on negative environmental impacts have been hampered by the fact that human germ cell development is intractable to direct analysis in vivo. Compared with model organisms including mice, there are fundamental differences in the molecular processes of human germ cell development. Therefore, an in vitro model mimicking human sperm formation would be an extremely valuable research tool. In the recent past, both human embryonic stem (ES) cells and induced pluripotent stem (iPS) cells have been reported to harbour the potential to differentiate into primordial germ cells and gametes. We here discuss the possibility to use human amniotic fluid stem (AFS) cells as a biological model. Since their discovery in 2003, AFS cells have been characterized to differentiate into cells of all three germ layers, to be genomically stable, to have a high proliferative potential and to be non-tumourigenic. In addition, AFS cells are not subject of ethical concerns. In contrast to iPS cells, AFSs cells do not need ectopic induction of pluripotency, which is often associated with only imperfectly cleared epigenetic memory of the source cells. Since AFS cells can be derived from amniocentesis with disease-causing mutations and can be transfected with high efficiency, they could be used in probing gene functions for spermatogenesis and in screening for male reproductive toxicity.


A simple, effective method to create laparoscopic renal protective hypothermia with cold saline surface irrigation: clinical application and assessment.
We previously described the use of cold saline surface irrigation to achieve protective renal hypothermia in a laparoscopic partial nephrectomy porcine model. We now present our clinical application of this technique and characterization of the hypothermic effect during laparoscopic partial nephrectomy. Seven patients underwent elective laparoscopic partial nephrectomy augmented with our hypothermia technique. Parenchymal temperature sensors were placed to confirm cooling efficacy and efficiency. After transperitoneal exposure of the kidney we performed temporary hilar vascular occlusion. Surface cooling with almost freezing normal saline was delivered with a laparoscopic suction/irrigation device. Tumor laparoscopic resection and renal reconstruction were completed. Outcome measures included intraoperative changes with hypothermia and postoperative estimated glomerular filtration rate changes. All patients successfully underwent laparoscopic partial nephrectomy without complications or evidence of residual disease. A protective renal parenchymal temperature of less than 20C was achieved at a mean application time of 8.3 minutes. The hypothermic window of 15C to 25C was maintained an average of 30.4 minutes. In 2 cases cooling was repeated and 4 minutes were required to lower the temperature below 20C. The overall mean core body temperature decrease was 1.28C. At a mean followup of 22.4 months the median preoperative, immediate postoperative and final estimated glomerular filtration rate was 75, 65 and 71 ml/minute/1.73 m(2), respectively. There was no evidence of disease recurrence on followup imaging. Our technique involving cold saline surface irrigation to achieve protective renal hypothermia is reproducible, and uses readily available laparoscopic instruments and equipment. This technique can be done simply and effectively, and may expand the use of laparoscopic partial nephrectomy.


Linking the human cytogenetic map with nucleotide sequence: the CCAP clone set.
We present the completed dataset and clone repository of the Cancer Chromosome Aberration Project (CCAP), an initiative developed and funded through the intramural program of the U.S. National Cancer Institute, to provide seamless linkage of human cytogenetic markers with the primary nucleotide sequence of the human genome. Spaced at 1-2 Mb intervals across the human genome, 1,339 bacterial artificial chromosome (BAC) clones have been localized to chromosomal bands through high-resolution fluorescence in situ hybridization (FISH) mapping. Of these clones, 99.8% can be positioned on the primary human genome sequence and 95% are placed at or close to their precise nucleotide starts and stops. This dataset can be studied and manipulated within generally available public Web sites. The clones are available from a commercial repository. The CCAP BAC clone set provides anchors for the interrogation of gene and sequence involvement in oncogenic and developmental disorders when the starting point is the recognition of a structural, numerical, or interstitial chromosomal aberration. This dataset also provides a current view of the quality and coherence of the available genome sequence and insight into the nucleotide and three-dimensional structures that manifest as Giemsa light and dark chromosomal banding patterns.


A new MAGE-4 antigenic peptide recognized by cytolytic T lymphocytes on HLA-A24 carcinoma cells.
"Cancer-germline" genes such as those of the MAGE family are expressed in many tumors and in male germline cells, but are silent in other normal tissues. They encode tumor specific antigens that are used in cancer immunotherapy trials. MAGE-4 antigens represent promising targets for cancer immunotherapy because gene MAGE-4 is expressed in more than 50% of carcinomas of the esophagus, lung, bladder, and head and neck. To identify new MAGE-4 antigenic peptides, we have folded HLA-A*2402 soluble molecules with candidate peptide NYKRCFPVI, which corresponds to amino acids 143 to151 of the MAGE-4 protein. A24/MAGE-4 multimers were used to isolate a cytolytic T cell clone that recognized the MAGE-4 peptide from the blood cells of a donor without cancer. This clone lysed specifically A24 carcinoma cells expressing MAGE-4. The antigenic peptide is processed more efficiently in tumor cells pre-treated with IFN-gamma. This MAGE-4 peptide could represent an interesting target for immunotherapy because it is presented by HLA-A24 molecules, which are widely expressed in different ethnic groups.


Bifid scrotum, perineal hamartoma and high imperforate anus: a case report.
This is a case report of a newborn patient with imperforate anus, urethro-colonic fistula, perianal hamartoma, and bifid scrotum. Successful staged repair of these anomalies is described together with review of the embriology related to the case.


The evolution of base composition and phylogenetic inference.
Base composition varies at all levels of the phylogenetic hierarchy and throughout the genome, and can be caused by active selection or passive mutation pressure. This variation can make reconstructing trees difficult. However, recent tree-based analyses have shed light on the forces responsible for the evolution of base composition, forces that might be very general. More explicit tree-based work is encouraged.


Genomewide molecular and biologic characterization of biomembrane formation adjacent to a methacrylate spacer in the rat femoral segmental defect model.
This study focuses upon the morphologic and molecular features of the layer of cells, termed the "biomembrane," which forms around methacrylate spacers in bone segmental defects. The objective of this research was to assess the biomembrane formed in a novel rodent femoral segmental defect model at 4, 8, and 16 weeks with histologic and molecular studies. Following Institutional Animal Care and Use Committee approval, a segmental defect was created in the rat femur and stabilized with the AO LockingRatNail and analyzed at 4, 8, and 16 weeks postsurgery using digital radiologic imaging, morphological and immunohistochemical studies, and genomewide gene expression studies employing microarray analysis. The biomembrane formed around the methacrylate spacer was rich in vasculature, which showed vascular endothelial growth factor immunolocalization. The biomembrane supported development of foci of bone and cartilage within it. Bone morphogenetic protein 2 immunolocalization and gene expression were positive within developing osseous and chondrocyte foci. Microarray analysis showed significant expression of key genes related to bone and cartilage formation and angiogenesis. This rat bone model was effective in creation of the biomembrane. Bone and cartilage foci were formed within the vascularized biomembrane with associated expression of genes critical for bone and cartilage development/formation and vascularization. The polymethyl methacrylate-induced biomembrane offers an exciting potential solution for segmental defects; the biomembrane, may act as a receptive bed and also serve as a source for mesenchymal stem cells, which could be recruited/directed for the healing process.


FluClass: A novel algorithm and approach to score and visualize the phylogeny of the influenza virus using mass spectrometry.
A novel computer algorithm FluClass has been developed to facilitate the phylogenetic classification of influenza virus using mass spectral data. FluClass accepts a DNA or protein-based phylogenetic tree as input and generates theoretical peptide mass lists for each node. An experimental mass spectrum from an influenza virus protein digest is then placed onto the phylogenetic tree using a novel random resampling function (Z-score) that allows the scoring of spectrum against both internal and leaf nodes. Testing of the algorithm using hemagglutinin protein sequences from human-host influenza viruses showed that the Z-score performs comparably to the Profound scoring method for the scoring of leaf nodes and is substantially better at scoring internal nodes. Scoring of internal nodes allows colorizations of nodes of the phylogenetic tree enabling the classification of the query spectrum to be rapidly visualized. Finally we demonstrate the utility of FluClass on experimental spectra from six strains. Given that mass spectrometry data can be generated rapidly for influenza virus proteins, FluClass provides a fast and direct method for phylogenetic analysis of influenza proteins.


Economic evaluation of programs against COVID-19: A systematic review.
The COVID-19 pandemic has become a public health emergency and raised global concerns in about 213 countries without vaccines and with limited medical capacity to treat the disease. The COVID-19 has prompted an urgent search for effective interventions, and there is little information about the money value of treatments. The present study aimed to summarize economic evaluation evidence of preventing strategies, programs, and treatments of COVID-19. We searched Medline/PubMed, Cochrane Library, Web of Science Core Collection, Embase, Scopus, Google Scholar, and specialized databases of economic evaluation from December 2019 to July 2020 to identify relevant literature to economic evaluation of programs against COVID-19. Two researchers screened titles and abstracts, extracted data from full-text articles, and did their quality assessment by the Consolidated Health Economic Evaluation Reporting Standards (CHEERS) checklist. Then, quality synthesis of results was done. Twenty-six studies of economic evaluations met our inclusion criteria. The CHEERS scores for most studies (n = 9) were 85 or higher (excellent quality). Eight studies scored 70 to 85 (good quality), eight studies scored 55 to 70 (average quality), and one study < %55 (poor quality). The decision-analytic modeling was applied to twenty-three studies (88%) to evaluate their services. Most studies utilized the SIR model for outcomes. In studies with long-time horizons, social distancing was more cost-effective than quarantine, non-intervention, and herd immunity. Personal protective equipment was more cost-effective in the short-term than non-intervention. Screening tests were cost-effective in all studies. The results suggested screening tests and social distancing to be cost-effective alternatives in preventing and controlling COVID-19 on a long-time horizon. However, evidence is still insufficient and too heterogeneous to allow any definite conclusions regarding costs of interventions. Further research as are required in the future.


Maternal satisfaction with breastfeeding in the first month postpartum and associated factors.
Breastfeeding success has been measured based on its duration, disregarding satisfaction with the maternal experience. Studies to investigate maternal satisfaction with breastfeeding are rare, especially in Brazil, and little is known about their determinants. The aim of this study was to measure the level of satisfaction with breastfeeding in a group of women in the first month of their child's life, and to identify factors associated with higher maternal satisfaction. A cross-sectional study nested within a cohort was conducted with 287 women recruited at two (one public, one private) maternity services in the city of Porto Alegre, southern Brazil, from January to July 2016. Women residing in the municipality who had given birth to a healthy singleton born at term, were rooming in, and had initiated breastfeeding were randomly included. During the week after the child was 30 days old, women were interviewed at their homes to measure the level of maternal satisfaction with breastfeeding, using the Maternal Breastfeeding Evaluation Scale (MBFES), validated for use in the Brazilian population. Associations between maternal satisfaction and explanatory variables were estimated using multivariate Poisson regression with robust variance in a four-level hierarchical approach. Satisfaction level was categorized using as cutoff point the median score obtained with the MBFES. Women with scores equal to or above the median were considered to have higher levels of satisfaction, whereas those scoring below the median were considered to be less satisfied. Maternal satisfaction with breastfeeding in the first month postpartum was high, with a median score of 124 on MBFES, close to the maximum score (145 points). The prevalence of more elevated levels of satisfaction with breastfeeding was higher among women with brown (pardo) and black skin color (prevalence ratio [PR] 1.33, 95%CI 1.05;1.69), those who lived with the partner (PR 1.75, 95%CI 1.05;2.94), who planned to breastfeed for 12 months or more (PR 1.48, 95%CI 1.02;2.17), and who did not report low milk supply (PR 1.47, 95%CI 1.03;2.10) or cracked nipples (PR 1.29, 95%CI 1.01;1.65). The factors associated with maternal satisfaction with breastfeeding in the first month postpartum include individual factors and maternal expectations, family constitution, as well as breastfeeding-related problems.


Early and long-term effects of acute caloric deprivation in obese diabetic patients.
It is generally assumed that diet therapy can ameliorate the metabolic derangements experienced by obese type 2 diabetic patients, thereby leading to discontinuation of insulin or oral sulfonylurea drug therapy. We decided to retrospectively investigate which clinical and biochemical parameters affect therapeutic responses. Sixty-four poorly controlled obese diabetic patients were hospitalized and placed on a precisely defined, hypocaloric diet. Known duration of diabetes, type of pharmacologic therapy, body weight, weight loss, fasting plasma glucose concentrations, C-peptide levels, hemoglobin A1C, and plasma lipid levels were assessed, as were nitrogen and electrolyte balances. Average weight loss was 13 pounds in a mean of 23 days. During hospitalization, the mean fasting plasma glucose value for the group fell from 221 +/- 10 to 122 +/- 5 mg/dl. In 45 patients (73 percent), the final fasting plasma glucose level was less than 125 mg/dl (mean: 102 +/- 2 mg/dl). Oral glucose tolerance even in those patients in whom fasting plasma glucose levels normalized was still grossly diabetic at the end of the hospital stay, deteriorating further after three days of liberalized caloric intake. In part this may have been due to decreased insulin secretory reserve as reflected by blunted plasma C-peptide response. Forty of 42 patients who entered the study taking insulin were able to discontinue the drug within one to seven days of hospitalization. After a mean follow-up period of 19 months, only 10 of 50 patients continued to maintain fasting euglycemia; five were on diet alone, and five were receiving oral hypoglycemic agents. Thirteen patients were receiving insulin therapy. Diet therapy in these patients resulted in short-term improvement of glycemic control and, in the majority, normalization of fasting plasma glucose levels. However, long-term outpatient follow-up revealed that relapse occurred in most patients.


Prophylactic HPV vaccines: reducing the burden of HPV-related diseases.
HPV-associated diseases, such as cervical and other anogenital cancers, cervical and anal intraepithelial neoplasia, genital warts, and recurrent respiratory papillomatosis confer considerable morbidity and mortality, and are significant health care concerns. Successful vaccination strategies that protect against HPV infection are expected to substantially reduce HPV-related disease burden. Prophylactic HPV vaccines in late stages of clinical testing are composed of HPV L1 capsid protein that self-assemble into virus-like particles (VLPs) when expressed in recombinant systems. Proof-of-principle trials have suggested that intramuscular injections of VLPs results in strong adaptive immune responses, both B- and T-cell mediated, that are capable of neutralizing subsequent natural infections. Furthermore, phase 2 trials of a bivalent vaccine designed to protect against high-risk HPV types 16 and 18 and a quadrivalent vaccine designed to protect against HPV 16 and 18, and low-risk, genital wart-causing HPV 6 and 11 have demonstrated that VLP vaccines reduce the incidence of HPV-associated disease in vaccinated individuals. To derive the greatest public health benefit, HPV vaccines offering protection from cervical cancer and genital warts will, ideally, be administered prior to the initiation of sexual activity; therefore, educational initiatives will be essential to communicate the risks and adverse consequences of HPV infection and to foster widespread vaccine acceptance.


Rational Construction of Electrically Conductive Covalent Organic Frameworks through Encapsulating Fullerene via Donor-Acceptor Interaction.
A new kind of perylene-based 2D covalent organic framework (COF) is designed and synthesized based on the C2 + C2 topological diagram. The perylene-based COF is constructed via the condensation reaction using 2,5,8,11-tetrakis((4-formylphenyl) perylene (TFPPer) and 2,5,8,11-tetrakis(4-aminophenyl) perylene (TAPPer) as building blocks. The resulting TFPPer-TAPPer-COF features high crystallinity, excellent stability, intrinsic porosity, and an electron-rich skeleton. Significantly, the electrical performance of the COF can be enhanced through the encapsulation of fullerene (C60 ) into the 1D channels via donor-acceptor interaction. Compared to the pristine COF, the electrical conductivity of C60 @TFPPer-TAPPer-COF can be greatly increased from 8.98 × 10-8 to 1.59 × 10-5 S cm-1 , meanwhile the carrier mobility rises from 1.04 × 10-3 to 4.23 × 10-2 cm2 V-1 s-1 . The improvement in electrical performance stems from the strong donor-acceptor interaction between perylene and C60 . These results provide insights into the rational construction of conductive COFs through donor-acceptor interaction and demonstrate their great potential in related application fields.


Assessments of prolonged effects of desflurane and sevoflurane on motor learning deficits in aged AppNL-G-F/NL-G-F mice.
As the proportion of elderly in society increases, so do the number of older patients undergoing surgical procedures. This is concerning as exposure to anesthesia has been identified as a risk factor for Alzheimer's disease (AD). However, the causal relationship between clinical AD development and anesthesia remains conjectural. Preclinical studies have demonstrated that anesthesia, such as halothane, isoflurane, and sevoflurane, induces AD-like pathophysiological changes and cognitive impairments in transgenic mouse models of AD. Desflurane does not have these effects and is expected to have more potential for use in elderly patients, yet little is known about its effects, especially on non-cognitive functions, such as motor and emotional functions. Thus, we examined the postanesthetic effects of desflurane and sevoflurane on motor and emotional function in aged AppNL-G-F/NL-G-F (App-KI) mice. This is a recently developed transgenic mouse model of AD exhibiting amyloid β peptide (Aβ) amyloidosis and a neuroinflammatory response in an age-dependent manner without non-physiological amyloid precursor protein (APP) overexpression. Mice were subjected to a short behavioral test battery consisting of an elevated plus maze, a balance beam test, and a tail suspension test seven days after exposure to 8.0% desflurane for 6 h or 2.8% sevoflurane for 2 h. App-KI mice showed significant increments in the percentage of entry and time spent in open arms in the elevated plus maze, increments in the number of slips and latency to traverse for the balance beam test, increments in the limb clasping score, increments in immobile duration, and decrements in latency to first immobile episode for the tail suspension test compared to age-matched wild type (WT) controls. Desflurane- and sevoflurane-exposed App-KI mice showed a delayed decrement in the number of slips for each trial in the balance beam test, while air-treated App-KI mice rapidly improved their performance, and increased their clasping behavior in the tail suspension test. Furthermore, App-KI inhibited the change in membrane GluA3 following exposure to anesthetics in the cerebellum. These results suggest high validity of App-KI mice as an animal model of AD.


Graded exercise therapy does not restore the ability to work in ME/CFS - Rethinking of a Cochrane review.
Cochrane recently amended its exercise review for Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) in response to an official complaint. To determine if the amended review has addressed the concerns raised about the previous review and if exercise is an effective treatment that restores the ability to work in ME/CFS. The authors reviewed the amended Cochrane exercise review and the eight trials in it by paying particular interest to the objective outcomes. We also summarised the recently published review of work rehabilitation and medical retirement for ME/CFS. The Cochrane review concluded that graded exercise therapy (GET) improves fatigue at the end of treatment compared to no-treatment. However, the review did not consider the unreliability of subjective outcomes in non-blinded trials, the objective outcomes which showed that GET is not effective, or the serious flaws of the studies included in the review. These flaws included badly matched control groups, relying on an unreliable fatigue instrument as primary outcome, outcome switching, p-hacking, ignoring evidence of harms, etc. The review did also not take into account that GET does not restore the ability to work. GET not only fails to objectively improve function significantly or to restore the ability to work, but it is also detrimental to the health of≥50% of patients, according to a multitude of patient surveys. Consequently, it should not be recommended.


Incidence and treatment of candida esophagitis in patients undergoing renal transplantation. Data from the Minnesota prospective randomized trial of cyclosporine versus antilymphocyte globulin-azathioprine.
Of 224 consecutive renal transplant patients in a prospective, randomized immunosuppressive trial, candida esophagitis developed in 5 despite nystatin prophylaxis. No differences were noted between cyclosporine and antilymphocyte globulin-azathioprine immunosuppressive treatment. All patients were diabetic, and four were recipients of cadaver kidneys. Candida esophagitis occurred within 6 months after transplantation, and only one patient had recurrence. All patients responded to treatment consisting of 2 to 6 days of intravenous amphotericin B (0.2 to 2 mg/kg total dose). The prevalence of candida esophagitis was not related to rejection episodes. Three of five patients eventually died, one 2 weeks after resolution of candida esophagitis from a hypoglycemic episode, one from acute exacerbation of pulmonary failure and relapsing pancreatitis in association with candida esophagitis and therapy-resistant candidemia, and one 17 months after candida esophagitis from pulmonary edema. Our findings show that candida esophagitis by itself is an easily managed complication, but is also a sign of potentially increased morbidity in these patients.


Results of multiple ligament reconstruction after knee dislocation--A prospective study with 95 patients and minimum 2-year follow up.
There is still a lack of clinical data in arthroscopic treatment for treating multiple ligament injuries. This study aims to evaluate the clinical outcomes of patients with multiple ligament injuries undergoing treatment based on the classification stage and type of injury. A prospective, clinical trial on multiple ligament injuries was planned, which included 95 patients (58 men and 37 women; age: 42.8 ± 11.9 [range, 18-63] years) from October 2017 to June 2018. Injuries were classified into three stages (emergency stage < 24 h; acute stage: 24 h to 3 weeks, and chronic stage: > 3 weeks) and six types (KD I-VI) based on injuries time and structures, which indicated appropriate treatments. The clinical outcomes were evaluated at 2, 4, 6, 8, and 12 weeks and at 6, 9, 12 months and 24 months after surgery. The final choices in efficacy index included International Knee Documentation Committee (IKDC) score, Lysholm score, visual analog scale (VAS) score, and range of motion. During the follow-up, all patients exhibited statistically significant functional improvement in the injured limb compared with their preoperative situation. The mean postoperative scores of acute stage patients at 2-year follow-up were IKDC subjective score, 77.54 ± 11.53; Lysholm score, 85.96 ± 9.39; Tegner score, 4.13 ± 1.08; and VAS score, 1.21 ± 0.76. The mean postoperative scores of chronic stage patents at 2-year follow-up were IKDC subjective score, 74.61 ± 12.38; Lysholm score, 81.71 ± 10.80; Tegner score, 3.96 ± 1.14; and VAS score, 1.71 ± 0.60. The IKDC subjective score, Lysholm score, and Tegner score were significantly improved (P < 0.01) and the VAS score was significantly decreased (P < 0.01) at 2-year follow-up. Regarding the multiple ligament injuries classification, patients with more structural damage in stages V and VI showed less progress in functional recovery than those in stages I-IV. This new classification with three stages and six types helps to identify the severity of injury and plan the management effectively. The outcomes were encouraging and the subjective functional results showed significant improvement at 2-year follow-up. Prospective clinical trial. II.


Distinct comammox Nitrospira catalyze ammonia oxidation in a full-scale groundwater treatment bioreactor under copper limited conditions.
Microbial ammonia oxidation is the initial nitrification step used in biological nitrogen-removal during water treatment processes, and the discovery of complete ammonia-oxidizing (comammox) bacteria added a novel member to this functional group. It is important to identify and understand the predominant microorganisms responsible for ammonium removal in biotechnological process design and optimization. In this study, we used a full-scale bioreactor to treat ammonium in groundwater (9.3 ± 0.5 mg NH4+-N/L) and investigated the key ammonia-oxidizing prokaryotes present. The groundwater ammonium was stably and efficiently oxidized throughout ∼700 days of bioreactor operation. 16S rRNA gene amplicon sequencing of the bioreactor community showed a high abundance of Nitrospira (12.5-45.9%), with the dominant sequence variant (3.5-37.8%) most closely related to Candidatus Nitrospira nitrosa. Furthermore, analyses of amoA, the marker gene for ammonia oxidation, indicated the presence of two distinct comammox Nitrospira populations, however, the relative abundance of only one of these populations was strongly correlated to ammonia oxidation rates and was robustly expressed. After 380 days of operation copper wires were immersed into the reactor at 0.04-0.06 m2/m3 tank, which caused a gradual abundance increase of one discrete comammox Nitrospira population. However, further increase of the copper dosing (0.08 m2/m3 tank) inverted the most abundant ammonia-oxidizing population to Nitrosomonas sp. These results indicate that comammox Nitrospira were capable of efficient ammonium removal in groundwater without exogenous nutrients, but copper addition can stimulate comammox Nitrospira or lead to dominance of Nitrosomonas depending on dosage.


Optogenetic control of endogenous Ca(2+) channels in vivo.
Calcium (Ca(2+)) signals that are precisely modulated in space and time mediate a myriad of cellular processes, including contraction, excitation, growth, differentiation and apoptosis. However, study of Ca(2+) responses has been hampered by technological limitations of existing Ca(2+)-modulating tools. Here we present OptoSTIM1, an optogenetic tool for manipulating intracellular Ca(2+) levels through activation of Ca(2+)-selective endogenous Ca(2+) release-activated Ca(2+) (CRAC) channels. Using OptoSTIM1, which combines a plant photoreceptor and the CRAC channel regulator STIM1 (ref. 4), we quantitatively and qualitatively controlled intracellular Ca(2+) levels in various biological systems, including zebrafish embryos and human embryonic stem cells. We demonstrate that activating OptoSTIM1 in the CA1 hippocampal region of mice selectively reinforced contextual memory formation. The broad utility of OptoSTIM1 will expand our mechanistic understanding of numerous Ca(2+)-associated processes and facilitate screening for drug candidates that antagonize Ca(2+) signals.


Experience dependence of neural responses to different classes of male songs in the primary auditory forebrain of female songbirds.
There is both extensive species-specificity and critical experience-dependence in the recognition of own species songs in many songbird species. For example, female zebra finches Taeniopygia guttata raised by their parents show behavioral preferences for the songs of the father over unfamiliar conspecific males and for unfamiliar songs of conspecifics over heterospecifics. Behavioral discrimination between different species' songs is also displayed by females raised without exposure to any male songs but it is diminished in females raised by heterospecific foster parents. We tested whether neural responses in the female auditory forebrain paralleled each of these known behavioral patterns in song-class discrimination. We analyzed spike rates, above background levels, recorded from single units in the L2a subregion of the field L complex of female zebra finches. In subjects raised by genetic parents, spike rates were similar to songs of fathers and unfamiliar male zebra finches, and higher to unfamiliar conspecific over unfamiliar heterospecific songs. In females raised in isolation from male songs, we also found higher spike rates to unfamiliar conspecific over heterospecific songs. In females raised by heterospecific foster parents, spike rates were similar in response to songs of the foster father and unfamiliar males of the foster species, similar between unfamiliar songs of conspecifics and the heterospecific foster species, and higher to unfamiliar songs of the foster species over a third finch species. Thus, in parallel to the experience-dependence of females' behaviors in response to different male song classes, differences in social experiences can also alter neural response patterns to male song classes in the auditory forebrain of female zebra finches.


Orgasm-induced prolactin secretion: feedback control of sexual drive?
Recent studies from our laboratory have investigated the hormonal response to various forms of sexual stimulation, including film, masturbation, and coitus in both men and women. This series of studies clearly demonstrated that plasma prolactin (PRL) concentrations are substantially increased for over 1h following orgasm (masturbation and coitus conditions) in both men and women, but unchanged following sexual arousal without orgasm. Here we discuss evidence suggesting that the PRL response to orgasm may play an important role in the control of acute sexual arousal following orgasm. Supporting this position, chronic elevations of PRL (hyperprolactinemia) produce pronounced reductions in animal sexual activity, and significant reduction of libido and gonadal function in both men and women. These data suggest that PRL may represent a peripheral regulatory factor for reproductive function, and/or a feedback mechanism that signals CNS centres controlling sexual arousal and behaviour. Thus, we propose a theoretical model of the role of PRL as a neuroendocrine reproductive reflex.


Nest predators and the evolution of avian reproductive strategies: a comparison of Australian and New Zealand birds.
Nest predation has been considered an important factor in the evolution of avian life histories: smaller clutches and shorter incubation and nestling periods are expected where nest predation has significant effects on reproductive success. Unlike the Australian avifauna, terrestrial New Zealand birds have evolved in the absence of reptilian and mammalian predators. Here we compare the reproductive strategies of terrestrial native New Zealand birds with those of their Australian sister taxa. In 11 of 14 comparisons, New Zealand birds were larger than their Australian relatives, but we did not find any significant differences in reproductive tactics between the two regions, a result inconsistent with the nest predation hypothesis. We discuss several reasons why this may be so. One possibility is that selection imposed on avian life history tactics by mammalian predators following the arrival of humans in New Zealand has led to strategies similar to those adopted in Australia.


Amino acids and their derivatives mediating defense priming and growth tradeoff.
Plant response to pathogens attacks generally comes at the expense of growth. Defense priming is widely accepted as an efficient strategy used for augmenting resistance with reduced fitness in terms of growth and yield. Plant-derived small molecules, both primary as well as secondary metabolites, can function as activators to prime plant defense. Amino acids and their derivatives regulate numerous aspects of plant growth and development, and biotic and abiotic stress responses. In this review, we discuss the recent progress in understanding the roles of amino acids and related molecules in defense priming and their link with plant growth. We also highlight some of the outstanding questions and provide an outlook on the prospects of 'engineering' the tradeoff between defense and growth in plants.


Stent implantation in the superficial femoral artery: short thrombelastometry-derived coagulation times identify patients with late in-stent restenosis.
The mechanisms of restenosis, the recurrence of luminal narrowing, are complex and incompletely understood to date. Thrombin, the pivotal enzyme in haemostasis, presumably contributes to the formation of in-stent restenosis (ISR). It was therefore the aim of our study to investigate whether blood coagulation/thrombin generation plays a critical role in the formation of ISR in peripheral artery disease patients with stent angioplasty in the superficial femoral artery. We aimed to examine in this retrospective study whether patients with high-degree restenosis (50-75% lumen diameter reduction, n=20) are in a hypercoaguable state implying enhanced readiness to generate thrombin compared to patients with low-degree restenosis (<50% lumen diameter reduction, n=14). The coagulation tests calibrated automated thrombography, activated partial thromboplastin time, platelet aggregation, platelet adhesion, fibrinogen, and microparticles' procoagulant activity did not indicate a different coagulation status in the two patient groups. However, the thrombelastometry-derived value Coagulation Time (CT) was significantly shorter in the high-degree restenosis group (p=0.012), indicating a hypercoagulable state of patients with high-degree restenosis. Under our experimental conditions, CTs shorter than 444.5s identify patients at high risk (sensitivity=95%) for luminal narrowing. Our study supports the assumption that blood coagulation/thrombin generation plays a critical role in the development of ISR in peripheral arteries after stent insertion and that the thrombelastometry-derived CT might be a suitable value to identify peripheral artery disease patients at risk for development of high-degree in-stent restenosis in the superficial femoral artery.


Acute induction of joint inflammation in the rat by poly I . poly C.
Intraarticular injection of interferon inducer, double-stranded polyinosinate-polycytidylate (Poly I . C) caused acute synovitis in rats. This acute inflammatory response was accompanied by an increased concentration of prostaglandin E (PGE) in the synovial tissue. Double-stranded polyadenylate-polyuridylate (Poly A . Poly U) was less potent than Poly I . Poly C in inducing synovitis and increasing PGE concentration, while single-stranded polyinosinate (Poly I) or polycytidylate (Poly C) were inactive in these respects. Intraarticular injection of partially purified mouse fibroblast interferon also induced synovial inflammation. The present results suggest that interferon may be a mediator of viral inflammatory responses.


Standardization of tritiated water and 204Tl by TDCR liquid scintillation counting.
The triple to double coincidence ratio method was used with a combined uncertainty of 1-2% to standardize tritiated water and a solution of 204Tl. The vial with liquid scintillator, in which the sample to be measured was dissolved, is optically coupled to three photomultipliers. The electronic module MAC-3 assures the selection of double and triple coincidences count rates, D and T, from the three counting channels. It contains the gating circuits, necessary to obtain the livetime value and the extended deadtime circuit. Specific computer programs were used to calculate the free parameter value, the efficiency of D and so, the value of the activity. The optimal value of the Birks ionisation-quenching parameter, kB, was evaluated by changing the detection efficiency with grey filters. Three types of liquid scintillators, namely InstaGel, PPO+POPOP+Triton in toluene and Ultima Gold, were employed. 204Tl was measured in the frame of an international comparison organized by BIPM. For tritiated water a comparison was made with LNHB-Saclay; the relative difference between the obtained values for the massic activity was only 0.2%.


[Phagocytic lymphocyte-like elements (PLLE) and irradiation in vitro].
The peripheral blood of rats contains lymphocyte-like phagocytes after intracardial injection of India ink. In vitro irradiation of whole blood (1000 r - 60Co), causes marked reduction in whole lymphocytes but does not modify the number and morphological integrity of the phagocytes, which can therefore be held to be comparatively radioresistant: as such, they could belong either to the mononucleate phagocyte system or to the lymphatic system (T4 lymphocytes). Since T-lymphocytes are not capable of phagocytosis, the former hypothesis would appear the most probable.


[Juvenile fibromyalgia syndrome].
The aim was to develop a guideline for diagnostic procedures and treatment of juvenile fibromyalgia syndrome (JFMS) in cooperation with 10 German medical and psychological associations and 2 patient self-help groups. A systematic literature search, including all controlled studies evaluating diagnosis and treatment of JFMS, was performed in the Cochran Collaboration Reviews (1993-12/2006), Medline (1980-12/2006), PsychInfo (1966-12/2006) and Scopus (1980-12/2006). Levels of evidence were assigned according to the classification system of the Oxford Centre for Evidence-Based Medicine. Grading of the strengths of recommendations was performed according to the German program for disease management guidelines. Standardized procedures to reach a consensus on recommendations were used. Pain in children/adolescents involving several body areas and lasting >3 months without an obvious somatic cause is called JFMS or pain amplification syndrome. Therapeutically, a multidisciplinary concept with psychotherapy and physiotherapy, relaxation techniques and patient education is recommended. These guideline will contribute to a better recognition and standardized care of patients with JFMS and facilitate clinical studies.


Inhaled silica nanoparticles exacerbate atherosclerosis through skewing macrophage polarization towards M1 phenotype.
Exposure to environmental nanoparticles is related to the adverse impact on health, including cardiovascular system. Various forms of nanoparticles have been reported to interact with endothelium and induce inflammation. However, the potential role of nanoparticles in the pathogenesis of atherosclerosis and their mechanisms of action are still unclear. The aim of this study was to investigate the effect of two broadly used nanomaterials, which also occur in natural environment - silicon oxide (SiO2) and ferric oxide (Fe2O3) in the form of nanoparticles (NPs) - on the development of atherosclerosis. We used apolipoprotein E-knockout mice exposed to silica and ferric oxide nanoparticles in a whole body inhalation chamber. Inhaled silica nanoparticles augmented the atherosclerotic lesions and increased the percentage of pro-inflammatory M1 macrophages in both the plaque and the peritoneum in apoE-/- mice. Exposure to ferric oxide nanoparticles did not enhance atherogenesis process, however, it caused significant changes in the atherosclerotic plaque composition (elevated content of CD68-positive macrophages and enlarged necrotic core accompanied by the decreased level of M1 macrophages). Both silica and ferric oxide NPs altered the phenotype of T lymphocytes in the spleen by promoting polarization towards Th17 cells. Exposure to silica and ferric oxide nanoparticles exerts impact on atherosclerosis development and plaque composition. Pro-atherogenic abilities of silica nanoparticles are associated with activation of pro-inflammatory macrophages.


Human sparganosis, a neglected food borne zoonosis.
Human sparganosis is a food borne zoonosis caused by the plerocercoid larvae (spargana) of various diphyllobothroid tapeworms of the genus Spirometra. Human infections are acquired by ingesting the raw or undercooked meat of snakes or frogs, drinking untreated water, or using raw flesh in traditional poultices. More than 1600 cases of sparganosis have been documented worldwide, mostly in east and southeast Asia. Sporadic cases have been reported in South America, Europe, and Africa, and several cases have been described in travellers returning from endemic regions. Epidemiological data suggest that the increased effect of sparganosis on human health is because of greater consumption of raw meat of freshwater frogs and snakes. This Review provides information about the Spirometra parasites and their lifecycles, summarises clinical features, diagnosis, and treatment of human sparganosis, and describes geographical distribution and infection characteristics of Spirometra parasites in host animals.


Proteasome-dependent degradation of oxidized proteins in MRC-5 fibroblasts.
Fibroblasts were exposed to various concentrations of hydrogen peroxide and the removal of oxidized proteins was followed by determining protein-bound carbonyls. Fibroblasts are able to increase the turnover of metabolically radiolabeled proteins after treatment with hydrogen peroxide. It was demonstrated for the first time, that the increased protein turnover was accompanied by a removal of protein-bound carbonyl groups. The proteasome-specific inhibitor lactacystin was able to inhibit the elimination of protein-bound carbonyl groups. Therefore, the key role of the proteasome in the degradation of oxidized proteins in fibroblasts could be demonstrated.


Functional adaptive landscapes predict terrestrial capacity at the origin of limbs.
The acquisition of terrestrial, limb-based locomotion during tetrapod evolution has remained a subject of debate for more than a century1,2. Our current understanding of the locomotor transition from water to land is largely based on a few exemplar fossils such as Tiktaalik3, Acanthostega4, Ichthyostega5 and Pederpes6. However, isolated bony elements may reveal hidden functional diversity, providing a more comprehensive evolutionary perspective7. Here we analyse 40 three-dimensionally preserved humeri from extinct tetrapodomorphs that span the fin-to-limb transition and use functionally informed ecological adaptive landscapes8-10 to reconstruct the evolution of terrestrial locomotion. We show that evolutionary changes in the shape of the humerus are driven by ecology and phylogeny and are associated with functional trade-offs related to locomotor performance. Two divergent adaptive landscapes are recovered for aquatic fishes and terrestrial crown tetrapods, each of which is defined by a different combination of functional specializations. Humeri of stem tetrapods share a unique suite of functional adaptations, but do not conform to their own predicted adaptive peak. Instead, humeri of stem tetrapods fall at the base of the crown tetrapod landscape, indicating that the capacity for terrestrial locomotion occurred with the origin of limbs. Our results suggest that stem tetrapods may have used transitional gaits5,11 during the initial stages of land exploration, stabilized by the opposing selective pressures of their amphibious habits. Effective limb-based locomotion did not arise until loss of the ancestral 'L-shaped' humerus in the crown group, setting the stage for the diversification of terrestrial tetrapods and the establishment of modern ecological niches12,13.


[Noninvasive diagnosis of posttraumatic thoracic splenosis].
Thoracic splenosis is a rare occurrence that has usually been diagnosed by invasive procedures to allow a pathologic diagnosis to be reached. A firm diagnosis can now be made with the help of new, noninvasive imaging techniques. We report the case of a 34-year-old man with a history of severe thoracic-abdominal injury, including rupture of the spleen and left diaphragm. During computed tomography of the thorax related to a different diagnosis, nonspecific nodules were observed, although the patient was asymptomatic. A suspected diagnosis of thoracic splenosis was confirmed by technetium-99 sulfur colloid scintigraphy.


Kinase-catalyzed Biotinylation with Inactivated Lysates for Discovery of Substrates (K-BILDS).
Protein phosphorylation is catalyzed by kinases to regulate a large variety of cellular activities, including growth and signal transduction. Methods to identify kinase substrates are crucial to fully understand phosphorylation-mediated cellular events and disease states. Here, we report a set of protocols to identify substrates of a target kinase using Kinase-catalyzed Biotinylation with Inactivated Lysates for Discovery of Substrates (K-BILDS). As described in these protocols, K-BILDS involves inactivation of endogenous kinases in lysates, followed by addition of an active exogenous kinase and the γ-phosphate-modified ATP analog ATP-biotin for kinase-catalyzed biotinylation of cellular substrates. Avidin enrichment isolates biotinylated substrates of the active kinase, which can be monitored by western blot. Substrates of the target kinase can also be discovered using mass spectrometry analysis. Key advantages of K-BILDS include compatibility with any lysate, tissue homogenate, or complex mixture of biological relevance and any active kinase of interest. K-BILDS is a versatile method for studying or discovering substrates of a kinase of interest to characterize biological pathways thoroughly. © 2023 Wiley Periodicals LLC. Basic Protocol 1: FSBA treatment of lysates to inactivate kinases Basic Protocol 2: Kinase-catalyzed Biotinylation with Inactivated Lysates for Discovery of Substrates (K-BILDS).


Stem cells and veterinary medicine: tools to understand diseases and enable tissue regeneration and drug discovery.
New sophisticated laboratory techniques, as well as established interactions between basic science, researchers and veterinarians, have led to an exponential increase in our understanding of the animal body in health and disease. The advent of animal cloning, the identification and characterization of stem cells, and publication of the various mammalian genomes has afforded the opportunity to exploit these technologies to better understand disease and develop new therapies. In human medicine, these medical advances are already being translated into clinical practice, the promise being that previously untreatable or incurable chronic diseases will become a thing of the past. In parallel, the veterinary profession is looking to these technologies to explore novel therapies for chronic diseases, such as osteoarthritis in companion animals, and is applying these technologies to enhance food animal production. This review focuses on the emerging area of stem cell biology and explores the potential applications of stem cell technologies to veterinary medicine.


Increased in vivo secretory activity of neutrophil granulocytes in patients with psoriasis and palmoplantar pustulosis.
The relationship between psoriasis and palmoplantar pustulosis (PPP) is uncertain, as is the role of the neutrophil granulocyte in these conditions. In a previous comparative study of the rate of polymorphonuclear leucocyte (PMN) phagocytosis of IgG- and IgG-C3b-coated particles, an increased uptake rate was found in both diseases. Further information on the in vivo activity of PMNs in these conditions may be obtainable by determining the level of lactoferrin (LF) in serum from such patients, since LF serves as a specific marker of the turnover and activity of the circulating pool of neutrophils. In this study on 19 patients with psoriasis and 20 patients with PPP, elevated levels of LF were found in both conditions. In contrast, the levels of lysozyme and beta 2-microglobulin, which are markers of monocyte-macrophage and lymphocyte activity, respectively, were normal. This suggests the selective activation of neutrophils in these disorders. LF was significantly correlated (P less than 0.05 and 0.001, respectively) to the rates of phagocytosis of IgG- and IgG-C3b-coated particles, but not to the chemotaxis of isolated PMNs. There was no correlation between the severity of the disease and the levels of serum LF. The data suggest the increased in vivo activity of neutrophils in psoriasis and PPP.


Contributions to arachidonic acid release in mouse cerebrum by the phosphoinositide-phospholipase C and phospholipase A2 pathways.
Recent studies have indicated two major mechanisms for the release of arachidonic acid (20:4) from membrane phospholipids: 1) activation of phospholipase A2 and 2) stimulated hydrolysis of poly-phosphoinositides (PI) and diacylglycerols (DG) through phospholipase C and diacylglycerol lipase, respectively. In mammalian brain both mechanisms seem to be operable, although the relative contributions by these two pathways have not been carefully assessed. In this study three experimental protocols were used to examine 20:4 release in brain due to ischemia and agonist stimulation, as well as the metabolic relationship between this release and the increase in diacylglycerols, lysophospholipids, and inositol phosphates. The preferential release of arachidonic acid during the initial phase after decapitation was attributed mainly to the sequential hydrolysis of poly-PI to DG. During the second phase, the release of 20:4 along with other free fatty acids (FFA) correlated well with the increase in labeled lysophospholipids, suggesting the involvement of phospholipase A2. Diacylglycerols in brain are enriched in 18:0 and 20:4. Decapitation induced a rapid increase in the level of DG, which remained elevated during the 30 min period under study. Between 5 sec and 5 min, the increase in FFA lagged behind that of DG. The parallel increases in 18:0 and 20:4 in the FFA pool further support the notion that, during the early phase, 20:4 could be derived from the sequential hydrolysis of poly-PI and DG. Decapitation also induced a sequential appearance of Ins(1,4,5)P3, Ins(1,4)P2, and Ins(4)P, which peaked at 30 sec, 1 min, and 2 min, respectively. The level of 20:4 in brain was also examined with respect to poly-PI turnover due to stimulation by cholinergic agonists. Administration of pilocarpine to lithium-treated mice resulted in increased accumulation of labeled inositol monophosphate (IP1) compared to the amount in controls receiving lithium alone, as well as a less obvious increase in 20:4. Both pilocarpine-mediated increases (IP1 and 20:4) could be blocked by atropine. These results point to the presence of an active mechanism for poly-PI turnover and for the recycling of 20:4 in brain.


A randomised, double blind, placebo controlled trial with vitamin D3 as an add on treatment to interferon β-1b in patients with multiple sclerosis.
To study the safety and efficacy of vitamin D3 as an add on therapy to interferon β-1b (IFNB) in patients with multiple sclerosis (MS). 1 year, double blind, placebo controlled, randomised study in 66 MS patients. The primary outcomes were T2 burden of disease (BOD) on MRI scans, proportion of patients with serum levels of 25-hydroxyvitamin D (25(OH)D) ≥85 nmol/l or intact parathyroid hormone (PTH) ≤20 ng/l, and number of adverse events. Secondary outcomes were number of MRI enhancing T1 lesions and new T2 lesions, annual relapse rate, changes in the Expanded Disability Status Scale score, timed 25 foot walk test and timed 10 foot tandem walk tests. Median change in BOD was 287 mm(3) in the placebo group and 83 mm(3) in the vitamin D group (p=0.105). Serum levels of 25(OH)D increased from a mean of 54 (range 19-82) nmol/l to 110 (range 67-163) nmol/l in the vitamin D group. 84% of patients reached a serum 25(OH)D level >85 nmol/l in the vitamin D group and 3% in the placebo group (p<0.0001). Patients in the vitamin D group showed fewer new T2 lesions (p=0.286) and a significantly lower number of T1 enhancing lesions (p=0.004), as well as a tendency to reduced disability accumulation (p=0.071) and to improved timed tandem walk (p=0.076). There were no significant differences in adverse events or in the annual relapse rate. Vitamin D3 add on treatment to IFNB reduces MRI disease activity in MS. EudraCT number 2007-001958-99 and ClinicalTrialsGov number NCT01339676.


Correction to: Alpha-1 Antitrypsin Deficiency: a Rare Disease?
The original version of this article contained errors in Table 3 on page 6.


Anatomic effect of distention therapy in unstable bladder: new approach.
The recent clinical success of distention therapy in the treatment of the unstable bladder is reviewed. Bladder stability and increased capacity as measured by cystometry following distention therapy as well as relief of symptoms have prompted this anatomic study. The neuromuscular pathways of conduction in rat and rabbit bladder wall were examined following short-term (two-hour) and prolonged (six-hour) distention. Treated and control animals were studied at fixed intervals for four months. Prolonged distention did not alter either smooth muscle cell architecture or intercellular junctions. It did produce a transient phase of degeneration among the unmyelinated nerve fibers in the bladder wall consisting of axonal swelling and lysis of organelles. A quantitative estimate of nerve injury was compiled using pooled histograms. These results suggest that bladder stability following distention therapy may be related to nerve degeneration in the bladder wall.


Pantothenate biosynthesis in higher plants.
Pantothenate (vitamin B5) is a water-soluble vitamin essential for the synthesis of CoA and ACP (acyl-carrier protein, cofactors in energy yielding reactions including carbohydrate metabolism and fatty acid synthesis. Pantothenate is synthesized de novo by plants and micro-organisms; however, animals obtain the vitamin through their diet. Utilizing our knowledge of the pathway in Escherichia coli, we have discovered and cloned genes encoding the first and last enzymes of the pathway from Arabidopsis, panB1, panB2 and panC. It is unlikely that there is a homologue of the E. coli panD gene, therefore plants must make beta-alanine by an alternative route. Possible candidates for the remaining gene, panE, are being investigated. GFP (green fluorescent protein) fusions of the three identified plant enzymes have been generated and the subcellular localization of the enzymes studied. Work is now being performed to elucidate expression patterns of the transcripts and characterize the proteins encoded by these genes.


Changes in subcellular accumulation of contractile proteins in myocardiocyte cultures: effects of fibric acid derivatives.
We analyzed the influence of 6 and 24 h of treatment with the fibric acid derivatives bezafibrate (10 micrograms/ml), gemfibrozil (23 micrograms/ml), and fenofibrate (30 micrograms/ml) on alpha-actinin, troponin-T, and tropomyosin proteins in the cytoplasmic and cytoskeletal fractions of cultured chick myocardiocytes. The findings with sodium dodecyl sulfate-gel electrophoresis and immunoblotting showed that all three drugs modified cellular and subcellular protein levels in different ways: bezafibrate and fenofibrate produced the most significant alterations in both fractions, modifying alpha-actinin, troponin T, and tropomyosin compartmentalization in myocardiocytes, whereas gemfibrozil altered these proteins less notably. Given the role of these proteins in heart muscle contraction, fibric acid derivative-induced changes may be related with the secondary effects of these drugs on heart rhythmicity.


A novel approach to improve hand hygiene compliance of student nurses.
The National University Hospital, Singapore routinely undertakes standardized Hand Hygiene auditing with results produced by ward and by staff type. In 2010 concern was raised over consistently low compliance by nursing students averaging 45% (95% CI 42%-48%) prompting us to explore novel approaches to educating our next generation of nurses to improve their hand hygiene practice.We introduced an experiential learning assignment to final year student nurses on attachment to NUH inclusive of hand hygiene auditor training followed by a period of hand hygiene observation. The training was based on the World Health Organisation (WHO) "My 5 moments for hand hygiene" approach. Upon completion students completed an anonymous questionnaire to evaluate their learning experience. By 2012, nursing students were 40% (RR: 1.4, 95% CI 1.3-1.5, p<0.001) more likely to comply with hand hygiene practices. 97.5% (359/368) of nursing students felt that the experience would enhance their own hand hygiene practice and would recommend participating in audits as a learning instrument. With consideration of all stakeholders a sustainable, flexible, programme was implemented. Experiential learning of hand hygiene was a highly valued educational tool and in our project was directly associated with improved hand hygiene compliance. Feedback demonstrated popularity amongst participants and success in achieving its program objectives. While this does not guarantee long term behavioural change it is intuitive that instilling good habits and messages at the early stages of a career will potentially have significant long-term impact.


SARS-CoV-2-Naïve Korean Children and Adolescents Hospitalized With COVID-19 in 2021.
The risk of severe outcomes with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) delta variant remains low in children and adolescents, but less is known about its effect on the SARS-CoV-2-naïve population. This study evaluated clinical manifestations and risk factors for moderate-to-critical coronavirus disease 2019 (COVID-19) in mostly SARS-CoV-2-naïve children and adolescents in 2021. This multicenter retrospective study included patients aged 0-18 years who were hospitalized with COVID-19 at 8 referring hospitals in South Korea during the predelta-predominant and delta-predominant periods in 2021. Each case was labeled as either hospitalization with medical needs or for isolation. Severity was categorized as mild, moderate, severe, or critical with regard to pneumonia presence and illness severity. Among 753 cases, most (99.5%) had no prior history of COVID-19 or vaccination against COVID-19. The proportions of hospitalization with medical needs (3.5% vs. 19.7%), moderate illness (0.9% vs. 4.0%), and severe/critical illness (0.8% vs. 5.3%) increased during delta predominance. The risk of moderate-to-critical COVID-19 among hospitalizations with medical needs was higher among patients aged 12-18 years (adjusted odds ratio [aOR], 4.1; 95% confidence interval [CI], 1.5-11.8) and with obesity (aOR, 6.9; 95% CI, 2.4-19.6) but not among patients infected during delta predominance. However, children with obesity experienced more severe COVID-19 during delta predominance (aOR, 6.1; 95% CI, 1.2-29.6). Despite its similar severity among most SARS-CoV-2-naïve children and adolescents, the delta variant may affect COVID-19 severity in those with high-risk underlying medical conditions. Underlying conditions, particularly obesity, may cause severe COVID-19 in children and adolescents, warranting strong consideration for vaccinating high-risk children.


Leaching and degradation of S-Metolachlor in undisturbed soil cores amended with organic wastes.
Organic waste (OW) reuse in agriculture is a common practice fostered by benefits in terms of waste recycling and crop production. However, OW amendments potentially affect the fate of pesticide spread on fields to protect the crops from pests and weeds. The influence of OW on the sorption, degradation, and leaching of pesticides is generally studied for each mechanism separately under artificial laboratory conditions. Our study aims at evaluating the balance of these mechanisms under more realistic conditions to clarify the influence of three common OW amendments on the fate, in soil, of the widely used herbicide S-Metolachlor. We performed leaching experiments in large undisturbed soil cores amended with raw sewage sludge, composted sludge, and digested pig slurry (digestate), respectively. We monitored S-Metolachlor and its two main metabolites MET-OA and MET-ESA in the leachates during a succession of 10 rainfall events over 126 days. We also quantified the remaining S-Metolachlor and metabolites in the soil at the end of the experiments. S-Metolachlor leaching didn't exceed 0.1% of the applied dose with or without OW amendment. Despite a soil organic carbon increase of 3 to 32%, OW amendments did not significantly affect the amount of S-Metolachlor that leached through the soil (0.01 to 0.1%) nor its transformation rate (6.0 to 8.6%). However, it affected the degradation pathways with an increase of MET-OA relative to MET-ESA formed after OW amendment (28 to 54%) compared to the controls (8%). Concentration of S-Metolachlor and metabolites in the leachates of all treatments greatly exceeded the regulatory limit for groundwater intended for human consumption in Europe. These high concentrations were probably the consequence of preferential macropore flow. Colloids had comparable levels in the leachates after S-Metolachlor application. Dissolved organic carbon was also comparable in the controls, digestate, and sludge treatments but was 65% higher in the compost-amended cores. These results, along with a great variability among replicates inherent to experiments performed under realistic conditions, partly explain the limited impact of OW on the transport of S-Metolachlor.


Elective Cardioversion in the Era of Novel Oral Anticoagulants - Does a Nurse Administered Verbal Questionnaire for Compliance Negate the Need for Routine Transoesophageal Echocardiography?
Anticoagulation prior to elective external direct current cardioversion (EDCCV) is mandatory. The inability to monitor compliance with novel oral anticoagulants (NOACs) raises a potential safety issue. We aimed to evaluate whether a structured, nurse-led assessment of compliance prior to EDCCV ensures safety without the need for routine transoesophageal echocardiography (TOE). Data was prospectively collected on consecutive patients undergoing EDCCV during 2014-2015. All procedures were supervised by an electrophysiology clinical nurse consultant (EPCNC). Drug compliance was verbally assessed using a standardised questionnaire by the EPCNC. Novel oral anticoagulants compliance was required for a continuous period of 3 weeks prior to EDCCV; otherwise a TOE-guided EDDCV was performed. All patients had follow-up 30 days post-procedure. Three hundred and eleven cardioversions were performed on 256 patients in whom 154 (49.5%) were prescribed a NOAC (rivaroxaban (n=105; 68.2%), dabigatran (n=38; 24.7%), apixaban (n=11; 7.1%)). Median age was 63 years (24-94 yrs), mean CHADS2-Vasc score was 2.0±1.5 and 138 (89.6%) were outpatients. One hundred and twenty-nine (83.8%) EDCCV were for atrial fibrillation and 25 (16.2%) for atrial flutter. Sinus rhythm was achieved in 90.3% of cases. Fourteen patients (9%) assessed as non-compliant underwent TOE. 129 (83.8%) EDCCV were performed without prior TOE. No stroke or systemic embolism was identified in any patient treated with either warfarin or a NOAC. A standardised, verbal questionnaire can be administered to detect NOAC non-compliance in patients undergoing EDCCV. With appropriate compliance assessment a nurse-led EDCCV without routine TOE did not significantly compromise safety in this study group.


Lipomatous ganglioneuroma of the retroperitoneum.
Lipomatous ganglioneuroma (LG) is a rare variant of ganglioneuroma that is histologically characterized by a mature adipocytic component admixed with a conventional ganglioneuroma. We report the clinicopathological and immunohistochemical features of an LG in a 44-year-old Chinese male; additionally, we review the literature regarding this type of tumor. Magnetic resonance imaging revealed a left paravertebral soft-tissue mass at the T11-L3 levels. Grossly, the encapsulated neoplasm had a white to yellowish cut surface and rubbery consistency. Microscopic evaluation revealed an encapsulated lesion that consisted of areas of ganglioneuroma admixed with areas of mature fat. By immunohistochemistry, the ganglion cells were positive for chromogranin and synaptophysin, whereas the Schwann cells were positive for vimentin, S-100 protein, and glial fibrillary acidic protein (GFAP). This is the second known report of a retroperitoneal LG. The patient was well and without evidence of disease at 2 years' follow-up.


Discovery of novel coffee diterpenoids with inhibitions on Cav3.1 low voltage-gated Ca2+ channel.
Seven new (1-4, 6-8) diterpenoids with rare skeletons and seven known ones (9, 12, 17, 18 and 23-25) were isolated from roasted beans of Coffea arabica L. Together with previously obtained diterpenoids, a total of 26 molecules (1-25, 4a) were evaluated their activities on Cav3.1 low voltage-gated Ca2+ channel. Compounds 1, 3, 6, 7, 12, 13, 17, 19 and 24 exhibited noticeable Cav3.1 inhibitions (41.2%-96.1%) at 10 μM. The IC50 values of 1, 6, 7, 12, 13, 17 and 24 are 2.9, 2.3, 0.68, 14.8, 11.6, 6.1 and 6.8 μM, respectively. The ring moiety at C-18 and C-19, and esterification of OH-17 with long-chain fatty acids seem important for their activities. Further studies indicated that 1 and cafestol may act on different binding sits with the Cav3.1 blocker Z944, which is in clinical trial. Significantly, the present study initially shows that coffee diterpenoids are potential natural resources for Cav3.1 inhibitors.


Long-term outcome after surgical closure of atrial septal defect in childhood with extensive assessment including MRI measurement of the ventricles.
Surgical closure of the secundum type of atrial septal defect (ASD) in childhood leads to excellent survival. However, relevant morbidity has been reported. Transcatheter closure of these defects has now become an alternative approach. To compare the results of the two different interventions, reliable data are needed on the long-term morbidity after defect closure with both methods. Patients were evaluated after a minimum of 10 years after surgical closure of an ASD in childhood. Assessment included analysis of perioperative data, interview, clinical examination, electrocardiogram, (ECG), 24-hour ECG, ergometry, chest radiograph, echocardiography, and MRI. A total of 66 patients underwent operation between 1971 and 1986 at our institution. Forty-eight of them (73%) were interviewed and 38 (58%) participated fully in the study program. Eighteen (27%) either refused to participate or were lost to follow-up. There were no substantial residual disorders, such as arrhythmias, right-sided heart dilatation, pulmonary hypertension, or reduced work capacity. Surgical closure of an ASD in childhood has an excellent long-term outcome. Surgical closure is thus the standard against which transcatheter closure needs to be measured.


Female hormone release of microencapsulated Xenopus laevis ovarian cells.
Cell microencapsulation has potential for the treatment of endocrine diseases. This study aims to probe the feasibility of Xenopus laevis as an animal model for cell microencapsulation and transplantation and to evaluate the female hormone release of microencapsulated X. laevis ovarian cells. The cells were harvested, cultured and microencapsulated into alginate-chitosan-alginate microcapsules with an electrostatic generator. The estradiol and progesterone releases of the microencapsulated X. laevis ovarian cells were investigated both in vitro and in vivo. The results showed that the microencapsulated cells kept secreting estradiol and progesterone in vitro for 60 days. After transplantation, serum estradiol and progesterone levels in ovariectomized X. laevis remained elevated for 60 days. X. laevis has been proved to be a suitable animal model for cell microencapsulation and transplantation. Microencapsulated ovarian cells may be considered as a promising endogenous drug delivery system for the treatment of deficiency of female hormones.


Prognostic Value of Hyperreflective Material on Visual Acuity in Treatment-Naïve BRVO.
To correlate between hyperreflective material (HRM) seen on optical coherence tomography (OCT) and visual outcomes in treatment-naïve acute branch retinal vein occlusion (BRVO). Demographic and OCT features of patients with acute BRVO were analyzed retrospectively. OCT parameters noted were macular edema type, HRM presence and location, and outer retinal layer integrity. Eyes were separated into two groups: Group 1 (BRVO without HRM) and Group 2 (BRVO with HRM). Forty-seven eyes (67%) were included in Group 1 and 23 eyes (33%) were included in Group 2. Mean presenting logMAR visual acuity in Groups 1 and 2 was 0.521 ± 0.428 (mean Snellen equivalent = 20/66) and 0.627 ± 0.427 (mean Snellen equivalent = 20/85), respectively. Poor vision at final visit was noted in the BRVO with HRM group (P = .027). More intravitreal injections were required for macular edema resolution in the HRM group (4.83 vs. 2.67). Presence of HRM in BRVO is associated with poorer vision. It can be a useful prognostic biomarker. [Ophthalmic Surg Lasers Imaging Retina. 2020;51:320-327.].


A single-center retrospective comparative analysis of urinary continence in robotic prostatectomy with a combination of umbilical ligament preservation and Hood technique.
Data available on the effect of the recently developed Hood technique and its modified iterations in robot-assisted radical prostatectomy on postoperative urinary continence are insufficient. We evaluated the time to achieve urinary continence with the modified Hood technique compared with the standard or umbilical ligament preservation robot-assisted radical prostatectomy. This retrospective analysis examines patient records for those who underwent robot-assisted radical prostatectomy at the Jyoban Hospital of Tokiwa Foundation in Fukushima, Japan, from 2017 to 2021. The main outcome was to determine significant differences in the time taken to achieve urinary continence among the three procedure types. We employed the Kaplan-Meier survival analysis to estimate the time to achieve urinary continence in the three procedure types of robot-assisted radical prostatectomy. Additionally, we used a Cox regression hazard model to evaluate the association between the time to achieve urinary continence and the procedure types. We considered 196 patients in this study. The estimated rates of achieving urinary continence at 6 months following standard, umbilical ligament preservation, and modified Hood technique robot-assisted radical prostatectomy were 77.6%, 89.5%, and 100%, respectively. The multivariable Cox hazard regression model showed that patients who underwent the modified Hood technique were significantly more likely to achieve urinary continence than those who underwent the standard robot-assisted radical prostatectomy. The modified Hood technique achieved better urinary continence outcomes, with all patients with the procedure achieving urinary continence at 6 months. Further randomized controlled trials are required to validate this finding.


Epithelial transdifferentiation and cataract in the human lens.
Anterior subcapsular cataracts cause a serious loss of vision and are normally associated with ocular trauma, inflammation or clinical skin conditions. They appear to be accompanied by epithelial cell growth and transdifferentiation where unscheduled production of a number of proteins, including alpha smooth muscle actin (alpha-sma), occurs. Clinical studies have also revealed an up-regulation of the TGFbeta signalling pathway in such cataracts. The present study, using phase contrast and immunofluorescent techniques, was undertaken to investigate the extent of alpha-sma expression in traumatic cataracts, in capsulorhexis specimens obtained during cataract surgery and in aged human lenses from donor eyes. The donor lenses were also exposed to trauma or TGFbeta in culture to observe their relative contribution to alpha-sma production. Dense anterior subcapsular cataracts were relatively rare (<1%), but all showed a pronounced up-regulation of alpha-sma, which was located both in anterior cells of normal appearance and in nucleated fibroblastic cells lying beneath the anterior epithelium. Surprisingly, more than 50% of capsulorhexis specimens from mature cataracts showed expression of alpha-sma, although to a limited extent. Alpha-sma was not expressed in any of the clear donor lenses and culture for 8 days in EMEM did not induce expression. Interestingly, unlike their young animal counterparts, human lenses failed to show the presence of alpha-sma when exposed to 10 ng ml(-1) TGFbeta. However, after culture, lenses with pre-existing cortical opacities did express alpha-sma, as did clear lenses subjected to injury or trauma. It appears that the greater the stress, the greater is the expression of alpha-sma. Cataract, and especially cortical cataract, should therefore be seen as associated with stress-induced signalling pathways in the lens that lead to the transdifferentiation of the anterior epithelial cells.


Breath stacking in children with neuromuscular disorders.
Respiratory muscle weakness in neuromuscular disorders (NMD) can lead to shallow breathing and respiratory insufficiency over time. Children with NMD often cannot perform maneuvers to recruit lung volume. In adults, breath stacking with a mask and one-way valve can achieve significantly increased lung volumes. To evaluate involuntary breath stacking (IBS) in NMD, we studied 23 children of whom 15 were cognitively aware and able to communicate verbally. For IBS, a one-way valve and pneumotachograph were attached to a face mask. Tidal volumes (Vt) and minute ventilation (VE ) were calculated from airflow over 30 sec before and after 15 sec of expiratory valve closure. Six cooperative male subjects with Duchenne muscular dystrophy (DMD) participated in a subsequent comparison of IBS with voluntary breath stacking (VBS) and supported breath stacking (SBS). The average Vt in those studied with IBS was 277 ml (range 29-598 ml). The average increase in volume by stacking was 599 ml (range -140 to 2,916 ml) above Vt . The average number of stacked breaths was 4.5 (range 0-17). VE increased on average by 18% after stacking (P < 0.05, paired t-test). Oxygen saturation did not change after stacking. Four of the 23 children did not breath stack. Compared to IBS, VBS achieved similar volumes in the six subjects with DMD but SBS was more successful in those with greatest muscle weakness. IBS may achieve breath volumes of approximately three times Vt and may be particularly useful in non-cooperative subjects with milder degrees of respiratory muscle weakness.


A modification of the amidoblack-TCA-staining for quantitative microspectrometrical determination of proteins in tissue sections.
Conditions are described by which cells and fresh frozen tissues, following fixation with ethanol-ether, and after staining with the amidoblack (AB)-TCA-staining method, show a modified dye/protein ratio of 0.9 moles AB/10(5) g protein compared to 8.5 moles AB/10(5) g protein ( Schauenstein et al. 1980) as in a previously used method. In contrast to the AB-TCA-method, which leads to extremly high and unmeasurable extinctions in tissue sections, staining with the modified AB-TCA- 23st -method with 10 microns tissue sections produces easily measurable extinction values. A correlation of the microspectrometrically determined mean total extinction values of different cell types and nuclei after staining with the tetrazonium method (N ohammer 1978; N ohammer and Desoye 1981) and on the other hand with the AB-TCA 23st -method has been found. The microspectrometrically determined extinctions after AB-TCA 23st -staining can be calculated; an extinction of 0.04248 corresponds to 1 pgm protein.


Left-sided intraventricular conduction delay concealed by right bundle branch block.
The electrocardiograms of a 69-year-old woman with severe heart failure showed a left-sided nonspecific intraventricular conduction delay (QRS duration, 0.13 seconds) characterized by notches and slurrings in the descending part of the R waves in the left precordial leads and in the ascending parts of the S waves in the right precordial leads. Subsequently, a complete right bundle branch block with wider QRS complexes (0.17 seconds) appeared. It is possible to consider that the left-sided nonspecific intraventricular conduction delay was concealed by the greater conduction delay occurring when the complete right bundle branch block was present. This would be in keeping with a previous study using Doppler tissue imaging in which this was postulated but where the corresponding electrocardiograms were not shown.


Phycicoccus mangrovi sp. nov., a novel endophytic actinobacterium isolated from bark of Sonneratia apetala.
Two novel strains KQZ13P-1T and MAQZ13P-2 were isolated from bark of Sonneratia apetala collected from Maowei sea Mangrove Nature Reserve in Guangxi Zhuang Autonomous Region, China. Two strains were Gram-positive, aerobic, non-spore-forming, no diffusion pigment actinobacterial strains and investigated by a polyphasic approach to determine their taxonomic position. The average nucleotide identity (ANI) value and the digital DNA-DNA hybridization (dDDH) value between the two strains were 99.9% and 99.7%, respectively, suggesting that they belonged to the same species. The ANI and dDDH values between strain KQZ13P-1T and five Phycicoccus species were 74.4-95.3% and 20.1-61.5%, respectively. Phylogenetic analyses based on 16S rRNA gene sequences showed that the two strains were member of the genus Phycicoccus and were closely related to P. jejuensis NRRL B-24460T (99.2% sequence similarity), followed by P. ginsengisoli DCY87T (97.5-97.6%). Moreover, based on 88 core genes, the phylogenomic tree indicated that the two strains clustered with P. jejuensis NRRL B-24460T. The cell-wall peptidoglycan of both strains contained meso-diaminopimelic acid. The major fatty acids in two strains were C17:1ω8c, iso-C15:0 and iso-C16:0. The major polar lipids included diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE) and phosphatidylinositol (PI). Based on phylogenetic, phenotypic and chemotaxonomic analysis, strains KQZ13P-1T and MAQZ13P-2 represent a novel species of the genus Phycicoccus, for which the name Phycicoccus mangrovi sp. nov. is proposed. The type strain is KQZ13P-1T (=CGMCC 1.18973T = JCM 34556T).


Characterization of agitation environments in 250 ml spinner vessel, 3 L, and 20 L reactor vessels used for animal cell microcarrier culture.
Three dimensional particle tracking velocimetry (3-D PTV) was used to characterize the flow fields in the impeller region of three microcarrier reactor vessels. Three typical cell culture bioreactors were chosen: 250 ml small-scale spinner vessels, 3 L bench-scale reactor, and 20 L medium-scale reactor. Conditions studied correspond to the actual operating conditions in industrial setting and were determined based on the current scale-up paradigm: the Kolmogorov eddy length criterion. In this paper we present characterization of hydrodynamics on the basis of flow structures produced because of agitation. Flow structures were determined from 3-D mean velocity results obtained using 3-D PTV. Although the impellers used in 3 L and 20 L reactors were almost identical, the flow structures produced in the two reactors differed considerably. Results indicate that near geometric scale up does not necessarily amount to scale-up of flow patterns and indicates that intensity as well as distribution of energy may vary considerably during such a scale-up.


Healthcare use and clinical investigations before a diagnosis of ovarian cancer: a register-based study in Denmark.
Ovarian cancer (OC) is associated with a poor prognosis, which calls for earlier diagnosis. This study aimed to analyse the health care use in primary care and at hospitals among women with OC compared to non-cancerous women to identify a window of opportunity for earlier diagnosis. This nationwide register-based observational cohort study included all Danish women aged ≥ 40 years who were diagnosed with a first-time OC or borderline ovarian tumour in 2012-2018 and with no previous cancer diagnosis (n = 4,255). For each case, ten non-cancerous women were identified (n = 42,550). We estimated monthly incidence rate ratios using a negative binomial regression model to assess the use of health care services. We calculated risk ratios of having multiple contacts to general practice before a diagnosis using a binary regression model. Cases had statistically significantly higher contact rates to general practice from five months prior to the diagnosis compared to references. From six to eight months prior to diagnosis, an increased use of transvaginal ultrasound and gynaecologist was seen for cases compared to references. Increased healthcare use was seen relatively closely to the time of diagnosis for women with OC. This indicates a narrow window of opportunity for a timelier diagnosis. Still, the use of specialised assessment increased at six to eight months before the diagnosis. When women present unspecific symptoms, awareness of potential ovarian malignancies and safety-netting by the general practitioner may be pivotal. Not relevant.


An endogenous membrane-bound protein inhibits the phosphorylation of the L-type calcium channel by the cAMP-dependent protein kinase.
An endogenous protein inhibits the PKA-phosphorylation of the DHP-binding calcium channel complex in vitro. The inhibitory activity could be reduced by a treatment with detergents or dithiothreitol. Further purification separates the inhibitory activity from the dihydropyridine-binding calcium channel complex. Both activities are localized in the plasma membrane indicating that this protein kinase-inhibitor could interfere with the phosphorylation of the calcium channel by the cAMP-dependent protein kinase. The inhibitory activity may therefore take part in the regulation of the calcium channel current.


Dopaminergic impact on local and global cortical circuit processing during learning.
We have learned to detect, predict and behaviorally respond to important changes in our environment on short and longer time scales. Therefore, brains of humans and higher animals build upon a perceptual and semantic salience stored in their memories mainly generated by associative reinforcement learning. Functionally, the brain needs to extract and amplify a small number of features of sensory input with behavioral relevance to a particular situation in order to guide behavior. In this review, I argue that dopamine action, particularly in sensory cortex, orchestrates layer-dependent local and long-range cortical circuits integrating sensory associated bottom-up and semantically relevant top-down information, respectively. Available evidence reveals that dopamine thereby controls both the selection of perceptually or semantically salient signals as well as feedback processing from higher-order areas in the brain. Sensory cortical dopamine thereby governs the integration of selected sensory information within a behavioral context. This review proposes that dopamine enfolds this function by temporally distinct actions on particular layer-dependent local and global cortical circuits underlying the integration of sensory, and non-sensory cognitive and behavioral variables.


Knowledge-based approach to septic shock patient data using a neural network with trapezoidal activation functions.
In this contribution we present an application of a knowledge-based neural network technique in the domain of medical research. We consider the crucial problem of intensive care patients developing a septic shock during their stay at the intensive care unit. Septic shock is of prime importance in intensive care medicine due to its high mortality rate. Our analysis of the patient data is embedded in a medical data analysis cycle, including preprocessing, classification, rule generation and interpretation. For classification and rule generation we chose an improved architecture based on a growing trapezoidal basis function network for our metric variables. Our results extend those of a black box classification and give a deeper insight in our patient data. We evaluate our results with classification and rule performance measures. For feature selection we introduce a new importance measure.


Fission track estimation of uranium concentrations in drinking water from Azad Kashmir, Pakistan.
The analysis of uranium in water samples can be very helpful for providing guidelines to the general public regarding necessary remedial measures. A fission-track technique has been applied for the estimation of the uranium concentration in drinking water collected from natural springs of Muzaffarabad and hilly areas of Reshian, Azad Kashmir. The technique involved simultaneous irradiation with thermal neutrons of a sample and a standard in contact with a track detector, and the counting of the fission tracks in the detector from the (n, f) nuclear reaction. Uranium concentrations of the samples were determined by comparing fission-track density with that of a standard of known uranium concentration. Uranium concentration in water samples from the Muzaffarabad and Reshian area varied from 0.03 +/- 0.01 microgL(-1) to 6.67 +/- 0.14 microgL(-1) with an average of 1.36 +/- 0.05 microgL(-1). The observed concentrations of uranium in drinking water were found to be less than the Maximum Acceptable Concentration levels of 9-30 microgL(-1). Thus, the observed values are within safe limits as far as uranium related health hazards are concerned.


Construction and validation of an aging-related gene signature for prognosis prediction of patients with breast cancer.
Breast cancer (BC) is an aging-related disease. Aging-related genes (ARGs) participate in the initiation and development of lung and colon cancer, but the prognosis signature of ARGs in BC has not been clearly studied. This study aimed to construct an ARGs signature to predict the prognosis of patients with breast cancer. Firstly, the expression data of ARGs from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) were collected. Then COX and least absolulute shrinkage and selection operator(LASSO) were performed to construct the ARGs prognostic signature. The correlation between the signature and immune cell infiltration, immunotherapeutic response and drug sensitivity were subsequently analysed. The TCGA nomogram was constructed by combining the signature with other clinical features, and was validated by using GEO database. After LASSO and COX regression analyses, a prognostic signature based on nine ARGs, namely, HSP90AA1, NFKB2, PLAU, PTK2, RECQL4, CLU, JAK2, MAP3K5, and S100B, was built by using the TCGA dataset. Moreover, this risk signature is closely related to immune cell infiltration, immunotherapeutic response, and responses to chemotherapy and targeted therapy. Subsequently, The calibration curve demonstrates that the nomogram agrees well with practical prediction results. The receiver operating characteristic curve and decision-making curve analysis demonstrate that ARG signature has the better prognosis diagnosis ability and clinical net benefits. Therefore, the proposed ARG prognosis signature is a new prognosis molecular marker of patients with BC, and it can provide good references to individual clinical therapy.


Correction: Functional Analysis of Free Fatty Acid Receptor GPR120 in Human Eosinophils: Implications in Metabolic Homeostasis.
[This corrects the article DOI: 10.1371/journal.pone.0120386.].


Rapamycin analogs with reduced systemic exposure.
The synthesis and biological activities of rapamycin (I) analogs modified at the C-40 position are reported. Emphasis placed on compounds that potentially have an improved safety profile on account of their shorter in vivo half-life when compared with rapamycin.


The down-regulation of casein kinase 1 alpha as a host defense response against infectious bursal disease virus infection.
Infectious bursal disease virus (IBDV) is an important immunosuppressive virus of chickens. Although the gene functions of IBDV have been well characterized, the host responses during IBDV infection remain much poor. In the present study, casein kinase 1 alpha (CK1α), a novel VP2-associated protein, was down-regulated during IBDV replication in DF1 cells. Further experiments showed that siRNA-mediated knockdown of CK1α inhibited IBDV replication, while overexpression of CK1α promoted IBDV growth. Finally, we revealed that the effects of CK1α expression level on IBDV replication were involved in the negative regulation of CK1α on type I interferon receptor (IFNAR1), because ubiquitination assay analyses demonstrated that CK1α could promote the ubiquitination of IFNAR1, thereby affecting the stability of this receptor. In conclusion, down-regulation of CK1α during IBDV infection as a host defense response increased abundance of IFNAR1, which in turn enhanced an inhibitory effect on IBDV replication.


An IL-2 mutein engineered to promote expansion of regulatory T cells arrests ongoing autoimmunity in mice.
Interleukin-2 (IL-2) controls the homeostasis and function of regulatory T (Treg) cells, and defects in the IL-2 pathway contribute to multiple autoimmune diseases. Although recombinant IL-2 therapy has been efficacious in certain inflammatory conditions, the capacity for IL-2 to also activate inflammatory effector responses highlights the need for IL-2-based therapeutics with improved Treg cell specificity. From a panel of rationally designed murine IL-2 variants, we identified IL-2 muteins with reduced potency and enhanced Treg cell selectivity due to increased dependence on the IL-2 receptor component CD25. As an Fc-fused homodimer, the optimal Fc.IL-2 mutein induced selective Treg cell enrichment and reduced agonism of effector cells across a wide dose range. Furthermore, despite being a weaker agonist, overall Treg cell growth was greater and more sustained due to reduced receptor-mediated clearance of the Fc.IL-2 mutein compared with Fc-fused wild-type IL-2. Preferential Treg cell enrichment was also observed in the presence of activated pathogenic T cells in the pancreas of nonobese diabetic (NOD) mice, despite a loss of Treg cell selectivity in an IL-2R proximal response. These properties facilitated potent and extended resolution of NOD diabetes with infrequent dosing schedules.