Partnering of anammox and denitrifying bacteria benefits anammox's recovery from starvation and complete nitrogen removal.
The cooperative metabolic activity of anammox and denitrifying bacteria could speed up anammox's recovery and reduce nitrate generated from the anammox reaction. In this study, a laboratory-scale model system containing a defined anammox culture AMX and a simultaneous nitrification and denitrification (SND) bacterium - Thauera sp. strain SND5 was established and investigated. Several lines of evidence revealed that strain SND5 consumed soluble microbial products (SMPs) generated by culture AMX (as high as 1.5 mg/L), stimulating anammox activity after long-term starvation. At low C/N ratios with an optimal C/N of 1, SND5 completely consumed organic carbon first at anoxic condition, storing carbon intracellularly as poly-β-hydroxybutyrate (PHB) (as high as 0.6 mg/L biomass), thereby creating a favorable environment for the growth of anammox bacteria. The anammox reaction and nitrate reduction supported by PHB catabolism could then proceed simultaneously, resulting in enhanced nitrogen removal. Cooperative interactions between anammox and denitrifying bacteria involving SMPs consumption and PHB synthesis may play a significant role in nitrogen cycling at nitrite- and carbon-limited environments.


Inhibiting role of rosiglitazone in the regulation of inflammatory response and protective effects for severe acute pancreatitis in mice.
Our study aimed to probe the effects of rosiglitazone treatment on a severe acute pancreatitis (SAP) model induced by caerulein and investigate the underlying mechanism. Differentially expressed messenger RNAs (mRNAs) in the mice of a SAP group were screened out by microarray analysis. The inflammatory response pathway was obtained from the online website DAVID Bioinformatics Resources 6.8. The interactions of caerulein and its target proteins were shown by search tool for interactions of chemicals (STITCH). Functional interactions of the genes associated with pancreatitis and the target proteins of caerulein were obtained with search tool for interactions of chemicals (STRING). SAP mice were established by hourly intraperitoneal injection of caerulein. Rosiglitazone was used as treatment drug, and pancreatic inflammation was assessed. The expression of Socs3 was studied by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis. The expression of interleukin (IL)-6, IL-1b, and Egr1 were studied by RT-PCR and Western blot analysis. The GSE77983 data were analyzed, and the results showed that Socs3 was overexpressed in SAP tissues. The inflammation response pathway in pancreas was selected by DAVID, STITCH, and STRING. After injection of rosiglitazone in mice, the serum levels of amylase and lipase were decreased. Furthermore, the mRNA and protein levels of Socs3 and inflammatory cytokines in pancreatic tissues were downregulated. Rosiglitazone could protect mice with SAP from injury by downregulating Socs3 and inhibiting the inflammatory response pathway.


Gender and age disparity in health-related behaviors and behavioral patterns based on a National Survey of Taiwan.
This is a population-based study based on the 2002 National Survey of Taiwan on Knowledge, Attitude, and Practice of Health Promotion. The objective of this study is to examine health-related behaviors and behavioral patterns among different gender and age groups. A total of 26,755 participants were interviewed, resulting in a response rate of 81.9%. Factor analysis with orthogonal rotation was applied to identify the underlying factor structure for the health-related behaviors, including cigarette smoking, betel nut chewing, alcohol drinking, intake of fruits or vegetables, prevention service utilization, physical activity, and tooth brushing. Protective and risk behavioral patterns were selected consistently among various gender and age subgroups. These two behavior patterns were negatively associated with each other. In younger age groups (age < 55), a risk behavioral pattern was more dominant than a protective behavioral pattern. In the older age group (age >or= 55), the pattern order was reversed. An effective health intervention program should be based on behavioral patterns instead of an individual behavior. Gender and age play an important role in the behavioral patterns and need to be taken into consideration when designing intervention programs.


Partial characterization and identification of chemokinetic factors in serum.
The chemokinetic effect of serum, defined as the stimulation of the rate of polymorphonuclear neutrophil (PMN) migration in the presence of serum has been investigated for the purpose of identifying the major factors in serum contributing to this effect. The chemokinetic activity has been measured in fresh and heated (56 degrees C) serum and in fractions of serum separated by gel filtration. Gel filtration showed the existence of at least three heat-stable chemokinetic factors with approximate molecular weight of 800,000, 150,000, and 70,000. By immunoprecipitation and immunoadsorption the 800,000 mol. wt factor was identified as alpha 2-macroglobulin and the 70,000 mol wt. factor as alpha 1-antitrypsin and orosomucoid. Heat-labile chemokinetic activity was found in fractions representing mol. wts around 200,000 and 50,000. Immunoprecipitation and immunoadsorption indicated that these heat-labile chemokinetic factors were C3-associated. This investigation suggests a new functional role for the acute-phase reactants alpha 2-macroglobulin, alpha 1-antitrypsin, orosomucoid, and C3 in the modulation of PMN migration and also suggests that the chemokinetic activities of these substances are regulated by specific antagonists at various levels.


Non-classical MHC class I molecules on intestinal epithelial cells: mediators of mucosal crosstalk.
The mucosal immune environment consists of a complex combination of lymphoid cells, non-lymphoid cells, and lumenal bacteria. Signals from lumenal bacteria are constantly transmitted to the underlying tissues across the intestinal epithelial barrier. Intestinal epithelial cells (IECs) can sense these signals, integrate them, and interpret them for lamina propria lymphoid populations. One mechanism by which these signals are communicated is by the expression of non-classical major histocompatibility complex (MHC) class I molecules by IECs. Epithelial cells can express a surprising variety of non-classical MHC class I molecules. In some cases, IECs can act as non-professional antigen-presenting cells utilizing the expression of such non-classical MHC class I molecules to directly present bacterial antigens. In other cases, the expression of non-classical MHC class I molecules may act as a co-stimulatory molecule or adhesion molecule that can modify the mucosal immune response. Finally, the expression of these molecules on IECs can lead to a broad array of responses ranging from tolerance to inflammation. Overall, the IEC, via the expression of non-classical MHC class I molecules, is a central mediator of the constant crosstalk between the intestinal lumen and the mucosal immune system.


Spontaneous Expulsive Suprachoroidal Hemorrhage.
Spontaneous Expulsive Suprachoroidal Hemorrhage (SESCH) is an extremely rare phenomenon that leads to atraumatic spontaneous globe rupture. Only a handful of SESCH cases have been reported worldwide, primarily in developing countries. Risk factors associated with SESCH include uncontrolled hypertension, previous eye surgery, glaucoma, atherosclerosis, advanced age, and corneal damage. We report a case of expulsive suprachoroidal hemorrhage in a ninety-seven-year-old female. The patient presented to the emergency department with painful bleeding from her right eye. She denied any trauma or injury to the eye. She denied any use of anticoagulation. Physical examination showed a right ocular hematoma with mild active bleeding. She had exophthalmos and proptosis with extrusion of the ocular structures. A computed tomography scan of the orbits demonstrated right globe rupture with diffuse hemorrhage. Ophthalmology was consulted, and the patient subsequently had an evisceration of her right eye. WHY SHOULD AN EMERGENCY PHYSICIAN BE AWARE OF THIS?: Spontaneous expulsive choroidal hemorrhage is exceedingly rare. SESCH predominantly affects diseased eyes in the elderly. Early recognition of impending globe rupture in patients who present with suprachoroidal hemorrhage is necessary and requires prompt ophthalmology consultation. Emergency physicians must be aware that an atraumatic open globe can occur and requires immediate evaluation by an ophthalmologist and emergent repair in the operating suite.


Shortened time interval between colorectal cancer diagnosis and risk testing for hereditary colorectal cancer is not related to higher psychological distress.
Current diagnostic practices have shortened the interval between colorectal cancer (CRC) diagnosis and genetic analysis for Lynch syndrome by MSI-testing. We studied the relation of time between MSI-testing since CRC diagnosis (MSI-CRC interval) and psychological distress. We performed a cross-sectional study in 89 patients who had previously been treated for CRC. Data were collected during MSI-testing after genetic counseling. Psychological distress was measured with the IES, the SCL-90 and the POMS; social issues with the ISS, ISB and the ODHCF. The median time of MSI-CRC interval was 24 months (range 0-332), with 23% of the patients diagnosed less than 12 months and 42% more than 36 months prior to MSI-testing. In 34% of the patients cancer specific distress was high (IES scores >26). Mean psychopathology (SCL-90) scores were low, mean mood states (POMS) scores were moderate. Interval MSI-CRC was not related to psychological distress. High cancer specific distress was reported by 24% of patients diagnosed with CRC less than 12 months ago versus 39 and 35% by those diagnosed between 12 and 36 months and more than 36 months ago respectively. Distress was positively related to female gender (P = 0.04), religiousness (P = 0.01), low social support (P = 0.02) and difficulties with family communication (P < 0.001). Shortened time interval between CRC diagnosis and MSI-testing is not associated with higher psychological distress. Females, religious persons, those having low social support and those reporting difficulties communicating hereditary colorectal cancer with relatives are at higher risk for psychological distress.


[An experimental study on SRCA (subrenal capsule assay)--comparison of the test between normal and nude mice].
Six-day SRCA using normal mice developed by Bogden et al. is a promising in vivo chemosensitivity test. However, this method has a problem on the influence of the host reaction. We compared the tumor growth kinetics and host reaction between normal and nude mice. The tumor diameter increased until day 6 in normal and day 16 in nude mice, but the histological finding revealed many host reactive cells and few viable tumour cells on day 6 in normal mice, and well preserved tumour cells on day 16 in nude mice. These results were supported by flow cytometrical analysis. Autoradiogram using 3H-TdR showed a recovery of labeling index to the steady label by day 1. This index was similar between normal and nude mice. When antitumor activity of adriamycin, cisplatin or mitomycin C was compared with nude mice system, the order of effectiveness was the same as the system using nude mice implanted tumor cells subcutaneously and given the drugs intraperitoneally, but different in normal mice.


Protein kinase c-β-dependent activation of NF-κB in stromal cells is indispensable for the survival of chronic lymphocytic leukemia B cells in vivo.
Tumor cell survival critically depends on heterotypic communication with benign cells in the microenvironment. Here, we describe a survival signaling pathway activated in stromal cells by contact to B cells from patients with chronic lymphocytic leukemia (CLL). The expression of protein kinase C (PKC)-βII and the subsequent activation of NF-κB in bone marrow stromal cells are prerequisites to support the survival of malignant B cells. PKC-β knockout mice are insusceptible to CLL transplantations, underscoring the in vivo significance of the PKC-βII-NF-κB signaling pathway in the tumor microenvironment. Upregulated stromal PKC-βII in biopsies from patients with CLL, acute lymphoblastic leukemia, and mantle cell lymphoma suggests that this pathway may commonly be activated in a variety of hematological malignancies.


Pregnancy outcomes and menstrual function after fertility sparing surgery for pure ovarian dysgerminomas.
The medical records of 23 patients with pure ovarian dysgerminoma treated by conservative surgery at SSK Ankara Maternity Hospital between 1991 and 2001 were reviewed retrospectively. Pregnancy outcomes and menstrual function after conservative surgery were analyzed. Seven patients (30%) were incompletely staged and were assumed to be stage IA due to intraoperative findings. Fifteen patients (70%) were surgically staged. As a result 15 patients (65.2%) were diagnosed to be stage I while 2 patients (8.6%) were staged II and 6 patients (26%) were staged III. Wedge biopsy of contralateral ovary was performed in 17 patients (74%) and tumor was diagnosed in 2 (11.7%). Adjuvant chemotherapy was applied in 16 (70%) of the cases. BEP (bleomycin, etoposide, and cisplatin) was the most frequently used regimen (39%). Recurrences were seen in 3 (13%) patients. The sites of recurrences were contralateral ovary in all cases. Recurrences were treated with surgery and radiotherapy. None of the patients died of disease. Nine patients were sexually active. Five of these patients had prior live births. Three of the primiparous patients delivered after treatment. Five pregnancies occurred in 3 nulliparous patients at the time of primary surgery. Three of these reached term and healthy babies were delivered. One pregnancy was terminated at 18 weeks of gestation due to anencephaly and a dilatation and curettage was performed at 6th week in another patient. Menstrual problems occurred in 4 (17.3%) of the 23 patients. Fertility sparing treatments are safe in patients with pure ovarian dysgerminoma.


Amplified endogenous plasmin activity resolves acute thrombotic thrombocytopenic purpura in mice.
Essentials Plasmin is able to proteolyse von Willebrand factor. It was unclear if plasmin influences acute thrombotic thrombocytopenic purpura (TTP). Plasmin levels are increased during acute TTP though suppressed via plasmin(ogen) inhibitors. Allowing amplified endogenous plasmin activity in mice results in resolution of TTP signs. Background Thrombotic thrombocytopenic purpura (TTP) is an acute life-threatening pathology, caused by occlusive von Willebrand factor (VWF)-rich microthrombi that accumulate in the absence of ADAMTS-13. We previously demonstrated that plasmin can cleave VWF and that plasmin is generated in patients during acute TTP. However, the exact role of plasmin in TTP remains unclear. Objectives Investigate if endogenous plasmin-mediated proteolysis of VWF can influence acute TTP episodes. Results In mice with an acquired ADAMTS-13 deficiency, plasmin is generated during TTP as reflected by increased plasmin-α2-antiplasmin (PAP)-complex levels. However, mice still developed TTP, suggesting that this increase is not sufficient to control the pathology. As mice with TTP also had increased plasminogen activator inhibitor 1 (PAI-1) levels, we investigated whether blocking the plasmin(ogen) inhibitors would result in the generation of sufficient plasmin to influence TTP outcome in mice. Interestingly, when amplified plasmin activity was allowed (α2-antiplasmin-/- mice with inhibited PAI-1) in mice with an acquired ADAMTS-13 deficiency, a resolution of TTP signs was observed as a result of an increased proteolysis of VWF. In line with this, in patients with acute TTP, increased PAP-complex and PAI-1 levels were also observed. However, neither PAP-complex levels nor PAI-1 levels were related to TTP signs and outcome. Conclusions In conclusion, endogenous plasmin levels are increased during acute TTP, although limited via suppression through α2-antiplasmin and PAI-1. Only when amplified plasmin activity is allowed, plasmin can function as a back-up for ADAMTS-13 in mice and resolve TTP signs as a result of an increased proteolysis of VWF.


Freezing tolerance and avoidance in high tropical Andean plants: Is it equally represented in species with different plant height?
Freezing tolerance and avoidance were studied in several different sized species of the tropical high Andes (4200 m) to determine whether there was a relationship between plant height and cold resistance mechanisms. Freezing injury and supercooling capacity were determined in ground level plants (i.e. cushions, small rosettes and a perennial herb), intermediate height plants (shrubs and perennial herbs) and arborescent forms (i.e. giant rosettes and small trees). All ground-level plants showed tolerance as the main mechanism of resistance to cold temperatures. Arborescent forms showed avoidance mechanisms mainly through supercooling, while intermediate plants exhibited both. Insulation mechanisms to avoid low temperatures were present in the two extreme life-forms. We suggest that a combination of freezing tolerance and avoidance by insulation is least expensive and is a more secure mechanism for high tropical mountain plants than supercooling alone.


Transvascular endosonographic-guided needle biopsy of intrathoracic lesions.
Endoscopic ultrasound-guided biopsy techniques, including endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) and endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA), are currently the standard of care for the assessment of mediastinal lymphadenopathy. Traditionally, passing the needle through and through vascular structures has been avoided owing to the risk of bleeding. In this study, we evaluated the safety and diagnostic accuracy of transvascular endosonographic-guided biopsies of mediastinal, hilar and lung lesions. Our hypothesis is that the approach is safe and adds to the endoscopic armamentarium, avoiding the need for surgical biopsy in specifically selected cases. One hundred patients who underwent transvascular EBUS- or EUS-guided biopsy between 2012 and 2018 were identified from a prospective interventional endoscopy database. Biopsy was performed under EUS guidance in 65 patients and under EBUS guidance in 35 patients. The most frequent targets were the mediastinum (60 patients), lung (21 patients), and hilar lymph nodes (16 patients). The aorta was the vessel most commonly traversed (n = 57), followed by the pulmonary artery (n = 33). A median of 2 passes were performed per target (range, 1-5). The samples were adequate to make a diagnosis in 80 patients, and the endoscopic diagnosis was a malignancy in 62 patients. The overall sensitivity was 71.5%, and the accuracy was 74.5%. There were no observed intraoperative or immediate postoperative complications. A delayed complication, aortic pseudoaneurysm, was observed in 1 patient. Follow-up was completed in 84 patients, with a median duration of 12.3 ± 18 months. Transvascular endosonographic-guided biopsy is an important adjunct to conventional endoscopic techniques and allows the thoracic endoscopist to obtain biopsy specimens from intrathoracic lesions that are not accessible without vascular puncture.


Bright light therapy as a non-pharmacological treatment option for multiple sclerosis-related fatigue: A randomized sham-controlled trial.
Fatigue is a common symptom in people with multiple sclerosis (pwMS) that significantly impairs quality of life. Bright light therapy may be a cheap treatment option with little to no adverse events. To evaluate the effectiveness of bright light therapy as a treatment option for MS-related fatigue. This was randomized sham-controlled trial including 26 pwMS with a Fatigue Severity Scale (FSS) Score ≥36. Participants were assigned to receive either bright white light therapy (n = 13) or dim red light (sham-intervention; n = 13). Participants used the respective intervention for 30 min each morning for two weeks, followed by a two-week washout period. The primary endpoint was the difference in FSS scores following light treatment as calculated by analysis of covariance. There was no significant difference in FSS (F(1,23) = 2.39, p = .136, partial ⴄ2 = .094). However, FSS scores generally improved over the course of the study in a clinically relevant manner. Bright light therapy decreased FSS scores over the course of this study. However, this effect was not significant in comparison to a sham intervention.


Chromosomal aberrations in peripheral lymphocytes of cotton field workers exposed to pesticides.
Fifty-two pesticide sprayers in cotton fields were selected for the analysis of chromosomal aberrations in peripheral lymphocytes. Twenty-five samples were collected from healthy males who were not exposed to pesticides. Statistical analysis revealed that there was a significant increase in chromosomal aberrations in the exposed population compared to controls. Total chromosomal aberrations increased irrespective of duration of exposure.


Imaging Practice Patterns: Referral Network Analysis of a Single State of Origination.
The aim of this study was to examine the referral pattern of imaging studies requested in a single state compared with the potential location of interpretation. Analysis of Medicare patients in a DocGraph data set was performed to identify sequential different physician services claims for the same patient for which the second claim was for services provided by a radiologist. In the 2011 Medicare population, radiology referrals from physicians practicing in Georgia resulted in 76.5% of radiology interpretations by radiologists inside the state of Georgia. The states bordering Georgia accounted for 11.6% of interpretations in the Georgia market. The remaining interpretations were distributed throughout the remainder of the country. A significant proportion of routine imaging interpretation occurs outside the state in which an examination is performed. Additional studies are needed to identify complex drivers of imaging referral patterns, such as patient geographic location and demographics, radiologist workforce distribution, contractual obligations, and social relationships.


Macrophages disseminate pathogen associated molecular patterns through the direct extracellular release of the soluble content of their phagolysosomes.
Recognition of pathogen-or-damage-associated molecular patterns is critical to inflammation. However, most pathogen-or-damage-associated molecular patterns exist within intact microbes/cells and are typically part of non-diffusible, stable macromolecules that are not optimally immunostimulatory or available for immune detection. Partial digestion of microbes/cells following phagocytosis potentially generates new diffusible pathogen-or-damage-associated molecular patterns, however, our current understanding of phagosomal biology would have these molecules sequestered and destroyed within phagolysosomes. Here, we show the controlled release of partially-digested, soluble material from phagolysosomes of macrophages through transient, iterative fusion-fission events between mature phagolysosomes and the plasma membrane, a process we term eructophagy. Eructophagy is most active in proinflammatory macrophages and further induced by toll like receptor engagement. Eructophagy is mediated by genes encoding proteins required for autophagy and can activate vicinal cells by release of phagolysosomally-processed, partially-digested pathogen associated molecular patterns. We propose that eructophagy allows macrophages to amplify local inflammation through the processing and dissemination of pathogen-or-damage-associated molecular patterns.


Participant action research with bedside nurses to identify NANDA-International, Nursing Interventions Classification, and Nursing Outcomes Classification categories for hospitalized persons with diabetes.
Experienced bedside nurses identified 14 nursing diagnoses, 78 interventions, and 76 health outcomes for hospitalized persons with diabetes. Using these terms, the nursing department revised the standards of care and the electronic health record. Nurses' engagement in generating knowledge translated to increased interest in research. This methodology is recommended for other agencies.


A Solar cycle correlation of coronal element abundances in Sun-as-a-star observations.
The elemental composition in the coronae of low-activity solar-like stars appears to be related to fundamental stellar properties such as rotation, surface gravity, and spectral type. Here we use full-Sun observations from the Solar Dynamics Observatory, to show that when the Sun is observed as a star, the variation of coronal composition is highly correlated with a proxy for solar activity, the F10.7 cm radio flux, and therefore with the solar cycle phase. Similar cyclic variations should therefore be detectable spectroscopically in X-ray observations of solar analogs. The plasma composition in full-disk observations of the Sun is related to the evolution of coronal magnetic field activity. Our observations therefore introduce an uncertainty into the nature of any relationship between coronal composition and fixed stellar properties. The results highlight the importance of systematic full-cycle observations for understanding the elemental composition of solar-like stellar coronae.The Sun's elemental composition is a vital part of understanding the processes that transport energy from the interior to the outer atmosphere. Here, the authors show that if the Sun is observed as a star, then the variation of coronal composition is highly correlated with the F10.7cm radio flux.


Production of 2,3-dihydroxyisovalerate by Enterobacter cloacae.
2,3-Dihydroxyisovalerate is an intermediate of the valine synthesis pathway. However, neither natural microorganisms nor valine producing engineered strains have been reported yet to produce this chemical. Based on the 2,3-butanediol synthesis pathway, a biological route of 2,3-dihydroxyisovalerate production was developed using a budA and ilvD disrupted Klebsiella pneumoniae strain in our previous research. We hypothesised, that other 2,3-butanediol producing bacteria could be used for 2,3-dihydroxyisovalerate production. Here a budA disrupted Enterobacter cloacae was constructed, and this strain exhibited a high 2,3-dihydroxyisovalerate producing ability. Disruption of ilvD in E. cloacae ΔbudA further increased 2,3-dihydroxyisovalerate level. The disruption of budA, encoding an acetolactate decarboxylase, resulted in the acetolactate synthesized in the 2,3-butanediol synthesis pathway to flow into the valine synthesis pathway. The additional disruption of ilvD, encoding a dihydroxy acid dehydratase, prevented the 2,3-dihydroxyisovalerate to be further metabolized in the valine synthesis pathway. Thus, the disruption of both budA and ilvD in 2,3-butanediol producing strains might be an universal strategy for 2,3-dihydroxyisovalerate accumulation. After optimization of the medium components and culture parameters 31.2 g/L of 2,3-dihydroxyisovalerate was obtained with a productivity of 0.41 g/L h and a substrate conversion ratio of 0.56 mol/mol glucose in a fed-batch fermentation. This approach provides an economic way for 2,3-dihydroxyisovalerate production.


Sensitive detection of T4 polynucleotide kinase activity based on coupled exonuclease reaction and nicking enzyme-assisted fluorescence signal amplification.
As a prominent member of the 5'-kinase family, T4 polynucleotide kinase (PNK) plays an important role in gene function regulations, and the study of PNK activity and its potential inhibitors is significant for research related to the DNA phosphorylation process. Here, we proposed a novel strategy for the detection of PNK activity and its inhibition, which combines exonuclease enzyme reaction and nicking enzyme-assisted fluorescence signal amplification. Through recycling cleavage of DNA fluorescence probe for signal amplification, a highly sensitive PNK sensing platform is developed, and a very low detection limit of 0.05 mU/mL is achieved, which is better than or comparable to that of the previously reported PNK assays. The present approach adopts a simple separation-free procedure in which the enzyme assay is conducted in homogeneous solutions. Additionally, the inhibitory effects of several known kinase inhibitors on PNK have been successfully detected. Since the proposed assay exhibits the advantages of high sensitivity and simplicity, it holds great potential in providing a promising platform for convenient and highly sensitive detection of PNK activity and its inhibitors.


Final Overall Survival Analysis of the SOGUG Phase 2 MAJA Study: Maintenance Vinflunine Versus Best Supportive Care After First-Line Chemotherapy in Advanced Urothelial Carcinoma.
The MAJA study compared vinflunine (VFL) plus best supportive care (BSC) maintenance therapy versus BSC alone in advanced urothelial carcinoma responsive to first-line chemotherapy. The primary end point of progression-free survival was achieved. We present the final overall survival (OS) and long-term follow-up safety analyses. Patients were enrolled, and a subsequent post hoc analysis was performed on the basis of radiologic response or stabilization to first-line cisplatin/gemcitabine (CG) chemotherapy (4-6 cycles), according to Response Evaluation Criteria in Solid Tumors (RECIST). VFL + BSC versus BSC alone were randomly assigned until disease progression. At final analysis, 58 patients (66.7%) had died while 29 (33.3%) had survived; the BSC arm had higher mortality (VFL + BSC, n = 26, 59.1% vs. BSC, n = 32, 74.4%). Median follow-up of surviving patients was 38.8 months (interquartile range, 23.8-61.6). Median OS was 16.7 months (95% confidence interval, 0-34.5) in VFL and 13.2 months (95% confidence interval, 6-20.4) in the BSC groups (hazard ratio, 0.736; 95% confidence interval, 0.44-1.24, P = .182). Post hoc group division did not affect median OS in either study arm. Final analysis supported a benefit of VFL in maintenance therapy in patients with disease control after first-line treatment with CG, with no unexpected long-term adverse effects. The study was insufficiently powered to show a significant OS advantage.


Investigating genetic correlation and causality between nicotine dependence and ADHD in a broader psychiatric context.
People with attention-deficit/hyperactivity disorder (ADHD) or other psychiatric disorders show high rates of nicotine dependence (ND). This comorbidity might be (partly) explained by shared genetic factors. Genetic correlations between ND and ADHD (or other psychiatric disorders) have not yet been estimated. A significant genetic correlation might indicate genetic overlap, but could also reflect a causal relationship. In the present study we investigated the genetic correlation (with LD score regression analyses) between ND and ADHD, as well as between ND and other major psychiatric conditions (major depressive disorder, schizophrenia, anxiety, bipolar disorder, autism spectrum, anorexia nervosa, and antisocial behavior) based on the summary statistics of large Genome Wide Association studies. We explored the causal nature of the relationship between ND and ADHD using two-sample Mendelian randomization. We found a high genetic correlation between ND and ADHD (rg = .53, p = 1.85 × 10-13 ), and to a lesser extent also between ND-major depressive disorder (rg = .42, p = 3.6 × 10-11 ) and ND-schizophrenia (rg = .18, p = 1.1 × 10-3 ). We did not find evidence for a causal relationship from liability for ADHD to ND (which could be due to a lack of power). The strong genetic correlations might reflect different phenotypic manifestations of (partly) shared underlying genetic vulnerabilities. Combined with the lack of evidence for a causal relationship from liability for ADHD to ND, these findings stress the importance to further investigate the underlying genetic vulnerability explaining co-morbidity in psychiatric disorders.


Controlled release of bone morphogenetic protein (BMP)-2 from nanocomplex incorporated on hydroxyapatite-formed titanium surface.
Both osteoconductivity and osteoinductivity are equally very important aspects in a new bone formation and ultimately for bone regeneration. The purpose of this study was to create an environment, not only osteoconductive but also osteoinductive on titanium (Ti) surface. To do this bone morphogenetic protein-2 (BMP-2) nanocomplex (NC) was fabricated by using an ionic interaction between BMP-2 and chondroitin sulfate (CS). Meanwhile, Ti was chemically treated, then subjected to soaking in simulated body fluid (SBF), naming the sample Ti(C)-hydroxyapatite (HA). Once the BMP-2 NC was precipitated on the Ti(C)-HA surface, along with the addition of calcium/phosphate solution, the final product was formed as Ti(C)-HA-BMP-2. The size of NC was ranged from 150 to 250nm and the amount of CS was influential in determining both NC size and zeta potential. From the SEM observation, Ti surface was found nicely covered with the crystallized apatite layer that was identified using FTIR and NMR. The immobilized BMP-2 was released in a moderate rate for 4 weeks, without an initial burst of BMP-2. When mouse osteoblast cells were seeded on different Ti substrates, cell proliferation was faster in the Ti(C)-HA-BMP-2 group, as compared to other groups. The gene expression of bone-specific markers, osteocalcin and type I collagen, was significantly upregulated with the use of BMP-2 NC. The same result was witnessed in the measurement of alkaline phosphatase activity, in which the difference was statistically significant. This study demonstrated that the delivery system of BMP-2 NC was effective in holding BMP-2 on the apatite-coated Ti surface and that the Ti surface could be modified into the environment osteoinductive as well as osteoconductive.


Effect of aminophylline on high-energy phosphate metabolism and fatigue in the diaphragm.
Diaphragmatic fatigue causes respiratory failure, for which aminophylline has been used as therapy. Because the mechanism of action of aminophylline in reversing diaphragmatic fatigue is unclear, we used in vivo 31P magnetic resonance spectroscopy (MRS) to determine the relation between diaphragmatic activation, force output, and aerobic metabolism. Bilateral phrenic stimulation was used to pace the diaphragm in pentobarbital-anesthetized piglets (6-10 weeks old; n = 44). Esophageal and abdominal pressures were measured to calculate transdiaphragmatic pressure (Pdi) (Pdi = abdominal pressure-esophageal pressure) as an index of force output. Activation was determined by the amplitude of the compound action potential of the diaphragmatic electromyogram. Aerobic metabolism was assessed with a 31P MRS surface coil on the right hemidiaphragm with the animal in a 4.7-T magnet. The animals were divided into four groups based on aminophylline loading dose: saline, aminophylline 10 mg/kg (A10), aminophylline 20 mg/kg (A20), and aminophylline 40 mg/kg (A40). After aminophylline loading the diaphragm was paced for 25 min followed by a 10-min recovery. Aminophylline concentrations were 12.2 +/- 0.7, 21.9 +/- 2.4, and 44.9 +/- 3.6 mg/l in the A10, A20, and A40 groups, respectively. Compound action potential amplitude decreased in all groups by 30% after 25 min of pacing. Conversely, Pdi remained at 100 +/- 3% of the initial value after 5 min of pacing in the A40 group but decreased to 75 +/- 3% in the saline group. Pdi recovered completely (103 +/- 17%) in the A40 group but remained depressed (72 +/- 6%) in the saline group. Pdi values were intermediate in the A10 and A20 groups. MRS data revealed inadequate energy supply/demand ratio in the saline group such that the ratio of inorganic phosphate to phosphocreatine (Pi/PCr) increased to 1.01 +/- 0.09 after 5 min of pacing. Pi/PCr remained unchanged in the A40 group and was intermediate in the A10 and A20 groups. beta-Adenosine triphosphate and intracellular pH did not differ among groups or as a function of pacing. Diaphragmatic blood flow increased from a resting value of 35-60 to 300-410 ml.min-1 x 100 g-1 during pacing in all groups and was not affected by aminophylline dose. Aminophylline, in a dose-dependent fashion, delays the onset of fatigue and improves recovery from fatigue. Delayed fatigue is associated with improved aerobic metabolism as reflected in a low Pi/PCr ratio.


Directed Differentiation of Human Pluripotent Stem Cells into Inner Ear Organoids.
The sensory epithelia of the inner ear contain mechanosensitive hair cells that detect sound and head acceleration. This protocol details a 3D differentiation method to generate inner ear organoids containing sensory epithelia with hair cells. Human pluripotent stem cells are aggregated in low-binding 96-well plates and treated in chemically defined media with extracellular matrix to promote epithelialization. Small molecules and recombinant proteins are applied in a stepwise manner to recapitulate the morphogenic cues (BMP, TGF-β, FGF, and WNT) present during inner ear development in vivo. These treatments induce the sequential formation of nonneural ectoderm, otic-epibranchial progenitor domain, and otic placodes. The derived otic placodes then undergo self-guided morphogenesis to form otic vesicles, which eventually give rise to sensory epithelia containing hair cells and supporting cells, as well as neurons with synaptic formations to hair cells. This human stem cell-derived inner ear organoid system provides an ideal platform to study human inner ear development and disease in vitro.


Genetic architecture of body size in mammals.
Much of the heritability for human stature is caused by mutations of small-to-medium effect. This is because detrimental pleiotropy restricts large-effect mutations to very low frequencies.


The actions of in ovo cortisol on egg fertility, embryo development and the expression of growth-related genes in rainbow trout embryos, and the growth performance of juveniles.
Rainbow trout (Oncorhynchus mykiss) oocytes were incubated for 3 hr in ovarian fluid alone (CC), or cortisol-enriched ovarian fluid [100 or 1,000 ng ml(-1) (CL and CH, respectively)], after which they were fertilized; the growth and development of the embryos reared from these oocytes was monitored until first feed, and the juveniles were monitored for 9 months. The hatching rates of the CH group were significantly reduced, but the overall survival as measured at 40-week post-fertilization was similar in the three treatment groups. In addition, significant apparently biphasic changes relative to the CC group were found in the expression of some key growth-related genes in the CL and CH treatment groups, particularly IGF-1, IGF-2, GH1, GH2, GH receptors, and thyroid hormone receptors (TRα and TRβ). Moreover, the juveniles of the CL (but not the CH treatment group) exhibited enhanced growth; the enhanced growth could not be explained on the basis of increased feed conversion efficiency or changes in serum GH levels at the juvenile stage. Additionally, relative growth rates from the three treatment groups were similar, suggesting that the biphasic growth-enhancing effects of cortisol occurred very early in embryogenesis.


A microdialysis study of the in vivo release of 5-HT in the median raphe nucleus of the rat.
The present study has examined several characteristics of the release of 5-HT in the median raphe nucleus in terms of its dependence of nerve impulse, provenance of a vesicular storage fraction as well as the regulatory role played by 5-HT1A receptors. Tetrodotoxin (1 microM) and reserpine (5 mg kg(-1), i.p.) virtually suppressed the output of 5-HT. The administration of EEDQ (10 mg kg(-1), i.p.) did not alter the basal release of 5-HT but abolished the reduction of 5-HT release induced by 8-OH-DPAT (0.1 mg kg(-1), s.c.). The perfusion of 1-100 microM of 8-OH-DPAT or the novel 5-HT1A agonist BAY x 3702 decreased the efflux of 5-HT, whereas the perfusion of the 5-HT1A antagonist WAY-100635 failed to alter 5-HT release. The decrease in dialysate 5-HT induced by 100 microM 8-OH-DPAT was reversed by the concurrent perfusion of 100 microM WAY-100635. Also, the perfusion of 100 microM WAY-100635 for 2 h inhibited partly the reduction of 5-HT release evoked by the systemic administration of 8-OH-DPAT (0.1 mg kg(-1)). These results indicate that extracellular 5-HT in the median raphe nucleus is stored in vesicles and released in an impulse-dependent manner. Also, the basal release of 5-HT in the median raphe nucleus does not appear to be under the tonic control of somatodendritic 5-HT1A receptors by endogenous 5-HT. Instead, this feedback mechanism seems to be triggered when an excess of the transmitter or a 5-HT1A agonist is present in the extracellular space of the median raphe nucleus.


Spinal subarachnoid hemorrhage due to a filum terminale ependymoma.
We present a case of spinal subarachnoid hemorrhage due to an ependymoma of the filum terminale in a 23-year-old male. Clinical signs indicating a spinal origin of the subarachnoid hemorrhage are discussed. Subarachnoid hemorrhages are only rarely caused by an intraspinal tumor, most of which are located in the cauda equina. Our findings in this case proved the value of MRI examination in tumors of the cauda equina.


Partner smoking and maternal cotinine during pregnancy: implications for negative control methods.
Comparison of the associations of maternal and mother's partner smoking with offspring outcomes is, in theory, a useful method for assessing whether there may be an intrauterine effect of tobacco exposure on these outcomes. However, this approach assumes that the effects of passive smoking from exposure to partner smoking during pregnancy are minimal. We evaluated this assumption using a biochemical measure of tobacco exposure in pregnant women. Cotinine levels taken during the first trimester of pregnancy were measured in a sample of 3928 women from the Avon Longitudinal Study of Parents and Children. Median cotinine values were compared across categories of smoking heaviness (cigarettes per day) of the women during the first trimester and in non-smoking women by the smoking heaviness of their partner. Cotinine levels were substantially higher in women who smoked compared to non-smokers (range of medians across smoking heaviness categories: 900-5362 ng/ml versus 20 ng/ml, interquartile range (IQR) (0-63) for non-smokers). In contrast, cotinine levels in non-smoking women were only very weakly related to partner smoking status (range of medians in women with smoking partners: 34-69 ng/ml versus 12 ng/ml, IQR (0-48) in women with non-smoking partners). Levels of tobacco exposure from partner smoking, as assessed by cotinine, were low in non-smoking pregnant women. This suggests that using mother's partner's smoking as a negative control for investigating intrauterine effects is valid.


Investigating the optimal diagnostic value of histamine for diagnosing perioperative hypersensitivity: a prospective, observational study.
Although several guidelines recommend measuring blood tryptase and histamine concentrations to diagnose perioperative anaphylaxis (POA), tryptase measurement is more common. The appropriate timing of blood collection and the diagnostic threshold for histamine measurement are still controversial. To address these issues, histamine concentrations in patients with anaphylaxis and those with anaphylaxis-uncertain were compared in our previous study, the Japanese Epidemiologic Study for Perioperative Anaphylaxis (JESPA). However, because we could not rule out the possibility that the anaphylactic-uncertain group included anaphylactic patients, histamine concentrations were measured in patients who underwent general anesthesia with no complications as controls in the present study. Histamine levels were measured at anesthesia induction (baseline), 30 min (first point), and 2 h (second point) after the start of surgery in 30 control patients. Histamine concentrations in controls were lower than in patients with POA in JESPA at the first and second points. At the first point, a threshold of 1.5 ng/ml resulted in sensitivity of 77% and specificity of 100%. A threshold of 1.1 ng/ml at the second point resulted in sensitivity of 67% and specificity of 87%. Measurement of histamine concentrations within two hours after symptom onset might help diagnose POA.


CHAC1 as a novel biomarker for distinguishing alopecia from other dermatological diseases and determining its severity.
Alopecia Areata (AA) is characterised by an autoimmune response to hair follicles (HFs) and its exact pathobiology remains unclear. The current study aims to look into the molecular changes in the skin of AA patients as well as the potential underlying molecular mechanisms of AA in order to identify potential candidates for early detection and treatment of AA. We applied Weighted Gene Co-expression Network Analysis (WGCNA) to identify key modules, hub genes, and mRNA-miRNA regulatory networks associated with AA. Furthermore, Chi2 as a machine-learning algorithm was used to compute the gene importance in AA. Finally, drug-target construction revealed the potential of repositioning drugs for the treatment of AA. Our analysis using four AA data sets established a network strongly correlated to AA pathogenicity based on GZMA, OXCT2, HOXC13, KRT40, COMP, CHAC1, and KRT83 hub genes. Interestingly, machine learning introduced these genes as important in AA pathogenicity. Besides that, using another ten data sets, we showed that CHAC1 could clearly distinguish AA from similar clinical phenotypes, such as scarring alopecia due to psoriasis. Also, two FDA-approved drug candidates and 30 experimentally validated miRNAs were identified that affected the co-expression network. Using transcriptome analysis, suggested CHAC1 as a potential diagnostic predictor to diagnose AA.


Uncovering the influence of social skills and psychosociological factors on pain sensitivity using structural equation modeling.
Pain is a subjective emotional experience that is influenced by psychosociological factors such as social skills, which are defined as problem-solving abilities in social interactions. This study aimed to reveal the relationships among pain, social skills, and other psychosociological factors by using structural equation modeling. A total of 101 healthy volunteers (41 men and 60 women; mean age: 36.6±12.7 years) participated in this study. To evoke participants' sense of inner pain, we showed them images of painful scenes on a PC screen and asked them to evaluate the pain intensity by using the visual analog scale (VAS). We examined the correlation between social skills and VAS, constructed a hypothetical model based on results from previous studies and the current correlational analysis results, and verified the model's fit using structural equation modeling. We found significant positive correlations between VAS and total social skills values, as well as between VAS and the "start of relationships" subscales. Structural equation modeling revealed that the values for "start of relationships" had a direct effect on VAS values (path coefficient =0.32, p<0.01). In addition, the "start of relationships" had both a direct and an indirect effect on psychological factors via social support. The results indicated that extroverted people are more sensitive to inner pain and tend to get more social support and maintain a better psychological condition.


Blocking coprophagy increases the levels of inflammation and depression in healthy mice as well as mice receiving fecal microbiota transplantation from disease model mice donors.
Rodents have been extensively used as animal models in microbiome studies. However, all rodents have a habitual nature called coprophagy, a phenomenon that they self-reinoculate feces into their gastrointestinal tract. Recent studies have shown that blocking coprophagy can alter rodents' diversity of gut microbiota, metabolism, neurochemistry, and cognitive behavior. However, whether rodents' coprophagy behavior affects the levels of inflammation and depression is unclear. In order to address this problem, we first blocked coprophagy in healthy mice. It displayed an increase in the levels of depression, verified by depressive-like behaviors and mood-related indicators, and inflammation, verified by the increased levels of the pro-inflammatory cytokine, in coprophagy-blocked mice. Furthermore, we transplanted fecal microbiota from chronic restraint stress (CRS) depression model mice and lipopolysaccharide (LPS) inflammation model mice to healthy recipient mice, respectively. It showed that the disease-like phenotypes in the coprophagy-blocked group were worse than those in the coprophagy-unblocked group, including severer depressive symptoms and higher levels of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α and IFN-γ) in serum, prefrontal cortex (PFC), and hippocampus (HIP). These findings showed that blocking coprophagy in mice not only increased the levels of inflammation and depression in healthy mice but also aggravated inflammation and depression induced by fecal microbiota from disease donors. The discovery may provide a vital reference for future research involving FMT in rodents.


Characteristics and Adoption Success of Shelter Dogs Assessed as Resource Guarders.
Some domestic dogs aggressively guard resources. Canine resource guarding impacts public health through dog bites and affects dog welfare through adoption and euthanasia policies at animal shelters. However, little is known about the demographic characteristics and adoption success of dogs assessed as resource guarders during shelter behavioral evaluations. I reviewed nearly five years of records from a New York (NY) SPCA and categorized 1016 dogs by sex; age; size; reproductive status; and resource guarding. I then examined how these characteristics influenced the returns of dogs by adopters. The prevalence of resource guarding in this shelter dog population was 15%. Resource guarding was more common in adult and senior dogs than in juvenile dogs; and it was more common in small and large dogs than medium-sized dogs. Spayed females were more likely than intact females to guard food; neutered males and intact males did not differ in their likelihood of food guarding. Most dogs identified as resource guarders showed mild to moderate guarding. Severe guarders were more likely to be returned by adopters; although almost all were eventually re-adopted and not returned to the shelter. Data presented here provide the most comprehensive description of resource guarders in a shelter dog population and show the successful re-homing of most.


On the pathogenesis of chronic alcoholic pancreatitis from the viewpoint of experimental results in rats.
To elucidate the pathogenesis of chronic alcoholic pancreatitis, light-microscopic and electron microscopic observations of pancreases from Wistar strain rats given 20% ethanol ad lib, instead of water, were performed. Results were as follows: (1) In 3 (11%) rats, focal lobular unit acinar cell damage was observed under light-microscopy. In these cases, the intercalated ducts and centroacinar cells showed proliferation independently of protein plugs in ducts. (2) After ethanol intake, several degenerative changes in the acinar cells and ductular cells were observed under electron microscopy. The earliest and most consistent change was the irregular formation of microvilli and interdigitation between the parenchymal cells, accompanied by widening of the intercellular space, especially between the acinar and centroacinar cells. After three months of ethanol intake, an increase in the lysosomes and mitochondrial alteration occurred in acinar cells, and after six months of ethanol ingestion, there was tortuous ductular proliferation accompanied by the degeneration of the ductular cells. In addition to these parenchymal changes, the capillary endothelium and nervous tissue of the pancreas showed occasional degenerative changes after three to six months of ethanol intake. These results suggest that repeated degeneration and regeneration of the ductular cells results in deformation of ductular structures, with subsequent stenosis, and represents the primary event in the development of chronic alcoholic pancreatitis in rats.


Comparison between [18F]fluorination and [18F]fluoroethylation reactions for the synthesis of the PDE10A PET radiotracer [18F]MNI-659.
2-(2-(3-(4-(2-[18F]Fluoroethoxy)phenyl)-7-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)ethyl)-4-isopropoxyisoindoline-1,3-dione ([18F]MNI-659, [18F]1) is a useful PET radiotracer for imaging phosphodiesterase 10A (PDE10A) in human brain. [18F]1 has been previously prepared by direct [18F]fluorination of a tosylate precursor 2 with [18F]F-. The aim of this study was to determine the conditions for the [18F]fluorination reaction to obtain [18F]1 of high quality and with sufficient radioactivity for clinical use in our institute. Moreover, we synthesized [18F]1 by [18F]fluoroethylation of a phenol precursor 3 with [18F]fluoroethyl bromide ([18F]FEtBr), and the outcomes of [18F]fluorination and [18F]fluoroethylation were compared. We performed the automated synthesis of [18F]1 by [18F]fluorination and [18F]fluoroethylation using a multi-purpose synthesizer. We determined the amounts of tosylate precursor 2 and potassium carbonate as well as the reaction temperature for direct [18F]fluorination. The efficiency of the [18F]fluorination reaction was strongly affected by the amount of 2 and potassium carbonate. Under the determined reaction conditions, [18F]1 with 0.82±0.2GBq was obtained in 13.6%±3.3% radiochemical yield (n=8, decay-corrected to EOB and based on [18F]F-) at EOS, starting from 11.5±0.4GBq of cyclotron-produced [18F]F-. On the other hand, the [18F]fluoroethylation of 3 with [18F]FEtBr produced [18F]1 with 1.0±0.2GBq and in 22.5±2.5 % radiochemical yields (n=7, decay-corrected to EOB and based on [18F]F-) at EOS, starting from 7.4GBq of cyclotron-produced [18F]F-. Clearly, [18F]fluoroethylation resulted in a higher radiochemical yield of [18F]1 than [18F]fluorination. [18F]1 of high quality and with sufficient radioactivity was successfully radiosynthesized by two methods. [18F]1 synthesized by direct [18F]fluorination has been approved and will be provided for clinical use in our institute.


A Jeffrey Fluid Model for a Porous-walled Channel: Application to Flat Plate Dialyzer.
Creeping motion of a Jeffrey fluid in a small width porous-walled channel is presented with an application to flow in flat plate hemodialyzer. Darcy's law is used to characterize the fluid leakage through channel walls. Using suitable physical approximations, approximate analytical solution of equations of motion is obtained by employing perturbation method. Expressions for velocity field and the hydrostatic pressure are obtained. Effects of filtration coefficient, the inlet pressure and Jeffrey fluid parameters on the flow characteristics are discussed graphically. The derived results are used to study the flow of filtrate in a flat plat hemodialyzer. Using the derived solutions, theoretical values of the filtration rate and the mean pressure difference in the hemodialyzer are calculated. On comparing the computed results with the available experimental data, a reasonable agreement between the two is found. It is concluded that the presented model can be used to study the hydrodynamical aspects of the fluid flow in a flat plate hemodialyzer.


Bone-filling mesh container versus percutaneous kyphoplasty in treating Kümmell's disease.
Kümmell's disease (eponymous name for osteonecrosis and collapse of a vertebral body due to ischemia and non-union of anterior vertebral body wedge fractures after major trauma) cannot heal spontaneously. Bone-filling mesh container (BFMC) can significantly relieve pain, help the correction of kyphosis, and may prevent cement leakage. This pilot study may provide the basis for the design of future studies. To compare the effectiveness and safety of BFMC and percutaneous kyphoplasty (PKP) for treatment of Kümmell's disease. From August 2016 to May 2018, 40 patients with Kümmell's disease were admitted to Guizhou Provincial People's Hospital. Among them, 20 patients (20 vertebral bodies) received PKP (PKP group) and the other 20 received BFMC (BFMC group). Operation time, Visual Analogue Scale (VAS), Oswestry Disability Index (ODI), Cobb's angle changes, and related complications were recorded. All patients underwent operations successfully. VAS scores and ODI of both groups at each postoperative time point were lower than preoperatively, with statistically significant difference (p < 0.05). Postoperative Cobb's angle of both groups postoperatively was lower than preoperatively (p < 0.05). Cement leakage occurred in eight vertebrae (8/20) in the PKP group and in one vertebra (1/20) in the BFMC group. No complications such as pulmonary embolism, paraplegia, or perioperative death occurred during operation in both groups. Adjacent vertebral refractures occurred in five patients (5/20) in the PKP group and in four patients (4/20) in the BFMC group, with no significant difference in the incidence rate of refractures in both groups but the material is too small to verify statistically. Both PKP and BFMC technologies can significantly relieve pain and help the correction of kyphosis while treating Kümmell's disease. Moreover, the BMFC may prevent cement leakage.


Gender differences in prevalence and socioeconomic determinants of hypertension: findings from the WHO STEPs survey in a rural community of Vietnam.
In Vietnam, hypertension was estimated to cause a large number of deaths in hospitals. However, population-based knowledge about the magnitude of hypertension in Vietnam and its relationship with socioeconomic status, especially in the rural communities, still remains scarce. This paper, taking advantage of a study on noncommunicable disease (NCD) risk factors in Bavi district, Vietnam, using the WHO STEPs approach, estimates the prevalence of hypertension in the setting and examines its association with some socioeconomic factors. A representative sample comprising 2000 adults aged 25-64 years were selected randomly and surveyed in 2002. The JNC VII criteria for hypertension were used. Socioeconomic status of the study subjects was estimated by assessing their education, occupation and economic conditions. Descriptive techniques and multivariate logistic regression were used. The prevalence of hypertension was 14.1%. Of hypertensives, only 17.4% were aware of their hypertensive status. Men were hypertensive more often than women and age was positively associated with hypertension. The association between hypertension and socioeconomic status was complex and differed between men and women. Among men, those with lower educational and occupational status but who were richer were more likely to be hypertensive. More women with lower occupational and economic status were hypertensive.


"[It] is now my responsibility to fulfill that wish:" Clinical and rapid autopsy staff members' experiences and perceptions of HIV reservoir research at the end of life.
Little is known about the effects of HIV reservoir research at the end of life on staff members involved. Staff members' perceptions and experiences were assessed related to their involvement in the Last Gift, a rapid autopsy study at the University of California San Diego enrolling people living with HIV who are terminally ill and have a desire to contribute to HIV cure-related research. Two focus group discussions consisting of clinical (n = 7) and rapid research autopsy (n = 8) staff members were conducted to understand the perspectives of staff members and the impact the Last Gift rapid autopsy study had on them. The total sample consisted of 66.7% females and 33.3% males and was ethnically diverse (66.7% Caucasian, 6.7% African American, 20.0% Asian descent, 6.7% Hispanic descent and American Indian) with a range of experience in the HIV field from 1 year to 30 years. Qualitative focus group data revealed five major themes underlying study staff members' multilayered mental and practical involvement: 1) positive perceptions of the Last Gift study, with sub-themes including Last Gift study participants' altruism, fulfillment, and control at the end of life, 2) perceptions of staff members' close involvement in the Last Gift study, with sub-themes related to staff members' cognitive processing, self-actualization and fulfillment, stress management and resilience, coping mechanisms, and gratitude toward Last Gift participants and toward the study itself, 3) considerations for successful and sustainable study implementation, such as ethical awareness and sustained community and patient engagement, 4) collaborative learning and organizational processes and the value of interdependence between staff members, and 5) considerations for potential study scale-up at other clinical research sites. Understanding staff members' nuanced emotional and procedural experiences is crucial to the Last Gift study's sustainability and will inform similar cure research studies conducted with people living with HIV at the end of life. The study's potential reproducibility depends on a robust research infrastructure with established, interdependent clinical and rapid autopsy teams, continuous community engagement, and an ethical and well-informed engagement process with people living with HIV.


Spatiotemporally mapping temperature dynamics of lysosomes and mitochondria using cascade organelle-targeting upconversion nanoparticles.
The intracellular metabolism of organelles, like lysosomes and mitochondria, is highly coordinated spatiotemporally and functionally. The activities of lysosomal enzymes significantly rely on the cytoplasmic temperature, and heat is constantly released by mitochondria as the byproduct of adenosine triphosphate (ATP) generation during active metabolism. Here, we developed temperature-sensitive LysoDots and MitoDots to monitor the in situ thermal dynamics of lysosomes and mitochondria. The design is based on upconversion nanoparticles (UCNPs) with high-density surface modifications to achieve the exceptionally high sensitivity of 2.7% K-1 and low uncertainty of 0.8 K for nanothermometry to be used in living cells. We show the measurement is independent of the ion concentrations and pH values. With Ca2+ ion shock, the temperatures of both lysosomes and mitochondria increased by ∼2 to 4 °C. Intriguingly, with chloroquine (CQ) treatment, the lysosomal temperature was observed to decrease by up to ∼3 °C, while mitochondria remained relatively stable. Lastly, with oxidative phosphorylation inhibitor treatment, we observed an ∼3 to 7 °C temperature increase and a thermal transition from mitochondria to lysosomes. These observations indicate different metabolic pathways and thermal transitions between lysosomes and mitochondria inside HeLa cells. The nanothermometry probes provide a powerful tool for multimodality functional imaging of subcellular organelles and interactions with high spatial, temporal, and thermal dynamics resolutions.


Markerless Analysis of Articulatory Movements in Patients With Parkinson's Disease.
A large percentage of patients with Parkinson's disease have hypokinetic dysarthria, exhibiting reduced peak velocities of jaw and lips during speech. This limitation implies a reduction of speech intelligibility for such patients. This work aims at testing a cost-effective markerless approach for assessing kinematic parameters of hypokinetic dysarthria. Kinematic parameters of the lips are calculated during a syllable repetition task from 14 Parkinsonian patients and 14 age-matched control subjects. Combining color and depth frames provided by a depth sensor (Microsoft Kinect), we computed the three-dimensional coordinates of main facial points. The peak velocities and accelerations of the lower lip during a syllable repetition task are considered to compare the two groups. Results show that Parkinsonian patients exhibit reduced peak velocities of the lower lip, both during the opening and the closing phase of the mouth. In addition, peak values of acceleration are reduced in Parkinsonian patients, although with significant differences only in the opening phase with respect to healthy control subjects. The novel contribution of this work is the implementation of an entirely markerless technique capable to detect signs of hypokinetic dysarthria for the analysis of articulatory movements during speech. Although a large number of Parkinsonian patients have hypokinetic dysarthria, only a small percentage of them undergoes speech therapy to increase their articulatory movements. The system proposed here could be easily implemented in a home environment, thus, increasing the percentage of patients who can perform speech rehabilitation at home.


Energy-related parameters and their association with age, gender, and morphometric measurements in healthy donkeys.
Donkeys are commonly afflicted by endocrine and metabolic disturbances but few studies have investigated endocrine variables involved in energy regulation and their association with morphometric indices, age or gender in this species. Hemostatic and clinical differences have been demonstrated between horses and donkeys, so to consider both species as metabolically and endocrinologically similar could lead to misdiagnosis. In this study, plasma concentrations of glucose, triglycerides and endocrine factors involved in energy homeostasis (insulin, glucagon, leptin, adiponectin, ghrelin and insulin-like growth factor [IGF]-1) were measured and their association with morphometric variables (body condition score, neck scoring and body mass index), gender and age was determined in 62 healthy donkeys. In addition, a neck scoring system specific for donkeys was developed. Insulin, glucagon, leptin and IGF-1 concentrations were found to be similar between donkeys and other species, but adiponectin and active ghrelin were lower in donkeys than horses. Donkeys with larger neck scores and body mass indices had higher triglyceride, leptin and IGF-1 concentrations. A sexual dimorphism was observed on all morphometric measurements and plasma glucose concentrations independent of adiposity. Younger animals had lower morphometric measurements and triglyceride and leptin concentrations.


Hydroxyethyl starch-induced pruritus relieved by a combination of menthol and camphor.
Hydroxyethyl starch is a key component of many colloid volume expanders used in hypovolemic shock and otologic disease. Pruritus is a common side effect. Histopathology reveals multiple cytoplasmic vacuoles in dermal macrophages, endothelial cells, and perineural cells with electron-dense foreign material within the said vacuoles. Although classically refractory to treatment with corticosteroids and antihistamines, some benefit has been achieved with capsaicin, ultraviolet light therapy, and oral naltrexone. We present a case responsive to menthol and camphor and discuss the possible therapeutic mechanism.


An ideal weed: plasticity and invasiveness in Polygonum cespitosum.
The introduced Asian plant Polygonum cespitosum has only recently become invasive in northeastern North America, spreading into sunny as well as shaded habitats. We present findings from a multiyear case study of this ongoing species invasion, drawing on field environmental measurements, glasshouse plasticity and resurrection experiments, and molecular genetic (microsatellite) data. We focus in particular on patterns of individual phenotypic plasticity (norms of reaction), their diversity within and among populations in the species' introduced range, and their contribution to its potential to evolve even greater invasiveness. Genotypes from introduced-range P. cespitosum populations have recently evolved to express greater adaptive plasticity to full sun and/or dry conditions without any loss of fitness in shade. Evidently, this species may evolve the sort of "general-purpose genotypes" hypothesized by Herbert Baker to characterize an "ideal weed." Indeed, we identified certain genotypes capable of extremely high reproductive output across contrasting conditions, including sunny, shaded, moist, and dry. Populations containing these high-performance genotypes had consistently higher fitness in all glasshouse habitats; there was no evidence for local adaptive differentiation among populations from sunny, shaded, moist, or dry sites. Norm of reaction data may provide valuable insights to invasion biology: the presence of broadly adaptive, high-performance genotypes can promote a species' ecological spread while providing the fuel for increased invasiveness to evolve.


Assessment of antimalarial effect of ICL670A on in vitro cultures of Plasmodium falciparum.
We tested in vitro the antimalarial properties of ICL670A, a newly developed iron chelator for the long-term oral treatment of iron overload. Ring-stage synchronized cultures of Plasmodium falciparum cultured in human erythrocytes were exposed to different concentrations of ICL670A and the conventional iron chelator, desferrioxamine B (DFO), for 48 h. Malarial growth was measured by incorporation of [3H]-hypoxanthine. ICL670A at 30 micromol/l had marked antimalarial activity that was observable by 6 h after beginning the exposure of ring-stage parasites to the agent. Over 48 h of culture, malarial growth was significantly lower with ICL670A than with DFO at concentrations of both 30 micromol/l (P = 0.008) and 60 micromol/l (P = 0.001). At 48 h, growth relative to control was 53% with ICL670A and 83% with DFO at concentrations of 30 micromol/l, and 20% with ICL670A and 26% with DFO at concentrations of 60 micromol/l. Standard 50% inhibitory concentrations (IC50s) were similar for ICL670A and DFO. Precomplexation with iron completely abolished the inhibitory effect of ICL670A, indicating that this new agent, like DFO, probably inhibits parasite growth via deprivation of iron from critical targets within the parasite. Further studies to address the question of the antimalarial potential of ICL670A in combination with classic antimalarials would be of interest.


Synthesis and functionalization of silica-based nanoparticles with fluorescent biocompounds extracted from Eysenhardtia polystachya for biological applications.
Several types of dyes or fluorophores are used for the detection of interactions between drug carriers and cells, within biomedicine field. However, many of them have a certain level of toxicity and instability affecting their biological properties. Different studies have demonstrated that nanoparticles (NPs) have interesting properties that could be used to stabilize diverse biomolecules, including dyes. Here, we report the synthesis of a novel nanosystem by the functionalization of silica NPs using biocompounds extracted from Mexican tree "Palo azul" (Eysenhardtia polystachya) and APTES as a coupling agent. Particle size, electrical properties, and morphology of the novel nanosystem were analyzed. The extracted biocompounds presented fluorescence which prevails over time, even after nanosystem formation and apparent cellular internalization. These were detected using MCF-7 cells visualized by confocal laser-scanning microscopy (CLSM), finding that the nanosystem was able to internalize into cells and act as a fluorescent biomarker. By this method, our novel nanosystem opens the possibilities to obtain sensitive data in a noninvasive manner for biological applications, such as early-stage cancer diagnosis, drug delivery, and pathogen detection.


Photo Rounds: Painful facial blisters, fever, and conjunctivitis.
Following Tx for facial blisters, our patient returned with what appeared to be viral conjunctivitis. Further evaluation revealed a missed tip-off to the proper Dx.


Carbonic anhydrase 4 disruption decreases synaptic and behavioral adaptations induced by cocaine withdrawal.
Cocaine use followed by withdrawal induces synaptic changes in nucleus accumbens (NAc), which are thought to underlie subsequent drug-seeking behaviors and relapse. Previous studies suggest that cocaine-induced synaptic changes depend on acid-sensing ion channels (ASICs). Here, we investigated potential involvement of carbonic anhydrase 4 (CA4), an extracellular pH-buffering enzyme. We examined effects of CA4 in mice on ASIC-mediated synaptic transmission in medium spiny neurons (MSNs) in NAc, as well as on cocaine-induced synaptic changes and behavior. We found that CA4 is expressed in the NAc and present in synaptosomes. Disrupting CA4 either globally, or locally, increased ASIC-mediated synaptic currents in NAc MSNs and protected against cocaine withdrawal-induced changes in synapses and cocaine-seeking behavior. These findings raise the possibility that CA4 might be a previously unidentified therapeutic target for addiction and relapse.


Dual-energy CT evaluation of gastrointestinal bleeding.
Gastrointestinal bleeding is a common cause for hospital admissions and is an important cause of morbidity and mortality. Although endoscopy is accepted as the standard initial diagnostic modality for the evaluation of gastrointestinal bleeding, multiphasic computed tomography (CT) imaging has become an alternative diagnostic tool. Dual-energy CT with post-processing techniques may have additional advantages over single-energy computed tomography in evaluation of gastrointestinal bleeding. In this article, we discuss the role of dual-energy CT in the evaluation of gastrointestinal bleeding with potential advantages over conventional CT and limitations.


Natural and synthetic drugs and formulations for intravaginal HPV clearance.
Except for a few preventative Human Papillomavirus (HPV) vaccines, there is currently no cure for HPV infection. There are a number of cutting-edge strategies and potent medications or herbal formulations that can be applied topically for early clearance of HPV infection before HPV DNA gets integrated into host cell genome. This is facilitated due to cervical cancer having distinct and well-recognized long precancerous stages. This review aims to outline every possible medication and formulation, both natural and synthetic, that can be applied topically as intravaginal application to help remove HPV infection at an early precancerous stage. Several anti-HPV/HPV clearance compounds and formulations for high-grade lesions are undergoing clinical trials. However, the majority of compounds are still in the early stages of development and require additional research to become viable HPV clearance candidates. Synthetic drugs may be more promising because they may have a more targeted effect; however, they may also have significant adverse effects. On the other hand, natural medications are safer to use. They are less specific, but have minimal to no adverse effects. This article may serve as a valuable resource of information for managing and preventing precancerous carcinogenic HPV infections. Research could be directed toward developing candidate drugs to make evidence-based decisions about advancing them to clinical trials and, eventually, to the market for potential use in the prevention and control of cervical cancer, which is almost always preventable or even curable if detected early.


Strategies facilitating practice change in pediatric cancer: a systematic review.
By conducting a systematic review, we describe strategies to actively disseminate knowledge or facilitate practice change among healthcare providers caring for children with cancer and we evaluate the effectiveness of these strategies. We searched Ovid Medline, EMBASE and PsychINFO. Fully published primary studies were included if they evaluated one or more professional intervention strategies to actively disseminate knowledge or facilitate practice change in pediatric cancer or hematopoietic stem cell transplantation. Data extracted included study characteristics and strategies evaluated. In studies with a quantitative analysis of patient outcomes, the relationship between study-level characteristics and statistically significant primary analyses was evaluated. Of 20 644 titles and abstracts screened, 146 studies were retrieved in full and 60 were included. In 20 studies, quantitative evaluation of patient outcomes was examined and a primary outcome was stated. Eighteen studies were 'before and after' design; there were no randomized studies. All studies were at risk for bias. Interrupted time series was never the primary analytic approach. No specific strategy type was successful at improving patient outcomes. Literature describing strategies to facilitate practice change in pediatric cancer is emerging. However, major methodological limitations exist. Studies with robust designs are required to identify effective strategies to effect practice change.


POTE protein, a cancer-testis antigen, is highly expressed in spermatids in human testis and is associated with apoptotic cells.
The POTE gene family encodes very closely related proteins that are highly expressed in testis and in many cancers. Recent studies indicate that the POTE proteins have a pro-apoptotic function. To examine if POTE is associated with cells that are undergoing apoptosis in testis, we determined the cellular location of POTE and of Cleaved Caspase-3 in testicular tissues from 26 azoospermic men. We found intense expression of POTE in round spermatids that are undergoing apoptosis, which are positive for Cleaved Caspase-3. This study suggests POTE may have a role in apoptosis in the human testis.


Isolation and identification of Acanthamoeba species from thermal spring environments in southern Taiwan.
Acanthamoeba species are free-living amoebae found in a range of environments. Within this genus, a number of species are recognized as human pathogens, potentially causing Acanthamoeba keratitis, granulomatous amoebic encephalitis, and chronic granulomatous lesions. In this study, 60 water samples were taken from four thermal spring recreation areas in southern Taiwan. We detected living Acanthamoeba spp. based on culture-confirmed detection combined with the molecular taxonomic identification method. Living Acanthamoeba spp. were detected in nine (15%) samples. The presence or absence of Acanthamoeba spp. in the water samples depended significantly on the pH value. The most frequently identified living Acanthamoeba genotype was T15 followed by T4, Acanthamoeba spp., and T2. Genotypes T2, T4, and T15 of Acanthamoeba, are responsible for Acanthamoeba keratitis as well as granulomatous amoebic encephalitis, and should therefore be considered a potential health risk associated with human activities in thermal spring environments.


Utility of Cell-Free DNA Detection in Transplant Oncology.
Transplant oncology is an emerging field in cancer treatment that applies transplant medicine, surgery, and oncology to improve cancer patient survival and quality of life. A critical concept that must be addressed to ensure the successful application of transplant oncology to patient care is efficient monitoring of tumor burden pre-and post-transplant and transplant rejection. Cell-free DNA (cfDNA) detection has emerged as a vital tool in revolutionizing the management of cancer patients who undergo organ transplantation. The advances in cfDNA technology have provided options to perform a pre-transplant evaluation of minimal residual disease (MRD) and post-transplant evaluation of cancer recurrence and transplant rejection. This review aims to provide a comprehensive overview of the history and emergence of cfDNA technology, its applications to specifically monitor tumor burden at pre-and post-transplant stages, and evaluate transplant rejection.


Adaptive conjoint analysis to determine perceived risk factors of farmers, veterinarians and AI technicians for introduction of BHV1 to dairy farms.
A study was carried out to determine the possibility of a more-closed farming system for (Dutch) dairy farms. The objective of the study was to provide effective and economically profitable management advice for improving the animal-health status of farms. Management measures will only be successfully applied if supported by farmers and their advisors (such as veterinarians). Therefore, the perception of farmers and advisors of the importance of various risk factors for the introduction of diseases to a farm was determined by using bovine herpes virus type 1 (BHV1) as an example. As part of the study, an evening-long workshop was organized and run thrice. In total, 49 farmers, veterinarians and AI technicians participated in these workshops. The computerized questionnaire technique was based on adaptive conjoint analysis (ACA). ACA has the advantage that participants can work with a large number of risk factors in a relatively short period of time. Another advantage of ACA (compared with standard questionnaires) is that the answers from each participant can be checked with regard to consistency with respect to the importance assigned to them. Data from participants with inconsistent responses can be excluded from further analyses. The results of the ACA interview were compared with the risk factors reported in the literature as being associated with BHV1 status (e.g. purchase of cattle, participation in cattle shows) and with farmers' actual management to prevent the introduction of diseases. The workshop participants were all operating in the dairy sector and they seemed well aware of the risk of direct animal contacts for the introduction of BHV1. Farmers thought visitors to be more risky than did AI technicians and (especially) veterinarians. Farmers who purchased cattle or participated in cattle shows were of the opinion that the risks of direct animal contacts were more important than did farmers who were not involved in those practices. Farmers whose farms were BHV1-positive (and participated in cattle shows more often) thought the risk of participation smaller than did farmers with BHV1-negative farms.


Toxicity of melanin-free ink of Sepia officinalis to transformed cell lines: identification of the active factor as tyrosinase.
The melanin-free ink of the cephalopod Sepia officinalis is shown to contain a heat labile proteinaceous component toxic to a variety of cell lines, including PC12 cells. Gel filtration chromatography indicated that the toxic component was concentrated in those fractions eluted at a molecular weight higher than 100 kDa and exhibiting the highest tyrosinase activity. SDS-PAGE analysis of the active fractions displayed a single major band migrating at an approximate molecular weight of 100 kDa, identical with that of the single tyrosinase band in the melanin-free ink. These data unambiguously demonstrated the identity of the toxic component with tyrosinase. Treatment of purified Sepia as well as of mushroom tyrosinase with an immobilized version of proteinase K resulted in a parallel loss of tyrosinase activity and cytotoxicity. Sepia apotyrosinase was ineffective in inducing cytotoxicity in PC12 cells. Purified Sepia tyrosinase was found to induce a significant increase in caspase 3 activity in PC12 cells, leading eventually to an irreversible apoptotic process. Overall, these results disclose a hitherto unrecognized property of tyrosinase that may lead to a reappraisal of its biological significance beyond that of a mere pigment producing enzyme.


The effect of substrate on the uptake of oxygen and sulphate ions by heat-damaged skin.
Respiration and sulphate uptake have been measured in guinea-pig skin subjected to heat damage and maintained on a medium containing sodium succinate. Succinate was found to have a protective effect against loss of oxygen uptake, a higher temperature being required for 50% effect than that reported with glucose as substrate. The protective effect did not extend to sulphate incorporation, suggesting that this process is itself heat-labile and that its failure with heat damage is not directly caused by diminution of the energy supply.


Distinctive clinicopathological features and KRAS and IDH1/2 mutation status of cholangiolocellular carcinoma.
Cholangiolocellular carcinoma (CLC) is classified as a subtype of combined hepatocellular cholangiocarcinoma with stem-cell features (CHC-SC) in the latest World Health Organization classification. This subclassification of CHC-SCs is controversial and the relevance of such classification is unclear. We analyzed a series of CHC-SCs and intrahepatic cholangiocarcinoma (iCCA) to clarify the clinicopathological features and mutational status of each tumor. Background liver disease, fibrosis stage, microvascular invasion, nodal metastasis, and IDH1/2 mutation status were associated with their histology. Compared with the intermediate cell subtype of CHC-SC (CHCs-SC-int), CLCs were less frequently associated with chronic viral hepatitis, and showed lower levels of serum alpha-fetoprotein. Compared with iCCAs, CLCs showed lower levels of serum carbohydrate antigen 19-9 (CA19-9) and a lower frequency of expression of S100P. Patients with iCCA showed worse overall survival than those with CLC or CHC-SC-int. In patients with iCCA, CLC, or CHC-SC-int, a histology of iCCA, microvascular invasion, and serum CA19-9 value of >100 U/mL were significant poor prognostic factors for overall survival in univariate analysis. Multivariate analysis showed that a high serum CA19-9 value was an independent poor prognostic factor for overall survival. Patients with CLC are likely to have a different etiology and mutational background from those with CHC-SC-int. Their clinicopathological manifestations are also different from those with classic iCCA. Our results suggest that CLC might be a distinct entity among primary liver carcinomas.


The Effect of GS-548351 on the Pharmacokinetics of Midazolam Following Multiple Doses of ANS-6637 in Healthy Adults.
ANS-6637, a pro-drug of GS-548351, is a selective, reversible inhibitor of aldehyde dehydrogenase isoform 2 under development as an anticraving agent for the treatment of substance use disorders. In vitro testing indicates that GS-548351 is an inhibitor and inducer of cytochrome P450 family 3, subfamily A (CYP3A). In this phase 1 single-center, open-label, fixed-sequence drug-drug interaction study we assessed the impact of steady-state GS-548351 on single-dose pharmacokinetics of midazolam, an index substrate for CYP3A. Twelve healthy volunteers received 600 mg of ANS-6637 by mouth daily from study days 3 to 8 and a single 5-mg oral dose of midazolam on days 1 and 8. Pharmacokinetic samples were collected over 24 hours on days 1 and 8, then analyzed using liquid chromatography-tandem mass spectrometry. The prespecified no-effect range for the 90% confidence interval (CI) of the geometric mean ratio (GMR) of midazolam coadministered with ANS-6637 (day 8) compared with midazolam alone (day 1) was 0.7-1.43. There was an increase in midazolam AUC0-∞ (GMR [90%CI]) that was within the no-effect range (1.26 [1.12-1.425]) and an increase in midazolam Cmax that was outside the range (1.22 [1.03-1.45]). The AUC0-∞ (1.08 [0.91-1.27]) and Cmax (0.95 [0.75-1.2]) of 1-hydroxymidazolam, the primary metabolite of midazolam, were also within the no-effect range. A single grade 3 adverse event (alanine aminotransferase elevation) was identified and resolved following discontinuation of the study drug. Overall, multidose ANS-6637 was well tolerated and did not alter the PK of midazolam beyond a small increase in AUC0-∞ that is unlikely to be clinically significant.


Constructing bi-plots for random forest: Tutorial.
Current technological developments have allowed for a significant increase and availability of data. Consequently, this has opened enormous opportunities for the machine learning and data science field, translating into the development of new algorithms in a wide range of applications in medical, biomedical, daily-life, and national security areas. Ensemble techniques are among the pillars of the machine learning field, and they can be defined as approaches in which multiple, complex, independent/uncorrelated, predictive models are subsequently combined by either averaging or voting to yield a higher model performance. Random forest (RF), a popular ensemble method, has been successfully applied in various domains due to its ability to build predictive models with high certainty and little necessity of model optimization. RF provides both a predictive model and an estimation of the variable importance. However, the estimation of the variable importance is based on thousands of trees, and therefore, it does not specify which variable is important for which sample group. The present study demonstrates an approach based on the pseudo-sample principle that allows for construction of bi-plots (i.e. spin plots) associated with RF models. The pseudo-sample principle for RF. is explained and demonstrated by using two simulated datasets, and three different types of real data, which include political sciences, food chemistry and the human microbiome data. The pseudo-sample bi-plots, associated with RF and its unsupervised version, allow for a versatile visualization of multivariate models, and the variable importance and the relation among them.


Loneliness and psychological distress before and during the COVID-19 pandemic: Relationships with social media identity bubbles.
Impacts of the COVID-19 pandemic on well-being and mental health are a concern worldwide. This article is based on two longitudinal studies that investigated the role of social media use in loneliness and psychological distress before and during the COVID-19 pandemic. Study 1 utilized nationally representative 3-point longitudinal data (n = 735) collected in 2017-2020 on the Finnish population. Study 2 utilized 5-point longitudinal data (n = 840) collected in 2019-2021 representing the Finnish working population. We analyzed the data using multilevel mixed-effects regression analysis. A longitudinal analysis of Study 1 showed that perceived loneliness did not increase among the Finnish population during the COVID-19 pandemic. Stronger involvement in social media identity bubbles predicted lower loneliness during the pandemic. Study 2 results showed that since the outbreak of the pandemic, psychological distress has increased among lonely individuals but not among the general working population. Involvement in social media identity bubbles predicted generally lower psychological distress during the COVID-19 pandemic, but it did not buffer against higher psychological distress among lonely individuals. The findings suggest that perceived loneliness is a risk factor for prolonged negative mental health effects of the pandemic. Social media identity bubbles can offer meaningful social resources during times of social distancing but cannot protect against higher psychological distress among those who perceive themselves as often lonely.


Radiosensitizer Effect of β-Apopicropodophyllin against Colorectal Cancer via Induction of Reactive Oxygen Species and Apoptosis.
β-apopicropodophyllin (APP), a derivative of podophyllotoxin (PPT), has been identified as a potential anti-cancer drug. This study tested whether APP acts as an anti-cancer drug and can sensitize colorectal cancer (CRC) cells to radiation treatment. APP exerted an anti-cancer effect against the CRC cell lines HCT116, DLD-1, SW480, and COLO320DM, with IC50 values of 7.88 nM, 8.22 nM, 9.84 nM, and 7.757 nM, respectively, for the induction of DNA damage. Clonogenic and cell counting assays indicated that the combined treatment of APP and γ-ionizing radiation (IR) showed greater retardation of cell growth than either treatment alone, suggesting that APP sensitized CRC cells to IR. Annexin V-propidium iodide (PI) assays and immunoblot analysis showed that the combined treatment of APP and IR increased apoptosis in CRC cells compared with either APP or IR alone. Results obtained from the xenograft experiments also indicated that the combination of APP and IR enhanced apoptosis in the in vivo animal model. Apoptosis induction by the combined treatment of APP and IR resulted from reactive oxygen species (ROS). Inhibition of ROS by N-acetylcysteine (NAC) restored cell viability and decreased the induction of apoptosis by APP and IR in CRC cells. Taken together, these results indicate that a combined treatment of APP and IR might promote apoptosis by inducing ROS in CRC cells.


The transfer of ketorolac tromethamine from maternal to foetal blood.
Thirty two women who were participating in an efficacy study comparing 10 mg ketorolac with 50 mg or 100 mg of pethidine in the relief of labour pain, underwent sampling of vein blood, for determination of plasma ketorolac concentrations. The sample was withdrawn at delivery and a sample of umbilical cord blood was withdrawn at the same time. The ratio of ketorolac concentrations in the cord blood sample: the maternal venous sample were calculated and plotted against the time elapsed between drug administration and sampling. Samples for one patient, withdrawn 24 min after dosing, had ketorolac concentrations below the quantification limit. The ratios in the remaining patients were all low and showed a tendency to increase with time. The mean ratio was 0.116 with a range of 0.04 in 2 patients, at 43 min and 1 h 6 min, to 0.25 at 6 h 34 min.


The Entamoeba histolytica methylated LINE-binding protein EhMLBP provides protection against heat shock.
Adaptation to environmental stress is a key process that allows the unicellular parasite Entamoeba histolytica to survive in its human host. We previously characterized EhMLBP as an essential protein for the growth and the virulence of the parasite. EhMLBP binds to methylated repetitive DNA, and is one of the core proteins of the parasite's epigenetic machinery. Here, we show that EhMLBP and heat shock proteins have common properties. EhMLBP is induced by heat shock and its expression is regulated by a heat shock element binding site that is located in its 5' non-coding region. Following heat shock, the perinuclear localization of EhMLBP in control trophozoites is replaced by an even distribution within the nucleus alongside with an enhanced recruitment of EhMLBP to the reverse transcriptase of a long interspersed nucleotide element (LINE) DNA. Constitutive overexpression of EhMLBP protects trophozoites against heat shock and reduces protein aggregation. This protective function is lost in trophozoites that overexpress a mutated form of EhMLBP which is devoid of its heat shock domain. To the best of our knowledge, this is the first report of a methyl DNA-binding protein that plays a protective role against heat shock.


PK study of octreotide based on LC-MS/MS combining protein precipitation and impurity extraction technique.
To establish a robust methodology for quantitative analysis of therapeutic peptide in biological samples. Octreotide was chosen as a model therapeutic peptide, and oxidized-octreotide was synthesized as internal standard. Protein precipitation combining liquid-liquid extraction technique was adopted to enhance the recovery and reduce the endogenous interferences effectively. A LC-MS/MS method for the quantification of octreotide in plasma has been optimized and validated according to FDA guidelines. Linearity, selectivity, accuracy, precision, stability, matrix effect and recovery were within bioanalytical method validation acceptance criteria as FDA guidelines. The methodology was then successfully applied into the studies for octreotide. This robust methodology would be useful for the PK studies for octreotide and other therapeutic peptides.


Prognostic significance of microvessel density determined by an anti-CD105/endoglin monoclonal antibody in astrocytic tumors: comparison with an anti-CD31 monoclonal antibody.
There are conflicting reports as to whether the degree of angiogenesis as measured by microvessel density (MVD) has a prognostic value in astrocytic tumors. This may be due to the use of different antibodies against endothelial cells to highlight microvessels. It has been reported that unlike pan-endothelial antibodies, such as CD31, anti-CD105 antibodies preferentially react with endothelial cells in angiogenic tissues. To clarify the validity of anti-CD105 antibody in the evaluation of angiogenesis, we assessed MVD using an anti-CD105 monoclonal antibody (mAb) (CD105-MVD) and an anti-CD31 mAb (CD31-MVD) in a series of 50 astrocytic tumors, and correlated MVD with expression of the key angiogenic factor vascular endothelial growth factor (VEGF) and prognosis. The mean CD31-MVD and CD105-MVD was 36.7 and 24.8 for low-grade astrocytoma (LGA), 48.0 and 42.7 for anaplastic astrocytoma, 55.3 and 51.9 for glioblastoma multiforme (GBM), respectively. CD105-MVD was more closely correlated with VEGF expression than CD31-MVD. Patients with LGA and GBM showing higher CD105-MVD had a significantly shorter mean survival time (MST) than those with lower CD105-MVD tumors (P = 0.0381 and 0.0131, respectively). Whereas the MST of patients with higher CD31-MVD tumors seemed to be shorter than that of lower CD31-MVD patients within each tumor grade, the differences were not statistically significant. These findings suggest that anti-CD105 mAb may be a better marker than anti-CD31 mAb in evaluation of angiogenesis and prediction of prognosis in astrocytic tumors.