Implantable selective stimulator to improve bladder voiding: design and chronic experiments in dogs.
Among the treatments to enhance the bladder voiding, the sacral roots neurostimulation is one of the most promising techniques. The electrostimulation of sacral nerves provokes a simultaneous contraction of the detrusor muscle as well as the external urethral sphincter (EUS). A new simplified-architecture implantable stimulator with its wireless controller have been designed to investigate high-frequency inhibition stimulation strategies. This innovative technique based on high-frequency inhibition reduces sphincter activity during stimulation. Low-frequency current pulses also applied to the sacral roots induces contraction of the detrusor muscle resulting in low pressure voiding. Chronic experiments were carried out on ten male mongrel paraplegic dogs. One cuff electrode was implanted along with each stimulator for eight months. The animals were stimulated twice a day using the prototypes of our implantable selective stimulator while voided and residual urine volume were measured during the procedure. These experiments revealed that the proposed stimulation strategy enhances bladder voiding by more than 50% in comparison with low-frequency only stimulation. The residual urine volume was reduced to an average of 9% and low pressure micturition was achieved as shown by weekly cystourethrogram.


Impaired skeletal muscle substrate oxidation in glucose-intolerant men improves after weight loss.
An impaired fatty acid handling in skeletal muscle may be involved in the development of insulin resistance and diabetes mellitus type 2 (DM2). We investigated muscle fatty acid metabolism in glucose-intolerant men (impaired glucose tolerance (IGT)), a prediabetic state, relative to BMI-matched control men (normal glucose tolerance (NGT)) during fasting and after a meal, because most people in the western society are in the fed state most of the day. Skeletal muscle free fatty acid (FFA) uptake and oxidation were studied using the stable isotope tracer [2,2-(2)H]-palmitate and muscle indirect calorimetry in the forearm model during fasting and after a mixed meal (33 energy % (E%) carbohydrates, 61 E% fat). Intramyocellular triglycerides (IMTGs) were monitored with 1H-magnetic resonance spectroscopy. IGT men were re-examined after weight loss (-15% of body weight (BW)). The postprandial increase in forearm muscle respiratory quotient (RQ) was blunted in IGT compared to NGT, but improved after weight loss. Weight loss also improved fasting-fat oxidation and tended to decrease IMTGs (P=0.08). No differences were found in fasting and postprandial forearm muscle fatty acid uptake between NGT and IGT, or in IGT before and after weight loss. The ability to switch from fat oxidation to carbohydrate oxidation after a meal is already impaired in the prediabetic state, suggesting this may be an early factor in the development toward DM2. This impaired ability to regulate fat oxidation during fasting and after a meal (impaired metabolic flexibility) can be (partly) reversed by weight loss.


Comparative growth of Malay, Chinese and Indian school children in Malaysia.
The weights and heights of 3,312 Malaysian primary school boys and girls, aged 6 to 11 years, belonging to various ethnic groups in Malaysia were measured. On the whole, the Chinese children were taller and heavier than the Malay and the Indian children who were the least heavy among the three ethnic groups. Economically the Indians were the poorest among the three ethnic groups and they also had the largest family size. When the household incomes were taken into consideration it was found that the growth achievement of the higher income children was better than that of the poorer children, irrespective of their ethnic groups. It is interesting to note that, although the Indian children as a whole, were the least heavy of the three ethnic groups, yet the growth achievement of the higher income Indian children was similar to that of the higher income Chinese children. The differences in growth achievement of the various ethnic groups are probably due to environmental differences, rather than genetic differences. It seems likely that Malaysian children of different ethnic groups (Malay, Chinese and Indian) can attain similar statures if environmental conditions are similar.


An asthma action plan created by physician, educator and patient online collaboration with usability and visual design optimization.
Asthma action plans (AAPs), which decrease hospitalizations and improve symptom control, are recommended in guidelines, but are seldom delivered to patients. Existing AAPs have been developed by experts, without the inclusion of all stakeholders (such as patients with asthma) and without specifically addressing usability and visual design. Our objective was to develop a more usable AAP by involving all stakeholders and considering design preferences. We created a Wiki-based system for multiuser AAP development. Pulmonologists, primary care physicians, asthma educators and patients used the system to collaboratively compile a single AAP by making multiple online selections over 1 week. We combined common elements from 3 AAPs developed in this way into 1, optimized visual design features and tested face validity in focus groups. A total of 41 participants averaged 646 selections/week over a login-time of 28.8 h/week. Of 35 participants, 28 (80%) were satisfied with the final AAP and 32 (91%) perceived that they would be able to use it. The plans created by the 3 groups were very similar, with a unanimous or majority agreement in the handling of 100/110 (91%) AAP options. Inclusion of multiple stakeholders and focus on design preferences predict enhanced usability and uptake of medical tools. The validity of our AAP is further supported by the similarity between the AAPs created by each group, user engagement and satisfaction with the plan and agreement with existing validity criteria proposed by experts. This AAP can be implemented in care with a concurrent measurement of uptake and health impact.


Dr. Allen Oldfather Whipple (1881-1963): Namesake of the pancreaticoduodenectomy.
Allen O. Whipple was an American surgeon who popularized the pancreaticoduodenectomy (Whipple procedure) for periampullary cancer, which remains the gold standard for pancreatic tumor resections. Whipple was educated at Princeton University (B.S., 1904) and Columbia University College of Physicians and Surgeons (M.D., 1908). He swiftly ascended the academic ranks, culminating in his appointment as Professor of Surgery at Columbia and Director of Surgical Services at Presbyterian Hospital in 1921. Whipple published three criteria (Whipple's triad) for evaluating hyperinsulinism secondary to pancreatic insulinoma. He also revived interest in portocaval anastomosis to reduce portal hypertension, determining it to be a consequence of liver disease. During his 40-year career, Whipple introduced the concept of multidisciplinary teams and prospective data collection. He also shaped the structure of surgical training as President of the American Surgical Association and Chairman of the American Board of Surgery. Beyond the walls of the operating room, Whipple was a Renaissance Man whose childhood in Persia (Iran) engendered a lifelong interest in the region's art, culture, history, and medicine. Dr. Allen Oldfather Whipple is remembered as a pioneering physician and surgeon beloved by those who trained under him.


Long-term effects of pneumatic dilatation on symptoms and lower oesophageal sphincter pressure in achalasia.
Pneumatic dilatation is an effective treatment for achalasia. Although follow-up studies have shown that its clinical benefit persists for years, it is still unknown whether the decrease in lower oesophageal sphincter (LOS) pressure is maintained over time. Our aim is to assess the long-term effects of pneumatic dilatation on symptoms and LOS pressure in patients with idiopathic achalasia. Eleven consecutive patients with achalasia (7 men) underwent I (n = 5) or 2 (n = 6) pneumatic dilatations (Rigiflex dilator) in order to achieve a stable (>1 year) clinical remission. Clinical scores (0-12, with scores of <3 indicating remission) and LOS pressure (sleeve manometry) were determined before treatment, after 3 and 12 months, and then every year for 6 years. No operative complications occurred. The patients showed a marked clinical improvement (2.0 (2.0-2.0), median (IQ range), after 3 months versus 8.0 (7.2-9.0) before treatment; P < 0.001), which was maintained throughout the follow-up period. Concurrently, there was a marked decrease in LOS pressure (5.0 (4.0-7.0) mmHg after 3 months versus 25.0 (20.0-36.2) before treatment; P < 0.001), which also remained stable over time. One or two pneumatic dilatations induce stable clinical remission and a decrease in LOS pressure that remains unchanged over time. Our data further support the use of dilatations as first-line treatment of achalasia.


A database of dispersion-induction DI, electrostatic ES, and hydrogen bonding α1 and β1 solvent parameters and some applications to the multiparameter correlation analysis of solvent effects.
For about 300 solvents, we propose a database of new solvent parameters describing empirically solute/solvent interactions: DI for dispersion and induction, ES for electrostatic interactions between permanent multipoles, α1 for solute Lewis base/solvent Lewis acid interactions, and β1 for solute hydrogen-bond donor/solvent hydrogen-bond acceptor interactions. The main advantage over previous parametrizations is the easiness of extension of this database to newly designed solvents, since only three probes, the betaine dye 30, 4-fluorophenol, and 4-fluoroanisole are required. These parameters can be entered into the linear solvation energy relationship A = A0 + di(DI) + eES + aα1 + bβ1 to predict a large number of varied physicochemical properties A and to rationalize the multiple intermolecular forces at the origin of solvent effects through a simple examination of the sign and magnitude of regression coefficients di, e, a, and b. Such a rationalization is illustrated for conformational and tautomeric equilibria and is supported by quantum-mechanical calculations.


Efficacy of regular sauna bathing for chronic tension-type headache: a randomized controlled study.
Chronic tension-type headache (CTTH) is a chronic syndrome characterized by frequent headache occurring more than 15 days per month. The intensity and duration of headache pain can be very distressing and disabling on an individuals' well-being. The purpose of this study was to examine the applicability of sauna bathing as a new method of treatment for reducing pain in patients with CTTH. Thirty-seven people who fulfilled the International Headache Society criteria for CTTH were randomly assigned into two groups. The control group (n=20) received advice and education while the intervention group (n=17) received the same advice and attended a sauna regularly for 8 weeks. Reductions in subjective pain were determined using the numerical pain rating scale (NPRS). Disturbance in sleep, depression as assessed by Beckman Disability Index (BDI), and Headache Disability Index (HDI) were measured. Mean change in headache intensity significantly differed between the sauna and control group by 1.27 (95% confidence interval, 0.48-2.07; F=10.17; df=1, 117; p=0.002). There was no statistically significant change in duration of headache or improvement in sleep, depression, or HDI between the sauna and control groups. Regular sauna bathing is a simple, self-directed treatment that is effective for reducing headache pain intensity in CTTH.


Third-Generation Sequencing in the Clinical Laboratory: Exploring the Advantages and Challenges of Nanopore Sequencing.
Metagenomic sequencing for infectious disease diagnostics is an important tool that holds promise for use in the clinical laboratory. Challenges for implementation so far include high cost, the length of time to results, and the need for technical and bioinformatics expertise. However, the recent technological innovation of nanopore sequencing from Oxford Nanopore Technologies (ONT) has the potential to address these challenges. ONT sequencing is an attractive platform for clinical laboratories to adopt due to its low cost, rapid turnaround time, and user-friendly bioinformatics pipelines. However, this method still faces the problem of base-calling accuracy compared to other platforms. This review highlights the general challenges of pathogen detection in clinical specimens by metagenomic sequencing, the advantages and disadvantages of the ONT platform, and how research to date supports the potential future use of nanopore sequencing in infectious disease diagnostics.


The 'Natural' Accord of DuBois and Washington: An Environmentally Racialized Consciousness.
The conflict and discord between Booker T. Washington and W.E.B DuBois regarding their premise and approach to racial uplift for Black Americans have been very well documented. While Washington sought equality with accommodation, DuBois functioned through agitation. However, their biophilic accord and unity within the natural environment have been both underrecognized and underappreciated. As an honor to these esteemed racial and social justice giants, this special issue article reveals the universality of their environmental justice ideologies while also celebrating the beauty, power and foresight of their ecological language in script and speech.


Membrane separations for solid-liquid clarification within lignocellulosic biorefining processes.
Membrane separations can be integrated into a biorefinery to reduce water and energy consumption. Unfortunately, current membrane materials suffer from severe fouling, which limits their applicability. Here, using analytical characterizations along with fouling models, we correlate membrane properties with performance metrics to provide a framework for optimal membrane selection during solid-liquid clarification of a biomass hydrolysate. Five membranes were evaluated: polyether sulfone, mixed cellulose esters, and three surface modified membranes with weak acid, strong acid, and weak base functionalities. Lignin was the primary component responsible for flux decline, due to physical entrapment and chemical adsorption. The best membrane performance (high and sustained flux, low fouling, and high separation factor) was correlated with higher surface roughness, lower hydrophobicity, neutral or positively charged zeta potential, and a larger number of smaller surface pores. These analyses provide valuable information for designing new materials for biorefining processes to reduce fouling and increase stability.


Investigation of time series-based genre popularity features for box office success prediction.
Predicting the profitability of movies at the early phase of production can be helpful to support the decision to invest in movies however, due to the limited information at this stage it is a challenging task to predict the movie's profitability. This study proposes genre popularity features using time series prediction. We argue that a movie can produce better box office returns if its genre's popularity is high at the time of release. The novel genre popularity features are proposed in terms of budget, revenue, frequency, success, and return on investment (ROI). The proposed features couple the predicted genre popularity with release time, in order to train the machine learning classifiers. The experimentation shows that the Gradient Boosting classifier gained a significant improvement using proposed features and achieved an accuracy of more than 92.4%, i.e., 35.7% better than an existing state of the art study considering a multi-class problem.


Blood tracer kinetics in the arterial tree.
Evaluation of blood supply of different organs relies on labeling blood with a suitable tracer. The tracer kinetics is linear: Tracer concentration at an observation site is a linear response to an input somewhere upstream the arterial flow. The corresponding impulse response functions are currently treated empirically without incorporating the relation to the vascular morphology of an organ. In this work we address this relation for the first time. We demonstrate that the form of the response function in the entire arterial tree is reduced to that of individual vessel segments under approximation of good blood mixing at vessel bifurcations. The resulting expression simplifies significantly when the geometric scaling of the vascular tree is taken into account. This suggests a new way to access the vascular morphology in vivo using experimentally determined response functions. However, it is an ill-posed inverse problem as demonstrated by an example using measured arterial spin labeling in large brain arteries. We further analyze transport in individual vessel segments and demonstrate that experimentally accessible tracer concentration in vessel segments depends on the measurement principle. Explicit expressions for the response functions are obtained for the major middle part of the arterial tree in which the blood flow in individual vessel segments can be treated as laminar. When applied to the analysis of regional cerebral blood flow measurements for which the necessary arterial input is evaluated in the carotid arteries, present theory predicts about 20% underestimation, which is in agreement with recent experimental data.


Non-invasive ventilation after cardiac surgery outside the Intensive Care Unit.
Non-invasive ventilation (NIV) can prevent or treat postoperative acute respiratory failure. NIV after discharge from the Intensive Care Unit (ICU) has never been described in the setting of cardiac surgery. This study enrolled 85 patients who received NIV in the main ward as treatment for respiratory failure. The patients had the following conditions: atelectasis (45 patients), pleural effusion (20 patients), pulmonary congestion (13 patients), diaphragm hemiparesis (6 patients), pneumonia (4 patients) or a combination of these conditions. Eighty-three patients were discharged from the hospital in good condition and without need for further NIV treatment, while two died in-hospital. Four of the 85 patients had an immediate NIV failure, while eight patients had delayed NIV failure. Only one patient had a NIV-related complication represented by hypotension after NIV institution. In this patient, NIV was interrupted with no consequences. Major mistakes were mask malpositioning with excessive air leaks (7 patients), incorrect preparation of the circuit (one patient), and oxygen tube disconnection (one patient). Minor mistakes (sub-optimal positioning of the face mask without excessive air leaks) were noted by the respiratory therapists for all patients and were managed by slightly modifying the mask position. In our experience, postoperative NIV is feasible, safe and effective in treating postoperative acute respiratory failure when applied in the cardiac surgical ward, preserving intensive care unit beds for surgical activity. A respiratory therapy service managed the treatment in conjunction with ward nurses, while an anesthesiologist and a cardiologist served as consultants.


Knowledge gaps and research priorities in Staphylococcus aureus mastitis control.
This study assessed knowledge gaps and suggested research priorities in the field of Staphylococcus aureus mastitis. Staphylococcus aureus infecting the mammary gland remains a major problem to the dairy industry worldwide because of its pathogenicity, contagiousness, persistence in the cow environment, colonization of skin or mucosal epithelia, and the poor curing efficacy of treatments. Staphylococcus aureus also constitutes a threat to public health due to food safety and antibiotic usage issues and the potential for bidirectional transmission of strains between humans and dairy animals (cows and small ruminants). Gaps have been identified in (i) understanding the molecular basis for pathogenesis of S. aureus mastitis, (ii) identifying staphylococcal antigens inducing protection and (iii) determining the cell-mediated immune responses to infection and vaccination. The recommended priorities for research are (i) improved diagnostic methods for early detection of infection and intervention through treatment or management, (ii) development of experimental models to investigate the strategies used by S. aureus to survive within the mammary gland and resist treatment with anti-microbials, (iii) investigation of the basis for cow-to-cow variation in response to S. aureus mastitis, (iv) identification of the immune responses (adaptive and innate) induced by infection or vaccination and (v) antibacterial discovery programmes to develop new, more effective, narrow spectrum antibacterial agents for the treatment of S. aureus mastitis. With the availability and ongoing improvement of molecular research tools, these objectives may not be out of reach in the future.


Oncogenic pathways that affect antitumor immune response and immune checkpoint blockade therapy.
Mechanistic insights of cancer immunology have led to the development of immune checkpoint blockade therapy (ICBT), which has elicited a remarkable clinical response in some cancer patients. Increasing evidence suggests that activation of oncogenic pathways, such as RAS/RAF/MAPK and PI3K signaling, impairs the antitumor immune response. Such oncogenic signaling, in turn, activates many inhibitory factors, including expression of immune checkpoint genes-allowing active infiltration of immunosuppressive cells into the tumor environment and inducing resistance against T-cell killing. In preclinical tumor models, effective targeting of oncogenic pathways has enhanced the response to ICBT. Ongoing clinical trials are now evaluating combination therapy (i.e., the use of oncogenic pathway inhibitors in combination with ICBT). However, more translational and clinical research is needed, to optimize ICBT doses and sequence, minimize toxicity, and assess the impact on study participants of certain genetic backgrounds. Also, it is crucial to understand whether wild-type tumors with elevated oncogenic signaling will respond to combination therapy. Insights gained through current and future translational studies will provide the scientific premise and rationale to target 1 or more oncogenic pathways in ICBT-resistant tumors, thus enabling more human patients to benefit from combination therapy.


Lability of host-cell DNA in growing cell cultures due to Mycoplasma.
In HeLa-cell cultures chronically infected with Mycoplasma, (PPLO) host-cell DNA is unstable as detected by incorporation of H(3)-or C1 i-thymidine into DNA and subsequent release into the medium as acidsoluble radioactivity. This characteristic can be transmitted to PPLO-free cultures of strain L cells by inoculation with preparations of PPLO from the HeLa cells, although chronically infected cultures of L cells continue to multiply. In addition, virus preparations also may carry PPLO contamination through numerous passages.


Arthroscopic single-bundle anterior cruciate ligament reconstruction with periosteum-enveloping hamstring tendon graft: clinical outcome at 2 to 7 years.
In this case-series outcome study, we present our surgical technique for single-bundle anterior cruciate ligament (ACL) reconstruction with periosteum-enveloping hamstring tendon graft at a minimum of 2 years' follow-up. From 2000 to 2005, ACL reconstruction with a periosteum-enveloping hamstring tendon graft was performed in 368 patients (372 knees). Of those patients, 312 who completed at least 2 years of follow-up were included for analysis. Four-strand periosteum-enveloping hamstring tendon grafts were used for single-bundle reconstruction. Clinical assessments included the Lysholm knee score, International Knee Documentation Committee score, KT-1000 instrumented testing (MEDmetric, San Diego, CA), thigh muscle assessment, and radiographic evaluation. Radiographs were used to assess femoral and tibial tunnel widening. The 312 study patients were followed up for a mean of 4.6 years (range, 2 to 7 years). The median Lysholm knee scores were 56 points (range, 40 to 70 points) and 95 points (range, 60 to 100 points) before and after surgery, respectively. After reconstruction, 85% of patients could return to moderate or strenuous activity, 5.1% exhibited grade 2 or higher ligament laxity with the anterior drawer test, and 6.1% had a positive pivot shift. Complete range of motion was achieved in 88% of patients. On the basis of International Knee Documentation Committee assessment, 93% of patients had a normal or nearly normal rating. Satisfactory results can be achieved with the periosteum-enveloping hamstring tendon graft in single-bundle ACL reconstruction with minimal tunnel widening. Bone tunnel enlargement of more than 1 mm was identified in 5.4% of femoral tunnels and 6.1% of tibial tunnels, which was less than in other studies using comparable fixation. Level IV, therapeutic case series.


Ergonomics observation: Harvesting tasks at oil palm plantation.
Production agriculture is commonly associated with high prevalence of ergonomic injuries, particularly during intensive manual labor and during harvesting. This paper intends to briefly describe an overview of oil palm plantation management highlighting the ergonomics problem each of the breakdown task analysis. Although cross-sectional field visits were conducted in the current study, insight into past and present occupational safety and health concerns particularly regarding the ergonomics of oil palm plantations was further exploited. Besides discussion, video recordings were extensively used for ergonomics analysis. The unique commodity of oil palm plantations presents significantly different ergonomics risk factors for fresh fruit bunch (FFB) cutters during different stages of harvesting. Although the ergonomics risk factors remain the same for FFB collectors, the intensity of manual lifting increases significantly with the age of the oil palm trees-weight of FFB. There is urgent need to establish surveillance in order to determine the current prevalence of ergonomic injuries. Thereafter, ergonomics interventions that are holistic and comprehensive should be conducted and evaluated for their efficacy using approaches that are integrated, participatory and cost-effective.


Efficient preparation of size tunable PEGylated gold nanoparticles.
A facile, one-step self-assembly of polymer nanoreactors is reported for the fabrication of uniform PEGylated gold nanoparticles. Nanoreactor assembly occurs within milliseconds and gold nanoparticles are produced within minutes at room temperature. The presented approach enables continuous synthesis of size tunable gold nanoparticles with tailorable surface chemistry.


Vitamin C antagonizes the cytotoxic effects of antineoplastic drugs.
Vitamin C is an antioxidant vitamin that has been hypothesized to antagonize the effects of reactive oxygen species-generating antineoplastic drugs. The therapeutic efficacy of the widely used antineoplastic drugs doxorubicin, cisplatin, vincristine, methotrexate, and imatinib were compared in leukemia (K562) and lymphoma (RL) cell lines with and without pretreatment with dehydroascorbic acid, the commonly transported form of vitamin C. The effect of vitamin C on viability, clonogenicity, apoptosis, P-glycoprotein, reactive oxygen species (ROS), and mitochondrial membrane potential was determined. Pretreatment with vitamin C caused a dose-dependent attenuation of cytotoxicity, as measured by trypan blue exclusion and colony formation after treatment with all antineoplastic agents tested. Vitamin C given before doxorubicin treatment led to a substantial reduction of therapeutic efficacy in mice with RL cell-derived xenogeneic tumors. Vitamin C treatment led to a dose-dependent decrease in apoptosis in cells treated with the antineoplastic agents that was not due to up-regulation of P-glycoprotein or vitamin C retention modulated by antineoplastics. Vitamin C had only modest effects on intracellular ROS and a more general cytoprotective profile than N-acetylcysteine, suggesting a mechanism of action that is not mediated by ROS. All antineoplastic agents tested caused mitochondrial membrane depolarization that was inhibited by vitamin C. These findings indicate that vitamin C given before mechanistically dissimilar antineoplastic agents antagonizes therapeutic efficacy in a model of human hematopoietic cancers by preserving mitochondrial membrane potential. These results support the hypothesis that vitamin C supplementation during cancer treatment may detrimentally affect therapeutic response.


Influence of Landmarks on Spatial Memory in Short-nosed Fruit Bat, Cynopterus sphinx.
In order to study the relationship between landmarks and spatial memory in short-nosed fruit bat, Cynopterus sphinx (Megachiroptera, Pteropodidae), we simulated a foraging environment in the laboratory. Different landmarks were placed to gauge the spatial memory of C. sphinx. We changed the number of landmarks every day with 0 landmarks again on the fifth day (from 0, 2, 4, 8 to 0). Individuals from the control group were exposed to the identical artificial foraging environment, but without landmarks. The results indicated that there was significant correlation between the time of the first foraging and the experimental days in both groups (Pearson Correlation: experimental group: r=-0.593, P<0.01; control group: r=-0.581, P<0.01). There was no significant correlation between the success rates of foraging and the experimental days in experimental groups (Pearson Correlation: r=0.177, P>0.05), but there was significant correlation between the success rates of foraging and the experimental days in the control groups (Pearson Correlation: r=0.445, P<0.05). There was no significant difference for the first foraging time between experimental and control groups (GLM: F(0.05,1 )=4.703, P>0.05); also, there was no significant difference in success rates of foraging between these two groups (GLM: F(0.05,1 )=0.849, P>0.05). The results of our experiment suggest that spatial memory in C. sphinx was formed gradually and that the placed landmarks appeared to have no discernable effects on the memory of the foraging space.


Evaluating the role of conserved amino acids in bacterial O-oligosaccharyltransferases by in vivo, in vitro and limited proteolysis assays.
Bacterial O-Oligosaccharyltransferases (O-OTases) constitute a growing family of enzymes that catalyze the transfer of a glycan from a lipid carrier to protein acceptors. O-OTases are inner membrane proteins that display limited sequence similarity, except for the Wzy_C signature domain also present in a predicted periplasmic loop of the WaaL ligase, the enzyme responsible for transferring the O antigen to the lipid A core. The mechanism of O-OTase-dependent glycosylation is poorly understood. In this work, conserved amino acid residues in the O-OTases were replaced with alanine in PglL, the O-OTase of Neisseria meningitidis. The activity of wild-type PglL and its mutant derivatives were analyzed in vivo in engineered Escherichia coli cells, and in in vitro assays. We identified two additional sites of pilin glycosylated exclusively by PglL in E. coli. Both sites are modified with phosphoglycerol (PG) by different enzymes in Neisseria gonorrhoeae and Neisseria meningitidis. Limited proteolysis experiments revealed a conformational change that is triggered upon interaction of the C-terminal region of PglL with the lipid-linked oligosaccharide (LLO) substrate. These experiments showed that Q178 and Y405 are required for optimal function, whereas H349 is essential for activity and plays a critical role in the interaction with LLO. The equivalent His residue is also essential for WaaL activity, which suggests a common mechanism for both enzymes, and supports the hypothesis that O-glycosylation and lipopolysaccharide (LPS) synthesis are evolutionarily related. These results contribute to the elucidation of the mechanism of O-OTases, which are promising targets for novel antibiotics and present an enormous potential for glycoengineering novel vaccines and therapeutics.


Thymocyte differentiation in gamma-irradiated severe-combined immunodeficient mice: characterization of intermediates and products of V(D)J recombination at the T cell receptor alpha locus.
Treatment with DNA-damaging agents promotes rescue of V(D)J recombination, limited thymocyte differentiation, and development of thymic lymphomas in severe-combined immunodeficient (SCID) mice. One intriguing aspect of this system is that irradiation rescues rearrangements at the T cell receptor (TCR) beta, gamma and delta loci, but not at the TCR alpha locus. Current models posit that only those loci that are recombinationally active at the time of irradiation can be rescued. Here, we employ sensitive, semiquantitative ligation-mediated polymerase chain reaction assays to detect a specific class of recombination intermediates, hairpin coding ends, at the TCR alpha locus. We found that J alpha-coding ends are undetectable in unirradiated SCID thymocytes, but accumulate after irradiation at times coincident with the emergence of a CD4+ CD8+ thymocyte population. Coding joints produced by joining of these ends, however, are extremely rare. To test whether the presence of hairpin coding ends at TCR alpha is sufficient for irradiation-mediated rescue of coding joint formation, we administered a second dose of gamma-irradiation after abundant CD4+ CD8+ thymocytes and hairpin TCR alpha coding ends had accumulated. This treatment failed to stimulate rescue of TCR alpha coding joints. Thus, the presence of hairpin coding ends at the time of irradiation, while perhaps necessary, is not sufficient for rescue of V(D)J rearrangements. These results support a refined model for irradiation-mediated rescue of TCR rearrangements in SCID mice.


Fibroblast Promotes Head and Neck Squamous Cell Carcinoma Cell Invasion through Mechanical Barriers in 3D Collagen Microenvironments.
Cancer metastasis involves not only cancer cells but also fibroblasts and the surrounding collagen matrices. Previous studies have reported that in tumor tissues, cancer cells and fibroblasts surrounded by dense collagen are often associated with a high risk of cancer metastasis. However, the mechanism of the interaction between the cancer cells, fibroblasts, and the surrounding collagen matrices in vivo to promote cancer cell invasion in different collagen concentration environments remains unclear. To address this issue, we cocultured head and neck squamous cell carcinoma (OECM-1 cells) and human dermal fibroblasts (HDFs) to form 3D spheroids, embedded in collagen gel with different concentrations to delineate their roles and their interactions in cancer cell invasion. We showed that in single-species spheroids, the OECM-1 cells could not remodel the high-concentration (8 mg/mL) collagen matrices to invade into the surrounding collagen. In contrast, in the coculture spheroids, the HDF cells could remodel the collagen matrices, via MMP-meditated collagen degradation, to increase the invasion capability of OECM-1 cells. In the case of low-concentration (2 mg/mL) collagen matrices, both HDF and OECM-1 cells in the coculture spheroids could independently invade into the surrounding collagen via force remodeling of collagen. Our results revealed that the assistance of HDFs was critical for OECM-1 cell invasion into the surrounding extracellular matrix with high collagen concentration, high storage modulus, and small pore sizes. These insightful results shed light on the possible optimal invasion strategy of cancer tumors in vivo in response to different storage moduli of surrounding collagen matrices.


Dietary Inflammatory Index and Clinical Outcome Measures in Adults With Moderate-to-Severe Asthma.
Diet is increasingly recognized as a modifiable factor in lung health, predominantly due to the immunomodulatory effects of nutrients. The Dietary Inflammatory Index (DII) is a score developed to express the inflammatory potential of a diet. We aimed to assess the association of the DII and food groups, with clinical, functional, and inflammatory asthma outcomes in adults with asthma. Patients with moderate-to-severe asthma were included in this cross-sectional study between June 2019 and October 2021, and completed a 3-day food diary, to calculate the DII and intake of food groups (ie, fruits, whole grains, processed meats, and sugar-sweetened beverages). Functional outcomes included pulmonary function tests and the 6-minute walking distance, whereas clinical outcomes were assessed using questionnaires on asthma control, quality of life, and health care utilization. Inflammatory markers were exhaled nitric oxide and blood leukocytes, eosinophils, and IL-6. Multivariable regression analyses were used to examine the association of DII and food groups with asthma outcomes. A total of 109 patients participated (35% male, mean ± standard deviation age 51.8 ± 14.2 years, body mass index 27.4 ± 5.3 kg/m2). Overall, 62% had a DII score >0, indicating a proinflammatory diet, which was not related to asthma severity. A more proinflammatory diet was consistently associated with lower forced vital capacity (%pred), but inconsistent results were observed with respect to airway obstruction. Neither the DII nor food groups were associated with clinical outcomes. Except for higher levels of exhaled nitric oxide in relation to an anti-inflammatory diet, we found no associations between inflammatory markers and the DII. Results from this cross-sectional study among patients with moderate-to-severe asthma do not support the hypothesis that a proinflammatory diet is associated with worse asthma outcomes, although limitations in study design and dietary intake estimation should be considered. Future well-designed experimental studies are needed to assess whether targeting the inflammatory potential of diet could lead to better outcomes in adults with asthma.


The changing preference of T and B cells for partners as T-dependent antibody responses develop.
Recirculating virgin CD4+ T cells spend their life migrating between the T zones of secondary lymphoid tissues where they screen the surface of interdigitating dendritic cells. T-cell priming starts when processed peptides or superantigen associated with class II MHC molecules are recognised. Those primed T cells that remain within the lymphoid tissue move to the outer T zone, where they interact with B cells that have taken up and processed antigen. Cognate interaction between these cells initiates immunoglobulin (Ig) class switch-recombination and proliferation of both B and T cells; much of this growth occurs outside the T zones B cells migrate to follicles, where they form germinal centres, and to extrafollicular sites of B-cell growth, where they differentiate into mainly short-lived plasma cells. T cells do not move to the extrafollicular foci, but to the follicles; there they proliferate and are subsequently involved in the selection of B cells that have mutated their Ig variable-region genes. During primary antibody responses T-cell proliferation in follicles produces many times the peak number of T cells found in that site: a substantial proportion of the CD4+ memory T-cell pool may originate from growth in follicles.


Patient-to-nurse ratios: Balancing quality, nurse turnover, and cost.
We consider the problem of setting appropriate patient-to-nurse ratios in a hospital, an issue that is both complex and widely debated. There has been only limited effort to take advantage of the extensive empirical results from the medical literature to help construct analytical decision models for developing upper limits on patient-to-nurse ratios that are more patient- and nurse-oriented. For example, empirical studies have shown that each additional patient assigned per nurse in a hospital is associated with increases in mortality rates, length-of-stay, and nurse burnout. Failure to consider these effects leads to disregarded potential cost savings resulting from providing higher quality of care and fewer nurse turnovers. Thus, we present a nurse staffing model that incorporates patient length-of-stay, nurse turnover, and costs related to patient-to-nurse ratios. We present results based on data collected from three participating hospitals, the American Hospital Association (AHA), and the California Office of Statewide Health Planning and Development (OSHPD). By incorporating patient and nurse outcomes, we show that lower patient-to-nurse ratios can potentially provide hospitals with financial benefits in addition to improving the quality of care. Furthermore, our results show that higher policy patient-to-nurse ratio upper limits may not be as harmful in smaller hospitals, but lower policy patient-to-nurse ratios may be necessary for larger hospitals. These results suggest that a "one ratio fits all" patient-to-nurse ratio is not optimal. A preferable policy would be to allow the ratio to be hospital-dependent.


Rehabilitation and COVID-19: a rapid living systematic review 2020 by Cochrane Rehabilitation Field. Update as of October 31st, 2020.
This living systematic review presents the monthly update of the second edition of the rapid living systematic review 2020 conducted by Cochrane Rehabilitation REH-COVER Action Steering Committee. The aim of this study was to update the monthly COVID-19 and rehabilitation literature research up to October 31st, 2020. Methodology described in the second edition of the rapid living systematic review 2020 conducted by Cochrane Rehabilitation REH-COVER action was applied. PubMed, Embase, CINAHL, Scopus, Web of Science, and PEDro databases were searched, and papers related to COVID-19 and rehabilitation were retrieved and summarized descriptively. The database search retrieved 2704 publications. Duplicates were removed, and 1185 unique records were screened for inclusion. After screening titles, abstracts and full-texts, 22 papers were included in the present review. According to OCEBM 2011 Levels of Evidence table, 17 studies (77%) fall within the level of evidence 4 category, while the remainder (23%) are categorized as level of evidence 3. Most studies (N.=19; 86%) provided epidemiological data about the disease natural history/determining factor or the clinical presentation of COVID-19 infection, while only two studies focused on health service organization and intervention efficacy. The most recent published COVID-19 research relevant to rehabilitation primarily provides data on the clinical course and the clinical presentation of the pathology, rather than on rehabilitation interventions or service delivery. Studies with high levels of evidence regarding the efficacy of interventions, long-term monitoring, or new health service organization models are lacking.


Does vasopressin sustain blood pressure of normally hydrated healthy volunteers?
The inhibitor of the pressor effect of arginine vasopressin (AVP), d(CH2)5Tyr(Me)AVP, at a dose of 5 micrograms/kg iv was shown in four healthy volunteers to antagonize the blood pressure, heart rate, and skin blood flow response to a lysine vasopressin infusion of 1 mIU X kg-1 X min-1. The inhibition lasted for more than 2 h. When the same dose of the vasopressin antagonist was administered to 10 healthy normally hydrated volunteers with their renin system intact or acutely blocked by 25 mg of captopril po, none of the above parameters changed. It is concluded that circulating vasopressin, even in the face of a blocked renin-angiotensin system, does not actively contribute to maintenance of cardiovascular homeostasis.


Designer receptors: therapeutic adjuncts to cell replacement therapy in Parkinson's disease.
Cell replacement for restoring neuronal populations in Parkinson's disease has been demonstrated as a potential therapeutic strategy over several decades of studies; however, a number of issues regarding sources of replacement neurons and optimization of therapeutic efficacy in vivo have hampered clinical implementation. In this issue of the JCI, Dell'Anno and colleagues evaluated the use of induced dopaminergic (iDA) neurons that were generated by direct fibroblast reprogramming for transplantation and demonstrated that postmitotic iDA neurons stably and functionally integrate into host striatum to produce motor improvements in 6-OHDA rats, a Parkinson's disease model. Furthermore, using designer receptors exclusively activated by designer drugs (DREADDs) in iDA grafts to noninvasively increase dopamine release from grafted neurons, the authors were able to remotely control transplanted neurons and enhance therapeutic efficacy. This initial proof-of-concept study is the first application of DREADD technology to treat neurodegenerative dysfunction, and by using DREADDs as an adjunct to iDA cell therapy, it presents a novel strategy to overcome some current caveats of cell replacement therapy.


A peroxidase gene family and gene trees in Heterobasidion and related genera.
Four putative peroxidase-encoding gene fragments, named mnp1a, mnp1b, mnp2 and mnp3, were amplified with degenerative primers from the white-rot basidiomycete genus Heterobasidion. The fragments were cloned and sequenced. Similar fragments were produced and analyzed from the related genera Amylostereum, Bondarzewia and Echinodontium. Each amplified fragment contains three identically positioned introns. According to the predicted amino acid sequence, these fragments are most similar to two Mn peroxidase-encoding genes (MPGI and mnp2) and gene pgv of Trametes versicolor. Conserved residues thought to be essential for peroxidase function were identified. All four MnP gene loci of Heterobasidion were detected only in H. parviporum. Variation occurred in the predicted amino-acid sequences (131-132 amino acids) of all four fragments originating from the 47 Heterobasidion isolates tested. Amino acid variation in fragments of mnp2 and mnp3 separated European Heterobasidion parviporum ("S-type") and H. abietinum ("F-type"), known to have identical rDNA sequences. Asian and western North American isolates from fir, spruce and other hosts had the peroxidase amino acid sequences of European H. parviporum. American and European H. annosum ("P-type") isolates had different amino acid sequences and might be cryptic species.


Molecular alignment-assisted spectral broadening and shifting in the near-infrared with a recycled depleted pump from an optical parametric amplifier.
We demonstrate how the depleted pump of an optical parametric amplifier can be recycled for impulsive alignment of a molecular gas inside a hollow-core fiber and use such alignment for the broadening and frequency shift of the signal pulse at a center wavelength of ∼1300 nm. Our results combine non-adiabatic molecular alignment, self-phase modulation, and Raman non-linearities. We demonstrate spectral shifts of up to 204 nm and a spectral broadening of more than one octave. We also report on the time delays at which broadening occurs, which do not coincide with any of the molecular rotational constants. Further, we encounter that maximum frequency shifts occur when the signal and pump have perpendicular polarization instead of parallel.


Development of estimation methods of skin oxidation and evaluation of anti-oxidative effects of genistein in topical formulations.
The objective of the present study was to establish the method of measurement of hydrogen peroxide and to estimate the anti-oxidative effect of genistein in the skin. UVB induced skin oxidation and anti-oxidative effect of genistein formulations were evaluated by determining levels of hydrogen peroxide. The mechanism involved in the determination of hydrogen peroxide is based on a color reaction between ferric ion (Fe(3+)) and xylenol orange, often called FOX assay and subsequent monitoring of absorbance values of the reactant at 540 nm. The reaction was to some extent pH-dependent and detection sensitivity was greatest at pH 1.75. Genistein liposomal gel demonstrated better anti-oxidative effect with regard to lowering hydrogen peroxide levels elevated by UVB irradiation compared to genistein-suspended gel. A linear relationship has been observed between anti-oxidative effect of genistein and drug deposition in the skin tissue. Genistein liposomal gel resulting in the localization of the drug in the deeper skin led to improved anti-oxidative effect compared to genistein gel. The suggested method for evaluation of oxidation of the skin can be used as a tool to screen effective anti-oxidative agents and their delivery systems acting on the skin.


Risking one's life to save one's livelihood: Precarious work, presenteeism, and worry about disease exposure during the COVID-19 pandemic.
The present study advances research on the negative consequences of precarious work experiences (PWE), which include perceptions of threats to one's job and financial security as well as a sense of powerlessness and inability to exercise rights in the workplace. Using the COVID-19 pandemic as a backdrop, we examine how PWE relate to sickness presenteeism and worry about work-related COVID-19 exposure. In a 12-week, four-wave study of workers working fully in-person, perceptions of powerlessness and job insecurity were associated with presenteeism (e.g., general presenteeism as well as attending work with known or possible COVID-19 infection) and concerns about disease exposure at work. Whereas powerlessness primarily operated at the between-person level of analysis, job insecurity's effects emerged at both levels of analysis. A sense of powerlessness at work also predicted sending children to school/daycare sick. In sum, the findings suggest that precarity related to being able to keep one's job and a sense of powerlessness at work contribute to concerns about the risk of COVID-19 exposure at work and, simultaneously, behaviors that may contribute to the health risks faced by others. This research provides added support to the argument that precarious work should be addressed in order to improve both worker well-being and public health. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


Intermittent hypoxia inhibits clearance of triglyceride-rich lipoproteins and inactivates adipose lipoprotein lipase in a mouse model of sleep apnoea.
Delayed lipoprotein clearance is associated with atherosclerosis. This study examined whether chronic intermittent hypoxia (CIH), a hallmark of obstructive sleep apnoea (OSA), can lead to hyperlipidaemia by inhibiting clearance of triglyceride rich lipoproteins (TRLP). Male C57BL/6J mice on high-cholesterol diet were exposed to 4 weeks of CIH or chronic intermittent air (control). FIO(2) was decreased to 6.5% once per minute during the 12 h light phase in the CIH group. After the exposure, we measured fasting lipid profile. TRLP clearance was assessed by oral gavage of retinyl palmitate followed by serum retinyl esters (REs) measurements at 0, 1, 2, 4, 10, and 24 h. Activity of lipoprotein lipase (LpL), a key enzyme of lipoprotein clearance, and levels of angiopoietin-like protein 4 (Angptl4), a potent inhibitor of the LpL activity, were determined in the epididymal fat pads, skeletal muscles, and heart. Chronic intermittent hypoxia induced significant increases in levels of total cholesterol and triglycerides, which occurred in TRLP and LDL fractions (P< 0.05 for each comparison). Compared with control mice, animals exposed to CIH showed increases in REs throughout first 10 h after oral gavage of retinyl palmitate (P< 0.05), indicating that CIH inhibited TRLP clearance. CIH induced a >5-fold decrease in LpL activity (P< 0.01) and an 80% increase in Angptl4 mRNA and protein levels in the epididymal fat, but not in the skeletal muscle or heart. CIH decreases TRLP clearance and inhibits LpL activity in adipose tissue, which may contribute to atherogenesis observed in OSA.


Adjusted P values for genome-wide scans.
Genome-wide scans for quantitative trait loci (QTL) have traditionally been summarized with plots of logarithm of odds (LOD) scores. A valuable modification is to supplement such plots with an additional vertical axis displaying quantiles of adjusted P values and labeling local maxima of the LOD scores with location-specific adjusted P values. This provides a visible gradation of genome-wide significance for the LOD score curve, instead of the stark dichotomy that a single threshold yields. Adjusted P values give genome-wide significance of individual LOD scores and are obtained through a straightforward modification of the familiar algorithm for generating permutation-based thresholds.


Why currently used diagnostic techniques for heart failure in rheumatoid arthritis are not enough: the challenge of cardiovascular magnetic resonance imaging.
Rheumatoid arthritis (RA) is a multiorgan inflammatory disorder affecting approximately 1% of the population that leads to progressive joint destruction and disability. Patients with RA exhibit a high risk of cardiovascular disease, which results in premature morbidity and mortality and reduced life expectancy, when compared with the general population. Among various guises of myocardial involvement, heart failure (HF) has been recently recognized as an important contributory factor to the excess cardiovascular mortality associated with RA. HF in RA typically presents with occult clinical symptomatology and is mainly associated with structural and functional left ventricular abnormalities leading to diastolic dysfunction, while systolic myocardial performance remains well preserved. As isolated diastolic dysfunction is a predictor of high mortality, the evaluation of patients in early asymptomatic stages, when treatment targeting the heart is more likely to be effective, is of great importance. Although patient history and physical examination remain the cornerstones of HF evaluation, noninvasive imaging of cardiac chambers, coronary arteries, and great vessels may be necessary. Echocardiography, nuclear techniques, and invasive coronary angiography are already established in the routine assessment of HF; however, many aspects of HF pathophysiology in RA remain obscure, due to the limitations of currently used techniques. The capability of cardiovascular magnetic resonance (CMR) to capture early tissue changes allows timely detection of pathophysiologic phenomena of HF in RA, such as myocardial inflammation and myocardial perfusion defects, due to either macrovascular (coronary artery disease) or microvascular (vasculitis) disease. Therefore, CMR may be a useful tool for early, accurate diagnosis and research in patients with RA.


FOLFIRINOX in Pancreatic Cancer: Risk Factors for Febrile Neutropenia and Severe Neutropenia - Nationwide Study Analysis.
FOLFIRINOX (FFX) is a standard treatment for patients with advanced pancreatic cancer. However, it often causes serious hematological adverse events. This study aimed to identify the risk factors for febrile neutropenia (FN) and grade 4 (G4) neutropenia during treatment with FFX in the real world. We analyzed data obtained from a nationwide multicenter observational study (JASPAC 06) that included 399 patients with unresectable or recurrent pancreatic cancer who received FFX at 27 institutions in Japan. Nadir neutrophil counts occurred from day 8 to day 22 of cycle 1, and granulocyte colony-stimulating factor was administered to over a quarter of the patients in the first cycle. Of 399 patients, FN and G4 neutropenia occurred in 51 (13%) and 108 (27%) patients, respectively. Most FN (83%) and G4 neutropenia (75%) occurred in the first or second cycles. Multivariate logistic regression analyses showed that total bilirubin (TB) > the upper limit of normal range (ULN) and no dose modification from the original regimen were significantly associated with FN, and that TB > ULN, no dose modification from the original regimen, low platelet count (<15×104/μl), and recurrent disease after pancreatectomy were independent risk factors for G4 neutropenia. No dose modification from the original regimen and TB > ULN were risk factors for FN and G4 neutropenia.


Characterization of vestibular dysfunction in the mouse model for Usher syndrome 1F.
The deaf-circling Ames waltzer (av) mouse harbors a mutation in the protocadherin 15 (Pcdh15) gene and is a model for inner ear defects associated with Usher syndrome type 1F. Earlier studies showed altered cochlear hair cell morphology in young av mice. In contrast, no structural abnormality consistent with significant vestibular dysfunction in young av mice was observed. Light and scanning electron microscopic studies showed that vestibular hair cells from presumptive null alleles Pcdh15(av-Tg) and Pcdh15(av-3J) are morphologically similar to vestibular sensory cells from control littermates, suggesting that the observed phenotype in these alleles might be a result of a central, rather than peripheral, defect. In the present study, a combination of physiologic and anatomic methods was used to more thoroughly investigate the source of vestibular dysfunction in Ames waltzer mice. Analysis of vestibular evoked potentials and angular vestibulo-ocular reflexes revealed a lack of physiologic response to linear and angular acceleratory stimuli in Pcdh15 mutant mice. Optokinetic reflex function was diminished but still present in the mutant animals, suggesting that the defect is primarily peripheral in nature. These findings indicate that the mutation in Pcdh15 results in either a functional abnormality in the vestibular receptor organs or that the defects are limited to the vestibular nerve. AM1-43 dye uptake has been shown to correlate with normal transduction function in hair cells. Dye uptake was found to be dramatically reduced in Pcdh15 mutants compared to control littermates, suggesting that the mutation affects hair cell function, although structural abnormalities consistent with significant vestibular dysfunction are not apparent by light and scanning electron microscopy in the vestibular neuroepithelia of young animals.


Role of Ia antigen expression and secretory function of accessory cells in the induction of cytotoxic T lymphocyte responses against herpes simplex virus.
Splenocytes from mice which have been primed in vivo with herpes simplex virus type 1 (HSV-1) can be restimulated in vitro with infectious or UV-inactivated HSV-1 to generate HSV-specific, H-2-restricted cytotoxic T lymphocytes (CTL). HSV-primed splenocytes which have been depleted of adherent cells by sequential incubation on plastic, nylon wool, and Sephadex G-10 are not able to respond with a CTL response when restimulated in vitro. A variety of Ia-positive and Ia-negative (Ia(+) and Ia(-)) cell populations were assessed for their ability to supply accessory cell functions to spleen cells which had been exhaustively depleted of adherent cells, as measured by the restoration of a CTL response to HSV-1. Of these, only those populations which included Ia(+) cells were capable of providing accessory cell function. The ineffective populations were devoid of Ia antigens, except for B lymphocytes, which are Ia(+) and still incapable of serving as accessory cells. Splenic adherent cells and resident peritoneal cells were both proficient at restoring anti-HSV CTL responses, although splenic adherent cells were clearly superior at limiting cell numbers. Neither population was capable of accessory cell activity, however, if it was pretreated with anti-Ia antiserum plus complement or if anti-Ia serum was present during induction. Peritoneal cells lose virtually all of their membrane-associated Ia antigen after a brief period of in vitro culture (24 to 48 h). Cultured peritoneal cells, as well as P388D(1) cells (normally Ia(-)), can be induced to express Ia antigens within 48 h if they are cultured with concanavalin A-stimulated spleen cell supernatants. Ia(+) P388D(1) cells without the extraneous macrophage factor are not able to restore CTL responsiveness to HSV-1 in vitro, whereas Ia(+) cultured macrophages are fully competent accessory cells. When Ia(+) P388D(1) cells were supplemented with the macrophage-derived soluble factor interleukin 1, they displayed a modest, but significant, capacity to restore secondary anti-HSV CTL responses. In addition, glutaraldehyde-fixed, HSV-1-infected Ia(+) peritoneal cells, which could not restore the CTL response alone, were capable of providing accessory cell function if extraneous interleukin 1 was provided. In contrast, Ia(-) cultured peritoneal cells, Ia(-) P388D(1) cells, and various other Ia(-) cell lines were unable to participate in the generation of CTL even in the presence of interleukin 1. The adherent cell population would therefore appear to be making at least two essential contributions to the process of CTL development, namely, the secretion of interleukin 1 and the presentation of antigen in the context of membrane-associated Ia antigen.


CBX4 promotes antitumor immunity by suppressing Pdcd1 expression in T cells.
E3 SUMO-protein ligase CBX4 (CBX4), a key component of polycomb-repressive complexes 1 (PRC1), has been reported to regulate a variety of genes implicated in tumor growth, metastasis, and angiogenesis. However, its role in T-cell-mediated antitumor immunity remains elusive. To shed light on this issue, we generated mice with T-cell-specific deletion of Cbx4. Tumor growth was increased in the knockout mice. Additionally, their tumor-infiltrating lymphocytes exhibited impaired tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) production, with an elevated programmed cell death protein 1 (PD-1) level. In fact, dysregulated Pdcd1 expression was observed in all major subsets of peripheral T cells from the knockout mice, which was accompanied by a functional defect in response to T-cell receptor (TCR) stimulation. In support of a direct link between CBX4 and PD-1, Cbx4 overexpression resulted in the downregulation of Pdcd1 expression. Epigenetic analyses indicated that Cbx4 deficiency leads to diminished accumulation of inhibitory histone modifications at conserved region (CR)-C and CR-B sites of the Pdcd1 promoter, namely mono-ubiquitinated histone H2A at lysine 119 (H2AK119ub1) and trimethylated histone H3 at lysine 27 (H3K27me3). Moreover, inhibition of either the E3 ligase activity of polycomb-repressive complexes 1 (PRC1) or the methyltransferase activity of polycomb-repressive complexes 2 (PRC2) restores Pdcd1 expression in Cbx4-transfected cells. Cumulatively, this study reveals a novel function of CBX4 in the regulation of T-cell function and expands our understanding of the epigenetic control of Pdcd1 expression.


Coronary revascularization in lung transplant recipients with concomitant coronary artery disease.
Coronary artery disease (CAD) is not uncommon among lung transplant candidates. Several small, single-center series have suggested that short-term outcomes are acceptable in selected patients who undergo coronary revascularization prior to, or concomitant with, lung transplantation. Our objective was to evaluate perioperative and intermediate-term outcomes in this patient population at our institution. We performed a retrospective, observational cohort analysis of 898 lung transplant recipients between 1997 and 2010. Pediatric, multivisceral, lobar or repeat transplantations were excluded, resulting in 791 patients for comparative analysis, of which 49 (median age 62, 79.6% bilateral transplant) underwent concurrent coronary artery bypass and 38 (median age 64, 63.2% bilateral transplant) received preoperative percutaneous coronary intervention (PCI). Perioperative mortality, overall unadjusted survival and adjusted hazard ratio for cumulative risk of death were similar among both revascularization groups as well as controls. The rate of postoperative major adverse cardiac events was also similar among groups; however, concurrent coronary artery bypass was associated with longer postoperative length of stay, more time in the intensive care unit and more postoperative days requiring ventilator support. These results suggest that patients with CAD need not be excluded from lung transplantation. Preferential consideration should be given to preoperative PCI when feasible.


Influence of the association between simvastatin and demineralized bovine bone matrix on bone repair in rats.
Simvastatin, a drug used to lower blood cholesterol, has been reported to have an anabolic effect on bone. The purpose of this study was to evaluate the influence of simvastatin and demineralized bovine bone matrix (DBBM) on the repair of rat calvarial defects. Defects of 5 mm were created in 64 rats, divided into four groups: no local treatment (control); treatment with DBBM (DBBM); treatment with a combination of simvastatin solution (2.2 mg/50 μl) and DBBM (DBBMSIM-1); and treatment with simvastatin solution (0.5 mg/50 μl) and DBBM (DBBMSIM-2). Animals were sacrificed on postoperative day 30 or 60, after which the calvariae were X-rayed and prepared for histomorphometric evaluation. The data were submitted to statistical analysis (p < 0.05). X-rays revealed that, on postoperative day 30, animals treated with a lower dose of simvastatin presented the lowest bone density, whereas on postoperative day 60 the use of simvastatin, regardless of the dose, resulted in lower density than that observed in control and DBBM group samples. Histomorphometric analysis revealed that, on postoperative day 30, both DBBM and DBBMSIM-1 had a negative impact on bone formation. On postoperative day 60, none of the combinations tested impaired bone repair. These results showed that the association between DBBM and simvastatin had a negative impact on bone repair.


Quantifying trace element and isotope fluxes at the ocean-sediment boundary: a review.
Quantifying fluxes of trace elements and their isotopes (TEIs) at the ocean's sediment-water boundary is a pre-eminent challenge to understand their role in the present, past and future ocean. There are multiple processes that drive the uptake and release of TEIs, and properties that determine their rates are unevenly distributed (e.g. sediment composition, redox conditions and (bio)physical dynamics). These factors complicate our efforts to find, measure and extrapolate TEI fluxes across ocean basins. GEOTRACES observations are unveiling the oceanic distributions of many TEIs for the first time. These data evidence the influence of the sediment-water boundary on many TEI cycles, and underline the fact that our knowledge of the source-sink fluxes that sustain oceanic distributions is largely missing. Present flux measurements provide low spatial coverage and only part of the empirical basis needed to predict TEI flux variations. Many of the advances and present challenges facing TEI flux measurements are linked to process studies that collect sediment cores, pore waters, sinking material or seawater in close contact with sediments. However, such sampling has not routinely been viable on GEOTRACES expeditions. In this article, we recommend approaches to address these issues: firstly, with an interrogation of emergent data using isotopic mass-balance and inverse modelling techniques; and secondly, by innovating pursuits of direct TEI flux measurements. We exemplify the value of GEOTRACES data with a new inverse model estimate of benthic Al flux in the North Atlantic Ocean. Furthermore, we review viable flux measurement techniques tailored to the sediment-water boundary. We propose that such activities are aimed at regions that intersect the GEOTRACES Science Plan on the basis of seven criteria that may influence TEI fluxes: sediment provenance, composition, organic carbon supply, redox conditions, sedimentation rate, bathymetry and the benthic nepheloid inventory.This article is part of the themed issue 'Biological and climatic impacts of ocean trace element chemistry'.


Exploring the role of the metabolite-sensing receptor GPR109a in diabetic nephropathy.
Alterations in gut homeostasis may contribute to the progression of diabetic nephropathy. There has been recent attention on the renoprotective effects of metabolite-sensing receptors in chronic renal injury, including the G protein-coupled receptor (GPR)109a, which ligates the short-chain fatty acid butyrate. However, the role of GPR109a in the development of diabetic nephropathy, a milieu of diminished microbiome-derived metabolites, has not yet been determined. The present study aimed to assess the effects of insufficient GPR109a signaling, via genetic deletion of GPR109a, on the development of renal injury in diabetic nephropathy. Gpr109a-/- mice or their wild-type littermates (Gpr109a+/+) were rendered diabetic with streptozotocin. Mice received a control diet or an isocaloric high-fiber diet (12.5% resistant starch) for 24 wk, and gastrointestinal permeability and renal injury were determined. Diabetes was associated with increased albuminuria, glomerulosclerosis, and inflammation. In comparison, Gpr109a-/- mice with diabetes did not show an altered renal phenotype. Resistant starch supplementation did not afford protection from renal injury in diabetic nephropathy. While diabetes was associated with alterations in intestinal morphology, intestinal permeability assessed in vivo using the FITC-dextran test was unaltered. GPR109a deletion did not worsen gastrointestinal permeability. Furthermore, 12.5% resistant starch supplementation, at physiological concentrations, had no effect on intestinal permeability or morphology. The results of this study indicate that GPR109a does not play a critical role in intestinal homeostasis in a model of type 1 diabetes or in the development of diabetic nephropathy.


Gender and racial favouritism in black and white preschool girls.
The authors examined gender and racial preferential behaviour in 108 3- and 5-year-old Black and White girls. Children set up a birthday party for dolls that differed in gender and racial physical characteristics. Whereas White girls showed favouritism towards the doll most closely resembling themselves in both gender and race, Black girls showed most favouritism towards the White girl doll. Black girls were more likely to show preference based on gender rather than race, whereas White girls were equally likely to show race- or gender-based favouritism. Among White 5-year-olds, greater prior interaction with Blacks was positively associated with race-related favouritism (i.e., secondary preference to the White boy doll rather than the Black girl doll). Interracial contact was unrelated to racial favouritism among the other three groups. Results demonstrate the salience of gender identity during the preschool years, and indicate that majority/minority status and intergroup contact shape the development of collective identity and social behaviour.


Bcr/abl+ autologous dendritic cells for vaccination in chronic myeloid leukemia.
In chronic myeloid leukemia (CML) ex vivo generated DC are characterized by constitutive expression of bcr/abl and possibly other yet undefined leukemia-associated antigens, since these DC share a common progeny with leukemic cells. Induction of anti-leukemic T cell responses has been described in vitro. For a phase I vaccination study, autologous bcr/abl+ DC are generated under GMP conditions mainly from monocyte precursors in chronic phase CML patients. Lin-, CD80+, CD86+, CD83+, DR+ DC could be generated in sufficient numbers for s.c. vaccination with 1 x 10(6)-5 x 10(7) DC. Using monocyte precursors, the yield of DC per seeded PBMC was in the range of 1-6%. Furthermore, we could demonstrate in vitro that the T cell stimulatory ability of CD34+-derived DC can be augmented by a factor 2-3 by retroviral transduction with a gene coding for interleukin-7. DC-based vaccination strategies are a promising clinical approach, particularly as postremission immunotherapy in the setting of autologous stem cell transplantation.


Novel cluster of alpha-interferon gene sequences in a placental cosmid DNA library.
A human cosmid library was constructed and probed with a human alpha interferon (IFN-alpha) cDNA clone. One clone giving a strong hybridizing signal was isolated and characterized. The cosmid DNA insert represents a section of the human genome containing three regions of IFN-alpha-like sequences. The DNA was characterized with restriction endonuclease mapping, thereby allowing comparison to similar linkage groups reported recently and determination of homologous regions on the known physical map. The three IFN-alpha-like sequences were analyzed by a partial sequence analysis. Mapping and sequence data establish this section as a not-yet-described cluster of IFN-alpha sequences in the human genome; however, a part of the section matches to some degree to a previously described genomic region. The region described here could represent genetic polymorphism or a duplicated segment.


Acquired aquagenic papulotranslucent acrokeratoderma: report of two cases.
Aquagenic papulotranslucent acrokeratoderma (APA) is an acquired, unusual condition characterized by bilateral, symmetric, translucent-to-white papules located on the hands and feet. These lesions are accentuated following a short period of water exposure. Since its first description in 1973 as hereditary papulotranslucent acrokeratoderma, only a few cases of APA have been reported. We report two cases of teenage girls with APA. Both patients responded well to 5% salicylic acid ointment but recurrence of lesions still occurred. The close temporal proximity of presentation of these two cases may indicate that APA is not as rare as previously thought.


PACAP-(1-38) as neurotransmitter in pig pancreas: receptor activation revealed by the antagonist PACAP-(6-38).
The pituitary adenylyl cyclase-activating polypeptide PACAP-(1-38) has potent pancreatic secretory effects. We studied its immunohistochemical localization, release, and contribution to secretion induced by electrical vagus stimulation using isolated perfused porcine pancreas and the PACAP receptor antagonist PACAP-(6-38) (10(-7) M). PACAP was found in nerve fibers throughout the pancreas but, in particular, encircling ganglionic vasoactive intestinal polypeptide (VIP)-positive nerve cell bodies and, mostly, colocalized with VIP. Vagus stimulation caused its release. PACAP-(1-38)(4 x 10(-9) M) stimulated exocrine and endocrine secretion and released VIP. PACAP-(6-38) decreased PACAP-induced flow of juice to 59 +/- 7.8% and insulin secretion and VIP release to 12 +/- 6.8 and 57 +/- 13%, respectively. Glucagon secretion was unaffected. PACAP-(6-38) reduced vagus-stimulated flow rate to 63 +/- 7.6%, insulin and glucagon responses to 31.8 +/- 13 and 6 +/- 4%, respectively, and VIP release to 23 +/- 8.4% and reduced VIP-induced (2 x 10(-9) M) juice and insulin (but not glucagon) outputs to 8.3 +/- 4.2 and 67 +/- 14%, respectively. In conclusion, 1) pancreatic PACAP fibers seem to activate intrapancreatic VIPergic neurons, 2) PACAP-(6-38) antagonism documents the role of VIP/PACAP for neural regulation but cannot distinguish their relative importance, and 3) a PACAP receptor with low affinity for PACAP-(6-38), associated with glucagon cells, may exist.


Impact of structurally diverse polysaccharides on colonic mucin O-glycosylation and gut microbiota.
Understanding how dietary polysaccharides affect mucin O-glycosylation and gut microbiota could provide various nutrition-based treatments. Here, the O-glycan profile of the colonic mucosa and gut microbiome were investigated in C57BL/6J mice fed six structurally diverse dietary polysaccharides and a mixture of six fibers. Dietary polysaccharides increased total O-glycans, mainly by stimulating neutral glycans. Highly branched arabinogalactan promoted terminally fucosylated core 1 O-glycans; whereas linear polysaccharides, including pectin, konjac glucomannan, inulin, and the fiber mixture, favored terminally di-fucosylated O-glycans. The last three polysaccharides also lowered the level of sulfated O-glycans and sialylated mono-fucosylated O-glycans. Varied monosaccharide composition in mixed polysaccharides had a synergistic beneficial effect, boosting fucosylated neutral glycans, decreasing acidic glycans, and stimulating microbial richness and diversity. Dietary polysaccharides containing arabinose and sulfate groups enhanced the relative abundances of Akkermansia and Muribaculaceae, respectively. The present comparison reveals the relationship between dietary polysaccharide structure, mucin O-glycan composition, and intestinal microorganisms.


Critical role of truncated α-synuclein and aggregates in Parkinson's disease and incidental Lewy body disease.
The role of Lewy bodies, Lewy neurites and α-synuclein (αSYN) in the pathophysiology and diagnosis of Parkinson's disease (PD) is unclear. We used postmortem human tissue, a panel of antibodies (Abs) and confocal microscopy to examine the three-dimensional neurochemical anatomy of the nigrostriatal system. Abs were specific to truncated (tαSYN), phosphorylated and full-length αSYN. The findings demonstrate the critical role of tαSYN in initiating aggregation, a role for other forms of αSYN in aggregate expansion, a reason for the wide variety of proteins present in different aggregates, an explanation for the laminar appearance of aggregates described historically using different methods, the existence of proximal greater than distal aggregation in the vulnerable nigrostriatal pathway, the independent transport of different forms of αSYN as cargo along axons and a possible sequence for the formation of Lewy bodies. Findings differed between incidental Lewy body disease and PD only quantitatively. These findings have implications for understanding the pathogenesis and treatment of PD.


Primary structures of three human neutrophil defensins.
The primary structures of three human neutrophil antimicrobial peptides (HNP) were determined. The peptides, HNP-1, HNP-2, and HNP-3, which we have termed defensins, were rich in cystine, arginine, and aromatic residues, but were devoid of free sulfhydryl groups and carbohydrate moieties. They were 29-30 residues in length and identical in sequence in all but their amino terminal residues. The defensins were homologous in sequence to peptides of similar size and biological activity previously purified from rabbit polymorphonuclear leukocytes, but unrelated to other neutrophil proteins of known sequence. 11 amino acid residues of the human defensins, including all six cysteinyl residues, were invariantly conserved in the six rabbit members of this multigene peptide family. That similarly structured antimicrobial peptides are present in both rabbit and human leukocytes supports their purported role as cidal agents in phagocyte-mediated host defense.


p53 mutations in leukemia and myelodysplastic syndrome after ovarian cancer.
Although p53 mutations occur in alkylating agent-related leukemias, their frequency and spectrum in leukemias after ovarian cancer have not been addressed. The purpose of this study was to examine p53 mutations in leukemias after ovarian cancer, for which treatment with platinum analogues was widely used. Adequate leukemic or dysplastic cells were available in 17 of 82 cases of leukemia or myelodysplastic syndrome that occurred in a multicenter, population-based cohort of 23,170 women with ovarian cancer. Eleven of the 17 received platinum compounds and other alkylating agents with or without DNA topoisomerase II inhibitors and/or radiation. Six received other alkylating agents, in one case, with radiation. Genomic DNA was extracted and p53 exons 5, 6, 7, and 8 were amplified by PCR. Mutations and loss of heterozygosity were analyzed on the WAVE instrument (Transgenomic) followed by selected analysis by sequencing. Eleven p53 mutations involving all four exons studied and one polymorphism were identified. Genomic DNA analyses were consistent with loss of heterozygosity for four of the mutations. The 11 mutations occurred in 9 cases, such that 6 of 11 leukemias after platinum-based regimens (55%) and 3 of 6 leukemias after other treatments (50%) contained p53 mutations. Two leukemias that occurred after treatment with platinum analogues contained two mutations. Among eight mutations in leukemias after treatment with platinum analogues, there were four G-to-A transitions and one G-to-C transversion. p53 mutations are common in leukemia and myelodysplastic syndrome after multiagent therapy for ovarian cancer. The propensity for G-to-A transitions may reflect specific DNA damage in leukemias after treatment with platinum analogues.


Use of a highly sensitive assay to analyze the excision repair of dimer and nondimer DNA damages induced in human skin fibroblasts by 254-nm and solar ultraviolet radiation.
The excision repair of nondimer DNA damages induced in normal human skin fibroblasts exposed to the Mylar-filtered UV produced by a fluorescent sunlamp was investigated. This work was accomplished through the development of a modification of the bromodeoxyuridine photolysis assay that greatly increases the sensitivity of this assay. This enhancement in sensitivity was achieved through use of alkaline elution to measure the DNA strand breakage produced by the photolysis of bromodeoxyuridine incorporated into the DNA through excision repair. Using this modified bromodeoxyuridine photolysis assay, it was found that the solar UV-induced nondimer DNA damages appear to have been repaired by a short patch repair mechanism in which a small number of nucleotides (two to four) were inserted into the repaired site. This is in contrast to the long patch repair process involved in the excision of cyclobutane pyrimidine dimers in which approximately 40 nucleotides were inserted into each repaired region.


High-fidelity digital recording and playback sphygmomanometry system: device description and proof of concept.
This study describes the development of a new digital sphygmocorder (DS-II), which allows the digital recording and playback of the Korotkoff sounds, together with cuff pressure waveform, and its performance in a pilot validation study. A condenser microphone and stethoscope head detect Korotkoff sounds and an electronic chip, dedicated to audio-signal processing, is used to record high-quality sounds. Systolic and diastolic blood pressure (SBP/DBP) are determined from the recorded signals with an automatic beat detection algorithm that displays the cuff pressure at each beat on the monitor. Recordings of Korotkoff sounds, with the cuff pressure waveforms, and the simultaneous on-site assessments of SBP/DBP were performed during 100 measurements in 10 individuals. The observers reassessed the recorded signals to verify their accuracy and differences were calculated. The features of the high-fidelity DS-II, the technical specifications and the assessment procedures utilizing the playback software are described. Interobserver absolute differences (mean±SD) in measurements were 0.7±1.1/1.3±1.3 mmHg (SBP/DBP) with a mercury sphygmomanometer and 0.3±0.9/0.8±1.2 mmHg with the DS-II. The absolute DS-II mercury sphygmomanometer differences were 1.3±1.9/1.5±1.3 mmHg (SBP/DBP). The high-fidelity DS-II device presents satisfactory agreement with simultaneous measurements of blood pressure with a mercury sphygmomanometer. The device will be a valuable methodology for validating new blood pressure measurement technologies and devices.


Nickel: A micronutrient element for hydrogen-dependent growth of Rhizobium japonicum and for expression of urease activity in soybean leaves.
Soybean plants and Rhizobium japonicum 122 DES, a hydrogen uptake-positive strain, were cultured in media purified to remove Ni. Supplemental Ni had no significant effect on the dry matter or total N content of plants. However, the addition of Ni to both nitrate-grown and symbiotically grown plants resulted in a 7- to 10-fold increase in urease activity (urea amidohydrolase, EC 22.214.171.124) in leaves and significantly increased the hydrogenase activity (EC 188.8.131.52) in isolated nodule bacteroids. When cultured under chemolithotrophic conditions, free-living R. japonicum required Ni for growth and for the expression of hydrogenase activity. Hydrogenase activity was minimal or not detectable in cells incubated either without Ni or with Ni and chloramphenicol. Ni is required for derepression of hydrogenase activity and apparently protein synthesis is necessary for the participation of Ni in hydrogenase expression. The addition of Cr, V, Sn, and Pb in place of Ni failed to stimulate the activity of hydrogenase in R. japonicum and urease in soybean leaves. The evidence indicates that Ni is an important micronutrient element in the biology of the soybean plant and R. japonicum.


Self-assembly and lipid interactions of diacylglycerol lactone derivatives studied at the air/water interface.
Synthetic diacylglycerol lactones (DAG-lactones) have been shown to be effective modulators of critical cellular signaling pathways. The biological activity of these amphiphilic molecules depends in part upon their lipid interactions within the cellular plasma membrane. This study explores the thermodynamic and structural features of DAG-lactone derivatives and their lipid interactions at the air/water interface. Surface-pressure/area isotherms and Brewster angle microscopy revealed the significance of specific side-groups attached to the terminus of a very rigid 4-(2-phenylethynyl)benzoyl chain of the DAG-lactones, which affected both the self-assembly of the molecules and their interactions with phospholipids. The experimental data highlight the formation of different phases within mixed DAG-lactone/phospholipid monolayers and underscore the relationship between the two components in binary mixtures of different mole ratios. Importantly, the results suggest that DAG-lactones are predominantly incorporated within fluid phospholipid phases rather than in the condensed phases that form, for example, by cholesterol. Moreover, the size and charge of the phospholipid headgroups do not seem to affect DAG-lactone interactions with lipids.


Common peroneal nerve palsy after robotic assisted laparoscopic colectomy: A case report.
Common peroneal nerve palsy after surgery with lithotomy position has been widely reported, but it is an unexpected complication after surgery with supine position. We report a patient who developed common peroneal nerve palsy after surgery with supine position. A 55-year old man is planed for robotic assisted laparoscopic right hemicolectomy because of colon cancer. The patient was placed supine with Trendelenburg position at an angle about 5 degrees and tilted left about 15 degrees. The operation is uneventful, but he developed common peroneal nerve palsy on the first postoperative day. The patient was fully recovered with conservative treatment after 2 months. We consider that nerve palsy as a result of compression of common peroneal nerve related to patient positioning. So we should be careful not to develop common peroneal nerve palsy even if the patient was placed in the supine position during robotic assisted surgery.


[Labor relations under decentralized health management and dilemmas in the relationship between work expansion and casualization in the Brazilian Unified National Health System].
This article analyzes the logic behind human resources management in primary health care from the perspective of municipal health systems administrators in Brazil, rethinking the decentralization process based on mediation between Federal government guidelines and the health services' daily practice. This was a cross-sectional study using a semi-structured questionnaire applied to municipal health secretaries in the Southeastern Health Macro-Region (Minas Gerais State). The municipal health secretaries had a mean age of 39.4 years and low schooling, and a large proportion did not belong to the health field. As for the workforce in the Family Health Program, the municipal governments hired 93.3% of the health professionals, mostly on temporary contracts. The findings confirm gaps in the workers' political and social rights. Paradoxically, the government itself, through the municipal administration, is responsible for failing to comply with the prevailing labor legislation.


Bacillus Calmette-Guerin (BCG) therapy is safe and effective in non-muscle invasive bladder cancer (NMIBC) patients with immunomodulating conditions.
Bacillus Calmette-Guerin (BCG) is the most effective therapy available to treat high-risk nonmuscle invasive bladder cancer (NMIBC) patients. However, for patients with immunomodulating conditions BCG is a relative contraindication due to efficacy and safety concerns. To our knowledge, no population-level study evaluating the efficacy and safety profile of BCG for immunomodulated patients exists. NMIBC patients aged 66 years or older were identified in the Surveillance, Epidemiology, and End Results (SEER) - Medicare database from 1975-2013. All patients completed adequate BCG (at least 5 plus 2 treatments completed within 12 months of diagnosis). Two groups were defined: an immunomodulated population identified by immunomodulating conditions such as solid-organ transplantation, HIV, and autoimmune conditions, and an immunocompetent group. The primary endpoint was 5-year progression-free survival defined as progression to systemic chemotherapy, checkpoint inhibitors, radical or partial cystectomy, metastasis, or cancer-specific death. A safety analysis was performed as a secondary outcome. In a total of 4,277 patients with NMIBC who completed adequate BCG, 606 (14.2%) were immunomodulated. The immunomodulated group was older at diagnosis (P < 0.001), more likely to be female (P < 0.001), more likely to live in a metropolitan area (P < 0.001), and had higher Charlson comorbidity scores (P < 0.001). There were no differences in progression to chemotherapy (P = 0.17), checkpoint inhibitors (P > 0.99), radical cystectomy (P = 0.40), partial cystectomy (P = 0.93), metastasis (P = 0.19), cancer-specific death (P = 0.18) or 5-year total bladder cancer progression (P = 0.30) between the groups. For the safety analysis, rates of disseminated BCG were similar between immunomodulated and immunocompetent patients (0.7% vs. <1.8%, P = 0.51). On multivariable analysis 5-year total bladder cancer progression (HR 1.07 [CI 0.88-1.30]) was similar between the groups. Rates of bladder cancer progression and disseminated BCG complications 5-years after BCG therapy were similar regardless of immunomodulation status. These findings suggest that BCG intravesical therapy can be offered to immunomodulated patients with high-risk NMIBC although theoretical infectious complication risks remain.


A calcium wave mediated by gap junctions coordinates a rhythmic behavior in C. elegans.
Intercellular calcium waves can be observed in adult tissues, but whether they are instructive, permissive, or even required for behavior is predominantly unknown. In the nematode Caenorhabditis elegans, a periodic calcium spike in a pacemaker cell initiates a calcium wave in the intestine. The calcium wave is followed by three muscle contractions that comprise the defecation motor program. Normal wave propagation requires the pannexin gap-junction subunit INX-16 at the interfaces of the intestinal cells. In the absence of this gap-junction subunit, calcium waves are frequently absent. The remaining waves are slow, initiate at abnormal locations, or travel in the opposite direction. Abnormal waves are associated with parallel effects in the first step of the motor program: The contractions of the overlying muscles fail to propagate beyond the pacemaker cell, are slow, initiate in abnormal locations, or are reversed. Moreover, the last two motor steps are predominantly absent. Finally, the absence of this gap-junction subunit also affects the reliability of the pacemaker cell; cycle timing is often irregular. These data demonstrate that pannexin gap junctions propagate calcium waves in the C. elegans intestine. The calcium waves instruct the motor steps and regulate the pacemaker cell's authority and reliability.


Proportions of circulating T cells with a regulatory cell phenotype increase with HIV-associated immune activation and remain high on antiretroviral therapy.
To examine the relationships between blood CD4 natural regulatory T (Treg) cells, plasma HIV RNA level, CD4 T-cell count and immune activation in untreated HIV-infected patients and immunodeficient patients beginning antiretroviral therapy (ART), using a novel phenotype to define Treg cells (CD25CD127CD4). Data were compared with established Treg cell markers (FoxP3, CTLA-4 and GITR). Twenty-nine untreated HIV-infected patients with CD4 T-cell counts of < 300 or > 400/microl were compared in a cross-sectional study and 12 patients beginning combination ART with < 100 CD4 T cells/mul were followed for 1 year on therapy. Three- and four-colour flow cytometry was used to quantitate proportions of Treg cells. In control donors and patients with high CD4 T-cell counts, 28-89% (median 60%) of CD25CD127CD4 cells were FoxP3, but < 10% expressed GITR or CTLA-4. Immunodeficient patients also had CD4-negative lymphocytes with the phenotype FoxP3CD127. Proportions of CD25CD127 cells and activated (HLA-DR) cells in the CD4 T-cell population were increased in patients with low CD4 T cell counts. The proportion of CD25CD127CD4 T cells correlated positively with plasma HIV RNA level and CD4 T-cell activation, but inversely with CD4 T-cell count. Longitudinal studies of 12 patients receiving ART in two distinct cohorts (Western Australia and Malaysia) showed that the proportion of CD25CD127CD4 cells decreased slightly over time, but remained above levels seen in non-HIV controls. Proportions of circulating T cells with a regulatory cell phenotype increase with HIV-associated immune activation and remain high after 1 year on ART.


A Redox-Inactive Derivative of Tocotrienol Suppresses Tumor Growth of Mesothelioma Cells in a Xenograft Model.
Malignant mesothelioma (MM) is an aggressive cancer with poor prognosis. We focused on the anticancer activity of tocotrienol (T3) and have reported that a new redox-inactive T3 derivative (6-O-carboxypropyl-α-tocotrienol; T3E) exerts stronger inhibitory effects on MM cell growth than that of T3 in vitro. Furthermore, we have revealed some mechanisms of T3E that are involved in anti-MM effects. However, the effect of T3E in vivo remains unclear. In this study, we compared the plasma concentrations of T3E to that of T3 using mice to clarify differences in pharmacokinetics. Blood was sequentially collected after oral administration of T3 or T3E, and plasma concentrations were analyzed by HPLC. The area under the plasma T3 and T3E concentration-time curve from 0 to 24 h (AUC0-24 h) of T3E was two times higher than that of T3. In addition, we evaluated the effect of T3E oral administration on tumor growth using a xenograft model of mice that were transplanted with human MM cells (H2052 cell line). Tumor volume was significantly reduced without body weight loss in mice orally administered 150 mg/kg T3E once per 2 d for 10 d, which suggests that T3E has potential anti-MM effects.


Correlation between clinical outcome and size and site of the lesion in computed tomography guided thalamotomy and pallidotomy.
Fourteen thalamotomies and five pallidotomies were performed in 19 patients with hereditary intention tremor or Parkinson's disease. The target coordinates were determined by a stereotactic computed tomography study using the Laitinen noninvasive stereoadapter. Surgery was done without ventriculography. The patients were assessed 3-12 months later. In a postoperative stereotactic computed tomography study, the positions of the thalamic and pallidal targets were marked, and the coordinates of the center of the lesion were measured in relation to these targets. The volume of the lesion was calculated. In 3 thalamic lesion patients, no lesion could be visualized. The size of the eleven visible thalamic lesions ranged from 4 to 75 mm3 (mean 26), and the size of the 5 pallidal lesions ranged from 28 to 150 mm3 (mean 67). On the average, the center of the lesion was 1.4 mm medial to the position of the anatomical target (p less than 0.002). Neither size nor site of the lesion correlated with the clinical outcome.


A limited unilateral transpedicular approach for anterior decompression of the thoracolumbar spinal cord in elderly and high-risk patients.
Surgical treatment for elderly patients with thoracolumbar (TL) kyphosis and spinal cord (SC) compression presents significant challenges due to compression location, the amount of deformity, and patient's medical status might not permit full correction of the deformity. In this series, we present a surgical approach that provides adequate decompression without the risks associated with a pedicle subtraction osteotomy/posterior vertebral column resection or an anterior corpectomy. Three patients presented with TL kyphosis and progressive neurologic symptoms. All had acute weakness; none were ambulatory. SC was compressed over the apex of kyphosis, and for some, there was spinal stenosis at the proximal junction of the TL spine. The surgical technique involved unilateral resection of the pars, pedicles, the posterior one-third of the lateral wall of the vertebral body, decancellation of the impinging kyphus, and finally resection of the posterior vertebral body wall compressing the SC followed by instrumentation and fusion two levels above and below the fused segments. All patients survived the procedure and left the hospital after 10-22 days. Estimated blood loss was 653 ml. No deep infections occurred. One patient developed acute tubular necrosis but recovered fully. The other two showed improvement of one Frankel grade and were independent in the final follow-up. One patient developed acute tubular necrosis but recovered fully yet his neurologic status was unchaged. The other two showed improvement of one Frankel grade and were independent in the final follow-up. The procedure described presents a compromise that fits the more elderly patient that might not be able to tolerate major deformity correction and at the same time provides similar results in the short and medium term to more extensive procedures.