ZAP-70 compared with immunoglobulin heavy-chain gene mutation status as a predictor of disease progression in chronic lymphocytic leukemia.
The course of chronic lymphocytic leukemia (CLL) is variable. In aggressive disease, the CLL cells usually express an unmutated immunoglobulin heavy-chain variable-region gene (IgV(H)) and the 70-kD zeta-associated protein (ZAP-70), whereas in indolent disease, the CLL cells usually express mutated IgV(H) but lack expression of ZAP-70. We evaluated the CLL B cells from 307 patients with CLL for ZAP-70 and mutations in the rearranged IgV(H) gene. We then investigated the association between the results and the time from diagnosis to initial therapy. We found that ZAP-70 was expressed above a defined threshold level in 117 of the 164 patients with an unmutated IgV(H) gene (71 percent), but in only 24 of the 143 patients with a mutated IgV(H) gene (17 percent, P<0.001). Among the patients with ZAP-70-positive CLL cells, the median time from diagnosis to initial therapy in those who had an unmutated IgV(H) gene (2.8 years) was not significantly different from the median time in those who had a mutated IgV(H) gene (4.2 years, P=0.07). However, the median time from diagnosis to initial treatment in each of these groups was significantly shorter than the time in patients with ZAP-70-negative CLL cells who had either mutated or unmutated IgV(H) genes (P<0.001). The median time from diagnosis to initial therapy among patients who did not have ZAP-70 was 11.0 years in those with a mutated IgV(H) gene and 7.1 years in those with an unmutated IgV(H) gene (P<0.001). Although the presence of an unmutated IgV(H) gene is strongly associated with the expression of ZAP-70, ZAP-70 is a stronger predictor of the need for treatment in B-cell CLL.


Collagen copolymer toric posterior chamber phakic intraocular lens in eyes with keratoconus.
To assess the safety, efficacy, stability, and predictability of collagen copolymer toric phakic intraocular lens (pIOL) implantation to correct myopia and astigmatism in eyes with keratoconus. Fernández-Vega Ophthalmological Institute, Oviedo, Spain. This prospective study comprised keratoconic eyes that had implantation of a toric Intraocular Collamer Lens. Uncorrected (UDVA) and corrected (CDVA) distance visual acuities, refraction, and postoperative complications were evaluated 1, 3, 6, and 12 months postoperatively. Preoperatively, the mean spherical equivalent in the 30 eyes (21 patients) was -5.38 diopters (D) +/- 3.26 (SD) (range -13.50 to -0.63 D) and the mean cylinder, -3.48 +/- 1.24 D (range -1.75 to -6.00 D). At 12 months, 86.7% of the eyes were within +/-0.50 D of the attempted refraction and all eyes were within +/-1.00 D. For the astigmatic components J0 and J45, 83.3% of eyes and 86.7% of eyes, respectively, were within +/-0.50 D. The mean Snellen UDVA was 0.81 +/- 0.20 and the mean CDVA, 0.83 +/- 0.18; CDVA was 20/40 or better in 29 eyes 96.7% of eyes and 20/25 or better in 22 eyes (73.3%). No eyes lost more than 2 lines of CDVA; 29 eyes (96.7%) maintained or gained 1 or more lines. The efficacy index was 1.07 and the safety index, 1.16. There were no complications or adverse events. The results confirm that toric ICL implantation is a predictable, effective procedure to correct ametropia in eyes with keratoconus. Predictability and stability were achieved early and remained fairly stable up to 12 months.


MEFV gene variations in patients with systemic lupus erythematosus.
The aim of this study was to investigate the frequency of familial Mediterranean fever (FMF)-associated MEFV gene variations in patients with systemic lupus erythematosus (SLE). The study group comprised 190 SLE patients and 101 healthy controls of Turkish origin with no clinical features of FMF. All individuals were genotyped for the four most common MEFV gene variations (M694V, M680I, V726A and E148Q) by PCR-restriction fragment length polymorphism analysis. The frequency of carrying any of the four MEFV gene variations under study was 15 % in patients with SLE and 10 % in the healthy controls (p = 0.23). After the exclusion of the less penetrant E148Q variation, re-analysis for the three penetrant mutations revealed a significant association between exon 10 variations and pericarditis [p = 0.038, odds ratio (OR) 3.5, 95 % confidence interval (CI) 1.0-12.1], and pleural effusion (p = 0.043, OR 5.2, 95 % CI 0.8-30.9). No significant association was detected between the MEFV gene variations and a higher acute phase response. The MEFV gene variations analyzed in our study do not seem to increase the overall susceptibility to SLE and do not have any strong association with its clinical manifestations. The possibility of a modest effect of penetrant exon 10 MEFV variants on the development of serosal effusions needs to be explored in a larger series of patients.


Aphanomyces euteiches cell wall fractions containing novel glucan-chitosaccharides induce defense genes and nuclear calcium oscillations in the plant host Medicago truncatula.
N-acetylglucosamine-based saccharides (chitosaccharides) are components of microbial cell walls and act as molecular signals during host-microbe interactions. In the legume plant Medicago truncatula, the perception of lipochitooligosaccharide signals produced by symbiotic rhizobia and arbuscular mycorrhizal fungi involves the Nod Factor Perception (NFP) lysin motif receptor-like protein and leads to the activation of the so-called common symbiotic pathway. In rice and Arabidopsis, lysin motif receptors are involved in the perception of chitooligosaccharides released by pathogenic fungi, resulting in the activation of plant immunity. Here we report the structural characterization of atypical chitosaccharides from the oomycete pathogen Aphanomyces euteiches, and their biological activity on the host Medicago truncatula. Using a combination of biochemical and biophysical approaches, we show that these chitosaccharides are linked to β-1,6-glucans, and contain a β-(1,3;1,4)-glucan backbone whose β-1,3-linked glucose units are substituted on their C-6 carbon by either glucose or N-acetylglucosamine residues. This is the first description of this type of structural motif in eukaryotic cell walls. Glucan-chitosaccharide fractions of A. euteiches induced the expression of defense marker genes in Medicago truncatula seedlings independently from the presence of a functional Nod Factor Perception protein. Furthermore, one of the glucan-chitosaccharide fractions elicited calcium oscillations in the nucleus of root cells. In contrast to the asymmetric oscillatory calcium spiking induced by symbiotic lipochitooligosaccharides, this response depends neither on the Nod Factor Perception protein nor on the common symbiotic pathway. These findings open new perspectives in oomycete cell wall biology and elicitor recognition and signaling in legumes.


Constitutional chromothripsis rearrangements involve clustered double-stranded DNA breaks and nonhomologous repair mechanisms.
Chromothripsis represents a novel phenomenon in the structural variation landscape of cancer genomes. Here, we analyze the genomes of ten patients with congenital disease who were preselected to carry complex chromosomal rearrangements with more than two breakpoints. The rearrangements displayed unanticipated complexity resembling chromothripsis. We find that eight of them contain hallmarks of multiple clustered double-stranded DNA breaks (DSBs) on one or more chromosomes. In addition, nucleotide resolution analysis of 98 breakpoint junctions indicates that break repair involves nonhomologous or microhomology-mediated end joining. We observed that these eight rearrangements are balanced or contain sporadic deletions ranging in size between a few hundred base pairs and several megabases. The two remaining complex rearrangements did not display signs of DSBs and contain duplications, indicative of rearrangement processes involving template switching. Our work provides detailed insight into the characteristics of chromothripsis and supports a role for clustered DSBs driving some constitutional chromothripsis rearrangements.


Catecholaminergic polymorphic ventricular tachycardia (and seizure) caused by a novel homozygous likely pathogenic variant in CASQ2 gene.
Although structural heart disease is frequently present among patients who experience sudden cardiac death (SCD), inherited arrhythmia syndromes can also play an important role in the occurrence of SCD. CPVT2, which is the second-most prevalent form of CPVT, arises from an abnormality in the CASQ2 gene. We represent a novel CASQ2 variant that causes CPVT2 and conduct a comprehensive review on this topic. The proband underwent Whole-exome sequencing (WES) in order to ascertain the etiology of CPVT. Subsequently, the process of segregating the available family members was carried out through the utilization of PCR and Sanger Sequencing. We searched the google scholar and PubMed/Medline for studies reporting CASQ2 variants, published up to May 10,2023. We used the following mesh term "Calsequestrin" and using free-text method with terms including "CASQ2","CASQ2 variants", and "CASQ2 mutation". The CASQ2 gene was found to contain an autosomal recessive nonsense variant c.268_269insTA:p.Gly90ValfsTer4, which was identified by WES. This variant was determined to be the most probable cause of CPVT in the pedigree under investigation. CASQ2 variants play an important role in pathogenesis of CPVT2. Notabely, based on results of our study and other findings in the literature the variant in this gene may cause an neurological signs in the patients with CPVT2. Further studies are needed for more details about the role of this gene in CPVT evaluation, diagnosis, and gene therapy.


Early stutterings: some aspects of their form and distribution.
The speech of five stutterers ranging in age from 3 years, 10 months to 5 years, 7 months was analyzed to determine the types and loci of stutterings. Word repetition was the most frequent feature in three cases and one of the two predominant features in the remaining two cases. Almost without exception, word repetitions occurred at the beginning of syntactic units. A greater proportion of stutterings of all types appeared on the initial words of sentences or clauses than on the other words. In most cases proportionately more function words than control words were stuttered, as were more monosyllabic than polysyllabic words--just the reverse of the usual pattern in older children and adults. The tendency of older stutterers to have more difficulty on initial consonants than initial vowels appeared in only one case. The findings on the properties of stuttered words were interpreted to mean that word-bound factors as such have little influence on the loci of early stutterings. The results as a whole were related to the hypothesis that early stuttering represents mainly a type of difficulty in either the formulation or the execution of syntactic units.


Mitochondrial N-formylmethionyl proteins as chemoattractants for neutrophils.
Mitochondria synthesize several hydrophobic proteins. Like bacteria, mitochondria initiate protein synthesis with an N-formylmethionine residue. Because N-formylmethionyl peptides have been found to be chemotactic for polymorphonuclear leukocytes (PMN), mitochondria isolated from cultured human cells and purified bovine mitochondrial proteins were tested for PMN chemotactic activity in vitro. Nondisrupted mitochondria were not chemotactic. However, intact mitochondria that had been incubated with a lysosomal lysate did stimulate PMN migration. Antibodies directed against two mitochondrial enzymes, cytochrome oxidase and ATPase, (both of which contain mitochondrially synthesized subunits) but not anti-C3 or anti-C5 decreased mitochondrially derived chemotactic activity. In addition, purified bovine mitochondrial N-formylmethionyl proteins stimulated PMN migration in vitro, whereas nonformylated mitochondrial proteins did not. Furthermore, the chemotactic activity of purified mitochondrial proteins and disrupted mitochondria was decreased by the formyl peptide antagonist butyloxycarbonyl-phenylalanine-leucine-phenylalanine-leucine-phenylalanine. Finally, disrupted mitochondria and purified mitochondrial proteins stimulated PMN-directed migration (chemotaxis), according to accepted criteria. In addition to other chemotactic factors, release of N-formylmethionyl proteins from mitochondria at sites of tissue damage, may play a role in the accumulation of inflammatory cells at these sites.


Cystic nephroma: a histologic and immunohistochemical study of 10 cases.
Cystic nephroma is a rare and controversial benign multicystic renal tumor. While the clinical, radiologic, and histologic features of cystic nephroma are well described, the immunohistochemical features are not. The role of immunohistochemistry in the differential diagnosis, which includes multicystic renal cell carcinoma, is also unknown. To define the histologic and immunohistochemical features of cystic nephroma. Ten cases of cystic nephroma diagnosed at 2 institutions during a period of 10 years were stained with an immunohistochemical panel consisting of 20 immunostains. Median age at diagnosis was 61 years, with a range from 31 to 79 years. The female-to-male ratio was 9:1. Grossly, the tumors were multicystic masses without solid nodules. Histologic features included cysts lined by flat, cuboidal, or hobnail epithelium and septa variably lined by fibrous (10/10 cases) and/or ovarian-like (7/10 cases) stroma. Corpus albicans-like acellular hyalinized structures were noted in the septa in 9 of 10 cases. The cyst epithelium showed consistent positivity for distal tubule/collecting duct markers (cytokeratin 19, cytokeratin AE1/AE3, epithelial membrane antigen) and variable positivity for proximal tubule markers (alpha1-antitrypsin, lysozyme, CD15, CD10). The ovarian-like stroma (present in 7/10 cases) stained positively for progesterone receptors (6/7 cases) and estrogen receptors (4/7 cases). Our immunohistochemical findings confirm a previous report of both distal tubule/collecting duct and proximal tubule differentiation in cystic nephroma. Stromal estrogen and/or progesterone receptor positivity in the majority of cases of cystic nephroma is a novel finding.


The intrinsic vasculature of developing vertebral end plates and its nutritive significance to the intervertebral discs.
The structure and anatomical relationships of cartilage canals in the developing vertebral endplates of rabbits and humans are described in this study. In newborn rabbits, we observed a similar morphology and topologic arrangement to that in human fetal spines. Each vascular organ consisted of an arteriole, a glomerular tuft of sinusoidal capillaries, recurrent venules, and supporting loose connective tissue. Because the canals terminate adjacent to the intervertebral discs, they may provide an important source of nutrition to the disc during development when it is sandwiched between two layers of thick hyaline cartilage and contains no intrinsic vascular supply.


[Mobilization transfer of the pUB110 plasmid between gram-positive bacteria].
The three factor crosses between the donor strain Bacillus subtilis 168 harbouring the plasmid pUB102-4, Bacillus thuringiensis strain carrying the mobilizing plasmid pAM beta 1 and recipient strain Lactobacillus fermenti were conducted in order to elaborate the optimal conditions of the plasmid pUB102-4 mobilization for transfer into gram-positive microorganisms and to elucidate the possible expression of endogluconase genes in a lactobacillus strain. The Lactobacillus fermenti transconjugants carrying the pUB102-4 plasmid were obtained in the three factor reciprocal crosses with the streptococcus recipient strain and Bacillus subtilis recipients. The presence of the plasmids in transconjugants was confirmed by colony hybridization with the [32P]-labelled plasmid DNA and KMC-ase activity in transconjugant cells. The proposed system of crosses using the high copy number plasmid derivatives of pUB110 mobilized with high frequency by the pAM beta 1 plasmid demonstrates the possibility to increase the circle of gram-positive host bacteria avoiding time and labour consuming operations.


Association Between the Release of Netflix's 13 Reasons Why and Suicide Rates in the United States: An Interrupted Time Series Analysis.
To estimate the association between the release of the Netflix series 13 Reasons Why and suicide rates in the United States. Using segmented quasi-Poisson regression and Holt-Winters forecasting models, we assessed monthly rates of suicide among individuals aged 10 to 64 years grouped into 3 age categories (10-17, 18-29, and 30-64 years) between January 1, 2013, and December 31, 2017, before and after the release of 13 Reasons Why on March 31, 2017. We also assessed the impact of the show's release on a control outcome, homicide deaths. After accounting for seasonal effects and an underlying increasing trend in monthly suicide rates, the overall suicide rate among 10- to 17-year-olds increased significantly in the month immediately following the release of 13 Reasons Why (incidence rate ratio [IRR], 1.29; 95% CI, 1.09-1.53); Holt-Winters forecasting revealed elevated observed suicide rates in the month after release and in two subsequent months, relative to corresponding forecasted rates. Contrary to expectations, these associations were restricted to boys. Among 18- to 29-year-olds and 30- to 64-year-olds, we found no significant change in level or trend of suicide after the show's release, both overall and by sex. The show's release had no apparent impact in the control analyses of homicide deaths within any age group. The release of 13 Reasons Why was associated with a significant increase in monthly suicide rates among US youth aged 10 to 17 years. Caution regarding the exposure of children and adolescents to the series is warranted.


Immediate coronary angiography in survivors of out-of-hospital cardiac arrest.
The incidence of acute coronary-artery occlusion among patients with sudden cardiac arrest outside of the hospital is unknown, and the role of reperfusion therapy has not been determined. We therefore performed immediate coronary angiography and angioplasty when indicated in survivors of out-of-hospital cardiac arrest. Between September 1994 and August 1996, coronary angiography was performed in 84 consecutive patients between the ages of 30 and 75 years who had no obvious noncardiac cause of cardiac arrest. Sixty of the 84 patients had clinically significant coronary disease on angiography, 40 of whom had coronary-artery occlusion (48 percent). Angioplasty was attempted in 37 patients and was technically successful in 28. Clinical and electrocardiographic findings, such as the occurrence of chest pain and the presence of ST-segment elevation, were poor predictors of acute coronary-artery occlusion. The in-hospital survival rate was 38 percent. Multivariate logistic-regression analysis revealed that successful angioplasty was an independent predictor of survival (odds ratio, 5.2; 95 percent confidence interval, 1.1 to 24.5; P=0.04). Acute coronary-artery occlusion is frequent in survivors of out-of-hospital cardiac arrest and is predicted poorly by clinical and electrocardiographic findings. Accurate diagnosis by immediate coronary angiography can be followed in suitable candidates by coronary angioplasty, which seems to improve survival.


Post-transcriptional regulation of ornithine decarboxylase in Xenopus laevis oocytes.
The level at which ornithine decarboxylase expression is regulated in growing oocytes has been investigated. Immunoprecipitation of the in vivo labelled proteins showed that ornithine decarboxylase accumulated less rapidly in stage IV oocytes than in previtellogenic stage I + II oocytes. Quantitative Northern analysis showed that ornithine decarboxylase mRNA is abundant in oocytes (about 8 x 10(8) transcripts/cell) and this number does not significantly change during oogenesis. Polysome analysis showed that this mRNA is present in polysomes in stage I + II oocytes but has passed into puromycin-insensitive mRNP particles by stage IV of oogenesis. Therefore, during the growth phase of oogenesis, ornithine decarboxylase expression is regulated at a translational level. These results are discussed relative to the temporal expression of ornithine decarboxylase and of other proteins whose expression also decreases during oogenesis. In order to perform these experiments, the cDNA (XLODC1) corresponding to Xenopus laevis ornithine decarboxylase mRNA was cloned and sequenced.


Modulation of amygdala response to task-irrelevant emotion.
It has been shown that as cognitive demands of a non-emotional task increase, amygdala response to task-irrelevant emotional stimuli is reduced. However, it remains unclear whether effects are due to altered task demands, or altered perceptual input associated with task demands. Here, we present fMRI data from 20 adult males during a novel cognitive conflict task in which the requirement to scan emotional information was necessary for task performance and held constant across levels of cognitive conflict. Response to fearful facial expressions was attenuated under high (vs low) conflict conditions, as indexed by both slower reaction times and reduced right amygdala response. Psychophysiological interaction analysis showed that increased amygdala response to fear in the low conflict condition was accompanied by increased functional coupling with middle frontal gyrus, a prefrontal region previously associated with emotion regulation during cognitive task performance. These data suggest that amygdala response to emotion is modulated as a function of task demands, even when perceptual inputs are closely matched across load conditions. PPI data also show that, in particular emotional contexts, increased functional coupling of amygdala with prefrontal cortex can paradoxically occur when executive demands are lower.


Incubation of trypanosome-derived mitogenic and immunosuppressive products with peritoneal macrophages allows recovery of biological activities from soluble parasite fractions.
This report describes further attempts to define the nature of the parasite product(s) responsible for the extensive changes in lymphoid tissue in mice during infection with Trypanosoma brucei. As previously described, potent mitogenic and immunosuppressive effects are induced by a trypanosome-derived crude membrane fraction in vivo. There was no enrichment in these activities when purified parasite surface membranes were used. Mitogenic activity can be recovered from soluble trypanosome material only when it is incubated with peritoneal macrophages before transfer into syngeneic recipients. Thus, by encouraging association with a critical target cell, soluble parasite products can be studied, and their active components can be separated by conventional methods. Preliminary fractionation of high-spin trypanosome supernatant over Sepharose 4B confined the mitogenic activity to the high-molecular-weight fraction, which is a macromolecular complex of proteins, glycoproteins, and lipid. Extracted lipid from this material was able to significantly suppress a primary immunoglobulin G anti-sheep erythrocyte response. The activity was periodate sensitive and pronase resistant. The use of macrophages in vitro may be a general method whereby important biological activities lost as a result of fractionation procedures can be recovered and the active components studied in greater detail.


Cardiovascular safety of growth hormone treatment in Noonan syndrome: real-world evidence.
The study aims to assess the cardiovascular safety of growth hormone (GH) treatment in patients with Noonan syndrome (NS) in clinical practice. The study design involves two observational, multicentre studies (NordiNet® IOS and the ANSWER Program) evaluating the long-term effectiveness and safety of GH in >38,000 paediatric patients, of which 421 had NS. Serious adverse events, serious adverse reactions (SARs) and non-serious adverse reactions (NSARs) were reported by the treating physicians. Cardiovascular comorbidities at baseline and throughout the studies were also recorded. The safety analysis set comprised 412 children with NS (29.1% females), with a mean (s.d.) baseline age of 9.29 (3.88) years, treated with an average GH dose of 0.047 (0.014) mg/kg/day during childhood. Cardiovascular comorbidities at baseline were reported in 48 (11.7%), most commonly pulmonary valve stenosis (PVS) and atrial septal defects. Overall, 22 (5.3%) patients experienced 34 safety events. The most common were the NSARs: headache (eight events in seven patients) and arthralgia (five events in three patients). Two SARs occurred in one patient (brain neoplasm and metastases to spine). No cardiovascular safety events were recorded in patients with NS. Five cardiovascular comorbidities in five patients were reported after initiation of GH treatment: three cases of unspecified cardiovascular disease, one ruptured abdominal aortic aneurysm and one PVS. GH treatment had a favourable safety profile in patients with NS, including those with cardiovascular comorbidities. Prospective studies are warranted to systematically assess the safety of GH treatment in patients with NS and cardiovascular disease.


[The wonderful history of neonatal screening].
The concept of neonatal screening for metabolic disorders was launched by Robert Guthrie in 1963. A filter paper blood sample and a cheap and rapid analytical technique were factors that paved the road to success. Mass screening for phenylketonuria soon started to spread. Various additional screening procedures for other metabolic or endocrine conditions were subsequently developed. Most of developed countries have set up systematic neonatal screening programs and million of babies are screened each year. This review presents the main milestones of the fantastic history of neonatal screening but also the questions risen by many temptations to extend the screening to other disorders which not necessarily fulfil classic criteria for systematic search. Ethical and health economical aspects will be the most important issues for the development of neonatal screening in the next millennium.


A blueprint for interprofessional learning.
Interprofessional education (IPE) has been promoted as a method to enhance the ability of health professionals to learn to work together. This article examines several approaches to learning that can help IPE fulfill its expectations. The first is aimed at the transfer of learning novel situations and involves two ideas. Students need to be challenged with progressively more complex tasks and those tasks need to reflect the reality in which they will be working. Second, the learning situation needs to be structured using the five elements of best-practice cooperative learning: positive interdependence, face-to-face promotive interaction, individual accountability, interpersonal and small-group skills, and group processing. Finally, the learning process itself needs to be approached from an experiential learning framework cycling through the four-stage model of planning, doing, observing and reflecting. By using increasingly complex and relevant cases in cooperative groups with an experiential learning process interprofessional education can be successful.


[Immune monitoring of kidney transplant recipients: can markers predictive of over-immunosuppression be identified?].
While the use of nonspecific immunosuppressive drugs has significantly reduced the incidence of acute graft rejection, the benefits of such therapies on chronic rejection and overall long-term graft survival are uncertain. Persistent excessive immunosuppression after immunosuppressive drug treatment is associated with long-term toxicity including increased incidence of cancers, severe infectious complications and metabolic diseases (for example, diabetes, atherosclerosis). One of our team's aims is to identify immunological factors that can predict such toxicities. We have previously demonstrated that CD4T cell cytopenia was correlated with high risk of cancers and infections as well as atherosclerosis in renal transplant recipients. Now, we are investigating the mechanisms involved in CD4T cell cytopenia. We are also exploring how inflammation and cells from the innate immunity influence the complications associated with kidney transplantation. This was performed through the analysis of gene polymorphism on TLR-4, NOD2/CARD15 receptors and IL-6 promoter and correlation with transplantation outcome. We already correlated IL-6 promoter gene polymorphism at position -174 with new-onset diabetes after transplantation in overweight patients. Identification of gene polymorphisms or factors associated with complications after transplantation may help physicians to determine high-risk recipient profiles and optimize pre- and post-transplantation treatment strategies.


Pharmacokinetic/pharmacodynamic assessments of 10 mg/kg tramadol intramuscular injection in yellow-bellied slider turtles (Trachemys scripta scripta).
In reptiles, administration of opioid drugs has yielded unexpected results with respect to analgesia. The aims of this study were to assess the pharmacokinetic/pharmacodynamic (PK/PD) properties of tramadol and its active metabolite M1 and to evaluate the effect of the renal portal system on the PK/PD parameters in yellow-bellied slider turtles. Turtles (n = 19) were randomly assigned to four treatment groups, according to a masked, single-dose, four-treatment, unpaired, four-period crossover design. Group A (n = 5) received a single i.m. dose of tramadol (50 mg/mL) at 10 mg/kg in the proximal hindlimb. Group B (n = 5) received the same i.m. dose but in the forelimb. Groups C (n = 5) and D (n = 4) received a single i.m. injection of saline (NaCl 0.9%) of equivalent volume to the volumes of tramadol injected in the hind- and forelimb, respectively. Groups were rotated (1-month washout period) until the completion of the crossover study. Tramadol plasma concentrations were evaluated by a validated HPLC-FL method. An infrared thermal stimulus was applied to the plantar surface of the turtles' hindlimbs to evaluate the thermal withdrawal latency (TWL). The two PK profiles of tramadol differed in the first 2 h following administration, but overlapped in the elimination phases. The metabolite M1 was formed in both the treatment groups, showing similar pharmacokinetic trends, although the amount of M1 was significantly higher (20%) in the hindlimb vs. forelimb group. Turtles given tramadol in the hind- and forelimb showed a significant increase in TWL over the periods of 0.5-48 and 8-48 h, respectively. The calculated % maximal possible response (% MPR) was low (about 24%). The PK/PD correlations between M1 plasma concentrations vs. % MPR appeared to show a counterclockwise hysteresis loop shape.


Antibiotic susceptibility testing of grown blood cultures by combining culture and real-time polymerase chain reaction is rapid and effective.
Early administration of appropriate antibiotic therapy in bacteraemia patients dramatically reduces mortality. A new method for RApid Molecular Antibiotic Susceptibility Testing (RAMAST) that can be applied directly to positive blood cultures was developed and evaluated. Growth curves and antibiotic susceptibility of blood culture isolates (Staphylococcus aureus, enterococci and (facultative) aerobic gram-negative rods) were determined by incubating diluted blood cultures with and without antibiotics, followed by a quantitative universal 16S PCR to detect the presence or absence of growth. Testing 114 positive blood cultures, RAMAST showed an agreement with microbroth dilution of 96.7% for gram-negative rods, with a minor error (false-susceptibility with a intermediate resistant strain) rate of 1.9%, a major error (false resistance) rate of 0.8% and a very major error (false susceptibility) rate of 0.6%. Agreement for S. aureus was 97.9%, with a very major error rate of 2.1%. Enterococcus species showed 95.0% agreement, with a major error rate of 5.0%. These agreements are comparable with those of the Phoenix system. Starting from a positive blood culture, the test was completed within 9 hours. This new rapid method for antibiotic susceptibility testing can potentially provide accurate results for most relevant bacteria commonly isolated from positive blood cultures in less time than routine methods.


Dip Pen Nanolithography (DPN): process and instrument performance with NanoInk's NSCRIPTOR system.
Precision nanoscale deposition is a fundamental requirement for much of current nanoscience research and promises to facilitate exciting industrial applications. Tailoring chemical composition and surface structure on the sub-100 nm scale benefits researchers in topics ranging from catalysis, to biological recognition in nanoscale systems, to electronic connectivity on the nanoscale. Precision nanoscale deposition engenders applications such as additive photomask repair and nanodevice fabrication. Dip Pen Nanolithography (DPN) is a scanning-probe-based direct-write technique for generating surface-patterned chemical functionality and discrete structures on the sub-100 nm scale. In this publication we explore the effects of changing tip radius and surface roughness. We find that blunter tips lead to higher minimum line widths and that higher rms surface roughness leads to higher minimum line widths; line edge roughness also increases with substrate roughness and surface feature size. Also, we characterize the performance of the Nscriptor DPN instrument and demonstrate the placement of pattern features with precision better than 10 nm, and size control better than 15% for sub-100 nm features.


Host Controls of Within-Host Disease Dynamics: Insight from an Invertebrate System.
AbstractWithin-host processes (representing the entry, establishment, growth, and development of a parasite inside its host) may play a key role in parasite transmission but remain challenging to observe and quantify. We develop a general model for measuring host defenses and within-host disease dynamics. Our stochastic model breaks the infection process down into the stages of parasite exposure, entry, and establishment and provides associated probabilities for a host's ability to resist infections with barriers and clear internal infections. We tested our model on Daphnia dentifera and the parasitic fungus Metschnikowia bicuspidata and found that when faced with identical levels of parasite exposure, Daphnia patent (transmitting) infections depended on the strength of internal clearance. Applying a Gillespie algorithm to the model-estimated probabilities allowed us to visualize within-host dynamics, within which signatures of host defense could be clearly observed. We also found that early within-host stages were the most vulnerable to internal clearance, suggesting that hosts have a limited window during which recovery can occur. Our study demonstrates how pairing longitudinal infection data with a simple model can reveal new insight into within-host dynamics and mechanisms of host defense. Our model and methodological approach may be a powerful tool for exploring these properties in understudied host-parasite interactions.


Bordetella adenylate cyclase: host toxicity and diagnostic utility.
Two toxins from Bordetella pertussis, pertussis toxin and Bordetella adenylate cyclase, cause profound disruptions of cAMP metabolism in mammalian cells. While the role of each toxin in the whooping cough syndrome is unknown, it is highly likely that together they confer on the organism an important proliferative advantage. Intact B. pertussis cells express large amounts of adenylate cyclase activity on their exterior surface. In a presently unknown fashion, this enzyme can enter human phagocytes, elevate cellular cAMP and impair host defense. We reasoned that this unusual enzyme might serve to signal the presence of B. pertussis in nasopharyngeal swabs from infected persons. Here we report a series of in vitro experiments which confirm the feasibility of such an approach. We find that calcium alginate swabs containing as few as 100 B. pertussis organisms produce readily detectable amounts of cAMP in our assay. Nasopharyngeal secretions swabbed from healthy volunteers, however produced no detectable cAMP alone and did not affect the production of cAMP by B. pertussis. We have also tested pure cultures of four common bacterial pathogens (Haemophilus influenzae, Streptococcus pyogenes, Staphylococcus aureus, and Escherichia coli) which may be co-isolated in clinical whooping cough and find that none interferes with the detection of B. pertussis. We conclude that the unique adenylate cyclase of B. pertussis might be a valuable diagnostic device.


Optogenetic LED array for perturbing cardiac electrophysiology.
Optogenetics is the targeted genetic introduction of light-sensitive channels, such as Channelrhodopsin, and pumps, such as Halorhodopsin, into electrically-excitable cells that enables high spatiotemporal electrical stimulation and inhibition by optical actuation. Technologies for inducing optogenetically-based electrical stimulation for investigating in vitro and in vivo neural perturbations have been described. However, modification of existing technologies or creation of new ones has not been described for chronic cardiac applications. Here, an LED array system for optogenetically perturbing cardiac electrophysiology is described. The overall layout of the system consists of an LED holder containing six LED's that deliver pulsed ∼470 nm light to pluripotent stem cell-derived cardiomyocytes cultured in a 6-well tissue culture plate. The response of the cardiomyocytes is monitored by microscopy and the system is enclosed within a standard incubator. This system is relatively simple to create and uses mostly off-the-shelf components. The overall function of the system is to deliver chronic light stimulation over days to weeks to differentiating stem cell-derived cardiomyocytes in order to investigate perturbations in their electrophysiology.


Effect of Curcumin Supplementation on Exercise-Induced Oxidative Stress, Inflammation, Muscle Damage, and Muscle Soreness.
Curcumin has been shown to reduce exercise-induced oxidative stress (OS) and inflammation. The purpose of this investigation was to examine the effects of curcumin supplementation on OS, inflammation, muscle damage, and muscle soreness. Nineteen males participated in a randomized, double-blinded, placebo-controlled trial to examine the effects of curcumin supplementation (1.5 g/day) compared to a placebo (PLA) following a muscle-damaging protocol (MDP) on OS, inflammation, muscle damage, and soreness. Participants were randomized to two groups, curcumin or placebo group. The MDP was performed before and after supplementation (28 days). Blood was sampled pre- and postexercise and 60 min, 24 h, and 48 h postexercise and analyzed for total antioxidant capacity (TAC), malondialdehyde (MDA), tumor necrosis factor alpha (TNF-α), and creatine kinase (CK). In addition, a visual analog scale (VAS) was used on each blood sample to measure perceived muscle soreness. After supplementation, curcumin significantly blunted CK levels (199.62 U/L) compared to the placebo (287.03 U/L), overall (p < 0.0001). In addition, curcumin resulted in decreased muscle soreness, overall (VAS scale 2.88), when compared to the placebo (VAS scale 3.36) (p = 0.0120). There were no differences found in TAC, TNF-α, or MDA. Curcumin may reduce muscle damage and perceived muscle soreness without negatively impacting a natural inflammatory response following exercise. Future research should investigate chronic curcumin supplementation and its mechanistic effects on muscle recovery from exercise.


[Transumbilical cholecystectomy].
Recent development of minimally invasive surgery has raised interest in laparoscopic surgery through a single abdominal incision. The most common names for this technique is single incisional laparoscopic surgery (SILS) or laparoscopic endoscopic single-site surgery (LESS). This article describes the first experiences at our department with cholecystectomy using this new technique. Twelve patients underwent cholecystectomy with SILS technique at Bispebjerg Hospital. The exclusion criteria were previous abdominal surgery and known previous cholecystitis. The median operation time was 89 minutes. Four patients presented signs of cholecystitis or adhesions. There was no perioperatives complications. Ten patients were discharged at the day of the operation and two the day after. This patient series reveals that SILS technique is feasible in selected patients. It yields an optimal cosmetic result, but conclusions about the potential benefits regarding postoperative pain, nausea, port-site bleeding, infection, port-site hernia and bile duct lesions as compared with conventional laparoscopic cholecystectomy remain unclear. Randomized trials are warranted to answer these questions.


Changes in lectin-binding patterns during late fetal development of the rat colon.
Lectin-binding and histochemical studies were integrated with a morphological description of colon development in rat fetuses to determine whether changes in glycoprotein sugars could be identified with stages of colon organogenesis. At 16 days gestation the colon consisted of a minute lumen surrounded by 3-5 layers of cuboidal, stratified epithelium, a basement membrane and undifferentiated mesenchyme. As development proceeded, epithelial spaces eventually fused with the main lumen, and fingers of mesenchyme, with a basement membrane and epithelial mantle, formed the walls of crypts lined with simple to stratified columnar cells. Goblet cells and mucin production appeared only on the 20th-21st day of gestation. Mesenchyme differentiation to a circular muscle band, the prospective tunica muscularis, occurred on days 17-18, and vascularization of the lamina propria was first detected on day 19. Wheat germ agglutinin (WGA) lectin bound avidly to sugar residues in the mesenchyme matrix from day 16 but weakened as the tissue differentiated, particularly into smooth muscle. Alcian blue staining of the matrix, also weakening with time, confirmed the sialic acid nature of binding sites for WGA on matrix acid mucopolysaccharides. Under controlled conditions, WGA also detected N-acetylglucosamine-binding sites on brush borders as they developed on the apical surface of primitive enterocytes. Ulex lectin bound to fucose residues of Alcian-blue-positive, PAS-positive mucin from the first appearance of goblet cells. Concanavalin A and glycine max lectins bound only to blood group substances on both nucleated red cells between days 16 and 19 and anucleate red cells from day 19.(ABSTRACT TRUNCATED AT 250 WORDS)


A transgenic mouse model with an inducible skin blistering disease phenotype.
One of the current limitations of gene transfer protocols involving mammalian genomes is the lack of spatial and temporal control over the desired gene manipulation. Starting from a human keratin gene showing a complex regulation as a template, we identified regulatory sequences that confer inducible gene expression in a subpopulation of keratinocytes in stratified epithelia of adult transgenic mice. We used this cassette to produce transgenic mice with an inducible skin blistering phenotype mimicking a form of epidermolytic hyperkeratosis, a keratin gene disorder. Upon induction by topical application of a phorbol ester, the mutant keratin transgene product accumulates in the differentiating layers of epidermis, leading to keratinocyte lysis after application of mechanical trauma. This mouse model will allow for a better understanding of the complex relationship between keratin mutation, keratinocyte cytoarchitecture, and hypersensitivity to trauma. The development of an inducible expression vector showing an exquisite cellular specificity has important implications for manipulating genes in a spatially and temporally controlled fashion in transgenic mice, and for the design of gene therapy strategies using skin as a tissue source for the controlled delivery of foreign substances.


Hypotension 1.5 min after 10% hemorrhage permits evaluation of rabbit's baroreflex.
In seven chronically instrumented rabbits, we studied the influence of the vasopressin system on the evaluation of acute reflex compensation of arterial pressure for a mild hemorrhage. Rabbits were bled 10% of estimated blood volume in 1 min. The hemorrhage was repeated before and after administration of a vasopressin antagonist [d(CH2)5Tyr(Me)arginine vasopressin, 10 micrograms/kg] and in the conscious and anesthetized (pentobarbital sodium 20 mg/kg) state. The efficacy of the vasopressin antagonist was preliminarily tested in each rabbit by confirming its antagonism against the hemodynamic effects of infused vasopressin (50 ng X min-1 X kg-1). Mean arterial pressure (MAP), heart rate (HR), mean aortic flow (MAF), and total peripheral resistance (TPR) were measured before and at 1.5 min after completion of hemorrhage. The hemorrhage caused statistically significant decrease in MAP (84 +/- 6 to 78 +/- 6 mmHg) and MAF (195 +/- 28 to 158 +/- 26 ml X kg-1 X min-1) and increase in TRP (0.44 +/- 0.05 to 0.51 +/- 0.07 mmHg X kg X min X ml-1) and HR (271 +/- 21 to 284 +/- 22 beat/min). However, there was no statistically significant interaction in any variables between vasopressin and hemorrhage, i.e., whether vasopressin was blocked or intact did not affect the fall in MAP by hemorrhage. We conclude that in the rabbit vasopressin's role in the acute compensation of arterial pressure for a quick 10% hemorrhage is small enough to permit accurate estimation of the arterial baroreflex gain from the depressor responses to mild hemorrhage with and without the reflex function.


Requirement of open headpiece conformation for activation of leukocyte integrin alphaXbeta2.
Negative stain electron microscopy (EM) and adhesion assays show that alpha(X)beta(2) integrin activation requires headpiece opening as well as extension. An extension-inducing Fab to the beta(2) leg, in combination with representative activating and inhibitory Fabs, were examined for effect on the equilibrium between the open and closed headpiece conformations. The two activating Fabs stabilized the open headpiece conformation. Conversely, two different inhibitory Fabs stabilized the closed headpiece conformation. Adhesion assays revealed that alpha(X)beta(2) in the extended-open headpiece conformation had high affinity for ligand, whereas both the bent conformation and the extended-closed headpiece conformation represented the low affinity state. Intermediate integrin affinity appears to result not from a single conformational state, but from a mixture of equilibrating conformational states.


Clustering in image space for place recognition and visual annotations for human-robot interaction.
The most classical way of attempting to solve the vision-guided navigation problem for autonomous robots corresponds to the use of three-dimensional (3-D) geometrical descriptions of the scene; what is known as model-based approaches. However, these approaches do not facilitate the user's task because they require that geometrically precise models of the 3-D environment be given by the user. In this paper, we propose the use of "annotations" posted on some type of blackboard or "descriptive" map to facilitate this user-robot interaction. We show that, by using this technique, user commands can be as simple as "go to label 5." To build such a mechanism, new approaches for vision-guided mobile robot navigation have to be found. We show that this can be achieved by using mixture models within an appearance-based paradigm. Mixture models are more useful in practice than other pattern recognition methods such as principal component analysis (PCA) or Fisher discriminant analysis (FDA)-also known as linear discriminant analysis (LDA), because they can represent nonlinear subspaces. However, given the fact that mixture models are usually learned using the expectation-maximization (EM) algorithm which is a gradient ascent technique, the system cannot always converge to a desired final solution, due to the local maxima problem. To resolve this, a genetic version of the EM algorithm is used. We then show the capabilities of this latest approach on a navigation task that uses the above described "annotations."


Impact of the statin escape phenomenon on long-term clinical outcomes in patients with acute myocardial infarction: Subgroup analysis of the Nagoya Acute Myocardial Infarction Study (NAMIS).
Statins are reportedly effective in the primary and secondary prevention of cardiovascular disease, mainly due to their ability to aggressively reduce low-density lipoprotein cholesterol (LDL-C) levels. However, patients sometimes exhibit the so-called "statin escape" phenomenon. The purpose of our study was to investigate the impact of the statin escape phenomenon on long-term clinical outcomes in patients with acute myocardial infarction (AMI). This was a subgroup analysis of 1144 patients from the Nagoya Acute Myocardial Infarction Study (NAMIS) treated between January 2004 and December 2012. We analyzed 660 patients who initiated statin treatment after AMI. Statin escape phenomenon was defined as an increase in the LDL-C levels during the 9-month treatment period by >10% of the initial values after 4 weeks of initiating statin treatment. Patients were divided into two groups depending on whether they exhibited the statin escape phenomenon, with 474 patients in the non-escape group and 186 patients in the escape group. Compared to the non-escape group, the escape group showed significantly lower LDL-C levels at 4 weeks after treatment initiation (81.3 ± 20.1 mg/dL vs. 101.1 ± 25.4 mg/dL, P < 0.01). By contrast, the escape group showed significantly higher LDL-C levels at 9 months after treatment initiation (105.8 ± 28.3 mg/dL vs. 90.3 ± 22.6 mg/dL, P < 0.01). Major adverse cardiac and cerebrovascular events (MACCE; a composite of all-cause death, MI, and stroke) were more frequent in the escape group than in the non-escape group (10.8% vs. 6.1%, P = 0.03). Multivariate analysis showed that statin escape phenomenon was an independent predictor of MACCE (hazard ratio: 2.02, 95% confidence interval: 1.11-3.66, P = 0.02). Statin escape phenomenon may be an independent predictor of long-term clinical outcomes in AMI patients.


It is diprotonated inorganic phosphate that depresses force in skinned skeletal muscle fibers.
The increases in the intracellular concentrations of inorganic phosphate and hydrogen ion accompanying fatigue of skeletal muscle appear to be the most important metabolic changes associated with the decrease in contractile force. Experiments on chemically skinned single fibers from rabbit psoas muscle with pH ranging between 6 and 7.25 demonstrate that the depression of maximal calcium-activated force by inorganic phosphate correlates nicely with the concentration of the acidic (diprotonated) species. Therefore, in addition to the well-known depressant effect on the contractile machinery of lowering pH per se, any decrease of intracellular pH associated with fatigue further depresses force production by converting more of the total inorganic phosphate within the cell to the inhibitory diprotonated form.


Tracking discussions of complementary, alternative, and integrative medicine in the context of the COVID-19 pandemic: a month-by-month sentiment analysis of Twitter data.
Coronavirus disease 2019 (COVID-19) is a novel infectious disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Despite the paucity of evidence, various complementary, alternative and integrative medicines (CAIMs) have been being touted as both preventative and curative. We conducted sentiment and emotion analysis with the intent of understanding CAIM content related to COVID-19 being generated on Twitter across 9 months. Tweets relating to CAIM and COVID-19 were extracted from the George Washington University Libraries Dataverse Coronavirus tweets dataset from March 03 to November 30, 2020. We trained and tested a machine learning classifier using a large, pre-labelled Twitter dataset, which was applied to predict the sentiment of each CAIM-related tweet, and we used a natural language processing package to identify the emotions based on the words contained in the tweets. Our dataset included 28 713 English-language Tweets. The number of CAIM-related tweets during the study period peaked in May 2020, then dropped off sharply over the subsequent three months; the fewest CAIM-related tweets were collected during August 2020 and remained low for the remainder of the collection period. Most tweets (n = 15 612, 54%) were classified as positive, 31% were neutral (n = 8803) and 15% were classified as negative (n = 4298). The most frequent emotions expressed across tweets were trust, followed by fear, while surprise and disgust were the least frequent. Though volume of tweets decreased over the 9 months of the study, the expressed sentiments and emotions remained constant. The results of this sentiment analysis enabled us to establish key CAIMs being discussed at the intersection of COVID-19 across a 9-month period on Twitter. Overall, the majority of our subset of tweets were positive, as were the emotions associated with the words found within them. This may be interpreted as public support for CAIM, however, further qualitative investigation is warranted. Such future directions may be used to combat misinformation and improve public health strategies surrounding the use of social media information.


Study of cell wall growth in Bacillus megaterium by high-resolution autoradiography.
Growth of the cell wall of Bacillus megaterium was studied by pulse-labeling the cell wall of a DAP- Lys- mutant for a very short time with tritium-labeled diaminopimelic acid. The distribution of radioactivity along the cell wall was examined by high-resolution autoradiography on isolated cell walls and thin sections of bacteria. The results indicate that cell wall elongation occurs by diffuse intercalation of newly synthesized murein into the expanding cell wall during exponential growth, as well as during germination, and that the only zone of highly localized diaminopimelic acid incorporation is found at the cross wall during its synthesis. This zone contains about 30% of the radioactivity incorporated into the cell wall. Analysis of autoradiographs of thin sections of bacteria shows that the total radioactivity incorporated per bacterium doubles during the life cycle. This doubling occurs in the cylindrical part of the cell wall but not in the polar caps. This seems to indicate that elongation of the bacterium is not constant during the life cycle but increases with the length of the cell.


Tessellation of porphyrazines with porphyrins by design.
The efficient self-assembly and functional characterization of arrays containing multiple types of chromophores will provide a basis for the design and applications of functional photonic materials that are unobtainable using only one type of molecule. The design, synthesis, and characterization of supramolecular systems bearing two different types of porphyrinic chromophores, porphyrins and porphyrazines, are reported. Because the porphyrins and porphyrazines bear different exocyclic ligands for self-assembly by metal ion coordination, these systems require new supramolecular synthetic strategies wherein reactants are added in a specific order. These arrays display unique photophysical properties derived from the component chromophores, the metal geometry, and the supramolecular nanoarchitecture.


Molecular characterization of mycobacteria isolated from seals.
Tuberculosis (TB) was diagnosed in 10 seals from three species (Arctocephalus australis, Arctocephalus tropicalis and Otaria flavescens) found in South America. The mycobacteria isolated from these cases belonged to the Mycobacterium tuberculosis complex, as determined by RFLP using an IS6110 probe, spoligotyping, analysis of the 16S rRNA gene sequence and by PCR-restriction analysis of hsp65. Polymorphisms in gyrA, katG, oxyR and pncA were investigated in some of the isolates, as well as the presence of the MPB70 antigen. The insertion sequence IS6110 was present in three to seven copies in the genome of the mycobacteria isolated from seals. Using the IS6110 probe, six patterns (designated A, B, C, D, E and F) were identified from 10 different isolates. Patterns A and B were found for the mycobacteria isolated from two and four seals, respectively, indicating an epidemiological relationship between isolates grouped according to their IS6110 RFLP. The mycobacteria isolated from seals shared the majority of their IS6110 DNA-containing restriction fragments, and nine isolates had an identical spoligotype; only one isolate showed a minor difference in its spoligotype. In addition, none of these spoligotypes were found in other M. tuberculosis complex strains. These results suggest that the isolates from seals constitute a unique group of closely related strains. The mycobacteria isolated from seals showed polymorphisms at gyrA codon 95 and katG codon 463, as do group 1 M. tuberculosis, and M. bovis. Group 1 mycobacteria are associated with cluster cases. The spoligotypes found in the mycobacteria isolated from seals lack spacers 39-43, as does M. bovis, but the MPB70 antigen, which is highly expressed in M. bovis and minimally expressed in M. tuberculosis, was not detected in these mycobacteria. The mycobacteria isolated from seals also showed oxyR and pncA polymorphisms specific to M. tuberculosis. In conclusion, the mycobacteria that cause TB in seals in the South-Western Atlantic are a related group, and based on the combination of genetic characteristics, belong to a unique genotypic group within the M. tuberculosis complex.


Decreased working memory and processing speed mediate cognitive impairment in geriatric depression.
While neuropsychological dysfunction is common in geriatric depression, not all aspects of cognition are equally affected. It has been suggested that depressed patients are impaired only in tasks that make heavy demands on processing resources and that a resource decrement therefore underlies the neuropsychological decrements seen in geriatric depression. The present study examined whether processing resources in the form of working memory and information processing speed are decreased in depression and whether a decrease in these resources actually mediates neuropsychological impairment. Measures of processing resources were administered to elderly depressed patients prior to treatment and to age-matched controls. Patients whose depression remitted were retested as were the controls. Subjects also received neuropsychological tests of episodic memory and visuospatial performance. Depressed patients performed significantly worse on measures of both processing speed and working memory. While performance on these measures improved in patients whose depression remitted, the amount of improvement was no greater than that seen in the controls with repeat testing. Hierarchical regression analyses showed that depression explained a significant amount of variance on the neuropsychological tasks. However, if the variance associated with processing resources was removed first, depression no longer accounted for a significant amount of neuropsychological variance. Processing resources are decreased in elderly depressed patients and this decrease in resources appears to mediate impairments in several areas of neuropsychological functioning including episodic memory and visuospatial performance. The resource decrement persists after remission of the depression and thus may be a trait marker of geriatric depression.


[Guidelines for the management of painful endometriosis].
To establish guidelines for the medical and surgical management of painful endometriosis. An exhaustive review on Medline and Cochrane Database between 1980 and 2006 was performed. GnRH agonists, progestins, continuous monophasic oral contraceptives and danazol have a suppressive effect on dysmenorrhoea, nonmenstrual pain and dyspareunia (grade A). Surgical treatment is effective in painful endometriosis (grade B). Complete surgical excision of deep endometriotic lesions with conservation of uterus and ovaries has a limited term efficacy on pain relief (grade C). A multidisciplinary approach is recommended (grade C). The use of the psychotherapy improves the management of chronic pain (grade A). There is a lack of information concerning the therapeutic strategy able to prevent recurrences. Whether endometriosis recurrences occur, medical treatment should be the first line approach (expert opinion). A hysterectomy with salpingo-oophorectomy and complete excision of the lesions is efficient in women with pain recurrence who no longer desire pregnancy (grade C). Medical and surgical treatments have a limited term efficacy on painful endometriosis (grade A). The benefit/risk ratio, depending on side-effects, should be assessed on a case to case basis.


The efficacy of intra-articular hyaluronan injection after the microfracture technique for the treatment of articular cartilage lesions.
Although the exact mechanism of action has yet to be elucidated, recent animal studies have demonstrated chondroprotective and anti-inflammatory properties of hyaluronic acid viscosupplementation. Intra-articular hyaluronic acid after microfracture improves the quality of the repair leading to a more hyaline-like repair tissue with better defect fill and adjacent area integration. Controlled laboratory study. Full-thickness cartilage defects were created in the weightbearing area of the medial femoral condyle in 36 female New Zealand White rabbits. The defects were then treated with surgical microfracture. Eighteen rabbits formed the 3-month cohort and the other 18 formed the 6-month cohort. Within each cohort, 6 rabbits were randomly assigned to receive 3 weekly injections of hyaluronic acid (group A), 5 weekly injections (group B), or control injections of normal saline (group C). At 3 and 6 months postmicrofracture, the animals were sacrificed and the operative knee harvested. Repair tissue was assessed blinded- both grossly, using a modified component of the International Cartilage Repair Society (ICRS) Cartilage Repair Assessment scoring scale, and histologically, using the modified O'Driscoll histological cartilage scoring system. Comparisons were made with respect to gross and histologic findings between treatment groups at each time point. Effects of each treatment type were also evaluated longitudinally by comparing the 3-month results with the 6-month results. Statistical analysis was performed using unpaired Student t tests with significance defined as P < .05. At 3 months, gross and histologic evaluation of the repair tissue demonstrated that the 3-injection group had significantly better fill of the defects and more normal appearing, hyaline-like tissue than controls (a mean ICRS score of 1.92 vs 1.26; P < .05 and a mean modified O'Driscoll score of 10.3 vs 7.6; P < .02). Specimens treated with 5 weekly injections were not significantly improved compared with controls. At 6 months, the mean gross appearance and histologic scores between the 3 specimen cohorts were not significantly different. However, examination of the entire operative knee demonstrated a significantly greater extent of degenerative changes (synovial inflammation and osteophyte formation) in the control group than in both hyaluronic acid treatment groups (P < .05). Supplementing the microfracture technique with 3 weekly injections of intra-articular hyaluronic acid had a positive effect on the repair tissue that formed within the chondral defect at the early follow-up time point. This improvement was not found for the 3-injection group at 6 months or for the 5-injection group at either time point. Additionally, hyaluronic acid supplementation had a possible chondroprotective and anti-inflammatory effect, limiting the development of degenerative changes within the knee joint. The adjunctive use of hyaluronic acid appears to hold promise in the treatment of chondral injuries and warrants further investigation.


Methodological quality of systematic reviews referenced in clinical practice guidelines for the treatment of opioid use disorder.
With efforts to combat opioid use disorder, there is an increased interest in clinical practice guidelines (CPGs) for opioid use disorder treatments. No literature exists examining the quality of systematic reviews used in opioid use disorder CPGs. This study aims to describe the methodological quality and reporting clarity of systematic reviews (SRs) used to create CPGs for opioid use disorder. From June to July 2016 guideline clearinghouses and medical literature databases were searched for relevant CPGs used in the treatment of opioid use disorder. Included CPGs must have been recognized by a national organization. SRs from the reference section of each CPG was scored by using AMSTAR (a measurement tool to assess the methodological quality of systematic reviews) tool and PRISMA (preferred reporting items for systematic reviews and meta-analyses) checklist. Seventeen CPGs from 2006-2016 were included in the review. From these, 57 unique SRs were extracted. SRS comprised 0.28% to 17.92% of all references found in the CPGs. All SRs obtained moderate or high methodological quality score on the AMSTAR tool. All reviews met at least 70% of PRISMA criteria. In PRISMA, underperforming areas included accurate title labeling, protocol registration, and risk of bias. Underperforming areas in AMSTAR included conflicts of interest, funding, and publication bias. A positive correlation was found between AMSTAR and PRISMA scores (r = .79). Although the SRs in the CPGs were of good quality, there are still areas for improvement. Systematic reviewers should consult PRISMA and AMSTAR when conducting and reporting reviews. It is important for CPG developers to consider methodological quality as a factor when developing CPG recommendations, recognizing that the quality of systematic reviews underpinning guidelines does not necessarily correspond to the quality of the guideline itself.


NIR fluorescence labelled carbon nano-onions: synthesis, analysis and cellular imaging.
The preparation of novel NIR fluorescent carbon based nanomaterials, consisting of boron difluoride azadipyrromethene fluorophores covalently attached to carbon nano-onions, is demonstrated. In addition, the analysis of the new nanomaterial is presented. The fluorescent nano-derivative properties are customized such that their emission can be reversibly on/off modulated in response to pH, which is demonstrated in solution and in cells. The in vitro imaging of HeLa Kyoto cells is carried out and the cellular uptake of the carbon nano-onion NIR fluorophore conjugates is verified.


The breaking bad effect: Priming with an antihero increases sensation seeking.
Antiheroes are characters that share features with both heroes and villains, typified as selfish and rule breakers, but who end up doing something good for society. In this research, we examined how priming people with antiheroes (vs. heroes) affected their sensation seeking. We reason that antiheroes (vs. heroes) are more associated with temporally close (vs. past and future) events. Given that sensation seeking is related to being focused on the present (vs. past or future), we hypothesized that if people are primed with antiheroes (vs. heroes) they are more likely to seek sensation. Findings from a series of five experimental studies provide insights into the effect of priming with an antihero on people's sensation seeking, providing directions for future research in psychology and practical applications in the areas of marketing strategy and consumer behaviour.


Enhanced gap filling and osteoconduction associated with alendronate-calcium phosphate-coated porous tantalum.
Porous tantalum has been shown to be effective in achieving bone ingrowth. However, in some circumstances, bone quality or quantity may be insufficient to allow adequate bone ingrowth. We hypothesized that local delivery of alendronate from porous tantalum would enhance the ability of the tantalum to achieve bone ingrowth when there is a gap between the implant and bone. We evaluated the effect of alendronate-coated porous tantalum on new bone formation in an animal model incorporating a gap between the implant and bone. A cylindrical porous tantalum implant was implanted in the distal part of each femur in eighteen rabbits (a total of thirty-six implants) and left in situ for four weeks. Three types of porous tantalum implants were inserted: those with no coating (the control group), those with microporous calcium phosphate coating, and those coated with microporous calcium phosphate and alendronate. Subcutaneous fluorescent labeling was used to track new bone formation. Bone formation was analyzed with backscattered electron microscopy and fluorescent microscopy of undecalcified samples. The relative increases in the mean volume of gap filling, bone ingrowth, and total bone formation in the group treated with the porous tantalum implants coated with calcium phosphate and alendronate were 143% (p < 0.001), 259% (p < 0.001), and 193% (p < 0.001), respectively, compared with the values in the control group treated with the uncoated porous tantalum implants. The percentage of the length of the implant that was in contact with new bone in the group treated with the calcium phosphate and alendronate coating was increased by an average of 804% compared with the percentage in the group treated with the uncoated implants. The study demonstrated significantly enhanced filling of the bone-implant gap and bone ingrowth in association with the porous tantalum implants coated with calcium phosphate and alendronate.


Modulation of porcine adipocyte beta-adrenergic receptors by a beta-adrenergic agonist.
Mammalian cells have several mechanisms to decrease the response to beta-adrenergic agonists. Agonists are metabolized or taken up by nerve endings. The beta-adrenergic receptors (betaAR) are inactivated by phosphorylation and removed from the cell membrane, and synthesis is decreased or degradation is increased. Knowledge about adipocyte betaAR desensitization is mostly from rodent adipocytes with > or = 90% beta3AR. Porcine adipocyte betaAR have functional and ligand-binding properties that are quite different from those in many other species. Furthermore, the predominant betaAR subtype in the porcine adipocyte is the beta1AR (70 to 80%). Given these species differences, it might be expected that desensitization in porcine adipocytes would not be totally concordant with the rodent-derived model. Isolated porcine adipocytes were incubated without or with a betaAR agonist, isoproterenol. The total betaAR number, measured by ligand-binding in a crude membrane fraction, tended to be lower after 6 h of incubation without isoproterenol. The addition of 10(-5) M isoproterenol during the incubation caused the betaAR number to decrease 43% compared to cells incubated without isoproterenol. The beta1AR and beta2AR transcript concentrations both decreased 45% after 6 h of incubation without isoproterenol. There was no decrease in mRNA when cells were incubated with isoproterenol. The results suggest the betaAR were desensitized by incubation with isoproterenol, perhaps by phosphorylation and removal from the membrane, but this was not accompanied by modulation of the concentration of transcripts for beta1AR or beta2AR.


Regulation of phagosomal acidification. Differential targeting of Na+/H+ exchangers, Na+/K+-ATPases, and vacuolar-type H+-atpases.
Vacuolar-type (V) ATPases are thought to be the main determinant of phagosomal acidification. In phagosomes containing mycobacteria, which ostensibly impair the delivery of V-ATPases to the phagosomal membrane, the pH would be expected to be near neutral. This prediction was tested by microfluorescence ratio imaging using macrophages from mice susceptible to mycobacterial infection. Although less acidic than their counterparts containing dead bacteria, phagosomes containing live Mycobacteria bovis were nearly 1 pH unit more acidic than the cytosol, suggesting the existence of alternate H+ transport mechanisms. We therefore investigated whether Na+/H+ exchange (NHE) contributes to phagosomal acidification. Immunoblotting, reverse transcriptase-polymerase chain reaction, and pharmacological studies indicated that NHE1 is the predominant isoform of the exchanger in macrophages. Fractionation revealed that NHE1 is incorporated into the phagosomal membrane, and measurements of pH indicated that it is functional in this location. Nevertheless, acidification of the lumen of phagosomes containing either latex beads or live M. bovis was insensitive to (3-methylsulfonyl-4-piperidinobenzoyl)-guanidine methanesulfonate, a potent inhibitor of NHE1. This may have been due to the absence of an appropriate lumen to cytosol Na+ gradient, because the phagosomal membrane was found to be devoid of Na+/K+ pumps. Unexpectedly, the acidification of M. bovis phagosomes was fully reversed by specific inhibitors of the vacuolar H+-ATPase, suggesting that ATPases are present only transiently or in reduced quantities in the phagosomal membrane. Alternatively, acid equivalents accumulated in endosomes by V-ATPases may be delivered to the mycobacterial phagosome by carrier vesicles devoid of ATPases.


Sampling design for the Birth in Brazil: National Survey into Labor and Birth.
This paper describes the sample design for the National Survey into Labor and Birth in Brazil. The hospitals with 500 or more live births in 2007 were stratified into: the five Brazilian regions; state capital or not; and type of governance. They were then selected with probability proportional to the number of live births in 2007. An inverse sampling method was used to select as many days (minimum of 7) as necessary to reach 90 interviews in the hospital. Postnatal women were sampled with equal probability from the set of eligible women, who had entered the hospital in the sampled days. Initial sample weights were computed as the reciprocals of the sample inclusion probabilities and were calibrated to ensure that total estimates of the number of live births from the survey matched the known figures obtained from the Brazilian System of Information on Live Births. For the two telephone follow-up waves (6 and 12 months later), the postnatal woman's response probability was modelled using baseline covariate information in order to adjust the sample weights for nonresponse in each follow-up wave.


Enhancing isoprene polymerization with high activity and adjustable monomer enchainment using cyclooctyl-fused iminopyridine iron precatalysts.
In this study, a series of structurally rigid cyclooctyl-fused iminopyridine iron complexes, [L2FeCl][FeCl4] and [2L3Fe][Cl][3FeCl4], was synthesized via a one-pot method and investigated as precatalysts in conjunction with methylaluminoxane for isoprene (Ip) polymerization. Combined characterization through FTIR analysis, elemental analysis and single crystal XRD analysis fully verified the structure of these complexes. The most active iron complex, FeH, exhibited a trisligated nature, with its cation adopting an octahedral geometry around the metal center. In contrast, all the other iron complexes (Fe2Me, Fe2Et, Fe2iPr, Fe3Me, Fe2Et,Me) displayed bisligated configurations, with distorted trigonal bipyramidal geometry of cations. During isoprene polymerization, the extent of steric hindrance of the ligand framework exerted a significant impact on catalytic performance. The FeH precatalyst with less steric hindrance demonstrated excellent performance, producing high molecular weight polyisoprenes with conversions exceeding 99% for 4000 equiv. of monomer. Even at very low catalyst loadings, as low as 0.0025 mol% (Fe/Ip), the polymerization of isoprene could proceed smoothly with an exceptionally high activity of 4.0 × 106 gPI (molFe, h)-1. Moreover, this precatalyst exhibited good thermal stability, maintaining high activity levels (typically 105 gPI (molFe, h)-1) across a broad temperature range from -20 °C to 100 °C. Additionally, by adjusting steric substituents and the reaction temperature, the 1,4/3,4 regioselectivity could be modulated from 9/91 to 69/31 while maintaining a high stereoselectivity of cis-1,4 structures (cis/trans: >99/1).


CD133+ stem cell mobilization after partial hepatectomy depends on resection extent and underlying disease.
Bone marrow stem cells (BMSC) can participate to liver regeneration. However, conflicting results have been reported on this topic in patients undergoing liver resection. To assess the impact of liver resection extent and presence of underlying liver disease in modulating BMSC mobilization. We enrolled 29 patients undergoing liver resection of different extents, 5 surgical controls and 10 blood donors. Circulating CD133+ BMSC were measured by flow cytometry at different time-points after surgery. The hepatic commitment of mobilized BMSC was investigated by polymerase chain reaction. Liver specimens were collected during surgery for histopathological analysis. Hepatocyte growth factor and granulocyte-colony stimulating factor serum levels were measured by enzyme-linked immunosorbent assay. BMSC mobilization was found in patients undergoing major liver resection, especially in the presence of underlying disease. Ductular reactions were noted in patients with chronic hepatopathy and the hepatic progenitor-like cells expressed CD133, NCAM, cytokeratin-19, and alpha-fetoprotein. Hepatocyte growth factor and granulocyte-colony stimulating factor levels increased following liver resection and the contemporaneous presence of liver disease was associated with their highest raise. Liver repair is mainly an endogenous process. BMSC become important in case of extensive resection, especially in the presence of underlying hepatopathy and hepatic progenitor-like cells activation. Hepatocyte growth factor and granulocyte-colony stimulating factor seem to be involved in the dynamics underlying hepatic regeneration and BMSC recruitment.


Efficient and specific transduction of cochlear supporting cells by adeno-associated virus serotype 5.
Congenital deafness, affecting 1 in 1000 neonates, can lead to major problems in speech, cognitive and psychosocial development. Congenital deafness is mainly caused by mutations in connexins, hemi-channel proteins forming gap-junctions between supporting cells in the sensory epithelia. We describe a high tropism of AAV5 serotype for the supporting cells of the cochlea, both in vitro in postnatal day 4 mouse explants, and in vivo in the adult guinea-pig inner ear, through scala media perfusion. AAV5 transduction correlates with PDGFRalpha expression, previously reported as AAV5 receptor. This vector could be of major interest in addressing gene therapy approaches to deafness as well as for studying basic aspects of inner-ear development and hearing mechanisms.


Sexuality and cancer. Breaking the silence.
Family physicians can be instrumental in helping their patients adjust to changes in sexuality subsequent to treatment for cancer, only if they break the silence surrounding sexuality. Cancer, like other illnesses, can interfere with intimate relationships. Many patients want to discuss changes in their sexuality with their family physician. However, few will raise the issue themselves but wait for their physician to broach the subject. This article summarises the P-LI-SS-IT Model which provides practical guidance on assessing the sexual concerns of cancer patients and on developing helpful interventions.


Glucocorticoids with low-dose anti-IL1 anakinra rescue in severe non-ICU COVID-19 infection: A cohort study.
The optimal treatment for patients with severe coronavirus-19 disease (COVID-19) and hyper-inflammation remains debated. A cohort study was designed to evaluate whether a therapeutic algorithm using steroids with or without interleukin-1 antagonist (anakinra) could prevent death/invasive ventilation. Patients with a ≥5-day evolution since symptoms onset, with hyper-inflammation (CRP≥50mg/L), requiring 3-5 L/min oxygen, received methylprednisolone alone. Patients needing ≥6 L/min received methylprednisolone + subcutaneous anakinra daily either frontline or in case clinical deterioration upon corticosteroids alone. Death rate and death or intensive care unit (ICU) invasive ventilation rate at Day 15, with Odds Ratio (OR) and 95% CIs, were determined according to logistic regression and propensity scores. A Bayesian analysis estimated the treatment effects. Of 108 consecutive patients, 70 patients received glucocorticoids alone. The control group comprised 63 patients receiving standard of care. In the corticosteroid±stanakinra group (n = 108), death rate was 20.4%, versus 30.2% in the controls, indicating a 30% relative decrease in death risk and a number of 10 patients to treat to avoid a death (p = 0.15). Using propensity scores a per-protocol analysis showed an OR for COVID-19-related death of 0.9 (95%CI [0.80-1.01], p = 0.067). On Bayesian analysis, the posterior probability of any mortality benefit with corticosteroids+/-anakinra was 87.5%, with a 7.8% probability of treatment-related harm. Pre-existing diabetes exacerbation occurred in 29 of 108 patients (26.9%). In COVID-19 non-ICU inpatients at the cytokine release phase, corticosteroids with or without anakinra were associated with a 30% decrease of death risk on Day 15.


Absorption kinetics and steady-state plasma concentrations of theophylline following therapeutic doses of two sustained-release preparations.
Ten healthy volunteers received two sustained-release preparations as a single and multiple dose regimen in an open crossover study. Plasma theophylline concentrations were measured by an enzyme immunoassay. The limited fluctuation of the theophylline levels at steady state, with twice daily administration, clearly demonstrated the marked sustained release properties of both preparations. Results indicate similar properties for the two preparations. Significant correlations between the single dose period and steady state were found for Cmax and AUC (r = 0.76 and 0.87, respectively) with one formulation, whereas this was not the case for the other (r = 0.27 and 0.49). The daily dose necessary to keep the plasma concentration within the therapeutic range of 55-110 mumole/liter varied from 7.9 to 22.9 mg/kg. Only mild side effects were recorded, but they were not correlated to the plasma theophylline concentration.


[Feeding preterm infants: application of developmental care].
Developmental care is the core concept of the preterm infant caring model. It is characterized by interaction, individualization, and evolving process. In order to enhance family competency with regard to preterm infant care, health professionals should include parents as team members when preterm infants are hospitalized so that parents can provide care helpful to facilitate preterm infant adaptation to external environmental stimulation. The preterm infant's 6-8 feedings per day represent the most frequent daily activity and an activity in which safety is essential. Potential adverse feeding responses in preterm infants such as tachypnea, cyanosis, and fatigue may stress the mother. This is a particular risk when the mother tries to increase her infant's intake volume by over-stimulating oral-motor activity. Theories and guidelines for feeding preterm infants in western societies have been evolving since the 1980s. However, little has been done in this area in Taiwan. This article addresses preterm infant feeding characteristics and cues, the application of developmental care in feeding infants, and proposes guidelines for feeding preterm infants. Healthcare professionals may reference our suggestions in feeding preterm infants and educating parents.


Differential chemosensitivity to antifolate drugs between RAS and BRAF melanoma cells.
The importance of the genetic background of cancer cells for the individual susceptibility to cancer treatments is increasingly apparent. In melanoma, the existence of a BRAF mutation is a main predictor for successful BRAF-targeted therapy. However, despite initial successes with these therapies, patients relapse within a year and have to move on to other therapies. Moreover, patients harbouring a wild type BRAF gene (including 25% with NRAS mutations) still require alternative treatment such as chemotherapy. Multiple genetic parameters have been associated with response to chemotherapy, but despite their high frequency in melanoma nothing is known about the impact of BRAF or NRAS mutations on the response to chemotherapeutic agents. Using cell proliferation and DNA methylation assays, FACS analysis and quantitative-RT-PCR we have characterised the response of a panel of NRAS and BRAF mutant melanoma cell lines to various chemotherapy drugs, amongst them dacarbazine (DTIC) and temozolomide (TMZ) and DNA synthesis inhibitors. Although both, DTIC and TMZ act as alkylating agents through the same intermediate, NRAS and BRAF mutant cells responded differentially only to DTIC. Further analysis revealed that the growth-inhibitory effects mediated by DTIC were rather due to interference with nucleotide salvaging, and that NRAS mutant melanoma cells exhibit higher activity of the nucleotide synthesis enzymes IMPDH and TK1. Importantly, the enhanced ability of RAS mutant cells to use nucleotide salvaging resulted in resistance to DHFR inhibitors. In summary, our data suggest that the genetic background in melanoma cells influences the response to inhibitors blocking de novo DNA synthesis, and that defining the RAS mutation status could be used to stratify patients for the use of antifolate drugs.


Suitability of Chinese oak silkworm eggs for the multigenerational rearing of the parasitoid Trichogramma leucaniae.
Trichogramma leucaniae is believed to be an efficient biological control agent for controlling the soybean pod borer [SPB; Leguminivora glycinivorella]. The large eggs of Chinese oak silkworm, Antheraea pernyi, are one of the best alternative host for mass production of Trichogramma. However, they are considered poor host for the growth and development of T. leucaniae. Here, we determine the feasibility of successive rearings of T. leucaniae on the large eggs for eight generations and evaluated their capacity of parasitizing SPB eggs of different ages. In the first four generations, the suitability of T. leucaniae reared on large eggs exhibited a significant increasing tendency and then decreased with the successive generations thereafter. The percentage of parasitized eggs and number of emerged adults per egg were increased from 40.0% and 10.8 adults/egg in F1 generation to 86.7% and 36.4 adults/egg in F4 generation respectively. In addition, T. leucaniae reared on A. pernyi for four generations significantly parasitized more SPB eggs regardless of egg age compared with those reared on Corcyra cephalonica eggs. These results provided useful information on the feasibility of mass production of T. leucaniae by reared for successive generations on A. pernyi large eggs.


[Pathogenesis of "Leningrad" (blockade) hypertension (60-th anniversary of the Leningrad blockade)].
Leningrad (blockade) hypertension observed in survivors of the Leningrad blockade during the World War II is a unique form of hypertension initiated and maintained pathogenetically by disturbed neuroregulation resultant from a severe psychoemotional stress. Pathogenesis of Leningrad hypertension involves mechanisms playing a key role in pathogenesis of other forms of essential hypertension. A characteristic feature of this pathogenesis is interaction of the initial and key neurogenic factor with such hypertensive factors of alimentary-dystrophic genesis as lesions of the vascular wall secondary to marked hypoproteinemia and hypovolemia. Later, in addition to hypertension the patients developed atherosclerosis, nephrosclerosis and other diseases.


Molecular cloning and physical characterization of a chromosomal hemolysin from Escherichia coli.
A chromosomal hemolysin determinant was isolated in the form of recombinant plasmid pSF4000 from a human urinary tract isolate. Escherichia coli J96. A restriction endonuclease fragment map of pSF4000 was constructed. The ampicilin resistance transposon Tn/ was used as a site-specific mutagen in conjunction with in vitro derived deletions to localize the hemolysin determinant to a 7.0-kilobase region of pSF4000. Seventeen hemolytic E. coli isolates from human urinary tract infections were found to share similar DNA sequences with J96 hemolysin sequences, using a hemolysin-specific restriction endonuclease fragment as a hybridization probe. In all 17 hemolytic E. coli strains the hemolysin gene sequences were localized to the bacterial chromosome. The hemolysin-specific gene sequences can be found only in hemolytic strains by colony blot hybridization, suggesting that in the evolutionary sense the hemolysin-specific genetic sequences have only recently been introduced into the E. coli chromosome. Evidence is also presented that in the regions neighboring the J96 chromosomal hemolysin there are additional unique DNA sequences not commonly found in other E. coli isolates.


Multiparametric MRI analysis for the identification of high intensity focused ultrasound-treated tumor tissue.
In this study endogenous magnetic resonance imaging (MRI) biomarkers for accurate segmentation of High Intensity Focused Ultrasound (HIFU)-treated tumor tissue and residual or recurring non-treated tumor tissue were identified. Multiparametric MRI, consisting of quantitative T1, T2, Apparent Diffusion Coefficient (ADC) and Magnetization Transfer Ratio (MTR) mapping, was performed in tumor-bearing mice before (n = 14), 1 h after (n = 14) and 72 h (n = 7) after HIFU treatment. A non-treated control group was included (n = 7). Cluster analysis using the Iterative Self Organizing Data Analysis (ISODATA) technique was performed on subsets of MRI parameters (feature vectors). The clusters resulting from the ISODATA segmentation were divided into a viable and non-viable class based on the fraction of pixels assigned to the clusters at the different experimental time points. ISODATA-derived non-viable tumor fractions were quantitatively compared to histology-derived non-viable tumor volume fractions. The highest agreement between the ISODATA-derived and histology-derived non-viable tumor fractions was observed for feature vector {T1, T2, ADC}. R1 (1/T1), R2 (1/T2), ADC and MTR each were significantly increased in the ISODATA-defined non-viable tumor tissue at 1 h after HIFU treatment compared to viable, non-treated tumor tissue. R1, ADC and MTR were also significantly increased at 72 h after HIFU. This study demonstrates that non-viable, HIFU-treated tumor tissue can be distinguished from viable, non-treated tumor tissue using multiparametric MRI analysis. Clinical application of the presented methodology may allow for automated, accurate and objective evaluation of HIFU treatment.


Evidence-based care and outcomes of acute stroke managed in hospital specialty units.
To assess the use of evidence-based investigations and treatments in patients with acute stroke in selected Australian hospitals and to compare management and outcomes between stroke and other types of hospital specialty unit. Retrospective, multicentre audit of hospital case files. Eight metropolitan tertiary-care hospitals from five Australian States. 300 consecutive patients from each hospital admitted between 17 September 1999 and 23 May 2001 and having a discharge diagnosis of stroke or transient ischaemic attack. Use of investigations and treatments supported by best available evidence; comparison of management and outcomes between stroke, neurology, general medical and geriatric units. 2383 patients were audited (median age, 72.7 years; 52% men); 72% had ischaemic events, and 28% haemorrhagic events. Use of investigations and treatments varied between hospitals and types of unit. Stroke units or teams cared directly for 23% of patients (range across hospitals, 0-100%). Although 47% of patients with ischaemic events presented within 3 hours of symptom onset (when thrombolysis might provide benefit), only nine (2%) received thrombolysis. Angiotensin-converting enzyme (ACE) inhibitors were given to 28% of survivors at discharge (range, 14%-38%). Stroke units were more likely to use diagnostic tests, while neurology units were more likely to prescribe heparin acutely for patients with ischaemic stroke (not recommended for patients in general), and geriatric units were less likely to discharge patients with atrial fibrillation on anticoagulation therapy. Outcomes also varied significantly between types of unit. In-hospital survival rates were 90% (stroke units), 91% (neurological units), 82% (general medical units) and 79% (geriatric units) (P < 0.001). Stroke units and neurological units sent more patients home than the other units. Stroke units also sent fewer patients to rehabilitation and had longer mean length of stay. Acute stroke care varies between Australian tertiary-care hospitals and types of specialty unit, with suboptimal use of many evidence-based interventions.


Brain tumor microvesicles: insights into intercellular communication in the nervous system.
Brain tumors are heterogeneous tumors composed of differentiated tumor cells that resemble various neural cells and a small number of multipotent cancer stem cells. These tumors modify normal cells in their environment to promote tumor growth, invasion and metastases by various ways. Recent publications show that glioblastoma cells release microvesicles that contain a select subset of cellular proteins and RNAs. These microvesicles are avidly taken up by normal cells in cell culture and can change the translational profile of these cells through delivery of tumor-derived mRNAs, which are translated into functional proteins. In addition to mRNA and proteins, microvesicles have been shown to contain microRNAs, non-coding RNAs and DNA. This commentary explores the recent advances in this novel intercellular communication route and discusses the potential physiological role of microvesicles in brain tumorigenesis.


Anti-Müllerian hormone is produced heterogeneously in primate preantral follicles and is a potential biomarker for follicle growth and oocyte maturation in vitro.
The main goals of this study were to investigate the expression of anti-Müllerian hormone (AMH) and its receptor (AMHR2) during follicular development in primates, and to evaluate the potential of AMH as a biomarker for follicle growth and oocyte maturation in vitro. The mRNA and protein expression of AMH and AMHR2 were determined using isolated follicles and ovarian sections from rhesus macaques (n = 4) by real-time PCR and immunohistochemistry, respectively. Isolated secondary follicles were cultured individually. Follicle growth and media AMH concentrations were assessed by ELISA. The mRNA expression profiles, obtained from RNA sequencing, of in vitro- and in vivo-developed antral follicles were compared. Secondary follicles from additional animals (n = 35) were cultured. Follicle growth, oocyte maturation, and media AMH concentrations were evaluated for forecasting follicular development in vitro by AMH levels. AMH immunostaining was heterogeneous in the population of preantral follicles that were also stained for AMHR2. The mRNA expression profiles were comparable between in vivo- and in vitro-developed follicles. AMH levels produced by growing follicles were higher than those of nongrowing follicles in culture. With a cutoff value of 1.40 ng/ml, 85 % of nongrowing follicles could be identified while eliminating only 5 % of growing follicles. Growing follicles that generated metaphase II-stage oocytes secreted greater amounts of AMH than did those yielding immature germinal vesicle-stage oocytes. AMH, co-expressed with AMHR2, was produced heterogeneously by preantral follicles in macaques with levels correlated positively with follicle growth and oocyte maturation. AMH may serve as a biomarker for primate follicular development in vitro.


Inhibition of pokeweed antiviral protein (PAP) by turnip mosaic virus genome-linked protein (VPg).
Pokeweed antiviral protein (PAP) from Phytolacca americana is a ribosome-inactivating protein (RIP) and an RNA N-glycosidase that removes specific purine residues from the sarcin/ricin loop of large rRNA, arresting protein synthesis at the translocation step. PAP is also a cap-binding protein and is a potent antiviral agent against many plant, animal, and human viruses. To elucidate the mechanism of RNA depurination, and to understand how PAP recognizes and targets various RNAs, the interactions between PAP and turnip mosaic virus genome-linked protein (VPg) were investigated. VPg can function as a cap analog in cap-independent translation and potentially target PAP to uncapped IRES-containing RNA. In this work, fluorescence spectroscopy and HPLC techniques were used to quantitatively describe PAP depurination activity and PAP-VPg interactions. PAP binds to VPg with high affinity (29.5 nm); the reaction is enthalpically driven and entropically favored. Further, VPg is a potent inhibitor of PAP depurination of RNA in wheat germ lysate and competes with structured RNA derived from tobacco etch virus for PAP binding. VPg may confer an evolutionary advantage by suppressing one of the plant defense mechanisms and also suggests the possible use of this protein against the cytotoxic activity of ribosome-inactivating proteins.