Rate of change in functional limitations for patients with rheumatoid arthritis: effects of sex, age, and duration of illness.
To estimate the rate of change in functional limitations for patients with rheumatoid arthritis (RA) as a function of age, duration of illness, and sex. Patients with RA (n = 700) aged 21-65 years in 1988 were interviewed yearly for 6 years in The National Rheumatoid Arthritis Study. Functional limitations scores based on a Rasch measurement model of 20 Health Assessment Questionnaire items were analyzed in mixed-effects models to estimate the rate of change in functional ability as a function of age, duration of illness, sex, and interactions. Models for both patient age and duration of illness significantly predicted limitations in functional ability for men and women. The model for age included a significant cubic effect; the model for duration of illness included a significant linear effect only. Sex was significant in both models and no interactions were significant in either model. The AIC index of fit, an indicator of the information value of the model, favored the model for duration of illness over the model for age. While both models showed higher levels of functional limitations in women than men, the rate of change for women was similar to men. Limitation in functional ability in RA progressed in a linear manner with duration of illness and progressed at the same rate for both men and women, but functional limitations were greater for women.


20 pack-year smoking history as strongest smoking metric predictive of HPV-positive oropharyngeal cancer outcomes.
While smoking is associated with worse outcomes in HPV-positive oropharyngeal squamous cell carcinoma (OPSCC), the magnitude of this association is unclear given the heterogenous smoking definitions and outcomes. Our objective was to investigate the association between smoking, survival, and recurrence in HPV-related OPSCC using multiple smoking metrics reported in the literature. This was a retrospective cohort study of 375 adults with p16+ OPSCC undergoing surgical resection (n = 272) or definitive chemoradiation (n = 103) at a tertiary academic institution from 2006 to 2017. The primary outcome was overall survival (OS). Secondary outcomes included disease-free survival (DFS), disease-specific survival (DSS), and recurrence. We used multiple smoking metrics commonly cited in previous studies, including ever versus never smokers, current versus former/never smokers, ≤10 versus >10 pack-year, ≤20 versus >20 pack-year, and continuous pack-year. There were 375 patients, median age 58 years, with 326 (87%) males, and median follow-up of 52 months. Of all smoking metrics, >20 pack-year history was the strongest predictor of both OS (HR 2.24, 95% CI: 1.19-4.20) and DFS (HR 1.67, 95% CI: 1.04-2.66) on univariable and multivariable analysis after adjusting for age, overall stage, and comorbidities. Patients with >20 pack-year smoking history were also more likely to have recurrence (HR 1.59, 95% CI: 0.95-2.67) after adjusting for overall stage. Heavier smoking >20 pack-years was the strongest smoking metric associated with 2-times worse survival and recurrence. Our findings suggest that >20 pack-year smoking history may be a more useful cutoff for risk stratification models but requires further validation.


Rapid response team activation prior to in-hospital cardiac arrest: Areas for improvements based on a national cohort study.
Rapid response teams (RRTs) are designed to improve the "chain of prevention" of in-hospital cardiac arrest (IHCA). We studied the 30-day survival of patients reviewed by RRTs within 24 hours prior to IHCA, as compared to patients not reviewed by RRTs. A nationwide cohort study based on the Swedish Registry of Cardiopulmonary Resuscitation, between January 1st, 2014, and December 31st, 2021. An explorative, hypothesis-generating additional in-depth data collection from medical records was performed in a small subgroup of general ward patients reviewed by RRTs. In all, 12,915 IHCA patients were included. RRT-reviewed patients (n = 2,058) had a lower unadjusted 30-day survival (25% vs 33%, p < 0.001), a propensity score based Odds ratio for 30-day survival of 0.92 (95% Confidence interval 0.90-0.94, p < 0.001) and were more likely to have a respiratory cause of IHCA (22% vs 15%, p < 0.001). In the subgroup (n = 82), respiratory distress was the most common RRT trigger, and 24% of the RRT reviews were delayed. Patient transfer to a higher level of care was associated with a higher 30-day survival rate (20% vs 2%, p < 0.001). IHCA preceded by RRT review is associated with a lower 30-day survival rate and a greater likelihood of a respiratory cause of cardiac arrest. In the small explorative subgroup, respiratory distress was the most common RRT trigger and delayed RRT activation was frequent. Early detection of respiratory abnormalities and timely interventions may have a potential to improve outcomes in RRT-reviewed patients and prevent further progress into IHCA.


Botulinum toxin A injection into the entopeduncular nucleus improves dynamic locomotory parameters in hemiparkinsonian rats.
Parkinson's disease is associated with hyperactivity of the subthalamic nucleus (STN), contributing to motor and gait disturbances. Although deep brain stimulation of the STN alleviates certain motor dysfunction, its specific effect on gait abnormalities remains controversial. This study investigated the long-term changes in locomotion following direct infusions of botulinum toxin-A into the globus pallidus internal segment (GPi) to suppress the flow of information from the STN to the GPi in a hemiparkinsonian rat model. Static and dynamic gait parameters were quantified using a CatWalk apparatus. Interestingly, botulinum toxin-A at 0.5 ng significantly reduced only the dynamic gait parameters of hemiparkinsonian rats at 1 week and 1 month post-infusion, while static gait parameters did not change. This study offers new insights into the complexity of basal ganglia in locomotor control and shows the potential of central infusion of botulinum toxin-A as a novel intervention in the study of experimental hemiparkinson's disease.


Keratocyte Differentiation Is Regulated by NF-κB and TGFβ Signaling Crosstalk.
Interleukin-1 (IL-1) and transforming growth factor-beta (TGFβ) are important cytokines involved in corneal wound healing. Here, we studied the effect of these cytokines on corneal stromal cell (keratocyte) differentiation. IL-1β treatment resulted in reduced keratocyte phenotype, as evident by morphological changes and decreased expression of keratocyte markers, including keratocan, lumican, ALDH3A1, and CD34. TGFβ1 treatment induced keratocyte differentiation towards the myofibroblast phenotype. This was inhibited by simultaneous treatment with IL-1β, as seen by inhibition of α-SMA expression, morphological changes, and reduced contractibility. We found that the mechanism of crosstalk between IL-1β and TGFβ1 occurred via regulation of the NF-κB signaling pathway, since the IL-1β induced inhibition of TGFβ1 stimulated keratocyte-myofibroblast differentiation was abolished by a specific NF-κB inhibitor, TPCA-1. We further found that Smad7 participated in the downstream signaling. Smad7 expression level was negatively regulated by IL-1β and positively regulated by TGFβ1. TPCA-1 treatment led to an overall upregulation of Smad7 at mRNA and protein level, suggesting that NF-κB signaling downregulates Smad7 expression levels in keratocytes. All in all, we propose that regulation of cell differentiation from keratocyte to fibroblast, and eventually myofibroblast, is closely related to the opposing effects of IL-1β and TGFβ1, and that the mechanism of this is governed by the crosstalk of NF-κB signaling.


Determining when quality improvement initiatives should be considered research: proposed criteria and potential implications.
Quality improvement initiatives have become a major force in shaping health care. These initiatives are diverse but share a common goal of generating knowledge that will guide improvements in health care. Although quality improvement occupies an uncertain territory between clinical care and research, this difference is extremely important. If a quality improvement initiative is research, federal regulations require a variety of procedures that must be followed to protect the human subjects involved. Conversely, if the same initiative is not research, the regulations outlining these procedures are not applicable. This article proposes 2 criteria to determine whether a quality improvement initiative should be reviewed as research. Such an initiative should be reviewed and regulated as research if (1) the majority of patients involved are not expected to benefit directly from the knowledge to be gained or (2) if additional risks or burdens are imposed to make the results generalizable. Implementation of these criteria offers the possibility that the rights and interests of those who participate in quality improvement initiatives will be protected.


Incorporation of highly purified melittin into phosphatidylcholine bilayer vesicles.
Melittin free of phospholipase A2 was prepared. In the absence of salt this highly pure protein starts to aggregate in solution at a protein concentration of Cp greater than 10(-3) M. In high salt solution (2 M) aggregation starts at Cp greater than 10(-6) M. This was determined from the blue shift of the intrinsic fluorescence of the protein. Reinvestigation of the quenching behaviour clearly shows that self-aggregation cannot be deduced from quenching experiments using nitrate or 2,2,6,6-tetramethylpiperidine-1-oxyl as quencher. The incorporation of melittin into phosphatidylcholine bilayer vesicles was studied by fluorescence quenching and by energy-transfer experiments using 2- and 6-anthroyloxypalmitic acid as acceptor and peptide tryptophan as donor. Incorporation of melittin into small unilamellar vesicles was found to be reduced below the lipid phase transition temperature, Tt, whereas it incorporates and distributes more randomly above Tt. Cooling the temperature below Tt after incubation at T greater than Tt leads to a deeper incorporation of the peptide into the lipid bilayer due to electrostatic interaction between the lipid phosphate groups and the positively charged amino acids. This stabilizing effect is lost above Tt and melittin is extruded to the polar phase. Quenching experiments support this finding. EPR measurements clearly demonstrate that even in the presence of high amounts of melittin up to 10 mol% with respect to the lipid broadening of the phase transition curves was only observed with fatty acid spin labels, where the doxyl group is localized near the bilayer surface. The order degree of the inner part of the bilayer remains almost unchanged even in the presence of high melittin content.


Human antibody response during sepsis against targets expressed by methicillin resistant Staphylococcus aureus.
The identification of target structures is a prerequisite for the development of new treatment options, like antibody based therapy, against methicillin resistant Staphylococcus aureus (MRSA). In this study we identified immunodominant structures which were expressed in vivo during sepsis caused by MRSA. Using human sera we compared the immune response of humans with MRSA sepsis with the immune response of normal individuals and asymptomatically colonized individuals. We identified and characterized four staphylococcal specific antigenic structures. One target is a staphylococcal protein of 29 kDa that exhibited 29% identity to secreted protein SceA precursor of Staphylococcus carnosus. The putative function of this protein, which was designated IsaA (immunodominant staphylococcal antigen), is unknown. The second target is an immunodominant protein of 17 kDa that showed no homology to any currently known protein. This immunodominant protein was designated IsaB. The third and fourth antigens are both immunodominant proteins of 10 kDa. One of these proteins showed 100% identity to major cold shock protein CspA of S. aureus and the other protein was identified as the phosphocarrier protein Hpr of S. aureus. The identified immunodominant proteins may serve as potential targets for the development of antibody based therapy against MRSA.


Accumulation of rare earth elements in corn after agricultural application.
Using both pot and plot experiments, the dose-dependent accumulation of rare earth elements (REs) in corn (Zea mays L.) after application of an agricultural REs mixture was measured. In the pot experiment, the dose-dependent accumulation of REs in corn root and stem was observed, but it could not be detected in corn leaf under the dosage of 20 mg REs kg(-1) soil (oven-dry mass). The non-observed effect concentration (NOEC) for accumulation of REs in corn seedling with the pot experiment was 1.0 mg REs kg(-1). In the plot experiment, the dose-dependent accumulation was observed at an early stage after application of REs and the NOEC value of 32 mg REs m(-2) was obtained. At harvest, no dose-dependent accumulation of REs was observed in any part of the corn. These results can be confirmed by the fingerprinting analysis based on the differences between La to RE ratios in the REs mixture and in pot or plot soil. We observed that the plant shows no preference on individual RE and the results of fingerprinting indicated clearly the incorporation of exogenous REs in plant tissues, in a similar manner as that observed in the dose-dependent distribution of RE concentrations. The results indicated also a translocation process of REs from plant root to leaf when applied to soil or from leaf to root when applied to leaf. A homeostatic regulation mechanism for excessive uptake of REs in plants is suggested to regulate the concentrations of REs in the plant.


Desensitized hologram interferometry.
The possibility of using multiple-wavelength holography to obtain desensitized hologram interferometry is being investigated analytically and experimentally. A simple analysis has been performed, and its conclusions are supported by experimental results. These results indicate that the new techniques will extend by several orders of magnitude the range of displacements amenable to measurement to hologram interferometry.


Metabolic alterations in cerebrospinal fluid from double hemorrhage model of dogs.
Even though cerebral vasospasm after subarachnoid hemorrhage (SAH) causes cerebral ischemia or infarction, the metabolic alterations in cerebrospinal fluids (CSF) after SAH have not been studied. This study was undertaken to measure the levels of glucose, lactate, pyruvate and glutamate in CSF from double hemorrhage dog models. Thirty-two mongrel dogs of either sex, weighing 18-24 kg, underwent double hemorrhage by percutaneous needle puncture of the cistema magna and injection of autologous blood on day 0 and day 2. The dogs were then sacrificed on day 3, 5 and 7, after collecting CSF. In another study, the dogs were treated with mitogen-activated protein kinase (MAPK) inhibitors PD98059 and U0126, and caspase-2 and caspase-3 inhibitors from day 3 to day 6 after initial blood injection. CSF was collected on day 7 before dogs were sacrificed. The concentration of glucose, lactate, pyruvate and glutamate in CSF was measured by photometrical method. Compared with CSF collected on day 0, glucose was decreased on days 5-7, lactate was increased on days 2-7, pyruvate was increased on days 2-7, and glutamate was increased on days 3-7 (p < 0.05). In the groups treated with MAPK or caspase inhibitors, most of the metabolic alterations remained unchanged as compared with CSF from untreated dogs. Clinically, caspase inhibitors-2 and -3, and MAPK inhibitor U0126 all failed to prevent vasospasm. MAPK inhibitor PD98059 partially prevented vasospasm. Our data demonstrated a metabolic alteration of glucose, glutamate, lactate and pyruvate in CSF during cerebral vasospasm. This metabolic change in consistent with the time course of cerebral vasospasm. This study suggests that brain energy metabolites and excitative amino acids are altered during cerebral vasospasm.


Current status of cyhalofop-butyl and metamifop resistance and diversity of the ACCase gene mutations in Chinese sprangletop (Leptochloa chinensis) from China.
Leptochloa chinensis populations in China have evolved widespread resistance to acetyl coenzyme A carboxylase (ACCase)-inhibiting herbicides cyhalofop-butyl (CyB) and metamifop (Met). 124 L. chinensis populations, randomly collected from rice fields in Jiangsu Province, were surveyed for CyB and Met resistance status, and all potential ACCase gene resistance-conferring mutations and effective pre-emergence herbicides for its control were investigated. Single-dose tests confirmed that 82 (66.1%) and 70 (56.4%) populations evolved resistance to CyB and Met, respectively. ACCase sequencing revealed that 56.4% of the populations contain plants with diverse target-site ACCase mutations (Ile1781Leu, Trp1999Cys, Trp2027Cys, Trp2027Ser, Ile2041Asn, Gly2096Ala, and in particular, a Leu1818Phe mutation). Notably, the Leu1818Phe mutation had been detected in 8 resistant populations, indicating this mutation was prone to occur in L. chinensis. Additionally, 9.7% of the populations may have single metabolic resistance to CyB, as these populations was susceptible to Met, and no any ACCase mutations were found. Moreover, the resistant populations with different ACCase mutations showed 6.5 to 33.6-fold resistance to CyB, and 4.4 to 82.6-fold resistance to Met. Importantly, five pre-emergence herbicides, including pretilachlor, pendimethalin, clomazone, pyraclonil, and mefenacet, all exhibited good control effect on resistant L. chinensis populations. This work confirmed the prevalence and distribution of CyB and Met resistance in L. chinensis. Target-site ACCase mutations made a major contribution to CyB and Met resistance. Pre-emergence herbicides could be valuable tools for management of resistant L. chinensis populations.


Considerations for management of interstitial ectopic pregnancies: two case reports.
Conventional treatment of interstitial pregnancies includes systemic methotrexate, direct methotrexate injection, wedge resection, or hysterectomy. We present two cases of interstitial pregnancies that were successfully managed by different minimally invasive surgical techniques. We also report the novel use of hysteroscopic urologic stone retrieval forceps in the transvaginal removal of persistent products of conception after systemic methotrexate for an interstitial pregnancy. Case 1 was a 28-year-old gravida 1 white woman at 8 weeks gestation; she was diagnosed with a left interstitial pregnancy. After laparoscopic confirmation of the interstitial pregnancy, successful ultrasound-guided suction dilation and curettage was performed. Case 2 was a 33-year-old gravida 3 para 1021 (one term pregnancy, no preterm pregnancies, one ectopic pregnancy and one spontaneous miscarriage, and one living child) Hispanic woman with persistent products of conception after systemic methotrexate for a left interstitial pregnancy. She underwent hysteroscopic-guided removal of the persistent products of conception, which was possible due to novel use of urologic stone retrieval forceps. Successful minimally invasive treatment of interstitial pregnancies may be possible in certain cases. Collaboration between different specialties continues to be important for improving minimally invasive options.


Transcriptional gene expression profiles of HGF/SF-met signaling pathway in colorectal carcinoma.
To explore the transcriptional gene expression profiles of HGF/SF-met signaling pathway in colorectal carcinoma to understand mechanisms of the signaling pathway at so gene level. Total RNA was isolated from human colorectal carcinoma cell line LoVo treated with HGF/SF (80 ng/L) for 48 h. Fluorescent probes were prepared from RNA labeled with cy3-dUTP for the control groups and with cy5-dUTP for the HGF/SF-treated groups through reverse-transcription. The probes were mixed and hybridized on the microarray at 60 degrees for 15-20 h, then the microarray was scanned by laser scanner (GenePix 4000B). The intensity of each spot and ratios of Cy5/Cy3 were analyzed and finally the differentially expressed genes were selected by GenePix Pro 3.0 software. 6 differential expression genes(3 up-regulated genes and 3 down-regulated genes) were selected randomly and analyzed by beta-actin semi-quantitative RT-PCR. The fluorescent intensities of built-in negative control spots were less than 200, and the fluorescent intensities of positive control spots were more than 5000. Of the 4004 human genes analyzed by microarray, 129 genes (holding 3.22 % of the investigated genes) revealed differential expression in HGF/SF-treated groups compared with the control groups, of which 61 genes were up-regulated (holding 1.52 % of the investigated genes) and 68 genes were down-regulated (holding 1.70 % of the investigated genes), which supplied abundant information about target genes of HGF/SF-met signaling. HGF/SF-met signaling may up-regulate oncogenes, signal transduction genes, apoptosis-related genes, metastasis related genes, and down-regulate a number of genes. The complexity of HGF/SF-met signaling to control the gene expression is revealed as a whole by the gene chip technology.


An administrative claims model for profiling hospital 30-day mortality rates for pneumonia patients.
Outcome measures for patients hospitalized with pneumonia may complement process measures in characterizing quality of care. We sought to develop and validate a hierarchical regression model using Medicare claims data that produces hospital-level, risk-standardized 30-day mortality rates useful for public reporting for patients hospitalized with pneumonia. Retrospective study of fee-for-service Medicare beneficiaries age 66 years and older with a principal discharge diagnosis of pneumonia. Candidate risk-adjustment variables included patient demographics, administrative diagnosis codes from the index hospitalization, and all inpatient and outpatient encounters from the year before admission. The model derivation cohort included 224,608 pneumonia cases admitted to 4,664 hospitals in 2000, and validation cohorts included cases from each of years 1998-2003. We compared model-derived state-level standardized mortality estimates with medical record-derived state-level standardized mortality estimates using data from the Medicare National Pneumonia Project on 50,858 patients hospitalized from 1998-2001. The final model included 31 variables and had an area under the Receiver Operating Characteristic curve of 0.72. In each administrative claims validation cohort, model fit was similar to the derivation cohort. The distribution of standardized mortality rates among hospitals ranged from 13.0% to 23.7%, with 25(th), 50(th), and 75(th) percentiles of 16.5%, 17.4%, and 18.3%, respectively. Comparing model-derived risk-standardized state mortality rates with medical record-derived estimates, the correlation coefficient was 0.86 (Standard Error = 0.032). An administrative claims-based model for profiling hospitals for pneumonia mortality performs consistently over several years and produces hospital estimates close to those using a medical record model.


Hymenoptera Venom Allergy. A closer collaboration is needed between allergists and emergency physicians.
Background. Hymenoptera stings are sometimes fatal in venom-allergic patients. Fatalities mostly occur in previously stung subjects, especially those with a history of systemic reactions, and could be avoided if patients were properly informed of the existence of a prevention strategy for insect stings, referred to an allergy follow-up and prescribed auto-injectable epinephrine and/or venom-specific immunotherapy (VIT). We sought to assess knowledge and awareness of Hymenoptera Venom Allergy (HVA) in a small sample of Emergency Physicians in our geographic area. Methods. An eight-point questionnaire on HVA was administered to Emergency Department physicians working in the six largest ED in Naples. Results. Twenty-seven physicians completed the questionnaire. Twenty/27 (74%) were unaware of the classification of Hymenoptera sting reactions, 11/27 (41%) were unaware of the existence of prevention strategies such as VIT, 18/27 (67%) did not refer HVA patients to a specialist follow up. One/27 (4%) prescribed auto-injectable epinephrine and 100% wish better information on the topic. Conclusions. In our survey we found a number of ED physicians whose knowledge of HVA, beyond the emergency treatment, is not satisfactory. A closer collaboration among ED physicians and allergists is urgently needed.


Phase I/IIa study of intratumoral/intracerebral or intravenous/intracerebral administration of Parvovirus H-1 (ParvOryx) in patients with progressive primary or recurrent glioblastoma multiforme: ParvOryx01 protocol.
The treatment of patients with malignant brain tumors remains a major oncological problem. The median survival of patients with glioblastoma multiforme (GBM), the most malignant type, is only 15 months after initial diagnosis and even less after tumor recurrence. Improvements of standard treatment including surgery and radio-chemotherapy have not lead to major improvements. Therefore, alternative therapeutics such as oncolytic viruses that specifically target and destroy cancer cells are under investigation. Preclinical data of oncolytic parvovirus H-1 (H-1PV) infection of glioma cells demonstrated strong cytotoxic and oncosuppressing effects, leading to a phase I/IIa trial of H-1PV in patients with recurrent GBM (ParvOryx01). ParvOryx01 is the first trial with a replication competent oncolytic virus in Germany. ParvOryx01 is an open, non-controlled, two groups, intra-group dose escalation, single center, phase I/IIa trial. 18 patients with recurrent GBM will be treated in 2 groups of 9 patients each. Treatment group 1 will first receive H-1PV by intratumoral injection and second by administration into the walls of the tumor cavity during tumor resection. In treatment group 2 the virus will initially be injected intravenously and afterwards, identical to group 1, into the surrounding brain tissue during tumor removal. Main eligibility criteria are: age of 18 years, unifocal recurrent GBM, amenable to complete or subtotal resection. Dose escalation will be based on the Continual Reassessment Method. The primary objective of the trial is local and systemic safety and tolerability and to determine the maximum tolerated dose (MTD). Secondary objectives are proof of concept (PoC) and Progression-free Survival (PFS) up to 6 months. This is the first trial with H-1PV in patients with recurrent GBM. The risks for the participants appear well predictable and justified. Furthermore, ParvOryx01 will be the first assessment of combined intratumoral and intravenous application of an oncolytic virus. Due to its study design the trial will not only generate data on the local effect of H-1PV but it will also investigate the penetration of H-1PV into the tumor after systemic delivery and obtain safety data from systemic delivery possibly supporting clinical trials with H-1PV in other, non-CNS malignancies. ClinicalTrials.gov Identifier: NCT01301430.


Philip Morris involvement in the development of an air quality laboratory in El Salvador.
The tobacco industry has organised research institutions to generate misleading data on indoor air quality, including second-hand smoke exposure and health effects. To describe tobacco industry involvement in the organisation and financial support of an air quality research laboratory in El Salvador. Tobacco industry documents on the internet were systematically searched from August 2007 to February 2008 for air quality studies undertaken in El Salvador, and laboratory personnel were interviewed. Philip Morris sought to establish a network of air quality laboratories throughout Latin America. In El Salvador, in 1997, through Tabacalera de El Salvador (a subsidiary of Philip Morris) and the Salvadoran Foundation for Economic Development (FUSADES), the industry organised an air quality research laboratory. FUSADES was part of the industry's Latin American Scientific Network, which consisted of doctors hired as consultants who would send air samples from their research to FUSADES. Philip Morris Scientific Affairs personnel hired LabStat, a Canadian-based laboratory, to provide technical assistance to FUSADES (train and assist the laboratory in air quality measurements). In addition, the Washington-based HMS Group successfully implemented a plan to upgrade the laboratory and obtain international certifications. HMS Group also assisted in searching for sustainable funding for FUSADES, including seeking funds from international aid for Hurricane Mitch. Air quality studies that have used the FUSADES laboratory should be carefully interpreted, given the support that this laboratory received from Philip Morris.


Antiangiogenic tyrosine kinase inhibitors in colorectal cancer: is there a path to making them more effective?
Antiangiogenic therapy has a proven survival benefit in metastatic colorectal cancer. Inhibition of the VEGF pathway using a variety of extracellular antibody approaches has clear benefit in combination with chemotherapy, while intracellular blockade using tyrosine kinase inhibitors (TKIs) such as sorafenib and regorafenib has had more limited success. Pharmacodynamic modeling using modalities such as DCE-MRI indicates potent antiangiogenic effects of these TKIs, yet numerous combination therapies, primarily with chemotherapy, have failed to demonstrate an additive benefit. The sole comparative study of a single agent TKI against placebo showed a survival benefit of regorafenib in patients with advanced, refractory disease. Preclinical data demonstrate synergy between antiantiogenic TKIs and targeted interventions including autophagy inhibition, and together with a renewed effort to define markers of susceptibility, such combinations may be a way to improve the limited efficacy of this once-promising drug class.


Quantitative and qualitative analysis of antimicrobial usage patterns in 180 selected farrow-to-finish pig farms from nine European countries based on single batch and purchase data.
Farm-level quantification of antimicrobial usage (AMU) in pig farms. In a cross-sectional study, AMU data on group treatments administered to a single batch of fattening pigs from birth to slaughter (group treatment data) and antimicrobials purchased during 1 year (purchase data) were collected at 180 pig farms in nine European countries. AMU was quantified using treatment incidence (TI) based on defined (DDDvet) and used (UDDvet) daily doses and defined (DCDvet) and used (UCDvet) course doses. The majority of antimicrobial group treatments were administered to weaners (69.5% of total TIDDDvet) followed by sucklers (22.5% of total TIDDDvet). AMU varied considerably between farms with a median TIDDDvet of 9.2 and 7.1 for a standardized rearing period of 200 days based on group treatment and purchase data, respectively. In general, UDDvet and UCDvet were higher than DDDvet and DCDvet, respectively, suggesting that either the defined doses were set too low or that group treatments were often dosed too high and/or administered for too long. Extended-spectrum penicillins (31.2%) and polymyxins (24.7%) were the active substances most often used in group treatments, with the majority administered through feed or water (82%). Higher AMU at a young age was associated with higher use in older pigs. Collecting farm-level AMU data of good quality is challenging and results differ based on how data are collected (group treatment data versus purchase data) and reported (defined versus used daily and course doses).


Abdominal fat depots and their association with insulin resistance in patients with type 2 diabetes.
Asian-Indians show thin fat phenotype, characterized by predominantly central deposition of excess fat. The roles of abdominal subcutaneous fat (SAT), intra-peritoneal adipose tissue, and fat depots surrounding the vital organs (IPAT-SV) and liver fat in insulin resistance (IR), type-2 diabetes (T2D) and metabolic syndrome (MetS) in this population are sparsely investigated. Assessment of liver fat, SAT and IPAT-SV by MRI in subjects with T2D and MetS; and to investigate its correlation with IR, specifically according to different quartiles of HOMA-IR. Eighty T2D and the equal number of age sex-matched normal glucose tolerant controls participated in this study. Abdominal SAT, IPAT-SV and liver fat were measured using MRI. IR was estimated by the Homeostatic Model Assessment for Insulin Resistance (HOMA-IR). T2D and MetS subjects have higher quantity liver fat and IPAT-SV fat than controls (P = 9 x 10-4 and 4 x 10-4 for T2D and 10-4 and 9 x 10-3 for MetS subjects respectively). MetS subjects also have higher SAT fat mass (P = 0.012), but not the BMI adjusted SAT fat mass (P = 0.48). Higher quartiles of HOMA-IR were associated with higher BMI, W:H ratio, waist circumference, and higher liver fat mass (ANOVA Test P = 0.020, 0.030, 2 x 10-6 and 3 x 10-3 respectively with F-values 3.35, 3.04, 8.82, 4.47 respectively). In T2D and MetS subjects, HOMA-IR showed a moderately strong correlation with liver fat (r = 0.467, P < 3 x 10-5 and r = 0.493, P < 10-7), but not with SAT fat and IPAT-SV. However, in MetS subjects IPAT-SV fat mass showed borderline correlation with IR (r = 0.241, P < 0.05), but not with the BMI adjusted IPAT-SV fat mass (r = 0.13, P = 0.26). In non-T2D and non-MetS subjects, no such correlation was seen. On analyzing the correlation between the three abdominal adipose compartment fat masses and IR according to its severity, the correlation with liver fat mass becomes stronger with increasing quartiles of HOMA-IR, and the strongest correlation is seen in the highest quartile (r = 0.59, P < 10-3). On the other hand, SAT fat mass tended to show an inverse relation with IR with borderline negative correlation in the highest quartile (r = -0.284, P < 0.05). IPAT-SV fat mass did not show any statistically significant correlation with HOMA-IR, but in the highest quartile it showed borderline, but statistically insignificant positive correlation (P = 0.07). In individuals suffering from T2D and MetS, IR shows a trend towards positive and borderline negative correlation with liver fat and SAT fat masses respectively. The positive trend with liver fat tends to become stronger with increasing quartile of IR. Therefore, these findings support the theory that possibly exhaustion of protective compartment's capacity to store excess fat results in its pathological deposition in liver as ectopic fat.


Measurement of body composition in cats using computed tomography and dual energy X-ray absorptiometry.
Dual energy X-ray absorptiometry (DEXA) is a reference method for assessing body composition but is seldom `accessible in veterinary settings. Computed tomography (CT) can provide similar body composition estimates and we propose that it can be used in body composition studies in animals. We compared CT and DEXA data from 73 healthy adult neutered domestic cats. Three approaches for measuring adipose tissue percentage from full-body CT scans were explored. By examining the frequency distribution of voxels by Hounsfield unit (HU) value, it is possible to calculate a fat index (Fat%) that is in close agreement with the fat percentages obtained from DEXA scans. Fat% values obtained by the best of the methods had a mean difference of 0.96% (95% confidence interval 0.33-1.59%) from the DEXA results. Fat% obtained by the other two methods were characterized by good correlation but poor agreement and in one of the methods, the difference between the values from the two modalities was proportional to their mean. By using CT, it is possible to obtain body composition estimates that are in close agreement with those available using DEXA. While the significance of individual Fat% measurements obtained from CT can be difficult to interpret and to compare between centers, CT can contribute to research studies concerned either with nutrition or with obesity-related disorders.


Coevolution of killer cell Ig-like receptors with HLA-C to become the major variable regulators of human NK cells.
Interactions between HLA class I and killer cell Ig-like receptors (KIRs) diversify human NK cell responses. Dominant KIR ligands are the C1 and C2 epitopes of MHC-C, a young locus restricted to humans and great apes. C1- and C1-specific KIRs evolved first, being present in orangutan and functionally like their human counterparts. Orangutans lack C2 and C2-specific KIRs, but have a unique C1+C2-specific KIR that binds equally to C1 and C2. A receptor with this specificity likely provided the mechanism by which C2-KIR interaction evolved from C1-KIR while avoiding a nonfunctional intermediate, that is, either orphan receptor or ligand. Orangutan inhibitory MHC-C-reactive KIRs pair with activating receptors of identical avidity and specificity, contrasting with the selective attenuation of human activating KIRs. The orangutan C1-specific KIR reacts or cross-reacts with all four polymorphic epitopes (C1, C2, Bw4, and A3/11) recognized by human KIRs, revealing their structural commonality. Saturation mutagenesis at specificity-determining position 44 demonstrates that KIRs are inherently restricted to binding just these four epitopes, either individually or in combination. This restriction frees most HLA-A and HLA-B variants to be dedicated TCR ligands, not subject to conflicting pressures from the NK cell and T cell arms of the immune response.


Ultrasonography of the lower extremity.
Ultrasonography is a useful imaging tool for various soft tissue and joint pathologies affecting the lower extremity. This article reviews the normal sonographic appearance of muscles, tendons, ligaments, nerves, bone, and cartilage. The ultrasound imaging appearance of various pathologic conditions affecting the hip, thigh, knee, lower leg, ankle,and foot are illustrated. The advantages of ultrasonography are highlighted.


Lasofoxifene: Evidence of its therapeutic value in osteoporosis.
Osteoporosis is a skeletal disorder characterized by compromised bone strength and increased risk of fracture. It is a common disorder in elderly subjects and represents a major public health problem, affecting up to 40% postmenopausal women and 15% of men. Among the several therapeutical interventions, hormone replacement therapy (HRT) was traditionally seen as the gold standard for preventing osteoporotic fractures in postmenopausal women, as well as for the management of menopausal symptoms. However HRT, especially if administered long-term, may lead to an increased risk of breast and, when unopposed by progestins, endometrial cancers. Alternative therapies include bisphosphonates and raloxifene, a selective estrogen receptor modulator (SERM). While the former have been associated with suboptimal adherence, the latter was considerably less potent than estrogen and its effect in the prevention of nonvertebral fractures remain uncertain. The purpose of this article is to review the clinical trials of lasofoxifene, a new SERM for the treatment of postmenopausal osteoporosis. The medical literature was reviewed for appropriate articles containing the terms "lasofoxifene" and SERMs". There are three (phase II or phase III) clinical trials that clearly demonstrate efficacy and safety of this new SERM in the suppression of bone loss and the prevention of vertebral and nonvertebral fractures. Moreover, lasofoxifene treatment also reduced breast cancer risk and the occurrence of vaginal atrophy. With its increased potency and efficacy on the prevention of nonvertebral fractures lasofoxifene may be an alternative and cost-effective therapy for osteoporosis in postmenopausal women.


Is it a vicious circle and for whom? The reciprocal association between rumination and somatic symptoms and moderation by stress: A daily diary study among Chinese college students.
College students who experienced somatic symptoms during the COVID-19 pandemic may engage in rumination, but their bidirectional nature remains underexplored. Symptom perception theory suggests a reciprocal relationship between rumination and somatic symptoms, and the multiple-stressor perspective and the perseverative cognition hypothesis assume that the reciprocal association might be exacerbated by high stress. In this study, we examined temporal associations between rumination and somatic symptoms and variations by patterns of stress related to COVID-19 and daily hassles. A total of 582 Chinese college students provided daily reports on rumination, somatic symptoms, COVID-related stress, and daily hassles for seven consecutive days in November 2020. A cross-lagged panel model showed a positive reciprocal association between rumination and somatic symptoms. Greater rumination predicted more next-day somatic symptoms, and more somatic symptoms increased next-day rumination. Dual trajectory analysis identified four stress patterns of COVID-related stress and daily hassles (i.e. low-low, low-high, high-low, and high-high), and multi-group analysis found the reciprocal association only presented in the high-high group. Our findings indicate a vicious circle between rumination and somatic symptoms that is dependent on heterogeneous stress patterns. Attention should be paid to the high-risk group with both high levels of COVID-related stress and daily hassles.


Quantification of temporal lobe resections: a new approach.
Extent of resection in temporal lobectomy has been traditionally based on intraoperative linear measurements. The complex geometrical contour of the temporal lobe limits the precision and accuracy of such measurements, and it is often difficult to verify the extent of actual resection after surgery. The authors propose a new method of evaluating extent of resection based on a 20-compartment model of the temporal lobe. The temporal lobe is divided into five anteroposterior coronal sections in relation to the anterior and posterior borders of the mesencephalon. Each section is then divided into superior lateral, inferior lateral, basal, and medial quadrants. The resulting compartments are easily identified on postoperative magnetic resonance images obtained along the coronal plane. Resection within each compartment is noted as none (0), partial (1), or complete (2). An index of resection can then be computed for superior lateral, inferior lateral, basal, and medial quadrants and for the whole temporal lobe. Potentially, this technique can be used to evaluate surgical failures and correlate extent of resection with surgical outcome and neurologic complications. It also allows objective and semiquantitative comparison of surgical approaches practiced at different institutions.


Fs-laser ablation of teeth is temperature limited and provides information about the ablated components.
The goal of this work is to investigate the thermal effects of femtosecond laser (fs-laser) ablation for the removal of carious dental tissue. Additional studies identify different tooth tissues through femtosecond laser induced breakdown spectroscopy (fsLIBS) for the development of a feedback loop that could be utilized during ablation in a clinical setting. Scanning Election Microscope (SEM) images reveal that minimal morphological damages are incurred at repetition rates below the carbonization threshold of each tooth tissue. Thermal studies measure the temperature distribution and temperature decay during laser ablation and after laser cessation, and demonstrate that repetition rates at or below 10kHz with a laser fluence of 40 J/cm2 would inflict minimal thermal damage on the surrounding nerve tissues and provide acceptable clinical removal rates. Spectral analysis of the different tooth tissues is also conducted and differences between the visible wavelength fsLIBS spectra are evident, though more robust classification studies are needed for clinical translation. These results have initiated a set of precautionary recommendations that would enable the clinician to utilize femtosecond laser ablation for the removal of carious lesions while ensuring that the solidity and utility of the tooth remain intact.


Double-blind randomised trial of Org 10172 low-molecular-weight heparinoid in prevention of deep-vein thrombosis in thrombotic stroke.
In a double-blind, randomised trial Org 10172 low-molecular-weight (LMW) heparinoid was compared with placebo in the prevention of deep-vein thrombosis in patients with acute thrombotic stroke. Prophylaxis was started within 7 days of the onset of stroke with a loading dose of 1000 anti-factor-Xa units intravenously followed by a fixed dose of 750 anti-factor-Xa units twice a day subcutaneously; it was continued for 14 days or until hospital discharge, if earlier. 50 patients were randomised to receive Org 10172 and 25 to receive placebo. All patients underwent surveillance with I125-fibrinogen leg scanning and impedance plethysmography. Venography was carried out if either test became positive. Venous thrombosis occurred in 2 of 50 patients (4.0%) given Org 10172 and 7 of 25 patients (28.0%) given placebo (p = 0.005); the corresponding rates of proximal-vein thrombosis were 0% and 16%, respectively (p = 0.01). There was one major haemorrhage in the Org 10172 group and one minor bleed in the placebo group.


Biased movement drives local cryptic coloration on distinct urban pavements.
Explanations of how organisms might adapt to urban environments have mostly focused on divergent natural selection and adaptive plasticity. However, differential habitat choice has been suggested as an alternative. Here, we test for habitat choice in enhancing crypsis in ground-perching grasshoppers colonizing an urbanized environment, composed of a mosaic of four distinctly coloured substrates (asphalt roads and adjacent pavements). Additionally, we determine its relative importance compared to present-day natural selection and phenotypic plasticity. We found that grasshoppers are very mobile, but nevertheless approximately match the colour of their local substrate. By manipulating grasshopper colour, we confirm that grasshoppers increase the usage of those urban substrates that resemble their own colours. This selective movement actively improves crypsis. Colour divergence between grasshoppers on different substrates is not or hardly owing to present-day natural selection, because observed mortality rates are too low to counteract random substrate use. Additional experiments also show negligible contributions from plasticity in colour. Our results confirm that matching habitat choice can be an important driver of adaptation to urban environments. In general, studies should more fully incorporate that individuals are not only selective targets (i.e. selected on by the environment), but also selective agents (i.e. selecting their own environments).


Isotopic Analysis of Lead at Ultratrace Levels Using Thermal Ionization Mass Spectrometry (TIMS) Coupled with the Continuous Heating Method: Optimization of the Data Integration Range and Method.
This study systematically and experimentally evaluates data integration methods for the isotopic analysis of Pb at ultratrace levels using thermal ionization mass spectrometry (TIMS) with a continuous heating method. The evaluation utilized a certified reference material of Pb (SRM 981). The experimental evaluations encompass different data calculation methods (methods I, II, and III) and integration ranges (full, over 1%, 25%, and 75%). Method I, in which isotope ratios were calculated based on summed ion signal intensities compensating for mass fractionation, was consistent with the certified values for 10 and 1 ng standard samples across all integration ranges. For 100 pg samples, full range calculations failed for specific isotope ratios, but reduced ranges (over 1%, 25%, and 75%) yielded values overlapping with certified ones. Method II, in which isotope ratios were calculated by averaging the precalculated isotope ratios, exhibited inferior performance compared to method I. Method III, using weighted averaging to reduce anomalous values, showed results consistent with those of method I but was recommended only for single measurements. An integration range of over 1% or 25% is preferred to exclude anomalies while compensating for mass fractionation. The optimized method was validated by comparing two different instruments used for the isotopic analysis of the reference material. The enhanced accuracy and precision provide valuable insights for researchers working in ultratrace-level Pb isotopic analysis using TIMS.


Different characteristics of solubilized lactogen receptors from livers of pregnant and non-pregnant female rats.
Receptors specific for lactogenic hormones were solubilized by 1% (v/v) Triton X-100 from the crude particulate membrane fraction of livers of pregnant and non-pregnant female rats and the characteristics of both preparations were compared. Human (125)I-labelled somatotropin was used for binding studies of lactogenic hormone. The solubilized receptor retained most of the characteristics noted in the particulate fraction. The binding of human (125)I-labelled somatotropin to the solubilized receptor is a saturable process, depending on temperature and time. Scatchard analysis of displacement curves revealed similar affinity constants ranging from 1.02 x 10(9) to 1.20 x 10(9) 1/mol, while the binding capacity was 4.5 times greater in the pregnant rat livers than in the non-pregnant female rat livers. The receptors for human (125)I-labelled somatotropin from livers of non-pregnant and pregnant female rats were equally adsorbed onto a concanavalin-A-Sepharose column and were dissociated from the column with alpha-methyl-d-glucoside or alpha-methyl-d-mannoside in the same manner. By gel filtration on Sepharose 6B, however, the molecular sizes of the hepatic receptors were found to be different. The apparent M(r) value was approx. 270000 with a Stokes' radius of 5.5nm in the non-pregnant female rats and approx. 330000 with a Stokes' radius of 5.5nm in the pregnant rats. Furthermore, isoelectric-focusing experiments showed that a major part of the receptor from the non-pregnant female rat livers had a neutral pI (7.0-8.5), whereas that from pregnant-rat livers had an acidic pI (4.2-4.7). These data suggest that the increase in the lactogenic binding capacity in rat liver membranes during pregnancy may be associated with marked changes of the physicochemical properties of the receptors.


Engineering a CEACAM1 Variant with the Increased Binding Affinity to TIM-3 Receptor
T-cell immunoglobulin domain and mucin domain-3 (TIM-3) is an inhibitory receptor expressed in a variety of cells, including dendritic cells, T-helper 1 lymphocytes, and natural killer cells. Binding of this protein to its ligand, carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), causes T-cell exhaustion, a specific condition in which effector T cells lose their ability to proliferate and produce cytokines. Blocking this inhibitory receptor is known to be an effective strategy for treating cancer and other related diseases. Therefore, in this study, in order to block the inhibitory receptor of TIM-3, we designed and produced recombinantly a protein with a high binding affinity to this receptor. The extracellular domain of CEACAM1 involved in binding to TIM-3 was mutated using R script to obtain a variant with the increased binding affinity to TIM-3. The binding energy of the mutant protein was calculated using the FoldX module. Finally, after recombinant production of the most appropriate CEACAM1 variant (variant 39) in E. coli, its secondary structure was determined by CD spectroscopy. The binding free energy between variant 39 and TIM-3 decreased from -5.63 to -14.49 kcal/mol, indicating an increased binding affinity to the receptor. Analysis of the secondary structure of this variant also showed that the mutation did not significantly alter the structure of the protein. Our findings suggest that variant 39 could bind to TIM-3 with a higher binding affinity than the wild-type, making it a proper therapeutic candidate for blocking TIM-3.


The evaluation and treatment of adults with gastroesophageal reflux disease.
Gastroesophageal reflux disease (GERD) is defined as symptoms or tissue damage that results from the abnormal reflux of gastric contents into the esophagus. A systematic review of population-based studies estimates that heartburn or regurgitation symptoms occur in 21% to 59% of the population during a given year. The frequency of GERD in specific populations is provided in Table 1. Although only 1 in 5 patients with upper intestinal symptoms that occur at least weekly seeks medical attention, nearly 1% of all visits to a family physician's office are for GERD or related conditions. GERD significantly affects the quality of patients' lives. In a survey of patients presenting for upper endoscopy with symptoms of at least 3 months' duration, those with a diagnosis of GERD reported low scores at baseline for general well-being. Fortunately, follow-up data reported 4 weeks after treatment note improvement in gastrointestinal symptoms, general well-being, general health, vitality, and depression.


Is infertility resolution associated with a change in women's well-being?
Is giving birth associated with improved subjective well-being among involuntarily childless women? Resolution of infertility is associated with increased life satisfaction and self-esteem, but not with a decrease in depressive symptoms. Cross-sectional data and studies of treatment-seekers show that infertility is associated with lower subjective well-being. Childless women with infertility tend to report lower subjective well-being than women who experience secondary infertility, but a prospective study using a random sample of involuntarily childless women over time has not previously been conducted. The sample for the current study includes all women without children who met medical criteria for infertility or perceived a fertility problem (N = 283) at baseline and who were interviewed in both waves (3 years apart) of the National Survey of Fertility Barriers (NSFB), in a random-digit dialing telephone survey. It is therefore possible to explore here whether there are differences in the association of infertility resolution and subjective well-being among women who do and do not perceive themselves as having a fertility problem. Depressive symptoms (as measured by the Center for Epidemiologic Studies-Depression Scale), self-esteem (as measured by a modified version of the Rosenberg Self-esteem Scale) and life satisfaction (as measured by a modified version of the Satisfaction with Life Scale) were assessed for all 283 participants at both waves. For all three variables, change scores of 47 involuntarily childless women who resolved their infertility through a live birth were compared to the scores for the 236 women who remained childless. A number of variables shown to be associated with subjective well-being among infertile women were included as controls. No relationship between infertility resolution and change in depressive symptoms was observed (b = -0.04; P > 0.05). Involuntarily childless women who resolved their infertility improved in self-esteem (b = 0.74; P < 0.01) and life satisfaction (b = 1.06; P < 0.01). Women were measured at only two time points. Only 47 women had a live birth between waves. While it is common practice to make causal interpretations based on panel data, such interpretations should be made with caution. In addition, the NSFB was conducted in the USA where medical expenditures are high and most fertility treatment expenses are not covered by insurance. Thus it may not be possible to generalize the findings to other modern industrialized societies. Knowing that resolution of infertility is associated with improved subjective well-being is important for infertile couples and infertility professionals alike. This research was supported in part by NICHD grant R01-HD044144 and NIGMS grant P20-GM109097 from the National Institutes of Health. The authors have no competing interests.


Smoking and the Risk of Hospitalization for Symptomatic Diverticular Disease: A Population-Based Cohort Study from Sweden.
Current studies reporting on the risk of smoking and development of symptomatic diverticular disease have reported conflicting results. The aim of this study was to investigate the association between smoking and symptomatic diverticular disease. This is a cohort study : Information was derived from the Swedish Construction Workers Cohort 1971-1993. Patients were selected from construction workers in Sweden. The primary outcome measured was the development of symptomatic diverticular disease and complicated diverticular disease (abscess and perforation) as identified in the Swedish Hospital Discharge Register. Adjusted relative risks of symptomatic diverticular disease according to smoking status were estimated by using negative binomial regression analysis. In total, the study included 232,685 men and 14,592 women. During follow-up, 3891 men and 318 women had a diagnosis of later symptomatic diverticular disease. In men, heavy smokers (≥15 cigarettes a day) had a 1.6-fold increased risk of developing symptomatic diverticular disease compared with nonsmokers (adjusted relative risk, 1.56; 95% CI, 1.42-1.72). There was evidence of a dose-response relationship, because moderate and ex-smokers had a 1.4- and 1.2-fold increased risk compared with nonsmokers (adjusted relative risk, 1.39; 95% CI, 1.27-1.52 and adjusted relative risk, 1.14; 95% CI, 1.04-1.27). These relationships were similar in women, but the risk estimates were less precise owing to smaller numbers. Male ever-smokers had a 2.7-fold increased risk of developing complicated diverticular disease (perforation/abscess) compared with nonsmokers (adjusted relative risks, 2.73; 95% CI, 1.69-4.41). We were unable to account for other confounding variables such as comorbidity, prescription medication, or lifestyle factors. Smoking is associated with symptomatic diverticular disease in both men and women and with an increased risk of developing complicated diverticular disease.


Synchrotron Time-Lapse Imaging of Lignocellulosic Biomass Hydrolysis: Tracking Enzyme Localization by Protein Autofluorescence and Biochemical Modification of Cell Walls by Microfluidic Infrared Microspectroscopy.
Tracking enzyme localization and following the local biochemical modification of the substrate should help explain the recalcitrance of lignocellulosic plant cell walls to enzymatic degradation. Time-lapse studies using conventional imaging require enzyme labeling and following the biochemical modifications of biopolymers found in plant cell walls, which cannot be easily achieved. In the present work, synchrotron facilities have been used to image the enzymatic degradation of lignocellulosic biomass without labeling the enzyme or the cell walls. Multichannel autofluorescence imaging of the protein and phenolic compounds after excitation at 275 nm highlighted the presence or absence of enzymes on cell walls and made it possible to track them during the reaction. Image analysis was used to quantify the fluorescence intensity variations. Consistent variations in the enzyme concentration were found locally for cell cavities and their surrounding cell walls. Microfluidic FT-IR microspectroscopy allowed for time-lapse tracking of local changes in the polysaccharides in cell walls during degradation. Hemicellulose degradation was found to occur prior to cellulose degradation using a Celluclast® preparation. Combining the fluorescence and FT-IR information yielded the conclusion that enzymes did not bind to lignified cell walls, which were consequently not degraded. Fluorescence multiscale imaging and FT-IR microspectroscopy showed an unexpected variability both in the initial biochemical composition and the degradation pattern, highlighting micro-domains in the cell wall of a given cell. Fluorescence intensity quantification showed that the enzymes were not evenly distributed, and their amount increased progressively on degradable cell walls. During degradation, adjacent cells were separated and the cell wall fragmented until complete degradation.


Atherogenic indices and prehypertension in obese and non-obese children.
We evaluate the relationship between different lipoproteins and atherogenic indices with pre-hypertension in 297 obese and 942 non-obese children with Tanner stage 1 enrolled in a multicentre, community-based cross-sectional study. Height, weight, fasting glucose and insulin levels, total cholesterol, triglycerides, high-density lipoprotein cholesterol (HDL-c), low-density lipoprotein cholesterol (LDL-c), non-HDL-c, LDL/HDL-c, triglycerides/cholesterol and total cholesterol/HDL-c ratios were measured. Mean age was 8.4 ± 1.2 years; pre-hypertension was identified in 104 (8.4%) participants, 46 (15.5%) obese and 58 (6.1%) non-obese children. The pre-hypertensive non-obese children show a high proportion of family history of hypertension (41.6 and 24.7%, p = 0.002) and elevation of insulin at a relatively low body mass index. The triglycerides:HDL-c ratio, but not other lipoproteins or atherogenic indices, was associated with pre-hypertension in obese (1.15, 95% confidence intervals 1.06-1.26) and non-obese children (1.38 95% confidence intervals 1.22-1.57). The triglycerides:HDL-c ratio is related to pre-hypertension in children; the family history of hypertension seems to be a risk factor in developing pre-hypertension.


Designing the operative suite.
The planning of an operative suite involves many considerations, often of a highly technical nature. Requirements have become so diversified and have been elaborated so rapidly that standardization of requirements cannot be anticipated. The concept of grouping interdependent departments has brought the suite down to lower floors. Rooms have become larger to accommodate more monitoring and other equipment, and many more ancillary rooms have been found necessary. A wide wing with double or peripheral corridors is preferable. Air sterilization can be achieved by several methods. The doctors' dressing room is often a danger point in bacterial control and needs redesigning. Patient monitoring is increasing and some features can be built in. TV observation and teaching have tremendous potential but have not been adopted as widely as was anticipated some years ago. If a department needs extensive enlargement, it is much more satisfactory and usually cheaper to construct a new suite in another location.


The protective function of neutrophil elastase inhibitor in liver ischemia/reperfusion injury.
BACKGROUND.: A neutrophil elastase (NE) inhibitor, Sivelestat, has been approved for the treatment of acute lung injury associated with systemic inflammation in humans. Some reports have also shown its protective effects in liver inflammatory states. We have recently documented the importance of NE in the pathophysiology of liver ischemia/reperfusion injury, a local Ag-independent inflammation response. This study was designed to explore putative cytoprotective functions of clinically available Sivelestat in liver ischemia/reperfusion injury. METHODS.: Partial warm ischemia was produced in the left and middle hepatic lobes of C57BL/6 mice for 90 min, followed by 6 or 24 hr of reperfusion. The mice were given Sivelestat (100 mg/kg, subcutaneous) at 10 min before ischemia, 10 min before reperfusion, and at 1 and 3 hr of reperfusion thereafter. RESULTS.: Sivelestat treatment significantly reduced serum alanine aminotransferase levels and NE activity, when compared with controls. Histological liver examination has revealed that unlike in controls, Sivelestat ameliorated the hepatocellular damage and decreased local neutrophil activity and infiltration. The expression of proinflammatory cytokines (tumor necrosis factor-alpha and interleukin-6), chemokines (CXCL-1, CXCL-2, and CXCL-10), and toll-like receptor 4 was significantly reduced in the treatment group, along with diminished apoptosis through caspase-3 pathway. Moreover, in vitro studies confirmed downregulation of proinflammatory cytokine and chemokine programs in mouse macrophage cell cultures, along with depression of innate toll-like receptor 4 signaling. CONCLUSION.: Sivelestat-mediated NE inhibition may represent an effective therapeutic option in liver transplantation and other inflammation disease states.


Implementation of CsLIS/NES in linalool biosynthesis involves transcript splicing regulation in Camellia sinensis.
Volatile terpenoids produced in tea plants (Camellia sinensis) are airborne signals interacting against other ecosystem members, but also pleasant odorants of tea products. Transcription regulation (including transcript processing) is pivotal for plant volatile terpenoid production. In this study, a terpene synthase gene CsLIS/NES was recovered from tea plants (C. sinensis cv. "Long-Men Xiang"). CsLIS/NES transcription regulation resulted in 2 splicing forms: CsLIS/NES-1 and CsLIS/NES-2 lacking a 305 bp-fragment at N-terminus, both producing (E)-nerolidol and linalool in vitro. Transgenic tobacco studies and a gene-specific antisense oligo-deoxynucleotide suppression applied in tea leaves indicated that CsLIS/NES-1, localized in chloroplasts, acted as linalool synthase, whereas CsLIS/NES-2 localized in cytosol, functioned as a potential nerolidol synthase, but not linalool synthase. Expression patterns of the 2 transcript isoforms in tea were distinctly different and responded differentially to the application of stress signal molecule methyl jasmonate. Leaf expression of CsLIS/NES-1, but not CsLIS/NES-2, was significantly induced by methyl jasmonate. Our data indicated that distinct transcript splicing regulation patterns, together with subcellular compartmentation of CsLIS/NE-1 and CsLIS/NE-2 implemented the linalool biosynthesis regulation in tea plants in responding to endogenous and exogenous regulatory factors.


A Comparison of Hemodynamic and Clinical Outcomes After Transcatheter Versus Surgical Therapy in Adults in Coarctation of Aorta.
Transcatheter stent therapy provides similar acute reduction in coarctation of aorta (COA) gradient and systolic blood pressure (SBP) as compared with surgery. However, there are limited data comparing mid-term outcomes after transcatheter vs surgical therapy for COA. The purpose of this study was to compare temporal changes in Doppler COA gradient and SBP after transcatheter stent therapy versus surgical therapy for COA. A retrospective study of COA patients (≥18 years old) undergoing transcatheter stent therapy or surgical therapy at Mayo Clinic in Rochester, Minnesota from 2000-2018 was performed. Linear regression analyses were used to compare temporal changes in Doppler gradient and SBP between the 2 groups. Propensity matching was used to adjust for between-group differences in clinical and anatomic characteristics. A total of 44 and 128 patients underwent transcatheter and surgical therapy, respectively; there were no significant between-group differences in the anatomy of the thoracic aorta. Both groups had similar acute reduction in Doppler peak gradient (P=.66), mean gradient (P=.41), SBP (P=.22), and upper-to-lower extremity SBP (ULE-SBP) gradient (P=.69). The median follow-up was 46 months (interquartile range, 27-81 months) and 63 months (interquartile range, 41-94 months) in the transcatheter and surgical groups, respectively. There were no significant between-group differences in the temporal change in Doppler peak gradient (P-interaction=.06), mean gradient (P-interaction=.15), SBP (P-interaction=.20), and ULE-SBP gradient (P-interaction=.51). These favorable short- and mid-term outcome data support the use of transcatheter therapy as an alternative to surgery in adults with COA. Further studies are required to determine if these favorable outcomes are maintained on long-term follow-up.


Effect of astaxanthin supplementation on inflammation and cardiac function in BALB/c mice.
Astaxanthin is an antioxidant with immunomodulatory, anti-inflammatory and anticancer properties. This study evaluated the use of dietary astaxanthin to decrease oxidative stress and improve cardiac function, thereby providing a potential cardioprotective supplement. Female BALB/c mice (8 weeks of age) were fed a semi-synthetic diet containing 0, 0.02 or 0.08% astaxanthin for 8 weeks. Cardiac function was assessed by echocardiography bi-weekly, and blood and tissue samples were collected at 8 weeks. Plasma astaxanthin concentrations increased (p<0.05) dose-dependently to 0.5 and 4 mumol/l in the astaxanthin-supplemented mice. Blood glutathione concentrations and lymphocyte mitochondrial membrane potential were not significantly affected by astaxanthin treatment. However, mice fed 0.08% astaxanthin had higher (p<0.05) heart mitochondrial membrane potential and contractility index compared to the control group. These results support the possible use of dietary astaxanthin for cardiac protection.


[The beginning of human life: ethical and legal perspectives in the context of biotechnological progress].
Questions concerning the beginning of human life have pervaded society since antiquity. In the post-modern world, scientific and technological advances have fueled discussions on the issue, such that debates previously concentrated on abortion now also focus on biotechnological interventions. The article addresses the latter, reflecting on the extent to which human dignity can be considered a (hermeneutic) reference in establishing ethical and legal parameters for biotechnological advances in the definition of the beginning of human life. The study's method was critical hermeneutic ethics, with ethics at the center of the process of understanding and interpretation, observing the contours of facticity. No consensus was found on the beginning of human life, so it is essential to engage in dialogue with the new reality resulting from biotechnological advances in the process of defining ethical and legal principles for protecting the embryo and human nature, with human dignity as the reference.


Spectral multivariate calibration with wavelength selection using variants of Tikhonov regularization.
Tikhonov regularization (TR) is a general method that can be used to form a multivariate calibration model and numerous variants of it exist, including ridge regression (RR). This paper reports on the unique flexibility of TR to form a model using full wavelengths (RR), individually selected wavelengths, or multiple bands of selected wavelengths. Of these three TR variants, the one based on selection of wavelength bands is found to produce lower prediction errors. As with most wavelength selection algorithms, the model vector magnitude indicates that this error reduction comes with a potential increase in prediction uncertainty. Results are presented for near-infrared, ultraviolet-visible, and synthetic spectral data sets. While the focus of this paper is wavelength selection, the TR methods are generic and applicable to other variable-selection situations.


GAS5 promotes podocyte injury in sepsis by inhibiting PTEN expression.
To investigate the potential role of long noncoding RNA (lncRNA) growth arrest specific transcript 5 (GAS5) in sepsis-induced podocyte injury and its underlying mechanism. The sepsis model was established by lipopolysaccharide (LPS) induction in podocytes. The expression levels of Nephrin and GAS5 were detected by quantitative Real-time polymerase chain reaction (qRT-PCR) after LPS induction in podocytes for 12 h, 24 h and 36 h, respectively. Western blot was used to detect the expression level of Nephrin in sepsis-induced podocytes. The mRNA expressions of GAS5 and Nephrin in podocytes were detected after transfection of GAS5 siRNA. Albumin influx in podocytes after GAS5 knockdown was detected by Transwell assay. Western blot was used to detect the protein expression of Snail in sepsis after GAS5 knockdown. The target gene of GAS5 was predicted by bioinformatics analysis. QRT-PCR and Western blot were used to detect the protein and mRNA levels of PTEN (phosphatase and tensin homolog deleted on chromosome ten). Nephrin expression and the albumin inflow after PTEN knockdown were then measured. The expression of PI3K/AKT/GSK3β was also detected after GAS5 was downregulated while PTEN was upregulated. LPS stimulation downregulated the mRNA expression of Nephrin in podocytes and achieved the lowest level at 24 h. The protein expression change of Nephrin was consistent with its mRNA expression. In the septic state, the albumin influx of podocytes remarkably increased, but the function of podocyte barrier was weakened. Besides, GAS5 expression decreased in a time-dependent manner in LPS-induced podocytes. After GAS5 knockdown by siRNA, Nephrin expression and the function of podocyte barrier were significantly reduced. Snail expression was also upregulated in septic state, and GAS5 knockdown increased the expressions of phosphorylated Snail and PI3K/AKT/GSK3β. After knockdown of GAS5, the mRNA and protein levels of PTEN significantly decreased, which was contract to the expression of Snail. However, overexpression of PTEN could reverse the promotive effect of GAS5 on PI3K/AKT activation. GAS5 expression decreased in sepsis-induced podocyte injury, and GAS5 was involved in regulating sepsis-induced podocyte injury by reducing PTEN expression.


The (non-)replicability of regulatory resource depletion: A field report employing non-invasive brain stimulation.
Cognitive effort and self-control are exhausting. Although evidence is ambiguous, behavioural studies have repeatedly suggested that control-demanding tasks seem to deplete a limited cache of self-regulatory resources leading to performance degradations and fatigue. While resource depletion has indirectly been associated with a decline in right prefrontal cortex capacity, its precise neural underpinnings have not yet been revealed. This study consisted of two independent experiments, which set out to investigate the causal role of the right dorsolateral prefrontal cortex (DLPFC) in a classic dual phase depletion paradigm employing non-invasive brain stimulation. In Experiment 1 we demonstrated a general depletion effect, which was significantly eliminated by anodal transcranial Direct Current Stimulation to the right DLPFC. In Experiment 2, however, we failed to replicate the basic psychological depletion effect within a second independent sample. The dissimilar results are discussed in the context of the current 'replication crisis' and suggestions for future studies are offered. While our current results do not allow us to firmly argue for or against the existence of resource depletion, we outline why it is crucial to further clarify which specific external and internal circumstances lead to limited replicability of the described effect. We showcase and discuss the current inter-lab replication problem based on two independent samples tested within one research group (intra-lab).


Effect of addition of inhaled salmeterol to the treatment of moderate-to-severe asthmatics uncontrolled on high-dose inhaled steroids. European Respiratory Study Group.
The aim of this study was to assess the efficacy and safety of inhaled salmeterol 100 micrograms b.d. (SM) versus inhaled salbutamol 400 micrograms q.d.s. (SB), both via the Diskhaler, when added to concurrent treatment, in asthmatic patients who were not controlled on high doses of inhaled steroids (> or = 1,500 micrograms beclomethasone dipropionate (BDP) or equivalent daily). This was a multicentre, parallel group, double-blind study in which 190 patients with a forced expiratory volume in one second (FEV1) or peak expiratory flow rate (PEFR) of 30-75% predicted and 15% reversibility to inhaled bronchodilator were randomized to treatment for 6 weeks. In the SM group, morning PEFR increased from 281 to 315 L-min-1 during treatment and in the SB group from 311 to 315 L.min-1 (p < 0.001). The SM group showed significantly better reduction in diurnal variation, from 39 to 22 L.min-1 during treatment, than the SB group (34 to 37 L.min-1) (p < 0.001). There was a significantly greater improvement in FEV1 in the SM group (from 1.63 to 1.85 L) than in the SB group (from 1.79 to 1.84 L). The SM group had significantly more symptom-free nights than the SB group (p < 0.001), and also more "rescue-free" nights (p = 0.04). The adverse event profile was similar in both groups. This study indicates that in asthmatic patients, not controlled on high-dose inhaled steroids, inhaled salmeterol 100 micrograms b.d. significantly improves lung function and reduces asthma symptoms.


Characterization of mouse fibronectin alternative mRNAs reveals an unusual isoform present transiently during liver development.
Fibronectins are found in many extracellular matrices as well as being abundant plasma proteins. The plasma isoforms of fibronectin, which are synthesized in the adult by liver hepatocytes, differ from those derived from most other cells and tissues due to alternative mRNA splicing. Studies in several vertebrates have indicated that FN alternative splicing is regulated spatially and temporally during development. The mouse represents an attractive organism in which to study the regulation of fibronectin splicing during development, but the patterns of fibronectin alternative splicing were not known for this species. Mouse fibronectin cDNA clones were isolated and sequenced, revealing > 95% identity with rat fibronectin at the amino acid level; all three segments that undergo alternative splicing are well conserved. RNase protection and RT-PCR were used to determine the patterns of alternative splicing that occur in fibroblasts and adult liver, sources of cellular and plasma fibronectins. Only A-B-mRNAs were detected in liver, and three V region variants were observed, corresponding to the protein isoforms V120, V95, and V0. Fibroblasts produced mRNAs that were heterogeneous for A and B splicing, but all RNAs contained V120. These patterns contrast with the embryonic form (B+A+V120). Characterization of fibronectin mRNAs from livers of fetal and newborn mice revealed that a significant level of B+ mRNA was present throughout late gestation, declining at birth. Little A+ mRNA was present, and the adult liver V region pattern was observed at all stages. Thus, fibronectin splicing changes during liver development are noncoordinate. One consequence of this temporal regulation is the transient synthesis of B+ mRNAs, including a novel isoform, B+A-V0.


Effects of foot-and-mouth disease virus nonstructural proteins on the structure and function of the early secretory pathway: 2BC but not 3A blocks endoplasmic reticulum-to-Golgi transport.
Infection of cells by picornaviruses leads to the generation of intracellular membrane vesicles. The expression of poliovirus (PV) 3A protein causes swelling of the endoplasmic reticulum (ER) and inhibition of protein trafficking between the ER and the Golgi apparatus. Here, we report that the nonstructural proteins of a second picornavirus, foot-and-mouth disease virus (FMDV), also perturb the secretory pathway. FMDV proteins 3A, 2B, 2C, and 2BC expressed alone in cells were recovered from crude membrane fractions, indicating membrane association. Immunofluorescence microscopy showed that 3A was located in a reticular structure and 2B was located in the ER, while 2C was located in both the ER and the bright punctate structures within the Golgi apparatus. 2BC gave punctate cytoplasmic staining and also caused accumulation of ER proteins in large vesicular structures located around the nuclei. The effect of the FMDV proteins on the trafficking of the vesicular stomatitis virus glycoprotein (G protein) from the ER to the cell surface was determined. Unlike its PV counterpart, the 3A protein of FMDV did not prevent trafficking of the G protein to the cell surface. Instead, surface expression of the G protein was blocked by 2BC, with retention of the G protein in a modified ER compartment staining for 2BC. The results suggest that the nonstructural proteins of different picornaviruses may vary in their ability to perturb the secretory pathway. Since FMDV 2BC can block the delivery of proteins to the cell surface, it may, as shown for PV 3A, play a role in immune evasion and contribute to the persistent infections observed in ruminants.


Value of DNA integrity assays for fertility evaluation.
DNA damage in the male germ line is associated with failed fertilisation, impaired preimplantation development and poor pregnancy outcomes, whether the insemination is natural or artificial. It is also apparent that DNA damage in spermatozoa is associated with adverse impacts on the health and wellbeing of children. This may be particularly important in the case of conceptions involving intracytoplasmic sperm injection. With this technique DNA damaged spermatozoa that would, under physiological circumstances, be excluded from the conception process are able to initiate pregnancies. Although the oocyte actively repairs the DNA damage brought into the zygote by the fertilising spermatozoon, errors in this repair process would generate mutations that might in turn be linked to the increased incidence of dominant genetic disease and childhood cancer seen in the offspring of fathers possessing DNA damaged spermatozoa. Significantly, the incidence of such mutations is so low that it might be several generations before the full consequences of using DNA damaged spermatozoa in assisted conception cycles are realised. The factors modulating DNA damage in the male germ line are complex and largely unresolved. They involve advanced paternal age, exposure to xenobiotics and male genital tract infection. The types of damage being measured by the assays used in most laboratories (SCSA, Comet and TUNEL) are also uncertain. Molecular characterization of this DNA damage might provide insights into the underlying aetiologies, facilitate the development of optimised diagnostic methods for its detection and suggest logical avenues to pursue for its correction and ultimate prevention.


Sulfated steroids as natural ligands of mouse pheromone-sensing neurons.
Among mice, pheromones and other social odor cues convey information about sex, social status, and identity; however, the molecular nature of these cues is essentially unknown. To identify these cues, we screened chromatographic fractions of female mouse urine for their ability to cause reproducible firing rate increases in the pheromone-detecting vomeronasal sensory neurons (VSNs) using multielectrode array (MEA) recording. Active compounds were found to be remarkably homogenous in their basic properties, with most being of low molecular weight, moderate hydrophobicity, low volatility, and possessing a negative electric charge. Purification and structural analysis of active compounds revealed multiple sulfated steroids, of which two were identified as sulfated glucocorticoids, including corticosterone 21-sulfate. Sulfatase-treated urine extracts lost >80% of their activity, indicating that sulfated compounds are the predominant VSN ligands in female mouse urine. As measured by MEA recording, a collection of 31 synthetic sulfated steroids triggered responses 30-fold more frequently than did a similarly sized stimulus set containing the majority of all previously reported VSN ligands. Collectively, VSNs detected all major classes of sulfated steroids, but individual neurons were sensitive to small variations in chemical structure. VSNs from both males and females detected sulfated steroids, but knock-outs for the sensory transduction channel TRPC2 did not detect these compounds. Urine concentrations of the two sulfated glucocorticoids increased many fold in stressed animals, indicating that information about physiological status is encoded by the urine concentration of particular sulfated steroids. These results provide an unprecedented characterization of the signals available for chemical communication among mice.


Type-II/type-II band alignment to boost spatial charge separation: a case study of g-C3N4 quantum dots/a-TiO2/r-TiO2 for highly efficient photocatalytic hydrogen and oxygen evolution.
Efficient spatial charge separation and transfer that are critical factors for solar energy conversion primarily depend on the energetic alignment of the band edges at interfaces in heterojunctions. Herein, we first report that constructing a 0D/0D type-II(T-II)/T-II heterojunction is an effective strategy to ingeniously achieve long-range charge separation by taking a ternary heterojunction of TiO2 and graphitic carbon nitride (g-C3N4) as a proof-of-concept. Incorporating g-C3N4 quantum dots (QCN), as the third component, into the commercial P25 composed of anatase (a-TiO2) and rutile (r-TiO2) can be realized via simply mixing the commercially available Degussa P25 and QCN solution followed by heat treatment. The strong coupling and matching band structures among a-TiO2, r-TiO2 and QCN result in the construction of novel T-II/T-II heterojunctions, which would promote the spatial separation and transfer of photogenerated electrons and holes. Moreover, QCN plays a key role in reinforcing light absorption. Particularly, the unique 0D/0D architecture possesses the advantages of abundant active sites for the photocatalytic reaction. As a result, the optimized QCN/a-TiO2/r-TiO2 heterojunctions exhibit enhanced photocatalytic H2 and O2 evolution, especially the hydrogen evolution rate (49.3 μmol h-1) is 11.7 times that of bare P25 under visible light irradiation, and sufficient catalytic stability as evidenced by the recycling experiments. The remarkably enhanced photocatalytic activity can be attributed to the synergistic effects of the energy level alignment at interfaces, the dimensionality and component of the heterojunctions. This work provides a stepping stone towards the design of novel heterojunctions for photocatalytic water splitting.


The operating surgeon is an independent predictor of chest tube drainage following cardiac surgery.
Bleeding into the chest is a major cause of blood transfusion and adverse outcomes following cardiac surgery. The authors investigated predictors of bleeding following cardiac surgery to identify potentially correctable factors. Data were retrieved from the medical records of patients undergoing cardiac surgery over the period of 2002 to 2008. Multivariate analysis was used to identify the independent predictors of chest tube drainage. Tertiary hospital. Two thousand five hundred seventy-five patients. Cardiac surgery. The individual operating surgeon was independently associated with the extent of chest tube drainage. Other independent factors included internal mammary artery grafting, cardiopulmonary bypass time, urgency of surgery, tricuspid valve surgery, redo surgery, left ventricular impairment, male gender, lower body mass index and higher preoperative hemoglobin levels. Both a history of diabetes and administration of aprotinin were associated with reduced levels of chest tube drainage. The individual operating surgeon was an independent predictor of the extent of chest tube drainage. Attention to surgeon-specific factors offers the possibility of reduced bleeding, fewer transfusions, and improved patient outcomes.


Some properties of pig kidney-cortex aldehyde reductase.
Aldehyde reductase was purified from pig kidney cortex to homogeneity by a new procedure. The molecular weight of the enzyme was estimated by sedimentation equilibrium to be 43 700 and by gel electrophoresis in the presence of sodium dodecyl sulphate to be 41 700. The enzyme is clearly a monomer. The enzyme preparation contained no significant quantities of zinc, manganese or copper and had no essential histidine or thiol groups. Changes in the absorption and fluorescence spectra of NADPH were observed on formation of the enzyme-NADPH complexes. Large changes in the fluorescence spectra were also observed in the presence of sodium barbitone or Warfarin. These changes were used as the basis of active-site titrations, which showed that the enzyme had one active site per molecule. The dissociation constants of NADPH and NADP+ from binary complexes with the enzyme were estimated in spectrophotometric titrations.


[Disruption of male meiosis in transgenic line tobacco res91].
The course of meiosis in male-sterile trasgenic tobacco line res91 has been analysed. Cytological analysis included visualization of the spindle and phragmoplast. Abnormal meiosis was characterized with 1) cytomixis; 2) deformation of nuclei at prophase 2 in part of the cells, and lack of spindle formation in such cells at M2; 3) desorientation of spindles in meiosis 2. This set of abnormalities allows to suppose certain disturbancies of cytoskeleton during male meiosis in res91.


Antiviral activity of RhoA-derived peptides against respiratory syncytial virus is dependent on formation of peptide dimers.
A synthetic peptide containing amino acids 77 to 95 of the intracellular GTPase RhoA has previously been shown to inhibit replication of respiratory syncytial virus (RSV) in cultured cells. We show that residues 80 to 90 of RhoA are sufficient for this activity and that the cysteine residue at position 83 is critical. Further studies with an optimal peptide sequence containing amino acids 80 to 94 of RhoA revealed that the antiviral potency of the peptide is dependent on the oxidation of cysteine 83. Size-exclusion chromatography and sedimentation equilibrium studies of the peptide comprising residues 80 to 94 revealed that it is capable of forming aggregates in both reduced and oxidized states. A peptide (83A) in which the cysteine residue is replaced by an alanine does not form dimers or higher-order aggregates and did not inhibit RSV replication at any concentration tested. These data indicate that formation of peptide multimers is necessary for the antiviral activities of RhoA-derived peptides and suggest that the observed antiviral activities of these peptides may be unrelated to the biological functions of their parent molecule.


Business forecasting methods: Impressive advances, lagging implementation.
Reliable forecasts are key to decisions in areas ranging from supply chain management to capacity planning in service industries. It is encouraging then that recent decades have seen dramatic advances in forecasting methods which have the potential to significantly increase forecast accuracy and improve operational and financial performance. However, despite their benefits, we have evidence that many organizations have failed to take up systematic forecasting methods. In this paper, we provide an overview of recent advances in forecasting and then use a combination of survey data and in-depth semi-structured interviews with forecasters to investigate reasons for the low rate of adoption. Finally, we identify pathways that could lead to the greater and more widespread use of systematic forecasting methods.


Hormesis, hotspots and emissions trading.
Instrument choice--the comparison of technology standards, performance standards, taxes and tradable permits--has been a major topic in environmental law and environmental economics. Most analyses assume that emissions and health effects are positively and linearly related. If they are not, this complicates the instrument choice analysis. This article analyses the effects of a nonlinear dose-response function on instrument choice. In particular, it examines the effects of hormesis (high-dose harm but low-dose benefit) on the choice between fixed performance standards and tradable emissions permits. First, the article distinguishes the effects of hormesis from the effects of local emissions. Hormesis is an attribute of the dose-response or exposure-response relationship. Hotspots are an attribute of the emissions-exposure relationship. Some pollutants may be hormetic and cause local emissions-exposure effects; others may be hormetic without causing local emissions-exposure effects. It is only the local exposure effects of emissions that pose a problem for emissions trading. Secondly, the article shows that the conditions under which emissions trading would perform less well or even perversely under hormesis, depend on how stringent a level of protection is set. Only when the regulatory standard is set at the nadir of the hormetic curve would emissions trading be seriously perverse (assuming other restrictive conditions as well), and such a standard is unlikely. Moreover, the benefits of the overall programme may justify the risk of small perverse effects around this nadir. Thirdly, the article argues that hotspots can be of concern for two distinct reasons, harmfulness and fairness. Lastly, the paper argues that the solution to these problems may not be to abandon market-based incentive instruments and their cost-effectiveness gains, but to improve them further by moving from emissions trading and emissions taxes to risk trading and risk taxes. In short, the article argues that hormesis does not pose a general obstacle to emissions trading or emissions taxes, but that in those cases where hormesis does pose such a problem, a shift toward risk trading or risk taxes would be the superior route.


Religious beliefs and climate change adaptation: A study of three rural South African communities.
This article argues that religious beliefs significantly influence a community's understanding and experience of climate change adaptation, indicating the need for an inclusion of such information in climate change adaptation education. Data were collected using the Q-method, whereby recurring statements were identified from semi-structured interviews with participants from three rural communities in the North-West province of South Africa: Ikageng, Ventersdorp and Jouberton. The research found that community members who regard themselves as religious (overall of the Christian faith) fall under two groups: the religious determinists or fatalists, who see climate as a natural process that is governed by God, and religious participants who deny this 'naturalness' and acknowledge humans' impact on the climate.


A Chlamydomonas outer arm dynein mutant missing the alpha heavy chain.
A novel Chlamydomonas flagellar mutant (oda-11) missing the alpha heavy chain of outer arm dynein but retaining the beta and gamma heavy chains was isolated. Restriction fragment length polymorphism analysis with an alpha heavy chain locus genomic probe indicated that the oda-11 mutation was genetically linked with the structural gene of the alpha heavy chain. In cross-section electron micrographs, the oda-11 axoneme lacked the outermost appendage of the outer arm, indicating that the alpha heavy chain should be located in this region in the wild-type outer arm. This mutant swam at 119 microns/s at 25 degrees C, i.e., at an intermediate speed between those of wild type (194 microns/s) and of oda-1 (62 microns/s), a mutant missing the entire outer dynein arm. The flagellar beat frequency (approximately 50 Hz) was also between those of wild type (approximately 60 Hz) and oda-1 (approximately 26 Hz). These results indicate that the outer dynein arm of Chlamydomonas can be assembled without the alpha heavy chain, and that the outer arm missing the alpha heavy chain retains partial function.


Model-based imputation of latent cigarette counts using data from a calibration study.
In addition to dichotomous measures of abstinence, smoking studies may use daily cigarette consumption as an outcome variable. These counts hold the promise of more efficient and detailed analyses than dichotomous measures, but present serious quality issues - measurement error and heaping - if obtained by retrospective recall. A doubly-coded dataset with a retrospective recall measurement (timeline followback, TLFB) and a more precise instantaneous measurement (ecological momentary assessment, EMA) serves as a calibration dataset, allowing us to predict EMA given TLFB and baseline factors. We apply this model to multiply impute precise cigarette counts for a randomized, placebo-controlled trial of bupropion with only TLFB measurements available. To account for repeated measurements on a subject, we induce correlation in the imputed counts. Finally, we analyze the imputed data in a longitudinal model that accommodates random subject effects and zero inflation. Both raw and imputed data show a significant drug effect for reducing the odds of non-abstinence and the number of cigarettes smoked among non-abstainers, but the imputed data provide efficiency gains. This method permits the analysis of daily cigarette consumption data previously deemed suspect due to reporting error and is applicable to other self-reported count data sets for which calibration samples are available. Copyright © 2015 John Wiley & Sons, Ltd.


DR.BEAT: First Insights into a Study to Collect Baseline BCG Data with a Sensor-Based Wearable Prototype in Heart-Healthy Adults.
The DR.BEAT project aims at the further development of a measurement system for recording ballistocardiographic signals into a body-worn sensor system combined with extensive signal processing, data evaluation and visualization. With a first breadboard prototype, an explorative feasibility study for acquiring initial signals of healthy cardiac activity in adults was performed. This paper briefly presents the DR.BEAT project, the breadboard prototype, the study conducted, and initial insights into the study results. The signals obtained in the study exhibit the seismocardiographic characteristics as reported in the literature and form the basis for further development of the hardware as well as the pre-processing and automated analysis algorithms in the DR.BEAT project.Clinical Relevance- The characteristics of ballisto- and seismocardiographic signals allow to infer about the mechanical work of the heart. The development of a body-worn sensor system to record ballisto- and seismocardiographic signals, compact enough for everyday wear, enables the acquisition of heart-specific parameters in terrestrial as well as extraterrestrial application scenarios. Combined with extensive signal analysis and visualization, it holds the potential to monitor heart health in a variety of contexts and support its maintenance and improvement.