Low-Temperature Synchrotron Photoionization Study of 2-Methyl-3-buten-2-ol (MBO) Oxidation Initiated by O(3P) Atoms in the 298-650 K Range.
This work studies the oxidation of 2-methyl-3-buten-2-ol initiated by O(3P) atoms. The oxidation was investigated at room temperature, 550, and 650 K. Using the synchrotron radiation from the Advanced Light Source (ALS) of the Lawrence Berkley National Laboratory, reaction intermediates and products were studied by multiplexed photoionization mass spectrometry. Mass-to-charge ratios, kinetic time traces, photoionization spectra, and adiabatic ionization energies for each primary reaction species were obtained and used to characterize their identity. Using electronic structure calculations, potential energy surface scans of the different species produced throughout the oxidation were examined and presented in this paper to further validate the primary chemistry occurring. Branching fractions of primary products at all three temperatures were also provided. At room temperature only three primary products formed: ethenol (26.6%), acetaldehyde (4.2%), and acetone (53.4%). At 550 and 650 K the same primary products were observed in addition to propene (5.1%, 11.2%), ethenol (18.1%, 2.8%), acetaldehyde (8.9%, 5.7%), cyclobutene (1.6%, 10.8%), 1-butene (2.0%, 10.9%), trans-2-butene (3.2%, 23.1%), acetone (50.4%, 16.8%), 3-penten-2-one (1.0%, 11.5%), and 3-methyl-2-butenal (0.9%, 2.5%), where the first branching fraction value in parentheses corresponds to the 550 K data. At the highest temperature, a small amount of propyne (1.0%) was also observed.


Sphincter of Oddi manometry does not predispose to post-ERCP acute pancreatitis.
Sphincter of Oddi manometry is helpful in selecting patients with sphincter of Oddi dysfunction who will respond to sphincterotomy. However, studies have shown that sphincter of Oddi manometry is associated with a high risk of post-procedure pancreatitis. The primary objective of this study was to evaluate the safety of sphincter of Oddi manometry in patients with sphincter of 2Oddi dysfunction. The secondary objective was to determine the risk factors for post-ERCP pancreatitis in patients with sphincter of Oddi dysfunction. Data were collected retrospectively for 268 patients who had elective ERCP performed at 3 tertiary care medical centers between 1996 and 2000. Consecutive patients with suspected sphincter of Oddi dysfunction formed the case group; the control group consisted of patients with bile duct stone. The case group was further subclassified into group A, patients who underwent sphincter of Oddi manometry followed by immediate ERCP, and group B, patients who had ERCP without manometry. The rate of post-ERCP acute pancreatitis was compared between case and control groups. Twenty-seven percent of patients in the case group with suspected sphincter of Oddi dysfunction developed acute pancreatitis compared with 3.2% of patients in the control group with bile duct stone (p<0.001). There was no significant difference in the rate of acute pancreatitis in patients with sphincter of Oddi dysfunction who underwent sphincter of Oddi manometry and ERCP compared with patients with sphincter of Oddi dysfunction who had ERCP without sphincter of Oddi manometry (odds ratio 0.72: 95% CI[0.08, 9.2]). Multivariable logistic regression analysis showed that biliary sphincterotomy (p=0.006) and pancreatography (p=0.03) were independent predictors of acute pancreatitis. Patients with suspected sphincter of Oddi dysfunction are at higher risk of post-ERCP acute pancreatitis. Sphincter of Oddi manometry by itself does not appear to predispose to this complication.


In vitro evaluation of controlled-release 14% doxycycline gel for decontamination of machined and sandblasted acid-etched implants.
Peri-implant infections are associated with the establishment and maturation of a bacterial biofilm characterized by a predominance of Gram-negative fusiform anaerobic species. The decontamination of implant surfaces is then crucial for a successful peri-implant therapy. Twenty-one smooth and 21 rough implants, divided into four groups according to surface and treatment modality, were contaminated with Streptococcus sanguinis and then placed in an incubator with the atmosphere of 5% CO2 at 37°C for 24 hours to allow the bacteria to grow. After 24 hours, the test groups were treated with controlled release 14% doxycycline gel injecting the gel circumferentially over the surface of the implant for 3 minutes, while the control groups were irrigated with sterile saline for 1 minute. The implants were then vortexed into triptych soy broth to allow the bacteria to detach from the surface, diluted 1:100 and plated. Colony forming units (CFU) were counted 48 hours after incubation. The use of a 14% doxycycline gel minimized CFU counts compared to control groups, with the difference being statistically significant (P < 0.05). The reduction of CFUs in the smooth test group is more marked than in the rough test group, but the difference doesn't reach statistically significance (P = 0.215). The use of 14% doxycycline gel in implant surface decontamination was efficacious in this in-vitro study. Adjunctive use of locally delivered 14% doxycycline gel might be a viable option in the management of peri-implantitis and peri-implant mucositis considering its efficacy in reducing bacterial colonization.


Efficiency of ddRAD target enriched sequencing across spiny rock lobster species (Palinuridae: Jasus).
Double digest restriction site-associated DNA sequencing (ddRADseq) and target capture sequencing methods are used to explore population and phylogenetic questions in non-model organisms. ddRADseq offers a simple and reliable protocol for population genomic studies, however it can result in a large amount of missing data due to allelic dropout. Target capture sequencing offers an opportunity to increase sequencing coverage with little missing data and consistent orthologous loci across samples, although this approach has generally been applied to conserved markers for deeper evolutionary questions. Here, we combine both methods to generate high quality sequencing data for population genomic studies of all marine lobster species from the genus Jasus. We designed probes based on ddRADseq libraries of two lobster species (Jasus edwardsii and Sagmariasus verreauxi) and evaluated the captured sequencing data in five other Jasus species. We validated 4,465 polymorphic loci amongst these species using a cost effective sequencing protocol, of which 1,730 were recovered from all species, and 4,026 were present in at least three species. The method was also successfully applied to DNA samples obtained from museum specimens. This data will be further used to assess spatial-temporal genetic variation in Jasus species found in the Southern Hemisphere.


Analysis of sedentary behaviour levels and patterns in adults with Down syndrome.
Sedentary behaviour (SB) among adults with Down syndrome (DS) may differ based on personal or environmental factors. Investigate differences in SB levels and patterns of adults with DS based on sex, age, and residence-type across weekdays and weekends. Thirty-four adults with DS (15 men; 37 ± 12 years) underwent accelerometry-based measurements of sedentary time, bouts, and breaks for 7 days. We evaluated differences with 2 × 2 mixed-model (group-by-day) ANOVA. Younger (19-36 years) individuals had less sedentary time (p = .042), and shorter (p = .048) and fewer (p = .012) bouts than older (37-60 years) individuals. Group home residents had more bouts on weekends than adults living with parent/guardians (p = .015). Adults with DS spent half their waking hours in SB of short bouts. Age and residence may influence SB.


Deep brain stimulation compared with bariatric surgery for the treatment of morbid obesity: a decision analysis study.
Roux-en-Y gastric bypass is the gold standard treatment for morbid obesity, although failure rates may be high, particularly in patients with a BMI > 50 kg/m(2). With improved understanding of the neuropsychiatric basis of obesity, deep brain stimulation (DBS) offers a less invasive and reversible alternative to available surgical treatments. In this decision analysis, the authors determined the success rate at which DBS would be equivalent to the two most common bariatric surgeries. Medline searches were performed for studies of laparoscopic adjustable gastric banding (LAGB), laparoscopic Roux-en-Y gastric bypass (LRYGB), and DBS for movement disorders. Bariatric surgery was considered successful if postoperative excess weight loss exceeded 45% at 1-year follow-up. Using complication and success rates from the literature, the authors constructed a decision analysis model for treatment by LAGB, LRYGB, DBS, or no surgical treatment. A sensitivity analysis in which major parameters were systematically varied within their 95% CIs was used. Fifteen studies involving 3489 and 3306 cases of LAGB and LRYGB, respectively, and 45 studies involving 2937 cases treated with DBS were included. The operative successes were 0.30 (95% CI 0.247-0.358) for LAGB and 0.968 (95% CI 0.967-0.969) for LRYGB. Sensitivity analysis revealed utility of surgical complications in LRYGB, probability of surgical complications in DBS, and success rate of DBS as having the greatest influence on outcomes. At no values did LAGB result in superior outcomes compared with other treatments. Deep brain stimulation must achieve a success rate of 83% to be equivalent to bariatric surgery. This high-threshold success rate is probably due to the reported success rate of LRYGB, despite its higher complication rate (33.4%) compared with DBS (19.4%). The results support further research into the role of DBS for the treatment of obesity.


Changes in theta and alpha oscillatory signatures of attentional control in older and middle age.
Recent behavioural research has reported age-related changes in the costs of refocusing attention from a temporal (rapid serial visual presentation) to a spatial (visual search) task. Using magnetoencephalography, we have now compared the neural signatures of attention refocusing between three age groups (19-30, 40-49 and 60+ years) and found differences in task-related modulation and cortical localisation of alpha and theta oscillations. Efficient, faster refocusing in the youngest group compared to both middle age and older groups was reflected in parietal theta effects that were significantly reduced in the older groups. Residual parietal theta activity in older individuals was beneficial to attentional refocusing and could reflect preserved attention mechanisms. Slowed refocusing of attention, especially when a target required consolidation, in the older and middle-aged adults was accompanied by a posterior theta deficit and increased recruitment of frontal (middle-aged and older groups) and temporal (older group only) areas, demonstrating a posterior to anterior processing shift. Theta but not alpha modulation correlated with task performance, suggesting that older adults' stronger and more widely distributed alpha power modulation could reflect decreased neural precision or dedifferentiation but requires further investigation. Our results demonstrate that older adults present with different alpha and theta oscillatory signatures during attentional control, reflecting cognitive decline and, potentially, also different cognitive strategies in an attempt to compensate for decline.


Economic evaluations in water-fluoridation: a scoping review.
Community water fluoridation (CWF) is considered one of the 10 greatest public health achievements of the twentieth century and has been a cornerstone strategies for the prevention and control of dental caries in many countries. However, for decision-makers the effectiveness and safety of any given intervention is not always sufficient to decide on the best option. Economic evaluations (EE) provide key information that managers weigh, alongside other evidence. This study reviews the relevant literature on EE in CWF. A systematic database search up to August 2019 was carried out using MEDLINE, EMBASE, Cochrane Library, LILACS, Paediatric Economic Database Evaluation and National Health Service Economic Evaluation Database. The review included full economic evaluations on CWF programs, written in English, Spanish or Portuguese. The selection process and data extraction were carried out by two researchers independently. A qualitative synthesis of the results was performed. Of 498 identified articles, 24 studies met the inclusion criteria; 11 corresponded to cost-benefit analysis; nine were cost-effectiveness analyses; and four cost-utility studies. Two cost-utility studies used Disability-Adjusted Life Years,, one used Quality-Adjusted Tooth Years, and another Quality-Adjusted Life Years. EEs were conducted in eight countries. All studies concluded that water fluoridation was a cost-effective strategy when it was compared with non-fluoridated communities, independently of the perspective, time horizon or discount rate applied. Four studies adopted a lifetime time horizon. The outcome measures included caries averted (n = 14) and savings cost of dental treatment (n = 4). Most of the studies reported a caries reduction effects between 25 and 40%. Findings indicated that CWF represents an appropriate use of communities' resources, using a range of economic evaluation methods and in different locations. These findings provide evidence to decision-makers which they could use as an aid to deciding on resource allocation.


RVX-208, a stimulator of apolipoprotein AI gene expression for the treatment of cardiovascular diseases.
RVX-208 (RVX-000222) is a first-in-class, orally active, small-molecule stimulator of apolipoprotein (APO)AI gene expression, which is being developed by Resverlogix Corp for the potential treatment of cardiovascular diseases, in particular atherosclerosis and coronary artery disease. In vitro, RVX-208 stimulated APOAI transcription and was associated with dose-dependent increases in apoAI mRNA and protein. Toxicity studies in animals and phase I/II clinical trials have indicated that RVX-208 is safe and well tolerated in multiple dosing regimens. Plasma exposure to RVX-208 was dose-dependent following single or multiple oral doses and the drug was readily absorbed. In healthy volunteers and patients with low HDL-cholesterol levels, RVX-208 increased total HDL as well as the alpha- and pre-beta HDL fractions that are important substrates in the reverse cholesterol transport pathway. The results of further phase II trials are eagerly awaited to determine whether RVX-208 can deliver plaque regression via improvements in the plasma HDL profile of patients. In addition, a phase I trial indicated that RVX-208 may have potential for the removal of beta-amyloid plaques in Alzheimer's disease and this will be further assessed in an ongoing phase I/II clinical trial.


The development of a district community psychology service.
A survey was carried out to assess the difficulties encountered by general practitioners within a health district in the management of patients with emotional problems, and their attitudes to the involvement of clinical psychologists in primary health care.Based on the response to this survey a district community psychology service has been established. This is an adjunct to hospital-based services and attempts to overcome some of the shortcomings of traditional psychotherapeutic services.


Toward standardized amino acid matrices for exogenous phytase and protease in corn-soybean meal-based diets for broilers.
A total of 468 male Ross 308 broilers were used in a digestibility study to determine the additivity of apparent or standardized amino acid (AA) digestibility values for corn, soybean meal (SBM), or a mixture of corn and SBM that were supplemented, or not, with either phytase, protease, or a combination of phytase and protease. These treatments generated a total of 12 experimental diets that were arranged in a 3 × 4 design. A nitrogen-free diet was also fed to estimate endogenous AA loss. Apparent and standardized AA digestibility values were assessed on day 28 posthatch. The apparent digestibility of AA in the complete diet was higher (P < 0.05) than expected based on the digestibility of the corn and SBM individually. However, this overestimation was corrected by the adjustment to standardized values. Importantly, addition of protease or the combination of protease and phytase increased (P < 0.05) the digestibility of AA in corn and SBM. Furthermore, these effects were arithmetically coherent with respect to the measured effects of the enzymes in the mixture of corn and SBM, even improving the additivity of AA digestibility values when assessed on an apparent basis. This study demonstrates that the effect of exogenous protease and phytase on AA digestibility in complete diets is predictable based on measurements made in individual ingredients. In addition to improving digestibility values per se, exogenous protease and phytase may enhance precision in least cost formulation systems.


Guidelines for the Design and Conduct of Clinical Studies in Knee Articular Cartilage Repair: International Cartilage Repair Society Recommendations Based on Current Scientific Evidence and Standards of Clinical Care.
To summarize current clinical research practice and develop methodological standards for objective scientific evaluation of knee cartilage repair procedures and products. A comprehensive literature review was performed of high-level original studies providing information relevant for the design of clinical studies on articular cartilage repair in the knee. Analysis of cartilage repair publications and synopses of ongoing trials were used to identify important criteria for the design, reporting, and interpretation of studies in this field. Current literature reflects the methodological limitations of the scientific evidence available for articular cartilage repair. However, clinical trial databases of ongoing trials document a trend suggesting improved study designs and clinical evaluation methodology. Based on the current scientific information and standards of clinical care, detailed methodological recommendations were developed for the statistical study design, patient recruitment, control group considerations, study endpoint definition, documentation of results, use of validated patient-reported outcome instruments, and inclusion and exclusion criteria for the design and conduct of scientifically sound cartilage repair study protocols. A consensus statement among the International Cartilage Repair Society (ICRS) and contributing authors experienced in clinical trial design and implementation was achieved. High-quality clinical research methodology is critical for the optimal evaluation of current and new cartilage repair technologies. In addition to generally applicable principles for orthopedic study design, specific criteria and considerations apply to cartilage repair studies. Systematic application of these criteria and considerations can facilitate study designs that are scientifically rigorous, ethical, practical, and appropriate for the question(s) being addressed in any given cartilage repair research project.


Engineered Mesenchymal Stem Cells for Targeting Solid Tumors: Therapeutic Potential beyond Regenerative Therapy.
Mesenchymal stem cells (MSCs) have previously demonstrated considerable promise in regenerative medicine based on their ability to proliferate and differentiate into cells of different lineages. More recently, there has been a significant interest in using MSCs as cellular vehicles for targeted cancer therapy by exploiting their tumor homing properties. Initial studies focused on using genetically modified MSCs for targeted delivery of various proapoptotic, antiangiogenic, and therapeutic proteins to a wide variety of tumors. However, their use as drug delivery vehicles has been limited by poor drug load capacity. This review discusses various strategies for the nongenetic modification of MSCs that allows their use in tumor-targeted delivery of small molecule chemotherapeutic agents. SIGNIFICANCE STATEMENT: There has been considerable interest in exploiting the tumor homing potential of MSCs to develop them as a vehicle for the targeted delivery of cytotoxic agents to tumor tissue. The inherent tumor-tropic and drug-resistant properties make MSCs ideal carriers for toxic payload. While significant progress has been made in the area of the genetic modification of MSCs, studies focused on identification of molecular mechanisms that contribute to the tumor tropism along with optimization of the engineering conditions can further improve their effectiveness as drug delivery vehicles.


Recording of multiple holograms in photopolymer films.
Multiple images were recorded in DuPont HRF-150 photopolymer as transmission holograms by using angle multiplexing. Agreement with Kogelnik's two-wave diffraction theory for phase gratings, the time considerations and its effect on dynamic range, the necessity of a pre-illumination pulse, and good image quality at readout are reported.


Opsonizing activities of IgG, IgM antibodies and the C3b inactivator-cleaved third component of complement in macrophage phagocytosis.
Phagocytosis of SRBC by guinea-pig peritoneal macrophages is enhanced by opsonizing IgG antibody alone. IgM antibody requires the presence of bound C3. Treatment of C3b coated SRBC with purified C3b inactivator (yielding EAIgM C1423d) does not reduce attachment to, and phagocytosis by, peritoneal macrophages. This finding suggests the existence of a C3d receptor on peritoneal macrophages. EC43b intermediates which have been produced by removing IgM antibody by mercaptoethanol treatment and by subsequent removal of C1 and C2, are phagocytosed despite the absence of IgM antibody. Furthermore, treatment of EC43b with C3b inactivator does not change phagocytosis. Thus, IgM antibody does not appear to be a necessary prerequisite for the stimulation of phagocytosis, C3b or C3d alone being sufficient.


Co-assembled supramolecular hydrogels of cell adhesive peptide and alginate for rapid hemostasis and efficacious wound healing.
Injectable hydrogels are promising materials for applications in non-compressive wound management. Yet difficulties remain for the fabrication of mechanically stable hydrogel materials with inherent functionalities in both hemostatic control and wound healing without additional supplements of growth factors. Herein, we reported the co-assembly of a cell adhesive peptide conjugate (Pept-1) and alginate (ALG), to confer supramolecular hydrogels with excellent mechanical properties and high efficacy in both hemostatic control and wound healing requiring no additional growth factors. The co-assembling process of Pept-1 and ALG, which was mediated by electrostatic interactions and metal chelation, afforded a composite hydrogel with denser nanofibrillar structures and better mechanical strength when comparing to the Pept-1 gel alone. As-prepared Pept-1/ALG hydrogels exhibited excellent injectability and thixotropic properties, making them ideal materials for wound dressing. The composite hydrogel induced fast hemostasis when spiked with whole blood in vitro, and reduced the amount of bleeding to ∼18% of the untreated control in a liver puncture mouse model. Meanwhile, it promoted adhesion and migration of fibroblast NIH3T3 cells in vitro, and accelerated the rate of wound healing in a full-thickness skin defect model of mice. In addition, the Pept-1/ALG hydrogel showed excellent biocompatibility with no obvious hemolytic activity. In future, the strategy of utilizing co-assembled nanostructures composed of biofunctional peptides and polysaccharides could be further exploited to construct a broad range of nanocomposite materials for a variety of biomedical applications.


Coagulase and Efb of Staphylococcus aureus Have a Common Fibrinogen Binding Motif.
Coagulase (Coa) and Efb, secreted Staphylococcus aureus proteins, are important virulence factors in staphylococcal infections. Coa interacts with fibrinogen (Fg) and induces the formation of fibrin(ogen) clots through activation of prothrombin. Efb attracts Fg to the bacterial surface and forms a shield to protect the bacteria from phagocytic clearance. This communication describes the use of an array of synthetic peptides to identify variants of a linear Fg binding motif present in Coa and Efb which are responsible for the Fg binding activities of these proteins. This motif represents the first Fg binding motif identified for any microbial protein. We initially located the Fg binding sites to Coa's C-terminal disordered segment containing tandem repeats by using recombinant fragments of Coa in enzyme-linked immunosorbent assay-type binding experiments. Sequence analyses revealed that this Coa region contained shorter segments with sequences similar to the Fg binding segments in Efb. An alanine scanning approach allowed us to identify the residues in Coa and Efb that are critical for Fg binding and to define the Fg binding motifs in the two proteins. In these motifs, the residues required for Fg binding are largely conserved, and they therefore constitute variants of a common Fg binding motif which binds to Fg with high affinity. Defining a specific motif also allowed us to identify a functional Fg binding register for the Coa repeats that is different from the repeat unit previously proposed. Staphylococcus aureus infections are a major health problem that affects an estimated 50 million people globally and causes the death of about 20,000 Americans each year. A number of experimental vaccines have been developed during the past years. However, these vaccines have all failed in clinical trials. The ability of S. aureus to form an Fg shield surrounding and protecting bacterial cells from clearance may explain why the vaccines are failing. Furthermore, S. aureus coagulase can induce the formation of a fibrin(ogen) shield in experimental abscess models which surrounds and protects bacteria in the microcolony from clearance. In this study, we identified for the first time a microbial Fg binding motif. Variants of this motif are present in coagulase and Efb. Our results provide a molecular basis for the rational design of inhibitors that could potentially prevent the formation of the obstructing Fg shield.


Tertiary structure of an RNA pseudoknot is stabilized by "diffuse" Mg2+ ions.
The aim of this study is to obtain a comprehensive experimental and theoretical description of the contributions of Mg2+ ions to the free energy of folding a pseudoknot RNA tertiary structure. A fluorescence method for measuring the effective concentration of Mg2+ in the presence of an RNA was used to study Mg2+-RNA interactions with both folded and partially unfolded forms of an RNA pseudoknot. These data established the excess number of Mg2+ ions accumulated by the folded or partially unfolded RNAs as a function of bulk Mg2+ concentration, from which free energies of Mg2+-RNA interactions were derived. Complementary thermal melting experiments were also used to obtain RNA-folding free energies. These experimental data were compared with the results of calculations based on the nonlinear Poisson-Boltzmann equation, which describes the interaction of "diffuse" (fully hydrated) Mg2+ ions with the different RNA forms. Good agreement between the calculations and experimental data suggests that essentially all of the Mg2+-induced stabilization of the native pseudoknot structure arises from the stronger interaction of diffuse ions with the folded tertiary structure compared to that with a partially unfolded state. It is unlikely that the stability of the RNA depends on dehydrated ions bound to specific sets of RNA ligands in the folded state. The data also suggest that the Mg2+-dependent free energy of folding is sensitive to factors that influence the ensemble of RNA conformations present in the partially unfolded state.


Lipid compositions of intracellular membranes isolated from rat liver nodules in Wistar rats.
The mevalonate pathway gives rise to important end products for the regulation of growth and resistance to oxidative stress and is, consequently, of importance in carcinogenesis. In this study liver nodules were produced in Wistar rats by intermittent feeding with dietary 2-acetylaminofluorene, and the lipid compositions of isolated microsomes, mitochondria, and lysosomes were examined. The phospholipid compositions of these subfractions were unchanged compared to normal hepatic tissue, but the fatty acid patterns were altered, particularly in microsomes. An increase in the content of palmitic acid and a decrease in that of stearic acid were noted. The pattern of fatty acyl moieties on carbon atoms 1 and 2 of the glycerol backbone of phospholipids was unchanged in nodular tissue compared to normal liver. The amount of dolichol was significantly higher in microsomes and mitochondria, but not in lysosomes, and the relative amounts of longer polyisoprenoid compounds were increased in the liver nodules. The relative concentration of esterified dolichol was decreased and an enrichment in saturated fatty acids in this fraction could be observed. The cholesterol concentration was found to be lower in microsomes, but was unchanged in mitochondria and lysosomes, and the normally low concentration of cholesteryl esters was elevated somewhat in microsomes and lysosomes. The ubiquinone content of liver nodular mitochondria was unchanged, but increased 7-fold in microsomes and 2-fold in lysosomes. The alterations found in the lipid composition of liver nodules are significant and have functional implications in many cellular processes of proposed importance for the carcinogenic process, i.e., protein glycosylation cholesterogenesis, regulation of the mevalonate pathway, cellular oxidation-reduction state, and resistance to oxidative stress.


Higher viral load and infectivity increase risk of aerosol transmission for Delta and Omicron variants of SARS-CoV-2.
Airborne transmission of SARS-CoV-2 is an important route of infection. For the wildtype (WT) only a small proportion of those infected emitted large quantities of the virus. The currently prevalent variants of concern, Delta (B1.617.2) and Omicron (B.1.1.529), are characterized by higher viral loads and a lower minimal infective dose compared to the WT. We aimed to describe the resulting distribution of airborne viral emissions and to reassess the risk estimates for public settings given the higher viral load and infectivity. We reran the Monte Carlo modelling to estimate viral emissions in the fine aerosol size range using available viral load data. We also updated our tool to simulate indoor airborne transmission of SARS-CoV-2 by including a CO2 calculator and recirculating air cleaning devices. We also assessed the consequences of the lower critical dose on the infection risk in public settings with different protection strategies. Our modelling suggests that a much larger proportion of individuals infected with the new variants are high, very high or super-emitters of airborne viruses: for the WT, one in 1,000 infected was a super-emitter; for Delta one in 30; and for Omicron one in 20 or one in 10, depending on the viral load estimate used. Testing of the effectiveness of protective strategies in view of the lower critical dose suggests that surgical masks are no longer sufficient in most public settings, while correctly fitted FFP2 respirators still provide sufficient protection, except in high aerosol producing situations such as singing or shouting. From an aerosol transmission perspective, the shift towards a larger proportion of very high emitting individuals, together with the strongly reduced critical dose, seem to be two important drivers of the aerosol risk, and are likely contributing to the observed rapid spread of the Delta and Omicron variants of concern. Reducing contacts, always wearing well-fitted FFP2 respirators when indoors, using ventilation and other methods to reduce airborne virus concentrations, and avoiding situations with loud voices seem critical to limiting these latest waves of the COVID-19 pandemic.


Effects of a single dose of vitamin D in septic children: a randomized, double-blinded, controlled trial.
To assess the effects of a single dose of vitamin D on 25-hydroxyvitamin D (25OHD) levels and clinical outcomes in children with vitamin D deficiency (VDD) and sepsis. In this randomized, controlled trial, eligible children with VDD and sepsis were assigned to receive one dose of 150,000 IU of cholecalciferol or placebo. Serum concentrations of 25OHD, angiotensin-II (Ang-II), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were assessed at baseline and 8 days after treatment. The cardiovascular Sequential Organ Failure Assessment (cv-SOFA) score, septic shock incidence, duration of ventilation, and mortality were also examined. One hundred nine participants fulfilled the study requirements. The two groups had comparable baseline characteristics. Ang-II, IL-6, and TNF-α concentrations were all reduced after vitamin D supplementation. Furthermore, the cv-SOFA score (1.76 ± 0.8 vs. 2.3 ± 1.1) and incidence of septic shock (7% vs. 20%) were lower in the treatment group than in the control group. The duration of ventilation and mortality rates did not differ between two groups. A single dose of vitamin D improved 25OHD levels and the incidence of septic shock in children with VDD and sepsis.


Neurotoxins from marine dinoflagellates: a brief review.
Dinoflagellates are not only important marine primary producers and grazers, but also the major causative agents of harmful algal blooms. It has been reported that many dinoflagellate species can produce various natural toxins. These toxins can be extremely toxic and many of them are effective at far lower dosages than conventional chemical agents. Consumption of seafood contaminated by algal toxins results in various seafood poisoning syndromes: paralytic shellfish poisoning (PSP), neurotoxic shellfish poisoning (NSP), amnesic shellfish poisoning (ASP), diarrheic shellfish poisoning (DSP), ciguatera fish poisoning (CFP) and azaspiracid shellfish poisoning (ASP). Most of these poisonings are caused by neurotoxins which present themselves with highly specific effects on the nervous system of animals, including humans, by interfering with nerve impulse transmission. Neurotoxins are a varied group of compounds, both chemically and pharmacologically. They vary in both chemical structure and mechanism of action, and produce very distinct biological effects, which provides a potential application of these toxins in pharmacology and toxicology. This review summarizes the origin, structure and clinical symptoms of PSP, NSP, CFP, AZP, yessotoxin and palytoxin produced by marine dinoflagellates, as well as their molecular mechanisms of action on voltage-gated ion channels.


A supramolecular nanovehicle toward systematic, targeted cancer and tumor therapy.
A supramolecular nanovehicle (denoted as SNV) was fabricated by encapsulating zinc phthalocyanine (ZnPc) and doxorubicin (DOX) into a copolymer (PVP-b-PAA-g-FA), so as to achieve systematic and synergistic chemotherapy-photodynamic therapy (PDT), targeted tumor imaging and therapy. The sophisticated copolymer designed in this work can load the PDT photosensitizer (ZnPc) and chemotherapy drug (DOX) simultaneously, which exhibits an excellent performance in chemotherapy-PDT targeted cancer and tumor therapy for both in vitro studies performed with HepG2 cells and in vivo tests with mice. This work provides a new drug formulation with a chemotherapy-PDT synergistic effect by virtue of the supramolecular material design, which possesses the advantages of an ultra-low drug dosage and highly-efficient in vivo targeted tumor imaging/therapy.


Asymmetric Synthesis of Griffipavixanthone Employing a Chiral Phosphoric Acid-Catalyzed Cycloaddition.
Asymmetric synthesis of the biologically active xanthone dimer griffipavixanthone is reported along with its absolute stereochemistry determination. Synthesis of the natural product is accomplished via dimerization of a p-quinone methide using a chiral phosphoric acid catalyst to afford a protected precursor in excellent diastereo- and enantioselectivity. Mechanistic studies, including an unbiased computational investigation of chiral ion-pairs using parallel tempering, were performed in order to probe the mode of asymmetric induction.


Production of antikeratin autoantibodies by hybrid spleen cells of naive mice.
The mechanism of the occurrence of natural antikeratin antibodies in human sera was studied using hybrid spleen cells obtained from experimentally naive or from immunized mice. Antikeratin antibodies were detected by enzyme-linked immunosorbent assay (ELISA) in 5.9-9.5% of the culture supernatants of fused spleen cells taken from naive mice. When mice were immunized with keratins, the number of supernatants containing antikeratin antibodies was increased to eight out of 51 (15.7%). When immunized with non-keratin materials such as activated human T cells, adult T-cell leukaemia cell lysates, and human T-cell lymphotropic virus type-I (HTLV-I), 16.7-20.8% of the supernatants were found to contain antikeratin antibodies by ELISA. The antikeratin antibodies in the supernatants showed cytoplasmic staining of keratinocytes in human as well as mouse skin by indirect immunofluorescence. The antibodies reacted with extracted human epidermal keratins by dot-blot and Western blot analysis. Most antikeratin antibodies in the supernatants did not show cross-reactivity with exogenous antigens used for immunization and vimentin-type intermediate-sized filaments. These findings demonstrate that B cells producing antikeratin antibodies are common in naive mice, and produce various types of antikeratin antibodies following specific activation with epidermal keratins and non-specific immunological stimuli.


The effect of low molecular weight heparin on survival in patients with advanced malignancy.
Studies in cancer patients with venous thromboembolism suggested that low molecular weight heparin may prolong survival. In a double-blind study, we evaluated the effect of low molecular weight heparin on survival in patients with advanced malignancy without venous thromboembolism. Patients with metastasized or locally advanced solid tumors were randomly assigned to receive a 6-week course of subcutaneous nadroparin or placebo. The primary efficacy analysis was based on time from random assignment to death. The primary safety outcome was major bleeding. In total, 148 patients were allocated to nadroparin and 154 patients were allocated to placebo. Mean follow-up was 1 year. In the intention-to-treat analysis the overall hazard ratio of mortality was 0.75 (95% CI, 0.59 to 0.96) with a median survival of 8.0 months in the nadroparin recipients versus 6.6 months in the placebo group. After adjustment for potential confounders, the treatment effect remained statistically significant. Major bleeding occurred in five (3%) of nadroparin-treated patients and in one (1%) of the placebo recipients (P = .12). In the a priori specified subgroup of patients with a life expectancy of 6 months or more at enrollment, the hazard ratio was 0.64 (95% CI, 0.45 to 0.90) with a median survival of 15.4 and 9.4 months, respectively. For patients with a shorter life expectancy, the hazard ratio was 0.88 (95% CI, 0.62 to 1.25). A brief course of subcutaneous low molecular weight heparin favorably influences the survival in patients with advanced malignancy and deserves additional clinical evaluation.


The XylR variant (R121C and P363S) releases arabinose-induced catabolite repression on xylose fermentation and enhances coutilization of lignocellulosic sugar mixtures.
Microbial production of fuels and chemicals from lignocellulosic biomass provides a promising alternative to conventional petroleum-derived routes. However, the heterogeneous sugar composition of lignocellulose prevents efficient microbial sugar co-fermentation due to carbon catabolite repression, which negatively affects production metrics. We previously discovered that a mutant copy of the transcriptional regulator XylR (P363S and R121C; denoted as XylR*) in Escherichia coli has a higher DNA-binding affinity than wild-type XylR, leading to a stronger activation of the d-xylose catabolic genes and a release from glucose-induced repression on xylose fermentation. Here, we showed that XylR* also releases l-arabinose-induced repression on xylose fermentation through altered transcriptional control, enhancing co-fermentation of arabinose-xylose sugar mixtures in wild-type E. coli. Integrating xylR* into an ethanologenic E. coli resulted in the coutilization of 96% of the provided glucose-xylose-arabinose mixtures (120 g/L total sugars supplied) with an ethanol yield higher than 90% of the theoretical maximum by simple batch fermentations.


Adolescent victimization and associated suicidal and violent behaviors.
This study examined the relationship between victimization and both suicidal and violent behaviors among 1,569 public high school students in New York State (excluding New York City). They had participated in the 1997 Youth Risk Behavior Survey and, based on their responses, were divided into four groups: no suicidal or violent behavior, suicidal behavior only, violent behavior only, and both suicidal and violent behaviors. Males reported significantly more victimization, less suicidal behavior, and more violent behavior compared with females. Logistic regression analyses indicated that all categories of suicidal/violent behaviors were more frequent among those who had been victimized compared with the nonvictimized, for both males and females. In addition, victimized males were over two times more likely than victimized females to report violent behavior only. Gender differences were not significant for victimized students in two groups, suicidal behavior only and both suicidal and violent behaviors, although the results were in the hypothesized direction for suicidal behavior (e.g., females were more prone to suicidal behavior only). It was concluded that identifying and treating the victims of violence should be an integral component of suicide prevention programs and interventions aimed at reducing interpersonal violence in schools.


Transoral robotic surgery and a paradigm shift in the management of oropharyngeal squamous cell carcinoma.
Efforts to improve upon oncologic results for resectable oropharyngeal carcinoma over the past 40 years have yielded many treatment options. However, the two fundamental approaches, surgical and nonsurgical, have not been compared in a randomized trial. Both approaches can be effective and both can cause treatment-related swallowing complications. Early experience combining technological advancements with conformal radiotherapy and transoral robotic surgery suggests that the risk of swallowing complications may be reduced and that treatment outcomes may be improved through pathologic stratification and risk-adapted adjuvant therapy. This may also be a valid strategy upon which treatment de-intensification may be studied for human-papillomavirus-associated oropharyngeal carcinoma due to its favorable prognosis.


Effects of ACTH, dexamethasone, and adrenalectomy on 11beta-hydroxylase (CYP11B1) and aldosterone synthase (CYP11B2) gene expression in the rat central nervous system.
Using a highly sensitive quantitative RT-PCR method for the measurement of CYP11B1 (11beta-hydroxylase) and CYP11B2 (aldosterone synthase) mRNAs, we previously demonstrated that CYP11B2 expression in the central nervous system (CNS) is subject to regulation by dietary sodium. We have now quantified the expression of these genes in the CNS of male Wistar Kyoto (WKY) rats in response to systemic ACTH infusion, dexamethasone infusion, and to adrenalectomy. CYP11B1 and CYP11B2 mRNA levels were measured in total RNA isolated from the adrenal gland and discrete brain regions using real-time quantitative RT-PCR. ACTH infusion (40 ng/day for 7 days, N=8) significantly increased CYP11B1 mRNA in the adrenal gland, hypothalamus, and cerebral cortex compared with animals infused with vehicle only. ACTH infusion decreased adrenal CYP11B2 expression but increased expression in all of the CNS regions except the cortex. Dexamethasone (10 microg/day for 7 days, N=8) reduced adrenal CYP11B1 mRNA compared with control animals but had no significant effect on either gene's expression in the CNS. Adrenalectomy (N=6 per group) significantly increased CYP11B1 expression in the hippocampus and hypothalamus and raised CYP11B2 expression in the cerebellum relative to sham-operated animals. This study confirms the transcription of CYP11B1 and CYP11B2 throughout the CNS and demonstrates that gene transcription is subject to differential regulation by ACTH and circulating corticosteroid levels.


Noise-induced bistability in the quasi-neutral coexistence of viral RNAs under different replication modes.
Evolutionary and dynamical investigations into real viral populations indicate that RNA replication can range between the two extremes represented by so-called 'stamping machine replication' (SMR) and 'geometric replication' (GR). The impact of asymmetries in replication for single-stranded (+) sense RNA viruses has been mainly studied with deterministic models. However, viral replication should be better described by including stochasticity, as the cell infection process is typically initiated with a very small number of RNA macromolecules, and thus largely influenced by intrinsic noise. Under appropriate conditions, deterministic theoretical descriptions of viral RNA replication predict a quasi-neutral coexistence scenario, with a line of fixed points involving different strands' equilibrium ratios depending on the initial conditions. Recent research into the quasi-neutral coexistence in two competing populations reveals that stochastic fluctuations fundamentally alter the mean-field scenario, and one of the two species outcompetes the other. In this article, we study this phenomenon for viral RNA replication modes by means of stochastic simulations and a diffusion approximation. Our results reveal that noise has a strong impact on the amplification of viral RNAs, also causing the emergence of noise-induced bistability. We provide analytical criteria for the dominance of (+) sense strands depending on the initial populations on the line of equilibria, which are in agreement with direct stochastic simulation results. The biological implications of this noise-driven mechanism are discussed within the framework of the evolutionary dynamics of RNA viruses with different modes of replication.


U.S. police rosters: Fatigue and public complaints.
Recent experimental research indicates a substantial impact of shift work related fatigue on police officers' encounters with the public. In recent years, biomathematical models of fatigue have provided a new way to identify potential relationships between working time arrangements and job performance. This study focused on public complaints against police officers and determined whether the odds of a public complaint were associated with work schedules and/or a biomathematical model's predictions of fatigue and sleepiness. N = 144 police officers from two municipal police departments in the United States reported shift start times, shift hours, court hours, and public complaints each duty day during study participation. A biomathematical model of fatigue (FAID Quantum) predicted sleep duration and sleep timing and inferred fatigue and sleepiness for 15 744 shifts. Fatigue, sleepiness, 24 hr sleep estimates, and work schedule were tested as predictors of public complaints. Greater fatigue, greater sleepiness, and less sleep in the 24 hr prior to a shift increased the odds of a public complaint (F ≥ 9.14, p < 0.01). Working back-to-back night shifts increased the odds of a public complaint (OR = 4.27, p < 0.01), particularly when off-duty court hours were worked between the night shifts (OR = 4.73, p < 0.01). On-duty fatigue and sleepiness, sleep obtained prior to a shift, and working night shifts were strongly associated with public complaints. Off-duty court appearances reduced sleep between night shifts and further increased the odds of a public complaint. The results suggest that off-duty court hours should be limited between night shifts and duty schedules should be considered when scheduling court appearances.


Pathology of bursae of Fabricius in methionine-deficient broiler chickens.
The purpose of this 42-day study was to investigate the effects of methionine (Met) deficiency on immune function by determining the relative weight, morphological and ultrastructural changes of bursae of Fabricius, cell cycle, and apoptosis of bursa cells. One hundred and twenty one-day-old avian broilers were randomly divided into two groups and fed on a control diet (starter diet, Met 0.50%; grower diet, Met 0.40%) and Met-deficient diet (starter diet, Met 0.26%; grower diet, Met 0.28%) for six weeks. The relative weight of bursae was decreased with Met deficiency when compared to that of the control group. Lesions were also observed in the Met-deficient group. Histopathologically, the numbers of lymphocytes in the follicles were decreased. Ultrastructurally, the mitochondria of lymphocytes were swollen in the Met-deficient group. As measured by flow cytometry, bursal cells in the G0G1 phase were significantly higher (P < 0.01), and bursal cells in the S, G2M phases and proliferating index were obviously lower (P < 0.01) with Met deficiency than in the control group. Moreover, the percentage of apoptotic cells in the bursae were significantly increased in Met-deficient birds (P < 0.01). It was concluded that Met deficiency restrained the development of the bursae of Fabricius and affected the humoral immunity of the chickens.


Genome differentiation in Aegilops. 1. Distribution of highly repetitive DNA sequences on chromosomes of diploid species.
Genome differentiation in 12 diploid Aegilops species was analyzed using in situ hybridization with the highly repetitive DNA sequences pSc119 and pAs1 and C-banding. Chromosomes of all these diploid Aegilops species hybridized with the pSc119 probe; however, the level of hybridization and labeling patterns differed among genomes. Only four species (Ae. squarrosa, Ae. comosa, Ae. heldreichii, and Ae. uniaristata) showed distinct hybridization with pAs1. The labeling patterns were species-specific and chromosome-specific. Differences in in situ hybridization (ISH) patterns, also observed by C-banding, exist between the karyotypes of Ae. comosa and Ae. heldreichii, suggesting that they are separate, although closely related, subspecies. The S genome of Ae. spelioides was most similar to the B and G genomes of polyploid wheats on the basis of both C-banding and ISH patterns, but was different from other species of section Sitopsis. These species had different C-banding patterns but they were similar to each other and to Ae. mutica in the distribution of pSc119 hybridization sites. Two types of labeling were detected in Ae. squarrosa with the pAs1 probe. The first resembled that of the D-genome chromosomes of bread wheat, Triticum aestivum L. em. Thell., while the second was similar to the D genome of some of the polyploid Aegilops species. Relationships among diploid Aegilops species and the possible mechanisms of genome differentiation are discussed. Key words : wheat, Triticum, Aegilops, in situ hybridization, C-banding, evolution.


The role of penile color-duplex ultrasound for the evaluation of erectile dysfunction.
In the era of orally active agents, dynamic penile color-duplex ultrasound (D-PCDU) is not considered a necessary first screening for all patients with erectile dysfunction (ED). Various parameters, such as peak systolic flow velocity, end diastolic velocity, resistance index, acceleration time, and degree of arterial dilatation, have been suggested for the diagnosis of vascular ED by D-PCDU. To highlight the clinical utility and evidence-based interpretation of D-PCDU criteria. Extensive, unsystematic PubMed literature search reviewing relevant data on D-PCDU in the evaluation of male ED. The advantage of ultrasound is the minimally invasive nature of the procedure and the ability to screen patients to identify a normal arterial response of cavernous arteries. Men with sexual dysfunctions above 55 years of age and comorbidities are more likely to have multi-organ vascular dysfunction and may necessitate further testing because erectile failure may be the first presenting symptom requiring investigation and treatment even in the absence of cardiovascular risk factors. All patients affected with Peyronie's disease and younger men with persistent ED, a history of pelvic traumas, or fractures of the penile shaft should be offered ultrasonographic penile blood flow studies before referral to surgery or more invasive vascular investigations. In the near future, D-PCDU may be used in preference to patients presenting with or without vascular risk factors, particularly those not responding to first-line orally active drugs and seeking an explanation as to why these agents failed.


Systemic movement of an RNA plant virus determined by a point substitution in a 5' leader sequence.
The ability of viruses to move through infected plants is an important determinant of host range and pathogenicity. We have investigated the genetic basis for the inability of the Type strain of barley stripe mosaic hordeivirus to undergo long-range systemic movement in the tobacco Nicotiana benthamiana. We show that, in this model system, a short open reading frame in the 5' leader of the smallest viral genomic RNA prevents long-range vascular movement. As predicted by the ribosome scanning model, the leader open reading frame decreases the efficiency with which the 5'-proximal gene is translated in vitro. Thus, systemic pathogenicity in this system may be determined by the efficiency of translation of a viral gene in vivo and is not determined by the primary sequence of the encoded protein.


Impact of the SARS-CoV-2 pandemic on lung cancer survival in England: an analysis of the rapid cancer registration dataset.
Early changes in lung cancer care can affect survival. Given the decrease in diagnosis during lockdowns, we calculated their impact on survival using National Lung Cancer Audit data. Percentage survival and HRs for death were compared between 2019 and lockdown periods of 2020. Decreased survival was observed from the first national lockdown onwards and within 90 days of diagnosis. HRs were highest for people diagnosed at the end of 2020 at 1.26 (95% CI 1.20 to 1.32) for death within 90 days and 1.51 (95% CI 1.42 to 1.60) for death between 91 and 270 days. Further work is needed on measures to mitigate this impact.


Non-redundant roles for Profilin2 and Profilin1 during vertebrate gastrulation.
Gastrulation is a critical morphogenetic event during vertebrate embryogenesis, and it is comprised of directional cell movement resulting from the polarization and reorganization of the actin cytoskeleton. The non-canonical Wnt signaling pathway has emerged as a key regulator of gastrulation. However, the molecular mechanisms by which the Wnt pathway mediates changes to the cellular actin cytoskeleton remains poorly defined. We had previously identified the Formin protein Daam1 and an effector molecule XProfilin1 as links for Wnt-mediated cytoskeletal changes during gastrulation. We report here the identification of XProfilin2 as a non-redundant and distinct effector of Daam1 for gastrulation. XProfilin2 interacts with FH1 domain of Daam1 and temporally interacts with Daam1 during gastrulation. In the Xenopus embryo, XProfilin2 is temporally expressed throughout embryogenesis and it is spatially expressed in cells undergoing morphogenetic movement during gastrulation. While we have previously shown XProfilin1 regulates blastopore closure, overexpression or depletion of XProfilin2 specifically affects convergent extension movement independent of mesodermal specification. Specifically, we show that XProfilin2 modulates cell polarization and axial alignment of mesodermal cells undergoing gastrulation independent of XProfilin1. Together, our studies demonstrate that XProfilin2 and XProfilin1 are non-redundant effectors for Daam1 for non-canonical Wnt signaling and that they regulate distinct functions during vertebrate gastrulation.


Vitamin D, multiple sclerosis and inflammatory bowel disease.
It has now been more than 20years since the vitamin D receptor was identified in cells of the immune system. The immune system has now been established as an important target of vitamin D. Vitamin D receptor knockout and vitamin D deficient mice have a surplus of effector T cells that have been implicated in the pathology of multiple sclerosis (MS) and inflammatory bowel disease (IBD). The active form of vitamin D directly and indirectly suppresses the function of these pathogenic T cells while inducing several regulatory T cells that suppress MS and IBD development. There is reason to believe that vitamin D could be an environmental factor that may play a role in the development of these immune mediated diseases in the clinic but at present there has not been a causal relationship established. Nonetheless, current evidence suggests that improving vitamin D status and/or using vitamin D receptor agonists may be useful in MS and IBD.


Outcomes of pregabalin in lumbar-disease patients with depression.
The anti-seizure drug pregabalin is currently used for peripheral neuropathic pain, including degenerative lumbar disease with a neuropathic component. Although there are many reports associated with pregabalin, treatment outcome in low back pain (LBP) patients with depression remains uncertain. This study investigated the outcomes of pregabalin in LBP patients with depression. We assessed 64 patients (29 men and 35 women) using a Visual Analogue Scale, a Self-Rating Depression Scale (SDS) and the Oswestry Disability Index (ODI). Mean age was 63.3 years (range 20-81), and mean duration of disease was 69.8 months (range 3-576). The patients were divided into two groups according to SDS: normal (n = 37) and depressed group (n = 27). Pregabalin significantly reduced both SDS and ODI in the depressed group (p < 0.05). Effect size was larger for both SDS and ODI in the depressed group than in the normal group. Pain was significantly relieved, even in the depressed group (p < 0.01). Pain reduction was achieved by the direct effect of pregabalin, as well as indirect effects attributed to change in depressive symptoms. Although both somnolence and dizziness were detected, the use of hypnotic agents was decreased in half of cases. Somnolence did not influence the analgesic effects of the drug or psychotic state. This investigation indicated that pregabalin is safe and effective for reducing both LBP and mood disturbance in patients with depression.


Validation of a neural network approach for STR typing to replace human reading.
A typical forensic laboratory process for interpreting STR capillary electrophoresis profile data is for two people to independently 'read' the profiles, compare results, and resolve any differences. Recently, work has been conducted to develop a machine learning tool called an artificial neural network (ANN) to carry out the same function as a human profile reader, by classifying areas of fluorescence in the capillary electrophoresis profile raw signal data. The ANN approach has been embedded in commercial software FaSTR™ DNA to read GlobalFiler™ DNA profiles. The ANN feature of FaSTR™ DNA was investigated during validation at Forensic Science South Australia (FSSA) to determine whether one of the human profile readers could be replaced by an ANN reader. FaSTR™ DNA accuracy in detecting allele peaks in reference profiles was 99.7% and was deemed high enough that a one-reader workflow could be implemented into the reference reading workflow at FSSA.


[IgG and IgA levels determination in semen liquid: interest for the biological investigation of infertility].
We studied the relationship between spermogram parameters and biochemical elements in seminal plasma. In this way we searched an association between immunoglobulin levels in seminal plasma and spermogram parameters. The spermogram is usually performed in our laboratory for investigation of infertility. Using immunonephelometry, we measured immunoglobulin (Ig) G and A levels in the seminal plasma of 17 patients. Averages levels of IgG and IgA were 83.3 and 33.6 mg/L, respectively. IgG and IgA levels appeared to be significantly different (p = 0.001), and were significantly associated. A correlation between IgG values and coiled tails percentage was noted in patients having IgA below 20 mg/L (p < 0,001). In conclusion, the measurement of IgG and IgA levels in seminal plasma might constitute a valuable diagnosis tool for the evaluation of infertility.


Cognitive status correlates with neuropathologic stage in Parkinson disease.
To study the association of cognitive status with the stages of a published neuropathologic staging procedure for sporadic Parkinson disease (PD) in a cohort of 88 patients with PD from a single neurologic unit. None had received the clinical diagnosis of dementia with Lewy bodies (DLB). The authors assessed Lewy neurites/bodies (LNs/LBs) immunoreactive for alpha-synuclein semiquantitatively in sections from 18 brain regions. In silver-stained sections and sections immunostained for tau and beta-amyloid protein, the authors semiquantitatively evaluated comorbidities potentially contributing to cognitive decline, e.g., Alzheimer disease (AD), argyrophilic grain disease (AGD), and cerebral vascular disease. The authors analyzed four Mini-Mental State Examination (MMSE) subgroups ranging from marginally impaired cognition to severe dementia using nonparametric tests. It was possible to assign all patients to one of the PD stages. MMSE scores correlated with neuropathologic stages (p < 0.005) and this association showed a linear trend (p < 0.025). Median MMSE test scores for women were lower than those for men. Cognitively impaired individuals displayed higher stages of AD-related neurofibrillary pathology (p < 0.05) and beta-amyloid deposition (p < 0.05) than cognitively unimpaired persons. MMSE scores did not correlate significantly with AGD, disease duration, age at disease onset, or age at death. Hoehn and Yahr scores, however, correlated with PD stages (p < 0.0005) and MMSE scores (p < 0.0005). The decrease in median Mini-Mental State Examination scores between PD stages 3 to 6 indicates that the risk of developing dementia increases with disease progression. In some individuals, however, cognitive decline can develop in the presence of mild Parkinson disease-related cortical pathology and, conversely, widespread cortical lesions do not necessarily lead to cognitive decline.


Limitations of intrathecal baclofen for spastic hemiparesis following stroke.
Intrathecal baclofen (ITB) has become the first choice in the management of deleterious spasticity that does not respond to oral and intramuscular medications following spinal cord injury, traumatic brain injury, and cerebral palsy. The usefulness of ITB in severe spastic hemiparesis following stroke is studied. A total of 8 patients underwent clinical and video assessment following ITB bolus application (n=5) and during continuous infusion via a temporary catheter system (n=3). The mean daily dosage alleviating spasticity on the hemiparetic side-without affecting the nonparetic side-was 119 microg/day (range 50 to 360 microg/day). However, 6 patients experienced functional deterioration as ITB weakened their paretic side such that the antigravity pattern they used for ambulation was suppressed. In 2 patients, spasticity-associated pain and spasms subsided and they underwent implantation of a long-term drug delivery system. ITB may reduce spasticity in a dose-dependent manner irrespective of its origin. Although not primarily antinociceptive in humans, ITB may alleviate pain if arising from increased muscle tone. A functional benefit may result if ITB can uncover "subclinical" motor control that had been suppressed by spasticity. However, when a patient uses antigravity patterns for ambulation in the absence of more complex motor control, ITB may cause the loss of residual walking ability, which becomes a major limitation for patients with hemiplegic stroke.


Identification and partial characterization of multiple forms of biologically active EGF in rat milk.
Milk from lactating Sprague-Dawley rats was assayed for epidermal growth factor (EGF)-like activities. A homologous radioimmunoassay (RIA) indicated the presence of immunoreactive material that competed in a nonparallel fashion with submandibular gland rat EGF (sm-r-EGF). The activity in the milk was extracted using antibodies to sm-r-EGF covalently linked to acrylamide beads. This activity was characterized by RIA, nondenaturing polyacrylamide gel electrophoresis, enzymatic digestion, radioreceptor assay, and ability to stimulate incorporation of [3H]thymidine into cultured fibroblasts. The presence of three distinct immunoreactive forms of EGF in rat milk were detected that competed with 125I-labeled r-EGF for binding to the EGF receptor and stimulated DNA synthesis in growth-arrested fibroblasts. Two of the forms are converted to the sm-r-EGF species by tryptic digestion as determined by RIA and migration rates in a nondenaturing polyacrylamide gel. The biological activities are stable to heating in 0.1 M acetic acid and also are stable at pH 9.0. Levels of r-EGF equivalents in milk were low at time of birth (6.3 +/- 1.7 ng/ml) and rose to 35.4 +/- 14.6 by days 4 to 6.


Health-related quality of life in transplant ineligible newly diagnosed multiple myeloma patients treated with either thalidomide or lenalidomide-based regimen until progression: a prospective, open-label, multicenter, randomized, phase 3 study.
Data on the impact of long term treatment with immunomodulatory drugs (IMiD) on health-related quality of life (HRQoL) is limited. The HOVON-87/NMSG18 study was a randomized, phase 3 study in newly diagnosed transplant ineligible patients with multiple myeloma, comparing melphalan-prednisolone in combination with thalidomide or lenalidomide, followed by maintenance therapy until progression (MPT-T or MPR-R). The EORTC QLQ-C30 and MY20 questionnaires were completed at baseline, after three and nine induction cycles and six and 12 months of maintenance therapy. Linear mixed models and minimal important differences were used for evaluation. 596 patients participated in HRQoL reporting. Patients reported clinically relevant improvement in global quality of life (QoL), future perspective and role and emotional functioning, and less fatigue and pain in both arms. The latter being of large effect size. In general, improvement occurred after 6-12 months of maintenance only and was independent of the World Health Organisation performance at baseline. Patients treated with MPR-R reported clinically relevant worsening of diarrhea, and patients treated with MPT-T reported a higher incidence of neuropathy. Patients who remained on lenalidomide maintenance therapy for at least three months reported clinically meaningful improvement in global QoL and role functioning at six months, remaining stable thereafter. There were no clinically meaningful deteriorations, but patients on thalidomide reported clinically relevant worsening in neuropathy. In general, HRQoL improves both during induction and maintenance therapy with immunomodulatory drugs. The side effect profile of treatment did not negatively affect global QoL, but it was, however, clinically relevant for the patients. (Clinicaltrials.gov identifier: NTR1630).


T-kininogenase activity of the rat submandibular gland is predominantly due to the kallikrein-like serine protease antigen gamma.
T-kininogen, the major kininogen in rat plasma, releases Ile-Ser-bradykinin (T-kinin) when incubated with trypsin, but is not a substrate for tissue kallikrein. Enzymes able to release T-kinins from T-kininogen have been found in the rat submandibular gland, but precise identification of these enzymes and their possible relationship to kallikrein-like enzymes has not been established. We studied T-kininogenase activity in fractionated submandibular gland homogenate. The main T-kininogen catalytic enzyme was purified and characterized, and found to be identical to antigen gamma, a kallikrein-like enzyme which we have previously characterized. Of other identified kallikrein-like enzymes only tonin showed weak T-kininogenase activity, which was about 0.25% of that of antigen gamma. No other T-kininogen catalytic enzymes were observed. Antigen gamma released a kinin which was identified as T-kinin by reverse-phase h.p.l.c. The T-kininogenase activity of antigen gamma had a Km of 29 +/- 4 microM and a kcat/Km of 140 M-1.s-1, and was comparable with its high and low molecular mass-kininogenase activity (7.4 and 10 micrograms of kinin/h per mg respectively). In contrast, tissue kallikrein released 0.2 and 42,200 micrograms of kinin/h per mg respectively. Thus antigen gamma is a weak kininogenase. The isoelectric point of antigen gamma, but not its molecular mass, differed from that of other kallikrein-like enzymes. Isoelectrofocusing in flat-bed gels combined with immunostaining was therefore a convenient method for identification. The kallikrein-like nature of antigen gamma was demonstrated by its immunological similarity to tissue kallikrein and tonin and by 91% and 87% amino acid sequence similarity with tonin and kallikrein respectively (67 amino acids sequenced). Complete identity was also not observed with other sequenced kallikrein genes, mRNAs or proteins.


Blunt thoracic aortic injury: delayed or early repair?
To evaluate the safety and benefit of delayed repair of blunt thoracic aortic injury (BTAI) in trauma patients with multiple injuries and to assess the financial impact of delayed repair. A retrospective review of charts was performed on 55 patients with the diagnosis of BTAI from January 1, 1992, through December 31, 1997, at our Level I trauma center. Early repair was defined as operative repair of BTAI within 12 hours of admission. Seven patients were excluded from analysis due to death before BTAI diagnosis (two deaths were from rupture in the emergency department and five were from massive blunt trauma without rupture). The groups were compared by using a McNemar chi2 test, for which p less than or equal to 0.05 is significant. There were 30 patients in the early repair (ER) group repaired at 5.3+/-2.4 hours, and 18 patients in the delayed repair (DR) group repaired at 8.5 days (range, 17 hours-67 days). There were no significant differences between the ER and DR groups in age (37+/-18 years vs. 41+/-19 years), Injury Severity Score (39+/-15 vs. 45+/-14), intensive care unit days (12+/-14 days vs. 18+/-11 days), hospital length of stay (21+/-19 days vs. 28+/-14 days), or mortality rates (7% vs. 6%). There was a trend toward longer lengths of stay in the DR group. Most DR patients required beta-blocker therapy and/or other antihypertensives for systolic BP more than 120 mm Hg during admission. There were no deaths from aortic rupture in either group. By using financial data that was available from July of 1994 onward, we performed a subset analysis of the direct costs associated with BTAI. Total direct and variable direct costs for patients undergoing delayed repair were over two times the costs for early repair patients (p < 0.05). The management of trauma patients with multiple injuries requires prioritization of injuries so that the outcomes from these injuries can be optimized. Although delayed aortic repair was safely practiced in this series, there was not an obvious outcome benefit to delayed repair. The patients undergoing late repair required increased attention to hemodynamics, and there was a trend toward increased length of stay. In addition, analysis of the costs associated with delayed repair demonstrated a twofold increase in the direct costs for delayed repair compared with early repair.


Toxicity testing in the 21st century beyond environmental chemicals.
After the publication of the report titled Toxicity Testing in the 21st Century - A Vision and a Strategy, many initiatives started to foster a major paradigm shift for toxicity testing - from apical endpoints in animal-based tests to mechanistic endpoints through delineation of pathways of toxicity (PoT) in human cell based systems. The US EPA has funded an important project to develop new high throughput technologies based on human cell based in vitro technologies. These methods are currently being incorporated into the chemical risk assessment process. In the pharmaceutical industry, the efficacy and toxicity of new drugs are evaluated during preclinical investigations that include drug metabolism, pharmacokinetics, pharmacodynamics and safety toxicology studies. The results of these studies are analyzed and extrapolated to predict efficacy and potential adverse effects in humans. However, due to the high failure rate of drugs during the clinical phases, a new approach for a more predictive assessment of drugs both in terms of efficacy and adverse effects is getting urgent. The food industry faces the challenge of assessing novel foods and food ingredients for the general population, while using animal safety testing for extrapolation purposes is often of limited relevance. The question is whether the latest paradigm shift proposed by the Tox21c report for chemicals may provide a useful tool to improve the risk assessment approach also for drugs and food ingredients.


Anger and shame as determinants of perceived competence.
Emotions are complex signals conveying a multitude of "messages" concomitantly. This idea is examined within the context of competence inferences drawn from the emotional expressions of another individual. In two studies, participants assuming the role of patients took part in a simulated medical consultation. They encountered a physician who had either a high or a standard professional status, and who responded with anger, shame, or emotional neutrality when asked to clarify the advice he dispensed. While a display of anger did not affect perceived competence, shame made the physician appear less competent. Three types of signals conveyed by the emotions were responsible for these effects: the physician's decisiveness and control over the situation, and the extent to which he felt professionally devalued by the patient's request, mediated the effects of the emotions on perceived competence. A priori information about the physician's professional status had little effect on the perception of competence. The research exemplifies the richness of information contained in emotions, and the complex way in which it allows observers to construe an impression of the expresser.


Adipose-derived regenerative cell therapy inhibits the progression of monocrotaline-induced pulmonary hypertension in rats.
Functional and structural changes in pulmonary vasculature characterize pulmonary arterial hypertension (PAH) and the prognosis of advanced PAH remains poor despite progress in pharmacotherapy. Adipose-derived regenerative cells (ADRCs) promote cell regeneration at pathological sites and comprise a novel therapy for ailments of various organs. We investigated the effects of ADRC therapy in rat models of monocrotaline (MCT)-induced pulmonary hypertension (PH) and the underlying mechanisms. Rats were assigned to Control and MCT groups without and with (M/A) intravenous transfusion of seven million ADRCs on day 7. We echocardiographically evaluated pulmonary hypertension as pulmonary artery flow acceleration time (PAAT) and deceleration (PADc). Right ventricular (RV) systolic pressure was measured by catheterization on day 28 and then pathological changes in pulmonary vessels were assessed. We analyzed PAH-associated gene expression on day 14 using real-time RT-PCR. Echocardiography and RV catheterization showed that ADRC therapy inhibited PH development (assessed as PAAT, PADc, and RV systolic pressure) at day 28 (MCT vs. M/A, P<0.05). Pulmonary vascular remodeling was also inhibited (vessel wall thickness: MCT vs. M/A, P<0.01). Messenger RNA levels of endothelin (ET) A and B receptors, ET-1 and transforming growth factor (TGF)-β increased in the lungs by MCT were suppressed by ADRCs (MCT vs. M/A, P<0.05). The development of PH was inhibited by ADRCs through suppressing changes in the expression of genes associated with ET and TGF-β systems. We believe that ADRC therapy could serve as a novel strategy for treating PH.


Development, psychometric evaluation, and initial feasibility assessment of a symptom tracker for use by patients with heart failure (HFaST).
This study aimed to develop and provide a psychometric and feasibility pilot evaluation of the Heart Failure (HF) Symptom Tracker (HFaST), a new patient-reported tool designed to facilitate communication between patients and health care providers (HCPs) in routine clinical care. The HFaST enables patients to identify worsening HF symptoms, with a long-term goal of preventing hospitalizations or emergency room visits. The HFaST was developed drawing on evidence from the literature, qualitatively with cognitive interviews (12 patient/caregiver and 8 HCPs), and evaluated quantitatively (psychometric, feasibility assessment). The HFaST was administered for 7 consecutive days to 100 individuals diagnosed with HF during a multisite, non-interventional US pilot study. Health care providers then completed a survey assessing the feasibility and importance of the HFaST in clinical practice. Qualitative development included a literature review and cognitive interviews with patients, caregivers, and HCPs. The psychometric properties of the HFaST were evaluated using classical test theory methods. Descriptive statistics provided insight into HCPs' perceptions of the feasibility of using the HFaST in clinical practice. A preliminary set of 40 items was developed for the symptom tracker and iteratively reduced to 10 items based on the qualitative phase. Test-retest reliability (weighted kappa 0.71-0.97), discriminating validity, and construct validity of the HFaST were acceptable. HCPs rated the HFaST as a good (70%) or excellent (30%) means of tracking HF symptoms. Six HFaST items were ultimately retained, covering concepts of fatigue, shortness of breath (3 items), swelling, and rapid weight gain. The 6-item HFaST is an easy-to-use tool designed to raise patients' awareness of HF symptoms and facilitate communication with HCPs. Future research should evaluate HFaST implementation in clinical practice and effectiveness as an intervention to potentially prevent hospitalizations and emergency room visits.


Germline Amino Acid Diversity in B Cell Receptors is a Good Predictor of Somatic Selection Pressures.
The diversity of the immune repertoire is important for the adaptive immune system's ability to detect pathogens. Much of this diversity is generated in two steps, first through the recombination of germline gene segments and second through hypermutation during an immune response. While both steps are to some extent based on the germline level repertoire of genes, the final structure and selection of specific receptors is at the somatic level. How germline diversity and selection relate to somatic diversity and selection has not been clear. To investigate how germline diversity relates to somatic diversity and selection, we considered the published repertoire of Ig heavy chain V genes taken from the blood of 12 individuals, post-vaccination against influenza, sequenced by 454 high-throughput sequencing. We here show that when we consider individual amino acid positions in the heavy chain V gene sequence, there exists a strong correlation between the diversity of the germline repertoire at a position and the number of B cell clones that change amino acids at that position. At the same time, we find that the diversity of amino acids used in the mutated positions is greater than in the germline, albeit still correlated to germline diversity. From these findings, we propose that while germline diversity and germline amino acid usage at a given position do not fully specify the amino acid mutant needed to promote survival of specific clones, germline diversity at a given position is a good indicator for the potential to survive after somatic mutation at that position. We would therefore suggest that germline diversity at each specific position is the better a priori model for the effects of somatic mutation and selection, than simply the division into complementarity determining and framework regions.


Secretome-based identification of TFPI2, a novel serum biomarker for detection of ovarian clear cell adenocarcinoma.
Of all of the epithelial ovarian cancers (EOC), clear cell adenocarcinoma (CCA) has the worst clinical prognosis. Furthermore, the conventional EOC biomarker CA125 is more often negative in CCA than in other subtypes of EOC. This study sought to discover a new diagnostic biomarker that would allow more reliable detection of CCA. Using mass spectrometry, we compared proteins in conditioned media from cell lines derived from CCA and other types of EOC. We identified 30 extracellular or released proteins specifically present in CCA-derived cell lines. Bioinformatics analyses identified a serine protease inhibitor, tissue factor pathway inhibitor 2 (TFPI2), as a potential biomarker for CCA. Real time RT-PCR and Western blot analyses revealed that TFPI2 was exclusively expressed in CCA-derived cell lines and tissues. For clinical validation, we measured levels of TFPI2 and CA125 in a set of sera from 30 healthy women, 30 patients with endometriosis, and 50 patients with CCA, using an automated enzyme-linked immunosorbent assay systems. Serum levels of TFPI2 were significantly elevated in CCA patients, even those with normal CA125 levels. In terms of area under the receiver operating characteristic curve (AUC), TFPI2 was superior to CA125 in discriminating CCA patients from healthy women (AUC 0.97 for TFPI2 versus AUC 0.80 for CA125), or from patients with endometriosis (AUC 0.93 for TFPI2 versus 0.80 for CA125). This is the first evidence for TFPI2 as a serum biomarker of CCA. We propose that this biomarker may be useful for detection of CCA and for monitoring the transformation from endometriosis into CCA.


A fast and fatal course of bronchiectasis: an unusual rare expression of chronic graft versus host disease. A case report.
Introduction. We report a case of a patient with acute myeloid leukaemia whose treatment with bone marrow transplantation (BMT) was followed by chronic graft versus host disease (GVHD) with lung involvement and bronchiectasis. This report illustrates an unusual course of a fast progression of the bronchiectasis due to BMT. Case description. A 33-year-old female was diagnosed with acute myeloid leukaemia. An allogeneic BMT was performed. One month after the transplantation, acute GVHD with skin involvement occurred. Treatment with prednisolone and mycophenolate mofetil (MMF) has been started. Nine months later, the patient was examined by a pulmonologist due to progressive dyspnoea. A pulmonary computed tomography (CT) scan showed normal parenchyma of the lungs and no changes to the bronchi. A CT scan performed 7 months later revealed bronchiectasis for the first time. No clinical response was associated with the treatment and the patient's respiratory status progressively deteriorated. During the final hospitalization, a CT scan performed 1 year later revealed huge cystic bronchiectasis in both lungs. Despite the prophylaxis and treatment of GVHD and aggressive antimicrobial therapy, the patient died one year after the diagnosis of bronchiectasis. Conclusions. This case demonstrates that a fast and fatal course of bronchiectasis, that occurs after BMT, should always be considered as a possible manifestation of chronic graft versus host disease (cGVHD) following allogeneic BMT.


Comparison of continuously acquired resting state and extracted analogues from active tasks.
Functional connectivity analysis of brain networks has become an important tool for investigation of human brain function. Although functional connectivity computations are usually based on resting-state data, the application to task-specific fMRI has received growing attention. Three major methods for extraction of resting-state data from task-related signal have been proposed (1) usage of unmanipulated task data for functional connectivity; (2) regression against task effects, subsequently using the residuals; and (3) concatenation of baseline blocks located in-between task blocks. Despite widespread application in current research, consensus on which method best resembles resting-state seems to be missing. We, therefore, evaluated these techniques in a sample of 26 healthy controls measured at 7 Tesla. In addition to continuous resting-state, two different task paradigms were assessed (emotion discrimination and right finger-tapping) and five well-described networks were analyzed (default mode, thalamus, cuneus, sensorimotor, and auditory). Investigating the similarity to continuous resting-state (Dice, Intraclass correlation coefficient (ICC), R(2) ) showed that regression against task effects yields functional connectivity networks most alike to resting-state. However, all methods exhibited significant differences when compared to continuous resting-state and similarity metrics were lower than test-retest of two resting-state scans. Omitting global signal regression did not change these findings. Visually, the networks are highly similar, but through further investigation marked differences can be found. Therefore, our data does not support referring to resting-state when extracting signals from task designs, although functional connectivity computed from task-specific data may indeed yield interesting information.


A predictive model of hospitalisation and death from chronic obstructive pulmonary disease.
A recent study showed that doctors are excessively pessimistic about the prognosis in patients with COPD and suggested that a simple tool to predict outcome is needed. In a prospective observational study, 3343 patients with an FEV1<80% of the predicted value and FEV1/FVC<70% were selected from a clinical network of patients screened for COPD in Tayside, Scotland. Data were collected during annual visits on demography, spirometry, smoking history, medical research council (MRC) dyspnoea scale, body mass index (BMI) and other variables. The main outcome measures were hospitalisations and death secondary to COPD. A proportional hazard model was used to identify significant risk factors. Increasing age, low BMI, worsening MRC dyspnoea score, decreased FEV1, and prior respiratory or cardiovascular admission hospitalisation were predictors of poor outcome. Influenza vaccination was protective. We have developed a model that estimates the risk of respiratory hospitalisation and death in patients with COPD.


A Potential Probiotic for Diarrhea: Clostridium tyrobutyricum Protects Against LPS-Induced Epithelial Dysfunction via IL-22 Produced By Th17 Cells in the Ileum.
Probiotics are clinically used for diarrhea and inflammatory bowel diseases in both humans and animals. Previous studies have shown that Clostridium tyrobutyricum (Ct) protects against intestinal dysfunction, while its regulatory function in the gut needs further investigation and the related mechanisms are still not fully elucidated. This study aims to further verify the protective function of Ct and reveal its underlying mechanisms in alleviating diarrhea and intestinal inflammation. Ct inhibited LPS-induced diarrhea and intestinal inflammation in the ileum. IL-22 was identified and the protective role of Ct in the ileum presented an IL-22-dependent manner according to the transcriptomic analysis and in vivo interference mice experiments. The flow cytometric analysis of immune cells in the ileum showed that Ct enhanced the proportions of Th17 cells in response to LPS. The results of in situ hybridization further verified that Ct triggered Th17 cells to produce IL-22, which combined with IL-22RA1 expressed in the epithelial cells. Moreover, Ct was unable to enhance the levels of short-chain fatty acids (SCFAs) in the ileum, suggesting that the protective role of Ct in the ileum was independent of SCFAs. This study uncovered the role of Ct in alleviating diarrhea and inflammation with the mechanism of stimulating Th17 cells in the lamina propria to produce IL-22, highlighting its potential application as a probiotic for diarrhea and inflammation in the ileum.


Pleiotropic effect of the rec A gene of Escherichia coli: uncoupling of cell division from deoxyribonucleic acid replication.
A defective recA gene, which is involved in recombination, is shown in this article to permit limited cell division, when deoxyribonucleic acid (DNA) synthesis is blocked. Thymidine starvation or nalidixic acid blocked DNA synthesis, and stopped cell division of a rec(+)thy(-) strain of Escherichia coli. However, with the same treatments, a recAthy(-) strain could continue to divide for at least 5 hr, and cell numbers increased 2.5- to 4-fold. After several hours of thymidine starvation, the culture contained very long cells (snakes) and small (normal-sized) cells. The short cells contained very little, if any, DNA. Cells of all ages divided in the absence of thymidine. Specific differences in membrane proteins were observed between thymidine-starved rec(+) and recA cells, as expected from previous experiments in which these proteins were associated with cell division and DNA synthesis. It is proposed that septum formation is controlled negatively by the recA(+) gene.


Lung Ultrasound Score in Evaluating the Severity of Coronavirus Disease 2019 (COVID-19) Pneumonia.
The purpose of this study is to observe the potential of lung ultrasound in evaluating the severity of coronavirus disease 2019 (COVID-19) pneumonia. Lung ultrasound was performed in ten zones of the patients' chest walls. The features of the ultrasound images were observed, and a lung ultrasound score (LUS) was recorded. The ultrasound features and scores were compared between the refractory group (PaO2/FiO2 ≤ 100 mm Hg or on extracorporeal membrane oxygenation) and the non-refractory group. The prediction value of the LUS was studied by receiver operating characteristic (ROC) curve analysis. In total, 7 patients were enrolled in the refractory group and 28 in the non-refractory group. B-line patterns and shred signs were the most common signs in all patients. Patients in the refractory group had significantly more ground-glass signs (median 6 [interquartile range {IQR}, 2.5-6.5] vs. median 0 [IQR, 0-3]), consolidation signs (median 1 [IQR, 1-1.5] vs. median 0 [IQR, 0-3]) and pleural effusions (median 5 [IQR, 1.5-6] vs. median 0 [IQR, 0-0.25]). The LUS was significantly higher in the refractory group (33.00 [IQR 27.50-34.00] vs. 25.50 [IQR 22.75-30.00]). The ROC of the LUS showed a cutoff score of 32 with a specificity of 0.893 and a sensitivity of 0.571 in diagnosing refractory respiratory failure among patients. In COVID-19 patients, lung ultrasound is a promising diagnostic tool in diagnosing patients with refractory pneumonia.


A network-based, integrative approach to identify genes with aberrant co-methylation in colorectal cancer.
Epigenetic changes, including aberrations in DNA methylation, are a common hallmark of many cancers. The identification and interpretation of epigenetic changes associated with cancers may benefit from integration with protein interactomes. Based on the assumption that genes implicated in a specific tumor phenotype will show high aberrant co-methylation patterns with their interacting partners, we propose an integrated approach to uncover cancer-associated genes by integrating a DNA methylome with an interactome. Aberrant co-methylated interactions were first identified in the specific cancer, and genes were then prioritized based on their enrichment in aberrant co-methylation. By applying this to a large-scale colorectal cancer (CRC) dataset, the proposed method increases the power to capture known genes. More importantly, genes possessing high aberrant co-methylation patterns, located at the topological center of the original protein-protein interaction network (PPIN), affect several cancer-associated pathways and form hotspots that are frequently hijacked in cancer. Additionally, the top-ranked candidate genes may also be useful as an indicator of CRC diagnosis and prognosis. Five fold cross-validation of the top-ranked genes in diagnosis reveals that it can achieve an area under the receiver operating characteristic (ROC) curve ranging from 82.2% to 98.4% in three independent datasets. Five of these genes form a core repressive module. CCNA1 and ESR1 in particular are evidently silenced by promoter hypermethylation in CRC cell lines and tissues, whose re-expression markedly suppresses tumor cell survival and clonogenicity. These results show that the network-centric method could identify novel disease biomarkers and model how oncogenic lesions mediate epigenetic changes, providing important insights into tumorigenesis.


Non-IgE mediated food allergy.
Adverse reactions to dietary proteins (DPs) can impose a significant impact on one's daily life and can even affect the 'life style' of an entire family. Adverse reactions to DPs may or may not be immune-mediated. The immune-mediated adverse reaction to food is defined as food allergy (FA) which is roughly divided into IgE mediated or non-IgE mediated FA (NFA). As opposed to IgE mediated FA, NFA primarily affects the GI mucosa. In addition, there is far less of an understanding of NFA than IgE-mediated FA and its clinical relevance is likely under-estimated in most cases. This is partly due to delayed onset of symptoms and subsequent difficulty in making the clinical association between offending food and clinical symptoms. The lack of easily accessible diagnostic measures also contributes to the problem. The gut mucosal barrier is thought to have developed to execute an immensely difficult task; digestion and absorption of nutrients without provoking immune responses and cohabiting with commensal flora in a mutual beneficial relationship, while maintaining an immune defense against pathogenic microbes. The gut mucosal immune system accomplishes this task partly by establishing tolerance to macronutrients with potent immunogenecity. Immune tolerance to macronutrients (DPs) is maintained in part by active suppressive mechanisms involving antigen (Ag)-specific regulatory T (Treg) cells. This active immune tolerance state appears to be affected by various environmental factors such as change in commensal flora. In the first few years of life, humans gradually develop an intricate balance between tolerance and immune reactivity in the gut mucosa along with a tremendous expansion of gut associated lymphoid tissue (GALT). Not surprisingly, both IgE and non-IgE mediated food allergy (FA) is frequently seen during this period. The most common causative DPs for NFA are those contained in infant formulas (cow's milk and soy proteins). Unlike IgE mediated FA, NFA is rarely life-threatening. However, NFA to DPs can cause significant morbidity in rapidly growing infants and young children. A better understanding of pathogenesis of NFA is crucial for timely management of NFA in this vulnerable population. This review discusses the gut mucosal immune system in the first few years of life including genetic/environmental factors affecting the development of mucosal immune system and pathogenesis of NFA in association with clinical/laboratory findings.


HLA-DR15 haplotype and multiple sclerosis: a HuGE review.
An association between multiple sclerosis (MS) and the human leukocyte antigen (HLA) complex, a dense cluster of genes on the short arm of chromosome 6, was first noted over 30 years ago. In Caucasian populations of Northern European descent, the DR15 haplotype (DRB1*1501-DQA1*0102-DQB1*0602) has been hypothesized to be the primary HLA genetic susceptibility factor for MS. However, studies of other populations have produced varying results. Thus, the authors reviewed the literature for articles on the association between the DR15 haplotype and MS. They identified 72 papers meeting the inclusion criteria: human genetic studies written in English that were published between 1993 and 2004 and that reported allele frequencies for HLA-DRB1*1501, HLA-DQA1*0102, or HLA-DQB1*0602 or the frequency of the DRB1*1501-DQA1*0102-DQB1*0602 haplotype. Most of the studies identified used a case-control design (n = 60), while the remainder used a family-based design (n = 22). In most of these papers, investigators reported a higher frequency of the DR15 haplotype and/or its component alleles among MS cases than among controls. However, the authors' confidence in these results is tempered by factors related to study design that may have biased the outcomes.


Interleukin 1 beta and interleukin 6 potentiate retinoic acid-mediated repression of human immunodeficiency virus type 1 replication in macrophages.
All-trans-retinoic acid (RA) has been shown either to activate or repress human immunodeficiency virus type 1 (HIV-1) replication in primary monocyte-derived-macrophages (MDMs). We systematically investigated the contribution that cell donor and virus differences make to this variability. We found that the effect of RA was cell donor dependent. In addition, the ability of RA to repress HIV-1 replication varied between different virus stocks. In no case did RA affect either virus entry or integration but instead affected the accumulation of viral mRNAs in infected cells. Despite the complex variability in RA responsiveness in untreated cells, we found that RA consistently repressed virus replication when the MDMs were treated with concentrations of interleukin 1 beta (IL-1 beta) and IL-6 that are expected at local sites of infection, where HIV-1-infected macrophages reside in vivo.