Prenatal diagnosis of VACTERL association: a case report.
A prenatal diagnosis of VACTERL association, a combination of vertebral (V), anal (A), cardiac (C), tracheoesophageal (TE), renal (R) and limb (L) anomalies was made at 30 weeks of gestation, based on the sonographic demonstration of vertebral defects, bilateral renal agenesis, and left lower limb defects. Additionally, severe oligohydramnios and fetal growth restriction were also documented. After proper counseling, elective termination of pregnancy was done, resulting in a stillborn infant with multiple malformations compatible with the VACTERL association. The postnatal X-ray and autopsy revealed verterbral defects, anorectal atresia with undetermined sex, cardiac defect of ventricular septal defect, tracheal agenesis with distal atresia of esophagus, bilateral renal agenesis, and limbs defects. The chromosomal study revealed normal, 46,XY. This report emphasizes the important role of prenatal ultrasound in the diagnosis and management of this disorder.


Measurement of breast density with dual X-ray absorptiometry: feasibility.
Dual x-ray absorptiometry (DXA) was used to quantify breast density with a phantom and with cadaveric breasts. With DXA, percentage of fat correlated with percentage of glandular density of the phantom (r > 0.998) and with density at mammography (r(adjusted) = 0.83). DXA precision (SD) was 0.5% without and 1.1% with breast repositioning. DXA devices can be used to accurately and precisely estimate breast tissue density.


A formal medication reconciliation programme in a haemodialysis unit can identify medication discrepancies and potentially prevent adverse drug events.
Patients on haemodialysis have been identified as high-risk for medication discrepancies and adverse drug events. Medication reconciliation is an important patient safety initiative to prevent adverse drug events. The primary objective of our study was to determine the number and types of medication discrepancies and drug therapy problems (DTPs) identified in patients on haemodialysis. Our second objective was to assess the potential clinical impact and severity of all unintentional medication discrepancies identified. Patients in an academic haemodialysis unit were interviewed to obtain a best possible medication history (BPMH) between May and August 2010. The BPMH was documented and discrepancies were identified, classified and resolved with the interprofessional team. An interprofessional panel conducted a discrepancy clinical impact assessment for potential adverse drug events. Two hundred and twenty-eight patients on haemodialysis were interviewed and 512 discrepancies were identified for 151 patients (3.4 discrepancies per patient). Of these, 174 (34%) were undocumented intentional discrepancies and 338 (66%) were unintentional discrepancies. The unintentional discrepancies were classified as 21% omissions, 36% commissions and 43% incorrect dose/frequency. Most drug therapy problems were related to patient taking a medication that was not indicated (25%), medication required but patient not taking (25%), patient not willing to take the medication as prescribed (28%) or incorrect dosing of a drug (20%). Overall, 6% of discrepancies were classified as clinically significant potential adverse drug events. Medication discrepancies appear to be common in patients on haemodialysis. Formal interprofessional medication reconciliation practice models are essential to identify discrepancies and prevent patients from experiencing adverse drug events.


Social deprivation as a risk factor for late presentation of proliferative diabetic retinopathy.
The aim of this study was to determine whether social deprivation is a risk factor for late presentation of patients with proliferative diabetic retinopathy and whether it affects their access to urgent laser treatment. Using a 2:1 case: control design, 102 patients referred to a UK teaching hospital as part of the UK Diabetic Retinopathy National Screening Programme were identified for the period between 1 June 2010 to 1 June 2013. Social deprivation was scored using the Index of Multiple Deprivation 2010. Additional variables considered included age, duration of disease, ethnicity, and HbA1c at time of referral. The cases comprised 34 patients referred with proliferative (grade R3) retinopathy with a control group of 68 patients with lower retinopathy grades; two control patients were excluded due to incomplete data. On univariate analysis, R3 retinopathy was associated with higher social deprivation (P<0.001, Mann-Whitney U-test), and with higher HbA1c (11.5% vs 8.4%; P<0.001, Mann-Whitney U-test). Forward stepwise multivariable analysis showed that the association of R3 retinopathy with deprivation was significant even after adjusting for HbA1c (P=0.016). On univariate analysis South Asian ethnicity was also identified as being a risk factor for presentation with R3 retinopathy, but this was no longer significant when HbA1c was adjusted for in a forward stepwise logistic regression analysis. In our cohort social deprivation appears to be associated with late presentation of proliferative diabetic retinopathy. Our study supports the need to target these groups to reduce preventable blindness and to identify strategies which overcome barriers to care.


[Carbohydrate linkage with the peptide portion of the ristomycin A molecule].
The mannose monosaccharide residue in ristomycin A is bound via C-3 phenolic oxygen of the 1,2,3,5-substituted ring with actinoidinic amino acid. The tetrassacharide branch is attached to the only phenolic oxygen of triaminotricarboxylic acid. The ristosaminyl residue is linked via one of the two aliphatic hydroxyl groups of the same amino acid. By the location of neutral carbohydrates risotmycin A does not differ from ristocetin A.


A Rare Case of Hemoglobin Bart's Hydrops Fetalis due to Uniparental Disomy of Chromosome 16.
Hemoglobin (Hb) Bart's hydrops fetalis is the most severe form of α-thalassemia and is usually inherited in an autosomal recessive manner. We report a case of Hb Bart's hydrops fetalis due to uniparental disomy of chromosome 16. Antenatal screening showed a low maternal mean corpuscular volume (MCV), while paternal MCV was normal. The fetus was found to have a thickened nuchal translucency during first trimester screening for Down's syndrome. Mid-trimester fetal anomaly ultrasound scan showed fetal cardiomegaly with pericardial effusion, scalp edema, ascites and an elevated middle cerebral arterial peak systolic velocity (MCA PSV). Multiplex polymerase chain reaction (PCR) on DNA from amniocentesis showed that the fetus was homozygous for South East Asian (SEA) type 2 α-globin gene deletion. Chromosome microarray (CMA) showed two regions of absence of heterozygosity (AOH) on the terminal p and q arm of chromosome 16. The rare occurrence of Hb Bart's hydrops fetalis caused by maternal uniparental disomy should be considered in cases of fetal hydrops even in cases where paternal MCV is normal.


KIT immunohistochemistry and mutation status in gastrointestinal stromal tumours (GISTs) evaluated for treatment with imatinib.
With the availability of effective but expensive treatment in the form of imatinib, accurate diagnosis of gastrointestinal stromal tumour (GIST) is extremely important. The aims of this study were: to describe the clinicopathological, immunohistochemical and molecular features of cases referred to a cancer centre with a possible diagnosis of GIST; to identify pitfalls in the performance and interpretation of KIT immunohistochemistry; to define the role of KIT mutation testing in making a diagnosis of GIST. Morphological review, KIT immunohistochemistry and mutation testing were performed on all cases referred with a diagnosis of GIST or where the diagnosis was under serious consideration on the basis of KIT immunopositivity with a view to treating with imatinib. Thirty-seven cases met the inclusion criteria. Of these, 26 were classified as GIST and 11 as non-GIST. Most GISTs showed strong diffuse membranous, cytoplasmic or paranuclear KIT immunopositivity. Some non-GISTs demonstrated patchy cytoplasmic KIT immunopositivity related to the immunohistochemical protocol used in the external laboratory, which led to erroneous diagnoses of GIST in nine (24%) cases. KIT mutations involving exons 11 or 9 were identified in 22 (88%) GISTs tested and none of the non-GISTs. An accurate diagnosis of GIST can be made on clinicopathological and immunohistochemical criteria without the need for mutational analysis in most cases, provided proper attention is paid to the immunohistochemical protocol used and, most importantly, control material. False-positive diagnoses of GIST potentially leading to inappropriate treatment with imatinib are more common than missed diagnoses.


Electrical stimulation effects on tenderness of five muscles from Hampshire x Rambouillet crossbred lambs with the callipyge phenotype.
The objective of this study was to determine effects of electrical stimulation (ES) on muscle quality and sensory traits of 12 Hampshire x Rambouillet callipyge lambs. One side of each carcass was randomly assigned to an ES treatment of 550 V and 60 Hz of electricity for 2 s on and 2 s off 15 times. The other side was a nonstimulated control (NES). Heated calpastatin, sarcomere length, myofibrillar fragmentation index (MFI), Warner-Bratzler shear (WBS), and trained sensory panel values were measured on the semitendinosus (ST), semimembranosus (SM), longissimus (ML), supraspinatus (SP), and triceps brachii (TB) muscles. Electrically stimulating the carcass sides induced a more rapid (P = .001) pH decline in the longissimus muscle, and ES sides had a brighter (P = .001) red color of loineye than nonstimulated sides. At d 14 of storage (2 degrees C), the TB had the highest (P < .05) MFI value, indicating more protein degradation, and the ST and ML muscles had the lowest MFI (P = .008). Regardless of ES treatment, SM and ML had the highest (P < .05) WBS values. The ST muscle had higher (P < .05) WBS values than the SP but did not differ (P > .05) from the TB muscle. Electrical stimulation had no effect on WBS or any trained sensory panel values (P > .05). The percentage of loin chops rated slightly tender or better was improved 30 to 34% by electrical stimulation (P < .05). The ML muscle was scored lower (P < .05) in sustained juiciness compared with the SM, SP, and TB but did not differ (P > .05) from the ST muscle. The SM and ML muscles were rated lower (P < .05) in initial and sustained tenderness scores than other muscles. Tenderness scores were higher (P < .05) for the TB than for the SP but did not differ (P > .05) from the ST muscle. Electrically stimulating callipyge carcasses improves the tenderness of loin chops by increasing the percentage of chops rated from slightly tough to slightly tender.


Predictive value of cord blood hematological indices and hemoglobin Barts for the detection of heterozygous alpha-thalassemia-2 in an African-Caribbean population.
Cord blood hemoglobin Barts (HbBarts) and hemocytometric indices may be used for classification of newborns into those without alpha-thalassemia-2 (alphaalpha/alphaalpha) and with heterozygous alpha-thalassemia-2 (-alpha(3.7)/alphaalpha). We investigated by logistic regression analysis whether the combination of HbBarts and hemocytometric indices improves classification compared with classification based on a single analyte. HbBarts percentages and hemocytometric indices were determined in cord blood of 208 consecutive newborns in Curaçao (Netherlands Antilles). Of these, 157 had alphaalpha/alphaalpha and 51 had -alpha(3.7)/alphaalpha, as established by DNA analysis. Between-group differences were significant for erythrocytes, mean cell volume, mean cell hemoglobin (MCH), mean cell hemoglobin concentration, platelets, hemoglobin F(0) (HbF(0)), and HbBarts. The Logit equation of the logistic regression model, using MCH (pg) and HbBarts (%), was: 42.7164 + 5.7916(HbBarts) - 1.3110(MCH). A sensitivity of 100% was reached at a Logit value of -3.70. The corresponding specificity was 62.2%, and the predictive value of a positive test (PV+) was 46.3% (95% confidence interval, 37.0-55.7%). The relative information gains were as follows: 88% for the HbBarts-MCH combination, 26% for MCH (not significant), and 0% for HbBarts compared with the 24.6% -alpha(3.7)/alphaalpha prevalence. Combined use of cord blood HbBarts and MCH improves classification compared with classification based on single hemocytometric indices.


PAMDB: a comprehensive Pseudomonas aeruginosa metabolome database.
The Pseudomonas aeruginosaMetabolome Database (PAMDB, http://pseudomonas.umaryland.edu) is a searchable, richly annotated metabolite database specific to P. aeruginosa. P. aeruginosa is a soil organism and significant opportunistic pathogen that adapts to its environment through a versatile energy metabolism network. Furthermore, P. aeruginosa is a model organism for the study of biofilm formation, quorum sensing, and bioremediation processes, each of which are dependent on unique pathways and metabolites. The PAMDB is modelled on the Escherichia coli (ECMDB), yeast (YMDB) and human (HMDB) metabolome databases and contains >4370 metabolites and 938 pathways with links to over 1260 genes and proteins. The database information was compiled from electronic databases, journal articles and mass spectrometry (MS) metabolomic data obtained in our laboratories. For each metabolite entered, we provide detailed compound descriptions, names and synonyms, structural and physiochemical information, nuclear magnetic resonance (NMR) and MS spectra, enzymes and pathway information, as well as gene and protein sequences. The database allows extensive searching via chemical names, structure and molecular weight, together with gene, protein and pathway relationships. The PAMBD and its future iterations will provide a valuable resource to biologists, natural product chemists and clinicians in identifying active compounds, potential biomarkers and clinical diagnostics.


Clinical correlation recommended: accuracy of clinician versus radiologic interpretation of the imaging of orbital lesions.
Purpose: To assess the accuracy of radiographic interpretation between the clinician and radiologist when compared to histopathology of orbital lesions. Methods: A retrospective chart review of patients at the University of California Davis Eye Center who underwent orbitotomy from 1/1/2000 to 5/22/2019 was performed. Charts with a preoperative imaging report, preoperative clinical assessment including the clinician's interpretation of imaging, and histopathologic diagnosis were included. The specific diagnoses were grouped into related classes of pathology for the analysis. The clinical and radiologic assessments were compared against the final histopathologic diagnosis for concordance. A concordance analysis was performed. Results: 242 patients (mean age 49 years, 53.5% female) were reviewed. Of these records 185 documented the clinician's clinical impression, the radiology report, as well as the histopathology report. The clinician's preoperative assessment had substantial agreement [kappa = 0.72 (0.65,0.79)] with the final histopathologic result and was correct in 75.7% (140/185) of cases whereas the radiology report was correct in 52.4% (97/185) with a moderate level of agreement [kappa = 0.47 (0.39, 0.55)]. In 49.2% (91/185) of cases the final histopathology correlated with both the clinical impression and radiology report [kappa = 0.58 (0.55, 0.61)]. Conclusions: The accurate interpretation of orbital imaging is a challenge and histopathologic examination remains the gold standard for diagnosis. While orbital imaging is a valuable diagnostic tool the interpretation of these studies is most accurate when conducted in the context of the patient's medical history, clinical exam, and with the physician most familiar with various orbital lesions.


Super-resolution imaging of the natural killer cell immunological synapse on a glass-supported planar lipid bilayer.
The glass-supported planar lipid bilayer system has been utilized in a variety of disciplines. One of the most useful applications of this technique has been in the study of immunological synapse formation, due to the ability of the glass-supported planar lipid bilayers to mimic the surface of a target cell while forming a horizontal interface. The recent advances in super-resolution imaging have further allowed scientists to better view the fine details of synapse structure. In this study, one of these advanced techniques, stimulated emission depletion (STED), is utilized to study the structure of natural killer (NK) cell synapses on the supported lipid bilayer. Provided herein is an easy-to-follow protocol detailing: how to prepare raw synthetic phospholipids for use in synthesizing glass-supported bilayers; how to determine how densely protein of a given concentration occupies the bilayer's attachment sites; how to construct a supported lipid bilayer containing antibodies against NK cell activating receptor CD16; and finally, how to image human NK cells on this bilayer using STED super-resolution microscopy, with a focus on distribution of perforin positive lytic granules and filamentous actin at NK synapses. Thus, combining the glass-supported planar lipid bilayer system with STED technique, we demonstrate the feasibility and application of this combined technique, as well as intracellular structures at NK immunological synapse with super-resolution.


Going full circle: phase-transition thermodynamics of ionic liquids.
We present the full enthalpic phase transition cycle for ionic liquids (ILs) as examples of non-classical salts. The cycle was closed for the lattice, solvation, dissociation, and vaporization enthalpies of 30 different ILs, relying on as much experimental data as was available. High-quality dissociation enthalpies were calculated at the G3 MP2 level. From the cycle, we could establish, for the first time, the lattice and solvation enthalpies of ILs with imidazolium ions. For vaporization, lattice, and dissociation enthalpies, we also developed new prediction methods in the course of our investigations. Here, as only single-ion values need to be calculated and the tedious optimization of an ion pair can be circumvented, the computational time is short. For the vaporization enthalpy, a very simple approach was found, using a surface term and the calculated enthalpic correction to the total gas-phase energy. For the lattice enthalpy, the most important constituent proved to be the calculated conductor-like screening model (COSMO) solvation enthalpy in the ideal electric conductor. A similar model was developed for the dissociation enthalpy. According to our assessment, the typical error of the lattice enthalpy would be 9.4 kJ mol(-1), which is less than half the deviation we get when using the (optimized) Kapustinskii equation or the recent volume-based thermodynamics (VBT) theory. In contrast, the non-optimized VBT formula gives lattice enthalpies 20 to 140 kJ mol(-1) lower than the ones we assessed in the cycle, because of the insufficient description of dispersive interactions. Our findings show that quantum-chemical calculations can greatly improve the VBT approaches, which were parameterized for simple, inorganic salts with ideally point-shaped charges. In conclusion, we suggest the term "augmented VBT", or "aVBT", to describe this kind of theoretical approach.


Feasibility of using the Vero SBRT system for intracranial SRS.
The Vero SBRT system was benchmarked in a planning study against the Novalis SRS system for quality of delivered dose distributions to intracranial lesions and assessing the Vero system's capacity for SRS. A total of 27 patients with one brain lesion treated on the Novalis system, with 3 mm leaf width MLC and C-arm gantry, were replanned for Vero, with a 5 mm leaf width MLC mounted on an O-ring gantry allowing rotations around both the horizontal and vertical axis. The Novalis dynamic conformal arc (DCA) planning included vertex arcs, using 90° couch rotation. These vertex arcs cannot be reproduced with Vero due to the mechanical limitations of the O-ring gantry. Alternative class solutions were investigated for the Vero. Additionally, to distinguish between the effect of MLC leaf width and different beam arrangements on dose distributions, the Vero class solutions were also applied for Novalis. In addition, the added value of noncoplanar IMRT was investigated in this study. Quality of the achieved dose distributions was expressed in the conformity index (CI) and gradient index (GI), and compared using a paired Student's t-test with statistical significance for p-values ≤ 0.05. For lesions larger than 5 cm3, no statistical significant difference in conformity was observed between Vero and Novalis, but for smaller lesions, the dose distributions showed a significantly better conformity for the Novalis (ΔCI = 13.74%, p = 0.0002) mainly due to the smaller MLC leaf width. Using IMRT on Vero reduces this conformity difference to nonsignificant levels. The cutoff for achieving a GI around 3, characterizing a sharp dose falloff outside the target volume was 4 cm3 for Novalis and 7 cm3 for Vero using DCA technique. Using noncoplanar IMRT, this threshold was reduced to 3 cm3 for the Vero system. The smaller MLC and the presence of the vertex fields allow the Novalis system to better conform the dose around the lesion and to obtain steeper dose falloff outside the lesion. Comparable dosimetric characteristics can be achieved with Vero for lesions larger than 3 cm3 and using IMRT.


Isolation and identification of a novel microcystin-degrading bacterium from a biological sand filter.
A novel bacterium capable of degrading two microcystin analogues, microcystin-LR and -LA (MCLR and MCLA), was isolated from a biological sand filter which was previously shown to effectively remove these toxins from source waters. Based on phylogenetic analysis of the 16S rRNA gene sequence, the isolated organism, LH21, most likely belonged to the genus Sphingopyxis and of the previously cultured species clustered with Sphingopyxis witflariensis. Using polymerase chain reaction (PCR), isolate LH21 was shown to contain homologues to each of the four genes, mlrA, mlrB, mlrC and mlrD previously associated with the degradation of MCLR by Sphingomonas sp. ACM-3962. Isolate LH21 was able to effectively degrade MCLR and MCLA in batch experiments under environmentally relevant conditions, with complete removal observed within 5h after re-exposure of the toxins.


Kyanite-Andalusite Equilibrium from 700{degrees} to 800{degrees}C.
The metastable kyaniteandalusite equilibrium in the Al(2)SiO(5) system has been reversed at 700 degrees , 750 degrees , and 800 degrees C at elevated water pressures, with a variety of natural and synthetic kyanites and andalusites as starting materials. Sillimanite, the stable form of Al(2)SiO(5) under these conditions, did not appear. The value of the transition pressure at 750 degrees C is 6.6 +/- 0.4 kilobars, several kilobars below pressures given by several convergent previous determinations. The Al(2)SiO(5) pressure-temperature triple point now indicated lies far from the points found by others. The revised aluminum silicate phase diagram indicates that many rocks crystallized at lower pressures than formerly thought possible.


Gelatin microgel-coated balloon catheter with enhanced delivery of everolimus for long-term vascular patency.
In-stent restenosis (ISR) after percutaneous coronary intervention is a major reason for limited long-term patency due to complex neointimal proliferation caused by vascular injury. Drug-coated balloon (DCB) has been developed to treat various cardiovascular diseases including ISR by providing anti-proliferative drugs into blood vessel tissues. However, a significant proportion of the drug is lost during balloon tracking, resulting in ineffective drug delivery to the target region. In this study, we report an everolimus-coated balloon (ECB) using everolimus-loaded gelatin-hydroxyphenyl propionic acid microgel (GM) with enhanced everolimus delivery to vascular walls for long-term patency. GM with high drug loading (> 97%) was simply prepared by homogenizing enzyme-mediated crosslinked hydrogels. The optimal condition to prepare GM-coated ECB (GM-ECB) was established by changing homogenization time and ethanol solvent concentration (30 ∼ 80%). In vitro sustained everolimus release for 30 d, and cellular efficacy using smooth muscle cells and vascular endothelial cells were evaluated. Additionally, an in vivo drug transfer levels of GM-ECB using rabbit femoral arteries were assessed with reduced drug loss and efficient drug delivery capability. Finally, using ISR-induced porcine models, effective in vivo vascular patency 4 weeks after treatment of ECBs was also confirmed. Thus, this study strongly demonstrates that GM can be used as a potential drug delivery platform for DCB application. STATEMENT OF SIGNIFICANCE: We report an ECB using everolimus-loaded GM prepared by homogenization of enzymatic cross-linked hydrogel. GM showed efficient drug loading (> 97 %) and controllable size. GM-ECB exhibited potential to deliver everolimus in a sustained manner to target area with drug efficacy and viability against SMC and EC. Although GM-ECB had much lower drug content compared to controls, animal study demonstrated enhanced drug transfer and reduced drug loss of GM-ECB due to the protection of encapsulated drugs by GM, and the possible interaction between GM and endothelium. Finally, vascular patency and safety were assessed using ISR-induced porcine models. We suggest an advanced DCB strategy to alleviate rapid drug clearance by bloodstream while improving drug delivery for a long-term vascular patency.


Novel Finnish-enriched variants causing severe hypercholesterolemia and their clinical impact on coronary artery disease.
Severe hypercholesterolemia (LDL-cholesterol ≥ 5 mmol/l) is a major risk factor for coronary artery disease (CAD). The etiology incudes both genetic and nongenetic factors, but persons carrying mutations in known hypercholesterolemia-associated genes are at significantly higher CAD risk than non-carriers. Yet, a significant proportion of mutation carriers remains undetected while the assessment of genetic candidate variants in clinical practice is challenging. To address these challenges, we set out to test the utility of a practical approach to leverage data from a large reference cohort, the FinnGen Study encompassing 356,082 persons with extensive longitudinal health record information, to aid the clinical evaluation of single genetic candidate genes variants detected by exome sequence analysis in a target population of 351 persons with severe hypercholesterolemia. We identified 23 rare missense mutations in known hypercholesterolemia genes, 3 of which were previously described mutations (LDLR Pro309Lysfs, LDLR Arg595Gln and APOB Arg3527Gln). Subsequent in silico and clinical assessment of the remaining 20 variants pinpointed two likely hypercholesterolemia-associated variants in LDLR (Arg574Leu and Glu626Lys) and one in LDLRAP1 (Arg151Trp). Heterozygous carriers of the novel LDLR and LDLRAP1 variants received statin treatment more often than non-carriers (OR 2.1, p = 1.8e-6 and OR 1.4, p = 0.001) and untreated carriers had higher risk for ischemic heart disease (OR 2.0, p = 0.03 and OR 1.8, p = 0.008). Our data elucidate the wide spectrum of genetic variants impacting hypercholesterolemia and demonstrate the utility of a large reference population to assess the heterogeneous impact of candidate gene variants on cardiovascular disease risk.


Thermoregulation in endothermic insects.
On the basis of body weight, most flying insects have higher rates of metabolism, and hence heat production, than other animals. However, rapid rates of cooling because of small body size in most cases precludes appreciable endothermy. The body temperature of small flies in flight is probably close to ambient temperature, and that of flying butterflies and locusts is 5 degrees to 10 degrees C above ambient temperature. Many moths and bumblebees are insulated with scales and hair, and their metabolism during flight can cause the temperature of the flight muscles to increase 20 degrees to 30 degrees C above ambient temperature. Curiously, those insects which (because of size, insulation) retain the most heat in the thorax during flight, also require the highest muscle temperature in order to maintain sufficient power output to continue flight. The minimum muscle temperature for flight varies widely between different species, while the maximum temperature varies over the relatively narrow range of 40 degrees to 45 degrees C. As a consequence, those insects that necessarily generate high muscle temperatures during flight must maintain their thoracic temperature within a relatively narrow range during flight. Active heat loss from the thorax to the abdomen prevents overheating of the flight motor and allows some large moths to be active over a wide range of ambient temperatures. Bumblebees similarly transfer heat from the flight musculature into the abdomen while incubating their brood by abdominal contact. Many of the larger insects would remain grounded if they did not actively increase the temperature of their flight muscles prior to flight. Male tettigoniid grasshoppers elevate their thoracic temperature prior to singing. In addition, some of the social Hymenoptera activate the "flight" muscles specifically to produce heat not only prior to flight but also during nest temperature regulation. During this "shivering" the "flight" muscles are often activated in patterns different from those during flight. The muscles contract primarily against each other rather than on the wings. However, the rate of heat production during shivering and flight is primarily a function of the action potential frequency rather than of the patterns of activation. Thermoregulation is a key factor in the energetics of foraging of some of the flower-visiting insects. The higher their muscle temperature the more flowers they can visit per unit time. When food supplies are ample, bees may invest relatively large amounts of energy for thermoregulation. While shivering to maintain high body temperatures during the short intervals they are perched on flowers (as well as while in the nest), bumblebees often expend energy at rates similar to the rates of energy expenditure in flight. Unlike vertebrates, which usually regulate their body temperature at specific set points, the body temperature of insects is labile. It often appears to be maintained near the lower temperature at which the muscles are able to perform the function at hand. The insects' thermal adaptations may not differ as much from those of vertebrates as previously supposed when size, anatomy, and energy requirements are taken into account.


Using same-hospital readmission rates to estimate all-hospital readmission rates.
Since October of 2012, Medicare's Hospital Readmissions Reduction Program has fined 2,200 hospitals a total of $500 million. Although the program penalizes readmission to any hospital, many institutions can only track readmissions to their own hospitals. We sought to determine the extent to which same-hospital readmission rates can be used to estimate all-hospital readmission rates after major surgery. We evaluated 3,940 hospitals treating 741,656 Medicare fee-for-service beneficiaries undergoing CABG, hip fracture repair, or colectomy between 2006 and 2008. We used hierarchical logistic regression to calculate risk- and reliability-adjusted rates of 30-day readmission to the same hospital and to any hospital. We next evaluated the correlation between same-hospital and all-hospital rates. To analyze the impact on hospital profiling, we compared rankings based on same-hospital rates with those based on all-hospital rates. The mean risk- and reliability-adjusted all-hospital readmission rate was 13.2% (SD 1.5%) and mean same-hospital readmission rate was 8.4% (SD 1.1%). Depending on the operation, between 57% (colectomy) and 63% (CABG) of hospitals were reclassified when profiling was based on same-hospital readmission rates instead of on all-hospital readmission rates. This was particularly pronounced in the middle 3 quintiles, where 66% to 73% of hospitals were reclassified. In evaluating hospital profiling under Medicare's Hospital Readmissions Reduction Program, same-hospital rates provide unstable estimates of all-hospital readmission rates. To better anticipate penalties, hospitals require novel approaches for accurately tracking the totality of their postoperative readmissions.


Curcumin I protects the dopaminergic cell line SH-SY5Y from 6-hydroxydopamine-induced neurotoxicity through attenuation of p53-mediated apoptosis.
Oxidative stress (OS) plays a pivotal role in the pathogenesis of Parkinson's disease (PD). 6-Hydroxydopamine (6-OHDA) is a neurotoxin used to induce oxidative cell death of dopaminergic neurons in experimental models of PD. Curcumin I, or diferuloylmethane is a pure compound isolated from Curcuma longa Linn. that has been reported to have neuroprotective properties. The precise mechanism, however, remains unclear. This study aims to elucidate the mechanisms by which curcumin I exerts its effects, using 6-OHDA-induced neurotoxicity in the human dopaminergic cell line SH-SY5Y. In our experiments, pretreatment with curcumin I improved cell viability, and significantly reduced reactive oxygen species (ROS). Further investigations revealed a reduction of p53 phosphorylation and decrease of the Bax/Bcl-2 ratio, as measured by mRNA expression and protein level. Taken together, these findings indicate that curcumin I protects dopaminergic neurons from 6-OHDA-induced toxicity via the reduction of ROS production, and subsequent attenuation of p53 phosphorylation and reduction of the Bax/Bcl-2 ratio.


The alveolar fibroproliferative response in moderate to severe COVID-19-related acute respiratory distress syndrome and one-year follow up.
COVID-19-related acute respiratory distress syndrome (ARDS) can lead to long-term pulmonary fibrotic lesions. Alveolar fibroproliferative response (FPR) is a key factor in the development of pulmonary fibrosis. N-terminal-peptide of procollagen III (NT-PCP-III) is a validated biomarker for activated FPR in ARDS. This study aimed to assess the association between dynamic changes in alveolar FPR and long-term outcomes, as well as mortality in COVID-19-ARDS patients. We conducted a prospective cohort study of 154 COVID-19-ARDS patients. We collected bronchoalveolar lavage (BAL) and blood samples for measurement of 17 pulmonary fibrosis biomarkers, including NT-PCP-III. We assessed pulmonary function and chest computed tomography (CT) at 3 and 12 months after hospital discharge. We performed joint modelling to assess the association between longitudinal changes in biomarker levels and mortality at day 90 after starting mechanical ventilation. 154 patients with 284 BAL samples were analysed. Of all patients, 40% survived to day 90, of whom 54 completed the follow-up procedure. A longitudinal increase in NT-PCP-III was associated with increased mortality (HR 2.89, 95% CI: 2.55-3.28; p<0.001). Forced vital capacity and diffusion for carbon monoxide were impaired at 3 months but improved significantly at one year after hospital discharge (p=0.03 and p=0.004, respectively). There was no strong evidence linking alveolar FPR during hospitalization and signs of pulmonary fibrosis in pulmonary function or chest CT images during one-year follow-up. In COVID-19-ARDS patients, alveolar FPR during hospitalization was associated with higher mortality but not with the presence of long-term fibrotic lung sequelae within survivors.


[On the treatment of acute arterial circulatory disturbances of the retina and optic nerve (author's transl)].
After discussing basic therapeutic principles for the treatment of acute arterial occlusions of the retina and N. opticus (fibrinolysis, anticoagulation, corticoids, vasoactive drugs) the problems in employment of vasodilating drugs and the necessity of concomitant administration of strophanthine or digitalisglycosides are outlined. Results obtained after the administration of 3,7-Dimethyl-1-(5-oxo-hexyl)-xanthine (Pentoxifylline, Trental) in the treatment of 49 cases suffering from occlusion of the retinal vessels and 23 cases with vascular occlusion in the N. opticus are reported and commented with a view to the bad prognosis of these serious disorders, 55% of the patients showing definite improvement of visual acuity. Therapeutic guidelines resulting from these experiences are presented.


Whole-cell recording of calcium release-activated calcium (CRAC) currents in human T lymphocytes.
In T lymphocytes, depletion of Ca(2+) from the intracellular Ca(2+) store leads to activation of plasmalemmal Ca(2+) channels, called Calcium Release-Activated Calcium (CRAC) channels. CRAC channels play important role in regulation of T cell proliferation and gene expression. Abnormal CRAC channel function in T cells has been linked to severe combined immunodeficiency and autoimmune diseases. Studying CRAC channel function in human T cells may uncover new molecular mechanisms regulating normal immune responses and unravel the causes of related human diseases. Electrophysiological recordings of membrane currents provide the most accurate assessment of functional channel properties and their regulation. Electrophysiological assessment of CRAC channel currents in Jurkat T cells, a human leukemia T cell line, was first performed more than 20 years ago, however, CRAC current measurements in normal human T cells remains a challenging task. The difficulties in recording CRAC channel currents in normal T cells are compounded by the fact that blood-derived T lymphocytes are much smaller in size than Jurkat T cells and, therefore, the endogenous whole-cell CRAC currents are very low in amplitude. Here, we give a step-by-step procedure that we routinely use to record the Ca(2+) or Na(+) currents via CRAC channels in resting human T cells isolated from the peripheral blood of healthy volunteers. The method described here was adopted from the procedures used for recording the CRAC currents in Jurkat T cells and activated human T cells.


The cyclin-dependent kinase inhibitor (CDKI) flavopiridol disrupts phorbol 12-myristate 13-acetate-induced differentiation and CDKI expression while enhancing apoptosis in human myeloid leukemia cells.
Interactions between the cyclin-dependent kinase inhibitor (CDKI) flavopiridol (FP) and phorbol 12-myristate 13-acetate (PMA) were examined in U937 human leukemia cells in relation to differentiation and apoptosis. Simultaneous, but not sequential, exposure of U937 cells to 100 nM FP and 10 nM PMA significantly increased apoptosis manifested by characteristic morphological features, mitochondrial dysfunction, caspase activation, and poly(ADP-ribose) polymerase cleavage while markedly inhibiting cellular differentiation, as reflected by diminished plastic adherence and CD11b expression. Enhanced apoptosis in U937 cells was associated with an early caspase-independent increase in cytochrome c release and accompanied by a substantial decline in leukemic cell clonogenicity. Moreover, PMA/FP cotreatment significantly increased apoptosis in HL-60 promyelocytic leukemia cells and in U937 cells ectopically expressing the Bcl-2 protein. In U937 cells, coadministration of FP blocked PMA-induced expression and reporter activity of the CDKI p21WAF/CIP1 and triggered caspase-mediated cleavage of the CDKI p27KIP1. Coexposure to FP also resulted in a more pronounced and sustained activation of the mitogen-activated protein kinase kinase/extracellular signal-regulated protein kinase cascade after PMA treatment, although disruption of this pathway by the mitogen-activated protein kinase kinase 1 inhibitor U0126 did not prevent potentiation of apoptosis. FP accelerated PMA-mediated dephosphorylation of the retinoblastoma protein (pRb), an event followed by pRb cleavage culminating in the complete loss of underphosphorylated pRb (approximately Mr 110,000) by 24 h. Finally, gel shift analysis revealed that coadministration of FP with PMA for 8 h led to diminished E2F/pRb binding compared to the effects of PMA alone. Collectively, these findings indicate that FP modulates the expression/activity of multiple signaling and cell cycle regulatory proteins in PMA-treated leukemia cells and that such alterations are associated with mitochondrial damage and apoptosis rather than maturation. These observations also raise the possibility that combining CDKIs and differentiation-inducing agents may represent a novel antileukemic strategy.


Dynamics and Viability of Airborne Respiratory Syncytial Virus under Various Indoor Air Conditions.
The factors governing the viability of airborne viruses embedded within respiratory particles are not well understood. This study aimed to investigate the relative humidity (RH)-dependent viability of airborne respiratory syncytial virus (RSV) in simulated respiratory particles suspended in various indoor air conditions. We tested airborne RSV viability in three static indoor air conditions, including sub-hysteresis (RH < 39%), hysteresis (39% < RH < 65%), and super-hysteresis (RH > 65%) air as well as in three dynamic indoor air conditions, including the transitions between the static conditions. The dynamic conditions were hysteresis → super-hysteresis → hysteresis, sub-hysteresis → hysteresis, and super-hysteresis → hysteresis. We found that after 45 min of particle aging in static conditions, the viability of RSV in sub-hysteresis, hysteresis, and super-hysteresis air was 0.72% ± 0.06%, 0.03% ± 0.006%, and 0.27% ± 0.008%, respectively. After 45 min of aging in dynamic conditions, the RSV viability decreased for particles that remained in a liquid (deliquesced) state during aging when compared with particles in a solid (effloresced) state. The decreased viability of airborne RSV for deliquesced particles is consistent with prolonged exposure to elevated aqueous solutes. These results represent the first measurements of the survival of airborne RSV over particle aging time, with equal viability in low, intermediate, and high RHs at 5 and 15 min and a V-shaped curve after 45 min.


Association between Mycoplasma-specific polymerase chain reaction assay results and oral bacterial contamination of bronchoalveolar lavage fluid samples from dogs with respiratory tract disease: 121 cases (2005-2012).
To determine whether an association exists between oral bacterial contamination of bronchoalveolar lavage fluid (BALF) and positive PCR assay results for the detection of Mycoplasma spp in BALF samples of dogs with lower respiratory tract (LRT; portion from the trachea to the lungs) disease. Retrospective case series. 121 dogs with LRT disease. Procedures-Medical records from January 2005 to April 2012 were reviewed. Dogs with LRT disease that had BALF samples evaluated by use of Mycoplasma-specific PCR assay, bacterial culture, and cytologic examination were included. Information on signalment, final diagnoses, and BALF testing results was extracted. 83 (68.6%) dogs had BALF samples with negative PCR assay results for Mycoplasma spp, and 38 (31.4%) had positive results. The BALF samples with cytologic evidence of oral bacterial contamination were 5.1 times as likely to have positive Mycoplasma-specific PCR assay results as were noncontaminated samples. Compared with hound or herding dogs, other breeds were 13.6 times as likely to have positive PCR assay results. Dogs with bronchitis were less likely than dogs with other LRT diseases to have positive Mycoplasma-specific PCR assay results. No significant association was found between Mycoplasma-specific PCR assay results and bacterial culture results. In dogs with LRT disease, Mycoplasma-specific PCR assay results for BALF samples should be interpreted in terms of possible oral bacterial contamination. Mycoplasma-specific PCR assay of BALF samples from herding dogs, hound dogs, and dogs with bronchitis may be less rewarding than for other dogs with LRT disease.


[Inhibitory effects of ursolic acid on diabetic retinopathy in mice].
To explore the protective effect and its mechanisms of intravitreal injection of ursolic acid (UA) on retinal injury in diabetic retinopathy (DR) mice. DR model was set up by using alloxan. 60 C57BL/6 male mice (clean animal, aged 12 weeks) were randomly and evenly divided into 6 groups: blank control group, DR model group, positive control group (triamcinolone acetonide), UA intervention groups (low, moderate and high dose). Mice were killed one week after drug intervention and the eyeballs were removed to prepare paraffin section. Retinal new blood vessel was examined by histopathological methods. The mRNA and protein expressions of vascular endothelial growth factor (VEGF), cyclooxygenase 2 (COX-2), and Matrix Metalloproteinase 2 (MMP-2) in retinal tissues were detected by real-time PCR and Western blotting, respectively. The superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were detected by related kits. The HE and PAS staining showed uniform inner limiting membrane structure and orderly vascular endothelial cells in blank control group, and neovascularization was seldom detected. In DR model group, numerous new blood vessels were formed beyond inner limiting membrane. UA intervention group with low dose had almost the same results with DR group, while the results of UA intervention group with high dose were similar with positive control group, in which the inner limiting membrane structure was uniform and new blood vessels were seldom seen. The mRNA expressions of VEGF, COX-2, and MMP-2 in DR model group were significantly higher than those of blank control group (P<0.001) and UA intervention group with moderate and high dose (P<0.001). The protein expressions of VEGF, COX-2 and MMP-2 in DR model group were significantly higher than those of blank control group (P<0.001) and UA intervention group with low, moderate and high dose (all P<0.05). The MDA content of retinal tissue in DR model group was significantly higher than those of blank control group (P=0.001) and UA intervention groups with moderate and high dose (P=0.026, 0.007). The activity of SOD was significantly lower in DR model group than those of blank control group (P<0.001) and UA intervention groups with low, moderate and high dose (P=0.040, 0.021, 0.003). UA may inhibit the formation of new blood vessels through reducing the expressions of VEGF, COX-2 and MMP-2 in retinal tissues, and promote a remission from DR by obvious resistance to oxidative stress.


A role of interferon-gamma (IFN-gamma) in tumor immunity: T cells with the capacity to reject tumor cells are generated but fail to migrate to tumor sites in IFN-gamma-deficient mice.
IFN-gamma-deficient (IFN-gamma-/-) mice induce potent in vitro immune responses such as anti-allo mixed lymphocyte reaction and CTL responses, whereas they often fail to exhibit in vivo immunity. Here, we investigated whether there exists a defect in tumor rejection responses and if so, which process of responses is impaired. IFN-gamma-/- and wild-type (WT) BALB/c mice were immunized with attenuated syngeneic CSA1M tumor cells. The capacity of T cells to mediate tumor protection was examined in Winn assays to assess the growth of tumor cells admixed with tumor-sensitized T cells. Splenic T cells from both groups of mice exhibited comparable levels of tumor-neutralizing activity. When portions of immunized mice were directly challenged with viable tumor cells, tumor rejection was induced only in WT mice. CD4(+) and CD8(+) T-cell infiltration were observed at the site of tumor challenge in WT mice, whereas such a T-cell infiltration did not occur in IFN-gamma-/- mice. Similarly, splenic T cells from interleukin 12-treated CSA1M-bearing IFN-gamma-/- and WT mice neutralized tumor cells at comparable efficacies in Winn assays. However, the migration of these T cells to tumor masses and the resultant interleukin 12-induced tumor regression took place in WT mice, but neither intratumoral T-cell infiltration nor tumor regression occurred in IFN-gamma-/- mice. These results indicate a critical requirement for IFN-gamma in the process of inducing T-cell migration to tumor sites rather than of generating antitumor protective T cells.


[Primary tumors of the mesentery and greater omentum].
Diseases of mesentery and greater omentum are commonly the result of lymph node metastases from an abdominal carcinoma, or lymph node involvement as part of a general lymphatic malignancy. Primary tumours are rare. Cysts occur predominantly in children. In addition, one may find mesenchymal benign or malignant tumours, which produce clinical symptoms at a later age. Plain abdominal films, barium meals and angiograms fail to show evidence of a space-occupying lesion, or produce only indirect signs. Only ultrasound or CT can demonstrate these tumours directly.


Collection and analysis of a Parkinson speech dataset with multiple types of sound recordings.
There has been an increased interest in speech pattern analysis applications of Parkinsonism for building predictive telediagnosis and telemonitoring models. For this purpose, we have collected a wide variety of voice samples, including sustained vowels, words, and sentences compiled from a set of speaking exercises for people with Parkinson's disease. There are two main issues in learning from such a dataset that consists of multiple speech recordings per subject: 1) How predictive these various types, e.g., sustained vowels versus words, of voice samples are in Parkinson's disease (PD) diagnosis? 2) How well the central tendency and dispersion metrics serve as representatives of all sample recordings of a subject? In this paper, investigating our Parkinson dataset using well-known machine learning tools, as reported in the literature, sustained vowels are found to carry more PD-discriminative information. We have also found that rather than using each voice recording of each subject as an independent data sample, representing the samples of a subject with central tendency and dispersion metrics improves generalization of the predictive model.


Oncoprotein expression in human synovial tissue: an immunohistochemical study of different types of arthritis.
Based on the fact that synovial lining cells have some properties of transformed-appearing cells, we have examined the expression of Myc, Myb, Fos, Jun and Ras oncoproteins in synovial tissues from patients with different types of arthritis. Formalin-fixed and paraffin-embedded sections of synovial tissue from 12 patients with rheumatoid arthritis (RA), 14 with reactive arthritis (ReA), nine with other seronegative arthritis (OSA), seven with bacterial arthritis (BA), eight with probable bacterial arthritis (PBA) and eight with osteoarthritis (OA) were studied using the immunoperoxidase staining technique. The oncoproteins studied were expressed both in the synovial lining layer and in the sublining layer, consisting of lymphocytes, other inflammatory cells and blood vessels. Among the six disease entities, RA and OA appeared to be the most distinct, whereas the results obtained for ReA and OSA, and on the other hand for BA and PBA, closely resembled each other. The expression of Myc, Myb, Fos and Jun was significantly correlated both to the degree of synovial hypercellularity and the synovial lymphocytic infiltration. For Ras, such a correlation could not be seen. We conclude that we find no evidence of a cell lineage-specific or a disease-specific abnormality of proto-oncogene products in RA, and the expression of these oncoproteins is consistent with inflammation rather than with any primary abnormality of cell growth.


Oral ciprofloxacin therapy for gram-negative bacillary osteomyelitis.
Gram-negative osteomyelitis frequently responds poorly to conventional therapy. Ciprofloxacin displays excellent in vitro activity against gram-negative bacilli and offers the potential for outpatient therapy. In this ongoing study, ciprofloxacin therapy is being evaluated for the treatment of gram-negative osteomyelitis. Twenty-three patients (16 men and seven women) have been treated under the protocol (750 mg orally twice daily for 1.5 to six months), and 14 patients have completed therapy. All patients had either growth on bone cultures from an open or percutaneous biopsy, or an arthrocentesis to confirm the diagnosis. Involved sites included ankle or tibia (seven patients), vertebra (four patients), hip (five patients), metatarsal (four patients), phalanx (two patients), and metacarpal (one patient); 16 patients had chronic disease, and seven patients had acute disease. Patients had a total of 28 gram-negative bacilli, 12 gram-positive cocci, and one anaerobic gram-negative rod, for an average of 1.8 pathogens per patient. Eighteen of the 28 gram-negative bacilli were Pseudomonas species. The geometric mean minimal inhibitory concentration for all the gram-negative bacilli was 0.15 microgram/ml. The geometric mean minimal inhibitory concentration for the gram-positive isolates was 0.41 microgram/ml. All patients who completed therapy experienced a cure, with a mean follow-up of 6.1 months. Infections in all patients, except for two who are still taking ciprofloxacin, are resolving, both clinically and radiologically. One patient who was not eligible for the protocol experienced a superinfection with methicillin-resistant Staphylococcus aureus. Side effects have included urticaria, lethargy, nausea, and transient elevations of liver and renal function test results. Overall, ciprofloxacin therapy was well tolerated. This study suggests that ciprofloxacin holds promise for the outpatient treatment of gram-negative osteomyelitis.


DROD: bio-robotic drill/sampler for planetary subterranean exploration: experiments and challenges.
Key features for space exploration equipment, and in particular drills and sampling mechanisms, are low weight, small size, and energy efficiency. These characteristics are substantially required not only in reducing the spaceship flight cost, but also in extending the exploration time on the extraterrestrial bodies. This article experimentally investigates the feasibility of a novel drill bioinspired by wood-wasp and sand-fish lizard as an integrated robotic solution for rover exploration tasks. A new penetration depth of 820 mm in term of reciprocation drilling technique has been achieved by the proposed dual reciprocation and oscillation drill (DROD), especially with the new enhancements such as miniature sample compartment and toothed stems. Additionally, a first sampling experiment with DROD has been performed and a sample amount of 20 grams and size of 30 cm3 has been collected successfully. Finally, the article provides developments for integration of DROD with rovers for future exploration missions and potentials for horizontal drilling for subterranean applications.&#xD.


Acupuncture for stress urinary incontinence in adults.
The use of acupuncture for stress urinary incontinence is increasing in frequency, especially in Asian area. However, its effectiveness and side effects have not been evaluated. To assess the effectiveness and side effects of acupuncture for stress urinary incontinence in adults. We searched the Cochrane Incontinence Group Specialised Register (searched 28 January 2013), EMBASE, AMED, Chinese Biomedical Literature Database (CBM), Chinese Acupuncture Trials Register and China National Knowledge Infrastructure (CNKI) (all searched 20 February 2013). In addition, we searched the reference lists of relevant articles and contacted authors and trialists in the field. Randomised and quasi-randomised controlled trials of acupuncture interventions without other treatments for the management of stress urinary incontinence for adults. Two review authors independently assessed eligibility, trial quality and extracted data. We meta-analysed data where appropriate. We identified 17 possibly eligible studies but only one small trial with 60 women met our inclusion criteria. The trial compared acupuncture versus midodrine, a drug for treating hypotension. The risk of bias was high as there was no concealment of randomised allocation, and there was no blinding of assessment of outcome. In addition, it was not possible to blind participants or health providers to the interventions. The statistical methods were not described.More women improved in the acupuncture group (73% with acupuncture versus 33% with midodrine; risk ratio (RR) 2.20, 95% confidence interval (CI) 1.27 to 3.81) but the cure rates were low and not statistically significantly different (13% versus 7%; RR 2.00, 95% CI 0.40 to 10.11). There were adverse events in the drug group only. The effect of acupuncture for stress urinary incontinence for adults is uncertain. There is not enough evidence to determine whether acupuncture is more effective than drug treatment.


BRP-170 and BRP190 isoforms of Bruchpilot protein differentially contribute to the frequency of synapses and synaptic circadian plasticity in the visual system of Drosophila.
In the first optic neuropil (lamina) of the optic lobe of Drosophila melanogaster, two classes of synapses, tetrad and feedback, show daily rhythms in the number and size of presynaptic profiles examined at the level of transmission electron microscopy (TEM). Number of tetrad presynaptic profiles increases twice a day, once in the morning and again in the evening, and their presynaptic ribbons are largest in the evening. In contrast, feedback synapses peak at night. The frequency of synapses is correlated with size of the presynaptic element measured as the platform size of so-called T-bars, with T-bar platforms being largest with increasing synapse frequency. The large scaffold protein Bruchpilot (BRP) is a major essential constituent of T-bars, with two major isoforms of 190 and 170 kD forming T-bars of the peripheral neuromuscular junctions (NMJ) synapses and in the brain. In addition to the analysis of cyclic plasticity of tetrad and feedback synapses in wild-type flies, we used TEM to examine daily changes in the size and distribution of synapses within isoform-specific BRP mutants, expressing BRP-190 (BRPΔ170) or BRP-170 (BRPΔ190) only. We found that the number and circadian plasticity of synapses depends on both isoforms. In the BRPΔ190 lacking BRP-190 there was almost 50% less tetrad synapses demonstrable than when both isoforms were present. The lack of BRP-170 and BRP-190 increased and decreased, respectively the number of feedback synapses, indicating that BRP-190 forms most of the feedback synapses. In both mutants, the daily plasticity of tetrad and feedback presynaptic profiles was abolished, except for feedback synapses in BRPΔ190. The oscillations in the number and size of presynaptic elements seem to depend on a different contribution of BRP isoforms in a presynaptic element at different time during the day and night and at various synapse types. The participation of both BRP isoforms may vary in different classes of synapses.


[Use of cannabis among adolescents and young adults in Norway].
Even occasional use of cannabis may increase the risk of serious mental disease (e.g. schizophrenia). Little has been known about the use of cannabis in Norway; especially among adults. A representative sample of young men and women were followed up from they were in their teens in 1992 until they were young adults in 2005. During this 13-year period they were asked four times (through a questionnaire) about use of cannabis, their education, job status, income, marital status, parenthood and parental education and social class. In the mid teens (15 - 16 years), less than 10 % had used cannabis. However, a rather high proportion had their debut in their late teens, as well as throughout their twenties. At the end of the twenties, 40 % of the men and 29 % of the women had used cannabis at some point, and 18 % of the men and 8 % of the women had used cannabis during the previous 12 months. Among the users, 70 % had used the substance 1 - 10 times the last 12 months, while 30 % had a more frequent pattern of use. During the teenage years, there were no associations with parental social class. However, during the twenties, the recruitment base of cannabis use increasingly became socially marginal men who lived aloe and had little education. Use of cannabis is currently rather prevalent in groups of young adults in Norway, in particular among men. The future incidence of psychosis and schizophrenia may be affected by this.


Experimental study on the effect of ammonia on the phase behavior of tetrahydrofuran clathrates.
Clathrate hydrates, ice-like crystalline compounds in which small guest molecules are enclosed inside cages formed by tetrahedrally hydrogen-bonded water molecules, are naturally abundant on Earth and are generally expected to exist on icy celestial bodies. A prototypical example is Saturn's moon Titan, where dissociation of methane clathrates, a major crustal component, could contribute significantly to the replenishment of atmospheric methane. Ammonia is an important clathrate inhibiting agent that may be present (potentially at high concentrations) in Titan's interior. In this study, low-temperature Raman experiments are conducted to examine the dissociation point of tetrahydrofuran clathrates, an ambient-pressure analogue of methane clathrates, over a wide range of ammonia concentrations from 0 to 25 wt %. A phase diagram for the H2O-THF-NH3 system is generated, showing two main results: (i) ammonia lowers the dissociation point of clathrate hydrates to a similar extent compared to the melting of water ice and (ii) THF clathrate exhibits a "liquidus-like" behavior in the presence of ammonia, with a eutectic temperature of about 203.6 K. As temperatures higher than this estimated eutectic are anticipated within Titan's icy crust, these results imply that partial dissociation of clathrates can occur readily and may contribute to outgassing from the interior.


Specific alteration of the Bax/Bcl2 ratio and cytochrome c without execution of apoptosis in the hippocampus of aged baboons.
During ageing, there may be an age-associated loss of particular nerve cells in specific brain areas. Recent studies highlight the role of apoptosis in the normal ageing of the brain, heart, and skeletal muscle. Particular attention is paid to the role of cytochrome c release from mitochondria and alterations in the pro- and anti-apoptotic proteins, Bax and Bcl-2, respectively. The aim of this study was to investigate the potential occurrence of apoptosis in the hippocampus of aged baboons. we have used the baboon as a potential non-human primate model for age-related pathology which afflicts the human brain. The TUNEL staining method was used to characterise the apoptotic cell death. For immunocytochemistry, antibodies directed against activated caspase-3, cytochrome c, Bcl-2 and Bax proteins were used. Our results show that in hippocampi of aged baboons the immunoreactivities of the antiapoptotic agent Bcl-2 was not prominently changed, of the proapoptotic agent Bax was upregulated, of the cytochrome c was redistributed, and that caspase-3 was not activated. The TUNEL-staining method revealed no apoptotic cell death in hippocampi of aged baboons. This demonstrates that a specific alteration of the Bax/Bcl2 ratio occurs in the ageing baboon hippocampus which may directly influence the release of cytochrome c even without commitment to apoptosis.


Red-Emitting Fluorophores as Local Water-Sensing Probes.
Fluorescent probes are known for their ability to sense changes in their direct environment. We introduce here the idea that common red-emitting fluorophores recommended for biological labeling and typically used for simple visualization of biomolecules can also act as reporters of the water content in their first solvent sphere by a simple measurement of their fluorescence lifetime. Using fluorescence spectroscopy, we investigated the excited-state dynamics of seven commercially available fluorophores emitting between 650 and 800 nm that are efficiently quenched by H2O. The amount of H2O in their direct surrounding was modulated in homogeneous H2O-D2O mixtures or, in heterogeneous systems, by confining them into reverse micelles, by encapsulating them into host-guest complexes with cyclodextrins, or by attaching them to peptides and proteins. We found that their fluorescence properties can be rationalized in terms of the amount of H2O in their direct surroundings, which provides a general mechanism for protein-induced fluorescence enhancements of red-emitting dyes and opens perspectives for directly counting water molecules in key biological environments or in polymers.


Structure-Activity Relationships for Rates of Aromatic Amine Oxidation by Manganese Dioxide.
New energetic compounds are designed to minimize their potential environmental impacts, which includes their transformation and the fate and effects of their transformation products. The nitro groups of energetic compounds are readily reduced to amines, and the resulting aromatic amines are subject to oxidation and coupling reactions. Manganese dioxide (MnO2) is a common environmental oxidant and model system for kinetic studies of aromatic amine oxidation. In this study, a training set of new and previously reported kinetic data for the oxidation of model and energetic-derived aromatic amines was assembled and subjected to correlation analysis against descriptor variables that ranged from general purpose [Hammett σ constants (σ(-)), pKas of the amines, and energies of the highest occupied molecular orbital (EHOMO)] to specific for the likely rate-limiting step [one-electron oxidation potentials (Eox)]. The selection of calculated descriptors (pKa, EHOMO, and Eox) was based on validation with experimental data. All of the correlations gave satisfactory quantitative structure-activity relationships (QSARs), but they improved with the specificity of the descriptor. The scope of correlation analysis was extended beyond MnO2 to include literature data on aromatic amine oxidation by other environmentally relevant oxidants (ozone, chlorine dioxide, and phosphate and carbonate radicals) by correlating relative rate constants (normalized to 4-chloroaniline) to EHOMO (calculated with a modest level of theory).


Does proximal location of culprit lesion confer worse prognosis in patients undergoing primary percutaneous coronary intervention for ST elevation myocardial infarction?
ST segment elevation myocardial infarction (STEMI) from proximally located culprit lesion is associated with greater myocardium at jeopardy. In STEMI patients treated with thrombolytics, proximal culprit lesions are known to have worse prognosis. This relation has not been studied in patients undergoing primary percutaneous coronary intervention (PCI). In 3,535 STEMI patients with native coronary artery occlusion pooled from the primary angioplasty in myocardial infarction database, we compared in-hospital and 1-year outcomes between those with proximal (n = 1,606) versus non-proximal (n = 1,929) culprit lesions. Patients with proximal culprits were more likely to die and suffer major adverse cardiovascular events (MACE) during the index hospital stay (3.8% vs 2.2%, P = 0.006; 8.2% vs 5.8%, P = 0.0066, respectively) as well as during 1-year follow-up (6.9% vs 4.5%, P = 0.0013; 22% vs 17%, P = 0.003, respectively) compared to those with non-proximal culprits. After adjustment for baseline differences, proximal culprit was independently predictive of in-hospital death (adjusted odds ratio% 1.58, 95% confidence intervals, CI 1.05-2.40) and MACE (OR 1.41, CI 1.06-1.86), but not 1-year death or MACE. In addition, proximal culprit was independently associated with higher incidence of ventricular arrhythmias and sustained hypotension during the index hospitalization. The univariate impact of proximal culprit lesion on in-hospital death and MACE was comparable to other adverse angiographic characteristics, such as multivessel disease and poor initial thrombolysis in myocardial infarction flow, and greater than that of anterior wall STEMI. In conclusion, proximal location of the culprit lesion is a strong independent predictor of worse in-hospital outcomes in patients with STEMI undergoing primary PCI.


Glatiramer acetate: a review of its use in relapsing-remitting multiple sclerosis and in delaying the onset of clinically definite multiple sclerosis.
Glatiramer acetate is a synthetic analogue of the multiple sclerosis (MS)-associated antigen, myelin basic protein. It is indicated in the EU, US and many other countries to reduce the frequency of relapses in patients with relapsing-remitting MS (RRMS), and for the treatment of patients who have experienced a well defined first clinical episode and are at high risk of developing clinically definite MS or have features of MS on MRI. The efficacy of glatiramer acetate in patients with RRMS has been shown in two randomized, double-blind, multicentre phase III trials. In one trial, glatiramer acetate was associated with a significantly lower mean relapse rate than placebo after 24 months' treatment (primary endpoint). In an ongoing open-label extension to this trial, glatiramer acetate was associated with a sustained reduction in the relapse rate at the 6-, 8- and 15-year follow-ups. In the other trial, the mean cumulative number of gadolinium-enhancing lesions on T1-weighted MRI images was significantly lower in glatiramer acetate versus placebo recipients after 9 months' treatment (primary endpoint). Glatiramer acetate also had generally similar efficacy to subcutaneous interferon (IFN)-beta-1a or IFNbeta-1b in two large randomized, open-label, multicentre phase III trials conducted over 96 weeks or >or=2 years. These data were supported by those from a smaller randomized, open-label phase IV trial that utilized a unique imaging protocol to evaluate the efficacy of glatiramer acetate versus that of IFNbeta-1b over >or=2 years. In these trials, there was no significant difference between glatiramer acetate and IFNbeta recipients in any of the clinical endpoints at study end (e.g. time to first relapse [primary endpoint of the REGARD trial] or risk of relapse [primary endpoint of the BEYOND trial]). Moreover, there was no significant difference between glatiramer acetate and IFNbeta-1b recipients in the median number of combined active lesions per patient per monthly MRI scan during the first 12 months of treatment (primary endpoint of the BECOME trial). In general, there was no significant between-group difference in the majority of other MRI-assessed endpoints in any of the trials. The efficacy of glatiramer acetate in patients with clinically isolated syndrome (CIS) was established in a randomized, double-blind, double-dummy, multicentre phase III trial (the PreCISe trial). In this study, glatiramer acetate was associated with a significantly longer time to conversion to clinically definite MS than placebo (primary endpoint). Glatiramer acetate was generally well tolerated in clinical trials, with most adverse events being mild to moderate in severity. Injection site-related reactions and immediate post-injection systemic reactions were the most frequently observed adverse events associated with glatiramer acetate in clinical studies. In conclusion, glatiramer acetate is a valuable first-line option in the treatment of RRMS, as well as being an option in the treatment of CIS.


A case of cysticercosis cerebri. Neurotologic implications.
Cysticercosis is an unusual disease entity in the United States. A case is reviewed in which a young Mexican-American female presented to an otologist's office with a headache and vertigo. Her symptoms were a manifestation of a single larval cysticercus cyst which had become lodged in the fourth ventricle resulting in an obstructive hydrocephalus and a fatal course. The epidemiology of this disease traces the life cycle of the pork tapeworm from man, normally the definitive host, to the pig, the intermediate host. It is noteworthy that the disease cysticercosis in humans is a deviation from the parasite's normal life cycle in which man serves as the intermediate host. The literature is reviewed. The broad clinical spectrum of this disease's presentation can extend from a single headache to multiple nerve palsies and convulsions. The importance of radiographic evaluation complement fixation testing and CSF evaluation in making a diagnosis is stressed. There is no known medical treatment for this disease. Surgical procedures such as primary excision and shunts to relieve intracranial pressure have had mixed results. The prognosis depends on the location and the number of infesting larva. Special note is made of this disease's endemic status in Mexico and other Latin American countries and the role that this may play in medicine practiced in the southwest United States considering the ever-increasing immigration from this area.


Age and Not the Preferred Limb Influences the Kinematic Structure of Pointing Movements.
In goal-directed movements, effective open-loop control reduces the need for feedback-based corrective submovements. The purpose of this study was to determine the influence of hand preference and aging on submovements during single- and two-joint pointing movements. A total of 12 young and 12 older right-handed participants performed pointing movements that involved either elbow extension or a combination of elbow extension and horizontal shoulder flexion with their right and left arms to a target. Kinematics were used to separate the movements into their primary and secondary submovements. The older adults exhibited slower movements, used secondary submovements more often, and produced relatively shorter primary submovements. However, there were no interlimb differences for either age group or for the single- and two-joint movements. These findings indicate that open-loop control is similar between arms but compromised in older compared to younger adults.


Cancer Antigen 125 (CA125, MUC16) Protein Expression in the Diagnosis and Progression of Pancreatic Ductal Adenocarcinoma.
Pancreatic ductal adenocarcinoma (PDAC) is an aggressive carcinoma, with most patients diagnosed at an advanced stage, with a 5-year survival rate of around 5%. An urgent need exists for identifying better diagnostic, prognostic, and therapeutic markers for this lethal disease. Recently, CA125 has been identified in PDAC, and the aim of this research is to study the changes in CA125 expression during the progression of benign pancreatic tissue (BPT) to PDAC and to assess its value as a biomarker of tumor growth. To address these questions, the cellular levels of CA125 in BPT and PDAC were measured using immunohistochemistry and compared on the basis of tumor staging, and the tissue microarray technology were constructed using resected pancreatic tissues. The staining reactions for each case were evaluated semiquantitatively using the histologic score system. Our investigation demonstrates a consistent and significant upregulation of CA125 during the transition from BPT to PDAC. We also found a direct correlation between CA125 immunohistochemistry score and tumor stage (P=0.02). In conclusion, our data indicate that CA125 plays a direct role in pancreatic carcinogenesis and suggests that it may eventually be used as a diagnostic and/or prognostic biomarker of pancreatic cancer. Prospective studies are recommended to evaluate further the diagnostic and prognostic capabilities of CA125 in PDAC, and further studies are warranted to assess the use of CA125 as a therapeutic marker.


Organisation and chemical neuroanatomy of the African elephant (Loxodonta africana) olfactory bulb.
The olfactory system of mammals can be divided into a main and accessory olfactory system with initial processing for each system occurring in the olfactory bulb. The main and accessory olfactory bulbs have similar structural features, even though they appear to be functionally independent. In mammals the main olfactory bulb (MOB) is also one of two established sites of lifelong generation of new cells. The present study describes the histological and immunohistochemical neuroanatomy of the olfactory bulb of the African elephant (Loxodonta africana). The morphology of MOB of the elephant does not differ significantly from that described in other mammals; however, it lacks the internal plexiform layer. In addition, the glomeruli of the glomerular layer are organised in 2-4 "honey-combed" layers, a feature not commonly observed. The cell types and structures revealed with immunohistochemical stains (parvalbumin, calbindin, calretinin, tyrosine hydroxylase, orexin-A, glial fibrillary acidic protein) were similar to other mammals. Neurogenesis was examined using the neurogenic marker doublecortin. Migration of newly generated cells was observed in most layers of the MOB. No accessory olfactory bulb (AOB) was observed. Based on the general anatomy and the immunohistochemical observations, it is evident that the morphology of the African elephant MOB is, for the most part, similar to that of all mammals, although very large in absolute size.


Asymmetry-induced effects in coupled phase-oscillator ensembles: Routes to synchronization.
A system of two coupled ensembles of phase oscillators can follow different routes to interensemble synchronization. Following a short report of our preliminary results [Phys. Rev. E 78, 025201(R) (2008)], we present a more detailed study of the effects of coupling, noise, and phase asymmetries in coupled phase-oscillator ensembles. We identify five distinct synchronization regions and routes to synchronization that are characteristic of the coupling asymmetry. We show that noise asymmetry induces effects similar to that of coupling asymmetry when the latter is absent. We also find that phase asymmetry controls the probability of occurrence of particular routes to synchronization. Our results suggest that asymmetry plays a crucial role in controlling synchronization within and between oscillator ensembles, and hence that its consideration is vital for modeling real life problems.


Behavioral, situational, and temporal effects of treatment of ADHD with methylphenidate.
To determine the behavioral, situational, and temporal effects of 4 months of methylphenidate (MPH) treatment for attention-deficit hyperactivity disorder (ADHD). Ninety-one children with ADHD were randomly assigned to receive either MPH (titrated to a target dose of 0.7 mg/kg twice a day) or a placebo. Treatment effects were investigated with measures sensitive to various behaviors (core and associated symptoms), situations (home and school), time periods (morning and afternoon, after reaching the target dose, and after 4 months of treatment), and side effects. MPH treatment improved symptoms of ADHD and oppositional behavior at school, both in the morning and afternoon, but not at home. Side effects (increase in physiological and effective symptoms, lack of weight gain) were significantly more frequent with MPH than with placebo treatment. Benefit was evident after titration, but the onset of some side effects was delayed. Side effects were reported by parents but not by teachers. Positive effects of MPH on behavior are evident in the classroom, but with MPH given twice daily, parents do not report that MPH improves behavior at home. Greater impact on home behavior may require three times daily MPH and combined treatments.


Advances in using Internet searches to track dengue.
Dengue is a mosquito-borne disease that threatens over half of the world's population. Despite being endemic to more than 100 countries, government-led efforts and tools for timely identification and tracking of new infections are still lacking in many affected areas. Multiple methodologies that leverage the use of Internet-based data sources have been proposed as a way to complement dengue surveillance efforts. Among these, dengue-related Google search trends have been shown to correlate with dengue activity. We extend a methodological framework, initially proposed and validated for flu surveillance, to produce near real-time estimates of dengue cases in five countries/states: Mexico, Brazil, Thailand, Singapore and Taiwan. Our result shows that our modeling framework can be used to improve the tracking of dengue activity in multiple locations around the world.


Protein kinase cα regulates the expression of complement receptor Ig in human monocyte-derived macrophages.
The complement receptor Ig (CRIg) is selectively expressed by macrophages. This receptor not only promotes the rapid phagocytosis of bacteria by macrophages but also has anti-inflammatory and immunosuppressive functions. Previous findings have suggested that protein kinase C (PKC) may be involved in the regulation of CRIg expression in human macrophages. We have now examined the role of PKCα in CRIg expression in human monocyte-derived macrophages (MDM). Macrophages nucleofected with plasmid containing short hairpin RNA against PKCα showed markedly reduced expression of PKCα, but normal PKCζ expression, by Western blotting analysis, and vice versa. PKCα-deficient MDM showed increased expression of CRIg mRNA and protein (both the long and short form), an increase in phagocytosis of complement-opsonized Candida albicans, and decreased production of TNF-α and IL-6. TNF-α caused a marked decrease in CRIg expression, and addition of anti-TNF mAb to the TNF-α-producing MDMs increased CRIg expression. PKCα-deficient macrophages also showed significantly less bacterial LPS-induced downregulation of CRIg. In contrast, cells deficient in PKCα showed decreased expression of CR type 3 (CR3) and decreased production of TNF-α and IL-6 in response to LPS. MDM developed under conditions that increased expression of CRIg over CR3 showed significantly reduced production of TNF-α in response to opsonized C. albicans. The findings indicate that PKCα promotes the downregulation of CRIg and upregulation of CR3 expression and TNF-α and IL-6 production, a mechanism that may promote inflammation.


Emergent social cohesion for coping with community disruptions in disasters.
Social cohesion is an important determinant of community well-being, especially in times of distress such as disasters. This study investigates the phenomena of emergent social cohesion, which is characterized by abrupt, temporary and extensive social ties with the goal of sharing and receiving information regarding a particular event influencing a community. In the context of disasters, emergent social cohesion, enabled by social media usage, could play a significant role in improving the ability of communities to cope with disruptions in recent disasters. In this study, we employed a network reticulation framework to examine the underlying mechanisms influencing emergent social cohesion on social media while communities cope with disaster-induced disruptions. We analysed neighbourhood-tagged social media data (social media data whose users are tagged by neighbourhoods) in Houston, TX, USA, during Hurricane Harvey to characterize four modalities of network reticulation (i.e. enactment, activation, reticulation and performance) giving rise to emergent social cohesion. Our results show that, unlike regular social cohesion, communication history and physical proximity do not significantly affect emergent social cohesion. The results also indicate that weak social ties play an important role in bridging different social network communities, and hence reinforce emergent social cohesion. The findings can inform public officials, emergency managers and decision-makers regarding the important role of neighbourhood-tagged social media, as a new form of community infrastructure, for improving the ability of communities to cope with disaster disruptions through enhanced emergent social cohesion.


Spatial and temporal distribution of polyamine levels and polyamine anabolism in different organs/tissues of the tobacco plant. Correlations with age, cell division/expansion, and differentiation.
Polyamine (PA) titers and biosynthesis follow a basipetal decrease along the tobacco (Nicotiana tabacum) plant axis, and they also correlate negatively with cell size. On the contrary, the titers of arginine (Arg), ornithine (Orn), and arginase activity increase with age. The free (soluble)/total-PA ratios gradually increase basipetally, but the soluble conjugated decrease, with spermidine (Spd) mainly to determine these changes. The shoot apical meristems are the main site of Spd and spermine biosynthesis, and the hypogeous tissues synthesize mostly putrescine (Put). High and low Spd syntheses are correlated with cell division and expansion, respectively. Put biosynthetic pathways are differently regulated in hyper- and hypogeous tobacco tissues: Only Arg decarboxylase is responsible for Put synthesis in old hypergeous vascular tissues, whereas, in hypogeous tissues, arginase-catalyzed Orn produces Put via Orn decarboxylase. Furthermore, Orn decarboxylase expression coincides with early cell divisions in marginal sectors of the lamina, and Spd synthase strongly correlates with later cell divisions in the vascular regions. This detailed spatial and temporal profile of the free, soluble-conjugated, and insoluble-conjugated fractions of Put, Spd, and spermine in nearly all tobacco plant organs and the profile of enzymes of PA biosynthesis at the transcript, protein, and specific activity levels, along with the endogenous concentrations of the precursor amino acids Arg and Orn, offer new insight for further understanding the physiological role(s) of PAs. The results are discussed in the light of age dependence, cell division/expansion, differentiation, phytohormone gradients, senescence, and sink-source relationships.


Chitinimonas lacunae sp. nov., isolated from artificial pond in Korea.
A gram-negative, rod shaped bacterium designated as strain H2T was isolated from an artificial pond in Korea. The strain H2T was able to grow aerobically and anaerobically with optimal growth occurring at 30 °C and pH 7.0 under aerobic conditions. Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain H2T belonged to the genus Chitinimonas of the family Burkholderiaceae. Phylogenetic similarity calculated from 16S rRNA gene sequences of strain H2T and valid species belongs to the genus Chitinimonas ranged from 93.2 % (for Chitinimonas taiwanensis cfT) to 94.4 % (for Chitinimonas prasina LY03T), and strain H2T formed a tight monophyletic group with them. Predominant fatty acids were C16 : 0 and summed feature 3, which consisted of C16 : 1ω6c and/or C16 : 1ω7c. The major respiratory quinone of the strain H2T was ubiquinone-8, and DNA G+C content was 60.2 %. The polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, unidentified aminolipid, and unidentified phospholipid. The biochemical characteristics that distinguished strain H2T from other Chitinimonas species included positive cystine arylamidase activity and lacked α-chymotrypsin and β-glucosidase (aesculin hydrolysis) activity. In addition, reciprocal DNA-DNA relatedness between H2T and three Chitinimonas strains ranged from 32.0 to 43.7 %. On the basis of its phylogenetic, chemotaxonomic, and genotypic characteristics, strain H2T represents a novel species of the genus Chitinimonas. Chitinimonas lacunae sp. nov. is proposed with the type strain H2T (=KCTC 52574T=LMG 29894T).


Antisense GLUT-1 protects mesangial cells from glucose induction of GLUT-1 and fibronectin expression.
A stable clone of rat mesangial cells expressing antisense GLUT-1 (i.e., MCGT1AS cells) was developed to protect them from high glucose exposure. GLUT-1 protein was reduced 50%, and the 2-deoxy-[(3)H]glucose uptake rate was reduced 33% in MCGT1AS. MCLacZ control cells and MCGT1 GLUT-1-overexpressing cells were used for comparisons. In MCLacZ, 20 mM D-glucose increased GLUT-1 transcription 90% vs. no increase in MCGT1AS. Glucose (8 mM) and 12 mM xylitol [a hexose monophosphate (HMP) shunt substrate] did not stimulate GLUT-1 transcription. An 87% replacement of the standard 8 mM D-glucose with 3-O-methylglucose reduced GLUT-1 transcription 80%. D-Glucose (20 mM) increased fibronectin mRNA and protein by 47 and 100%, respectively, in MCLacZ vs. no increases in MCGT1AS. Fibronectin synthesis was elevated 48% in MCGT1 and reduced 44% in MCGT1AS. We conclude that 1) transcription of GLUT-1 in response to D-glucose depends on glucose metabolism, although not through the HMP shunt, and 2) antisense GLUT-1 treatment of mesangial cells blocks D-glucose-induced GLUT-1 and fibronectin expression, thereby demonstrating a protective effect that could be beneficial in the setting of diabetes.


Accuracy of Smartphone Images of the Cervix After Acetic Acid Application for Diagnosing Cervical Intraepithelial Neoplasia Grade 2 or Greater in Women With Positive Cervical Screening: A Systematic Review and Meta-Analysis.
Smartphones are used in cervical screening for visual inspection after acetic acid or Lugol's iodine (VIA/VILI) application to capture and share images to improve the sensitivity and interobserver variability of VIA/VILI. We undertook a systematic review and meta-analysis assessing the diagnostic accuracy of smartphone images of the cervix at the time of VIA/VILI (termed S-VIA) in the detection of precancerous lesions in women undergoing cervical screening. This systematic review was conducted in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Studies from January 1, 2010, to June 30, 2020, were assessed. MEDLINE/PubMed, Embase, CINAHL, Cochrane, and LILACS were searched. Cohort and cross-sectional studies were considered. S-VIA was compared with the reference standard of histopathology. We excluded studies where additional technology was added to the smartphone including artificial intelligence, enhanced visual assessment, and other algorithms to automatically diagnose precancerous lesions. The primary outcome was the accuracy of S-VIA for the diagnosis of cervical intraepithelial neoplasia grade 2 or greater (CIN 2+). Data were extracted, and we plotted the sensitivity, specificity, negative predictive value, and positive predictive value of S-VIA using forest plots. This study was prospectively registered with The International Prospective Register of Systematic Reviews:CRD42020204024. Six thousand three studies were screened, 71 full texts assessed, and eight studies met criteria for inclusion, with six included in the final meta-analysis. The sensitivity of S-VIA for the diagnosis of CIN 2+ was 74.56% (95% CI, 70.16 to 78.95; I2 61.30%), specificity was 61.75% (95% CI, 56.35 to 67.15; I2 95.00%), negative predictive value was 93.71% (95% CI, 92.81 to 94.61; I2 0%), and positive predictive value was 26.97% (95% CI, 24.13 to 29.81; I2 61.3%). Our results suggest that S-VIA has accuracy in the detection of CIN 2+ and may provide additional support to health care providers delivering care in low-resource settings.


Kinetic Signature of Cooperativity in the Irreversible Collapse of a Polymer.
We investigate the kinetics of a polymer collapse due to the formation of irreversible cross-links between its monomers. Using the contact probability P(s) as a scale-dependent order parameter depending on the chemical distance s, our simulations show the emergence of a cooperative pearling instability. Namely, the polymer undergoes a sharp conformational transition to a set of absorbing states characterized by a length scale ξ corresponding to the mean pearl size. This length and the transition time depend on the polymer equilibrium dynamics and the cross-linking rate. We confirm experimentally this transition using a DNA conformation capture experiment in yeast.


Evaluation of portable blood glucose meters. Problems and recommendations.
Various guidelines are in use for validating blood glucose meters. There are currently no analytical guidelines for validating procedures available for non-invasive or minimally invasive blood glucose meters. As a result manufacturers of these meters need to comply with different regulations in different countries with concomitant high costs. Some of the differences between different guidelines will be discussed in this article and recommendations for improvement will be presented.


A NEW REGISTRATION METHOD BASED ON LOG-EUCLIDEAN TENSOR METRICS AND ITS APPLICATION TO GENETIC STUDIES.
In structural brain MRI, group differences or changes in brain structures can be detected using Tensor-Based Morphometry (TBM). This method consists of two steps: (1) a non-linear registration step, that aligns all of the images to a common template, and (2) a subsequent statistical analysis. The numerous registration methods that have recently been developed differ in their detection sensitivity when used for TBM, and detection power is paramount in epidemological studies or drug trials. We therefore developed a new fluid registration method that computes the mappings and performs statistics on them in a consistent way, providing a bridge between TBM registration and statistics. We used the Log-Euclidean framework to define a new regularizer that is a fluid extension of the Riemannian elasticity, which assures diffeomorphic transformations. This regularizer constrains the symmetrized Jacobian matrix, also called the deformation (or strain) tensor. We applied our method to an MRI dataset from 40 fraternal and identical twins, to revealed voxelwise measures of average volumetric differences in brain structure for subjects with different degrees of genetic resemblance.


Structures that delineate orphanin FQ and dynorphin A pharmacological selectivities.
Strict pharmacological selectivity in families of structurally related ligands and receptors may result from a key process in evolution aiming at increasing diversity in neurotransmission. An intriguing example of such exclusive specificity can be found in the newly discovered orphanin FQ (OFQ) system when it is compared with the opioid system. Both OFQ and its receptor share a high degree of sequence similarity to the opioid peptides and their corresponding receptors, respectively. However, OFQ does not activate opioid receptors, nor do the opioid peptides elicit biological activity at the OFQ receptor. We have therefore investigated the basis for the inherent selectivity of the primary structures of OFQ and dynorphin A, its closest counterpart. A series of truncated and/or chimeric peptides led to the conclusion that both peptides contain domains which establish their pharmacological selectivity. In the OFQ molecule we could delineate a domain that prevents its ability to activate the kappa-opioid receptor by apparently repelling its binding. In both peptides the selectivity-generating domains are composed of single residues in key positions together with short stretches of amino acids which do not overlap. To prove this concept, we designed a universal agonist and found it active at both the OFQ receptor and the kappa-opioid receptor. Our observations suggest that a coordinated mechanism of evolution has separated the orphanin FQ system from the opioid system.


Recommendations to enable drug development for inherited neuropathies: Charcot-Marie-Tooth and Giant Axonal Neuropathy.
Approximately 1 in 2500 Americans suffer from Charcot-Marie-Tooth (CMT) disease. The underlying disease mechanisms are unique in most forms of CMT, with many point mutations on various genes causing a toxic accumulation of misfolded proteins. Symptoms of the disease often present within the first two decades of life, with CMT1A patients having reduced compound muscle and sensory action potentials, slow nerve conduction velocities, sensory loss, progressive distal weakness, foot and hand deformities, decreased reflexes, bilateral foot drop and about 5% become wheelchair bound. In contrast, the ultra-rare disease Giant Axonal Neuropathy (GAN) is frequently described as a recessively inherited condition that results in progressive nerve death. GAN usually appears in early childhood and progresses slowly as neuronal injury becomes more severe and leads to death in the second or third decade. There are currently no treatments for any of the forms of CMTs or GAN. We suggest that further clinical studies should analyse electrical impedance myography as an outcome measure for CMT. Further, additional quality of life (QoL) assessments for these CMTs are required, and we need to identify GAN biomarkers as well as develop new genetic testing panels for both diseases. We propose that using the Global Registry of Inherited Neuropathy (GRIN) could be useful for many of these studies. Patient advocacy groups and professional organizations (such as the Hereditary Neuropathy Foundation (HNF), Hannah's Hope Fund (HHF), The Neuropathy Association (TNA) and the American Association of Neuromuscular and Electrodiagnostic Medicine (AANEM) can play a central role in educating clinicians and patients. Undertaking these studies will assist in the correct diagnosis of disease recruiting patients for clinical studies, and will ultimately improve the endpoints for clinical trials. By addressing obstacles that prevent industry investment in various forms of inherited neuropathies, we can envision treatment options for these rare diseases in the near future.