Decrease in Numbers of Naive and Resting B Cells in HIV-Infected Kenyan Adults Leads to a Proportional Increase in Total and Plasmodium falciparum-Specific Atypical Memory B Cells.
Human immunodeficiency virus type 1 (HIV-1) infection is associated with B cell activation and exhaustion, and hypergammaglobulinemia. How these changes influence B cell responses to coinfections such as malaria is poorly understood. To address this, we compared B cell phenotypes and Abs specific for the Plasmodium falciparum vaccine candidate apical membrane Ag-1 (AMA1) in HIV-infected and uninfected adults living in Kenya. Surprisingly, HIV-1 infection was not associated with a difference in serum AMA1-specific Ab levels. HIV-infected individuals had a higher proportion of total atypical and total activated memory B cells (MBCs). Using an AMA1 tetramer to detect AMA1-specific B cells, HIV-infected individuals were also shown to have a higher proportion of AMA1-specific atypical MBCs. However, this proportional increase resulted in large part from a loss in the number of naive and resting MBCs rather than an increase in the number of atypical and activated cells. The loss of resting MBCs and naive B cells was mirrored in a population of cells specific for an Ag to which these individuals were unlikely to have been chronically exposed. Together, the data show that changes in P. falciparum Ag-specific B cell subsets in HIV-infected individuals mirror those in the overall B cell population, and suggest that the increased proportion of atypical MBC phenotypes found in HIV-1-infected individuals results from the loss of naive and resting MBCs.


A NEW RECOMBINANT ADENO-ASSOCIATED VIRUS (AAV)-BASED RANDOM PEPTIDE DISPLAY LIBRARY SYSTEM: INFECTION-DEFECTIVE AAV1.9-3 AS A NOVEL DETARGETED PLATFORM FOR VECTOR EVOLUTION.
Directed evolution through genetic engineering of viral capsids followed by selection has emerged as a powerful means to create novel recombinant adeno-associated virus (rAAV) vectors with desired tropism and enhanced properties. One of the most effective approaches uses rAAV-based random peptide display libraries. Here we report a novel system based on an infection-defective rAAV1.9-3 as a platform for random peptide display, and show that biopanning of the libraries in vitro effectively identifies the peptides that restore and enhance rAAV transduction. rAAV1.9-3 has a genetically engineered AAV1 capsid with amino acids 445-568 being replaced with those of AAV9, and has been identified as a variant exhibiting significantly impaired infectivity and delayed blood clearance when infused into mice. In this study, we generated rAAV1.9-3 variant libraries in which 7- or 12-mer random peptides were expressed at the capsid amino acid position 590. Three rounds of positive selection for primary human dermal fibroblasts successfully identified new rAAV-peptide variants that transduce them more efficiently than the prototype rAAV2. Thus our study demonstrates that an infection-defective rAAV variant serves as a novel detargeted platform for random peptide display libraries. We also describe a brief review of recent progress in rAAV-based random peptide display library approaches.


Inflammation and infiltration: can the radiologist draw a line? MRI versus CT to accurately assess medullary involvement in parosteal osteosarcoma.
Cancer causes inflammation as it progresses through healthy tissue. The differentiation of tumoral growth from the surrounding inflammatory change is paramount in planning surgeries seeking to preserve function. This retrospective study aims at illustrating how a careful use of imaging (computed tomography (CT)/magnetic resonance imaging (MRI)) can help to draw the line between infiltration and inflammation. Out of 72 cases of parosteal osteosarcoma in our institution we selected 22 which had pretreatment imaging, and out of those, 14 that had both MRI and CT. Using Fisher's exact test, we evaluated the performance of each technique on accurately diagnosing medullary tumor infiltration, using histological analysis as a gold standard. All cases (14/14) demonstrated medullary abnormality on MRI, but only 6/14 (42.9%) demonstrated abnormality on CT. The 8/14 cases with MRI abnormality but no CT abnormality (57.1%) showed inflammation with no tumoral cells present on histological analysis. In the cases where the two examinations showed medullary abnormality (6/14) histology demonstrated tumoral infiltration. MRI demonstrated high sensitivity and negative predictive value, but low specificity and low positive predictive value and accuracy (P=1). CT demonstrated high sensitivity, specificity, high positive and negative predictive values and accuracy (P = 0.000333). MRI is highly sensitive for the detection of medullary abnormality but lacks specificity for tumor invasion. Correlation with CT is recommended in all cases of positive MR to add specificity for tumors. The adequate use of the two imaging methods allows to differentiate between inflammatory change and tumoral infiltration in POS, relevant for surgical planning.


Differences in breast cancer incidence and stage distribution between urban and rural female population in Podlaskie Voivodship, Poland in years 2001-2002.
The aim of the study was to evaluate differences in breast cancer incidence and stage of disease between the urban and rural female population in Podlaskie Voivodship in 2001-2002, before the introduction of the Population Screening Programme in 2006. Analysis was based on 696 breast cancer cases diagnosed in years 2001-2002 and registered in the CR in Bialystok (Voivodship Cancer Registry). An average annual number of incidence, as well as crude and standardised incidence rates, were calculated. Age-specific incidence rates for 5-year age groups were also calculated and grouped as follows: < 50, 50-69, > =70 years old. Incidence differences related to place of residence: urban or rural, were presented with the use of u/r (urban/rural) ratio. In order to evaluate the stage of disease, a simplified classification recommended by ENCR (European Network of Cancer Registries) for population registries (localised, regional, metastatic) was applied. The breast cancer incidence rate in the urban population was higher than in rural areas with u/r ratio amounting to 1.4. The highest incidence and largely marked differences between urban and rural areas were among women aged 50-69 years with the u/r ratio amounting to 1.8. Overall, the proportion of stage localised in Podlaskie Voivodship was 33.1 percent and differed between urban and rural areas. The proportion of localised cancer was higher in urban areas, but patients were younger when compared to those living in rural areas. Knowledge of differences in incidence and breast cancer stage in urban and rural women investigated in this research, together with other epidemiological indicators, should be used for monitoring the Population Screening Programmes in these populations.


Liver collagen proportionate area predicts decompensation in patients with recurrent hepatitis C virus cirrhosis after liver transplantation.
Current histological scoring systems do not subclassify cirrhosis. Computer-assisted digital image analysis (DIA) of Sirius Red-stained sections measures fibrosis morphologically producing a fibrosis ratio (collagen proportionate area [CPA]). CPA could have prognostic value within a disease stage, such as cirrhosis. The aim of the present study was to evaluate CPA in patients with recurrent hepatitis C virus (HCV) allograft cirrhosis and assess its relationship with hepatic venous pressure gradient (HVPG). In 121 consecutively-transplanted HCV patients with HVPG, measured contemporaneously with transjugular liver biopsies, 65 had Ishak stage 5 or 6 disease (43 with HVPG measurement). Biopsies were stained with Sirius Red for DIA, and the collagen content was expressed as a CPA. In three cases, a tissue for Sirius Red staining was not obtained, and the patients were excluded. Sixty-two patients were analyzed. The median HVPG was 8 mmHg (interquartile range [IQR]: 5-10). Portal hypertension (HVPG ≥ 6 < 10 mmHg) was present in 30 (69.8%), and HVPG ≥ 10 mmHg in 13 (30.2%). The median CPA was 16% (IQR 10.75-23.25). Median Child-Pugh score and HVPG were not significantly different between Ishak fibrosis stage 5 or 6, whereas CPA was statistically different: 13% in stage 5 (IQR 8.3-12.4) versus 23% in stage 6 (IQR 17-33.7, P < 0.001). In the multivariate analysis, CPA was the only variable significantly associated with clinically-significant portal hypertension (HVPG ≥ 10 mmHg, odds ratio: 1.085, confidence interval: 1.004-1.172, P = 0.040). A CPA of 14% was the best cut-off value for clinically-significant portal hypertension (CSPH) and liver decompensation, which occurred in 24 patients. Event-free survival was significantly shorter in patients with CSPH or with a CPA value ≥ 14%, or with a combination of both. In Ishak stages 5 and 6, CPA correlated with HVPG, but had a wider range of values, suggesting a greater sensitivity for distinguishing "early" from "late" severe fibrosis/cirrhosis. CPA was a unique, independent predictor of HVPG ≥ 10 mmHg. CPA can be used to subclassify cirrhosis and for prognostic stratification.


Prevalence and Associated Factors of Fecal Incontinence and Double Incontinence among Rural Elderly in North China.
Fecal and double incontinence are known to be more prevalent among the rural elderly. Yet, there have been few studies on their epidemic condition among Chinese rural elders. This study estimated the prevalence and correlates of fecal and double incontinence in rural elderly aged 65 years and over in North China. A multisite cross-sectional survey was conducted in 10 villages, yielding a sampling frame of 1250 residents. Fecal and urinary incontinence assessments were based on the self-reported bowel health questionnaire and the International Consultation on Incontinence Questionnaire-Short Form, respectively. The concomitant presence of fecal and urinary incontinence in the same subject was defined as double incontinence. The prevalence of fecal and double incontinence was 12.3% and 9.3%, respectively. Factors associated with fecal incontinence included urinary incontinence, lack of social interaction, traumatic brain injury, cerebrovascular disease, and poverty. Physical activities of daily living dependence, traumatic brain injury, lack of social interaction, and poor sleep quality were associated with higher odds of having double incontinence, whereas tea consumption was correlated with lower odds. Individualized intervention programs should be developed targeting associated factors and high-risk populations. These intervention programs should be integrated into existing public health services for the rural elderly to facilitate appropriate prevention and management of incontinence.


Bacterial communities reflect the spatial variation in pollutant levels in Brazilian mangrove sediment.
The majority of oil from oceanic oil spills converges on coastal ecosystems such as mangrove forests. A major challenge to mangrove bioremediation is defining the mangrove's pollution levels and measuring its recuperation from pollution. Bioindicators can provide a welcome tool for defining such recovery. To determine if the microbial profiles reflected variation in the pollutants, samples from different locations within a single mangrove with a history of exposure to oil were chemically characterised, and the microbial populations were evaluated by a comprehensive range of conventional and molecular methods. Multivariate ordination of denaturing gradient gel electrophoresis (DGGE) microbial community fingerprints revealed a pronounced separation between the sediment and rhizosphere samples for all analysed bacterial communities (Bacteria, Betaproteobacteria, Alphaproteobacteria, Actinobacteria and Pseudomonas). A Mantel test revealed significant relationships between the sediment chemical fertility and oil-derived pollutants, most of the bacterial community fingerprints from sediment samples, and the counts by different cultivation strategies. The level of total petroleum hydrocarbons was significantly associated with the Bacteria and Betaproteobacteria fingerprints, whereas anthracene and the total level of polycyclic aromatic hydrocarbons were associated with the Actinobacteria. These results show that microbial communities from the studied mangrove reflect the spatial variation of the chemicals in the sediment, demonstrating the specific influences of oil-derived pollutants.


Cytotoxic Th1 cells mushroom after fungal exposures in Crohn's disease.
Fungi are consistently enriched in inflamed intestines, with elusive effects on host immunity. In a recent issue of Nature Medicine, Martini et al. identify a subset of Th1 cells able to lyse the epithelium, enriched in Crohn's disease patient samples after fungal exposure.


Overcoming the Challenges to Clinical Development of X-Linked Retinitis Pigmentosa Therapies: Proceedings of an Expert Panel.
X-linked retinitis pigmentosa (XLRP) is a rare inherited retinal disease manifesting as impaired night vision and peripheral vision loss that progresses to legal blindness. Although several trials of ocular gene therapy for XLRP have been conducted or are in progress, there is currently no approved treatment. In July 2022, the Foundation Fighting Blindness convened an expert panel to examine relevant research and make recommendations for overcoming the challenges and capitalizing on the opportunities in conducting clinical trials of RPGR-targeted therapy for XLRP. Data presented concerned RPGR structure and mutation types known to cause XLRP, RPGR mutation-associated retinal phenotype diversity, patterns in genotype/phenotype relationships, disease onset and progression from natural history studies, and the various functional and structural tests used to monitor disease progression. Panel recommendations include considerations, such as genetic screening and other factors that can impact clinical trial inclusion criteria, the influence of age on defining and stratifying participant cohorts, the importance of conducting natural history studies early in clinical development programs, and the merits and drawbacks of available tests for measuring treatment outcomes. We recognize the need to work with regulators to adopt clinically meaningful end points that would best determine the efficacy of a trial. Given the promise of RPGR-targeted gene therapy for XLRP and the difficulties encountered in phase III clinical trials to date, we hope these recommendations will help speed progress to finding a cure. Examination of relevant data and recommendations for the successful clinical development of gene therapies for RPGR-associated XLRP.


[Preliminary investigation on the formation mechanism of CCL4-water-cetyl trimethyl ammonium bromide (CTAB) gel].
Gels are gaining extensive interest owing to their versatile applications in fields such as drug delivery, tissue engineering, cosmetics, templated materials and food industry. Surfactants have an ability to self-assemble into a variety of supramolecular aggregate structures and morphologies. Of particular interest in resent years are surfactant-based gels, one special class of materials due to surfactant assemblies resulting in viscoelastic solid-like rheological behaviors. Up to now, there is only limited understanding on the mechanism of gel formation, especially on the interaction among water, organic solvents and surfactant during thegel formation. In this study we prepare a Low-molecule-gel that is composed of cetyl trimethyl ammonium bromide (CTAB), water and carbon tetrachloride. Based on the experimental result of XRD and titration, the authors find that CTAB in gel are more than in saturated CTAB solution but CTAB is not solide in gel. CTAB is not solvented in CCl4. The solubility of CTAB in saturated CTAB solution is limited. So the authors suppose that CTAB is a synergistically solubilized by water and CCl4 in the gel. In addition, both NMR and FTIR spectroscopic results demonstrate that CTAB cations form a quasi-ordered structure in the gel.


System, Method and Software for Calculation of a Cannabis Drug Efficiency Index for the Reduction of Inflammation.
There are many varieties of Cannabis sativa that differ from each other by composition of cannabinoids, terpenes and other molecules. The medicinal properties of these cultivars are often very different, with some being more efficient than others. This report describes the development of a method and software for the analysis of the efficiency of various cannabis extracts to detect the anti-inflammatory properties of the various cannabis extracts. The method uses high-throughput gene expression profiling data but can potentially use other omics data as well. According to the signaling pathway topology, the gene expression profiles are convoluted into the signaling pathway activities using a signaling pathway impact analysis (SPIA) method. The method was tested by inducing inflammation in human 3D epithelial tissues, including intestine, oral and skin, and then exposing these tissues to various extracts and then performing transcriptome analysis. The analysis showed a different efficiency of the various extracts in restoring the transcriptome changes to the pre-inflammation state, thus allowing to calculate a different cannabis drug efficiency index (CDEI).


Phosphoinositide 3-kinase regulates crosstalk between Trk A tyrosine kinase and p75(NTR)-dependent sphingolipid signaling pathways.
The mechanism of crosstalk between signaling pathways coupled to the Trk A and p75(NTR) neurotrophin receptors in PC12 cells was examined. In response to nerve growth factor (NGF), Trk A activation inhibited p75(NTR)-dependent sphingomyelin (SM) hydrolysis. The phosphoinositide 3-kinase (PI 3-kinase) inhibitor, LY294002, reversed this inhibition suggesting that Trk A activation of PI 3-kinase is necessary to inhibit sphingolipid signaling by p75(NTR). In contrast, SM hydrolysis induced by neurotrophin-3 (NT-3), which did not activate PI-3 kinase, was uneffected by LY294002. However, transient expression of a constituitively active PI 3-kinase inhibited p75(NTR)-dependent SM hydrolysis by both NGF and NT-3. Intriguingly, NGF induced an association of activated PI 3-kinase with acid sphingomyelinase (SMase). This interaction localized to caveolae-related domains and correlated with a 50% decrease in immunoprecipitated acid SMase activity. NGF-stimulated PI 3-kinase activity was necessary for inhibition of acid SMase but was not required for ligand-induced association of the p85 subunit of PI 3-kinase with the phospholipase. Finally, this interaction was specific for NGF since EGF did not induce an association of PI 3-kinase with acid SMase. In summary, our data suggest that PI 3-kinase regulates the inhibitory crosstalk between Trk A tyrosine kinase and p75(NTR)-dependent sphingolipid signaling pathways and that this interaction localizes to caveolae-related domains.


Oligonucleotide recombination in Gram-negative bacteria.
This report describes several key aspects of a novel form of RecA-independent homologous recombination. We found that synthetic single-stranded DNA oligonucleotides (oligos) introduced into bacteria by transformation can site-specifically recombine with bacterial chromosomes in the absence of any additional phage-encoded functions. Oligo recombination was tested in four genera of Gram-negative bacteria and in all cases evidence for recombination was apparent. The experiments presented here were designed with an eye towards learning to use oligo recombination in order to bootstrap identification and development of phage-encoded recombination systems for recombineering in a wide range of bacteria. The results show that oligo concentration and sequence have the greatest influence on recombination frequency, while oligo length was less important. Apart from the utility of oligo recombination, these findings also provide insights regarding the details of recombination mediated by phage-encoded functions. Establishing that oligos can recombine with bacterial genomes provides a link to similar observations of oligo recombination in archaea and eukaryotes suggesting the possibility that this process is evolutionary conserved.


The farnesyl transferase inhibitor (FTI) lonafarnib improves nuclear morphology in ZMPSTE24-deficient fibroblasts from patients with the progeroid disorder MAD-B.
Several related progeroid disorders are caused by defective post-translational processing of prelamin A, the precursor of the nuclear scaffold protein lamin A, encoded by LMNA. Prelamin A undergoes farnesylation and additional modifications at its C-terminus. Subsequently, the farnesylated C-terminal segment is cleaved off by the zinc metalloprotease ZMPSTE24. The premature aging disorder Hutchinson Gilford progeria syndrome (HGPS) and a related progeroid disease, mandibuloacral dysplasia (MAD-B), are caused by mutations in LMNA and ZMPSTE24, respectively, that result in failure to process the lamin A precursor and accumulate permanently farnesylated forms of prelamin A. The farnesyl transferase inhibitor (FTI) lonafarnib is known to correct the aberrant nuclear morphology of HGPS patient cells and improves lifespan in children with HGPS. Importantly, and in contrast to a previous report, we show here that FTI treatment also improves the aberrant nuclear phenotypes in MAD-B patient cells with mutations in ZMPSTE24 (P248L or L425P). As expected, lonafarnib does not correct nuclear defects for cells with lamin A processing-proficient mutations. We also examine prelamin A processing in fibroblasts from two individuals with a prevalent laminopathy mutation LMNA-R644C. Despite the proximity of residue R644 to the prelamin A cleavage site, neither R644C patient cell line shows a prelamin A processing defect, and both have normal nuclear morphology. This work clarifies the prelamin A processing status and role of FTIs in a variety of laminopathy patient cells and supports the FDA-approved indication for the FTI Zokinvy for patients with processing-deficient progeroid laminopathies, but not for patients with processing-proficient laminopathies.


Randomized, Placebo-Controlled, Double-Blind Phase 2 Trial Comparing the Reactogenicity and Immunogenicity of a Single Standard Dose to Those of a High Dose of CVD 103-HgR Live Attenuated Oral Cholera Vaccine, with Shanchol Inactivated Oral Vaccine as an Open-Label Immunologic Comparator.
Reactive immunization with a single-dose cholera vaccine that could rapidly (within days) protect immunologically naive individuals during virgin soil epidemics, when cholera reaches immunologically naive populations that have not experienced cholera for decades, would facilitate cholera control. One dose of attenuated Vibrio cholerae O1 classical Inaba vaccine CVD 103-HgR (Vaxchora) containing ≥2 × 108 CFU induces vibriocidal antibody seroconversion (a correlate of protection) in >90% of U.S. adults. A previous CVD 103-HgR commercial formulation required ≥2 × 109 CFU to elicit high levels of seroconversion in populations in developing countries. We compared the vibriocidal responses of Malians (individuals 18 to 45 years old) randomized to ingest a single ≥2 × 108-CFU standard dose (n = 50) or a ≥2 × 109-CFU high dose (n = 50) of PaxVax CVD 103-HgR with buffer or two doses (n = 50) of Shanchol inactivated cholera vaccine (the immunologic comparator). To maintain blinding, participants were dosed twice 2 weeks apart; CVD 103-HgR recipients ingested placebo 2 weeks before or after ingesting vaccine. Seroconversion (a ≥4-fold vibriocidal titer rise) between the baseline and 14 days after CVD 103-HgR ingestion and following the first and second doses of Shanchol were the main outcomes measured. By day 14 postvaccination, the rates of seroconversion after ingestion of a single standard dose and a high dose of CVD 103-HgR were 71.7% (33/46 participants) and 83.3% (40/48 participants), respectively. The rate of seroconversion following the first dose of Shanchol, 56.0% (28/50 participants), was significantly lower than that following the high dose of CVD 103-HgR (P = 0.003). The vibriocidal geometric mean titer (GMT) of the high dose of CVD 103-HgR exceeded the GMT of the standard dose at day 14 (214 versus 95, P = 0.045) and was ∼2-fold higher than the GMT on day 7 and day 14 following the first Shanchol dose (P > 0.05). High-dose CVD 103-HgR is recommended for accelerated evaluation in developing countries to assess its efficacy and practicality in field situations. (This study has been registered at ClinicalTrials.gov under registration no. NCT02145377.).


Neuroendoscopic management of symptomatic septum pellucidum cavum vergae cyst using a high-definition flexible endoscopic system.
A 24-year-old man, who had an asymptomatic septum pellucidum cyst incidentally found one year previously, presented with severe headache and right abducens nerve palsy caused by expansion of the midline cyst. Preoperative magnetic resonance (MR) imaging revealed obstructive hydrocephalus due to the enlarged midline cyst. Neuroendoscopic fenestration of the septum pellucidum cyst was successfully performed via a right frontal approach using a high-resolution flexible neuroendoscopic system without complication. Communication between the cyst cavity and bilateral lateral ventricles was constructed via a single trajectory. The entire inner cyst wall could be inspected from the cyst cavity by manipulating the flexible neuroendoscopic system, which excluded the presence of neoplasm. His symptoms were completely relieved after surgery, and postoperative MR imaging showed significant improvement of hydrocephalus and shrinkage of the midline cyst. Septum pellucidum cavum vergae cyst may expand and become symptomatic, so fenestration using a flexible neuroendoscope system may be the optimal method for constructing communication to the bilateral lateral ventricles with minimal invasion.


In vivo fluorescence of medullary carcinoma of the thyroid: a technology with potential to improve visualization of malignant tissue at surgical resection.
Medullary carcinoma of the thyroid requires aggressive treatment because of its potential to metastasize and because of the current limitations of preoperative localization and systemic therapy. If these tumors could be made to fluoresce in vivo with tagged fluorophore antibodies against tumor antigens, surgeons would be able to obtain additional information in the operating room to facilitate a more complete resection. Based on the success of our previous work in breast and colon cancer models, we conducted an animal study of in vivo tumor fluorescence of a human medullary thyroid cell line in which bright tumor fluorescence is visible during dissection. To accomplish this, we used an inexpensive and commercially available handheld, blue (470 nm), light-emitting diode flashlight and filtered goggles (520 nm). This procedure, which we call the fluorescent antibody-assisted surgical technique (FAAST), is easy to perform, requires no complex or expensive technical equipment, and has the potential to be applied to a wide variety of tumors. To the best of our knowledge, this is the first experiment of its kind to be reported in the literature.


Nucleotide proofreading functions by nematode RAD51 paralogs facilitate optimal RAD51 filament function.
The RAD51 recombinase assembles as helical nucleoprotein filaments on single-stranded DNA (ssDNA) and mediates invasion and strand exchange with homologous duplex DNA (dsDNA) during homologous recombination (HR), as well as protection and restart of stalled replication forks. Strand invasion by RAD51-ssDNA complexes depends on ATP binding. However, RAD51 can bind ssDNA in non-productive ADP-bound or nucleotide-free states, and ATP-RAD51-ssDNA complexes hydrolyse ATP over time. Here, we define unappreciated mechanisms by which the RAD51 paralog complex RFS-1/RIP-1 limits the accumulation of RAD-51-ssDNA complexes with unfavorable nucleotide content. We find RAD51 paralogs promote the turnover of ADP-bound RAD-51 from ssDNA, in striking contrast to their ability to stabilize productive ATP-bound RAD-51 nucleoprotein filaments. In addition, RFS-1/RIP-1 inhibits binding of nucleotide-free RAD-51 to ssDNA. We propose that 'nucleotide proofreading' activities of RAD51 paralogs co-operate to ensure the enrichment of active, ATP-bound RAD-51 filaments on ssDNA to promote HR.


Quantitative Proteomic Analysis of Staphylococcus aureus Treated With Punicalagin, a Natural Antibiotic From Pomegranate That Disrupts Iron Homeostasis and Induces SOS.
Staphylococcus aureus, a bacterial, food-borne pathogen of humans, can contaminate raw fruits and vegetables. While physical and chemical methods are available to control S. aureus, scientists are searching for inhibitory phytochemicals from plants. One promising compound from pomegranate is punicalagin, a natural antibiotic. To get a broader understanding of the inhibitory effect of punicalagin on S. aureus growth, high-throughput mass spectrometry and quantitative isobaric labeling was used to investigate the proteome of S. aureus after exposure to a sublethal dose of punicalagin. Nearly half of the proteins encoded by the small genome were interrogated, and nearly half of those exhibited significant changes in accumulation. Punicalagin treatment altered the accumulation of proteins and enzymes needed for iron acquisition, and it altered amounts of enzymes for glycolysis, citric acid cycling, protein biosynthesis, and purine and pyrimidine biosynthesis. Punicalagin treatment also induced an SOS cellular response to damaged DNA. Transcriptional comparison of marker genes shows that the punicalagin-induced iron starvation and SOS responses resembles those produced by EDTA and ciprofloxacin. These results show that punicalagin adversely alters bacterial growth by disrupting iron homeostasis and that it induces SOS, possibly through DNA biosynthesis inhibition.


Effect of different treatments on the activity of the heat labile enterotoxin of Escherichia coli F11(P155).
A whole cell lysate preparation of the Escherichia coli strain F11(P155) was treated with the following agents: heat (60 degrees C for 30 minutes), pronase, lipase, amylase, formalin (0.1%), sodium lauryl sulfate (0.05%) and sarkosyl NL30 (0.05%). Except for the amylase treatment, the treated whole cell lysate was inactivated when tested in rabbit gut loops or on Chinese hamster ovary cells. The preparations treated with heat or formalin could be used to produce a neutralizing antiserum or could remove the neutralizing capacity of an anti-F11(P155) serum. Finally, the attempt to demonstrate enterotoxicity with the cytoplasmic membrane of the bacterial cell failed.


Cigarette smoke induces cellular senescence.
Chronic obstructive pulmonary disease (COPD) is the fourth leading cause of death in the United States, and cigarette smoking is the major risk factor for COPD. Fibroblasts play an important role in repair and lung homeostasis. Recent studies have demonstrated a reduced growth rate for lung fibroblasts in patients with COPD. In this study we examined the effect of cigarette smoke extract (CSE) on fibroblast proliferative capacity. We found that cigarette smoke stopped proliferation of lung fibroblasts and upregulated two pathways linked to cell senescence (a biological process associated with cell longevity and an inability to replicate), p53 and p16-retinoblastoma protein pathways. We compared a single exposure of CSE to multiple exposures over an extended time course. A single exposure to CSE led to cell growth inhibition at multiple phases of the cell cycle without killing the cells. The decrease in proliferation was accompanied by increased ATM, p53, and p21 activity. However, several important senescent markers were not present in the cells at an earlier time point. When we examined multiple exposures to CSE, we found that the cells had profound growth arrest, a flat and enlarged morphology, upregulated p16, and senescence-associated beta-galactosidase activity, which is consistent with a classic senescent phenotype. These observations suggest that while a single exposure to cigarette smoke inhibits normal fibroblast proliferation (required for lung repair), multiple exposures to cigarette smoke move cells into an irreversible state of senescence. This inability to repair lung injury may be an essential feature of emphysema.


Conservative surgical management of malignant ovarian germ cell tumors: the experience of the Gynecologic Oncology Unit at Ain Shams University.
To evaluate the role of extended surgical staging in patients with malignant ovarian germ cell tumors in the presence of cisplatinum-based combination chemotherapy. 16 patients aged between 13 and 40 years (mean 20.5) diagnosed and treated for malignant ovarian germ cell tumors at the Gynecologic Oncology Unit. Ain Shams University, during the period from May 1994 to October 1999. Six patients were diagnosed with dysgerminoma, six with immature teratoma and four with endodermal sinus tumor. Only seven cases were primarily managed at the unit and were subjected to proper surgical staging as required by FIGO (two in each of the stages IC, IIC and IIIA and one in stage IV). However, the remaining nine cases were referred to the unit, six after having unilateral salpingo-oophorectomy and no surgical staging, one patient after total abdominal hysterectomy and bilateral salpingo-oophorectomy (TAH+BSO), and one with recurrent dysgerminoma in the retroperitoneum and mediastinum following suboptimal treatment. None of these cases were surgically re-explored and all including the first six cases were given the standard BEP chemotherapy for 4-6 courses (mean 5.8). Follow-up ranged from 7-72 months (mean 30.5). All patients are alive without any evidence of disease recurrence except for one patient with a stage IIIA immature teratoma who had a local and distant recurrence and is undergoing second-line chemotherapy. All patients have their menstrual function preserved except for three patients; one having 46,XX pure gonadal dysgenesis with a preoperative FSH level of 120 U/l and the eldest two patients having a TAH+BSO as they had completed their families. Three patients are currently pregnant. In view of the high chemosensitivity and curability of ovarian germ cell tumors and their occurrence in young patients, every effort should be made to preserve one ovary and the uterus for future reproduction even in advanced cases. The role of revisional surgical staging, in particular retroperitoneal lymphadenectomy in light of these data and those of others should be redefined in the future.


Aspects of regulation in kidney at the enzymic level: aldolase isozymes, aminopeptidase and alkaline phosphatase.
Histochemical and immuno-histochemical studies on kidney sections of adult rats and rats at different stages of development were carried out to estimate enzyme concentrations in nephron segments. Aminopeptidase and alkaline phosphatase were found from 3 days before birth in proximal tubule cells, aldolase-B and aldolase-A in all the nephron and collecting duct. The concentration of the enzymes remained remarkably constant from the 3rd day after birth onwards, except for aldolase-A in the distal tubule cells. The concentration of aldolase-B was higher than of aldolase-A in the distal tubule cells. The concentration of aldolase-B was higher than of aldolase-A, in glomerula and proximal tubules, that of aldolase-A was higher than that of aldolase-B in the ascending thick part of the Henle loop and in the following parts of the nephron. This implies that the concentration of the enzymes is the limiting factor for the regulation of substrate hydrolysis and that the nephron when formed can function as efficiently as the nephron of the adult rat with respect to these enzymes.


The effects of nanoparticle drug loading on the pharmacokinetics of anticancer agents.
Major advances in carrier-mediated agents, which include nanoparticles, nanosomes and conjugates, have revolutionized drug delivery capabilities over the past decade. While providing numerous advantages, such as greater solubility, duration of exposure and delivery to the site of action over their small-molecule counterparts, there is substantial variability in systemic clearance and distribution, tumor delivery and pharmacologic effects (efficacy and toxicity) of these agents. This review provides an overview of factors that affect the pharmacokinetics and pharmacodynamics of carrier-mediated agents in preclinical models and patients.


Effects of arm dominance and brachial artery cannulation on forearm blood flow measured by strain-gauge plethysmography.
The human forearm model is used extensively in physiological, pharmacological and clinical investigations. Effects of arm dominance or arterial cannulation on forearm flow measurements have never been tested formally. In the present study we tested the hypotheses that left or right arm dominance or cannulation of the brachial artery do not affect forearm haemodynamic responses to physiological or pharmacological stimuli. Results obtained in 16 volunteers showed that forearm blood flow responses to physiological stimuli are comparable before and after intra-arterial cannulation in either the dominant or the non-dominant forearm. Cannulation of a forearm brachial artery has a small effect on baseline blood flow. Responses to intra-arterially infused noradrenaline (norepinephrine) were not influenced by left or right arm dominance. Intravenous infusion of noradrenaline in eight subjects resulted in small responses in forearm blood flow that were slightly asymmetrical. During the intravenous infusion of noradrenaline, forearm blood flow or the forearm blood flow ratio did not reflect the marked increase in FVR that occurred. These results support our hypotheses (a) that either arm can be used as the control or intervention arm, and (b) that intra-arterial cannulation does not affect the results of intra-arterial infusion studies.


Randomized clinical trial of the effects of perioperative use of immune-enhancing enteral formula on metabolic and immunological status in patients undergoing esophagectomy.
Although perioperative immune-enhancing enteral formula (IEEF) is effective to decrease the rate of infectious complications, it is not clear whether perioperative use of IEEF decreases the incidence of postoperative complications and improves clinical outcome in patients who have undergone esophagectomy. A prospective randomized clinical trial was performed to examine the effects of perioperative IEEF on nutritional and immunological status in patients with esophageal carcinoma who have been treated with esophagectomy. A total of 30 patients were randomly assigned to two groups, each receiving 3 days of preoperative and postoperative enteral nutrition through jejunostomy started within 24 h after operation, either with immune-enhancing enteral formula (group IEEF, n = 16) or with regular polymeric enteral formula (group C, n = 14). Preoperative and postoperative nutritional and immunological parameters and clinical outcome were examined. A significant increase in the serum concentration of ornithine was noted in group IEEF and it peaked at 5 days after surgery. The equivalent values were significantly lower in group C. There was no difference in serum dochosahexaic acid between the two groups. The n-3/n-6 fatty acid ratio in group IEEF was significantly higher than in group C at 7 days after surgery. Peripheral percent lymphocyte fraction and total lymphocyte count in group IEEF were both significantly higher than those in group C. While T cell fraction of peripheral lymphocytes in group IEEF at 3 days after surgery, B cell fraction in group IEEF at 5 and 7 days after surgery was significantly higher than those in group C, suggesting that perioperative IEEF caused a shift towards B cell proliferation. Perioperative use of IEEF caused a significant increase in the total lymphocyte count at 3 and 5 days after operation and caused a shift toward B cell proliferation, which may possibly be beneficial to decrease the incidence of postoperative infectious complications.


The relationship of inferior ST depression, lateral ST elevation, and left precordial ST elevation to myocardium at risk in acute anterior myocardial infarction.
The purpose of this study was to examine the relationship between the presence or absence of ST segment depression in inferior leads (II, III, and aVF) and ST segment elevation in lateral (I and aVL) or left precordial (V5 and V6) leads with the amount and location of myocardium at risk for infarction in patients with acute anterior myocardial infarction. Forty-three patients with anterior infarctions were injected with technetium 99m-sestamibi when they were first seen and underwent tomographic imaging to measure the amount and location of myocardium at risk. Patients with inferior ST depression (n = 10) compared with those without ST depression (n = 33) had perfusion defects that extended significantly further into the lateral wall (47 degrees vs 20 degrees, p = 0.04) and larger anterior injury vectors (6.47 vs 4.92, p = 0.008). There was no significant association with the percentage of myocardium at risk, disease of the right coronary artery, the presence of an inferior perfusion defect, or the size of the inferior injury vector. Among the patients with ST elevation in lateral leads (n = 16) compared with those without (n = 27), there was a significantly more lateral defect border (47 degrees vs 25 degrees, p = 0.007) and a larger anterior injury vector (6.07 vs 4.81, p = 0.01). There was no significant correlation with the percentage of myocardium at risk. A significant relationship could not be demonstrated between the presence of ST elevation in the left precordial leads and any measure of the amount or location of myocardium at risk. These data support the theory that inferior ST depression in patients with transmural anterior ischemia is a "reciprocal" finding and does not represent inferior ischemia. The presence of inferior ST depression or lateral ST elevation is associated with a more lateral perfusion defect. Neither of these ECG findings is associated with the amount of myocardium at risk for infarction.


Influenza virus NS vectors expressing the mycobacterium tuberculosis ESAT-6 protein induce CD4+ Th1 immune response and protect animals against tuberculosis challenge.
Infection with Mycobacterium tuberculosis remains a major cause of morbidity and mortality all over the world. Since the effectiveness of the only available tuberculosis vaccine, Mycobacterium bovis bacillus Calmette-Guérin (BCG), is suboptimal, there is a strong demand to develop new tuberculosis vaccines. As tuberculosis is an airborne disease, the intranasal route of vaccination might be preferable. Live influenza virus vaccines might be considered as potential vectors for mucosal immunization against various viral or bacterial pathogens, including M. tuberculosis. We generated several subtypes of attenuated recombinant influenza A viruses expressing the 6-kDa early secretory antigenic target protein (ESAT-6) of M. tuberculosis from the NS1 reading frame. We were able to demonstrate the potency of influenza virus NS vectors to induce an M. tuberculosis-specific Th1 immune response in mice. Moreover, intranasal immunization of mice and guinea pigs with such vectors induced protection against mycobacterial challenge, similar to that induced by BCG vaccination.


Prevalence and risk of malignancy of focal incidental uptake detected by fluorine-18-fluorodeoxyglucose positron emission tomography in the parotid gland: a meta-analysis.
This study aimed at performing a meta-analysis on the prevalence and risk of malignancy of focal parotid incidental uptake (FPIU) detected by hybrid fluorine-18-fluorodeoxyglucose ((18)F-FDG) positron emission tomography/computed tomography (PET/CT) or (18)F-FDG PET alone. A comprehensive literature search of studies published up to July 2014 was performed. Records reporting at least 5 FPIUs were selected. Pooled prevalence and malignancy risk of FPIU were calculated including 95 % confidence intervals (95 % CI). Twelve records were selected for our meta-analysis. Pooled prevalence of FPIU detected by (18)F-FDG PET or PET/CT was 0.6 % (95 % CI 0.4-0.7 %), collecting data of 220 patients with FPIU. Overall, 181 FPIUs underwent further evaluation and 165 FPIUs were pathologically proven. Pooled risk of malignancy was 9.6 % (95 % CI 5.4-14.8 %), 10.9 % (95 % CI 5.8-17.3 %) and 20.4 % (95 % CI 12.3-30 %), considering all FPIUs detected, only those which underwent further evaluation and only those pathologically proven, respectively. Selection bias in the included studies, the heterogeneity among studies and the publication bias are limitations of our meta-analysis. Overall FPIUs are observed in about 1 % of (18)F-FDG PET or PET/CT scans and they are benign in most of the cases. Nevertheless, further evaluation is needed whenever FPIUs are detected by (18)F-FDG-PET or PET/CT to exclude malignant lesions or with possible malignant degeneration. Prospective studies are needed to confirm the findings reported by our meta-analysis.


Diagnostic accuracy of the Heidelberg Retina Tomograph for glaucoma a population-based assessment.
To evaluate the Heidelberg Retina Tomograph II (HRTII; Heidelberg Engineering, Heidelberg, Germany) as a screening tool for open-angle glaucoma (OAG) in an older population. Population-based, cross-sectional study. Participants from the 10-year follow-up Blue Mountains Eye Study (n = 1952; 75.6% of survivors). Participants underwent optic nerve head imaging performed using the HRTII; OAG was diagnosed independently from optic disc photographs and Humphrey 24-2 visual fields. The Moorfields Regression Analysis (MRA) was applied to scans using the default results of "normal," borderline," and "outside normal limits." Outcome classification was at the person level and used data from both eyes. Mean age was 73.7 years. The HRT scans could be acquired in 1644 participants, 95.9% of those fully examined; 87.4% of scans having a topography standard deviation (TSD) < or =40 microm. Larger TSD was associated with older age and OAG. The MRA sensitivity was 64.1%, specificity 85.7%, positive predictive value 21%, and negative predictive value 97.6% for detecting OAG. Including borderline results improved sensitivity (87.0%) but specificity dropped to 70.6%. Significant predictors of abnormal MRA included OAG (odds ratio [OR], 7.70; 95% confidence interval [CI] 4.79-12.35), age (OR, 1.05; 95% CI, 1.03-1.07), TSD (OR, 1.01; 95% CI, 1.003-1.020) and disc size (OR, 3.2; 95% CI, 2.31-4.49). Diagnostic accuracy was improved by restricting TSD <40 microm. Further TSD restriction improved specificity at the expense of sensitivity. As disc size increased, specificity fell whereas sensitivity, OAG prevalence, and the proportion testing positive rose. Glaucoma prevalence, positive predictive value, and the proportion testing positive increased, but specificity fell with increasing age. Sensitivity also fell between youngest and oldest groups. Diagnostic performance improved for visual field mean deviation < -4.0 dB. Single eye analyses overestimated specificity and underestimated sensitivity compared with using data from both eyes. Diagnostic test evaluation studies of HRTII using single eye analyses or restrictive selection overestimate test accuracy compared with this population-based study. Although the specificity of the MRA was inadequate for use as a glaucoma screening test, the HRTII performed relatively well in an unselected older population with acceptable quality scans in most eyes.


Broad spectrum infrared thermal desorption of wipe-based explosive and narcotic samples for trace mass spectrometric detection.
Wipe collected analytes were thermally desorbed using broad spectrum near infrared heating for mass spectrometric detection. Employing a twin tube filament-based infrared emitter, rapid and efficiently powered thermal desorption and detection of nanogram levels of explosives and narcotics was demonstrated. The infrared thermal desorption (IRTD) platform developed here used multi-mode heating (direct radiation and secondary conduction from substrate and subsequent convection from air) and a temperature ramp to efficiently desorb analytes with vapor pressures across eight orders of magnitude. The wipe substrate experienced heating rates up to (85 ± 2) °C s-1 with a time constant of (3.9 ± 0.2) s for 100% power emission. The detection of trace analytes was also demonstrated from complex mixtures, including plastic-bonded explosives and exogenous narcotics, explosives, and metabolites from collected artificial latent fingerprints. Manipulation of the emission power and duration directly controlled the heating rate and maximum temperature, enabling differential thermal desorption and a level of upstream separation for enhanced specificity. Transitioning from 100% power and 5 s emission duration to 25% power and 30 s emission enabled an order of magnitude increase in the temporal separation (single seconds to tens of seconds) of the desorption of volatile and semi-volatile species within a collected fingerprint. This mode of operation reduced local gas-phase concentrations, reducing matrix effects experienced with high concentration mixtures. IRTD provides a unique platform for the desorption of trace analytes from wipe collections, an area of importance to the security sector, transportation agencies, and customs and border protection.


Mitogenic stimulation of resting T cells causes rapid phosphorylation of the transcription factor LSF and increased DNA-binding activity.
The mammalian transcription factor LSF (CP2/LBP-1c) binds cellular promoters modulated by cell growth signals. We demonstrate here that LSF-DNA-binding activity is strikingly regulated by induction of cell growth in human peripheral T lymphocytes. Within 15 min of mitogenic stimulation of these cells, the level of LSF-DNA-binding activity increased by a factor of five. The level of LSF protein in the nucleus remained constant throughout this interval. However, a rapid decrease in the electrophoretic mobility of LSF, attributable to phosphorylation, correlated with the increase in DNA-binding activity. pp44 (ERK1) phosphorylated LSF in vitro on the same residue that was phosphorylated in vivo, specifically at amino acid position 291, as indicated by mutant analysis. As direct verification of the causal relationship between phosphorylation and DNA-binding activity, treatment in vitro of LSF with phosphatase both increased the electrophoretic mobility of the protein and decreased LSF-DNA-binding activity. This modulation of LSF-DNA-binding activity as T cells progress from a resting to a replicating state reveals that LSF activity is regulated during cell growth and suggests that LSF regulates growth-responsive promoters.


Coexistence of Lipid Phases Stabilizes Interstitial Water in the Outer Layer of Mammalian Skin.
The lipid matrix in the outer layer of mammalian skin, the stratum corneum, has been previously investigated by multiple biophysical techniques aimed at identifying hydrophilic and lipophilic pathways of permeation. Although consensus is developing over the microscopic structure of the lipid matrix, no molecular-resolution model describes the permeability of all chemical species simultaneously. Using molecular dynamics simulations of a model mixture of skin lipids, the self-assembly of the lipid matrix lamellae has been studied. At higher humidity, the resulting lamellar phase is maintained by partitioning excess water into isolated droplets of controlled size and spatial distribution. The droplets may fuse together to form intralamellar water channels, thereby providing a pathway for the permeation of hydrophilic species. These results reconcile competing data on the outer skin's structure and broaden the scope of molecular-based methods to improve the safety of topical products and to advance transdermal drug delivery.


Cerebral torque is human specific and unrelated to brain size.
The term "cerebral torque" refers to opposing right-left asymmetries of frontal and parieto-occipital regions. These are assumed to derive from a lateralized gradient of embryological development of the human brain. To establish the timing of its evolution, we computed and compared the torque, in terms of three principal features, namely petalia, shift, and bending of the inter-hemispheric fissure as well as the inter-hemispheric asymmetry of brain length, height and width for in vivo Magnetic Resonance Imaging (MRI) scans of 91 human and 78 chimpanzee brains. We found that the cerebral torque is specific to the human brain and that its magnitude is independent of brain size and that it comprises features that are inter-related. These findings are consistent with the concept that a "punctuational" genetic change of relatively large effect introduced lateralization in the hominid lineage. The existence of the cerebral torque remains an unsolved mystery and the present study provides further support for this most prominent structural brain asymmetry being specific to the human brain. Establishing the genetic origins of the torque may, therefore, have relevance for a better understanding on human evolution, the organisation of the human brain, and, perhaps, also aspects of the neural basis of language.


[Iron deficiency anaemia--interpretation of biochemical and haematological findings].
Iron deficiency and iron deficiency anaemia are frequent problems in both the primary and the specialist health services. It is important to detect iron deficiency and to determine the causal relationship because iron deficiency may be secondary to a serious disease. The diagnosis of iron deficiency is largely based on biochemical and haematological laboratory findings, but there is no standardisation or consensus on the interpretation of these findings. Non-systematic search in the PubMed database with a discretionary selection of articles, based on the authors' knowledge of the field. Ferritin measurement is the most important analysis in the study of iron deficiency, but there is no consensus on the diagnostic cut-off. It is usual in Norway today to use a ferritin level of < 12-20 μg/L, but at this low level the sensitivity for detecting iron deficiency is very low. A number of studies show that if the diagnostic cut-off is increased to the order of 30 μg/L the sensitivity is significantly higher for only a small reduction in specificity. When studying iron deficiency as a cause of anaemia, the diagnostic cut-off for detecting deficiency should be higher than that used today. The ferritin level increases with inflammation and ought in practice to be considered in conjunction with the CRP level. The level of transferrin receptor in plasma increases with iron deficiency without being influenced by inflammation and is therefore a good supplement to ferritin measurement. Measurement of iron, transferrin and transferrin saturation provides little information additional to that provided by ferritin in iron deficiency studies.


Prevalence and incidence of osteoporotic fractures in patients on long-term glucocorticoid treatment for rheumatic diseases: the Glucocorticoid Induced OsTeoporosis TOol (GIOTTO) study.
Osteoporosis and fractures are common and invalidating consequences of chronic glucorticoid (GC) treatment. Reliable information regarding the epidemiology of GC induced osteoporosis (GIOP) comes exclusively from the placebo group of randomized clinical trials while observational studies are generally lacking data on the real prevalence of vertebral fractures, GC dosage and primary diagnosis. The objective of this study was to evaluate the prevalence and incidence of osteoporotic fractures and to identify their major determinants (primary disease, GC dosage, bone mineral density, risk factors, specific treatment for GIOP) in a large cohort of consecutive patients aged >21 years, on chronic treatment with GC (≥5 mg prednisone - PN - equivalent) and attending rheumatology centers located all over Italy. Glucocorticoid Induced OsTeoporosis TOol (GIOTTO) is a national multicenter cross-sectional and longitudinal observational study. 553 patients suffering from Rheumatoid Arthritis (RA), Polymyalgia Rheumatica (PMR) and Connective Tissue Diseases (CTDs) and in chronic treatment with GCs were enrolled. Osteoporotic BMD values (T score <-2.5) were observed in 28%, 38% and 35% of patients with CTDs, PMR or RA at the lumbar spine, and in 18%, 29% and 26% at the femoral neck, respectively. Before GC treatment, prevalent clinical fractures were reported by 12%, 37% and 17% of patients with CTDs, PMR, or RA, respectively. New clinical fragility fractures during GC treatment were reported by 12%, 10% and 23% of CTDs, PMR and RA patients, respectively. Vertebral fractures were the prevailing type of fragility fracture. More than 30% of patients had recurrence of fracture. An average of 80% of patients were in supplementation with calcium and/or vitamin D during treatment with GCs. Respectively, 64%, 80%, and 72% of the CTDs, PMR and RA patients were on pharmacological treatment for GIOP, almost exclusively with bisphosphonates. The GIOTTO study might provide relevant contributions to clinical practice, in particular by highlighting and quantifying in real life the prevalence of GIOP and relative fractures, the frequency of the main risk factors, and the currently sub-optimal prevention. Moreover, these results emphasize the importance of the underlying rheumatic disease on the risk of GIOP associated fractures.


The prevalence of neck pain in migraine.
To determine the prevalence of neck pain at the time of migraine treatment relative to the prevalence of nausea, a defining associated symptom of migraine. This is a prospective, observational cross-sectional study of 113 migraineurs, ranging in attack frequency from episodic to chronic migraine. Subjects were examined by headache medicine specialists to confirm the diagnosis of migraine and exclude both cervicogenic headache and fibromyalgia. Details of all migraines were recorded over the course of at least 1 month and until 6 qualifying migraines had been treated. For each attack, subjects recorded the presence or absence of nausea as well as the intensity of headache and neck pain (graded as none, mild, moderate, or severe). Subjects recorded 2411 headache days, 786 of which were migraines. The majority of migraines were treated in the moderate pain stage. Regardless of the intensity of headache pain at time of treatment, neck pain was a more frequent accompaniment of migraine than was nausea (P< .0001). Prevalence of neck pain correlated with chronicity of headache as attacks moved from episodic to chronic daily headache. In this representative cross-section of migraineurs, neck pain was more commonly associated with migraine than was nausea, a defining characteristic of the disorder. Awareness of neck pain as a common associated feature of migraine may improve diagnostic accuracy and have a beneficial impact on time to treatment.


Roles of NAD+ in Health and Aging.
NAD+, the essential metabolite involved in multiple reactions such as the regulation of cellular metabolism, energy production, DNA repair, mitophagy and autophagy, inflammation, and neuronal function, has been the subject of intense research in the field of aging and disease over the last decade. NAD+ levels decline with aging and in some age-related diseases, and reduction in NAD+ affects all the hallmarks of aging. Here, we present an overview of the discovery of NAD+, the cellular pathways of producing and consuming NAD+, and discuss how imbalances in the production rate and cellular request of NAD+ likely contribute to aging and age-related diseases including neurodegeneration. Preclinical studies have revealed great potential for NAD+ precursors in promotion of healthy aging and improvement of neurodegeneration. This has led to the initiation of several clinical trials with NAD+ precursors to treat accelerated aging, age-associated dysfunctions, and diseases including Alzheimer's and Parkinson's. NAD supplementation has great future potential clinically, and these studies will also provide insight into the mechanisms of aging.


Acyclovir treatment of disseminated varicella in childhood malignant neoplasms.
Primary varicella-zoster virus infection (chickenpox) in immunocompromised children is frequently associated with visceral dissemination and attendant high mortality. Eight children with malignant neoplasms and chickenpox with visceral involvement (seven with hepatitis, three with pneumonitis, two with encephalitis, and two with coagulopathy) were initially treated with intravenously (IV) administered vidarabine but demonstrated progressive visceral involvement. After three days of vidarabine treatment (two days for two patients), seven had rising serum SGPT levels, all eight had pneumonitis, seven had deteriorating mental status and/or seizure activity, and six had worsening coagulopathy. Vidarabine was replaced by IV administered acyclovir, with subsequent improvement in all but the most severely ill patient who died. Seven of eight patients recovered completely; no side effects of acyclovir were observed. This clinical experience suggests that acyclovir may be more effective than vidarabine in disseminated varicella infection; however, controlled clinical trials will be necessary to establish this.


A Decade of Improvement in Door-to-Needle Time Among Acute Ischemic Stroke Patients, 2008 to 2017.
The clinical benefit of intravenous (IV) alteplase in acute ischemic stroke is time dependent. We assessed the overall temporal changes in door-to-needle (DTN) time and examine the factors associated with DTN time ≤60 and ≤45 minutes. A total of 496 336 acute ischemic stroke admissions were identified in the Paul Coverdell National Acute Stroke Program from 2008 to 2017. We used generalized estimating equations models to examine the factors associated with DTN time ≤60 and ≤45 minutes, and calculated adjusted odds ratios and 95% CI. Between 2008 and 2017, the percentage of acute ischemic stroke patients who received IV alteplase including those transferred, increased from 6.4% to 15.3%. After excluding those who received IV alteplase at an outside hospital, a total of 39 737 (8%) acute ischemic stroke patients received IV alteplase within 4.5 hours of the time the patient last known to be well. Significant increases were seen in DTN time ≤60 minutes (26.4% in 2008 to 66.2% in 2017, P<0.001), as well as DTN time ≤45 minutes (10.7% in 2008 to 40.5% in 2017, P<0.001). Patients aged 55 to 84 years were more likely to receive IV alteplase within 60 minutes, while those aged 55 to 74 years were more likely to receive IV alteplase within 45 minutes, as compared with those aged 18 to 54 years. Arrival by emergency medical service, and patients with severe stroke were more likely to receive IV alteplase within 60 and 45 minutes. Conversely, women, black patients as compared with white, and patients with a medical history of diseases associated with stroke were less likely to receive DTN time ≤60 or 45 minutes. Rapid improvements in DTN time were observed in the Paul Coverdell National Acute Stroke Program; however, opportunities to reduce disparities remain.


Depressive symptoms are the strongest predictors of short-term declines in health status in patients with heart failure.
The purpose of this study was to assess whether depressive symptoms are independently associated with changes in heart failure (HF)-specific health status. Depression is common in patients with HF, but the impact of depressive symptoms on the health status of these patients over time is unknown. We conducted a multicenter prospective cohort study of outpatients with HF. Data from 460 patients who completed a baseline Medical Outcomes Study-Depression Questionnaire and both a baseline and follow-up (6 +/- 2 weeks) Kansas City Cardiomyopathy Questionnaire (KCCQ) were analyzed. The KCCQ measures HF-specific health status, including symptoms, physical and social function, and quality of life. Multivariable regression was used to evaluate depressive symptoms as a predictor of change in KCCQ scores, adjusting for baseline KCCQ scores and other patient variables. The primary outcome was change in KCCQ summary scores (range 0 to 100; higher scores indicate better health status; 5 points is a clinically meaningful change). Approximately 30% (139/460) of the patients had significant depressive symptoms at baseline. Depressed patients had markedly lower baseline KCCQ summary scores (beta = -19.6; p < 0.001). After adjustment for potential confounders, depressed patients were at risk for significant worsening of their HF symptoms, physical and social function, and quality of life (average change in KCCQ summary score = -7.1 points; p < 0.001). Depressive symptoms were the strongest predictor of decline in health status in the multivariable models. Depressive symptoms are a strong predictor of short-term worsening of HF-specific health status. The recognition and treatment of depression may be an important component of HF care.


The Updated NICE Guidelines: Cardiac CT as the First-Line Test for Coronary Artery Disease.
Cost-effective care pathways are integral to delivering sustainable healthcare programmes. Due to the overestimation of coronary artery disease using traditional risk tables, non-invasive testing has been utilised to improve risk stratification and initiate appropriate management to reduce the dependence on invasive investigations. In line with recent technological improvements, cardiac CT is a modality that offers a detailed anatomical assessment of coronary artery disease comparable to invasive coronary angiography. The recent publication of the National Institute for Health and Care Excellences (NICE) Clinical Guideline 95 update assesses the performance and cost utility of different non-invasive imaging strategies in patients presenting with suspected anginal chest pain. The low cost and high sensitivity of cardiac CT makes it the non-invasive test of choice in the evaluation of stable angina. This has now been ratified in national guidelines with NICE recommending cardiac CT as the first-line investigation for all patients presenting with chest pain due to suspected coronary artery disease. Additionally, randomised controlled trials have demonstrated that cardiac CT improves diagnostic certainty when incorporated into chest pain pathways. NICE recommend cardiac CT as the first-line test for the evaluation of stable coronary artery disease in chest pain pathways.


Crystal structure of 4-chlorocatechol 1,2-dioxygenase from the chlorophenol-utilizing gram-positive Rhodococcus opacus 1CP.
The crystal structure of the 4-chlorocatechol 1,2-dioxygenase from the Gram-positive bacterium Rhodococcus opacus (erythropolis) 1CP, a Fe(III) ion-containing enzyme involved in the aerobic biodegradation of chloroaromatic compounds, has been solved by multiple wavelength anomalous dispersion using the weak anomalous signal of the two catalytic irons (1 Fe/257 amino acids) and refined at a 2.5 A resolution (R(free) 28.7%; R factor 21.4%). The analysis of the structure and its comparison with the structure of catechol 1,2-dioxygenase from Acinetobacter calcoaceticus ADP1 (Ac 1,2-CTD) highlight significant differences between these enzymes. The general topology of the present enzyme comprises two catalytic domains (one for each subunit) related by a noncrystallographic 2-fold axis and separated by a common alpha-helical zipper motif consisting of five N-terminal helices from each subunit; furthermore the C-terminal tail is shortened significantly with respect to the known Ac 1,2-CTD. The presence of two phospholipids binding in a hydrophobic tunnel along the dimer axis is shown here to be a common feature for this class of enzyme. The active site cavity presents several dissimilarities with respect to the known catechol-cleaving enzyme. The catalytic nonheme iron(III) ion is bound to the side chains of Tyr-134, Tyr-169, His-194, and His-196, and a cocrystallized benzoate ion, bound to the metal center, reveals details on a novel mode of binding of bidentate inhibitors and a distinctive hydrogen bond network with the surrounding ligands. Among the amino acid residues expected to interact with substrates, several are different from the corresponding analogs of Ac 1,2-CTD: Asp-52, Ala-53, Gly-76, Phe-78, and Cys-224; in addition, regions of largely conserved amino acid residues in the catalytic cleft show different shapes resulting from several substantial backbone and side chain shifts. The present structure is the first of intradiol dioxygenases that specifically catalyze the cleavage of chlorocatechols, key intermediates in the aerobic catabolism of toxic chloroaromatics.


Creation of Architecturally Minimal Transcriptionally Activating Riboswitches Responsive to Theophylline Reveals an Unconventional Design Strategy.
Riboswitches are noncoding RNA switches that are largely utilized in bacteria and play a significant role in synthetic biology. Nonetheless, their natural counterparts possess lengthy sequences and intricate structures, posing challenges for their modular integration into complex gene circuits. Consequently, it is imperative to develop simplified synthetic riboswitches that can be effortlessly incorporated into gene circuits. The conventional approach to generate synthetic riboswitches entails tedious library construction and extensive screening, which frequently yields suboptimal performance. To overcome this obstacle, alternative methods are urgently needed. In this study, we created a novel approach to designing a diverse set of transcription-activating riboswitches that exhibit high performance and broad compatibility. The strategy involved starting with a synthetic theophylline RNA aptamer and designing an expression platform that forms a transcriptional terminator in its inactive state but switches to an antiterminator when it is activated. Several sequences were designed, constructed, and subjected to virtual screening, resulting in the identification of two transcription-activating riboswitches. These riboswitches were then engineered to reduce the basal leakage and increase the activation level through extending the hairpin region using a screened random sequence. These architecturally minimal synthetic riboswitches were highly adapted to different constitutive promoters in a modular manner, generating a differentially responsive output to theophylline. As a proof-of-principle, the synthetic riboswitches were applied to rewire a synthetic quorum-sensing circuit (QSC). The reprogrammed QSC successfully modulated the temporal responsive profile against the activation. This strategy is expected to expand the variety of high-performance riboswitches that are responsive to different ligands, thereby further facilitating the design of complex genetic circuits.


Influence of bonding agent application method on the dentin bond durability of a two-step adhesive utilizing a universal-adhesive-derived primer.
This study investigated the effect of the bonding agent application method on the dentin bond durability of a two-step adhesive utilizing a universal-adhesive-derived primer. The bonding durability of a universal adhesive was compared with those of two conventional two-step adhesives by shear bond strength testing after thermal cycling. The primer was applied (with or without phosphoric acid pre-etching), and the specimens were divided into three groups based on the bonding agent application method used as follows: (i) strong air-flow application, (ii) gentle air-flow application, and (iii) gentle air-flow + second bonding application. The shear bond strength was determined after thermal cycling. All the factors (bonding agent application method, adhesive system, and storage condition) significantly influenced the dentin shear bond strength both with and without phosphoric acid pre-etching. The specimens exposed to gentle air-flow showed a higher shear bond strength than did those exposed to strong air-flow and gentle air-flow + second bonding. The bond durability of the tested adhesive systems was influenced by the bonding agent application method, and this trend was material dependent. The G2-Bond Universal exhibited the same or greater dentin bond durability to the other two-step adhesive systems.


Amphetamine makes caudate tissue more susceptible to oxygen and glucose deprivation.
Amphetamine is being investigated to reduce morbidity following stroke. However, the medicinal use of amphetamine is complicated because this drug is addictive, cardiotoxic, and can be neurotoxic. Thus, further research into the safety of giving amphetamine to stroke patients is required. Here, we examine whether prior treatment with amphetamine has any effect on oxygen-glucose deprivation (OGD)-evoked transmitter efflux and mitochondrial function. To circumvent the well-documented cardiovascular effects of amphetamine we have used rat brain slices. Brain slices were exposed to 30μM of amphetamine for 10min 1h before being exposed to OGD. Using fast cyclic voltammetry in rat caudate, dopamine efflux induced by OGD was measured. The effect of amphetamine on mitochondrial function was examined using triphenyltetrazolium chloride (TTC) staining. Prior amphetamine exposure decreased the time to onset of OGD-evoked dopamine efflux (from 460 to 220s), suggesting that the caudate was more sensitive to OGD. This increased sensitivity to OGD was attenuated by pre-treatment with the dopamine transporter blocker GBR12909 (1μM). Pre-treatment with the dopamine D2 receptor antagonist metoclopramide (1μM) had no effect on the amphetamine-evoked sensitisation to OGD. Amphetamine decreased TTC staining in the caudate suggesting that amphetamine compromised the dopamine system by disrupting mitochondrial function. Amphetamine treatment may be harmful in stroke recovery by making the brain more vulnerable to ischaemia. These data also suggest that amphetamine abusers might be more susceptible to cerebral ischaemia.


Frequencies of emergency department use and hospitalization comparing patients with different types of substance or polysubstance-related disorders.
This study measured emergency department (ED) use and hospitalization for medical reasons among patients with substance-related disorders (SRD), comparing four subgroups: cannabis-related disorders, drug-related disorders other than cannabis, alcohol-related disorders and polysubstance-related disorders, controlling for various clinical, sociodemographic and service use variables. Clinical administrative data for a cohort of 22,484 patients registered in Quebec (Canada) addiction treatment centers in 2012-13 were extracted for the years 2009-10 to 2015-16. Using negative binomial models, risks of frequent ED use and hospitalization were calculated for a 12-month period (2015-16). Patients with polysubstance-related disorders used ED more frequently than other groups with SRD. They were hospitalized more frequently than patients with cannabis or other drug-related disorders, but less frequently than those with alcohol-related disorders. Patients with alcohol-related disorders used ED more frequently than those with cannabis-related disorders and underwent more hospitalizations than both patients with cannabis-related and other drug-related disorders. Co-occurring SRD-mental disorders or SRD-chronic physical illnesses, more years with SRD, being women, living in rural territories, more frequent consultations with usual general practitioner or outpatient psychiatrist, and receiving more interventions in community healthcare centers increased frequency of ED use and hospitalization, whereas both adverse outcomes decreased with high continuity of physician care. Behavioral addiction, age less than 45 years, living in more materially deprived areas, and receiving 1-3 interventions in addiction treatment centers increased risk of frequent ED use, whereas living in semi-urban areas decreased ED use. Patients 25-44 years old receiving 4+ interventions in addiction treatment centers experienced less frequent hospitalization. Findings showed higher risk of ED use among patients with polysubstance-related disorders, and higher hospitalization risk among patients with alcohol-related disorders, compared with patients affected by cannabis and other drug-related disorders. However, other variables contributed substantially more to the frequency of ED use and hospitalization, particularly clinical variables regarding complexity and severity of health conditions, followed by service use variables. Another important finding was that high continuity of physician care helped decrease the use of acute care services. Strategies like integrated care and outreach interventions may enhance SRD services.


The p53/miR-145a Axis Promotes Cellular Senescence and Inhibits Osteogenic Differentiation by Targeting Cbfb in Mesenchymal Stem Cells.
The osteogenic differentiation capacity of senescent bone marrow mesenchymal stem cells (MSCs) is reduced. p53 not only regulates cellular senescence but also functions as a negative regulator in bone formation. However, the role of p53 in MSCs senescence and differentiation has not been extensively explored. In the present study, we investigated the molecular mechanism of p53 in MSCs senescence and osteogenic differentiation. We found that p53 was upregulated during cellular senescence and osteogenic differentiation of MSCs respectively induced by H2O2 and BMP9. Similarly, the expression of p53-induced miR-145a was increased significantly. Furthermore, Overexpression of miR-145a in MSCs promoted cellular senescence and inhibited osteogenic differentiation. Then, we identified that p53-induced miR-145a inhibited osteogenic differentiation by targeting core binding factor beta (Cbfb), and the restoration of Cbfb expression rescued the inhibitory effects of miRNA-145a. In summary, our results indicate that p53/miR-145a axis exert its functions both in promoting senescence and inhibiting osteogenesis of MSCs, and the novel p53/miR-145a/Cbfb axis in osteogenic differentiation of MSCs may represent new targets in the treatment of osteoporosis.


Late recurrence and progression in non-muscle-invasive bladder cancers after 5-year tumor-free periods.
To evaluate the recurrence and progression in patients with non-muscle-invasive bladder tumors who remained tumor-free for at least 5 years, which should assist in the development of schedules of their follow-up evaluations. Non-muscle-invasive bladder tumors that recur or progress at a late time point are not rare. Between 1985 and 2002, 814 cases diagnosed with non-muscle-invasive bladder cancer were treated with transurethral resection. Of these 814 cases, 262 patients with no tumor recurrence for more than 5 years were included in the study. The median follow-up interval was 10.0 years. During the follow-up period, 39 tumors (14.9%) showed tumor recurrence. The 5- and 10-year recurrence-free survival rates were 81.6% and 76.0%, respectively. There was no significant difference in tumor recurrence among the low-, intermediate-, and high-risk groups based on the current clinical guideline. Only the use of intravesical mitomycin C was determined to be a significant unfavorable risk factor for late recurrence. Five patients (1.9%) experienced stage progression, 3 of whom did not have metastases at the time of diagnosis of the progression but died because of bladder cancer disease. After a 5-year tumor-free period, even in the low-risk group, recurrence occurred at a late time point to a degree that was the same as that for the intermediate- and high-risk groups. Finally, some of the high-risk patients experienced late progression with a high degree of malignant behavior, suggesting longer follow-up is needed for each patient.


Brain function and conditioning in posttraumatic stress disorder.
Posttraumatic stress disorder (PTSD) is commonly treated by psychotherapy, which may draw upon behavioural psychology or cognitive-behavioural psychology, thereby making use of desensitisation techniques--amongst others; hypnotherapy may also be used. Hypnotherapy and psychotherapy are also available for helping patients who suffer from symptoms associated with general stress or who show phobic symptoms, such as a fear of heights or of walking across bridges. Studies of patients with such disorders have not always linked emotional (affective) and behavioural symptoms with psychophysical factors, which correlate with the symptoms. The present article not only does this, but also shows that it is possible for brain function and psychoemotional outcome to mislead a person and 'trick' him or her into believing that certain fears or panics appear 'out of the blue' or might be due to experiences other than the true cause. These may be important when, for instance, childhood memories form an issue in any court case.


Perspectives in Breast and Ovarian Cancer Chemotherapy by Nanomedicine Approach: Nanoformulations in Clinical Research.
Breast and ovarian carcinomas represent major health problems in women worldwide. Chemotherapy constitutes the main treatment strategy, and the use of nanocarriers, a good tool to improve it. Several nanoformulations have already been approved, and others are under clinical trials for the treatment of both types of cancers. This review focuses on the analysis of the nanoformulations that are under clinical research in the treatment of these neoplasms. Currently, there are 6 nanoformulations in clinical trials for breast and ovarian carcinomas, most of them in phase II and phase III. In the case of breast cancer treatment, these nanomedicines contain paclitaxel; and, for ovarian cancer, nanoformulations containing paclitaxel or camptothecin analogs are being evaluated. The nanoencapsulation of these antineoplastics facilitates their administration and reduces their systemic toxicity. Nevertheless, the final approval and commercialization of nanoformulations may be limited by other aspects like lack of correlation between the efficacy results evaluated at in vitro and in vivo levels, difficulty in producing large batches of nanoformulations in a reproducible manner and high production costs compared to conventional formulations of antineoplastics. However, these challenges are not insurmountable and the number of approved nanoformulations for cancer therapy is growing. Reviewed nanoformulations have shown, in general, excellent results, demonstrating a good safety profile, a higher maximum tolerated dose and a similar or even slightly better antitumor efficacy compared to the administration of free drugs, reinforcing the use of nano-chemotherapy in both breast and ovarian tumors.


Estimation of potential soil erosion in the Prosecco DOCG area (NE Italy), toward a soil footprint of bottled sparkling wine production in different land-management scenarios.
Agricultural lands are the widest Human-modified ecosystems, making crop production the most extensive form of land use on Earth. However, in conventional agricultural land management, soil erosion may be boosted up to 1-2 orders of magnitude higher than the natural rates of soil production, making unproductive about the 30% of the world's arable. Nowadays in Europe, vineyards represent the most erosion-prone agricultural lands, especially in Mediterranean countries, showing the highest erosion rates in comparison to other type of land uses. Prosecco wine is produced in NE Italy by a rate of 400 M bottles per year, with the fastest growing demand in the global market at present. A production of 90 M bottles year-1 is currently running in the historical Prosecco DOCG (215 km2), in a steep hilly landscape of Veneto Region (Conegliano-Valdobbiadene). To sustain wine production, agricultural intensification is at present increasing, by re-setting of hillslopes and land use changes towards new vineyard plantations. The aim of this study is to estimate and to map potential soil erosion rate, calculating a sort of "soil footprint" for wine production in different agricultural land-management scenarios. RUSLE model was adopted to estimate potential soil erosion in Mg ha-1 year-1, by using high resolution topographic data (LiDAR), 10 years rainfall data analysis, detailed land use and local soil characteristics. For a conventional land-management scenario the estimated that total potential soil erosion in the Prosecco DOCG area is 411,266 Mg year-1, with an erosion rate of 19.5 Mg ha year-1. Modelled soil erosion is mainly clustered on steep slopes, with rates higher than 40 Mg ha-1 year-1. In Prosecco vineyards potential soil erosion could reach 300,180 Mg year-1, by a mean rate of 43.7 Mg ha-1 year-1, which is 31 times higher than the upper limit of tolerable soil erosion threshold defined for Europe. In contrast, simulation of different nature-based scenarios (hedgerows, buffer strips, and grass cover) showed soil erosion could be effectively reduced: a 100% inter-row grass cover showed a reduction of almost 3 times in vineyards (from 43.7 to 14.6 Mg ha-1 year-1), saving about 50% of soil in the whole Prosecco DOCG. The soil footprint modelled for a conventional land-management scenario is about 3.3 kg every bottle produced; in contrast it would be reduced to 1.1 kg/bottle in the completely green land-management scenario. This study, as the first estimation of potential soil erosion at Prosecco DOCG scale, suggests that an integrated and public soil erosion monitoring system is strongly needed in viticultural area, by implementing direct/indirect field measures with spatial analyses at agricultural landscape scale.


Short-Term Efficacy and Safety of Cataract Surgery Combined with Iris-Fixated Phakic Intraocular Lens Explantation: A Multicentre Study.
The purpose of this study was to evaluate the short-term efficacy and safety of cataract surgery for patients with iris-fixated phakic intraocular lenses (pIOLs). This study included 96 eyes of 91 patients. The changes in the logMAR uncorrected visual acuity (UCVA), best-corrected visual acuity (BCVA), subjective spherical equivalent (SE), astigmatism, and endothelial cell density (ECD) were collected retrospectively. The intraoperative and postoperative complications also were investigated to assess the surgical safety. The preoperative UCVA and BCVA improved significantly at month 1 postoperatively, respectively (p < 0.001 for both comparisons). The efficacy and safety index at month 1 postoperatively were 1.02 ± 0.56 and 1.31 ± 0.64, respectively. The SE at month 1 postoperatively was significantly (p < 0.001) higher compared to preoperatively, whereas the subjective astigmatism did not differ significantly (p = 0.078). The ECD significantly decreased at month 1 (p < 0.001). The most common postoperative complication was intraocular pressure elevation exceeding 25 mmHg in 10.4% of eyes, which was controlled with medications in all cases until month 1 postoperatively. No intraoperative complications developed. Cataract surgeries for patients with iris-fixated pIOLs were performed safely with good visual outcomes.


Gasdermin D inhibition prevents multiple organ dysfunction during sepsis by blocking NET formation.
Multiple organ dysfunction is the most severe outcome of sepsis progression and is highly correlated with a worse prognosis. Excessive neutrophil extracellular traps (NETs) are critical players in the development of organ failure during sepsis. Therefore, interventions targeting NET release would likely effectively prevent NET-based organ injury associated with this disease. Herein, we demonstrate that the pore-forming protein gasdermin D (GSDMD) is active in neutrophils from septic humans and mice and plays a crucial role in NET release. Inhibition of GSDMD with disulfiram or genic deletion abrogated NET formation, reducing multiple organ dysfunction and sepsis lethality. Mechanistically, we demonstrate that during sepsis, activation of the caspase-11/GSDMD pathway controls NET release by neutrophils during sepsis. In summary, our findings uncover a novel therapeutic use for disulfiram and suggest that GSDMD is a therapeutic target to improve sepsis treatment.


August Rauber (1841-1917): from the primitive streak to Cellularmechanik.
In the early 19th century Karl Ernst von Baer initiated a new research program searching for the mechanisms by which an egg transforms itself into an embryo. August Rauber (1841-1917) took up this challenge. He considered the phylogenetic principle as the right tool to explain the similitude of embryogenetic processes. In extending Baer's approach, he combined comparative embryology and histology in his studies of avian and mammalian embryos. His earlier work demonstrated that the two-layered chick embryo is a modified gastrula and not a "disc" as Wilhelm His had claimed. From the 1880s onwards, he concentrated on the issue of how the development of germ layers is related to tissue differentiation. To address this, he studied the blastopore, epiblast, primitive streak, teratology and the relative importance of nucleus and cytoplasm in heredity. This paper reconstructs some of Rauber's work and concludes that his observations and reflections constituted a new approach combining embryology and histology with "phylogenetic" reasoning.


Self-driven microstructural evolution of Au@Pd core-shell nanoparticles for greatly enhanced catalytic performance during methanol electrooxidation.
The lack of direct insight into the microstructural evolution of catalytic materials under electrochemical polarization has inhibited the development of heterogeneous catalysts. By investigating a typical Au@Pd core-shell nanostructure, the present study discloses the microstructural evolution of heterogeneous catalytic materials during the methanol electrooxidation reaction (MOR). The electrocatalytic activity of the as-prepared Au@Pd_core-shell nanoparticles continuously increased during the first 100 successive voltammetry cycles of the MOR. Microstructural characterization studies revealed that during the MOR, an Au/Pd mixed bimetallic shell was formed by the self-driven microstructural evolution of the Au@Pd_core-shell nanoparticles. Both the experimental and calculation results indicated that the Au/Pd mixed bimetallic shell reduced the binding strength of OH- and CO on the catalyst surface. The exposed Au atoms in the shell region also produced large-scale reactive ˙OH radicals that facilitated the oxidative removal of the adsorbed carbonaceous species from the adjacent Pd active sites.


Preferences towards novel foods in Cebus apella: the role of nutrients and social influences.
Information on the process of preference acquisition towards novel foods in nonhuman primates is lacking. This study aims to assess (1) whether nutrient and energy contents affect preferences towards novel foods encountered repeatedly by individuals when alone, (2) whether these preferences change after additional encounters with the novel foods, and (3) if the change is sensitive to social influences. We presented seven novel foods to 26 socially housed tufted capuchins. In Phase 1, each subject was presented individually with the 21 possible binary combinations of the seven novel foods. Afterwards, during treatment, 13 subjects received the novel foods ad libitum with their group members (social condition) and 13 subjects received the foods individually (individual condition); subjects assigned to the individual and social conditions had shown similar food preferences in Phase 1. Finally, in Phase 2, each subject was presented again with 21 binary choices between each of the novel foods. In Phases 1 and 2, the number of times each food was chosen differed among foods. In Phase 1, food preference correlated positively with glucose and fructose and negatively with total fiber content. In Phase 2, irrespective of social or individual prior experience in the treatment condition, food preference changed and became correlated with total energy content. Our results broaden the findings already available for familiar foods by demonstrating that individual experience based on the feedback obtained from novel foods guides the establishment of preferences towards them. Moreover, individual experience is sufficient to determine food preferences similar to those individuals may acquire when together with group members eating the same foods.


Experiences of outdoor nature-based therapeutic recreation programs for persons with a mental illness: a qualitative systematic review.
The objective of this review was to identify, appraise, and synthesize the best available evidence related to participation in outdoor nature-based therapeutic recreation programs for adults with a mental illness living in the community. Therapeutic recreation is posited to be beneficial for persons living with a mental illness. Research evidence indicates that therapeutic recreation programs can foster mental health recovery. It is important to understand the effectiveness of this from the perspective of persons living with mental illness how outdoor nature-based therapeutic recreation programs are meaningful and helpful for recovery. This review considered studies that collected qualitative data on the experiences and perspectives of adults with a mental illness regarding their participation in outdoor nature-based therapeutic recreation programs. The databases PsycINFO, CINAHL, MEDLINE, Scopus, and Informit, as well as unpublished sources in gray literature (Google Scholar, OpenGrey), were searched and reference lists were checked to locate any additional studies. Studies published in English were considered, with a date range from inception to 2019. Three reviewers independently assessed the methodological quality of the studies that met the inclusion criteria using the JBI critical appraisal approach for qualitative research. Data were extracted by one reviewer using the standardized qualitative extraction tool and checked for accuracy by three other reviewers. The qualitative research findings were pooled using JBI methodology. The JBI process of meta-aggregation was used to identify categories and synthesized findings, and a level of confidence was assigned to both synthesized findings. Eighteen papers met the inclusion criteria and were included in the data extraction phase. A total of 84 findings were extracted and aggregated into six categories based on similarity of meaning and two synthesized findings. The methodological quality of the studies varied, and the overall level of confidence of the synthesized findings was determined to be moderate. This review identified that persons living with mental illness perceive outdoor nature-based therapeutic recreation as enjoyable and that therapeutic recreation makes a positive contribution to mental health. Congruent with the literature, therapeutic recreation offers a socially inclusive and psychologically safe environment. The intentionally structured social milieu enhances the formation of social relationships and meaningful connections for persons with mental illness. Elements linked with psychological well-being, such as intrinsic motivation, overcoming perceived challenges, and finding purpose and meaning, are enhanced through participation in therapeutic recreation in outdoor nature-based settings. Increased levels of physical activity, greater self-esteem, and enhanced sense of identity were some of the perceived positive changes. This review provides important insights into the subjective needs of persons with mental illness who undertake therapeutic recreation in outdoor nature-based settings. The qualitative findings can inform health care providers, or those interested in therapeutic recreation programming, to use alongside quantitative evidence of effectiveness to design nature-based therapeutic recreation activities that are meaningful for persons with mental illness. Limitations of the research were that papers published in languages other than English were not searched, and papers not located may have influenced the findings of this review.


Characterization of genetic diversity and population structure within Staphylococcus chromogenes by multilocus sequence typing.
Staphylococcus chromogenes is a common skin commensal in cattle and has been identified as a frequent cause of bovine mastitis and intramammary infections. We have developed a seven locus Multilocus Sequence Typing (MLST) scheme for typing S. chromogenes. Sequence-based typing systems, such as MLST, have application in studies of genetic diversity, population structure, and epidemiology, including studies of strain variation as a factor in pathogenicity or host adaptation. The S. chromogenes scheme was tested on 120 isolates collected from three geographic locations, Vermont and Washington State in the United States and Belgium. A total of 46 sequence types (STs) were identified with most of the STs being location specific. The utility of the typing scheme is indicated by a discrimination power of 95.6% for all isolates and greater than 90% for isolates from each of the three locations. Phylogenetic analysis placed 39 of the 46 STs into single core group consistent with a common genetic lineage; the STs in this group differ by less than 0.5% at the nucleotide sequence level. Most of the diversification in this lineage group can be attributed to mutation; recombination plays a limited role. This lineage group includes two clusters of single nucleotide variants in starburst configurations indicative of recent clonal expansion; nearly 50% of the isolates sampled in this study are in these two clusters. The remaining seven STs were set apart from the core group by having alleles with highly variable sequences at one or more loci. Recombination had a higher impact than mutation in the diversification of these outlier STs. Alleles with hypervariable sequences were detected at five of the seven loci used in the MLST scheme; the average sequence distances between the hypervariable alleles and the common core alleles ranged from 12 to 34 nucleotides. The extent of these sequence differences suggests the hypervariable alleles may be remnants of an ancestral genotype.