Synthesis of Human Bone Morphogenetic Protein-2 (hBMP-2) in E. coli Periplasmic Space: Its Characterization and Preclinical Testing.
Human BMP-2, a homodimeric protein that belongs to the TGF- β family, is a recognized osteoinductor due to its capacity of inducing bone regeneration and ectopic bone formation. The administration of its recombinant form is an alternative to autologous bone grafting. A variety of E. coli-derived hBMP-2 has been synthesized through refolding of cytoplasmic inclusion bodies. The present work reports the synthesis, purification, and characterization of periplasmic hBMP-2, obtained directly in its correctly folded and authentic form, i.e., without the initial methionine typical of the cytoplasmic product that can induce undesired immunoreactivity. A bacterial expression vector was constructed including the DsbA signal peptide and the cDNA of hBMP-2. The periplasmic fluid was extracted by osmotic shock and analyzed via SDS-PAGE, Western blotting, and reversed-phase high-performance liquid chromatography (RP-HPLC). The purification was carried out by heparin affinity chromatography, followed by high-performance size-exclusion chromatography (HPSEC). HPSEC was used for qualitative and quantitative analysis of the final product, which showed >95% purity. The classical in vitro bioassay based on the induction of alkaline phosphatase activity in myoblastic murine C2C12 cells and the in vivo bioassay consisting of treating calvarial critical-size defects in rats confirmed its bioactivity, which matched the analogous literature data for hBMP-2.


Observations of comet 19P/Borrelly by the miniature integrated camera and spectrometer aboard Deep Space 1.
The nucleus of the Jupiter-family comet 19P/Borrelly was closely observed by the Miniature Integrated Camera and Spectrometer aboard the Deep Space 1 spacecraft on 22 September 2001. The 8-kilometer-long body is highly variegated on a scale of 200 meters, exhibiting large albedo variations (0.01 to 0.03) and complex geologic relationships. Short-wavelength infrared spectra (1.3 to 2.6 micrometers) show a slope toward the red and a hot, dry surface (</=345 kelvin, with no trace of water ice or hydrated minerals), consistent with approximately 10% or less of the surface actively sublimating. Borrelly's coma exhibits two types of dust features: fans and highly collimated jets. At encounter, the near-nucleus coma was dominated by a prominent dust jet that resolved into at least three smaller jets emanating from a broad basin in the middle of the nucleus. Because the major dust jet remained fixed in orientation, it is evidently aligned near the rotation axis of the nucleus.


[Somatotyping of children and adolescents].
Longitudinal observations on growth and development in children and adolescents with a usual (school) and increased (sporting) motor activity regimens have been performed for 13 years and certain metrical characteristics have been obtained. After their treatment by means of the factor and correlative analyses, they are divided into basic and auxiliary. The basic characteristics have a high correlative connection with progress in certain kinds of physical activity (sporting achievements), the auxiliary ones determine the technique of carrying out the movement. Basing on theoretical generalization of the literature data and analyzing the metrical values obtained at anthropometry, an objective scheme of somatotyping is worked out. It considers an independent variation of the signs according to spatial (clearance limit, componental and proportional) and time (variants of development) scales. Nontraditional estimation on manifestation of fatty, muscular and osseous components is elaborated, algorhythm of their estimation is suggested. Three main (micro-, meso-, macrosomatic) and two additional (micromesosomatic and mesomacrosomatic) somatic types are defined. They include manifestation of fatty, muscular and osseous masses, as well as the proportion of the body-build. The mark scale estimation of individual metrical values according to the elaborated age nomograms and scales makes the base of the typing. As demonstrates the retrospective analysis of the material, in 86% of cases the somatic type does not change in the course of ontogenesis. Estimation of the somatic type during the pubertal period should be carried out with a strict consideration of the variant of the child's development.(ABSTRACT TRUNCATED AT 250 WORDS)


Laparoscopic debridement of hepatic necrosis after hepatic artery chemoembolization.
Hepatic artery chemoembolization (HACE) used to treat neuroendocrine tumors metastatic to the liver has shown both survival benefit and improvement in symptoms. The development of hepatic necrosis after HACE is rare, but the consequences can be devastating. We report the first case of laparoscopic management of extensive hepatic necrosis occurring after HACE. A 58-year-old man with neuroendocrine tumor metastatic to the liver underwent HACE in addition to medical management. He had undergone previous biliary stenting for biliary obstruction. After HACE was performed via the right hepatic artery, the patient developed sepsis due to right hepatic lobe infarction. Percutaneous drainage and antibiotics were attempted for 2 months, but hepatic debridement was ultimately required due to repeated drain malfunction and septic complications. Laparoscopic necrosectomy was performed with ease and with little blood loss. The patient quickly recovered without any further infectious complications. Infected hepatic necrosis resulting from HACE that fails percutaneous management can be successfully managed with laparoscopic necrosectomy. This report adds to the growing evidence that minimally invasive techniques can be used to manage complicated hepatic conditions.


Burs, teeth and hand instruments.
Scanning electron microscope studies demonstrate deficiencies in the surfaces of cavity preparations made with conventional cutting instruments. Better surface preparation can be achieved by using appropriate shaped tungsten rotary instruments, and by sharp hand instruments used with a scraping action. The direction of rotation of the cutting alters the quality of the enamel margin. The SEM image should not be interpreted as the sole criterion of clinical success.


Improved sleep, cognitive processing and enhanced learning and memory task accuracy with Yoga nidra practice in novices.
Complementary and Alternative medicine is known to have health benefits. Yoga nidra practice is an easy-to-do practice and has shown beneficial effects on stress reduction and is found to improve sleep in insomnia patients. Effect of yoga nidra practice on subjective sleep is known but its effect on sleep and cognition objectively is not documented. The aim of the study was to study the effect of yoga nidra practice on cognition and sleep using objective parameters. 41 participants were enrolled, and baseline sleep diary (SD) collected. Participants volunteered for overnight polysomnography (PSG) and cognition testing battery (CTB) comprising of Motor praxis test, emotion recognition task (ERT), digital symbol substitution task, visual object learning task (VOLT), abstract matching (AIM), line orientation task, matrix reasoning task, fractal-2-back test (NBACK), psychomotor vigilance task and balloon analog risk task. Baseline CTB and after one and two weeks of practice was compared. Power spectra density for EEG at central, frontal, and occipital locations during CTB was compared. Repeat SD and PSG after four weeks of practice were done. After yoga nidra practice, improved reaction times for all cognition tasks were seen. Post intervention compared to baseline (95%CI; p-value, effect size) showed a significant improvement in sleep efficiency of +3.62% (0.3, 5.15; p = 0.03, r = 0.42), -20min (-35.78, -5.02; p = 0.003, d = 0.84) for wake after sleep onset and +4.19 μV2 (0.5, 9.5; p = 0.04, r = 0.43) in delta during deep sleep. Accuracy increased in VOLT (95% CI: 0.08, 0.17; p = 0.002, d = 0.79), AIM (95% CI: 0.03, 0.12; p = 0.02, d = 0.61) and NBACK (95% CI: 0.02, 0.13; p = 0.04, d = 0.56); ERT accuracy increased for happy, fear and anger (95% CI: 0.07, 0.24; p = 0.004, d = 0.75) but reduced for neutral stimuli (95% CI: -0.31, -0.12; p = 0.04, r = 0.33) after yoga nidra practice. Yoga Nidra practice improved cognitive processing and night-time sleep.


T lymphocyte tolerance and early appearance of virus-induced cell surface antigens in Moloney-murine leukemia virus neonatally injected mice.
Regression of Moloney-murine sarcoma virus- (M-MSV) induced sarcomas in normal adult mice is accompanied by generation of virus-specific cytotoxic T lymphocytes (CTL). However, when neonatal mice that were injected with Moloney-murine leukemia virus (M-MuLV carrier) were subsequently challenged as adults with M-MSV, the sarcomas did not regress nor did they generate CTL. This failure to produce CTL cannot be ascribed to nonspecific immunodepressive effects or to suppressor cell generation since M-MuLV carrier mice exhibit normal reactivity after allogeneic cell stimulation. Moreover, addition of M-MuLV-infected cells as the third party to cultures does not reduce activity of CTL from M-MSV immune mice. Since M-MSV and M-MuLV possess common antigens, the observed unresponsiveness was considered in relationship to induction of a T lymphocyte tolerance, which may follow introduction of foreign antigens at an early stage of development. In fact, it was observed that as early as 10 days after injection, thymus, lymph node, and spleen from M-MuLV carrier mice express virus-induced cell-surface antigens that not only are targets for M-MSV-immune CTL, but also induce in vitro a strong specific cytotoxic response. In addition, a cold target inhibition assay disclosed that the same antigens are shared by both M-MuLV infected and leukemia cells, even though they are less expressed on the surface of the former. The finding that the cytotoxicity of alloreactive lymphocytes from M-MuLV carrier mice is reduced after preincubation with M-MSV immune CTL confirms that virus infection does not bring about functional inactivation of lymphocytes. Finally, it was observed that virus antigen presence on lymphocytes from M-MuLV neonatally injected mice is closely related to subsequent leukemia development.


Characteristics of storm runoff and sediment dispersal in the San Pedro Channel, southern California.
In-site measurements of particle size spectra were obtained from three offshore cruises to evaluate the physical consequences of increased sediment transport and deposition offshore which was caused by episodic storm runoff water from the Santa Ana River watershed, a highly urbanised coastal watershed in southern California. Of the total annual runoff discharge to the coastal ocean, 89.2% occurred in the 2003/2004 winter season, and 0.22 Mt of sediment mass was transported during the storm events. The runoff plume at surface taken offshore by cross-shore currents progressed rapid aggregation and sedimentation, while the initially high concentration of suspended sediment discharged from the river outlet was dominated by small particles. Vertical profiles of particle size spectra revealed two separated plumes near the river outlet and turbidity plume along the bottom consisted of an abundance of very fine and dense particles. It would appear to support the theory that even if the storm runoff does not carry a high concentration of sediment being capable of generating negative buoyancy, sediment deposition on the shelf might mobilise in dense, fluid mud transported offshore by gravity. In a coastal pollution context, sediment particle size spectra information may offer potentially useful means of characterising particle-associated pollutants for purposes of source tracking and environmental interpretation.


Tumor growth rate as a metric of progression, response, and prognosis in pancreatic and intestinal neuroendocrine tumors.
Lanreotide depot/autogel antitumor activity in intestinal/pancreatic neuroendocrine tumors (NETs) was demonstrated in the phase-3 CLARINET study (NCT00353496), based on significantly prolonged progression-free survival (PFS) versus placebo. During CLARINET, patients with metastatic intestinal/pancreatic NETs received lanreotide depot/autogel 120 mg or placebo every 4 weeks for 96 weeks. Imaging data (response evaluation criteria in solid tumors [RECIST] v1.0, centrally reviewed) were re-evaluated in this post hoc analysis of tumor growth rate (TGR) in NETs. TGR (%/month) was calculated from two imaging scans during relevant periods: pre-treatment (TGR0); 12-24 weeks before randomization versus baseline; each treatment visit versus baseline (TGRTx-0); between consecutive treatment visits (TGRTx-Tx). To assess TGR as a measure of prognosis, PFS was compared for TGR0 subgroups stratified by optimum TGR0 cut-off; a multivariate analysis was conducted to identify prognostic factors for PFS. TGR0 revealed tumors growing during pre-treatment (median [interquartile range] TGR0: lanreotide 2.1%/month [0.2; 6.1]; placebo 2.7%/month [0.15; 6.8]), contrary to RECIST status. TGR was significantly reduced by 12 weeks with lanreotide versus placebo (difference in least-square mean TGR0-12 of - 2.9 [- 5.1, - 0.8], p = 0.008), a difference that was maintained at most subsequent visits. TGR0 > 4%/month had greater risk of progression/death than ≤4%/month (hazard ratio 4.1; [95% CI 2.5-6.5]; p < 0.001); multivariate analysis revealed lanreotide treatment, progression at baseline, TGR0, hepatic tumor load, and primary tumor type were independently associated with PFS. TGR provides valuable information on tumor activity and prognosis in patients with metastatic intestinal/pancreatic NETs, and identifies early lanreotide depot/autogel antitumor activity. Retrospective registration, 18 July 2006; EudraCT: 2005-004904-35; ClinicalTrials.gov: NCT00353496 .


Serine/threonine phosphorylation of calmodulin modulates its interaction with the binding domains of target enzymes.
The interaction of serine/threonine-phosphorylated calmodulin with synthetic peptides corresponding to the calmodulin-binding domains of six enzymes has been studied by fluorescence spectroscopy. For five peptides, the dissociation constant of the calmodulin-peptide complex (K(d)) increased when calmodulin was phosphorylated. An increase of more than one order of magnitude was observed with peptides derived from smooth-muscle myosin light-chain kinase and cAMP phosphodiesterase. In contrast, only a slight increase in K(d) was noted with two peptides derived from the plasma membrane Ca(2+)-ATPase and for the peptide derived from nitric oxide synthase. No significant change in affinity was detected with the peptide derived from calcineurin. In contrast, a decrease in the dissociation constant was observed with the peptide derived from the Ca(2+)-calmodulin dependent kinase II. Phosphorylation also affected the peptide-calmodulin binding stoichiometry: a decrease from two to one binding sites was observed with the peptides derived from myosin light-chain kinase and phosphodiesterase.


Femtosecond optical parametric oscillator continuously tunable across 3.6-8 μm based on orientation-patterned gallium phosphide.
We report a synchronously-pumped femtosecond optical parametric oscillator (OPO) based on orientation-patterned gallium phosphide (OP-GaP), providing continuously tunable mid-infrared (mid-IR) idler radiation across 3570-7892 nm (2801-1267 cm-1), by exploiting pump wavelength tuning. We generate up to 54 mW of output average power at 80 MHz, and quantum conversion efficiencies up to 28.9% are achieved by use of synchronized pump retroreflection. With the inclusion of intracavity dispersion compensation in an OP-GaP OPO, near-transform-limited signal pulse durations of 112 fs at 1288 nm are measured, and peak powers up to 3 kW in the mid-IR are inferred. Finally, evidence of three-photon absorption is observed and characterized near the pump wavelength.


[The role of direct-acting P2Y12 inhibitors in acute coronary syndrome].
P2Y12 receptor inhibitors are recommended for treatment of patients with acute coronary syndrome and/or percutaneous coronary intervention as a part of dual antiplatelet therapy. While thienopyridine group of P2Y12 inhibitors remains to be widely used treatment for patients with ACS, there are still complications for its application. Emerging antiplatelet treatments expand therapy strategies for management of ACS. The purpose of this article is to define the developing role of novel non-thienopyridine reversible P2Y12 inhibitor ticagrelor to present its pharmacokinetic, pharmacodynamic, and pharmacogenetic characteristics, based on current evidence.


Effect of Herbst treatment on temporomandibular joint morphology: a systematic literature review.
The purpose of this systematic review was to evaluate the effect of Herbst appliance therapy on temporomandibular joint (TMJ) morphology, with special reference to glenoid fossa remodeling, condylar remodeling, condylar position, and articular disc position. Publications of controlled trials of Herbst treatment of Class II patients using magnetic resonance imaging (MRI), computerized tomography scans, or tomography to assess TMJ morphology were identified with Medline (1966-2001), Best Evidence (1991-2001), Cochrane Database of Systematic Reviews (second quarter, 2001), and Embase (1998-2001). Case reports were excluded. Based on our search, only 5 studies met the selection criteria. All studies used internal controls with pretreatment and posttreatment imaging. Four studies used MRI, and 1 used tomograms. The 4 MRI studies used overlapping patient samples and were not considered as independent evidence. The MRI studies did not provide conclusive evidence of osseous remodeling or condyle position change. The tomography study demonstrated minor condyle position change. Methodological deficiencies prevented major conclusions regarding disc position. The reviewed studies highlight the importance of further research. Prospective controlled studies using serial MRI and tomography are required to establish the effect of Herbst treatment on TMJ morphology.


Studies on long-term depression in area CA1 of the anesthetized and freely moving rat.
Homosynaptic long-term depression (LTD) is reported to occur in field CA1 of hippocampal slices collected from immature brains. Because the effect has been postulated to be a memory storage mechanism, it is of interest to test for its presence in adult, awake animals. Unfortunately, not only has hippocampal LTD proved difficult to obtain reliably in vivo, but the few successful studies vary with respect to protocols and evidence that the depression is input-specific. The present study tested for input-specific (homosynaptic) LTD in field CA1 after application of various stimulation protocols to the Schaffer collateral/commissural projections in freely moving, adult rats. The results indicate that although low-frequency trains do induce decrements in synaptic transmission lasting for hours to several days, the success rate of eliciting input-specific LTD in the awake rat is very modest compared with the ease with which stable potentiation is obtained in the same synapses. Moreover, it is questionable that the effective protocols represent patterns of activity likely to occur during behavior. The stronger the afferent activation during low-frequency stimulation, the greater was the probability of eliciting LTD accompanied by persistent heterosynaptic depression. Clear evidence for the occurrence of LTD, irrespective of stimulation protocol and current intensity, could not be obtained in rats under barbiturate anesthesia. In all, the results do not accord with the suggestion that LTD occurs routinely in the hippocampus in vivo as part of memory encoding.


Strong Costs and Benefits of Winter Acclimatization in Drosophila melanogaster.
Studies on thermal acclimation in insects are often performed on animals acclimated in the laboratory under conditions that are not ecologically relevant. Costs and benefits of acclimation responses under such conditions may not reflect costs and benefits in natural populations subjected to daily and seasonal temperature fluctuations. Here we estimated costs and benefits in thermal tolerance limits in relation to winter acclimatization of Drosophila melanogaster. We sampled flies from a natural habitat during winter in Denmark (field flies) and compared heat and cold tolerance of these to that of flies collected from the same natural population, but acclimated to 25 °C or 13 °C in the laboratory (laboratory flies). We further obtained thermal performance curves for egg-to-adult viability of field and laboratory (25 °C) flies, to estimate possible cross-generational effects of acclimation. We found much higher cold tolerance and a lowered heat tolerance in field flies compared to laboratory flies reared at 25 °C. Flies reared in the laboratory at 13 °C exhibited the same thermal cost-benefit relations as the winter acclimatized flies. We also found a cost of winter acclimatization in terms of decreased egg-to-adult viability at high temperatures of eggs laid by winter acclimatized flies. Based on our findings we suggest that winter acclimatization in nature can induce strong benefits in terms of increased cold tolerance. These benefits can be reproduced in the laboratory under ecologically relevant rearing and testing conditions, and should be incorporated in species distribution modelling. Winter acclimatization also leads to decreased heat tolerance. This may create a mismatch between acclimation responses and the thermal environment, e.g. if temperatures suddenly increase during spring, under current and expected more variable future climatic conditions.


Bioactivity assessment of natural compounds using machine learning models trained on target similarity between drugs.
Natural compounds constitute a rich resource of potential small molecule therapeutics. While experimental access to this resource is limited due to its vast diversity and difficulties in systematic purification, computational assessment of structural similarity with known therapeutic molecules offers a scalable approach. Here, we assessed functional similarity between natural compounds and approved drugs by combining multiple chemical similarity metrics and physicochemical properties using a machine-learning approach. We computed pairwise similarities between 1410 drugs for training classification models and used the drugs shared protein targets as class labels. The best performing models were random forest which gave an average area under the ROC of 0.9, Matthews correlation coefficient of 0.35, and F1 score of 0.33, suggesting that it captured the structure-activity relation well. The models were then used to predict protein targets of circa 11k natural compounds by comparing them with the drugs. This revealed therapeutic potential of several natural compounds, including those with support from previously published sources as well as those hitherto unexplored. We experimentally validated one of the predicted pair's activities, viz., Cox-1 inhibition by 5-methoxysalicylic acid, a molecule commonly found in tea, herbs and spices. In contrast, another natural compound, 4-isopropylbenzoic acid, with the highest similarity score when considering most weighted similarity metric but not picked by our models, did not inhibit Cox-1. Our results demonstrate the utility of a machine-learning approach combining multiple chemical features for uncovering protein binding potential of natural compounds.


Using Certification to Promote Uptake of Real-World Evidence by Payers.
Most randomized controlled trials are unable to generate information about a product's real-world effectiveness. Therefore, payers use real-world evidence (RWE) generated in observational studies to make decisions regarding formulary inclusion and coverage. While some payers generate their own RWE, most cautiously rely on RWE produced by manufacturers who have a strong financial interest in obtaining coverage for their products. We propose a process by which an independent body would certify observational studies as generating valid and unbiased estimates of the effectiveness of the intervention under consideration. This proposed process includes (a) establishing transparent criteria for assessment, (b) implementing a process for receipt and review of observational study protocols from interested parties, (c) reviewing the submitted protocol and requesting any necessary revisions, (d) reviewing the study results, (e) assigning a certification status to the submitted evidence, and (f) communicating the certification status to all who seek to use this evidence for decision making. Accrediting organizations such as the National Center for Quality Assurance and the Joint Commission have comparable goals of providing assurance about quality to those who look to their accreditation results. Although we recognize potential barriers, including a slowing of evidence generation and costs, we anticipate that processes can be streamlined, such as when familiar methods or familiar datasets are used. The financial backing for such activities remains uncertain, as does identification of organizations that might serve this certification function. We suggest that the rigor and transparency that will be required with such a process, and the unassailable evidence that it will produce, will be valuable to decision makers.


Applications of microwave, ultrasound, and radiofrequency heating.
Different needs must be considered in the control of localized tumors or disseminated disease when deep heating techniques are used. For localized disease, various electromagnetic microwaves, radiofrequency currents and high-powered ultrasound sources are being used. At the present time, each approach has its own strengths and limitations. Microwave and inductive approaches appear to be useful only for surface lesions. Radiofrequency currents, generated by pairs of plates, give more deeper heating in the body, but excessive heating occurs in fatty layers related to high resistivity of the tissue and sometimes near the edge of the applicators. In an invasive approach, needles are implanted into the tumor and excited by a radiofrequency source. Considerable clinical successes are claimed for local heating of the lesion with this approach, but it is only useful for specific sites of the body. Ultrasound has special advantages for both surface or deep-seated soft tissues due to its flexibility and its ability to focus the beam onto the lesion. However, ultrasound cannot be used in the chest cavity or near bone, and the dosimetry is not defined at present. For disseminated disease, much larger regions, particularly in the chest cavity, can be heated, with the hope that the reduced blood circulation in some tumors can achieve a preferential heating in the lesion. Several microwave and inductive heating approaches are being used or considered in the achievement of this goal.


The interferon-gamma gene as a positional and functional candidate gene for inflammatory bowel disease.
Epidemiological and genome-wide linkage analyses have provided firm evidence for a genetic component in the pathogenesis of inflammatory bowel disease. The linkage regions on chromosomes 12 and 16 have been replicated in several independent samples. These represent the best positional evidence in the search for inflammatory bowel disease susceptibility genes. While systematic association and physical mapping studies in these regions are under way, the direct analysis of immunologically relevant genes as positional and functional candidates may provide a shortcut in this process. The interferon-gamma gene resides in the chromosome 12 linkage region near the marker D12S83. Interferon-gamma is an important proinflammatory cytokine in the interleukin-12 cascade and has been implicated in the pathogenesis of mucosal inflammation. We tested this gene for evidence of linkage and association in 133 German multiplex families and 506 single patients with their parents. An intragenic, highly informative CA-repeat marker in intron 1 of the gene was typed using fluorescence-labeled polymerase chain reaction and analysis on an automated sequencer. In the nonparametric linkage analysis using GENEHUNTER, a nonsignificant maximum LOD score of 0.67 was obtained. The transmission disequilibrium test for association was negative (P > or = 0.22) for Crohn's disease, ulcerative colitis, and the combined inflammatory bowel disease phenotype. In summary, the findings make interferon-gamma a very unlikely candidate for the major susceptibility gene in the chromosome 12 linkage interval. Future efforts can concentrate on other transcripts in the region.


Persistence of vesicular stomatitis virus in cloned interleukin-2-dependent natural killer cell lines.
We have investigated virus-lymphocyte interactions by using cloned subpopulations of interleukin-2-dependent effector lymphocytes maintained in vitro. Cloned lines of H-2-restricted hapten- or virus-specific cytotoxic T lymphocytes (CTL) and alloantigen-specific CTL were resistant to productive infection by vesicular stomatitis virus (VSV). In contrast, cloned lines of natural killer (NK) cells were readily and persistently infected by VSV, a virus which is normally highly cytolytic. VSV-infected NK cells continued to proliferate, express viral surface antigen, and produce infectious virus. Furthermore, persistently infected NK cells showed no marked alteration of normal cellular morphology and continued to lyse NK-sensitive target cells albeit at a slightly but significantly reduced level. The persistence of VSV in NK cells did not appear to be caused by the generation of temperature-sensitive viral mutants, defective interfering particles, or interferon. Consequently, studies comparing the intracellular synthesis and maturation of VSV proteins in infected NK and mouse L cells were conducted. In contrast to L cells, in which host cell protein synthesis was essentially totally inhibited by infection, the infection of NK cells caused no marked diminution in the synthesis of host cell proteins. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of immunoprecipitates of viral proteins from infected cells showed that the maturation rate and size of VSV surface G glycoprotein were comparable in L cells and NK cells. Nucleocapsid (N) protein synthesis also appeared to be unaffected in NK cells. In contrast, the viral proteins NS and M appeared to be selectively degraded in NK cell extracts. Mixing experiments suggested that a protease in NK cells was responsible for the selective breakdown of VSV NS protein. Finally, VSV-infected NK cells were resistant to lysis by virus-specific CTL, suggesting that persistently infected NK cells may harbor virus and avoid cell-mediated immune destruction in an immunocompetent host.


Study of Ground State Interactions of Enantiopure Chiral Quaternary Ammonium Salts and Amides, Nitroalkanes, Nitroalkenes, Esters, Heterocycles, Ketones and Fluoroamides.
Chiral phase-transfer catalysis provides high level of enantiocontrol, however no experimental data showed the interaction of catalysts and substrates. 1 H NMR titration was carried out on Cinchona and Maruoka ammonium bromides vs. nitro, carbonyl, heterocycles, and N-F containing compounds. It was found that neutral organic species and quaternary ammonium salts interacted via an ensemble of catalyst + N-C-H and (sp2 )C-H, specific for each substrate studied. The correspondent BArF salts interacted with carbonyls via a diverse set of + N-C-H and (sp2 )C-H compared to bromides. This data suggests that BArF ammonium salts may display a different enantioselectivity profile. Although not providing quantitative data for the affinity constants, the data reported proofs that chiral ammonium salts coordinate with substrates, prior to transition state, through specific C-H positions in their structures, providing a new rational to rationalize the origin of enantioselectivity in their catalyses.


Increased circulating microRNA-21 level as a potential indicator for predicting a higher risk of incident fragility fractures.
As a common disease in the elderly, osteoporosis clearly increases the risk of fractures, leading to higher mortality, but the current markers to estimate the risk of fractures are limited. MicroRNA-21 (miR-21) may play an important role in osteoporosis, but the link of this biomarker with fractures was undetermined. We aimed to investigate the association between miR-21 levels and the presence of fragility fractures. A total of 200 patients were recruited and miR-21 was collected from baseline serum. The correlation between miR-21 and the fracture risk assessment tool (FRAX) score was analyzed. The incidence of fragility fractures was presented by Kaplan-Meier analysis, and Cox regression analysis was utilized to evaluate risk factors. The diagnostic value of miR-21 was conducted by the area under curve (AUC). The FRAX score was significantly associated with miR-21 level (p<0.001). According to the 50th percentile of miR-21 content in the overall distribution, the cumulative incidence of fragility fractures was significantly higher in patients with higher miR-21 levels than those with lower levels (75.4, 95 % CI: 69.0-81.8 vs. 59.2, 95 % CI: 42.1-76.3, p<0.001). The results of the Cox regression analysis showed that the miR-21 level was an independent risk factor linked to the incidence of fracture (p=0.005). The optimal cut-off value of the miR-21 was 6.08, and the AUC for predicting fracture was 0.718 (95 % CI, 0.645-0.790). This study showed that miR-21 has optimal diagnostic performance in the discrimination of fragility fracture, and the circulating miR-21 level in predicting the risk of fragility fracture may have a certain value.


Enzyme-linked immunosorbent assay utilizing thin-layered microfluidics.
A rapid and sensitive enzyme-linked immunosorbent assay (ELISA) is required for on-site clinical diagnosis. Previously, a microfluidic ELISA in which antibody-immobilized beads are packed in a microchannel for a high surface-to-volume (S/V) ratio was developed, but utilizing beads led to complicated fluidic operation. Recently, we have reported nanofluidic ELISA that utilizes antibody-immobilized glass nanochannels (102-103 nm) to achieve a high S/V ratio without beads, enabling even single-molecule detection, but it is not applicable to clinical diagnosis owing to its fL sample volume, much smaller than the nL-μL sample volume in clinical diagnosis. Here, we propose an antibody-immobilized, thin-layered microfluidic channel as a novel platform. Based on the method of nanofluidic ELISA, the channel width was expanded from 103 nm to 100 mm to expand the volume of the reaction field to 102 nL, while the channel depth (103 nm) was maintained to retain the high S/V ratio. A device design which incorporates a taper-shaped interface between the thin-layered channel and the microchannel for sample injection was proposed, and the uniform introduction of the sample into the high-aspect-ratio (width/depth ∼ 200) channel was experimentally confirmed. For the proof of concept, a thin-layered ELISA device with the same S/V ratio as the bead-based ELISA format was designed and fabricated. By measuring a standard C-reactive protein solution, the working principle was verified. The limit of detection was 34 ng mL-1, which was comparable to that of bead-based ELISA. We believe that the thin-layered ELISA can contribute to medicine and biology as a novel platform for sensitive and rapid ELISA.


The war experiences and psychosocial development of children in Lebanon.
This study examines the number and types of war traumas children face growing up in a war-torn country and the relation of such traumatic experiences to their psychosocial development. A sample of 224 Lebanese children (10-16 years old) were interviewed using measures of war exposure, mental health symptoms, adaptational outcomes, and Post-Traumatic Stress Disorder (PTSD). The number and type of children's war traumas varied meaningfully in number and type by their age, gender, father's occupational status, and mother's educational level. As predicted, the number of war traumas experienced by a child was positively related to PTSD symptoms; and various types of war traumas were differentially related to PTSD, mental health symptoms, and adaptational outcomes. For example, children who were exposed to multiple war traumas, were bereaved, became victims of violent acts, witnessed violent acts, and/or were exposed to shelling or combat exhibited more PTSD symptoms. Children who were separated from parents reported more depressive symptoms and children who experience bereavement and were not displaced reported more planful behavior. Lastly, children who were separated from parents and who witnessed violent acts reported more prosocial behavior. Implications for program interventions and directions for future research on the effects of war on the psychosocial development of children are explored.


Validation of the Italian version of the Neuropathic Pain Symptom Inventory in peripheral nervous system diseases.
The aim of this study was to validate the Italian version of the Neuropathic Pain Symptom Inventory (NPSI) in patients with neuropathic pain due to peripheral nerve diseases, and also to evaluate the validity of a new NPSI score: a frequency weighted NPSI score (NPSI-FW). First, the original version of the NPSI was translated into Italian. Then the validity and reliability of the Italian NPSI (I-NPSI) were tested in 392 Italian patients consecutively referred to 16 Italian outpatient services for peripheral nerve diseases, by correlating the I-NPSI scores with other pain scales. The repeatability and responsiveness were assessed. A significant correlation between the I-NPSI scores and all the other pain measures was seen. Reproducibility and responsiveness were good. Our study shows the validity of the I-NPSI and demonstrates its reliability for assessing neuropathic pain in patients with peripheral nerve diseases. The I-NPSI scores represent reliable measurements to assess neuropathic symptoms and effectiveness of treatment on them.


An Investigation into the Quality of Life of Cancer Patients in South Africa.
Cancer patients in Africa face unique challenges such as poverty, access to health care and under-resourced health-care systems. Although quality of life (QoL) of cancer patients has been well researched, the perspectives of cancer patients living in Africa are unknown. The objective was to explore what constitutes QoL for cancer patients accessing public health care in South Africa. A qualitative exploratory design was used, and data were gathered by means of in-depth interviews. Purposive sampling selected the participants, and the sample size was determined by saturation (n = 22). The data were analyzed using Patton's method of content analysis. The participants were aged between 20 and 79 years, with an average of 50 years. Most were female and represented seven cultural groups. Four themes that influence QoL arose from the data: psychosocial-, physical-, spiritual and financial factors. QoL remains a complex phenomenon, enhanced and diminished by various individual factors. Poverty was a major issue and influenced the physical aspects of QoL, as the participants had to be strong enough to work and earn a living. Support from family, friends, and church members enhanced QoL, as well as religion and religious practices. Measuring QoL would be the next step to enable nurses to implement measures to improve QoL. Whether existing QoL instruments would be suitable for this patient population is not known and should be investigated before implementation.


Combined array comparative genomic hybridization and tissue microarray analysis suggest PAK1 at 11q13.5-q14 as a critical oncogene target in ovarian carcinoma.
Amplification of chromosomal regions leads to an increase of DNA copy numbers and expression of oncogenes in many human tumors. The identification of tumor-specific oncogene targets has potential diagnostic and therapeutic implications. To identify distinct spectra of oncogenic alterations in ovarian carcinoma, metaphase comparative genomic hybridization (mCGH), array CGH (aCGH), and ovarian tumor tissue microarrays were used in this study. Twenty-six primary ovarian carcinomas and three ovarian carcinoma cell lines were analyzed by mCGH. Frequent chromosomal overrepresentation was observed on 2q (31%), 3q (38%), 5p (38%), 8q (52%), 11q (21%), 12p (21%), 17q (21%), and 20q (52%). The role of oncogenes residing in gained chromosomal loci was determined by aCGH with 59 genetic loci commonly amplified in human tumors. DNA copy number gains were most frequently observed for PIK3CA on 3q (66%), PAK1 on 11q (59%), KRAS2 on 12p (55%), and STK15 on 20q (55%). The 11q13-q14 amplicon, represented by six oncogenes (CCND1, FGF4, FGF3, EMS1, GARP, and PAK1) revealed preferential gene copy number gains of PAK1, which is located at 11q13.5-q14. Amplification and protein expression status of both PAK1 and CCND1 were further examined by fluorescence in situ hybridization and immunohistochemistry using a tissue microarray consisting of 268 primary ovarian tumors. PAK1 copy number gains were observed in 30% of the ovarian carcinomas and PAK1 protein was expressed in 85% of the tumors. PAK1 gains were associated with high grade (P < 0.05). In contrast, CCND1 gene alterations and protein expression were less frequent (10.6% and 25%, respectively), suggesting that the critical oncogene target of amplicon 11q13-14 lies distal to CCND1. This study demonstrates that aCGH facilitates further characterization of oncogene candidates residing in amplicons defined by mCGH.


Pregnancy, abortion, and birth rates among US adolescents--1980, 1985, and 1990.
To analyze pregnancy, abortion, and birth rates among US adolescent girls in 1980, 1985, and 1990. Retrospective analysis of trends in data on pregnancies, abortions, and births. US adolescent girls aged 13 to 19 years. Pregnancy, abortion, and birth rates (with and without adjustment for sexual experience) among teenaged girls aged 15 to 19 years and girls under 15 years. Although pregnancy rates among all teenaged girls 15 to 19 years old remained fairly stable from 1980 to 1985, they increased by 9% during the last half of the decade, totaling 95.9 pregnancies per 1000 teenaged girls 15 to 19 years old by 1990. Because rates of sexual experience increased even faster, pregnancy rates among sexually experienced teens aged 15 to 19 actually declined between 1980 and 1990 by approximately 8%. Abortion rates among these teens remained stable during the 1980s, with 35.8 and 36.0 abortions per 1000 in 1980 and 1990, respectively. As with overall pregnancy rates, abortion rates among these sexually experienced teenaged girls declined during the 1980s. Between 1980 and 1985, birth rates among teenaged girls aged 15 to 19 years declined by 4%, but they increased by 18% during the latter half of the decade, totaling 59.9 births per 1000 in 1990. Among these sexually experienced teenagers, birth rates also declined between 1980 and 1985 and then increased over the next 5 years. In 1990, pregnancies and abortions among girls younger than 15 years accounted for only 3% of all adolescent pregnancies and abortions. However, the number of births among these younger adolescents increased by 15% over the decade. In that age group, trends in pregnancy, abortion and birth rates over the decade were similar to those for older teens. However, during the late 1980s, the abortion rate declined and the pregnancy rate remained stable, resulting in a 26% increase in the birth rate. Despite efforts to reduce adolescent pregnancy in the United States, pregnancy and birth rates for that group continue to be the highest among developed countries. Considering that 95% of adolescent pregnancies are unintended, increased efforts to prevent these pregnancies are warranted.


Tumescent local anesthesia for excision and flap procedures in treatment of pilonidal disease.
We describe our experience with tumescent local anesthesia for excision and flap procedures in the treatment of pilonidal disease. Forty consecutive patients with symptomatic pilonidal disease were treated. Tumescent solution was prepared by mixing one volume of a lidocaine (20 mg/ml) and adrenaline (0.0125 mg/ml) combination with 9 volumes of lactated Ringer's solution. The final solution was infiltrated gradually and liberally, first subcutaneously and then into the skin. The end point was swollen and firm skin. After excision, the area was filled with an advancement flap or a rotational flap. The study group consisted of 37 men (92.5%) and 3 women (7.5%); mean age, 27 (range, 18-61) years. The amount of injected solution ranged from 60 mL to 140 mL (mean, 100 mL; dose interval, 2-4 mg/kg). No additional anesthesia or sedation was needed, and no complications were observed during the procedure. Half of the patients were discharged on the day of the operation, half on the next day. Follow-up ranged from 13 to 37 months. Patients returned to daily activities after a mean of 10.5 (range, 2-30) days. Primary healing occurred uneventfully in 28 patients (70%); wound healing complications occurred in 12 (30%). No flap necrosis was observed. Thirty-seven (92.5%) patients reported satisfaction with the procedure. Recurrence was observed in 3 patients (7.5%). Excision and flap procedures for sacrococcygeal pilonidal disease can be performed under tumescent local infiltration anesthesia with acceptable results, without requiring regional or general anesthesia.


Absence of OKT4 antigen epitope: report of a case in a Brazilian population.
During the study of 162 normal subjects we found an 8 month-old black girl without previous illnesses, especially infections, whose lymphocytes did not react with OKT4 monoclonal antibody. However the CD4 subset was present, since the number of lymphocytes which reacted with Leu 3a and T4 was normal. In addition the percentage of Fc mu+ lymphocytes, K cell activity and PHA response were normal. The proportion of the other lymphocyte subsets did not differ from normal controls. Four first degree relatives of this child had a normal proportion of OKT4+ lymphocytes. The absence of OKT4 epitope was previously reported as a rare genetic variant which was detected only in Japanese and in subjects of black African ancestry.


Heuristic map of myotomal innervation in humans using direct intraoperative nerve root stimulation.
Considerable overlap exists in nerve root innervation of various muscles. Knowledge of myotomal innervation is essential for the interpretation of neurological examination findings and neurosurgical decision-making. Previous studies relied on cadaveric dissections, animal studies, and cases with anomalous anatomy. This study investigates the myotomal innervation patterns of cervical and lumbar nerve roots through in vivo stimulation during surgeries for spinal decompression. Patients undergoing cervical and lumbar surgeries in which nerve roots were exposed in the normal course of surgery were included in the study. Electromyography electrodes were placed in the muscle groups that are generally accepted to be innervated by the roots under study. These locations included levels above and below the spinal levels undergoing decompression. After decompression, a unipolar neural stimulator probe was placed directly on the nerve root sleeve and constant current stimulation in increments of 0.1 mA was performed. Current was raised until at least a 100 μV amplitude-triggered electromyographic response was noted in 1 or more muscles. All muscles that responded were recorded. A total of 2295 nerve root locations in 129 patients (mean age 57 ± 15 years, 47 female [36%]) were stimulated, and 1589 stimulations met quality criteria and were analyzed. Four hundred ninety-five stimulations were performed on roots contributing to the cervical and brachial plexus from C-3 to T-1 (31.2%), and 1094 (68.8%) were roots in the lumbosacral plexus between L-1 and S-2. The authors were able to construct a statistical map of the contributions of each cervical and lumbosacral nerve root for the set of muscle groups monitored in the protocol. In many cases the range of muscles innervated by a specific root was broader than previously described in textbooks. This is the largest data set of direct intraoperative nerve root stimulations during decompressive surgery, demonstrating the relative contribution of root-level motor input to various muscle groups. Compared with classic neuroanatomy, a significant number of roots innervate a broader range of muscles than expected, which may account for the variability of presentation between patients with identical number and location of compressed roots.


Binding and internalization of transforming growth factor-beta 1 by human hepatoma cells: evidence for receptor recycling.
Cellular processing of 125I-labeled transforming growth factor-beta 1 was investigated in the human hepatoma cell lines Hep G2 and Hep 3B. Binding of 125I-transforming growth factor-beta 1 to cell surface receptors was specific, saturable and calcium-independent. Both cell lines exhibited a single class of high-affinity (Kd = 2.2 x 10(-10) mol/L) binding sites (4.5 x 10(3) for the Hep G2 cell; 1.5 x 10(3) for the Hep 3B cell) for both human and porcine transforming growth factor-beta 1. Binding was temperature dependent, time dependent and pH dependent. Cell-bound 125I-transforming growth factor-beta 1 was removed by brief exposure to acidic medium (pH less than 4) but was converted into an acid-resistant state rapidly after shifting the cells to 37 degrees C. Spontaneous dissociation of bound ligand over a 6 hr period at 4 degrees C was less than 10%. Disuccinimidyl suberate was used to covalently label 125I-transforming growth factor-beta 1 to cell-surface binding sites. Labeling of the ligand/receptor complexes was inhibited by unlabeled transforming growth factor-beta 1 but was unaffected by other growth factors. The radiolabeled complexes showed approximate molecular weights of 280,000, 85,000 and 65,000 when run on reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cell-bound 125I-transforming growth factor-beta 1 was internalized and degraded at 37 degrees C, and the products were released into the medium as trichloroacetic acid-nonprecipitable radioactivity. The lysosomotropic base chloroquine and the carboxylic ionphore monensin inhibited degradation and release of 125I-labeled products from the cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Plasma-Enhanced Atomic Layer Deposition of Nanostructured Gold Near Room Temperature.
A plasma-enhanced atomic layer deposition (PE-ALD) process to deposit metallic gold is reported, using the previously reported Me3Au(PMe3) precursor with H2 plasma as the reactant. The process has a deposition window from 50 to 120 °C with a growth rate of 0.030 ± 0.002 nm per cycle on gold seed layers, and it shows saturating behavior for both the precursor and reactant exposure. X-ray photoelectron spectroscopy measurements show that the gold films deposited at 120 °C are of higher purity than the previously reported ones (<1 at. % carbon and oxygen impurities and <0.1 at. % phosphorous). A low resistivity value was obtained (5.9 ± 0.3 μΩ cm), and X-ray diffraction measurements confirm that films deposited at 50 and 120 °C are polycrystalline. The process forms gold nanoparticles on oxide surfaces, which coalesce into wormlike nanostructures during deposition. Nanostructures grown at 120 °C are evaluated as substrates for free-space surface-enhanced Raman spectroscopy (SERS) and exhibit an excellent enhancement factor that is without optimization, only one order of magnitude weaker than state-of-the-art gold nanodome substrates. The reported gold PE-ALD process therefore offers a deposition method to create SERS substrates that are template-free and does not require lithography. Using this process, it is possible to deposit nanostructured gold layers at low temperatures on complex three-dimensional (3D) substrates, opening up opportunities for the application of gold ALD in flexible electronics, heterogeneous catalysis, or the preparation of 3D SERS substrates.


Decrease of RA-related orthopaedic surgery of the upper limbs between 1998 and 2004: data from 54,579 Swedish RA inpatients.
To describe the overall use and temporal trends in orthopaedic upper limb surgery associated with RA on a nation wide basis in Sweden between 1998 and 2004. Data for all inpatient visits during 1998-2004 for patients older than 18 yrs with RA-related diagnoses were extracted from the Swedish National Hospital Discharge Registry (SNHDR). The SNHDR prospectively collects data on all hospital admissions in Sweden according to the International Classification of Diseases (ICD). Data were analysed with respect to orthopaedic surgery of the hand, elbow and shoulder. During the study period, 54,579 individual RA patients were admitted to a Swedish hospital and 9% of these underwent RA-related surgery of the upper limbs. The RA patient cohort underwent a total of 8251 RA-related upper limb surgical procedures. The hand (77%) was most frequently operated on, followed by the shoulder (13%) and the elbow (10%). There was a statistically significant decrease of 31% for all admissions associated with RA-related upper limb surgery during 1998-2004 (P = 0.001). Some 10% of all RA-related upper limb surgery was due to total joint arthroplasties (TJAs), mostly for the elbow (59%). During 1998-2004, all TJAs, elbow-TJAs and shoulder-TJAs had a stable occurrence. In contrast, the overall numbers of hand-TJAs significantly increased (P = 0.009). Rates of RA-related upper limb surgery decreased and TJAs had a stable occurrence in Sweden during 1998-2004. The findings of this study may reflect trends in disease management and health outcomes of RA patients in Sweden.


Lightning-induced high temperature and pressure microstructures in surface and subsurface fulgurites.
Cloud-to-ground lightning causes both high-temperature and high-pressure metamorphism of rocks, forming rock fulgurite. We demonstrate that a range of microstructural features indicative of high temperatures and pressures can form in fulgurites at the surface and in fractures up to several meters below the surface. In comparison to a granite reference sample collected from a borehole at a depth of 138 m, microstructures in both the surface and fracture fulgurite are characterized by: (i) the presence of glass, (ii) a phase transformation in K-feldspar with the presence of exsolution lamellae of plagioclase, and (iii) high residual stresses up to 1.5 GPa. Since this is the first time that fracture-related fulgurite has been described, we also carried out a 1-D numerical model to investigate the processes by which these can form. The model shows that the electric current density in fractures up to 40 m from the landing point can be as high as that on the surface, providing an explanation for the occurrence of fracture-related fulgurites. Our work broadens the near-surface environments in which rock fulgurite has been reported, and provides a detailed description of microstructures that can be compared to those formed during other types of extreme metamorphic events.


Diffusion-weighted imaging of prostate cancer.
The purpose of this study was to assess whether T2-weighted (T2W) imaging with diffusion-weighted (DW) imaging could improve prostate cancer detection as compared with T2W imaging alone. The subjects consisted of 37 patients with prostate cancer and 23 without cancer undergoing magnetic resonance (MR) imaging. Using a 1.5-T superconducting magnet, all patients underwent T2W and DW imaging with parallel imaging. Images were independently reviewed by 3 readers to determine the detectability of prostate cancer. The detectability of T2W imaging without and with DW imaging was assessed by means of receiver operating characteristic analysis. Mean areas under the receiver operating characteristic curve for T2W imaging alone and for T2W imaging with DW imaging were 0.87 and 0.93, respectively. The receiver operating characteristic analysis showed that the addition of DW imaging to conventional T2W imaging significantly improved tumor detection (P = 0.0468) compared with T2W imaging alone. The addition of DW imaging to conventional T2W imaging provides better detection of prostate cancer.


[Laser-induced covalent cross-links in DNA].
Formation of cross-links and local denaturated sites in double-stranded DNA of bacteriophage CD under picosecond laser UV irradiation was investigated by fluorescent method. It is shown that passing from low-intensity UV irradiation with mercury lamp to high-intensity laser UV irradiation quantum, yield of cross-links formation increases by an order.


Extracellular signal-regulated kinase activation is required for up-regulation of vascular endothelial growth factor by serum starvation in human colon carcinoma cells.
Vascular endothelial growth factor (VEGF) is a potent angiogenic factor important for colon cancer neovascularization. In previous studies, serum starvation led to induction of VEGF in human colon carcinoma cells. We investigated the possible participation of mitogen-activated protein kinases in serum starvation induction of VEGF in the HT29 human colon carcinoma cell line. The extracellular signal-regulated kinases (Erks) 1 and 2 were activated after 3-6 h of serum starvation. Using transient transfection of VEGF promoter-reporter constructs, serum starvation led to an increase in VEGF promoter activity. An inhibitor of phosphorylation of Erk-1/2 blocked the increase of VEGF expression and promoter activity induced by serum starvation. Serum starvation activates several mitogen-activated protein kinases, but activation of Erk-1/2 is critical for the up-regulation of VEGF mRNA in colon carcinoma cells.


Amounts and types of fatty acids in meals affect the pattern of retinoids secreted in human chylomicrons after a high-dose preformed vitamin A intake.
High doses of preformed vitamin A are commonly used to correct vitamin A deficiency. Newly absorbed vitamin A is secreted mainly as retinyl esters in chylomicrons. The effect of changing types and amounts of fatty acids on fatty acid composition of chylomicron retinoid esters when a high dose of vitamin A is ingested have not been studied previously. In the present study, 10 healthy young men ingested, in a random order, mixed meals containing 15,000 retinol equivalents (RE) of vitamin A (as retinyl palmitate) and either no fat or 40 g of fat provided as butter, olive oil, or sunflower oil. Fasting and postprandial blood samples were obtained for 7 hours after meals. Free retinol and the main retinyl esters (retinyl palmitate/oleate, stearate, and linoleate) were measured in chylomicrons by high-performance liquid chromatography (HPLC). Chylomicron retinyl palmitate/oleate and retinyl stearate concentrations significantly increased after intake of the 4 test meals. Conversely, chylomicron retinyl linoleate and chylomicron free retinol significantly increased only after the sunflower and the fat-free meals, respectively. The main retinoid secreted in chylomicrons after the intake of the fat-rich meals was retinyl palmitate/oleate, accounting for 63% to 79% of total RE, but it was free retinol after the fat-free meal (51% of total RE). Thus, the retinoid pattern secreted in chylomicrons after the intake of a high dose of preformed vitamin A depends on type and amounts of fatty acids ingested. To explain this result we suggest that the esterification process of retinol in the enterocyte by lecithin:retinol acyltransferase can be overwhelmed by a high load of vitamin A. Consequently, a significant proportion of the retinol is esterified by acyl coenzyme A:retinol acyltransferase (ARAT) with ingested fatty acids, explaining the appearance of retinyl linoleate in chylomicrons after the sunflower oil meal. If a high dose of preformed vitamin A is ingested with a fat-free meal, a significant proportion of retinol is not esterified, owing to the lack of fatty acids for ARAT, which explains the appearance of free retinol in chylomicrons.


Cardiac Point-of-Care Ultrasound (P.O.C.U.S.) Utilization for Hospitalists in the Assessment of Patients with Cardiac Complaints: An Educational Overview.
The use of cardiac point-of-care ultrasound (P.O.C.U.S.) is underutilized in the field of internal medicine for the assessment of patients with cardiac complaints. Numerous studies in emergency medicine, anesthesia, and critical care have demonstrated the successful application of cardiac P.O.C.U.S. in resident and attending physicians with limited prior exposure. This article review overviews the practical implementation of cardiac P.O.C.U.S. for hospitalists by discussing proper technique and assessment for common pathology seen in the medical ward setting. We describe how to assess for left ventricular (LV) systolic function, right ventricular (RV) systolic function, suspected acute coronary syndrome (ACS), post-myocardial infarction (MI) complications, suspected pulmonary embolus, and assessment of intravascular volume status. In each section, we overview the pertinent literature to show how cardiac P.O.C.U.S. has been used to directly impact patient care.


Nociceptin/orphanin FQ regulates neuroendocrine function of the limbic-hypothalamic-pituitary-adrenal axis.
We examined the effects of the neuropeptide nociceptin/orphanin FQ on activity of the limbic-hypothalamic-pituitary-adrenal axis (also known as the stress axis) in rats. This axis regulates important metabolic functions, and initiates critical neuroendocrine responses that cope with environmental threats and challenges to homeostatic functioning. Disregulation of the limbic-hypothalamic-pituitary-adrenal axis is associated with impaired physical and psychological health. In the present experiments, rats were treated with intracerebroventricular injections of nociceptin/orphanin FQ in the presence or absence of acute stressors. Plasma adrenocorticotrophic hormone and corticosterone concentrations were assayed 15 or 30min after injections. In the rats that were not exposed to stress, nociceptin/orphanin FQ produced dose-orderly elevations of circulating adrenocorticotrophic hormone and corticosterone concentrations. These effects were also found after administration of the nociceptin/orphanin FQ analogues, des-Phe orphanin FQ and [Phe(1)psi(CH(2)-NH)Gly(2)]nociceptin((1-13))NH(2). In rats that were exposed to the mild stress of a novel environment, nociceptin/orphanin FQ administration enhanced the stress-induced elevations of plasma adrenocorticotrophic hormone concentrations and prolonged the stress-induced elevations of plasma corticosterone concentrations. In rats that were exposed to restraint stress, nociceptin/orphanin FQ administration did not augment the stress-induced elevations in plasma hormones, perhaps because of a ceiling effect. We conclude that administration of nociceptin/orphanin FQ activates neuroendocrine activity of the limbic-hypothalamic-pituitary-adrenal axis even in the absence of a stressor, and may delay the shutdown of these physiological responses after exposure to acute mild stress. In light of the known functions of this axis, it appears that nociceptin/orphanin FQ participates in the regulation of important metabolic functions, and may be implicated in physiological responses to stress. This interaction between nociceptin/orphanin FQ and the limbic-hypothalamic-pituitary-adrenal axis implicates nociceptin/orphanin FQ in important aspects of physiological and psychological well-being.


Enterococci in insects.
Enterococci were obtained from 213 of 403 insects cultured during a 14-month period, in numbers from 10(3) to 3 x 10(7)/g of insect. Insects were taken only from nonurban, wild, and cultivated fields and woods. In species of insects carrying them, enterococci were not always present in every individual cultured, and often more than one species of enterococcus occurred within a species. Enterococci were obtained from certain insects taken in the field during the dormant season, suggesting their role as overwintering agents. They were generally present in species feeding on nectar, succulent plant parts, and on and ir forest litter, but not from insects feeding on less succulent leaves and stems. Streptococcus faecalis was recovered from 32%, Streptococcus faecium from 22.4%, and Streptococcus faecium var. casseliflavus from 43.5% of members of the 37 taxa of insects. S. faecalis and S. faecium var. casseliflavus exhibit a high percent of conformity to the properties published for them. The heterogeneity in properties of S. faecium is similar to that found for the species taken from plants. Many fail to grow in broth at 45 C or in broth containing 6.5% NaCl; 50% of the cultures ferment both melezitose and melibiose, and a few ferment neither sugar. The remainder ferment melibiose only. Failure to reduce methylene blue in milk by S. faecalis and S. faecium is correlated with the inability to ferment lactose. More than 93% of the cultures of S. faecalis digest casein in milk from the top downward, following the production of a soft, flowing curd. Because this property is not characteristic of S. faecalis taken from humans, the reaction in litmus milk is suggested as a means of differentiation between cultures of remote and innocent origin in nature and recent, human pollution.


Impact of Dietary Fats on Brain Functions.
Adequate dietary intake and nutritional status have important effects on brain functions and on brain health. Energy intake and specific nutrients excess or deficiency from diet differently affect cognitive processes, emotions, behaviour, neuroendocrine functions and synaptic plasticity with possible protective or detrimental effects on neuronal physiology. Lipids, in particular, play structural and functional roles in neurons. Here the importance of dietary fats and the need to understand the brain mechanisms activated by peripheral and central metabolic sensors. Thus, the manipulation of lifestyle factors such as dietary interventions may represent a successful therapeutic approach to maintain and preserve brain health along lifespan. This review aims at summarizing the impact of dietary fats on brain functions. Starting from fat consumption, nutrient sensing and food-related reward, the impact of gut-brain communications will be discussed in brain health and disease. A specific focus will be on the impact of fats on the molecular pathways within the hypothalamus involved in the control of reproduction via the expression and the release of Gonadotropin-Releasing Hormone. Lastly, the effects of specific lipid classes such as polyunsaturated fatty acids and of the "fattest" of all diets, commonly known as "ketogenic diets", on brain functions will also be discussed. Despite the knowledge of the molecular mechanisms is still a work in progress, the clinical relevance of the manipulation of dietary fats is well acknowledged and such manipulations are in fact currently in use for the treatment of brain diseases.


Trialling nutrient recommendations for slow lorises (Nycticebus spp.) based on wild feeding ecology.
Slow loris (Nycticebus spp.) captive diets have been based on routine and anecdotes rather than scientific fact. The growing body of evidence contradicts the high fruit diet supported by such anecdotes. Non-human primate nutrient requirements are grouped into new (based on the common marmoset Callithrix jacchus) or old world (based on rhesus macaques Macaca mulatta) primates. Slow lorises are known to suffer from many health ailments in captivity such as dental disease, obesity, wasting and kidney issues all of which have been linked to diet. This study aimed to estimate nutrient intake from free-ranging slow lorises and to determine whether this intake can be used as nutrient recommendations. We collected data of nutrient intake, food passage rate and digestibility of captive slow lorises on three diet treatments 1: current captive type diet which is mostly fruits, 2: wild-type diet made only of food items from their natural diet, 3: new diet made to reflect wild slow loris nutrient intake. In order to validate our nutrient recommendations, diets 2 and 3 would have to be significantly different to Diet 1 in terms of nutrients, but not different from each other. Captive diets were significantly higher in soluble carbohydrates and lower in minerals and fibre fractions than both diets 2 and 3. Diets 2 and 3 led to a significantly increased food passage time and to more effective fibre and calcium digestion compared to Diet 1. We also observed obese individuals lost weight and underweight individuals gained weight. Our nutrient recommendations have been validated by our trials, and new or old world monkey nutrient recommendations are not consistent with our results. Diets should be high in protein and fibre and low in soluble carbohydrates and fats.


Renal xenobiotic transporters are differentially expressed in mice following cisplatin treatment.
The goal of this study was to identify alterations in mRNA and protein expression of various xenobiotic transport proteins in mouse kidney during cisplatin-induced acute renal failure. For this purpose, male C57BL/6J mice received a single dose of cisplatin (18 mg/kg, i.p.) or vehicle. Four days later, tissues were collected for assessment of plasma BUN, histopathological analysis of renal lesions, and mRNA and Western blot analysis of renal transporters including organic anion and cation transporters (Oat, Oct), organic anion transporting polypeptides (Oatp), multidrug resistance-associated proteins (Mrp), multidrug resistance proteins (Mdr), breast cancer resistance protein (Bcrp) and multidrug and toxin extrusion proteins (Mate). Cisplatin treatment caused necrosis of renal proximal tubules along with elevated plasma BUN and renal kidney injury molecule-1 mRNA expression. Cisplatin-induced renal injury increased mRNA and protein levels of the efflux transporters Mrp2, Mrp4, Mrp5, Mdr1a and Mdr1b. Uptake transporters Oatp2a1 and Oatp2b1 mRNA were also up-regulated following cisplatin. By contrast, expression of Oat1, Oat2, Oct2 and Oatp1a1 mRNA was reduced in cisplatin-treated mice. Expression of several uptake and efflux transporters was unchanged in cisplatin-treated mice. Apical staining of Mrp2 and Mrp4 proteins was enhanced in proximal tubules from cisplatin-treated mice. Collectively, these expression patterns suggest coordinated regulation of uptake and efflux pathways during cisplatin-induced renal injury. Reduced expression of basolateral and apical uptake transporters along with enhanced transcription of export transporters likely represents an adaptation to lower intracellular accumulation of chemicals, prevent their reabsorption and enhance urinary clearance.


Intentionality, Morality, and the Incest Taboo in Madagascar.
In a recent article (Astuti and Bloch, 2015), cognitive anthropologists Astuti and Bloch claim that the Malagasy are ambivalent as to whether considerations of intentionality are relevant to moral judgments concerning incest and its presumed catastrophic consequences: when making moral judgments about those who commit incest, the Malagasy take into account whether the incest is intentional or not, but, when making moral judgments relating to incest's catastrophic consequences, they do not take intentionality into account. Astuti and Bloch explain the irrelevance of intentionality in terms of incest entailing such a fundamental attack on the transcendental social order that the Malagasy become dumbfounded and leave aside considerations of intentionality. Finally, they claim that a similar dumbfound reaction is what is involved in the moral dumbfounding concerning incest that social psychologist Jonathan Haidt has found in the US. In this article, we argue that (i) Astuti and Bloch are unclear about many aspects of their claims (in particular, about the moral judgments at stake), (ii) they do not provide sufficient evidence that considerations of intentionality are deemed irrelevant to moral judgments relating to incest's presumed catastrophic consequences (and hence for the claim that the Malagasy are ambivalent), (iii) their hypothesis that conceiving of incest as an attack on the transcendental social renders considerations of intentionality irrelevant lacks coherence, and (iv) the extension of their explanatory account to the moral dumfounding of American students in Haidt's well-known scenario of intentional incest is unwarranted.


[Spread of Yersinia among patients with acute intestinal infections].
During 1979 a large-scale study of fecal samples obtained from patients with acute intestinal infections was carried out with the aim of establishing the presence of Yersinia. As a result, 9 Y. enterocolitica strains and 7 Y. pseudotuberculosis strains were isolated. Y. enterocolitica were classified with serovars O8, O3, and O5B and with biovars 2, 3, and 4; Y. pseudotuberculosis were classified with serovars 1 and 3. The isolation of Yersinia facilitated the identification of the etiological structure of acute intestinal infections.


Transient stress lymphocytosis: an immunophenotypic characterization of the most common cause of newly identified adult lymphocytosis in a tertiary hospital.
We prospectively evaluated 52 consecutive cases of newly identified absolute lymphocytosis to determine the hematologic and immunophenotypic features of transient stress lymphocytosis (TSL). The lymphocytosis in all cases was associated with an acute stressful event and ranged from 4,000 to 10,400/microL (4.0-10.4 x 10(9)/L). Compared with healthy individuals, patients with TSL showed an increase in the total WBC, absolute lymphocyte (ALC), absolute neutrophil (ANC), and platelet counts with no difference in hemoglobin levels. Immunophenotypic analyses of 38 cases revealed increases in absolute numbers of T B, and natural killer cells. Both CD4+ and CD8+ T cells were increased, predominantly accountedfor by an increase in memory cell subsets, with no change in gamma/delta T cells. Follow-up studies showed a significant reduction in the ALC with a concurrent increase in the ANC and reduction in hemoglobin values. The reduction in lymphocytes at resolution was accompanied by reduction in all broad Iymphocyte subsets. However, naive and memory subsets showed different patterns of alteration within the CD4+ and CD8+ populations, suggesting that acute stress differentially affects the in vivo distribution of these subsets.


A promoter element active in run-off transcription controls the expression of two cistrons of nad and rps genes in Nicotiana sylvestris mitochondria.
The expression of two mitochondrial gene clusters (orf87-nad3-nad1/A and orf87-nad3-rps12) was studied in Nicotiana sylvestris. 5' and 3' termini of transcripts were mapped by primer extension and nuclease S1 protection. Processing and transcription initiation sites were differentiated by in vitro phosphorylation and capping experiments. A transcription initiation site, present in both gene clusters, was found 213 nucleotides upstream of orf87. This promoter element matches the consensus motif for dicotyledonous mitochondrial promoters and initiates run-off transcription in a pea mitochondrial purified protein fraction. Processing sites were identified 5' of nad3, nad1/A and rps12 respectively. These results suggest that (i) the expression of the two cistrons is only controlled by one duplicated promoter element, and (ii) multiple processing events are required to produce monocistronic nad3, nad1/A and rps12 transcripts.


Effectiveness of self-management applications in improving clinical health outcomes and adherence among diabetic individuals in low and middle-income countries: a systematic review.
A variety of mobile health (mHealth) applications are available to monitor an individual's health or lifestyle to make it convenient to access healthcare facilities at home. The usability of mHealth applications in controlling HbA1c (estimated average blood glucose) levels is unclear despite their increasing use. The burden of type 2 diabetes mellitus (T2DM) is high in low and middle-income countries (LMICs), with the highest burden in the Indian population. Our objective is to identify the effectiveness of mHealth applications in managing blood glucose levels of individuals with T2DM and to assess the impact of using mHealth applications in managing T2DM concerning health-promoting behaviour among the LMICs in the context of India. The electronic databases included for search are PubMed, Ovid Medline, EBSCO, CINAHL, Scopus, Web of Science and the Cochrane Central Register of Controlled Trials; additional sources of the search will be grey literature available on diabetes management websites and reference lists of included studies. Studies published in the English language in indexed and peer-reviewed sources will be considered. Studies reporting the effectiveness of mobile applications in the management of T2D in LMICs will be eligible for inclusion. The Population-Intervention-Comparison-Outcomes framework and the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement 2021 will be used for reporting. Data analysis will be carried out using narrative synthesis, and a meta-analysis may be conducted if we come across homogenous data for the outcome. As this study is a systematic review, we will not be recruiting any participants for the study and hence will not require ethical approval. The study summary will be disseminated at a conference. CRD42021245517.


Comparison of random amplified polymorphic DNA markers and morphological characters in identification of homokaryon isolates of white button mushroom (Agaricus bisporus).
The secondarily homothallic life cycle of the white button mushroom that results in scarcity of uninucleate basidiospores (homokaryons) in its progeny, is the most important impediment for genetic improvement of the commercial strains. Identification of homokaryons for breeding programs of Agaricus bisporus (button mushroom) is, therefore, crucial. Verifying homokaryons through fruiting trial is time consuming and unreliable. In this study, ability of RAPD markers, compared to morphological characters for identification of homokaryon isolates, was investigated. Based on morphological characters, 42 isolates were screened and exposed to RAPD markers. The results showed that RAPD markers could discriminate homokaryons from heterokaryons, based on number of bands generated. The numbers of band in homokaryons were significantly less than those of heterokaryons. Results also showed that cluster analysis, based on average of band number generated, could separate homokaryon from heterokaryon isolates. It is suggested that RAPDs could be used to identify hyomokaryons from heterokaryons for breeding program of A. bisporus.


Individual fates of mesenchymal stem cells in vitro.
In vitro cultivated stem cell populations are in general heterogeneous with respect to their expression of differentiation markers. In hematopoietic progenitor populations, this heterogeneity has been shown to regenerate within days from isolated subpopulations defined by high or low marker expression. This kind of plasticity has been suggested to be a fundamental feature of mesenchymal stem cells (MSCs) as well. Here, we study MSC plasticity on the level of individual cells applying a multi-scale computer model that is based on the concept of noise-driven stem cell differentiation. By simulation studies, we provide detailed insight into the kinetics of MSC organisation. Monitoring the fates of individual cells in high and low oxygen culture, we calculated the average transition times of individual cells into stem cell and differentiated states. We predict that at low oxygen the heterogeneity of a MSC population with respect to differentiation regenerates from any selected subpopulation in about two days. At high oxygen, regeneration becomes substantially slowed down. Simulation results on the composition of the functional stem cell pool of MSC populations suggest that most of the cells that constitute this pool originate from more differentiated cells. Individual cell-based models are well-suited to provide quantitative predictions on essential features of the spatio-temporal organisation of MSC in vitro. Our predictions on MSC plasticity and its dependence on the environment motivate a number of in vitro experiments for validation. They may contribute to a better understanding of MSC organisation in vitro, including features of clonal expansion, environmental adaptation and stem cell ageing.


Clinical effects of applying a tourniquet in total knee arthroplasty on blood loss.
Tourniquets used during total knee arthroplasty may lead to many complications. The aim of this study was to determine perioperative blood loss and its clinical relevance in total knee replacement surgery after applying a tourniquet. From June 2009 to October 2009, 60 consecutive patients who underwent routine total knee arthroplasty were randomly divided into two groups and were treated with or without a tourniquet (30 patients/group). There were no significant differences in patient baseline characteristics between the two groups. We compared the two groups of patients in terms of intra- and postoperative bleeding, invisible or visible bleeding, and total blood loss. None of the patients showed poor wound healing, lower extremity deep venous thrombosis or other complications. The amount of blood loss during surgery was lower in the tourniquet group than in the control group (P < 0.01). However, postoperative visible bleeding (P < 0.05) and occult bleeding (P < 0.05) were significantly greater in the tourniquet group than in the control group. There was no significant difference in the total amount of blood loss between the two groups (P > 0.05). Tourniquet can reduce bleeding during total knee replacement surgery, but is associated with greater visible and invisible blood loss.


IMFLer: A Web Application for Interactive Metabolic Flux Analysis and Visualization.
Increasing genome-wide data in biological sciences and medicine has contributed to the development of a variety of visualization tools. Several automatic, semiautomatic, and manual visualization tools have already been developed. Some even have integrated flux balance analysis (FBA), but in most cases, it depends on separately installed third party software that is proprietary and does not allow customization of its functionality and has many restrictions for easy data distribution and analysis. In this study, we present an interactive metabolic flux analyzer and visualizer (IMFLer)-a static single-page web application that enables the reading and management of metabolic model layout maps, as well as immediate visualization of results from both FBA and flux variability analysis (FVA). IMFLer uses the Escher Builder tool to load, show, edit, and save metabolic pathway maps. This makes IMFLer an attractive and easily applicable tool with a user-friendly interface. Moreover, it allows to faster interpret results from FBA and FVA and improves data interoperability by using a standardized file format for the genome-scale metabolic model. IMFLer is a fully open-source tool that enables the rapid visualization and interpretation of the results of FBA and FVA with no time setup and no programming skills required, available at https://lv-csbg.github.io/IMFLer/.


MAIT cells altered phenotype and cytotoxicity in lupus patients are linked to renal disease severity and outcome.
Systemic lupus erythematosus (SLE) is an autoimmune disease in which circulating immune complexes can cause different types of glomerulonephritis, according to immune deposits and to the type of glomerular cell injury. Proliferative lesions represent the most severe form of lupus nephritis (LN) and often lead to kidney failure and death. Mucosal-associated invariant T (MAIT) cells are a subset of innate-like T cells that recognize microbial-derived ligands from the riboflavin synthesis pathway. Although abundant in peripheral blood, MAIT cells are enriched in mucosal and inflamed tissues. While previous studies have reported concordant results concerning lower MAIT cell frequencies in the blood of SLE patients, no information is known about MAIT cell function and LN severity and outcome. In the current study, we analyzed the baseline phenotype and function of peripheral blood MAIT cells by flow cytometry in 26 patients with LN and in a control group of 16 healthy individuals. We observe that MAIT cell frequencies are markedly reduced in blood of LN patients. MAIT cells from patients have an altered phenotype in terms of migration, proliferation and differentiation markers, notably in most severe forms of LN. Frequencies of PMA/ionomycin stimulated MAIT cells secreting effector molecules, such as proinflammatory IL-17 and cytotoxic protein granzyme B, are higher in LN patients. Patients undergoing a complete renal remission after immunosuppressive therapy had higher MAIT cell frequency, lower expression of proliferation marker Ki-67 and granzyme B (GzB) at inclusion. Remarkably, GzB production defines a predictive model for complete remission. We report here that blood MAIT cells display proinflammatory and cytotoxic function in severe lupus nephritis which may play a pathogenesis role, but without association with systemic lupus activity. Finally, low cytotoxic profile of MAIT cells may represent a promising prognostic factor of lupus nephritis remission one year after induction therapy.


Phylogeny of European bat Lyssavirus 1 in Eptesicus isabellinus bats, Spain.
To better understand the epidemiology of European bat lyssavirus 1 (EBLV-1) in Europe, we phylogenetically characterized Lyssavirus from Eptesicus isabellinus bats in Spain. An independent cluster of EBLV-1 possibly resulted from geographic isolation and association with a different reservoir from other European strains. EBLV-1 phylogeny is complex and probably associated with host evolutionary history.


Accuracy of incidental visual coronary artery calcium assessment compared with dedicated coronary artery calcium scoring.
CAC can be detected on routine chest computed tomography (CT) scans and may contribute to CVD risk estimation, but the accuracy of visual CAC scoring may be affected by the specialty of the interpreting radiologist and/or the use of contrast. The accuracy of visual CAC estimation on non-gated CT scans was evaluated at UT Southwestern Medical Center (UTSW) and Parkland Health and Hospital System (PHHS). All adults who underwent CAC scanning and a non-gated CT scan within 6 months were identified and the scores from the two CTs were compared overall and stratified by type of reader and whether contrast was used. Visual CAC categories of none, small, moderate, and large were compared to CAC = 0, 1-99, 100-399, and ≥400, respectively. From 2016 to 2021, 934 patients (mean age 60 ± 12 y, 43% male, 61% White, 34% Black, 24% Hispanic, 54% from PHHS) had both CT scans. Of these, 441 (47%) had no CAC, 278 (30%) small, 147 (16%) moderate, and 66 (7%) large CAC on non-gated CT. Visual CAC estimates were highly correlated with CAC scores (Kendalls tau-b = 0.76, p < 0.0001). Among those with no visual CAC, 76% had CAC = 0 (72% of contrast-enhanced vs 85% of non-contrast scans, 88% of scans interpreted by CT radiologist vs 78% of those interpreted by other radiologist). In those with moderate-to-large visual CAC, 99% had CAC >0 and 88% had CAC ≥100, including 89% of those with contrast, 90% of those without contrast, 80% of those read by a CT radiologist, and 88% of those read by a non-CT radiologist. Visual CAC estimates on non-gated CT scans are concordant with Agatston score categories from cardiac CT scans. A lack of visual CAC on non-gated CT scans may not be sufficient to "de-risk" patients, particularly for contrast-enhanced scans and those read by non-CT radiologists. However, the presence of moderate-to-large CAC, including on contrasted scans and regardless of radiologist type, is highly predictive of CAC and may be used to identify high-risk patients for prevention interventions.


Slowed motor conduction in lumbosacral nerve roots in cauda equina lesions: a new diagnostic technique.
New techniques have been developed for the electrophysiological assessment of patients with suspected cauda equina lesions using transcutaneous spinal stimulation (500-1500 V: time constant 50 microseconds) to measure motor latencies to the external and sphincter and puborectalis muscles from L1 and L4 vertebral levels. These latencies represent motor conduction in the S3 and S4 motor roots of the cauda equina between these levels. Similarly motor latencies can be recorded from spinal stimulation to the anterior tibial muscles (L4 and L5 motor roots). Transrectal stimulation of the pudendal nerves is used to measure the pudendal nerve terminal motor latency. In 32 control subjects, matched for age and sex, mean motor latencies from L1 and L4 spinal stimulation were 5.5 +/- 0.4 ms and 4.4 +/- 0.4 ms (mean + SD). In the 10 patients with cauda equina disease including ependymoma, spinal stenosis, arachnoiditis and trauma, these latencies were 7.2 +/- 0.8 ms and 4.6 +/- 0.9 ms, a significant increase in the L1 latency. The L1/L4 latency ratios to the puborectalis muscle were 1.36 +/- 0.09 in control subjects and 1.72 +/- 0.13 in cauda equina patients. Pudendal nerve terminal motor latencies were normal in eight of the 10 patients with cauda equina disease. The single fibre EMG fibre density in the external and sphincter muscle (normal, 1.5 +/- 0.16) was increased in patients with cauda equina lesions (1.73 +/- 0.28), but was increased more than two standard deviations from the mean only in three patients. This increase in fibre density was not of diagnostic value since it was also found in two of the four patients with low back pain. Slowing of motor conduction in the cauda equina is thus a useful indication of damage to these intraspinal motor roots. These investigations can be used in the selection of patients for myelography, and to follow progress in patients managed conservatively.


Protection of BNT162b2 Vaccine Booster against Covid-19 in Israel.
On July 30, 2021, the administration of a third (booster) dose of the BNT162b2 messenger RNA vaccine (Pfizer-BioNTech) was approved in Israel for persons who were 60 years of age or older and who had received a second dose of vaccine at least 5 months earlier. Data are needed regarding the effect of the booster dose on the rate of confirmed coronavirus 2019 disease (Covid-19) and the rate of severe illness. We extracted data for the period from July 30 through August 31, 2021, from the Israeli Ministry of Health database regarding 1,137,804 persons who were 60 years of age or older and had been fully vaccinated (i.e., had received two doses of BNT162b2) at least 5 months earlier. In the primary analysis, we compared the rate of confirmed Covid-19 and the rate of severe illness between those who had received a booster injection at least 12 days earlier (booster group) and those who had not received a booster injection (nonbooster group). In a secondary analysis, we evaluated the rate of infection 4 to 6 days after the booster dose as compared with the rate at least 12 days after the booster. In all the analyses, we used Poisson regression after adjusting for possible confounding factors. At least 12 days after the booster dose, the rate of confirmed infection was lower in the booster group than in the nonbooster group by a factor of 11.3 (95% confidence interval [CI], 10.4 to 12.3); the rate of severe illness was lower by a factor of 19.5 (95% CI, 12.9 to 29.5). In a secondary analysis, the rate of confirmed infection at least 12 days after vaccination was lower than the rate after 4 to 6 days by a factor of 5.4 (95% CI, 4.8 to 6.1). In this study involving participants who were 60 years of age or older and had received two doses of the BNT162b2 vaccine at least 5 months earlier, we found that the rates of confirmed Covid-19 and severe illness were substantially lower among those who received a booster (third) dose of the BNT162b2 vaccine.


alpha-Melanocyte-stimulating hormone protects from ultraviolet radiation-induced apoptosis and DNA damage.
Ultraviolet radiation is a well established epidemiologic risk factor for malignant melanoma. This observation has been linked to the relative resistance of normal melanocytes to ultraviolet B (UVB) radiation-induced apoptosis, which consequently leads to accumulation of UVB radiation-induced DNA lesions in melanocytes. Therefore, identification of physiologic factors regulating UVB radiation-induced apoptosis and DNA damage of melanocytes is of utmost biological importance. We show that the neuropeptide alpha-melanocyte-stimulating hormone (alpha-MSH) blocks UVB radiation-induced apoptosis of normal human melanocytes in vitro. The anti-apoptotic activity of alpha-MSH is not mediated by filtering or by induction of melanin synthesis in melanocytes. alpha-MSH neither leads to changes in the cell cycle distribution nor induces alterations in the expression of the apoptosis-related proteins Bcl(2), Bcl(x), Bax, p53, CD95 (Fas/APO-1), and CD95L (FasL). In contrast, alpha-MSH markedly reduces the formation of UVB radiation-induced DNA damage as demonstrated by reduced amounts of cyclobutane pyrimidine dimers, ultimately leading to reduced apoptosis. The reduction of UV radiation-induced DNA damage by alpha-MSH appears to be related to induction of nucleotide excision repair, because UV radiation-mediated apoptosis was not blocked by alpha-MSH in nucleotide excision repair-deficient fibroblasts. These data, for the first time, demonstrate regulation of UVB radiation-induced apoptosis of human melanocytes by a neuropeptide that is physiologically expressed within the epidermis. Apart from its ability to induce photoprotective melanin synthesis, alpha-MSH appears to exert the capacity to reduce UV radiation-induced DNA damage and, thus, may act as a potent protection factor against the harmful effects of UV radiation on the genomic stability of epidermal cells.


Linear transformations of data space in MEG.
Magnetoencephalography (MEG) is a method which allows the non-invasive measurement of the minute magnetic field which is generated by ion currents in the brain. Due to the complex sensitivity profile of the sensors, the measured data are a non-trivial representation of the currents where information specific to local generators is distributed across many channels and each channel contains a mixture of contributions from many such generators. We propose a framework which generates a new representation of the data through a linear transformation which is designed so that some desired property is optimized in one or more new virtual channel(s). First figures of merit are suggested to describe the relation between the measured data and the underlying currents. Within this context the new framework is established by first showing how the transformation matrix itself is designed and then by its application to real and simulated data. The results demonstrate that the proposed linear transformations of data space provide a computationally efficient tool for analysis and a very much needed dimensional reduction of the data.


Sympatric Parasites Have Similar Host-Associated, but Asynchronous, Patterns of Diversification.
Parasitism is a common symbiotic interaction across diverse natural systems. Using a comparative evolutionary approach, we investigated the contributions of both host phylogeny and abiotic factors toward diversification of phylogenetically independent endoparasites that inhabit essentially the same physical space. We tested for host-parasite and parasite-parasite phylogenetic concordance in western North American chipmunks (Rodentia: Sciuridae) and two distantly related species of pinworms (Nematoda: Oxyurida). Deep structure in molecular phylogenies revealed signals of host-associated divergence in both parasite species, while shallower phylogeographic structure varied between the two parasites. This suggests that although these parasites experienced similar landscapes and cyclic climate processes, temporally distinctive diversification events were associated with differences in the initiation of their association with host lineages. When climate cycles initiate diversification, partially congruent, but asynchronous, host-associated parasite phylogenies may emerge.


Lung-resistance-related protein expression is a negative predictive factor for response to conventional low but not to intensified dose alkylating chemotherapy in multiple myeloma.
This study was undertaken to assess the significance of lung-resistance related protein (LRP) expression in plasma cells from untreated multiple myeloma (MM) patients and to determine whether LRP was associated with a poor response and survival in patients treated with different dose regimens of melphalan. Seventy untreated patients received conventional oral dose melphalan (0.25 mg/kg, day 1 to 4) combined with prednisone (MP) or intravenous intermediate-IDM; 70 mg/m2) or high- (140 mg/m2) dose Melphalan (HDM). LRP expression was assessed with immunocytochemistry using the LRP-56 monoclonal antibody. LRP expression was found in 47% of patients. In the MP treated patients, LRP expression was a significant prognostic factor regarding response induction (P < .05), event free survival (P < .003), and overall survival (P < .001). In the intensified dose melphalan treated patients LRP did not have a prognostic value. The response rates of LRP-positive patients to MP and IDM/HDM were 18% versus 81%, respectively (P < .0001). We conclude that LRP is frequently expressed in untreated MM patients and is an independent predictor for response and survival in patients treated with MP. Pretreatment assessment of LRP identifies a subpopulation of patients with a poor probability of response to conventional dose melphalan. Dose intensification of melphalan is likely to overcome LRP-mediated resistance.