Tundra wildfire triggers sustained lateral nutrient loss in Alaskan Arctic.
Climate change is creating widespread ecosystem disturbance across the permafrost zone, including a rapid increase in the extent and severity of tundra wildfire. The expansion of this previously rare disturbance has unknown consequences for lateral nutrient flux from terrestrial to aquatic environments. Lateral loss of nutrients could reduce carbon uptake and slow recovery of already nutrient-limited tundra ecosystems. To investigate the effects of tundra wildfire on lateral nutrient export, we analyzed water chemistry in and around the 10-year-old Anaktuvuk River fire scar in northern Alaska. We collected water samples from 21 burned and 21 unburned watersheds during snowmelt, at peak growing season, and after plant senescence in 2017 and 2018. After a decade of ecosystem recovery, aboveground biomass had recovered in burned watersheds, but overall carbon and nitrogen remained ~20% lower, and the active layer remained ~10% deeper. Despite lower organic matter stocks, dissolved organic nutrients were substantially elevated in burned watersheds, with higher flow-weighted concentrations of organic carbon (25% higher), organic nitrogen (59% higher), organic phosphorus (65% higher), and organic sulfur (47% higher). Geochemical proxies indicated greater interaction with mineral soils in watersheds with surface subsidence, but optical analysis and isotopes suggested that recent plant growth, not mineral soil, was the main source of organic nutrients in burned watersheds. Burned and unburned watersheds had similar δ15 N-NO3 - , indicating that exported nitrogen was of preburn origin (i.e., not recently fixed). Lateral nitrogen flux from burned watersheds was 2- to 10-fold higher than rates of background nitrogen fixation and atmospheric deposition estimated in this area. These findings indicate that wildfire in Arctic tundra can destabilize nitrogen, phosphorus, and sulfur previously stored in permafrost via plant uptake and leaching. This plant-mediated nutrient loss could exacerbate terrestrial nutrient limitation after disturbance or serve as an important nutrient release mechanism during succession.


Ion-selective electrode technology: an overview.
Diluting a sample before analysis, as is required by both flame photometry and indirect reading ISE analysers, gives less relevant analytical results than measuring an undiluted sample directly. This is because dilution masks the impact that protein and lipids exert on the true activity of an ion in a clinical sample. Measuring a sample directly without dilution enables only those ions which are not bound to other materials, e.g. other ions or proteins, to be detected. The levels of these free ions are more likely to reflect true physiological status than analysis of both free and bound ions. Measuring a sample of whole blood directly without dilution can only be performed accurately if the ISE analyser reference junction is unaffected by changes in haematocrit. Calibrant compositions should be of an ionic strength similar to that of physiological samples. Residual liquid junction potentials between calibrants and samples should be minimised by careful design of the reference electrode.


Does Assessment Format Matter? A Comparison of In-Person Versus Teletesting Scores for Youth with ADHD.
This study examined test score equivalency between traditional in-person assessment and teletesting among youth diagnosed with Attention-Deficit/Hyperactivity Disorder (ADHD). In all, 896 youth with ADHD, ages 5-21 years, were administered cognitive, academic achievement, and verbal fluency measures via either teletesting (n = 448) or traditional in-person assessment (n = 448). The teletesting and in-person groups were matched on age, sex, and insurance type (as a proxy for income). Results indicated no significant differences in test scores obtained via in-person and teletesting evaluations across all examined measures. Clinically referred youth with ADHD perform similarly on measures of cognitive functioning, academic achievement, and verbal fluency, regardless of whether these measures are administered in-person or via teletesting. While additional evidence for equivalent psychometric properties of neuropsychological instruments administered remotely is needed, this study offers support for the validity of remote administration among youth with ADHD.


Short-Term Perinatal Outcome among Term Infants with Prenatal Diagnosis of Large Abdominal Circumference.
Background Measuring fetal abdominal circumference (AC) prenatally is an effective tool for predicting neonatal weight and macrosomia. Data are lacking regarding the outcome of newborn infants with prenatal diagnosis of large AC. Aim The aim of this study was to evaluate early short-term neonatal outcome among term singleton newborn infants with prenatal diagnosis of large AC. Methods Retrospective data were collected on 501 term infants with prenatal diagnosis of large AC (≥ 360 mm) and on matched controls, including information on maternal condition and on infant perinatal complications. Results In compare with controls, the study group had higher incidence of macrosomia (188 [37.5%] vs. 18 [3.6%], p < 0.001), hypoglycemia (48 (9.6%) vs. 25 [5%], p = 0.007), and significant morbidity (49 [9.8%] vs. 28 [5.6%], p = 0.017) but without increased incidence of congenital malformations or other perinatal complications. Only among the macrosomic, study subgroup and their controls differences were recorded including hypoglycemia (17.6 vs. 4.8%, p < 0.001), need for oral glucose (11.2 vs. 2.7%, p = 0.002), significant morbidity (10.1 vs. 3.7%, p = 0.024), and hospitalization in special care unit (11.7 vs. 4.3%, p = 0.012). Conclusion Prelabor diagnosis of large AC mostly reflects the infant's high birth weight and macrosomia with the associated perinatal complications. Large AC by itself was not predictive of any congenital malformations or perinatal and postnatal complications.


Biomaterial microenvironments to support the generation of new neurons in the adult brain.
Neural stem cells (NSC) in two regions of the adult mammalian brain--the subventricular zone (SVZ) and hippocampus--continuously generate new neurons, enabled by a complex repertoire of factors that precisely regulate the activation, proliferation, differentiation, and integration of the newborn cells. A growing number of studies also report low-level neurogenesis in regions of the adult brain outside these established neurogenic niches--potentially via NSC recruitment or activation of local, quiescent NSCs--under perturbations such as ischemia, cell death, or viral gene delivery of proneural growth factors. We have explored whether implantation of engineered biomaterials can stimulate neurogenesis in normally quiescent regions of the brain. Specifically, recombinant versions of factors found within the NSC microenvironment, Sonic hedgehog, and ephrin-B2 were conjugated to long polymers, thereby creating highly bioactive, multivalent ligands that begin to emulate components of the neurogenic niche. In this engineered biomaterial microenvironment, new neuron formation was observed in normally non-neurogenic regions of the brain, the striatum, and the cortex, and combining these multivalent biomaterials with stromal cell-derived factor-1α increased neuronal commitment of newly divided cells seven- to eightfold in these regions. Additionally, the decreased hippocampal neurogenesis of geriatric rodents was partially rescued toward levels of young animals. We thus demonstrate for the first time de novo neurogenesis in both the cortex and striatum of adult rodents stimulated solely by delivery of synthetic biomaterial forms of proteins naturally found within adult neurogenic niches, offering the potential to replace neurons lost in neurodegenerative disease or injury as an alternative to cell implantation.


The complete mitochondrial genome of the red-jointed brackish-water fiddler crab Minuca minax (LeConte 1855) (Brachyura: Ocypodidae): New family gene order, and purifying selection and phylogenetic informativeness of protein coding genes.
Minuca minax is a semi-terrestrial crustacean that commonly lives in low salinity, riverine habitats along the shores of the eastern United States. This study reports, for the first time, the complete mitochondrial genome of M. minax. The AT-rich mitochondrial genome of M. minax is 15,937 bp in length and comprised of 13 protein-coding genes (PCGs), 2 ribosomal RNA genes, and 22 transfer RNA genes. A single 737 bp long intergenic space is assumed to be the D-loop. Most of the PCGs and tRNA genes are encoded in the L-strand. The gene order observed in the mitochondrial genome of M. minax is new although almost identical to that reported in confamiliar species. In all other confamiliar species to which M. minax is compared, the positions of the trnQ gene and the trnI gene are switched. KA/KS ratios calculated for all mitochondrial PCGs show values of <1, indicating that these PCGs are evolving under purifying selection. A maximum likelihood phylogenetic analysis (concatenated PCGs [n = 13], 15 species) supports the monophyly of the subfamilies Ocypodinae and Gelaminidae. Mitochondrial PCGs have enough phylogenetic information to reveal relationships supporting higher taxonomic levels within this family. The knowledge of a complete mitochondrial genome from the red-jointed brackish-water fiddler crab M. minax contributes to the better understanding of meta-population connectivity and the mechanisms involved in the adaptation of marine organisms to near-limnic conditions.


Lymphocytic-plasmacytic thyroiditis and glomerulonephritis in a boxer.
An 18-month-old boxer was presented for investigation of profound lethargy, and primary hypothyroidism was diagnosed. A strong antithyroglobulin antibody titre was also present, indicating lymphocytic (immune-mediated) thyroiditis as the cause of hypothyroidism. A concurrent protein-losing glomerulonephropathy was also detected, although the aetiology could not be determined at initial presentation. Thyroid replacement and dietary therapy were prescribed. The dog improved clinically for approximately 12 months when it was re-presented with poorly controlled hypothyroidism and renal failure. Postmortem examination confirmed the presence of lymphocytic-plasmacytic thyroiditis, in conjunction with membranoproliferative glomerulonephritis and renal failure.


Detection approaches for multidrug resistance genes of leukemia.
Leukemia is a clonal malignant hematopoietic stem cell disease. It is the sixth most lethal cancer and accounts for 4% of all cancers. The main form of treatment for leukemia is chemotherapy. While some cancer types with a higher incidence than leukemia, such as lung and gastric cancer, have shown a sharp decline in mortality rates in recent years, leukemia has not followed this trend. Drug resistance is often regarded as the main clinical obstacle to effective chemotherapy in patients diagnosed with leukemia. Many resistance mechanisms have now been identified, and multidrug resistance (MDR) is considered the most important and prevalent mechanism involved in the failure of chemotherapy in leukemia. In order to reverse MDR and improve leukemia prognosis, effective detection methods are needed to identify drug resistance genes at initial diagnosis. This article provides a comprehensive overview of published approaches for the detection of MDR in leukemia. Identification of relevant MDR genes and methods for early detection of these genes will be needed in order to treat leukemia more effectively.


Therapeutic benefit of radiotherapy in huge (≥10 cm) unresectable hepatocellular carcinoma.
Huge (≥10 cm) hepatocellular carcinomas (HCCs) show dismal prognosis and only a limited number of cases are eligible for curative resection. We studied the therapeutic benefit of radiotherapy (RT) in patients with huge unresectable HCCs. Data from 283 patients with huge HCCs and preserved liver function who underwent non-surgical treatment from July 2001 to March 2012 were retrospectively reviewed. Patients were divided into 4 groups according to the initial treatment: Group A (N= 49), transarterial chemoembolization (TACE); Group B (N = 35), TACE + RT; Group C (N = 50), hepatic arterial infusion chemotherapy; and Group D (n = 149), concurrent chemoradiotherapy (CCRT). The median follow-up period was 27.8 months (range, 12.9-121.9 months). The median overall survival (OS) was longer in Groups B (15.3 months) and D (12.8 months) than in Groups A (7.5 months) and C (8.2 months; Group B vs. A, Bonferroni corrected P [P(c)] = 0.04; Group B vs. C, P(c) = 0.02; Group D vs. A, P(c) = 0.01; Group D vs. C, Pc = 0.006). Groups B and D also showed superior progression-free survival (PFS) and intrahepatic control than Groups A and C. In multivariate analysis, tumour multiplicity, serum alpha-foetoprotein level (≥200 ng/ml) and initial treatment were independent prognostic factors for OS and PFS. Patients with huge unresectable HCCs treated with RT, either as CCRT or in combination with TACE, showed excellent intrahepatic control and prolonged survival. RT could be considered a promising treatment modality in these patients.


Scene-Aware Audio Rendering via Deep Acoustic Analysis.
We present a new method to capture the acoustic characteristics of real-world rooms using commodity devices, and use the captured characteristics to generate similar sounding sources with virtual models. Given the captured audio and an approximate geometric model of a real-world room, we present a novel learning-based method to estimate its acoustic material properties. Our approach is based on deep neural networks that estimate the reverberation time and equalization of the room from recorded audio. These estimates are used to compute material properties related to room reverberation using a novel material optimization objective. We use the estimated acoustic material characteristics for audio rendering using interactive geometric sound propagation and highlight the performance on many real-world scenarios. We also perform a user study to evaluate the perceptual similarity between the recorded sounds and our rendered audio.


A gated-time-of-flight top-hat electrostatic analyzer for low energy ion measurements.
When incorporated into a top-hat electrostatic analyzer, a gate electrode enables the separation of ions by their mass-per-charge with modest mass resolution (M/∆M ∼ 10). Gated-time-of-flight (TOF) instruments avoid the energy straggling and angular scattering effects prevalent in foil-based detection systems, providing more pristine measurements of three-dimensional distribution functions of incident ions. Gated-TOF implementations are ideal for measuring the properties of low-energy (i.e., <100 eV) thermal ions in various space environments. We present an instrument prototype capable of separating H+, He+, O+, and O2+ in Earth's ionosphere and demonstrate that in addition to providing species determination, precise operation of the gate electrode provides an electronically adjustable geometric factor that can extend a single instrument's dynamic range by several orders of magnitude.


SUBCLAVIAN STEAL SYNDROME: A Review.
The Subclavian Steal Syndrome is a condition that results from the stenosis or occlusion of the subclavian artery proximal to the origin of the vertebral artery. The blockage causes the reversal of the normal direction of blood flow in the vertebral artery which is termed the "steal", because it steals blood from the cerebral circulation. Blood is drawn from the contralateral vertebral, basilar or carotid artery regions into the low-pressure ipsilateral upper limb vessels. Although a relatively uncommon disease, it represents a condition which mimics many conditions often treated by chiropractors and osteopaths. The term subclavian steal syndrome is applied when reversed vertebral artery flow causes cerebral ischaemia with associated symptoms of vertebrobasilar hypoperfusion and/or symptoms of brainstem or arm ischaemia. This syndrome exists as an important consideration in the differential diagnosis of cerebral and brachial ischaemia.


Cell cycle regulation in the hyperthermophilic crenarchaeon Sulfolobus acidocaldarius.
The regulation and co-ordination of the cell cycle of the hyperthermophilic crenarchaeon Sulfolobus acidocaldarius was investigated with antibiotics. We provide evidence for a core regulation involving alternating rounds of chromosome replication and genome segregation. In contrast, multiple rounds of replication of the chromosome could occur in the absence of an intervening cell division event. Inhibition of the elongation stage of chromosome replication resulted in cell division arrest, indicating that pathways similar to checkpoint mechanisms in eukaryotes, and the SOS system of bacteria, also exist in archaea. Several antibiotics induced cell cycle arrest in the G2 stage. Analysis of the run-out kinetics of chromosome replication during the treatments allowed estimation of the minimal rate of replication fork movement in vivo to 250 bp s-1. An efficient method for the production of synchronized Sulfolobus populations by transient daunomycin treatment is presented, providing opportunities for studies of cell cycle-specific events. Possible targets for the antibiotics are discussed, including topoisomerases and protein glycosylation.


Arthroscopic findings in acute fractures of the ankle.
We have evaluated prospectively the arthroscopic findings in acute fractures of the ankle in 288 consecutive patients (148 men and 140 women) with a mean age of 45.6 years. According to the AO-Danis-Weber classification there were 14 type-A fractures, 198 type B and 76 type C. Lesions of the cartilage were found in 228 ankles (79.2%), more often on the talus (69.4%) than on the distal tibia (45.8%), the fibula (45.1%), or the medial malleolus (41.3%). There were more lesions in men than in women and in general they were more severe in men (p < 0.05). They also tended to be worse in patients under 30 years and in those over 60 years of age. The frequency and severity of the lesions increased from type-B to type-C fractures (p < 0.05). Within each type of fracture the lesions increased from subgroups 1 to 3 (p < 0.05). The anterior tibiofibular ligament was injured with increased frequency from type-B.1 to type-C3 fractures (p < 0.05), but it was not torn in all cases. While lateral ligamentous injuries were seen more often in type-B than in type-C fractures (p < 0.05), no difference was noted in the frequency of deltoid ligamentous lesions. Our findings show that arthroscopy is useful in identifying associated intra-articular lesions in acute fractures of the ankle.


The role of soft tissues in plate fixation of proximal phalanx fractures.
The tension band effect of plate fixation and the contribution of soft tissues to that effect was examined biomechanically in human proximal phalanges. Forty-six proximal phalanges in whole cadaver hands with all soft tissues in place (intact) and 43 proximal phalanges stripped of soft tissues (denuded) were tested. After midshaft osteotomy, each proximal phalanx was fixed internally with a dorsal minicondylar plate, a lateral minicondylar plate, a dorsal straight plate, or a lateral straight plate. Specimens were tested in three-point apex dorsal bending to clinical failure, defined as 30 degrees angulation. Ultimate moment (stability) at this angulation was similar among the four fixation methods in the specimens with all soft tissues intact. Stability also was similar among these methods in the denuded specimens. There were no significant differences in stability between minicondylar and straight plates or between dorsal and lateral plates in the specimens with soft tissues, nor were there significant differences between these groups in the denuded specimens. The stability of the four fixation methods was significantly greater in the specimens with soft tissues than in the denuded specimens. Soft tissues increased the stability of lateral minicondylar plates by 163%, lateral straight plates by 157%, dorsal minicondylar plates by 126%, and dorsal straight plates by 104%, providing a dorsal tension band effect that counteracted the buttress (compression) of the volar fracture surfaces of the phalanx. The results suggest that in the clinical setting a laterally placed straight or minicondylar plate may provide as much stability to a phalanx with a midshaft fracture as does the traditional, more invasive dorsally placed minicondylar or straight plate. These findings must be evaluated with caution, however, because all specimens were from embalmed cadavers, and the formalin fixation may have augmented the stability and stiffness of the soft tissues in the intact specimens. A subsequent pilot study comparing intact proximal phalangeal specimens that were formalin-fixed with those that were fresh-frozen showed a significant increase in stability and stiffness of formalin-fixed specimens.


An ultraslow-spreading class of ocean ridge.
New investigations of the Southwest Indian and Arctic ridges reveal an ultraslow-spreading class of ocean ridge that is characterized by intermittent volcanism and a lack of transform faults. We find that the mantle beneath such ridges is emplaced continuously to the seafloor over large regions. The differences between ultraslow- and slow-spreading ridges are as great as those between slow- and fast-spreading ridges. The ultraslow-spreading ridges usually form at full spreading rates less than about 12 mm yr(-1), though their characteristics are commonly found at rates up to approximately 20 mm yr(-1). The ultraslow-spreading ridges consist of linked magmatic and amagmatic accretionary ridge segments. The amagmatic segments are a previously unrecognized class of accretionary plate boundary structure and can assume any orientation, with angles relative to the spreading direction ranging from orthogonal to acute. These amagmatic segments sometimes coexist with magmatic ridge segments for millions of years to form stable plate boundaries, or may displace or be displaced by transforms and magmatic ridge segments as spreading rate, mantle thermal structure and ridge geometry change.


NCCN Guidelines® Insights: Cervical Cancer, Version 1.2024.
The NCCN Guidelines for Cervical Cancer provide recommendations for all aspects of management for cervical cancer, including the diagnostic workup, staging, pathology, and treatment. The guidelines also include details on histopathologic classification of cervical cancer regarding diagnostic features, molecular profiles, and clinical outcomes. The treatment landscape of advanced cervical cancer is evolving constantly. These NCCN Guidelines Insights provide a summary of recent updates regarding the systemic therapy recommendations for recurrent or metastatic disease.


Those Left Behind From Voluntary Medical Home Reforms in Ontario, Canada.
Health systems are transitioning patients to medical homes to improve health outcomes and reduce cost. We sought to understand the characteristics and quality of care for patients who did and did not participate in the voluntary transition to medical homes. We used administrative data for diabetes monitoring and cancer screening to compare services received by patients attached to a medical home (n = 10,785,687) with services received by those seeing a fee-for-service physician (n = 1,321,800) in Ontario, Canada, on March 31, 2011. We used Poisson regression to examine associations in 2011 after adjustment for patient factors and also assessed changes in outcomes between 2001 and 2011. Patients attached to a fee-for-service physician were more likely to be immigrants and live in a low-income neighborhood and urban area. They were less likely to receive recommended testing for diabetes (25% vs 34%; adjusted relative risk [RR] = 0.74; 95% CI, 0.73-0.75) and less likely to receive screening for cervical (52% vs 66%; adjusted RR = 0.79; 95% CI, 0.79-0.79), breast (58% vs 73%; adjusted RR = 0.80; 95% CI, 0.80-0.81), and colorectal cancer (44% vs 62%; adjusted RR = 0.72; 95% CI, 0.71-0.72) compared with patients attached to a medical home physician in 2011. These differences in quality of care preceded medical home reforms. Patients left behind from medical home reforms are more likely to be poor, urban, and new immigrants and receive lower quality care. Strategies are needed to reach out to these patients and their physicians to reduce gaps in care.


SICKO. Development of a Multi-Disciplinary Training-Concept for Paediatric Oncology.
Background Treating children with cancer requires multiple different skills. For the healthcare personnel (HCP) in Germany the practice of ongoing training to improve professional skills is almost non-existent. Therefore, we developed a programme called 'SICKO' to support HCPs skills and attitudes by means of a multidisciplinary workshop. Methods Following a qualitative analysis, we then designed a modular (3 day) workshop. During day one (8 h) participants learn practical skills, the fundamentals of chemotherapy, and effective communication skills. Workshop day 2 (8 h) includes education regarding the complications of cancer therapy (e. g. tumour-lysis syndrome, delayed methotrexate excretion), and their management. Topics during day 3 (8 h) include 'breaking bad news', conflict management in the team, infusion-related complications and 'crew resource management' (CRM). Results 43 nurses and 33 physicians participated between 2013 and 2015. All participants highly recommend the workshop. Participants felt that knowledge increased significantly after workshops and were more confident regarding challenging communications. Discussion/Conclusions Although long-term effects have not yet been evaluated, 'SICKO' offers the opportunity for HCP to train and experience simulated day-to-day challenges in the field of paediatric oncology.


Deciphering the Mechanism of Crystallization of UiO-66 Metal-Organic Framework.
Zirconium-containing metal-organic framework (MOF) with UiO-66 topology is an extremely versatile material, which finds applications beyond gas separation and catalysis. However, after more than 10 years after the first reports introducing this MOF, understanding of the molecular-level mechanism of its nucleation and growth is still lacking. By means of in situ time-resolved high-resolution mass spectrometry, Zr K-edge X-ray absorption spectroscopy, magic-angle spinning nuclear magnetic resonance spectroscopy, and X-ray diffraction it is showed that the nucleation of UiO-66 occurs via a solution-mediated hydrolysis of zirconium chloroterephthalates, whose formation appears to be autocatalytic. Zirconium-oxo nodes form directly and rapidly during the synthesis, the formation of pre-formed clusters and stable non-stoichiometric intermediates are not observed. The nuclei of UiO-66 possess identical to the crystals local environment, however, they lack long-range order, which is gained during the crystallization. Crystal growth is the rate-determining step, while fast nucleation controls the formation of the small crystals of UiO-66 with a narrow size distribution of about 200 nanometers.


Are Biosimilars the Future of Oncology and Haematology?
Biological drugs are vital but often high-cost components of cancer treatment. Several biosimilar versions of these drugs have been approved in Europe and/or the USA, with many more in development. However, there is some disconnect between the biosimilars that are approved for use and those accessible in clinical practice, with availability impacted by factors including patent litigation and complex healthcare insurance policies, particularly in the USA. Provided the barriers to widespread uptake can be overcome, biosimilars offer potential benefits including cost savings and improved patient access versus the reference product (RP). This article provides an up-to-date and focused perspective on the development and use of biosimilars in the haemato-oncology setting. European and US regulatory pathways governing biosimilar licensing demand that there are no clinically meaningful differences between a biosimilar and its RP. Pathways are rigorously enforced and involve comprehensive non-clinical evaluations and clinical trials in selected indications to establish the equivalence or non-inferiority of efficacy, and the comparability of safety, of the biosimilar versus its RP. 'Indication extrapolation' is only permitted if scientifically justifiable considering mechanism(s) of action, pharmacokinetics, immunogenicity and safety in relevant patient populations. Switching treatment from RP to biosimilar is supported by most available data, predominantly from indications other than cancer, and post-marketing pharmacovigilance programmes are warranted. Notably, the potential benefits of biosimilar cancer treatment may extend beyond direct cost savings: for example, the availability of biosimilars of common regimen components may help incentivise the evaluation and/or clinical use of new treatment approaches and novel drugs.


Screening for oxygen intolerance in U.S. Navy divers.
All U.S. Navy diving candidates are screened for their tolerance to hyperbaric oxygen by taking an oxygen tolerance test (OTT). During a recent experimental oxygen dive series at the U.S. Navy Experimental Diving Unit, three divers were noted to be reproducibly oxygen sensitive. These three divers were then given additional OTTs to see if any evidence of central nervous system oxygen toxicity would be detected by these multiple tests. The additional OTTs did not produce any signs or symptoms of oxygen toxicity in these already proven susceptible divers. A subsequent review of the records of the Naval Safety Center yielded a total of 1347 OTTs from 1 January 1972 to 31 December 1981. A review of diving accidents reported during this period revealed that 26 episodes of oxygen toxicity were noted during OTTs for a derived failure rate of 1.9%. Analysis of oxygen toxicity episodes encountered during operational Navy diving for this period found that 9 episodes of nonconvulsive oxygen toxicity were seen in mixed gas diving and 3 episodes of nonconvulsive oxygen toxicity were noted in closed circuit oxygen diving. Conclusions from this paper are: Screening for oxygen intolerance is complicated by intraindividual variation in oxygen tolerance; U.S. Navy diving using 100% oxygen during the period studied has had an acceptable safety record according to the data on record at the Naval Safety Center; the OTT as currently administered by the U.S. Navy does not identify all individuals who are relatively susceptible to oxygen toxicity; those individuals who do fail the OTT are unusually susceptible to oxygen toxicity; and because of the need to continue to identify these unusually susceptible individuals, the OTT should continue to be administered to U.S. Navy diver candidates.


Longitudinal study of giardiasis in three day care centres of Havana City.
The prevalence, incidence and reinfection of giardiasis were studied in 365 children attending three day care centres (DCCs) in Havana City. Three stool samples were obtained from each child every 6 months during an 18-month period. We identified three distinct groups of children according to their patterns of infection. In the largest group (51%) children were never found infected. In the second group, they were found infected once or twice during the study period, and in the third and the smallest group (9%) they were found infected in most or all the study periods. This last group seems to be children 'predisposed' to Giardia lamblia infection. The prevalence of giardiasis (20%) remained almost constant throughout the study period. The incidence declined from 16 to 11%, and reinfection increased from 36 to 49%. All the children had normal nutritional status and the only clinical manifestation that correlated strongly with the frequency of cross-sectional surveys positive to Giardia was the number of diarrhoeal episodes recorded during the last 6 months of the study period. Further studies will be necessary to ascertain the causes that determine the 'predisposition' to giardiasis in children.


Protein crystal growth in microgravity.
Protein crystallography is a powerful method for determining the three-dimensional structures of biological macromolecules. Although new methods, such as two-dimensional NMR, have demonstrated promise for determining the structures of small proteins and nucleic acids, the complete atomic arrangements within large proteins can only be determined at present using crystallographic techniques. Such crystallographic studies have been of major importance for establishing structure/function relationships that are fundamental to understanding how enzymes, nucleic acids, and other macromolecules function in biological systems. More recently, crystallographic studies of proteins have become of considerable practical interest within the pharmaceutical and biotechnology industries, as promising tools in drug design and in protein engineering.


Role of glucocorticoid signaling in urothelial tumorigenesis: Inhibition by prednisone presumably through inducing glucocorticoid receptor transrepression.
Glucocorticoids, including dexamethasone (DEX) and prednisone (PRED), have been prescribed in patients with neoplastic disease as cytotoxic agents or comedications. Nonetheless, it remains uncertain whether they have an impact on the development of bladder cancer. We, therefore, assessed the functional role of the glucocorticoid-mediated glucocorticoid receptor (GR) signaling in urothelial tumorigenesis. Tumor formation was significantly delayed in xenograft-bearing mice with implantation of control bladder cancer UMUC3 cells or nonneoplastic urothelial SVHUC cells undergoing malignant transformation induced by a chemical carcinogen 3-methylcholanthrene (MCA), compared with respective GR knockdown xenografts. Using the in vitro system with MCA-SVHUC cells, we screened 11 GR ligands, including DEX, and found significant inhibitory effects of PRED on their neoplastic transformation. The effects of PRED were restored by a GR antagonist RU486 in GR-positive MCA-SVHUC cells, while PRED failed to inhibit the neoplastic transformation of GR knockdown cells. Significant decreases in the expression levels of oncogenes (c-Fos/c-Jun) and significant increases in those of a tumor suppressor UGT1A were seen in MCA-SVHUC-control cells (vs GR-short hairpin RNA) or PRED-treated MCA-SVHUC-control cells (vs mock). In addition, N-butyl-N-(4-hydroxybutyl) nitrosamine induced bladder cancer in all of eight mock-treated mice vs seven (87.5%) of DEX-treated (P = .302) or four (50%) of PRED-treated (P = .021) animals. Finally, DEX was found to considerably induce both transactivation (activation of glucocorticoid-response element mediated transcription and expression of its targets) and transrepression (suppression of nuclear factor-kappa B transactivation and expression of its regulated genes) of GR in SVHUC cells, while PRED more selectively induced GR transrepression. These findings suggest that PRED could prevent urothelial tumorigenesis presumably via inducing GR transrepression.


Rapid diagnosis of adenoviral conjunctivitis by PCR and restriction fragment length polymorphism analysis.
To detect and identify adenovirus (Ad), we used a combination of PCR and restriction fragment length polymorphism (RFLP) analysis. Nested PCR with two primer sets that hybridize to the conserved region for hexon proteins of 14 prototypes of Ad, Ad serotype 1 (Ad1) to Ad8, -11, -14, -19, -37, -40, and -41, amplified a 956-bp DNA fragment. The amplified fragments from the 14 prototypes were completely differentiated with a combination of three restriction endonucleases, EcoT14I, HaeIII, and HintI. We applied this new method for 127 samples of conjunctival scrapings from patients with conjunctivitis and compared the results with those obtained with the combination of culture isolation and a neutralization test (NT). PCR gave a positive result in 69 of 127 cases (54.3%), while only 61 of the 127 samples (48.0%) tested positive by culture isolation. Compared with isolation, the PCR method had a sensitivity of 100% (61 of 61). Positive PCR samples were further classified as Ad37 (59.5%), -3(31.9%), -11 (4.3%), -8 (2.9%), and -4 (1.4%) by PCR-RFLP analysis. Of eight samples that were PCR positive and culture isolation negative, six were Ad37 and two were Ad8 by PCR-RFLP analysis. These differentiations of isolation-positive samples were identical to the results obtained by the NT. It took only 3 days to detect and identify Ad by PCR-RFLP analysis, whereas it took at least 3 weeks by culture isolation and NT. Our newly developed method of detecting and typing human Ad by PCR-RFLP analysis is more sensitive, accurate, and rapid than the conventional method of culture isolation and an NT.


Leucocyte and platelet adhesion in different layers of the small bowel during experimental total warm ischaemia and reperfusion.
Ischaemia and reperfusion (IR) of the small bowel is involved in many clinical conditions. A key component in IR-induced tissue damage is microvascular dysfunction. The aim was to investigate the role of leucocytes and platelets in capillary flow impediment and tissue damage. Anaesthetized rats were subjected to 30 min warm ischaemia of the small bowel, followed by 1 h reperfusion. To elucidate the influence of leucocytes on platelet adhesion, leucocyte-vessel wall interactions induced by IR were prevented by anti-platelet activating factor (PAF) or anti-intercellular adhesion molecule (ICAM)-1. Intravital videomicroscopy was performed and tissue injury was evaluated histologically. In submucosal venules, IR induced an increase in the median number of interacting leucocytes from 3 to 10 and 20 leucocytes per 100-microm venule segment after 10 and 60 min reperfusion respectively. Anti-PAF or anti-ICAM-1 completely attenuated this increase, resulting in an eightfold improvement in submucosal capillary flow and reduced tissue injury. Shedding of villi no longer occurred. Platelet-vessel wall interactions occurred particularly in submucosal venules, but were not affected by anti-PAF or anti-ICAM-1. Small bowel IR initiated an inflammatory and thrombotic response in the submucosal layer only. Attenuation of leucocyte adhesion improved submucosal capillary perfusion, preventing shedding of mucosal villi.


Increased endothelin-induced Ca2+ signaling, tyrosine phosphorylation, and coronary artery disease in diabetic dyslipidemic Swine are prevented by atorvastatin.
Endothelin-1 (ET-1) signaling mechanisms have been implicated in the pathogenesis of excess coronary artery disease in diabetic dyslipidemia. We hypothesized that in diabetic dyslipidemia ET-1-induced coronary smooth muscle calcium (Ca2+m) and tyrosine phosphorylation would be increased, and the lipid lowering agent, atorvastatin, would inhibit these increases. Male Yucatan miniature swine groups were treated for 20 weeks: normal low-fat fed control, high-fat/cholesterol fed (hyperlipidemic), hyperlipidemic made diabetic with alloxan (diabetic dyslipidemic), and diabetic dyslipidemic treated with atorvastatin (atorvastatin-treated). Blood glucose values were 5-fold greater in diabetic dyslipidemic and atorvastatin-treated versus control and hyperlipidemic. Total and low-density lipoprotein (LDL) plasma cholesterol in hyperlipidemic, diabetic dyslipidemic, and atorvastatin-treated were approximately 5-fold greater than control. Intravascular ultrasound detectable coronary disease and hypertriglyceridemia were only observed in diabetic dyslipidemic and were abolished by atorvastatin. In freshly isolated cells, the Ca2+m response to ET-1 in diabetic dyslipidemic was greater than in control, hyperlipidemic, and atorvastatin-treated groups. Selective ET-1 receptor antagonists showed in the control group that the ETB subtype inhibits ETA regulation of Ca2+m. There was almost a complete switch of receptor subtype regulation of Ca2+m from largely ETA in control to an increased inhibitory interaction between ETA and ETB in hyperlipidemic and diabetic dyslipidemic groups, such that neither ETA nor ETB antagonist alone could block the ET-1-induced Ca2+m response. The inhibitory interaction was attenuated in the atorvastatin-treated group. In single cells, basal and ET-1-induced tyrosine phosphorylation in diabetic dyslipidemic were more than 3- and 6-fold greater, respectively, than in control, hyperlipidemic, and atorvastatin-treated. Attenuation by atorvastatin of coronary disease and ET-1-induced Ca2+m and tyrosine phosphorylation signaling with no change in cholesterol provides strong evidence for direct actions of atorvastatin and/or triglycerides on the vascular wall.


Facile preparation of gadolinium(iii) chelates functionalized carbon quantum dot-based contrast agent for magnetic resonance/fluorescence multimodal imaging.
Suffering from high inherent toxicity generated from heavy metals in semiconductor quantum dots and a complex preparation process, the medical application of conventional gadolinium(iii)/semiconductor quantum dot-based magnetic resonance/fluorescence multimodal imaging (MRI/FI) contrast agent has been limited. In this paper, a new kind of biocompatible carbon quantum dot functionalized by gadolinium(iii) chelates (named as Gd(iii)/CQDs) was obtained through one-step pyrolysis of gadopentetate monomeglumine, which provides simultaneously a carbon source and a gadolinium(iii) source. The Gd(iii)/CQDs show good magnetic resonance response and photoluminescence properties with longitudinal relaxation rate and quantum yield of 5.5-6.4 mM L-1 S-1 [Gd(iii)%, 0.5 mM] and 2.6-8.9%, respectively. The Gd(iii)/CQDs with diameters in the range of 2-3 nm can be well dispersed in deionized water, phosphate-buffered saline buffer solution or culture medium to form a stable dispersion, which with low cytotoxicity can penetrate into cells and show green fluorescence (excitation at 488 nm), presenting a great potential to be used as an MRI/FI contrast agent.


Global epidemiology of nonalcoholic fatty liver disease-Meta-analytic assessment of prevalence, incidence, and outcomes.
Nonalcoholic fatty liver disease (NAFLD) is a major cause of liver disease worldwide. We estimated the global prevalence, incidence, progression, and outcomes of NAFLD and nonalcoholic steatohepatitis (NASH). PubMed/MEDLINE were searched from 1989 to 2015 for terms involving epidemiology and progression of NAFLD. Exclusions included selected groups (studies that exclusively enrolled morbidly obese or diabetics or pediatric) and no data on alcohol consumption or other liver diseases. Incidence of hepatocellular carcinoma (HCC), cirrhosis, overall mortality, and liver-related mortality were determined. NASH required histological diagnosis. All studies were reviewed by three independent investigators. Analysis was stratified by region, diagnostic technique, biopsy indication, and study population. We used random-effects models to provide point estimates (95% confidence interval [CI]) of prevalence, incidence, mortality and incidence rate ratios, and metaregression with subgroup analysis to account for heterogeneity. Of 729 studies, 86 were included with a sample size of 8,515,431 from 22 countries. Global prevalence of NAFLD is 25.24% (95% CI: 22.10-28.65) with highest prevalence in the Middle East and South America and lowest in Africa. Metabolic comorbidities associated with NAFLD included obesity (51.34%; 95% CI: 41.38-61.20), type 2 diabetes (22.51%; 95% CI: 17.92-27.89), hyperlipidemia (69.16%; 95% CI: 49.91-83.46%), hypertension (39.34%; 95% CI: 33.15-45.88), and metabolic syndrome (42.54%; 95% CI: 30.06-56.05). Fibrosis progression proportion, and mean annual rate of progression in NASH were 40.76% (95% CI: 34.69-47.13) and 0.09 (95% CI: 0.06-0.12). HCC incidence among NAFLD patients was 0.44 per 1,000 person-years (range, 0.29-0.66). Liver-specific mortality and overall mortality among NAFLD and NASH were 0.77 per 1,000 (range, 0.33-1.77) and 11.77 per 1,000 person-years (range, 7.10-19.53) and 15.44 per 1,000 (range, 11.72-20.34) and 25.56 per 1,000 person-years (range, 6.29-103.80). Incidence risk ratios for liver-specific and overall mortality for NAFLD were 1.94 (range, 1.28-2.92) and 1.05 (range, 0.70-1.56). As the global epidemic of obesity fuels metabolic conditions, the clinical and economic burden of NAFLD will become enormous. (Hepatology 2016;64:73-84).


The natural history of multiple sclerosis.
The natural history of chronic diseases is generally poorly understood. Multiple sclerosis is one of the best-studied diseases and we outline here some of the information obtained from 25,000 patient-years of observation. This population-based cohort received almost no treatment and certainly none known to alter the long-term natural course of the illness. There is a possibility that such data will prove useful in new generation clinical trials which are observational in nature.


[Transverse presentation due to coprostasis].
A multiparous woman was admitted at 32 weeks to evaluate the reason for a persistency in transverse lie. Ultrasound examination at 25 and 32 weeks revealed a circular echogenic structure near the cervix. It was considered to be a fibroid. At rectovaginal examination a compact faecal mass was identified. The woman never had complained of abnormal bowel function: however, daily diarrhoea had occurred since 3 months gestational age. Several times it was necessary to apply oil rectally for softening, enabling digital faecal extraction; phosphate enemas and warm water enemas led to considerable faeces production. Near term a healthy daughter was delivered vaginally. Chronic constipation was diagnosed, worsened by pregnancy and an inadequate diet.


The delta beta crossover region in Lepore boston hemoglobinopathy is restricted to a 59 base pairs region around the 5' splice junction of the large globin gene intervening sequence.
Genomic DNA from a hemoglobin (Hb) Lepore Boston (delta 87 Gln beta 116 His) homozygote of Southern Italian origin has been studied in order to map the fusion point between the delta and beta genes. An Ava II restriction endonuclease recognition sequence, located 12 base pairs (bp) downstream from the 5' end of the beta gene large intervening sequence, has been taken as marker of the beta-like portion of the fusion gene. This site was present even in the delta beta gene, allowing the localization of the crossover area to a 59-bp region extending from the first nucleotide of the Leu codon in position 88 to the 11th nucleotide of the large intervening sequence. The analysis of the DNA restriction polymorphisms in the gamma delta beta globin gene region provides evidence that a single mutational event originated the Lepore delta beta genes, at least in the Italian population.


Similar genomic patterns of clinical infective endocarditis and oral isolates of Streptococcus sanguinis and Streptococcus gordonii.
Streptococcus gordonii and Streptococcus sanguinis belong to the Mitis group streptococci, which mostly are commensals in the human oral cavity. Though they are oral commensals, they can escape their niche and cause infective endocarditis, a severe infection with high mortality. Several virulence factors important for the development of infective endocarditis have been described in these two species. However, the background for how the commensal bacteria, in some cases, become pathogenic is still not known. To gain a greater understanding of the mechanisms of the pathogenic potential, we performed a comparative analysis of 38 blood culture strains, S. sanguinis (n = 20) and S. gordonii (n = 18) from patients with verified infective endocarditis, along with 21 publicly available oral isolates from healthy individuals, S. sanguinis (n = 12) and S. gordonii (n = 9). Using whole genome sequencing data of the 59 streptococci genomes, functional profiles were constructed, using protein domain predictions based on the translated genes. These functional profiles were used for clustering, phylogenetics and machine learning. A clear separation could be made between the two species. No clear differences between oral isolates and clinical infective endocarditis isolates were found in any of the 675 translated core-genes. Additionally, random forest-based machine learning and clustering of the pan-genome data as well as amino acid variations in the core-genome could not separate the clinical and oral isolates. A total of 151 different virulence genes was identified in the 59 genomes. Among these homologs of genes important for adhesion and evasion of the immune system were found in all of the strains. Based on the functional profiles and virulence gene content of the genomes, we believe that all analysed strains had the ability to become pathogenic.


Evaluation of tebufenozide carry-over and residual effects on spruce budworm (Lepidoptera: Tortricidae).
Laboratory and field studies investigated carry-over effects of tebufenozide on spruce budworm, Choristoneura fumiferana (Clemens). In the laboratory, third and fourth instars were fed either sublethal doses of tebufenozide (10 ppm) or water onAbies balsamea (L.) Mill. needles, reared to adulthood and allowed to oviposit on laying surfaces 1 or 14 d after being sprayed with water or tebufenozide concentrations of 17.5, 35.0, and 70.0 g/liter. Percentage adult emergence and sex ratio were not affected by larval ingestion of the tebufenozide. Also, the mean number of eggs laid on untreated wax paper by moths reared from tebufenozide-treated larvae was similar to the controls. Hence, tebufenozide did not inhibit carry-over effects on treated larvae. Oviposition on tebufenozide-treated wax paper by moths reared from untreated larvae was affected by both the substrate concentration and the age of the treatment residue. When offered treated and untreated laying surfaces simultaneously, C. fumiferana did not show a preference. However, significantly fewer eggs were laid on both laying surfaces by fewer females than when tebufenozide was absent. Residual tebufenozide on wax paper did not affect egg hatch but topical applications were toxic to eggs. Field studies appear to corroborate laboratory results and suggest that although the ingestion of tebufenozide by larval spruce budworm might not impair adult reproduction, the insecticide's presence in the environment could inhibit oviposition. This inhibition was considered to be a primary factor in tebufenozide's multi-year effects against spruce budworm populations.


Fatal hemorrhage complicating carcinoma of the esophagus. Report of four cases.
Four cases of esophageal carcinoma complicated by fatal hemorrhage are reported. All four patients had recently completed radiation therapy. An aortoesophageal fistula was present in two cases; fibrinoid necrosis of the esophageal arteries was present in the other two. The esophageal tumor was localized in two cases and had disappeared in one case. In one patient it had metastasized widely. Ninety-nine other reports of esophageal cancer and fatal hemorrhage are reviewed from the literature. Aortoesophageal fistula was the cause of hemorrhage in 78 cases. Occlusion of the vasa vasorum by thrombosis, inflammation, neoplastic cells or radiation injury appears to be the cause of aortic necrosis and fistula formation. Prompt surgical approach, if possible, should be used to control hemorrhage, as the primary tumor may be localized to the esophagus only.


Role of cytosol in the stimulation of RNA transport in vitro during cardiac hypertrophy in rats.
The 100,000 g supernatant isolated from hypertrophic hearts on fractionation by (NH4)2SO4 and DEAE-cellulose chromatography showed an enhanced RNA-transport activity when incubated with isolated nuclei from sham-operated hearts in vitro. Proteins of Mr 73,000, 68,000, 43,000 and 32,000 are enriched in the DEAE-cellulose fractions exhibiting maximal transport activity, and they are phosphorylatable. Pretreatment of the cytosol with antibodies to the Mr-68,000 and -32,000 proteins decreases the transport activity of the cytosol from 14% to 4.25%. Proteins of Mr 73,000, 68,000, 43,000 and 32,000 are translocated from the cytosol to the nuclear envelope under conditions of RNA transport in vitro. Our results here suggest that at least two of these proteins, those of Mr 68,000 and 32,000, play an indispensible role in the nucleocytoplasmic RNA transport in vitro. By making use of a specific myosin heavy-chain B-gene probe and hybridization, we have also shown the effect of cytosol on the transport of myosin heavy-chain mRNA from nucleus to cytosol.


Primary contact sites in intrinsically unstructured proteins: the case of calpastatin and microtubule-associated protein 2.
Intrinsically unstructured proteins (IUPs) exist in a disordered conformational state, often considered to be equivalent with the random-coil structure. We challenge this simplifying view by limited proteolysis, circular dichroism (CD) spectroscopy, and solid-state (1)H NMR, to show short- and long-range structural organization in two IUPs, the first inhibitory domain of calpastatin (CSD1) and microtubule-associated protein 2c (MAP2c). Proteases of either narrow (trypsin, chymotrypsin, and plasmin) or broad (subtilisin and proteinase K) substrate specificity, applied at very low concentrations, preferentially cleaved both proteins in regions, i.e., subdomains A, B, and C in CSD1 and the proline-rich region (PRR) in MAP2c, that are destined to form contacts with their targets. For CSD1, nonadditivity of the CD spectra of its two halves and suboptimal hydration of the full-length protein measured by solid-state NMR demonstrate that long-range tertiary interactions provide the structural background of this structural feature. In MAP2c, such tertiary interactions are absent, which points to the importance of local structural constraints. In fact, urea and temperature dependence of the CD spectrum of its PRR reveals the presence of the extended and rather stiff polyproline II helix conformation that keeps the interaction site exposed. These data suggest that functionally significant residual structure exists in both of these IUPs. This structure, manifest as either transient local and/or global organization, ensures the spatial exposure of short contact segments on the surface. Pertinent data from other IUPs suggest that the presence of such recognition motifs may be a general feature of disordered proteins. To emphasize the possible importance of this structural trait, we propose that these motifs be called primary contact sites in IUPs.


Clarithromycin vs ciprofloxacin as adjuncts to rifampicin and ethambutol in treating opportunist mycobacterial lung diseases and an assessment of Mycobacterium vaccae immunotherapy.
The mainstays of treatment for pulmonary disease caused by opportunist mycobacteria are rifampicin (R) and ethambutol (E). The role of macrolides, quinolones and immunotherapy with Mycobacterium vaccae is not clear. A trial was undertaken to compare clarithromycin (Clari) and ciprofloxacin (Cipro) as third drugs added to [corrected] 2 years of treatment with R and E for pulmonary disease caused by M avium-intracellulare (MAC), M malmoense and M xenopi (REClari and RECipro). An optional comparison of immunotherapy with M vaccae vs no immunotherapy was also performed. Progress was monitored annually during the 2 years of treatment and for 3 years thereafter. If the patient was not improving at 1 year the regimen was supplemented by the addition of the drug not received in the original allocation of treatment. 371 patients (186 REClari, 185 RECipro) entered the study (170 MAC, 167 M malmoense, 34 M xenopi). All-cause mortality was high for both groups (44% REClari, 43% RECipro); for MAC it was higher with REClari than with RECipro (48% vs 29%) but for M malmoense (42% vs 56%) and M xenopi (29% vs 47%) it was higher with RECipro (p = 0.006). 3% died from their mycobacterial disease (REClari = RECipro). At the end of treatment, 4% of REClari and 10% of RECipro patients still had positive cultures. Among those with negative cultures at the end of treatment, 6% of the REClari group and 4% of the RECipro group had relapsed. At 5 years 30% of the REClari group were known to have completed treatment as allocated and to be alive and cured compared with 21% of the RECipro group (p = 0.04), but this difference was principally due to those with M malmoense (REClari 38%, RECipro 20%). Patients with MAC or M xenopi were more likely to have a poor outcome than those with M malmoense (p = 0.004), with no difference between REClari and RECipro. Overall, 20% in each group were unable to tolerate the regimen allocated, Cipro being associated with more unwanted effects than Clari (16% vs 9%, p = 0.05). No significant differences in outcomes were found between M vaccae-treated patients and those not treated with M vaccae immunotherapy. Considering all three species together, there were no differences in outcome between the REClari and RECipro groups. Immunotherapy did not improve outcome. New therapies, optimised management of co-morbid conditions and a more holistic approach must be explored in the hope of improving outcome.


InflammAging at Ocular Surface: Clinical and Biomolecular Analyses in Healthy Volunteers.
To assess the ocular surface in volunteers who consider themselves as healthy, in order to evaluate how para-inflammatory mechanisms fail with age, and thus investigate the phenomenon of "InflammAging." In this observational prospective cohort study, volunteers were categorized into three groups according to age: young (19-40 years), middle-aged (41-60 years), and older adults (61-93 years). Clinical assessments included tear breakup time (T-BUT) and Schirmer test type I. Dry eye symptoms were evaluated by the Ocular Surface Disease Index (OSDI) questionnaire. Conjunctival mRNA and protein expression of intercellular adhesion molecule-1 (ICAM-1), MUC5AC, and IL-8 were measured by real-time PCR and immunofluorescence. A total of 82 volunteers (38 males and 44 females) were enrolled. T-BUT decreased significantly with increasing age (young: 11.13 ± 0.18 seconds; middle-aged: 10.83 ± 0.56 seconds; older: 9.00 ± 1.00 seconds, P < 0.05). Schirmer test values decreased significantly with age (young: 20.6 ± 1.0 mm; middle-aged: 19.2 ± 1.2 mm; older: 16.0 ± 1.1 mm, P < 0.05). OSDI scores increased with age in both groups, but they were substantially higher in women. Conjunctival expression of inflammatory markers ICAM-1, IL-8, and MUC5AC increased with age. Clinical signs, symptoms, and biomarkers of chronic inflammation increased with age in a cohort of volunteers who considered themselves healthy, indicating an age-related progressive impairment of ocular surface system function.


Incidence of testicular germ-cell malignancies in England and Wales: trends in children compared with adults.
The incidence of testicular cancer has been increasing markedly in most industrialised countries. This rise is known to have affected young adults, but it is less clear whether it has affected other age groups, particularly children. We used data from the National Cancer Registry file at the Office of National Statistics (ONS) and the National Registry of Childhood Tumours to examine trends in testicular germ-cell malignancies overall in England and Wales from 1962 to 1990 and in children from 1962 to 1995. The incidence of testicular cancer at all ages rose by 3.4% (95% CI 3.3-3.6%) per annum from 1962 to 1990. A similar rise in the incidence of germ-cell malignancies occurred during the years for which histological information was available in the ONS files, 1971-1989 (3.4%; 3.1-3.6%), to which both seminomas and non-seminomas contributed equally. The incidence of non-seminomas in adults rose in men under age 55 years and declined in older men, whereas there were increases in the incidence of seminomas in both young and older men. Cohort analysis at young ages showed a marked rise in the risk of germ-cell malignancies up to the cohort born in 1955-1959 but no further rise for those born subsequently. The rise in the incidence of these tumours in young adults was paralleled by a similar trend, although less marked, in children aged under 15 years (1.3% per annum; 0.2-2.5%). The increase in risk for children in this very large data set alongside the rise in young adults is compatible with the hypothesis that childhood and adult testicular germ-cell malignancies may have some common risk factors, presumably pre-natal.


[Malformations of the central nervous system: analysis of 157 pediatric autopsies].
The malformations of the central nervous system affect about 5 to 10 children per 1000 births. We studied the central nervous system malformations in 5837 pediatric autopsies performed in the Sector of Anatomic Pathology, Hospital de Clínicas-UFPR, between 1960 and 1995. There were 157 central nervous system malformations (2.69%), the commonest were neural tube defects (61%): 47 cases of anencephaly and 45 cases classified in the group of myeloencephaloceles. The anomalies of the prosencephalic evagination corresponded to 8% of all central nervous system malformations, with seven cases of holoprosencephaly. Posterior fossa malformations occurred in 3%, with three cases of Arnold-Chiari. In the present study, the mortality rate due to central nervous system malformations was higher in the neonatal period.


Human paraoxonases (PON1, PON2, and PON3) are lactonases with overlapping and distinct substrate specificities.
The paraoxonase (PON) gene family in humans has three members, PON1, PON2, and PON3. Their physiological role(s) and natural substrates are uncertain. We developed a baculovirus-mediated expression system, suitable for all three human PONs, and optimized procedures for their purification. The recombinant PONs are glycosylated with high-mannose-type sugars, which are important for protein stability but are not essential for their enzymatic activities. Enzymatic characterization of the purified PONs has revealed them to be lactonases/lactonizing enzymes, with some overlapping substrates (e.g., aromatic lactones), but also to have distinctive substrate specificities. All three PONs metabolized very efficiently 5-hydroxy-eicosatetraenoic acid 1,5-lactone and 4-hydroxy-docosahexaenoic acid, which are products of both enzymatic and nonenzymatic oxidation of arachidonic acid and docosahexaenoic acid, respectively, and may represent the PONs' endogenous substrates. Organophosphates are hydrolyzed almost exclusively by PON1, whereas bulky drug substrates such as lovastatin and spironolactone are hydrolyzed only by PON3. Of special interest is the ability of the human PONs, especially PON2, to hydrolyze and thereby inactivate N-acyl-homoserine lactones, which are quorum-sensing signals of pathogenic bacteria. None of the recombinant PONs protected low density lipoprotein against copper-induced oxidation in vitro.


Hypertonic saline maintains coagulofibrinolytic homeostasis following moderate‑to‑severe traumatic brain injury by regulating monocyte phenotype via expression of lncRNAs.
Traumatic brain injury (TBI) is the most common cause of death and permanent disability in people aged <45, and is associated with secondary brain injury and bleed progression, resulting in increased morbidity and mortality. TBI may also induce innate host defense responses characterized by activation of resident microglia and astrocytes, brain microvascular endothelial cells and peripheral blood monocytes. In the present study, 34 patients with moderate‑to‑severe traumatic brain injury were randomly divided into two groups, including a 7.5% hypertonic saline (HS) treatment group (4 ml/kg) and 3% HS treatment group (4 ml/kg). The results demonstrated that treatment with 7.5% HS decreased the intracranial pressure and improved coagulofibrinolytic homeostasis. Analysis of the monocyte subsets revealed significant reduction in the proportion of cluster of differentiation (CD)14++CD16+ circulating inflammatory monocytes in the 7.5% HS group. In addition, 7.5% HS treatment downregulated the expression of long non‑coding (lnc) RNA2448‑11 and lncRNA1403 in the peripheral blood mononuclear cells of patients with TBI. Using reverse transcription‑quantitative polymerase chain reaction, it was determined that 7.5% HS regulated the expression of tumor necrosis factor‑α, interleukin‑1β, transforming growth factor‑β and thrombomodulin, which are the target genes of lncRNA2448‑11 and lncRNA1403. These results indicated that 7.5% HS improved the intracranial pressure and coagulofibrinolytic homeostasis by modulating the phenotype of monocytes through lncRNA2448‑11 and lncRNA1403. These findings provided evidence that initial resuscitation with HS imparts functional changes to inflammatory cells following TBI, thereby reducing potential neuroinflammatory events associated with secondary brain injury.


Risk-managed production of bioactive recombinant proteins using a novel plant virus vector with a helper plant to complement viral systemic movement.
A plant viral vector has the potential to efficiently produce recombinant proteins at a low cost in a short period. Although recombinant proteins can be also produced by transgenic plants, a plant viral vector, if available, may be more convenient when urgent scale-up in production is needed. However, it is difficult to use a viral vector in open fields because of the risk of escape to the environment. In this study, we constructed a novel viral vector system using a movement-defective Cucumber mosaic virus (CMV) vector, which is theoretically localized in the inoculated cells but infects systemically only with the aid of the transgenic helper plant that complements viral movement, diminishing the risk of viral proliferation. Interestingly, the helper plant systemically infected with the vector gave strong cross-protection against challenge inoculation with wild-type CMVs. Using CMV strains belonging to two discrete CMV groups (subgroups I and II), we also improved the system to prevent recombination between the vector and the transgene transcript in the helper plant. We here demonstrate the expression of an anti-dioxin single chain variable fragment (DxscFv) and interleukin-1 receptor antagonist (IL1-Ra) in Nicotiana benthamiana by this viral vector confinement system, which is applicable for many useful high-quality recombinant proteins.


How minority members' perceptions of majority members' acculturation preferences shape minority members' own acculturation preferences: evidence from Chile.
Two survey studies were conducted in Chile with members of the indigenous minority group Mapuche (Ns = 566; 394). The aim was to find predictors of minority members' acculturation preferences, especially integration. It was hypothesized that minority members' preferences would depend on their perceptions of what majority members want. Specifically, it was predicted that a perception that majority members want minority members to maintain their original culture would be associated with a greater desire for culture maintenance among minority participants. Further, it was predicted that a perception that majority members want intergroup contact would be associated with a greater desire for contact among minority participants. Finally, it was predicted that a perception that majority members are in favour of both culture maintenance and contact (i.e., integration) would be associated with more support for integration among minority participants. Results bore out these predictions. Theoretical and policy implications are discussed.


Development of Binomial Sequential Sampling Plans for Sugarcane Aphid (Hemiptera: Aphididae) in Commercial Grain Sorghum.
The sugarcane aphid (Melanaphis sacchari Zehntner) is a significant economic pest of grain sorghum (Sorghum bicolor (L.) Moench) in the Southern United States. Current nominal and research-based economic thresholds are based on estimates of mean aphids per leaf. Because enumerating aphids per leaf is potentially time consuming, binomial sequential sampling plans for M. sacchari were developed that allow users to quickly classify the economic status of field populations and determine when an economic threshold has been exceeded. During 2016 and 2017, counts of M. sacchari were recorded from 281 sampling events in 140 sorghum fields located in six states (Oklahoma, Kansas, Texas, Arkansas, Louisiana, Mississippi) . Regression analysis was used to describe the relationships between the mean M. sacchari density per two-leaf sample and proportion of plants infested with one or more aphids. Tally thresholds of T50 and T100 aphids per two-leaf sample were selected based on goodness of fit and practicality. Stop lines for both tally thresholds were developed for selected economic thresholds using Wald's sequential probability ratio test. Model validations using an additional 48 fields demonstrated that reliable classification decisions could be made with an average of 11 samples regardless of location. This sampling system, when adopted, can allow users to easily and rapidly determine when M. sacchari infestations need to be treated.


Nutrient enrichment and phytoplankton toxicity influence a diversity of complex dynamics in a fear-induced plankton-fish model.
In this paper, we contemplate the dynamics of an aquatic system consisting of three interacting species, phytoplankton, zooplankton, and fish. We assume that the evading risk of fish predation induces fear in zooplankton species, which affects its growth dynamics radically. On the other hand, zooplankton develop an anti-predator defense by taking temporary refuge. Interestingly, the system potentially exhibits multi-stable configurations under identical ecological conditions by allowing different bifurcation scenarios, including multiple saddle-node and transcritical bifurcations with varying levels of nutrients, strength of phytoplankton toxicity, zooplankton refuge size and the cost of fear imposed by fish population. Further, by adding Gaussian white noise, we have extended the deterministic system to its stochastic version. We find that white noise appears to regulate the survival and extinction of model species. Comprehensive numerical simulations are consistent with mathematical results prognosticated by linear analysis. Overall, our study may provide a new insight into the mechanisms of emergence and mitigation of plankton blooms.


Utilization of macrominerals and trace elements in pregnant heifers with distinct feed efficiencies.
The objective of the study was to evaluate utilization of dietary minerals and trace elements in pregnant heifers with distinct residual feed intakes (RFI). Feed intake, body weight (BW), and body composition traits were recorded in 36 crossbred heifers over a period of 37 wk, starting shortly after weaning at 8.3 (0.10; standard deviation) mo of age with an average BW of 276 (7.8) kg. Both BW and body composition were monitored regularly throughout the study, whereas individual feed intake was assessed during the last 84 d of the trial. Data recorded were used to calculate RFI for each heifer. Heifers were ranked based on RFI and assigned to high (n=14) or low (n=10) RFI groups. After the RFI study, 24 selected heifers [age 18.2 (0.14) mo; 87.5 (4.74) d in gestation; 497 (8.5) kg of BW] were used in an indirect digestibility trial (lignin as internal marker). Heifers were fed a ration containing corn silage, haylage, and a mineral premix in which Ca, P, K, Na, Mg, S, Cu, Fe, Mn, Mo, Se, Zn, and Co were provided in the diet according to National Research Council requirements of pregnant replacement heifers. The digestibility trial lasted 1 wk, during which samples of feces were gathered twice daily, and blood and liver biopsy samples were collected on the last day. We noted no significant differences between low- and high-RFI heifers in dry matter digestibility. Apparent absorption of Cu, Zn, and Mn was increased in heifers with low RFI, and apparent absorption of Co tended to be greater for these animals. Concentrations of macrominerals and trace elements in serum of pregnant heifers were similar for both groups except for Se, which was increased in the serum of low-RFI heifers. Liver concentrations of Cu, Fe, Mn, Mo, Se, and Zn did not differ between low- and high-RFI heifers. In conclusion, whereas improved absorption of some trace elements (Cu, Zn, Mn, and Co) and increased Se serum concentration appear to be associated with superior feed efficiency in pregnant heifers, further studies are needed to investigate the causality of such relationships.


The study of developmental capacity of vitrified mouse blastocysts in different straws after transfer to mouse pseudo pregnant.
Vitrification is the commonly used method for long-term storage of pre-implantation mammalian embryos. It is an essential part of assisted reproductive technologies. The re-expansion rate, pregnancy and birth rate of vitrified blastocysts using CPS were compared with OPS and Conventional Straw. Female NMRI mice were injected with Gonadotrophins in order induce them for super ovulation. At that time the mice were sacrified by cervical dislocation and dissected of mouse abdomen. The uterine horns were existed blastocysts were collected in PBS and randomly allocated to four groups: vitrification in CPS, conventional straw, OPS and untreated controls. The vitrification solution was EFS40%. After storage for 1 month in liquid nitrogen, the blastocysts were thawed in 0.5 M sucrose for in vitro culture in M16 medium. After 6 h of culture, the numbers of expanded blastocysts was recorded and ready for transfer to uterus of pseudo pregnant mouse. The re-expansion rate of the CPS group (72.1%) was significantly higher (p < 0.05) than OPS (52.55) and C.S. (38.6%) groups. The pregnancy (70%) and birth rate (45%) of blastocysts in CPS were similar to those of fresh blastocysts (80% and 45.5%) and the pregnancy (10%) and birth rate (5.1%) in Conventional Straws lower than OPS (20 and 7.5%), but were not significantly different. Mouse blastocysts vitrified using CPS had a better result compared with OPS and Conventional Straw. The value of CPS for vitrification of blastocysts may also merit investigation.


Staff attitudes towards older patients with cognitive impairment: need for improvements in acute care.
To explore the attitudes held by staff working in acute care units towards patients aged 70 years or older with cognitive impairment, and to explore factors associated with negative attitudes. Hospital staff attitudes towards older patients with cognitive impairment are of concern as older people are the main hospital users, and because staff attitudes influence care quality and uptake of evidence-based care. A cross-sectional survey design was used to collect data from staff (n = 391). Staff attitudes were not explicitly negative. However, higher perceived strain in caring for older patients with cognitive impairment, higher perceived prevalence of these patients in the ward, being younger and working as an assistant nurse were associated with negative attitudes. A majority of staff reported that these patients received the best possible care, but few reported formally assessing cognitive status or working with evidence-based care protocols. Staff characteristics associated with negative attitudes were described and staff perception that patients received best hospital care, despite limited cognitive assessments and care guidelines, indicate areas for improvement. Supporting young staff and assistant nurses, and implementing cognitive assessments and evidence-based guidelines can promote positive attitudes and best practice.


A review of the cost of cardiovascular disease.
In Canada, 74,255 deaths (33% of all deaths) in 2003 were due to cardio-vascular disease (CVD). As one of the most costly diseases, CVD represents a major economic burden on health care systems. The purpose of the present study was to review the literature on the economic costs of CVD in Canada and other developed countries (United States, Europe and Australia) published from 1998 to 2006, with a focus on Canada. Of 1656 screened titles and abstracts, 34 articles were reviewed including six Canadian studies and 17 American studies. While considerable variation was observed among studies, all studies indicated that the costs of treating CVD-related conditions are significant, outlining a convincing case for CVD prevention programs.


Effect of breast cancer phenotype on diagnostic performance of MRI in the prediction to response to neoadjuvant treatment.
The estimation of response to neoadjuvant chemotherapy (NAC) is useful in the surgical decision in breast cancer. We addressed the diagnostic reliability of conventional MRI, of diffusion weighted imaging (DWI) and of a merged criterion coupling morphological MRI and DWI. Diagnostic performance was analysed separately in different tumor subtypes, including HER2+ (human epidermal growth factor receptor 2)/HR+ (hormone receptor) (hybrid phenotype). Two-hundred and twenty-five patients underwent MRI before and after NAC. The response to treatment was defined according to the RECIST classification and the evaluation of DWI with apparent diffusion coefficient (ADC). The complete pathological response - pCR was assessed (Mandard classification). Tumor phenotypes were Luminal (63.6%), Triple Negative (16.4%), HER2+ (7.6%) or Hybrid (12.4%). After NAC, pCR was observed in 17.3% of cases. Average ADC was statistically higher after NAC (p<0.001) among patients showing pCR vs. those who had not pCR. The RECIST classification showed adequate performance in predicting the pCR in Triple Negative (area under the receiver operating characteristic curve, ROC AUC=0.9) and in the HER2+ subgroup (AUC=0.826). Lower performance was found in the Luminal and Hybrid subgroups (AUC 0.693 and 0.611, respectively), where the ADC criterion yielded an improved performance (AUC=0.787 and 0.722). The coupling of morphological and DWI criteria yielded maximally improved performance in the Luminal and Hybrid subgroups (AUC=0.797 and 0.761). The diagnostic reliability of MRI in predicting the pCR to NAC depends on the tumor phenotype, particularly in the Luminal and Hybrid subgroups. In these cases, the coupling of morphological MRI evaluation and DWI assessment may facilitate the diagnosis.


Educational testing and validity of conclusions in the scholarship of teaching and learning.
Validity and its integral evidence of reliability are fundamentals for educational and psychological measurement, and standards of educational testing. Herein, we describe these standards of educational testing, along with their subtypes including internal consistency, inter-rater reliability, and inter-rater agreement. Next, related issues of measurement error and effect size are discussed. This article concludes with a call for future authors to improve reporting of psychometrics and practical significance with educational testing in the pharmacy education literature. By increasing the scientific rigor of educational research and reporting, the overall quality and meaningfulness of SoTL will be improved.


Workplace Productivity Loss and Indirect Costs Associated With Preterm Birth in the United States.
To estimate workplace productivity loss and indirect costs in the year after birth among individuals who deliver preterm in the United States. This retrospective, observational cohort study estimated workplace productivity loss and indirect costs for individuals aged 18-55 years with an inpatient delivery between January 1, 2016, and September 30, 2021, using data from the Merative MarketScan Commercial Claims and Encounters database and the Health and Productivity Management database. Workdays lost and costs attributable to medical-related absenteeism, workplace absenteeism (defined as sick leave, leave, recreational leave, Family Medical Leave Act); disability (defined as short-term and long-term disability), and aggregate workplace productivity loss, a combined outcome measure, were compared between propensity-score-matched birth cohorts: preterm birth (before 37 weeks of gestation) and full-term birth (at or after 37 weeks of gestation). Outcomes were also compared between the full-term birth cohort and preterm birth subgroups (before 32 weeks of gestation and before 34 weeks of gestation). Estimations of indirect costs assumed an 8-hour workday. Costs were inflated to December 2021 U.S. dollars. In total, 37,522 individuals were eligible for medical-related absenteeism, 1,028 for workplace absenteeism, 7,880 for disability, and 396 for aggregate workplace productivity loss after propensity score matching. Compared with full-term birth, preterm birth was associated with more workdays lost and costs in the year after childbirth attributable to medical-related absenteeism (differences of 4.2 days and $1,045, P <.001) and disability (differences of 2.8 days and $422, P <.001). Preterm birth was not associated with workplace absenteeism (differences of 1.4 days and $347, P =.787) and aggregate workplace productivity loss (differences of 5.2 days [ P =.080] and $1,021 [ P =.093]). Numerical differences were greater in magnitude and inversely related to gestational age at birth across outcomes. Preterm birth was associated with medical-related absenteeism, disability claims, and indirect costs in the year after birth compared with full-term birth.


Persistence of human immunodeficiency virus (HIV) type 1 DNA in peripheral blood despite prolonged suppression of plasma HIV-1 RNA in children.
Human immunodeficiency virus (HIV) type 1 DNA in peripheral blood mononuclear cells (PBMC) was quantified in 31 children who received efavirenz, nelfinavir, and 1 or 2 nucleoside reverse-transcriptase inhibitors for > or =2 years and in whom undetectable plasma HIV-1 RNA levels (< 50 copies/mL) were sustained, to determine the usefulness of HIV-1 DNA as a marker of virus suppression. The median baseline HIV-1 DNA level was 750 copies/10(6) PBMC. After initiation of highly active antiretroviral therapy (HAART), HIV-1 DNA levels decreased gradually, reaching a plateau from week 80 through week 104 (median HIV-1 DNA level, 263 copies/10(6) PBMC). Children who had plasma HIV-1 RNA levels < 50 copies/mL after receiving HAART for 8 weeks (n=16) had persistently lower quantities of intracellular HIV-1 DNA than children whose HIV-1 RNA levels reached < 50 copies/mL after 8 weeks of HAART (n=15). The median half-life for intracellular HIV-1 DNA was 60 weeks. Thus, despite prolonged maintenance of undetectable levels of plasma HIV-1 RNA, HIV-1 DNA remains detectable in PBMC of children and may be a useful marker of further virus suppression.


Effects of dauricine and lidocaine alone or combined on electrophysiological properties of canine Purkinje fibers.
Dauricine (Dau) 1 to 30 mumol/L produced the concentration-dependent depressions in the APA, Vmax, MDP, and prolongations of APD50 and APD90 as well as ERP of the isolated canine cardiac Purkinje fibers (PF). The automaticity and excitation were significantly reduced at concentration of 30 mumol/L. The effects of Dau on all action potential parameters of PF were observed at all stimulation frequencies (60, 75, 100, 150 beats/min). Lidocaine (Lid) markedly shortened APD50 of PF at concentration of 30 mumol/L and also shortened APD90, ERP and significantly depressed APA, Vmax at 100 mumol/L. When perfused in combination with Dau, Lid appreciably shortened APD50 and APD90, and lightly abbreviated ERP prolonged by Dau.


Preliminary evidence for a glucan acceptor in the yeast Candida albicans.
Two membrane preparation containing glucan synthase activity were obtained by lysis of regenerating sphaeroplasts (enzyme A) or mechanical breakage (enzyme B) of yeast (Candida albicans) cells. The reaction products of both enzymes (glucans A and B respectively) were characterized as linear beta-1,3-linked glucans on the basis of chemical and enzymic analysis. In addition, two pools of glucan could be distinguished in glucan A preparations on the basis of their susceptibility to an exoglucanase. In no case were the reaction products synthesized de novo; rather the radioactive chains were added to the non-reducing end of non-radioactive preformed glucan chains or to an acceptor of a different nature. At least some of the performed chains of glucan A, but not those of glucan B, showed a free reducing terminal. Glucan A preparations were endowed with endoglucanase activity, which, under appropriate conditions, released glucose, laminaribiose and laminaritriose. These sugars were also found in cell-wall autolysates. On the basis of the origin of both enzyme preparations it is suggested that glucan molecules are synthesized while they are bound to a non-glucan acceptor that is subsequently excised, presumably by cell-wall-associated glucanases.


An anatomical study of the proximal aspect of the medial femoral condyle to define the proximal-distal condylar length.
Despite its possible role in knee arthroplasty, the proximal-distal condylar length (PDCL) of the femur has never been reported in the literature. We conducted an anatomic study of the proximal aspect of the medial femoral condyle to propose a method for measuring the PDCL. Inspection of dried bone specimens was carried out to assure the most proximal condylar margin (MPCM) as the eligible starting point to measure the PDCL. Simulation surgery was performed on seven pairs of cadaveric knees to verify the clinical application of measuring the PDCL after locating the MPCM. Interobserver reliability of this procedure was also analyzed. Observation of the bone specimens showed that the MPCM is a concavity formed by the junction of the distal end of the supracondylar ridge and the proximal margin of the medial condyle. This anatomically distinctive structure made the MPCM an unambiguous landmark. The cadaveric simulation surgical dissection demonstrated that the MPCM is easily accessed in a surgical setting, making the measurement of the PDCL plausible. The intraclass correlation coefficient was 0.78, indicating good interobserver reliability for this technique. This study has suggested that the PDCL can be measured based on the MPCM in a surgical setting. PDCL measurement might be useful in joint line position management, selection of femoral component sizes, and other applications related to the proximal-distal dimension of the knee. Further investigation is required.


Bacterial Community Associated with Healthy and Diseased Pacific White Shrimp (Litopenaeus vannamei) Larvae and Rearing Water across Different Growth Stages.
Bacterial communities are called another "organ" for aquatic animals and their important influence on the health of host has drawn increasing attention. Thus, it is important to study the relationships between aquatic animals and bacterial communities. Here, bacterial communities associated with Litopenaeus vannamei larvae at different healthy statuses (diseased and healthy) and growth stages (i.e., zoea, mysis, and early postlarvae periods) were examined using 454-pyrosequencing of the 16S rRNA gene. Bacterial communities with significant difference were observed between healthy and diseased rearing water, and several bacterial groups, such as genera Nautella and Kordiimonas could also distinguish healthy and diseased shrimp. Rhodobacteraceae was widely distributed in rearing water at all growth stages but there were several stage-specific groups, indicating that bacterial members in rearing water assembled into distinct communities throughout the larval development. However, Gammaproteobacteria, mainly family Enterobacteriaceae, was the most abundant group (accounting for more than 85%) in shrimp larvae at all growth stages. This study compared bacterial communities associated with healthy and diseased L. vannamei larvae and rearing water, and identified several health- and growth stage-specific bacterial groups, which might be provided as indicators for monitoring the healthy status of shrimp larvae in hatchery.


Infectious Disease Teleconsultation to the Deployed U.S. Military From 2017-2022.
The ADvanced VIrtual Support for OpeRational Forces (ADVISOR) program is a synchronous telemedicine system developed in 2017 to provide 24/7 remote expert support to U.S. Military and NATO clinicians engaged in medical care in austere locations. Infectious disease (ID) remains the highest consulted service since 2018 and is currently staffed by 10 adult and pediatric ID physicians within the Military Health System. We conducted a retrospective review of the ID ADVISOR calls between 2017 and 2022 to identify trends and better prepare military ID physicians to address urgent ID consultations in overseas settings. Health records of the ID consultations between July 2017 and January 2022 were reviewed for local caregiver and patient demographics, case descriptions, consultant recommendations, and outcomes. A "not research" determination was made by the Brooke Army Medical Center Human Research Protections Office. ID physicians received 57 calls for 60 urgent patient consultations. Most calls were from countries in the Middle East or in Southwest Asia (United States Central Command (USCENTCOM)), followed by countries in Africa (United States Africa Command (USAFRICOM)). The majority of patients were active duty U.S. Military and were generally male with median age of 25 years. All consults involved an initial phone consultation and 30% continued over email. Ninety percent of the calls were initiated by physicians, and the median time from injury or illness-onset to consult was 3 days. Seventy percent of the consult questions involved treatment and further diagnostics, but one-third of cases required assistance with management of disease prevention. Multidrug-resistant or nosocomial infections, animal or bite exposure, malaria and malaria prevention, febrile illness, and blood-borne pathogen exposure accounted for 63% of the consults. Collaboration with other specialties took place in a minority of cases, and follow-up contact was recommended 20% of the time. Most recommendations involved adjusting drug regimens or further testing. Medical evacuation was only recommended in five of the cases. Although there was limited ability for follow-up, no known deaths occurred. A high proportion of calls to the ID ADVISOR line are relevant to the overlapping content areas of infection prevention, force protection, and outbreak response. Most patients requiring urgent ID consultation were managed successfully without evacuation. The current military-unique ID fellowship curriculum is consistent with the encountered diagnoses per the ID ADVISOR line, and high-yield individual topics have been identified.


A mouse kindling model of perimenstrual catamenial epilepsy.
Catamenial epilepsy is caused by fluctuations in progesterone-derived GABA(A) receptor-modulating anticonvulsant neurosteroids, such as allopregnanolone, that play a significant role in the pathophysiology of epilepsy. However, there is no specific mouse model of catamenial epilepsy. In this study, we developed and characterized a mouse model of catamenial epilepsy by using the neurosteroid-withdrawal paradigm. It is hypothesized that seizure susceptibility decreases when neurosteroid levels are high (midluteal phase) and increases during their withdrawal (perimenstrual periods) in close association with GABA(A) receptor plasticity. A chronic seizure condition was created by using the hippocampus kindling model in female mice. Elevated neurosteroid levels were induced by sequential gonadotropin treatment, and withdrawal was induced by the neurosteroid synthesis inhibitor finasteride. Elevated neurosteroid exposure reduced seizure expression in fully kindled mice. Fully kindled mice subjected to neurosteroid withdrawal showed increased generalized seizure frequency and intensity and enhanced seizure susceptibility. They also showed reduced benzodiazepine sensitivity and enhanced neurosteroid potency, similar to the clinical catamenial seizure phenotype. The increased susceptibility to seizures and alterations in antiseizure drug responses are associated with increased abundance of the α4 and δ subunits of GABA(A) receptors in the hippocampus. These findings demonstrate that endogenous neurosteroids protect against seizure susceptibility and their withdrawal, such as that which occurs during menstruation, leads to exacerbation of seizure activity. This is possibly caused by specific changes in GABA(A) receptor-subunit plasticity and function, therefore providing a novel mouse model of human perimenstrual catamenial epilepsy that can be used for the investigation of disease mechanisms and new therapeutic approaches.


Microfilaments and microtubules in calcium ionophore-induced secretion of lysosomal enzymes from human polymorphonuclear leukocytes.
Human peripheral blood leukocytes (PMN) are induced to release lysosomal enzymes by the calcium ionophore A23187 in the presence but not the absence of extracellular Ca++. Whereas secretion induced by particulate or immune stimuli is accompanied by an increase in visible microtubules and is inhibitable by colchicine, secretion induced by A23187 and Ca++ was not accompanied by an increase in microtubule numbers and was not inhibited by colchicine. Ca++ did not appear to regulate microtubule assembly in these cells since resting PMN had a mean of 22.3 +/- 2.0 microtubules in the centriolar region as compared to 22.3 +/- 1.1 in ionophore-treated cells and 24.9 +/- 1.5 in cells exposed to ionophore and 1 mM Ca++. Bipolar filaments, 10 nm thick and 300--400 nm long, were numerous in the pericortical cytoplasm of cells exposed to both reagents. Microtubules in these cells were decorated with an electron-opaque fibrillar material. PMN exposed to A23187 and Ca++ were contracted in two directions at right angles to each other: (a) Contractions parallel to the plasma membrane resulted in extensive plication of the cell membrane. The cytoplasm subjacent to the plicae contained dense filamentous webs. Plication was prevented by cytochalasin B or reversed by subsequent exposure to an endocytic stimulus such as zymosan. (b) Contractions perpendicular to the plasma membrane, toward the cytocenter, resulted in the formation of vacuoles in normal PMN and of membrane invaginations in cytochalasin B-treated PMN. Whereas contractions parallel to the plasma membrane could occur in the absence of enzyme release (ionophore alone) and enzyme release could occur in the absence of such contractions (ionophore plus calcium plus cytochalasin B), contraction toward the cytocenter occurred in all experimental conditions in which significant enzyme release was obtained. Thus, lysosomal enzyme secretion in PMN involves contractile movements in the plasma membrane toward the lysosomes rather than the reverse. These calcium-mediated contractile events are mediated by cytochalasin B-insensitive microfilaments but not by microtubule assembly.


Kicking Triturus arntzeni when it's down: large-scale nuclear genetic data confirm that newts from the type locality are genetically admixed.
We collected nuclear DNA data (52 markers) with next-generation sequencing for nine Triturus newt specimens, including the holotype and two of the paratypes of T. arntzeni, from the type locality at Vrtovać in eastern Serbia. We compare these data to a reference set composed of the four crested newt species distributed in eastern Serbia namely T. cristatus, T. dobrogicus, T. ivanbureschi and T. macedonicus to determine to which of these species the newts from the type locality of T. arntzeni should be attributed. The majority of alleles in individuals from Vrtovać is derived from T. macedonicus, but a considerable number of T. ivanbureschi alleles is also present; alleles typical for T. cristatus and T. dobrogicus are found at low frequency. Accordingly, we interpret Vrtovać as a T. macedonicus - T. ivanbureschi hybrid population, albeit not composed of F1 hybrids but of genetically admixed individuals derived through multiple generations of backcrossing. The data support the notion that the name T. arntzeni should not be applied to a species newly distinguished in T. karelinii sensu lato (to which the name T. ivanbureschi has been given). We conclude that because of the hybrid nature of the individuals from Vrtovać, the name T. arntzeni should be placed not only in the synonymy of T. macedonicus but also in the synonymy of T. ivanbureschi. In this study we demonstrate that next-generation sequencing can provide high quality data for type material with degraded DNA and therefore can play an important role in taxonomy.


The scientific advisory system: some observations.
The scientific advisory system is effective on limited technical questions, and such questions provide much of its work. On broad technical questions, however, the scientific advisory system is not effective. Unfortunately this category includes most of the crucial environmental questions. Finally, the advisory system, as presently constituted, combined with the multiple functions of the scientific establishment, is detrimental in important ways to the process of technical decision-making in this country. This is because the combined effect of the advisory system and the establishment is to impede the development of a more effective and comprehensice role for the scientific community in the technical decision-making process.


Targeting the phosphatidylinositol 3-kinase/AKT pathway for the treatment of multiple myeloma.
Multiple myeloma is the second most hematological malignancy, accounting for more than 10% of all blood cancers and 2% of annual cancer-related deaths due to lack of curable drugs. Novel and molecularly targeted anti-MM drugs are in urgent need. The phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathway plays a critical regulatory role in multiple myeloma pathophysiology, including survival, proliferation, migration, angiogenesis, as well as drug resistance, and has emerged as a key therapeutic target. Many potent inhibitors targeting this pathway have been developed and some have been moved for clinical evaluations for multiple myeloma. In this review, we highlighted the role of the PI3K/AKT pathway in the pathogenesis of multiple myeloma, and current advances in drug discovery for this class of inhibitors. Discovery strategies toward the PI3K/AKT inhibitors were also discussed.