Pattern Recognition via Observation Correlations.
In some pattern recognition tasks multiple observations of an observation vector Y = {Y1, Y2, ..., YM} are available for each object and the covariances of the Yi are characteristic of the object. With the assistance of a model of the generating process for Y a theoretical basis for the comparison of the covariance matrices is developed. Measurements based on synthetic data support the theory, and an application to some speaker verification is given as an example.


Results of aortic grafting in occlusive vs aneurysmal disease.
Recent demographic and biochemical studies have suggested that there may be significant differences in the population of patients undergoing reconstruction for an aortic aneurysm (AA) or atherosclerotic occlusive disease (OCC) of the abdominal aorta. The purpose of this retrospective study was to compare the results of aortic surgery in these different patient groups. One hundred three patients undergoing aortic grafting were reviewed. Twenty-four patients underwent reconstruction for OCC, 65 selectively for asymptomatic aneurysms (EAA), and 14 for ruptured aortic aneurysms (RAA). The mean age for OCC patients (54 years) was significantly lower than that for EAA (68 years) or RAA patients (71 years). Patients with AA were predominantly male (92%) compared with those with OCC (62%). Despite the age and sex differences, there were no significant differences between the groups in the incidence of cardiac disease, hypertension, smoking history, diabetes mellitus, or renal or pulmonary disease. In addition, there were no significant intraoperative differences in terms of operative and clamp times, fluid administration, urine output, hypotension, or subsequent postoperative morbidity and mortality. We conclude that despite AA and OCC patients being demographically distinct, there is essentially no difference in elective aortic bifurcation graft surgery between these two populations.


Prolonged duration of craving, mood, and autonomic responses elicited by cues and imagery in smokers: Effects of tobacco deprivation and sex.
Increases in self-reported craving and changes in autonomic functioning are reliably elicited when smokers are exposed to tobacco-related stimuli compared with neutral stimuli. However, few studies have reported the time course of cue-elicited craving or have directly compared the effectiveness of smoking cues versus imagery to evoke a craving response. In addition to these two issues, we investigated the influence of tobacco deprivation and sex on craving, mood, and autonomic responses. Sixty cigarette smokers (30 men, 30 women) were tested in two counterbalanced sessions, one after overnight tobacco deprivation and one during ad libitum smoking. At each session, participants were exposed to four randomized experimental trials: smoking imagery, neutral imagery, smoking cues, and neutral cues. Tobacco craving and mood were assessed repeatedly and physiological measures were recorded continuously for 30 min after imagery or cue exposure. Compared with neutral trials, smoking cues and smoking imagery reliably increased tobacco craving, negative mood, heart rate, and blood pressure and decreased positive mood ratings. Changes were observed immediately after cue and imagery presentation and remained unchanged for 30 min. Responding was greater in the nondeprived condition, and cues elicited more robust responding than imagery for most measures. Women responded more robustly to smoking cues only in the nondeprived condition, whereas imagery evoked greater responses in men during both conditions. These findings provide new data on the time course, magnitude, and tobacco deprivation effects on elicited craving. Sex differences were dependent on stimulus type and deprivation condition.


Assembly of von Willebrand factor tubules with in vivo helical parameters requires A1 domain insertion.
The von Willebrand factor (VWF) glycoprotein is stored in tubular form in Weibel-Palade bodies (WPBs) before secretion from endothelial cells into the bloodstream. The organization of VWF in the tubules promotes formation of covalently linked VWF polymers and enables orderly secretion without polymer tangling. Recent studies have described the high-resolution structure of helical tubular cores formed in vitro by the D1D2 and D'D3 amino-terminal protein segments of VWF. Here we show that formation of tubules with the helical geometry observed for VWF in intracellular WPBs requires also the VWA1 (A1) domain. We reconstituted VWF tubules from segments containing the A1 domain and discovered it to be inserted between helical turns of the tubule, altering helical parameters and explaining the increased robustness of tubule formation when A1 is present. The conclusion from this observation is that the A1 domain has a direct role in VWF assembly, along with its known activity in hemostasis after secretion.


Effect of fixation time on breast biomarker expression: a controlled study using cell line-derived xenografted (CDX) tumours.
Altering the length of time specimens are placed in fixative without compromising analytical testing accuracy is a continuous challenge in the anatomical pathology lab. The aim of this study was to determine under controlled conditions the effects of variable fixation time on breast biomarker expression in human breast cancer cell line-derived xenografted (CDX) tumours. CDX tumours using strong oestrogen receptor (ER)-positive, Her2-negative (MCF7) and weak ER-positive, Her2 equivocal (T47D) breast cancer cell lines were fixed for various times ranging from 1 to 336 hours in 10% neutral buffered formalin. CDX tumours were processed according to routine biomarker testing protocols and stained for ER and Her2 immunohistochemistry (IHC) and processed for HER2 fluorescence in situ hybridisation (FISH). The tumours were evaluated using Allred scoring for ER and current ASCO/CAP guidelines for Her2, and by objective cell counting methodology. No differences were found in expression of ER in either MCF7 or T47D CDX tumours under variable fixation. T47D tumours displayed equivocal Her2 staining when fixed for 24 hours, but fixation for ≤8 hours resulted in consistently negative staining while tumours fixed for >72 hours demonstrated consistent equivocal staining (p<0.01). Cell counting assays revealed only a significant increase in sensitivity in tumours fixed for >72 hours (p<0.01). As expected, FISH results were unaffected by variable fixation. Neither shortened nor prolonged fixation affects ER expression, consistent with previous findings. In equivocal Her2-expressing tumours, however, increasing fixation increased the sensitivity of Her2 IHC reporting while not affecting FISH.


Experiences and Lessons Learned in Developing and Implementing a Population-Based Nutrition and Health Surveillance System in Guatemala 2011-2021.
Practice-based experiences documenting development and implementation of nutrition and health surveillance systems are needed. To describe processes, methods, and lessons learned from developing and implementing a population-based household nutrition and health surveillance system in Guatemala. The phases and methods for the design and implementation of the surveillance system are described. Efforts to institutionalize the system in government institutions are described, and illustrative examples describing different data uses, and lessons learned are provided. After initial assessments of data needs and consultations with officials in government institutions and partners in the country, a population-based nutrition surveillance system prototype with complex sampling was designed and tested in 5 Guatemalan Highland departments in 2011. After dissemination of the prototype, government and partners expanded the content, and multitopic nutrition and health surveillance cycles were collected in 2013, 2015, 2016, 2017/18, and 2018/19 providing nationally representative data for households, women of reproductive age (15-49 y), and children aged 0-59 mo. For each cycle, data were to be collected from 100 clusters, 30 households in each, and 1 woman and 1 child per household. Content covered ∼25 health and nutrition topics, including coverage of all large-scale nutrition-specific interventions; the micronutrient content of fortifiable sugar, salt, and bread samples; anthropometry; and biomarkers to assess annually, or at least once, ∼25 indicators of micronutrient status and chronic disease. Data were collected by 3-5 highly trained field teams. The design was flexible and revised each cycle allowing potential changes to questionnaires, population groups, biomarkers, survey design, or other changes. Data were used to change national guidelines for vitamin A and B-12 interventions, among others, and evaluate interventions. Barriers included frequent changes of high-level government officials and heavy dependence on US funding. This system provides high-quality data, fills critical data gaps, and can serve as a useful model for others.


Contraceptive use among adolescent mothers at 6 months postpartum.
To assess patterns and predictors of reliable and unreliable contraceptive use among adolescent mothers in the first 6 months following delivery. We surveyed 462 women, 18 years of age or younger, at delivery and again at 6 months postpartum. Contraceptive behaviors were evaluated among the 359 adolescents who stated they were sexually active and not trying to conceive. Method discontinuation and switching were common during the 6-month interval. Only 100 of 189 adolescents (53%) initially prescribed oral contraceptives were still using this method 6 months after delivery; ten of these 100 stated that they had missed at least three pills in the last cycle. Twelve (10%) of the 115 adolescents who initiated depot-medroxyprogesterone acetate failed to obtain a second injection within 4 months of the initial injection or use an alternative method. In contrast nine of the ten women who received levonorgestrel implants were still using this method 6 months after delivery. Overall, 76% of the sample reported using reliable contraception at last intercourse. Multivariate analyses identified seven factors as predictive of reliable contraceptive use: school enrollment, not having failed a grade in school, adequate support, belief that pregnancy is likely without birth control, attendance at postpartum visit, prior abortion, and the adolescent's desire to wait at least 2 years before having another child. Interventions designed to reduce rapid repeat pregnancy during the adolescent years should address emotional, financial, and educational, as well as contraceptive, needs.


Microbial mat controls on infaunal abundance and diversity in modern marine microbialites.
Microbialites are the most abundant macrofossils of the Precambrian. Decline in microbialite abundance and diversity during the terminal Proterozoic and early Phanerozoic has historically been attributed to the concurrent radiation of complex metazoans. Similarly, the apparent resurgence of microbialites in the wake of Paleozoic and Mesozoic mass extinctions is frequently linked to drastic declines in metazoan diversity and abundance. However, it has become increasing clear that microbialites are relatively common in certain modern shallow, normal marine carbonate environments-foremost the Bahamas. For the first time, we present data, collected from the Exuma Cays, the Bahamas, systematically characterizing the relationship between framework-building cyanobacteria, microbialite fabrics, and microbialite-associated metazoan abundance and diversity. We document the coexistence of diverse microbialite and infaunal metazoan communities and demonstrate that the predominant control upon both microbialite fabric and metazoan community structure is microbial mat type. These findings necessitate that we rethink prevalent interpretations of microbialite-metazoan interactions and imply that microbialites are not passive recipients of metazoan-mediated alteration. Additionally, this work provides support for the theory that certain Precambrian microbialites may have been havens of early complex metazoan life, rather than bereft of metazoans, as has been traditionally envisaged.


Molecular detection and characterization of Leishmania infantum in free-ranging Egyptian mongoose (Herpestes ichneumon).
Wild mammals are susceptible to infection by Leishmania parasites. Although canine leishmaniasis is widely distributed in mainland Portugal, the sylvatic cycle of the parasite remains poorly understood. In this study, the occurrence of L. infantum in wild carnivores from Portugal was assessed by molecular screening of 132 hunted or accidentally road-killed animals. Spleen samples from Egyptian mongoose, red fox, stone marten, common genet and European badger were tested by amplification of Leishmania kinetoplastid DNA and ITS1. Five egyptian mongoose were confirmed Leishmania DNA-positive by kDNA-PCR. Phylogenetic analysis of a kDNA amplicon sequence clustered the strain with L. infantum sequences from Portugal. These results may suggest that L. infantum strains circulating in wild animals are genetically related with strains from more humanized settings. Exposure of wild carnivores to Leishmania infantum emphasizes the need of systematic studies to clarify the role of several taxa in the eco-epidemiology of leishmaniasis in Portugal, particularly in areas of carnivore species synanthropy and wherein disease control in the domestic population is inefficient or insufficient.


Lottery spending: a non-parametric analysis.
We analyze the spending of individuals in the United States on lottery tickets in an average month, as reported in surveys. We view these surveys as sampling from an unknown distribution, and we use non-parametric methods to compare properties of this distribution for various demographic groups, as well as claims that some properties of this distribution are constant across surveys. We find that the observed higher spending by Hispanic lottery players can be attributed to differences in education levels, and we dispute previous claims that the top 10% of lottery players consistently account for 50% of lottery sales.


Immunogenic properties of archaeal species found in bioaerosols.
The etiology of bioaerosol-related pulmonary diseases remains poorly understood. Recently, archaea emerged as prominent airborne components of agricultural environments, but the consequences of airway exposure to archaea remain unknown. Since subcomponents of archaea can be immunogenic, we used a murine model to study the pulmonary immune responses to two archaeal species found in agricultural facilities: Methanobrevibacter smithii (MBS) and Methanosphaera stadtmanae (MSS). Mice were administered intranasally with 6.25, 25 or 100 µg of MBS or MSS, once daily, 3 days a week, for 3 weeks. MSS induced more severe histopathological alterations than MBS with perivascular accumulation of granulocytes, pronounced thickening of the alveolar septa, alveolar macrophages accumulation and increased perivascular mononucleated cell accumulation. Analyses of bronchoalveolar lavage fluids revealed up to 3 times greater leukocyte accumulation with MSS compared to MBS. Instillation of 100 µg of MBS or MSS caused predominant accumulation of monocyte/macrophages (4.5×10(5) and 4.8×10(5) cells/ml respectively) followed by CD4(+) T cells (1.38×10(5) and 1.94×10(5) cells/ml respectively), B cells (0.73×10(5) and 1.28×10(5) cells/ml respectively), and CD8(+) T cells (0.20×10(5) and 0.31×10(5) cells/ml respectively) in the airways. Both archaeal species induced similar titers of antigen-specific IgGs in plasma. MSS but not MBS caused an accumulation of eosinophils and neutrophils in the lungs, which surprisingly, correlated inversely with the size of the inoculum. Stronger immunogenicity of MSS was confirmed by a 3 fold higher accumulation of myeloid dendritic cells in the airways, compared to MBS. Thus, the dose and species of archaea determine the magnitude and nature of the pulmonary immune response. This is the first report of an immunomodulatory role of archaeal species found in bioaerosols.


Development of a method for detection of human rotavirus in water and sewage.
The simian rotavirus SA11 was used to develop a simple, reliable, and efficient method to concentrate rotavirus from tap water, treated sewage, and raw sewage by absorption to and elution from Filterite fiberglass-epoxy filters. SA11 adsorbed optimally to Filterite filters from water containing 0.5 mM AlCl3 at pH 3.5. Filter-bound virus was eluted with 0.05 M glycine-NaOH supplemented with 10% tryptose phosphate broth at pH 10. SA11 was quantitated by plaque assay, whereas human rotavirus was detected by immunofluorescence. The method was applied to detect rotavirus in raw and treated sewage at two Houston, Tex., sewage treatment plants. The sewage isolates were identified as rotavirus, probably a human strain, based on several criteria. The sewage isolates were detectable by an immunofluorescence test, using anti-SA11 serum which would detect the simian, human bovine, and porcine rotaviruses. No reaction was noted by immunofluorescence with the reoviruses or several common enteroviruses. The sewage isolates were neutralized by convalescent sera from a human adult and infant who had been infected by rotavirus as well as by a hyperimmune serum prepared in guinea pigs against purified human rotavirus. Preimmune or preillness sera did not react with the isolates by neutralization or immunofluorescence. The natural isolates were sensitive to pH 11 and other inactivating agents, similar to SA11. The buoyant density of the sewage isolates in CsCl gradients was 1.36 g/cm3, which is the value usually reported for complete, infectious rotavirus particles. The double-shelled particle diameter was 67.1 +/- 2.4 nm. Finally, electron micrographs of cell lysates inoculated with the sewage isolate showed particles displaying characteristic rotavirus morphology.


Brain Metastases and Leptomeningeal Disease.
Central nervous system (CNS) metastases include brain parenchymal, spinal cord, and leptomeningeal metastases. This article discusses the diagnostic and therapeutic advances of the last decade that have improved outcomes for patients with these CNS metastases. The diagnostic tools for CNS metastases, particularly leptomeningeal disease, have evolved over the past decade with respect to advancements in CSF analysis. Multiple medical therapies are now available for brain metastasis treatment that have shown CNS efficacy, including targeted therapies and antibody-drug conjugates. Molecular testing for CNS metastases has become more common and the repertoire of molecularly targeted therapies continues to expand. Advancements in radiation therapy, including improvements in stereotactic radiation techniques, whole-brain radiation with hippocampal avoidance, and proton beam radiation, have changed the radiation management of patients with CNS metastases. New intrathecal agents are currently being tested for the management of leptomeningeal metastases. CNS metastases are far more common than primary brain tumors and are increasing in prevalence in the setting of improved treatments and prolonged survival in patients with systemic cancers. There have been many changes in the diagnostics and treatment of CNS metastases, yielding subsequent improvements in patient outcomes with further advancements on the horizon.


[Tactics of treating children with Coat's disease].
Fifty children with Coats disease, aged 2 months to 12 years (mean age 8.25 +/- 2.72 years), were observed. Four stages are distinguished in the disease course: initial, moderate, advanced, and terminal. Treatment including extensive retinal argon laser coagulation, cryotherapy, scleral bucking, and subretinal liquid draining was carried out in 32 children (33 eyes). Stable anatomic results were attained in 97% eyes. Visual acuity of at least 0.02 was retained in 87.9% children.


Defining species specific genome differences in malaria parasites.
In recent years a number of genome sequences for different plasmodium species have become available. This has allowed the identification of numerous conserved genes across the different species and has significantly enhanced our understanding of parasite biology. In contrast little is known about species specific differences between the different genomes partly due to the lower sequence coverage and therefore relatively poor annotation of some of the draft genomes particularly the rodent malarias parasite species. To improve the current annotation and gene identification status of the draft genomes of P. berghei, P. chabaudi and P. yoelii, we performed genome-wide comparisons between these three species. Through analyses via comparative genome hybridizations using a newly designed pan-rodent array as well as in depth bioinformatics analysis, we were able to improve on the coverage of the draft rodent parasite genomes by detecting orthologous genes between these related rodent parasite species. More than 1,000 orthologs for P. yoelii were now newly associated with a P. falciparum gene. In addition to extending the current core gene set for all plasmodium species this analysis also for the first time identifies a relatively small number of genes that are unique to the primate malaria parasites while a larger gene set is uniquely conserved amongst the rodent malaria parasites. These findings allow a more thorough investigation of the genes that are important for host specificity in malaria.


Gene-expression profiling of White spot syndrome virus in vivo.
White spot syndrome virus, type species of the genus Whispovirus in the family Nimaviridae, is a large, double-stranded DNA (dsDNA) virus that infects crustaceans. The genome of the completely sequenced isolate WSSV-TH encodes 184 putative open reading frames (ORFs), the functions of which are largely unknown. To study the transcription of these ORFs, a DNA microarray was constructed, containing probes corresponding to nearly all putative WSSV-TH ORFs. Transcripts of 79 % of these ORFs could be detected in the gills of WSSV-infected shrimp (Penaeus monodon). Clustering of the transcription profiles of the individual genes during infection showed two major classes of genes: the first class reached maximal expression at 20 h post-infection (p.i.) (putative early) and the other class at 2 days p.i. (putative late). Nearly all major and minor structural virion-protein genes clustered in the latter group. These data provide evidence that, similar to other large, dsDNA viruses, the WSSV genes at large are expressed in a coordinated and cascaded fashion. Furthermore, the transcriptomes of the WSSV isolates WSSV-TH and TH-96-II, which have differential virulence, were compared at 2 days p.i. The TH-96-II genome encodes 10 ORFs that are not present in WSSV-TH, of which at least seven were expressed in P. monodon as well as in crayfish (Astacus leptodactylus), suggesting a functional but not essential role for these genes during infection. Expression levels of most other ORFs shared by both isolates were similar. Evaluation of transcription profiles by using a genome-wide approach provides a better understanding of WSSV transcription regulation and a new tool to study WSSV gene function.


Cancellous grafting and external fixation for unstable Colles' fractures.
We present a prospective study of the treatment of 32 unstable Colles' fractures by external fixation and cancellous grafting with minimal exposure. We inserted an external fixator between the radius and the second metacarpal, and maintained ligamentotaxis for five weeks. In 27 patients the result was good or excellent, but five fractures healed with malunion. All patients made a satisfactory functional recovery. At a mean follow-up of three years (1 to 5) after injury none had pain in the wrist and all were satisfied with the result. The average grip strength was 95% of normal. Seven patients had algodystrophy with mild impairment of finger movements in four. We conclude that the combination of cancellous grafting and external fixation is effective for the treatment of unstable Colles' fractures.


Quantifying Western blots: pitfalls of densitometry.
Although Western blots are frequently quantified, densitometry is not documented and appears to be based merely on traditions and guesswork. Confirming previous experience, none of 100 randomly selected and systematically scanned most recent papers provided sufficient information on how Western blot results were translated into statistical values. The importance of such information, however, becomes evident from our correlations of plasma erythropoietin values of various mammals determined using RIA and Western blot densitometry. Different common densitometry procedures applied to the identical Western blot revealed p-values of these correlations ranging from 0.000013 to 0.76 reflecting the necessity of a scientifically sound basis for densitometry of Western blots. At present, the current lack of any definitions in densitometry opens the door to uncontrollable acquisition of any desired p-value. Here we provide data that define what should be considered, what avoided and what documented when quantifying Western blots.


Seasonal migration and health care.
This article examines health care use and issues among seasonal migrants in an urban setting in the Sunbelt. The study, based on a survey of 230 American and Canadian snowbirds in Phoenix, Arizona, complements previous research on health care use among Canadian snowbirds in Florida. Our results show that health care use among seasonal migrants in Phoenix is substantial, and it varies by citizenship (American vs. Canadian), age, number of winter visits, and length of winter stay. We argue that health care use is symptomatic of social and emotional attachments to the winter residence that develop and deepen over a period of years. Phoenix snowbirds view their lifestyles as healthy, they are adept in adjusting to health decrements, and the majority plan to continue spending time in both their summer and winter residences as long as their health permits.


Effects of Chest Compression on Ventilation Quality during Cardiopulmonary Resuscitation.
Ventilation is an important part of cardiopulmonary resuscitation (CPR). The advanced airway mode and 30:2 mode are used for intubated and non-intubated patients, respectively. It is debatable that passive produced by 30 compressions can provide adequate tidal volume for 30:2 mode. In addition, the fragmented ventilation caused by continuous compression may result in ineffective ventilation. In the study, one pig was anaesthetized and intubated for 2 CPRs. Continuous chest compressions with ventilation and continuous chest compressions without mechanical ventilation were performed in 2 CPRs, respectively. Three 10-minute data segments including a period of normal ventilation (V segment), a period of only compressions without ventilation (C segment), and a period of compressions with ventilation (C-V segment) were used to analyze peek flow (PF), peek pressure (PP) and tidal volume. All the data was presented as mean ± standard deviation. Chest compression resulted in 14.90% increase in mean PP (2401.40 ± 94.75 Pa vs 2822.06 ± 291.10 Pa, p<0.05), 81.46% increase in average PF (319.58 ± 56.93 ml/s vs 579.92 ± 80.27 ml/s, p<0.05). The mean tidal volumes for C segment, V segment and C-V segment were 189.13 ml, 514.72 ml, and 429.26ml, respectively. Continuous compressions reduced the accumulative tidal volume, but when five compressions were made in one inspiratory phase, there is almost no loss of tidal volume (510.86 ± 47.24 ml vs 514.72 ± 29.25 ml, p<0.05). The study suggested the ventilator without feedback regulation might reduce the peek pressure during CPR and 5 compressions in 2 s inspiratory phase provided higher tidal volume.Clinical Relevance- This study shows that 150 chest compressions per minute provided greater tidal volume than 100 and 120 compressions per minute; continuous chest compressions could also provide a certain amount of oxygen supply.


Longitudinal assessment of vibrations during manual and power wheelchair driving over select sidewalk surfaces.
Wheelchair users rely on their wheelchairs for mobility for extended periods of time every day. According to the International Standards Organization 2631-1 standard on human vibration, individuals in a seated position when exposed to whole-body vibrations (WBV) are at risk of injury. This study evaluated vibration exposure during manual and power wheelchair driving over nine sidewalk surfaces and differences in vibration exposure over 3 years. Ten nondisabled subjects were asked to drive a manual wheelchair at 1 m/s and a power wheelchair at 1 m/s and 2 m/s over nine sidewalk surfaces while WBV were measured at the seat and footrest of the wheelchair. At 1 m/s, significant differences existed between surfaces and years at both the seat and the footrest for the manual and power wheelchair users. At 2 m/s, significant differences existed between surfaces and years at the seat and the footrest for power wheelchair users. Our results show that both manual and power wheelchair users may be at risk for secondary injuries from WBV when traveling over certain surfaces.


[Antioxidant therapy with xymedon of patients with chronic osteomyelitis].
An investigation of the antioxidant activity of blood plasma allowed a conclusion to be made of an alteration of the free radical processes in patients with chronic osteomyelitis. Experimental data on the influence of the antioxidants Xymedon and Methyluracil on the main free radicals, lipid peroxidation and chemiluminescence of peritoneal macrophages of rats allowed the substantiation of using the medicines in the complex treatment of patients in the pre- and postoperative periods. As a result, in 25 patients Xymedon improved the blood indices, increased antioxidant activity of plasma and reduced the amount of recurrences of the disease as compared with the patients taking Methyluracil.


The Relationship between MRI Radiofrequency Energy and Function of Nonconditional Implanted Cardiac Devices: A Prospective Evaluation.
Background The risks associated with MRI in individuals who have implanted cardiac devices are thought to arise from the interaction between the implanted device and static, gradient, and radiofrequency magnetic fields. Purpose To determine the relationship between the peak whole-body averaged specific absorption rate (SAR) and change in magnetic field per unit time (dB/dt), maximum specific energy dose, imaging region, and implanted cardiac device characteristics and their function in patients undergoing MRI. Materials and Methods This prospective observational cohort study was conducted from October 16, 2003, to January 22, 2015 (https://ClinicalTrials.gov, NCT01130896). Any individual with an implanted cardiac device who was referred for MRI was included. Clinical MRI protocols without SAR restriction were used. Exclusion criteria were newly implanted leads, abandoned or epicardial leads, and dependence on a pacemaker with an implantable cardioverter defibrillator without asynchronous pacing capability. For each MRI pulse sequence, the calculated whole-body values for SAR, dB/dt, and scan duration were collected. Atrial and ventricular sensing, lead impedance, and capture threshold were evaluated before and immediately after (within 10 minutes) completion of each MRI examination. Generalized estimating equations with Gaussian family, identity link, and an exchangeable working correlation matrix were used for statistical analysis. Results A total of 2028 MRI examinations were performed in 1464 study participants with 2755 device leads (mean age, 67 years ± 15 [standard deviation]; 930 men [64%]). There was no evidence of an association between radiofrequency energy deposition, dB/dt, or scan duration and changes in device parameters. Thoracic MRI was associated with decreased battery voltage immediately after MRI (β = -0.008 V, P < .001). Additionally, right ventricular (RV) lead length was associated with decreased RV sensing (β = -0.012 mV, P = .05) and reduced RV capture threshold (β = -0.002 V, P < .01) immediately after MRI. Conclusion There was no evidence of an association between MRI parameters that characterize patient exposure to radiofrequency energy and changes in device and lead parameters immediately after MRI. Nevertheless, device interrogation before and after MRI remains mandatory due to the potential for device reset and changes in lead or generator parameters. © RSNA, 2020 See also the editorial by Shellock in this issue.


Structural-functional organization of the main light harvesting complex and photosystem 2 of higher plants.
The interrelation between spectral and structural-functional properties of LhcIIb was studied. The dipole strength of the main Q(y )bands of chlorophylls (Chl a 30.8 D(2); Chl b 18.5 D(2)) and chlorophyll a/b ratio (Chl a/Chl b = 7 : 6) were determined for LhcIIb. The Chl a/Chl b value shows that the subunit of this complex contains seven Chl a and six Chl b molecules. Individual bands of chlorophylls (bands in stokes and anti-Stokes parts at 77 K were Lorentzian and Gaussian, respectively) were resolved using synchronized deconvolution of absorption, CD, and LD bands of chlorophylls. Seven of these bands belonged to Chl a. Parameters of absorption bands of Chl a indicate that seven molecules represent a united cluster (heptamer) with exciton interactions, determining the spectrum of LhcIIb in the Chl a absorption region. Parameters of absorption bands of Chl b show the existence of three clusters: monomer (639.6 nm), dimer (645.2 and 647.4 nm), and trimer (649.8 and 654.1 nm). These clusters and their properties agree with the well-known structure of porphyrin groups of the LhcIIb subunit (Kuhlbrandt, 1994). A distorted ring of seven porphyrins in the stromal range of the subunit corresponds to Chl a heptamer; a separately located molecule near the N-terminal domain on the stromal side of the subunit corresponds to Chl b monomer; a dimer and a trimer of porphyrins in the lumenal range of the subunit correspond to the dimer and trimer of Chl b, respectively. The calculated lifetimes of the excitation energy (exciton) transfer in subunit and trimer of LhcIIb confirm this location of pigments. The geometry of the Chl a heptamer (mutual orientation of transition dipole moments) was determined by the steady-state Kasha-Tinoco approximation using parameters of individual bands of exciton splitting. The calculated parameters of mutual orientation of Chl a dipoles agree with the topography of the stromal porphyrins found by electron crystallography (Kuhlbrandt, 1994). A structural model of the granal multicentral macrocomplex of PSII (MPSII) is suggested. The lifetimes of the exciton migration between the main pigment-protein compartments of MPSII were calculated. The results of calculation are consistent with the structural model of the photosystem. The location of pigments provides for fast exciton hopping between Chl a clusters of neighboring proteins in the MPSII along the stromal surface within the membrane (5-25 psec) and between stacked membranes (approximately 40 psec) of chloroplast grana.


Introduction of a waterless alcohol-based hand rub in a long-term-care facility.
To examine the impact of introduction of an alcohol-based hand rub on hand hygiene knowledge and compliance and hand colonization of healthcare workers (HCWs) in a long-term-care facility (LTCF). Two floors of an LTCF participated. Ward A used the hand rub as an adjunct to soap and water; ward B was the control. HCWs' hands were cultured using the bag-broth technique for Staphylococcus aureus, gram-negative bacilli (GNB), Candida, and vancomycin-resistant enterococci (VRE). HCWs completed a questionnaire at baseline and after an educational intervention and introduction of rub. Hand hygiene practices, knowledge, and opinions did not change after the educational or rub intervention. Ward A HCWs thought that the rub was faster (P = .002) and less drying (P = .04) than soap. Hand hygiene frequency did not differ at baseline between the two floors, but increased on ward A by the end of the study (P = .04). HCWs were colonized frequently with GNB (66%), Candida (41%), S. aureus (20%), and VRE (9%). Although colonization did not change from baseline on either ward, the rub was more effective in clearing GNB P =.03) and S. aureus (P = .003). Nosocomial infection rates did not change. The alcohol-based hand rub was a faster, more convenient, less drying method of hand hygiene for HCWs in an LTCF, and it improved compliance. Although microbial colonization did not change, the rub was more efficacious in removing pathogens already present on the hands of HCWs.


Color-image reconstruction for two-wavelength digital holography using a generalized phase-shifting approach.
We propose a color-image reconstruction method for two-wavelength digital holography using generalized phase-shifting digital holography (GPSDH). In this method, color interference fringes are captured by a digital camera with a Bayer array color filter, and phase shifting is simultaneously performed for all wavelengths. Color interference fringes are separated into three monochromatic interference fringes using a color-separation method that suppresses the color-filter crosstalk. The object wave is extracted from each monochromatic interference fringe using GPSDH, which prevents problems due to the phase shift wavelength dependence. Image reconstruction is performed using a shifted Fresnel transform-based method, in which the color reconstructed image is obtained by directly superposing the reconstructed images for all wavelengths. We verify the proposed method through optical experiments with a two-wavelength digital holography system. The results show that the dual-color image can be successfully reconstructed without chromatic aberration.


Pyrazolobenzothiazine-based carbothioamides as new structural leads for the inhibition of monoamine oxidases: design, synthesis, in vitro bioevaluation and molecular docking studies.
Two new series of pyrazolobenzothiazine-based carbothioamides (3a-o and 4a-o) were synthesized using saccharin as the starting material. The synthesized derivatives were investigated for their ability to inhibit monoamine oxidases (MAO). Compound 3b was found to be a very potent MAO-A inhibitor with an IC50 value of 0.003 ± 0.0007 μM, while compound 4d was the most effective inhibitor of MAO-B having an IC50 value of 0.02 ± 0.001 μM. Molecular docking studies were performed to identify the probable binding modes in the active site of the monoamine oxidase enzymes. The synthetic and computational investigations in the current work suggested that these newly identified inhibitors may serve as a powerful starting point for the exploration and optimization of potential therapeutic agents targeting Parkinson's disease.


Local adaptation when competition depends on phenotypic similarity.
Recent work incorporating demographic-genetic interactions indicates the importance of population size, gene flow, and selection in influencing local adaptation. This work typically assumes that density-dependent survival affects individuals equally, but individuals in natural population rarely compete equally. Among-individual differences in resource use generate stronger competition between more similar phenotypes (frequency-dependent competition) but it remains unclear how this additional form of selection changes the interactions between population size, gene flow, and local stabilizing selection. Here, we integrate migration-selection dynamics with frequency-dependent competition. We developed a coupled demographic-quantitative genetic model consisting of two patches connected by dispersal and subject to local stabilizing selection and competition. Our model shows that frequency-dependent competition slightly increases local adaptation, greatly increases genetic variance within patches, and reduces the amount that migration depresses population size, despite the increased genetic variance load. The effects of frequency-dependence depend on the strength of divergent selection, trait heritability, and when mortality occurs in the life cycle in relation to migration and reproduction. Essentially, frequency-dependent competition reduces the density-dependent interactions between migrants and residents, the extent to which depends on how different and common immigrants are compared to residents. Our results add new dynamics that illustrate how competition can alter the effects of gene flow and divergent selection on local adaptation and population carrying capacities.


The chemopreventive agent alpha-difluoromethylornithine blocks Ki-ras-dependent tumor formation and specific gene expression in Caco-2 cells.
Mutation of the Kirsten-ras (Ki-ras) proto-oncogene occurs frequently in colorectal cancers. alpha-Difluoromethylornithine (DFMO), an irreversible inhibitor of the polyamine biosynthetic enzyme, ornithine decarboxylase (ODC), inhibits Ki-ras transformation and colon tumorigenesis in carcinogen-treated animal models by mechanisms yet to be elucidated. Caco-2 cells transfected with an activated Ki-ras, but not parental cells, formed tumors in severe combined immunodeficient (SCID) mice. DFMO treatment (2% in drinking water) prevented tumor growth. Gene expression profiling was performed to identify Ki-ras-and DFMO-dependent patterns of gene expression. Microarray results were validated with real-time or semi-quantitative RT-PCR and/or Western blot analysis. Genes upregulated in Caco-2 cells expressing an activated Ki-ras encoded cytoskeletal-, transport-, protease-, and gap junction-associated proteins. These genes are important for normal development and maintenance of colonic epithelial tissue. Caco-2 cells transfected with an activated Ki-ras displayed increased expression of the integrin alpha 1 (INGA1) and enhanced cell migration on laminin. These parameters were unaffected by DFMO, but Ki-ras-dependent migration was inhibited by INGA1 antibodies. Other Ki-ras-dependent, but DFMO-independent, genes included transglutaminase (TGase) and kallikrein 6 (KLK6). Ki-ras-transfected cells also expressed increased levels of connexin43 (Cx43) (RNA and protein), tight junction protein, and endothelin 1. DFMO reversed these increases. The results indicated that the Ki-ras oncogene caused changes in experimental cell migration and cell-cell communication genes and that some of these changes could be reversed by DFMO.


Digital restoration of scintigraphic images by a two-step procedure.
A two-step procedure for restoring scintigraphic images is described. The first step uses a local-statistics algorithm for improvement in signal-to-noise ratio by smoothing the Poisson noise. The second step aims at improvement in spatial resolution by removing the linear blur using the iterative restoration algorithm. We present some results of computer simulations and restoration of scintigraphic images that demonstrate the effectiveness of the proposed procedure.


Molecular evolution of Pediculus humanus and the origin of clothing.
The human head louse (Pediculus humanus capitis) and body louse (P. humanus corporis or P. h. humanus) are strict, obligate human ectoparasites that differ mainly in their habitat on the host : the head louse lives and feeds exclusively on the scalp, whereas the body louse feeds on the body but lives in clothing. This ecological differentiation probably arose when humans adopted frequent use of clothing, an important event in human evolution for which there is no direct archaeological evidence. We therefore used a molecular clock approach to date the origin of body lice, assuming that this should correspond with the frequent use of clothing. Sequences were obtained from two mtDNA and two nuclear DNA segments from a global sample of 40 head and body lice, and from a chimpanzee louse to use as an outgroup. The results indicate greater diversity in African than non-African lice, suggesting an African origin of human lice. A molecular clock analysis indicates that body lice originated not more than about 72,000 +/- 42,000 years ago; the mtDNA sequences also indicate a demographic expansion of body lice that correlates with the spread of modern humans out of Africa. These results suggest that clothing was a surprisingly recent innovation in human evolution.


Strategic command, control, communications, and intelligence.
Command, control, communications, and intelligence (C(3)l) for nuclear forces are essential elements in the deterrence of nuclear war. The present C(3)l) system has vulnerabilities associated with its reliability, survivability, and endurance under attack, thereby weakening deterrence by increasing the ambiguity in our capabilities. Development of a reliable and enduring C(3)l) system would reduce this ambiguity. Its reliable, positive control of nuclear forces would give the national leadership more time to assess situations, ensure discriminate retaliation, and improve our ability to manage crises in general. These capabilities could help to stop a war rapidly should one start. A reliable and enduring C(3)1) system will be needed for a long time to come, even if a freeze on strategic nuclear forces is accomplished or other arms control successes achieved. Indeed, C(3)l) may be the best source today of confidence-building measures to reduce tensions and the threat of nuclear catastrophe.


Iron deposition in the corneal graft. Another corneal iron line.
Linear iron deposits in the corneal epithelium are well-documented phenomena in a variety of circumstances, both normal and pathologic. A characteristic annular pigmented line, appearing clinically as an "iron" line, occurred in the donor epithelium of approximately 13% of corneal grafts with retained sutures. Eight cases of normally functioning grafts had iron lines located just central to the sutures. These lines appeared at various intervals postoperatively and did not correlate with age or race. They were frequently found in association with other surface-related epithelial and subepithelial phenomena and were believed to be related to peripheral corneal topography and the lid-cornea interface. Although they are clinically iron, their precise histochemistry, cause, source, and natural history are yet to be determined.


Influence of nymphal Anax imperator (Odonata: Aeshnidae) on oviposition by the mosquito Culiseta longiareolata (Diptera: Culicidae) and community structure in temporary pools.
We assessed the overall (consumptive plus non-consumptive) and non-consumptive effects of nymphal Anax imperator (Aeshnidae: Odonata) on experimental pool communities by comparing three treatments: (1) control (no Anax); (2) free Anax (Anax was not constrained); and (3) caged Anax (Anax, enclosed within a cage, could not consume prey outside the cages, but could possibly influence them via perceived risk of predation). Fewer egg rafts of the mosquito Culiseta longiareolata were found in the free Anax treatment compared to the other two treatments. There was no statistically significant difference in the number of egg rafts between control and caged Anax pools. Thus, while Culiseta females apparently oviposit fewer egg rafts in the presence of unconstrained Anax, they did not respond to predation risk from the caged Anax. Larval Culiseta densities were drastically reduced by free Anax; there was nearly a 100% reduction in the number reaching metamorphosis (pupae) and a 100% reduction in emergence (pupal exuviae). There were no significant treatment effects on densities of crustaceans, Daphnia magna and Heterocypris sp., or on chironomid pupal exuviae. Ceratopogonid pupal abundance was higher in free Anax pools than in the other two treatments toward the end of the experiment. Free Anax caused a trophic cascade, presumably by strongly reducing the dominant periphyton grazer, Culiseta larvae; periphyton mass was greater in the free Anax treatment than in the control. However, there was no behavioral trophic cascade, i.e., no difference in periphyton abundance between the control and caged Anax treatment.


Forest loss and the biodiversity threshold: an evaluation considering species habitat requirements and the use of matrix habitats.
Habitat loss is the main driver of the current biodiversity crisis, a landscape-scale process that affects the survival of spatially-structured populations. Although it is well-established that species responses to habitat loss can be abrupt, the existence of a biodiversity threshold is still the cause of much controversy in the literature and would require that most species respond similarly to the loss of native vegetation. Here we test the existence of a biodiversity threshold, i.e. an abrupt decline in species richness, with habitat loss. We draw on a spatially-replicated dataset on Atlantic forest small mammals, consisting of 16 sampling sites divided between forests and matrix habitats in each of five 3600-ha landscapes (varying from 5% to 45% forest cover), and on an a priori classification of species into habitat requirement categories (forest specialists, habitat generalists and open-area specialists). Forest specialists declined abruptly below 30% of forest cover, and spillover to the matrix occurred only in more forested landscapes. Generalists responded positively to landscape heterogeneity, peaking at intermediary levels of forest cover. Open area specialists dominated the matrix and did not spillover to forests. As a result of these distinct responses, we observed a biodiversity threshold for the small mammal community below 30% forest cover, and a peak in species richness just above this threshold. Our results highlight that cross habitat spillover may be asymmetrical and contingent on landscape context, occurring mainly from forests to the matrix and only in more forested landscapes. Moreover, they indicate the potential for biodiversity thresholds in human-modified landscapes, and the importance of landscape heterogeneity to biodiversity. Since forest loss affected not only the conservation value of forest patches, but also the potential for biodiversity-mediated services in anthropogenic habitats, our work indicates the importance of proactive measures to avoid human-modified landscapes to cross this threshold.


Panoramic radiography and cone-beam computed tomography findings in preoperative examination of impacted mandibular third molars.
Preoperative radiographic examination of impacted mandibular third molars (IMTM) is essential to prevent inferior alveolar nerve injury during extraction. The purpose of this study was to evaluate the correlation between cone-beam computed tomography (CBCT) and digital panoramic radiography (DPR) findings in preoperative examination of IMTM. This retrospective study included 298 teeth in 191 individuals. The relationship between the inferior alveolar canal (IAC) and the IMTM (buccal, lingual, interradicular or inferior), the position of the IMTM with respect to the IAC (contact, no contact), the morphologic shape of the mandible in the IMTM region (round, lingual extended, lingual concave), the type of IMTM (vertical, horizontal or angular) and the number of roots of the IMTM were evaluated on CBCT images. DPR images were evaluated for the number of roots of the IMTM and for the most common radiographic findings indicating a relationship between the IAC and the IMTM (darkening of the roots, diversion of the IAC, narrowing of the IAC and interruption of the white line). Data were statistically analyzed with Cramer V coefficient, Kappa statistic, chi-square and Fisher's exact test. There was a significant difference in number of roots detected on DPR versus CBCT images. There was a significant association between the type of IMTM and the morphologic shape of the mandible on CBCT images. Darkening of the roots and interruption of the white line on DPR images were significantly associated with the presence of contact between the IMTM and the IAC on CBCT images. Panoramic radiography is inadequate, whereas CBCT is useful to detect multiple roots of IMTM. When darkening of the roots and interruption of the white line are observed on panoramic images, there is increased likelihood of contact between the IMTM and the IAC. CBCT is required in these cases.


Comparative Analysis of miRNA and EMT Markers in Metastatic Colorectal Cancer.
Colorectal cancer (CRC) is the fourth most commonly diagnosed malignant condition in the world. Micro RNAs (miRNAs) as well as epithelial to mesenchymal transition (EMT) play an important role in the pathogenesis of CRC. We performed a comparative analysis of the expression of selected miRNA genes and EMT markers in bioptic samples from patients (n = 45) with primary CRC or metastatic (m)CRC to the regional lymph node using reverse transcription-quantitative PCR and IHC staining. Results: Out of all miRNA analyzed, the miR-17 expression was most significantly different and associated with lower risk of CRC spread to the lymph node. In addition, significant relationships were found between the tumor side localization and several miRNAs expressions (miR-9, miR-29b, miR-19a, miR-19b, miR-21, miR-106a, miR-20a and miR-17). In addition, of the examined EMT markers, only VEGFA expression correlated with tumor progression (tumor grade G2). In the examined set of patient samples and their matched healthy tissue, several specific molecular markers (miRNAs associated with EMT and tumor progression) were identified with a promising prognostic potential. Their further examination in larger patient cohorts is planned to validate the present data.


Is vascular endothelial growth factor a useful biomarker in giant cell arteritis?
To assess the performance of circulating vascular endothelial growth factor (VEGF) levels as a tool for diagnosing giant cell arteritis (GCA) in a cohort of patients referred for assessment of suspected GCA. We selected 298 patients recruited to the multicentre study Temporal Artery Biopsy versus Ultrasound in diagnosis of suspected GCA (TABUL). In a random subset of 26 biopsy-proven GCA cases and 26 controls, serum from weeks 0, 2 and 26 was analysed for VEGF concentration using ELISA. VEGF concentration at week 0 was used to generate a receiver-operating characteristic curve and thereby identify a cut-off for an abnormal result which was used to analyse the full patient cohort. Sections of paraffin-embedded temporal artery were stained by immunohistochemistry for VEGF. The mean (95% CI) VEGF concentration at week 0 was 873 pg/mL (631 to 1110) in 26 patients versus 476 pg/mL (328 to 625) in 26 controls (p=0.017). This difference was not observed at any other time point. The optimal cut-off of 713 pg/mL was applied to the whole patient cohort (n=298), yielding sensitivity of 32% and specificity of 85%. This was not improved by combination with any clinical parameters. When patients with biopsy-proven GCA were compared with controls, sensitivity was 58% and specificity remained 85%. Sections of biopsy from biopsy-positive GCA showed intense staining in the adventitia which was not seen in controls. Serum VEGF concentration predicts biopsy positivity but is not useful for differentiating clinical cases of GCA from controls. Further studies into VEGF as a prognostic marker and therapeutic target are warranted. NCT00974883; Post-results.


Timing of Post-bath Skin Moisturizer Application to Newborn Infants: A Randomized Controlled Study.
To investigate whether the timing of postbath moisturizer application affected the skin moisture (SM) and body temperature (BT) of newborn infants. The researchers conducted a randomized controlled study with 80 newborns who were monitored in a university hospital between March 2017 and May 2018. In both the control and experimental groups, newborns were bathed and dried. However, in the control group, moisturizer was applied immediately to the newborn's body, whereas in the experimental group, moisturizer was applied 10 minutes after the completion of the bath. Researchers evaluated the BT and SM of all infants both before and immediately after the bath and at 10, 20, 40, and 60 minutes postbath. The control and experimental groups were similar according to the descriptive characteristics of the infants (P > .05). In both groups, infants' SM values increased in the first 10 minutes after the bath compared with the prebath values (P < .05). However, the whole-body SM value of the experimental group was significantly higher than that of the control group 60 minutes postbath (P = .027). There was also a statistically significant change in the body temperatures of infants in both groups after bathing (P = .004). Waiting 10 minutes postbath before applying moisturizer positively affected newborns' SM and BT. Additional research with a broader age range and a more diverse sample is needed to further clarify the effects of postbath moisturizer application timing on newborns' SM and BT.


Rates and symptoms of PTSD among cocaine-dependent patients.
This study evaluated lifetime traumatic events and current posttraumatic stress disorder (PTSD) symptoms in a substance abuse sample. Participants in the study consisted of 558 (75.1% male) cocaine-dependent individuals who completed self-report measures of trauma and PTSD symptoms prior to entry into treatment. Results showed a high number of lifetime traumatic events, even among those without PTSD. General disaster was the most prevalent. Current PTSD was found in 10.9% of the participants, with a significantly higher rate among women (21.6%) than among men (7.2%). For those with PTSD, the most prominent PTSD symptom cluster was arousal, and the most common symptoms were restricted affect, detachment and irritability. Participants with PTSD endorsed a large number of symptoms, almost double that needed to meet diagnostic criteria; however, neither number of traumas nor type of trauma was associated with their level of PTSD symptoms. Even among those not meeting PTSD criteria, subthreshold symptoms were found, with avoidance the most prominent cluster. Sociodemographic and recent cocaine use variables did not differentiate the PTSD from non-PTSD groups. PTSD is present in a sizeable percentage of cocaine-dependent treatment-seeking patients, particularly women. Clinicians might address arousal symptoms in particular, which were the most prominent symptom cluster, and which may be exacerbated by cocaine use. Even among those without PTSD, lifetime trauma is substantial and subthreshold PTSD symptoms are common. Vulnerability to PTSD needs further study, as sociodemographic and cocaine use variables did not distinguish between PTSD and non-PTSD groups.


Phage-associated mutator phenotype in group A streptococcus.
Defects in DNA mismatch repair (MMR) occur frequently in natural populations of pathogenic and commensal bacteria, resulting in a mutator phenotype. We identified a unique genetic element in Streptococcus pyogenes strain SF370 that controls MMR via a dynamic process of prophage excision and reintegration in response to growth. In S. pyogenes, mutS and mutL are organized on a polycistronic mRNA under control of a common promoter. Prophage SF370.4 is integrated between the two genes, blocking expression of the downstream gene (mutL) and resulting in a mutator phenotype. However, in rapidly growing cells the prophage excises and replicates as an episome, allowing mutL to be expressed. Excision of prophage SF370.4 and expression of MutL mRNA occur simultaneously during early logarithmic growth when cell densities are low; this brief window of MutL gene expression ends as the cell density increases. However, detectable amounts of MutL protein remain in the cell until the onset of stationary phase. Thus, MMR in S. pyogenes SF370 is functional in exponentially growing cells but defective when resources are limiting. The presence of a prophage integrated into the 5' end of mutL correlates with a mutator phenotype (10(-7) to 10(-8) mutation/generation, an approximately a 100-fold increase in the rate of spontaneous mutation compared with prophage-free strains [10(-9) to 10(-10) mutation/generation]). Such genetic elements may be common in S. pyogenes since 6 of 13 completed genomes have related prophages, and a survey of 100 strains found that about 20% of them are positive for phages occupying the SF370.4 attP site. The dynamic control of a major DNA repair system by a bacteriophage is a novel method for achieving the mutator phenotype and may allow the organism to respond rapidly to a changing environment while minimizing the risks associated with long-term hypermutability.


Identification of a novel genetic element in Escherichia coli K-12.
Induction of the SOS repair processes of Escherichia coli K-12 caused a 14.4-kilobase species of circular deoxyribonucleic acid, called element e14, to be excised from the chromosome. To aid further characterization of this species, an 11.6-kilobase segment of e14 was inserted into the HindIII site of plasmid pBR313. To map e14 on the E. coli K-12 chromosome, the recombinant plasmid, pAG2, was used to transform a polA recipient, an event which required integration of pAG2 into the recipient chromosome. This recombinational event was dependent upon the region of homology between the incoming plasmid and the chromosome, as no transformants were scored when either a strain cured of the element was the recipient or pBR313 was the transforming deoxyribonucleic acid. Using these transformants, we have shown that e14 maps between the purB and pyrC loci near min 25. Several strains of E. coli K-12 were found to contain e14; however, one strain, Ymel trpA36, did not. In addition, e14 was found to be absent in both E. coli B/5 and E. coli C. The approach to mapping developed for this work could be used to map other fragments of E. coli deoxyribonucleic acid which have no known phenotype.


Association of Engrailed homeoproteins with vesicles presenting caveolae-like properties.
We report here that the homeoproteins Engrailed-1 and Engrailed-2 are present in specific non-nuclear subcellular compartments. Using electron microscopy, we observed that chick-Engrailed-2 expressed in COS-7 cells associates with membrane fractions that are characterized as caveolae. This characterization is based on morphological, biochemical and immunological criteria such as, in particular, the absence of clathrin coat and the presence of caveolin and cholera toxin-binding sites. These data are fully confirmed by subcellular fractionation experiments, which demonstrate that transfected chick-Engrailed-2 is present in low density membrane fractions that are resistant to Triton X-100, enriched in caveolin and solubilized by the addition of a cholesterol-binding detergent, a set of properties highly characteristic of caveolae. The association of Engrailed-2 with specific membrane fractions observed after transfection in COS-7 cells is also observed for endogenous Engrailed-1 and Engrailed-2 expressed at late embryonic stages in the cerebellum and posterior mesencephalon of the rodent. Indeed, the two proteins are present in membrane fractions that bear all the characteristics of microdomains or caveolae-like domains, i.e. Triton X-100 resistance, saponin solubilization, low density on sucrose gradients, enrichment in glycosphingolipid GM1, absence of transmembrane Neural Cell Adhesion Molecule, presence of the glypiated (GPI-anchored) glycoprotein F3/F11 and of the acylated growth-associated protein GAP-43. Finally we demonstrate that part of the membrane-associated Engrailed, either expressed in COS-7 cells or endogenously present in neural tissues, is not accessible to proteolytic enzymes unless the membranes have been permeabilized with detergent. This study suggests that, in addition to their well-known presence in the nucleus, Engrailed proteins are also associated with caveolae-like vesicles that are primarily transported anterogradely into the axon, and that they can get access to a compartment compatible with secretion.


Healthcare system factors and colorectal cancer screening.
Developing effective programs to promote colorectal cancer (CRC) screening requires understanding of the effect of healthcare system factors on access to screening and adherence to guidelines. This study assessed the role of insurance status, type of plan, the frequency of preventive health visits, and provider recommendation on utilization of CRC screening tests using a cross-sectional, random-digit-dial survey of 1002 Massachusetts residents aged > or =50. A broad definition of CRC screening status included colonoscopy or barium enema (screening or diagnostic) within 10 years, flexible sigmoidoscopy (FSIG) within 5 years, and fecal occult blood testing (FOBT) in the past year as options; 51.7% of subjects aged 50 to 64 and 61.5% of older subjects were current. The uninsured had the lowest current testing rate. Among insured participants, type of insurance had little impact on CRC testing; older subjects enrolled in HMOs had marginally higher rates, although not statistically significant. Increased frequency of preventive health visits and ever receiving a physician's recommendation for FSIG or ever receiving FOBT cards were associated with higher rates of CRC screening among both age groups. Even when broad criteria are used to define current CRC screening status, a substantial proportion of the age-eligible population remains underscreened. Obtaining regular preventive care and receiving a physician's recommendation for screening appear to be potent facilitators of screening that should be considered in designing promotional efforts.


The physician at a distance: the use of videoconferencing in the treatment of patients with hypertension.
We evaluated the feasibility and quality of uncomplicated hypertension care based on telemedicine in a rural area of northern Sweden. The intervention subjects were 91 consecutive patients with primary hypertension. For comparison, 182 age- and sex-matched patients with hypertension were randomly selected from a similar health centre. The telemedicine equipment consisted of a video link between the physician and the patients, supported by a system for accessing medical data via the Internet. During a 21-month study period, telemedicine was used in 270 (91%) of the 297 consultations in the intervention group. All health personnel involved in the telemedicine treatment rated the method as feasible. Both systolic and diastolic blood pressure improved in the two groups during the study period. In the intervention group, a higher proportion had their blood pressure within treatment goals (systolic blood pressure <140 mmHg, diastolic blood pressure <90 mmHg) both at baseline and at follow-up than in the comparison group. An adjusted multivariate model (adjustment for sex, age, time between visits, change in number of drugs between first and last visit, blood pressure at first visit) showed that the intervention group had a higher probability (OR 2.7, 95% CI 1.4-5.2) of reaching the target blood pressure levels than the reference group. Treatment of hypertension by means of telemedicine was quite feasible and at least as effective as face-to-face consultations with a physician.


Addressing the challenges of a cross-national investigation: lessons from the Pittsburgh-Pisa study of treatment-relevant phenotypes of unipolar depression.
To date, no cross-national RCT has addressed the mechanisms underlying the relative success of pharmacological and psychotherapeutic interventions for depression. A multi-site clinical trial that includes psychotherapy as one of the treatments presents numerous challenges related to cross-site consistency and communication. This report describes how those challenges were met in the study "Depression: The Search for Treatment Relevant Phenotypes'', being carried out at the University of Pittsburgh and the University of Pisa, Italy. Implementing the study required the investigators to address methodological and practical challenges related to the different requirements of the two Institutional Review Boards (IRBs), psychotherapy training, independent evaluator training, patient recruitment, development of common tools for data entry, quality control and generation of weekly reports of patient progress as well as establishing a similar clinical and research framework in two countries with substantially different health care systems. By having bilingual investigators and staff members who spent time at one another's sites, making use of frequent conference-call staff meetings and being flexible within the bounds of the sometimes contradictory requirements of the IRBs, the investigators were able to meet the human subjects protection requirements of both institutions, surmount language barriers to consistent therapist and evaluator training and develop common tools for study management. As a result, recruitment goals were met at both sites and retention rates were high. One instance of inconsistent implementation of the protocol was corrected within the first year. This study was conducted in two Western cultures by researchers with long-standing collaboration. Our findings may not be generalizable to other countries or research settings. The implementation of a cross-national protocol and the adoption and maintenance of common procedures is possible when investigators are aware of the challenges this may present and are proactive in trying to address them.


In vivo protection against ZIKV infection and pathogenesis through passive antibody transfer and active immunisation with a prMEnv DNA vaccine.
Significant concerns have been raised owing to the rapid global spread of infection and disease caused by the mosquito-borne Zika virus (ZIKV). Recent studies suggest that ZIKV can also be transmitted sexually, further increasing the exposure risk for this virus. Associated with this spread is a dramatic increase in cases of microcephaly and additional congenital abnormalities in infants of ZIKV-infected mothers, as well as a rise in the occurrence of Guillain Barre' syndrome in infected adults. Importantly, there are no licensed therapies or vaccines against ZIKV infection. In this study, we generate and evaluate the in vivo efficacy of a novel, synthetic, DNA vaccine targeting the pre-membrane+envelope proteins (prME) of ZIKV. Following initial in vitro development and evaluation studies of the plasmid construct, mice and non-human primates were immunised with this prME DNA-based immunogen through electroporation-mediated enhanced DNA delivery. Vaccinated animals were found to generate antigen-specific cellular and humoral immunity and neutralisation activity. In mice lacking receptors for interferon (IFN)-α/β (designated IFNAR-/-) immunisation with this DNA vaccine induced, following in vivo viral challenge, 100% protection against infection-associated weight loss or death in addition to preventing viral pathology in brain tissue. In addition, passive transfer of non-human primate anti-ZIKV immune serum protected IFNAR-/- mice against subsequent viral challenge. This study in NHP and in a pathogenic mouse model supports the importance of immune responses targeting prME in ZIKV infection and suggests that additional research on this vaccine approach may have relevance for ZIKV control and disease prevention in humans.


Role of Env in resistance of feline immunodeficiency virus (FIV)-infected cats to superinfection by a second FIV strain as determined by using a chimeric virus.
A more or less pronounced resistance to superinfection by a second strain of the infecting virus has been observed in many lentivirus-infected hosts. We used a chimeric feline immunodeficiency virus (FIV), designated FIVchi, containing a large part of the env gene of a clade B virus (strain M2) and all the rest of the genome of a clade A virus (a p34TF10 molecular clone of the Petaluma strain modified to grow in lymphoid cells), to gain insights into such resistance. FIVchi was infectious and moderately pathogenic for cats and in vitro exhibited the neutralization specificity of the env donor. The experiments performed were bidirectional, in that cats preinfected with either parental virus were challenged with FIVchi and vice versa. The preinfected animals were partially or completely protected relative to what was observed in naïve control animals, most likely due, at least in part, to the circumstance that in all the preinfecting/challenge virus combinations examined, the first and the second virus shared significant viral components. Based on the proportions of complete protection observed, the role of a strongly matched viral envelope appeared to be modest and possibly dependent on the time interval between the first and the second infection. Furthermore, complete protection and the presence of measurable neutralizing antibodies capable of blocking the second virus in vitro were not associated.


MicroRNA-based silencing of Delta/Notch signaling promotes multiple cilia formation.
Multiciliated cells lining the surface of some vertebrate epithelia are essential for various physiological processes, such as airway cleansing. Their apical surface is constituted by hundreds of motile cilia, which beat in a coordinated manner to generate directional fluid flow. We recently reported the identification of microRNAs of the miR-449 family as evolutionary conserved key regulators of vertebrate multiciliogenesis. This novel function of miR-449 was established using in vivo and in vitro antisense approaches in two distinct experimental models. miR-449 strongly accumulated in multiciliated cells in human airway epithelium and Xenopus laevis embryonic epidermis, where it triggered centriole multiplication and multiciliogenesis by directly repressing the Delta/Notch pathway. Our data complement previous reports that showed the blocking action of miR-449 on the cell cycle, and unraveled a novel conserved mechanism whereby Notch signaling must undergo microRNA-mediated inhibition to permit differentiation of ciliated cell progenitors. We review here several important questions regarding the links between microRNAs and the Notch pathway in the control of cell fate.


[Behavioural attitude of the young as regards road accident prevention: a survey carried out among high school students in two Sicilian towns].
This survey is aimed at assessing young peoples awareness as regards car accident prevention and misleading behaviours. The survey concerned a sample of 829 high school students of both genders, aged 17 to 23, from a chief town (Catania) and a country town (Noto, Siracusa). In addition to a good level of awareness as regards road accidents and misleading behaviours, the investigation also showed an opposite trend to disregard basic safety rules such as the use of helmets, safety belts and abstention from alcohol intake. Dangerous behaviours have shown to be higher in Noto than Catania, and more frequent among male than female of both towns. In the light of such results, some educational actions have been discussed for road safety.


Gut Microbiota Promote Angiotensin II-Induced Arterial Hypertension and Vascular Dysfunction.
The gut microbiome is essential for physiological host responses and development of immune functions. The impact of gut microbiota on blood pressure and systemic vascular function, processes that are determined by immune cell function, is unknown. Unchallenged germ-free mice (GF) had a dampened systemic T helper cell type 1 skewing compared to conventionally raised (CONV-R) mice. Colonization of GF mice with regular gut microbiota induced lymphoid mRNA transcription of T-box expression in T cells and resulted in mild endothelial dysfunction. Compared to CONV-R mice, angiotensin II (AngII; 1 mg/kg per day for 7 days) infused GF mice showed reduced reactive oxygen species formation in the vasculature, attenuated vascular mRNA expression of monocyte chemoattractant protein 1 (MCP-1), inducible nitric oxide synthase (iNOS) and NADPH oxidase subunit Nox2, as well as a reduced upregulation of retinoic-acid receptor-related orphan receptor gamma t (Rorγt), the signature transcription factor for interleukin (IL)-17 synthesis. This resulted in an attenuated vascular leukocyte adhesion, less infiltration of Ly6G(+) neutrophils and Ly6C(+) monocytes into the aortic vessel wall, protection from kidney inflammation, as well as endothelial dysfunction and attenuation of blood pressure increase in response to AngII. Importantly, cardiac inflammation, fibrosis and systolic dysfunction were attenuated in GF mice, indicating systemic protection from cardiovascular inflammatory stress induced by AngII. Gut microbiota facilitate AngII-induced vascular dysfunction and hypertension, at least in part, by supporting an MCP-1/IL-17 driven vascular immune cell infiltration and inflammation.


Predictors of 30-Day Readmission After Inpatient Endoscopic Retrograde Cholangiopancreatography: A Single-Center Experience.
The 30-day hospital readmission rate is a nationally recognized quality measure. Nearly one-fifth of medicare beneficiaries are hospitalized within 30 days of discharge, resulting in a cost of over $26 billion dollars annually. Endoscopic retrograde cholangiopancreatography (ERCP) remains the endoscopic procedure with the highest risk of morbidity and mortality. We set out to analyze the clinical characteristics predictive of 30-day readmission after an inpatient ERCP. We performed a retrospective chart review of all inpatient ERCPs performed at our institution between 12/1/2014 and 9/30/2018. Clinical characteristics and outcomes of these patients were compared to determine predictors of 30-day readmission. A total of 497 inpatient ERCP procedures done for biliary or pancreatic indications, constituting 483 patients, were identified. There were 52 readmissions that occurred among 48 patients within 30 days of discharge. Basic demographic characteristics were similar between both groups. Comorbidities were significantly higher in those who were readmitted. Multivariate analysis revealed significantly greater odds of readmission with prior liver transplantation (OR = 4.15), cirrhosis (OR = 3.20), and pancreatic duct stent placement (OR = 2.56). Subgroup analysis for biliary indications revealed cholecystectomy before discharge and early ERCP to be protective against readmission. A history of liver transplantation and cirrhosis are predictive of increased 30-day readmission rates after an inpatient ERCP. Pancreatic duct stent placement is associated with readmission; however, this phenomenon is likely related to stenting for pancreatic endotherapy. Cholecystectomy before discharge and early ERCP are predictive of decreased need for readmission in procedures done for biliary indications.


A line, not a space, represents visual distinctness of borders formed by different colors.
When observers are asked to rate the visual distinctness of borders formed by the junction of two photic stimuli, normal trichromatic subjects behave in a manner similar to that of tritanopes in a color mixture experiment. All stimuli that look the same to the tritanope produce the same border distinctness with any other stimulus. Sets of such stimuli, whose members do not form borders with each other, map as single points along a curved line, where the Euclidean distance between pairs of points representing the two stimuli is nearly proportional to the rated distinctness of the border formed between them. In the absence of luminance differences, the perception of contour apparently depends on the stimulation of only two cone types.


Phylogenetic diversity and North Andean block conservation.
The Northern Andean Block (NAB) harbors high biodiversity; therefore, it is one of the most important areas in the Neotropics. Nevertheless, the settlement of several human populations has triggered the rapid transformation of ecosystems, leading to the extinction or endangerment of many species. Because phylogenetic diversity indices quantify the historical distinctness between species, they are adequate tools for evaluating priority conservation areas. We reconstructed 93 phylogenies encompassing 1,252 species and, utilizing their occurrence data sourced from the Global Biodiversity Information Facility, computed the Average Taxonomic Distinctness Index (AvTD) for each grid cell with a spatial resolution of 0.25° within the NAB. The index values for each grid cell were categorized into quantiles, and grid cells displaying values within the upper quantile (Q5) were identified as the most significant in terms of phylogenetic diversity. We also calculated the contribution of endemic species to overall phylogenetic diversity within the NAB, specifically focusing on areas preserved within protected areas. The NAB Andean region exhibited the highest AvTD, with high AvTD values observed in the middle and southern areas of Cordilleras. Endemic species made a relatively modest contribution to the overall phylogenetic diversity of the NAB, accounting for only 1.2% of the total. Despite their relatively small geographical footprints, protected areas within the NAB have emerged as crucial repositories of biodiversity, encompassing 40% of the total phylogenetic diversity in the region. Although the NAB Andean region has been identified as the most crucial area in terms of AvTD, some regions in the Amazonian Piedemonte and Pacific lowlands have high AvTD levels. Interestingly, some protected areas have been found to harbor higher AvTDs than expected, given their smaller size. Although the delimitation of new PAs and species richness have been the primary factors driving the expansion of PAs, it is also essential to consider the evolutionary information of species to conserve all aspects of biodiversity, or at least cover most of them. Therefore, using phylogenetic diversity measures and the results of this study can contribute to expanding the PA network and improving the connectivity between PAs. This approach will help conserve different aspects of biodiversity and preserve evolutionary relationships between species.


Role of pigment epithelium-derived factor in the reproductive system.
The physiological function of the female reproductive organs is hormonally controlled. In each cycle, the reproductive organs undergo tissue modifications that are accompanied by formation and destruction of blood vessels. Proper angiogenesis requires an accurate balance between stimulatory and inhibitory signals, provided by pro- and anti-angiogenic factors. As with many other tissues, vascular endothelial growth factor (VEGF) appears to be one of the major pro-angiogenic factors in the female reproductive organs. Pigment epithelium-derived factor (PEDF) is a non-inhibitory member of the serine protease inhibitors (serpin) superfamily, possessing potent physiologic anti-angiogenic activity that negates VEGF activity. The role of PEDF in decreasing abnormal neovascularization by exerting its anti-angiogenic effect that inhibits pro-angiogenic factors, including VEGF, has been investigated mainly in the eye and in cancer. This review summarizes the function of PEDF in the reproductive system, showing its hormonal regulation and its anti-angiogenic activity. Furthermore, some pathologies of the female reproductive organs, including endometriosis, ovarian hyperstimulation syndrome, polycystic ovary syndrome, and others, are associated with a faulty angiogenic process. This review illuminates the role of PEDF in their pathogenesis and treatment. Collectively, we can conclude that although PEDF seems to play an essential role in the physiology and pathophysiology of the reproductive system, its full role and mechanism of action still need to be elucidated.


The distribution of different molecular species of collagen in fibrous, elastic and hyaline cartilages of the pig.
The distribution of type II collagen, considered to be characteristic of cartilaginous tissues, was determined in various specialized cartilages of the mature pig. The tissues examined were: (1) fibrocartilage of the semilunar meniscus of the knee; (2) elastic cartilage of the external ear; (3) hyaline cartilage of (a) the synovial joint (b) the thyroid plate of the larynx, and (c) the nasal septum. The predominant species of collagen in each tissue, whether type I or type II, was appraised semi-quantitatively by analysis of purified collagen solubilized by pepsin and of peptide fragments produced by cyanogen bromide. Cyanogen bromide-derived peptides were characterized by column chromatography on CM-cellulose and by electrophoresis in sodium dodecyl sulphate-polyacrylamide gels. The proportion of each type of collagen was determined precisely by isolating the homologous small peptides alpha1(II)CB6 [nomenclature of Miller (1973) Clin. Orthop. 92, 260-280], by column chromatography on phosphocellulose and determining their relative proportions by amino acid analysis. Thus collagen of the fibrocartilage of the meniscus proved to be all type I; type II was not detected. In contrast, collagen of elastic cartilage of the outer ear, after rigorous exclusion of perichondrium, was type II. Similarly, type II was the only collagen detected in all the mature hyalline cartilages examined.


Customizing skills for assistive robotic manipulators, an inverse reinforcement learning approach with error-related potentials.
Robotic assistance via motorized robotic arm manipulators can be of valuable assistance to individuals with upper-limb motor disabilities. Brain-computer interfaces (BCI) offer an intuitive means to control such assistive robotic manipulators. However, BCI performance may vary due to the non-stationary nature of the electroencephalogram (EEG) signals. It, hence, cannot be used safely for controlling tasks where errors may be detrimental to the user. Avoiding obstacles is one such task. As there exist many techniques to avoid obstacles in robotics, we propose to give the control to the robot to avoid obstacles and to leave to the user the choice of the robot behavior to do so a matter of personal preference as some users may be more daring while others more careful. We enable the users to train the robot controller to adapt its way to approach obstacles relying on BCI that detects error-related potentials (ErrP), indicative of the user's error expectation of the robot's current strategy to meet their preferences. Gaussian process-based inverse reinforcement learning, in combination with the ErrP-BCI, infers the user's preference and updates the obstacle avoidance controller so as to generate personalized robot trajectories. We validate the approach in experiments with thirteen able-bodied subjects using a robotic arm that picks up, places and avoids real-life objects. Results show that the algorithm can learn user's preference and adapt the robot behavior rapidly using less than five demonstrations not necessarily optimal.


Comprehensive and quantitative analysis of lysophospholipid molecular species present in obese mouse liver by shotgun lipidomics.
Shotgun lipidomics exploits the unique chemical and physical properties of lipid classes and individual molecular species to facilitate the high-throughput analysis of a cellular lipidome on a large scale directly from the extracts of biological samples. A platform for comprehensive analysis of lysophospholipid (LPL) species based on shotgun lipidomics has not been established. Herein, after extensive characterization of the fragmentation pattern of individual LPL class and optimization of all experimental conditions including developing new methods for optimization of collision energy, and recovery and enrichment of LPL classes from the aqueous phase after solvent extraction, a new method for comprehensive and quantitative analysis of LPL species was developed. This newly developed method was applied for comprehensive analysis of LPL species present in mouse liver samples. Remarkably, the study revealed significant accumulation of LPL species in the liver of ob/ob mice. Taken together, by exploiting the principles of shotgun lipidomics in combination with a novel strategy of sample preparation, LPL species present in biological samples can be determined by the established method. We believe that this development is significant and useful for understanding the pathways of phospholipid metabolism and for elucidating the role of LPL species in signal transduction and other biological functions.


Diastereoselective Self-Assembly of Low-Symmetry Pdn L2n Nanocages through Coordination-Sphere Engineering.
Metal-organic cages (MOCs) are popular host architectures assembled from ligands and metal ions/nodes. Assembling structurally complex, low-symmetry MOCs with anisotropic cavities can be limited by the formation of statistical isomer libraries. We set out to investigate the use of primary coordination-sphere engineering (CSE) to bias isomer selectivity within homo- and heteroleptic Pdn L2n cages. Unexpected differences in selectivities between alternative donor groups led us to recognise the significant impact of the second coordination sphere on isomer stabilities. From this, molecular-level insight into the origins of selectivity between cis and trans diastereoisomers was gained, highlighting the importance of both host-guest and host-solvent interactions, in addition to ligand design. This detailed understanding allows precision engineering of low-symmetry MOC assemblies without wholesale redesign of the ligand framework, and fundamentally provides a theoretical scaffold for the development of stimuli-responsive, shape-shifting MOCs.


[Effects of polyvitamin deficiency on the activity of xenobiotic-metabolizing enzymes in rat liver].
The activity of xenobiotic-metabolizing enzymes was studied in the liver of male Wistar rats, which were fed for 4 weeks diets, containing 100 (control), 50 and 20% of vitamin adequate level. Moderate (50%) polyvitamin deficiency increased activity of EROD (by 13%), MROD (by 34%; p<0,05), 4-nitrophenol hydroxylase (by 16%), 6beta-testosterone hydroxylase (by 17%), UDP-glucuronosyle transferase (by 26%, p<0,05) and quinone reductase (by 55%, p<0,05). Deep (20%) polyvitamin deficiency decreased in liver activity of MROD (to 78% of control level, p<0,05), 4-nitrophenol hydroxylase (to 74%, p<0,05), heme oxygenase-1 (to 83%, p<0,05) and quinone reductase (to 60%, p<0,05). At the same time a 22% increase in the UDP-glucuronosyle transferase activity compared to the control group was found; activities of EROD, PROD, 6beta-testosterone hydroxylase and the total activity of glutathione S-transferase were unchanged. Deep polyvitamin deficiency had no significant effect on CYP1A1 mRNA and AhR mRNA level, whereas the expression of CYP1A2 mRNA and CYP3A1 mRNA were decreased to 62 and 79%, respectively, as compared with control.


Effect of glutamine on fructose 2,6-bisphosphate and on glucose metabolism in HeLa cells and in chick-embryo fibroblasts.
Glutamine caused a dose-dependent decrease in fructose 2,6-bisphosphate concentration in both HeLa cells and chick-embryo fibroblasts. The effect was complete within 15 min in HeLa cells, but required more than 9 h in the fibroblasts. Half-maximal effects were obtained with 0.1-0.3 mM-glutamine. In chick-embryo fibroblasts, but not in HeLa cells, glutamine induced a time-dependent decrease in the activity of phosphofructokinase-2, which correlated with the decrease in fructose 2,6-bisphosphate. Glutamine decreased the glycolytic flux by about 25% only in chick-embryo fibroblasts. The difference in glycolytic response between the two types of cells might correspond to a difference in the sensitivity of phosphofructokinase-1 for fructose 2,6-bisphosphate. In HeLa cells, glutamine caused a 2-3-fold stimulation of the synthesis of glycogen, a 50% decrease in the concentration of fructose 1,6-bisphosphate and a more than 80% decrease in the concentration of 5-phosphoribosyl pyrophosphate; the concentrations of hexose 6-phosphates and ATP were not affected.


Acute Steroid-Refractory Gastrointestinal Graft-Versus-Host Disease Is Not Associated With Significant Differences in Gut Taxonomic Composition Compared to Steroid-Sensitive Gastrointestinal Graft-Versus-Host Disease Immediately Before Onset of Disease.
Taxonomic composition of the gut microbiota at the time of neutrophil engraftment is associated with the development of acute gastrointestinal graft-versus-host disease (GI GVHD) in patients undergoing allogeneic hematopoietic stem cell transplantation. However, less is known about the relationship between the gut microbiota and development of steroid-refractory GI GVHD immediately before the onset of disease. Markers of steroid-refractory GI GVHD are needed to identify patients who may benefit from the early initiation of non-corticosteroid-based GVHD treatment. Our aim was to identify differences in taxonomic composition in stool samples from patients without GVHD, with steroid-responsive GVHD and with steroid-refractory GI GVHD to identify predictive microbiome biomarkers of steroid-refractory GI GVHD. We conducted a retrospective case-control, single institution study, performing shotgun metagenomic sequencing on stool samples from patients with (n = 36) and without GVHD (n = 34) matched for time since transplantation. We compared the taxonomic composition of the gut microbiome in those with steroid-sensitive GI GVHD (n = 17) and steroid-refractory GI GVHD (n = 19) to each other and to those without GVHD. We also performed associations between steroid-refractory GI GVHD, gut taxonomic composition, and fecal calprotectin, a marker of GI GVHD to develop composite fecal markers of steroid-refractory GVHD before the onset of GI disease. We found that fecal samples within 30 days of GVHD onset from patients with and without GVHD or with and without steroid-refractory GI GVHD did not differ significantly in Shannon diversity (alpha-diversity) or in overall taxonomic composition (beta-diversity). Although those patients without GVHD had higher relative abundance of Clostridium spp., those with and without steroid-refractory GI GVHD did not significantly differ in taxonomic composition between one another. In our study, fecal calprotectin before disease onset was significantly higher in patients with GVHD compared to those without GVHD and higher in patients with steroid-refractory GI GVHD compared to steroid-sensitive GI GVHD. No taxa were significantly associated with higher levels of calprotectin.


Experimental studies on the endothelium ultrastructure of heart capillaries under moderate (28-30 degrees) and deep (22-24 degrees) hypothermia without perfusion.
Ultrastructural changes in endothelial cells (EC) of myocardial capillaries were studied in 24 dogs which underwent hypothermia without perfusion. Biopsy specimens for electron microscopy were taken from the left ventricle of each dog in the control group, during anesthesia (prior to active cooling), and at the end of moderate (28-30 degrees ) and deep (22-24 degrees ) artificial body cooling. The following morphological types of the EC were identified both in the control group and in all test groups: those with moderately dense cytoplasm, light, dark, and irreversibly damaged cells. Dark cells showed increased numbers of plasmalemmal vesicles and appeared to be more transport-specialized as opposed to other types. In all stages of the experiment the amount of dark cells continuously increased (to 23.80, 34.62, and 47.17%, respectively). On cooling to 28-30 degrees, subcellular manifestation of reduced synthetic activity of organelles (nucleus, Golgi complex, and rough endoplasmic reticulum) was observed in all types of the EC. These changes persisted, or even increased, at the end of deep hypothermia. The transport activity of the EC changed differently in three experimental groups in all cell types. Micropinocytotic activity increased under spontaneous mild hypothermia (34-35 degrees ) during anesthesia and tended to decrease with subsequent artificial lowering of the temperature to 22-24 degrees. These ultrastructural changes seem to make up an integral part of the process of capillary endothelium adaptation to body surface cooling, and they might contribute to the development of tolerance to subsequent ischemic exposure during cardiac arrest.


Cellular therapies for spinal cord injury: what will the FDA need to approve moving from the laboratory to the human?
The transplantation of living cells and tissues to restore function and/or provide therapeutic molecules has been an active and ongoing area of research interest for over 25 years. Several of these potential therapies have reached initial clinical trials, and it is likely that applications will continue to expand, and that novel and improved approaches will be explored over the next several years. In the past, many of these experimental approaches were tested in early clinical trials without the oversight of regulatory agencies such as the Food and Drug Administration. However, as novel cellular therapies move from preclinical laboratory findings to the clinical arena, researchers and regulators face new and continually evolving issues and uncertainties involving long-term safety and efficacy. Using adrenal medullary transplantation in the spinal cord for pain as an example, this review presents an overview of past and current regulatory guidelines for moving these promising, novel cellular transplantation therapies from the laboratory to the human.


[Cytological method in the emergency surgical diagnosis of cancer at different sites].
The results of 1,215 emergency surgical cytological assays conducted in 619 cases of cancer and some other malignant tumors are presented. Specimens were obtained by puncture of primary tumor, swollen lymph nodes, indurations and infiltrations, by attrition of primary tumor, serous surfaces and surgical incisions, from serous cavity fluids and imprints of bioptical material. The results of emergency cytological diagnosis of cancer and other malignant tumors were confirmed in 95% of cases.