Characterization of the radioiodinated analogue of SCH 23390: in vitro and in vivo D-1 dopamine receptor binding studies.
A new radioiodinated molecule, 125I-SCH 38840 (previously referred to as 125I-SCH 23982), has been recently reported to be a D-1 dopamine receptor ligand. The current study confirms and expands the characterization of both the radiolabeled and unlabeled forms of this compound, as well as describing the development of an in vivo D-1 receptor binding assay utilizing the 125I-SCH 38840. The binding of 125I-SCH 38840 to rat striatal membranes, in vitro, was saturable and exhibited a KD of 1.47 nM. Competition studies using 125I-SCH 38840 exhibited a pharmacological profile consistent with the proposal that 125I-SCH 38840 was binding to the D-1 receptor. Further studies with the unlabeled SCH 38840 demonstrated that it inhibited dopamine-stimulated adenylate cyclase with a KI of 66.1 nM, indicating that SCH 38840 was acting as a D-1 antagonist. Behavioral studies demonstrated that SCH 38840 (MED = 1.0 mg/kg, s.c.) blocked conditioned avoidance responding in rats, a measurement considered predictive of anti-psychotic activity in man. In vivo binding of 125I-SCH 38840 to rat striatum following s.c. administration was specific. Peak striatal levels were observed 1 h after injection, with measurable binding observed out to 8 h post-treatment. The displacement of the in vivo binding by unlabeled standards again suggested a D-1 selective interaction. The half-life of the in vivo binding of 125I-SCH 38840 was approximately 1.25 h, and was nearly equivalent to the half-life of the anti-CAR activity of unlabeled SCH 38840. These results clearly demonstrate the D-1 nature of SCH 38840's behavioral activity and strengthen the anti-psychotic potential of a D-1 antagonist.


[Beliefs and feelings experienced by mothers of children under phototherapy].
Case study with mothers of newborn babies submitted to phototherapy held in a maternity in Fortaleza, Brazil. The purpose was identifying the mother's beliefs and feelings before the treatment. The data were collected in an interview performed with two open questions about the mother's beliefs and feelings. The result was: 36.5% of the mothers feel fear and sadness; 27.3% with sorrow and fault. Regarding beliefs, 45.5% of the mothers think that the baby feels uncomfortable; 27.3% of the mothers think that the baby is at risk of life. the mothers feel insecurity and unhappiness before the phototherapy treatment and they think that the treatment is uncomfortable for the newborn.


[Electron structure of the heme of reduced cytochrome P450 and P420 from the data of low-temperature magnetic circular dichroism].
Magnetic circular dichroism spectra (MCD) of reduced cytochromes P450 and P420 from rabbit liver microsomes have been recorded and analyzed for the 350-600 nm spectral region in the temperature interval from 2 to 290 K. The shape, intensity and temperature dependence of the MCD of reduced P450 in the Soret region are quite different from that of other high-spin ferrous hemoproteins, whose heme iron is coordinated to the imidazole of histidine (deoxymyoglobin, deoxyhemoglobin, reduced peroxidase and cytochrome c oxidase). Assuming that in the reduced P450 as well as in its CO-complex the protein-derived ligand is mercaptide (RS-) the differences can be explained by the existence of two electronic transitions in the Soret region: the common for hemoproteins pi----pi porphyrin transition and sulfur to porphyrin charge-transfer transition, p+(Sp)----eg (pi). The unusual spectral characteristics of the CO-complex of P450 have been ascribed earlier to strong configurational interaction of these two transitions. From the similarities of the Soret MCD and their temperature dependences for the reduced P420 and for other high-spin ferrous hemoproteins one can conclude that heme iron of the reduced P420 is high-spin and is coordinated to the imidazole of histidine. The zero-field splitting parameter D of the spin Hamiltonian has been estimated from the MCD temperature dependences. The obtained splitting of approximately 30 cm-1 for P450 and of approximately 10 cm-1 for P420 exceeds that for myoglobin and hemoglobin (approximately 5 cm-1).


A pulmonary rehabilitation program reduces hospitalizations in chronic obstructive pulmonary disease patients: A cost-effectiveness study.
Although pulmonary rehabilitation (PR) is recommended in patients with chronic obstructive pulmonary disease (COPD), there is a scarcity of data demonstrating the cost-effectiveness and effectiveness of PR in reducing exacerbations. A quasi-experimental study in 200 patients with COPD was conducted to determine the number of exacerbations 1 year before and after their participation in a PR program. Quality of life was measured using the COPD assessment test and EuroQol-5D. The costs of the program and exacerbations were assessed the year before and after participation in the PR program. The incremental cost-effectiveness ratio (ICER) was estimated in terms of quality-adjusted life years (QALYs). The number of admissions, length of hospital stay, and admissions to the emergency department decreased after participation in the PR program by 48.2%, 46.6%, and 42.5%, respectively (P < 0.001 for all). Results on quality of life tests improved significantly (P < 0.001 for the two tests). The cost of PR per patient and the cost of pre-PR and post-PR exacerbations were €1867.7 and €7895.2 and €4201.9, respectively. The PR resulted in a cost saving of €1826 (total, €365,200) per patient/year, and the gain in QALYs was+0.107. ICER was -€17,056. The total cost was <€20,000/QALY in 78% of patients. PR contributes to reducing the number of exacerbations in patients with COPD, thereby slowing clinical deterioration. In addition, it is cost-effective in terms of QALYs.


Adaptation to nutrient starvation in Rhizobium leguminosarum bv. phaseoli: analysis of survival, stress resistance, and changes in macromolecular synthesis during entry to and exit from stationary phase.
The nitrogen-fixing bacterium Rhizobium leguminosarum bv. phaseoli often has to survive long periods of starvation in the soil, when not in a useful symbiotic relationship with leguminous plants. We report that it can survive carbon, nitrogen, and phosphorus starvation for at least 2 months with little loss of viability. Upon carbon starvation, R. leguminosarum cells were found to undergo reductive cell division. During this period, they acquired the potential for long-term starvation-survival, levels of protein, DNA, and RNA synthesis were decreased to base levels, and pool mRNA was stabilized. The starved cells are ready to rapidly restart growth when nutrients become available. Upon addition of fresh nutrients, there is an immediate increase in the levels of macromolecular synthesis, pool mRNA destabilizes, and the cultures enter exponential growth within 5 to 8 h. The starved cells were cross-protected against pH, heat, osmotic, and oxidative shock. These results provide evidence for a general starvation response in R. leguminosarum similar to that previously found in other bacteria such as Escherichia coli and Vibrio sp.


Labelling studies for structure elucidation of a new hydroxymetabolite of tramadol.
Abstract Tramadol, racemic 1-(3-methoxyphenyl)-2-(dimethylaminomethyl)cyclohexane-1-ol, is an effective analgesic drug. Metabolites of tramadol described so far originate from O- and N-demethylation and are excreted in urine directly or after conjugation. A further metabolite was found in human liver microsome incubations and in the urine of volunteers after ingestion of tramadol. To elucidate the structure of the new metabolite, seven deuterated isotopomers of tramadol have been synthesized and ingested by volunteers. The mass spectra of the metabolites derived showed (i) that it was a hydroxy metabolite, (ii) that the hydroxy group was not located on the aromatic ring, the side chain, or the positions 2 and 6 of the cyclohexane ring, (iii) that the hydroxy-group was introduced to one of the the positions 3, 4 or 5 of the cyclohexane ring. The hydroxy metabolite was formed preferentially from the (-)-enantiomer, (1S,2S)-tramadol.


A positive feedback loop mediated by Sigma X enhances expression of the streptococcal regulator ComR.
Natural transformation is used by bacteria to take up DNA from their surroundings and incorporate it into their genomes. Streptococci do so during a transient period of competence, triggered by pheromones that they produce, secrete and sense under conditions influenced by the environment. In Streptococcus mutans, Streptococcus suis, and species of the bovis, salivarius and pyogenic groups of streptococci, the pheromone XIP is sensed by the intra-cellular regulator ComR, that in turn activates the transcription of comS, encoding the XIP precursor, and of sigX, encoding the only known alternative sigma factor in streptococci. Although induction of comR during competence has been known for more than fifteen years, the mechanism regulating its expression remains unidentified. By a combination of directional RNA-sequencing, optimal competence conditions, stepwise deletions and marker-less genome editing, we found that SigX is the missing link in overproduction of ComR. In the absence of comR induction, both sigX expression and transformation were significantly reduced. Placing comR and comS transcripts under the control of different regulators so as to form two interlocked positive feedback circuits may enable S. mutans to fine-tune the kinetics and magnitude of the competence response according to their need.


Mineralization of human premolar occlusal fissures. A quantitative histochemical microanalysis.
The mechanisms of cariogenesis in occlusal fissures remain elusive because of limited information about fissure structure and wall mineralization. The purpose of the present study was to determine the correlation between morphological patterns in occlusal fissures in human premolars and quantitative histochemical patterns of mineralization in the walls of these formations. We used scanning electron microscopy and quantitative X-ray microanalysis with the peak-to-local background ratio method and microcrystalline calcium salts as standards. We distinguished three morphological patterns of fissures in scanning electron microscopic images. The wall of the fissures was less mineralized than the control enamel in all three types of fissures. Because the fissure walls are hypomineralized, we suggest that practicing dentists should take into account the degree of mineralization when they are preparing the fissures for the application of sealant.


[Anti-hepatitis B vaccination: after the health authorities' decision].
In 1991, the WHO proposed that the antihepatitis B vaccine be included in national vaccination programs. In France, it was decided in 1994 to move on from a selective immunization scheme aimed at high-risk subjects to a general vaccination policy aimed first at adolescents in school and secondly infants. During this period, several cases of demyelinizing diseases after vaccination led to court actions and were magnified in the popular press, interfering with this new vaccination policy. On October 1, 1988, the French Secretary of Health decided to interrupt the school vaccination programs. This decision, taken as a precautionary measure, had some unexpected consequences. As had been feared by the WHO, there was an overall drop in the public's confidence in the vaccine. Health care professionals highly criticized the spectacular nature of the decision and the absence of scientific argumentation. These events lead us to raise a certain number of questions concerning the use of the public press in the management of health information, the notion of medical responsibility, and the rationality of political decisions in public health.


MicroRNA profiling of hepatocarcinogenesis identifies C19MC cluster as a novel prognostic biomarker in hepatocellular carcinoma.
Progressive hepatocarcinogenesis is a stepwise process that drives liver transformation. However, the molecular mechanisms of early liver transformation are far from clear. A role for microRNAs (miRNA) as diagnostic and prognostic factors in human tumours, including hepatocellular carcinoma (HCC), is promising. We aimed to identify novel miRNA as biomarkers for differential diagnosis and predictors of disease progression. We used a low-density array platform to profile the expression of 664 mature miRNA in a cohort of 60 hepatitis C virus-positive liver lesions representative of all stages of progressive hepatocarcinogenesis. We validated selected miRNA in two independent patient series by qPCR and we characterized the genomic status of the miRNA cluster C19MC by fluorescent in situ hybridization and copy-number variation analyses. A 18-miRNA signature distinguished cirrhosis, dysplastic nodules and HCC lesions. Four miRNAs overexpressed in HCCs belonged to chromosome 19 miRNA cluster (C19MC). Significant overexpression of C19MC in early HCC compared to dysplastic nodules could be confirmed in a second series of hepatitis B virus-related liver lesions (n = 30). In a third series of 61 HCCs, C19MC cluster was overexpressed in HCCs compared to matched cirrhotic parenchyma and regardless of the type of viral infection. High C19MC miRNA levels were correlated with poor clinico-pathological features, increased risk of tumour recurrence and shorter overall survival time. HCCs overexpressing the C19MC cluster showed genetic amplification of the corresponding locus. C19MC cluster is a novel molecular alteration characteristic of liver cancer and predictor of poor prognosis. C19MC is an attractive candidate for novel HCC therapies.


Extraction Optimization for Phenolic- and Withanolide-Rich Fractions from Withania somnifera Roots: Identification and Quantification of Withaferin A, 12-Deoxywithastromonolide, and Withanolide A in Plant Materials and Marketed Formulations Using a Reversed-Phase HPLC-Photodiode Array Detection Method.
Background: Both the roots and leaves of Withania somnifera are products of commerce. They contain active compounds of therapeutic value and mostly different withanolides. Several pharmacological activities of W. somnifera have links to one or more withanolides. The presence of phenolic compounds in extracts could play a vital role in the reduction of blood glucose levels in diabetic subjects. Objective: The present study was carried out for the selection of a solvent to prepare extracts rich in phenolics, withaferin A (WA), 12-deoxywithastromonolide (12WD), and withanolide A (WDA). A simple, rapid HPLC method was also developed for the identification and quantification of WA, 12WD, and WDA. Methods: The extraction efficiency of aqueous alcoholic solvents including hexane, chloroform, ethyl acetate, and methanol were compared for three selected withanolides and total phenolic content. The contents of WA, 12WD, and WDA and total phenolics were determined in the extracts. The quality of nine formulations containing W. sominfera were also compared in terms of the content of WA, 12WD, and WDA and total phenolics. Results: The maximum extract yield and the total withanolide and phenolic content were obtained from aqueous alcoholic compositions at 50:50 (v/v), 70:30 (v/v), and 100:0 (v/v), respectively. In the case of organic solvents, chloroform and ethyl acetate yielded the highest concentrations of phenolics and three withanolides, respectively. The total phenolic content in formulations was in the range of 1.84-3.13%, and total withanolide content showed wide variability. Conclusions: The outcome of the present investigation could be utilized for the selection of extraction solvents to prepare W. somnifera-enriched extracts and their quality monitoring by using the developed and validated HPLC-Photodiode array detection method. Highlights: A process for preparation of phenolics and withanolides (withaferin A, 12-deoxywithastramonolide and withanolide A) enriched extracts of Withania somnifera. Simple and rapid HPLC method was also developed and validated as per the ICH guidelines for identification and quantification of three major withanolides. The developed HPLC method was applied to analyze the quality of extracts and marketed herbal products (mono, as well as poly constituents). Optimized extraction process could be utilized for upscaling process development in preparation of enriched extracts from Withania somnifera, crop improvement, bio-prospection studies and quality control.


Case 269: Sacroiliac Joint Hydatid Disease.
HistoryA 20-year old woman living in Turkey presented with a 3-month history of lower back pain. She had no medical history of note and was taking no medications. Complete blood count, C-reactive protein level, sedimentation rate, and creatinine, alanine aminotransferase, and aspartate aminotransferase levels were within normal limits. Anteroposterior pelvic radiography and unenhanced pelvic CT were performed to rule out sacroiliitis. The imaging findings were abnormal, and the patient underwent contrast-enhanced sacroiliac MRI. A few days later, she underwent contrast-enhanced (100 mL iohexol, Omnipaque; GE Healthcare, Cork, Ireland) abdominal CT because of right upper quadrant pain.


Development of an AFLP based linkage map of Ler, Col and Cvi Arabidopsis thaliana ecotypes and construction of a Ler/Cvi recombinant inbred line population.
An amplified fragment polymorphism (AFLP) based linkage map has been generated for a new Landsberg erecta/ Cape Verde Islands (Ler/Cvi) recombinant inbred line (RIL) population. A total of 321 molecular PCR based markers and the erecta mutation were mapped. AFLP markers were also analysed in the Landsberg erecta/Columbia (Ler/Col) RIL population (Lister and Dean, 1993) and 395 AFLP markers have been integrated into the previous Arabidopsis molecular map of 122 RFLPs, CAPSs and SSLPs. This enabled the evaluation of the efficiency and robustness of AFLP technology for linkage analyses in Arabidopsis. AFLP markers were found throughout the linkage map. The two RIL maps could be integrated through 49 common markers which all mapped at similar positions. Comparison of both maps led to the conclusion that segregating bands from a common parent can be compared between different populations, and that AFLP bands of similar molecular size, amplified with the same primer combination in two different ecotypes, are likely to correspond to the same locus. AFLPs were found clustering around the centromeric regions, and the authors have established the map position of the centromere of chromosome 3 by a quantitative analysis of AFLP bands using trisomic plants. AFLP markers were also used to estimate the polymorphism rate among the three ecotypes. The larger polymorphism rate found between Ler and Cvi compared to Ler and Col will mean that the new RIL population will provide a useful material to map DNA polymorphisms and quantitative trait loci.


Conduct problems trajectories and psychosocial outcomes: a systematic review and meta-analysis.
There is increasing evidence that youth who follow the early onset persistent (EOP), adolescent-onset (AO) and childhood-limited (CL) trajectories of conduct problems show varying patterns of health, mental health, educational, and social outcomes in adulthood. However, there has been no systematic review and meta-analysis on outcomes associated with different conduct problems trajectories. We systematically reviewed the literature of longitudinal studies considering outcomes of three conduct problems trajectories: EOP, AO, and CL compared with individuals with low levels of conduct problems (low). We performed a series of meta-analyses comparing each trajectory to the low group for eight different outcomes in early adulthood or later. Thirteen studies met our inclusion criteria. Outcomes were mental health (depression), cannabis use, alcohol use, self-reported aggression, official records of antisocial behaviour, poor general health, poor education, and poor employment. Overall, EOP individuals showed significant higher risk of poor outcome followed by AO individuals, CL individuals, and finally participants in the low group. All conduct problems trajectories showed higher risk of poor psychosocial outcomes compared to the low group, but the magnitude of risk differed across trajectories, with a general trend for the EOP to perform significantly worse, followed by the AO and CL. Early intervention is recommended across domains to maximise likelihood of desistance from antisocial behaviour and improvement on several psychosocial outcomes.


DNA binding by 1-nitropyrene and dinitropyrenes in vitro and in vivo: effects of nitroreductase induction.
1-Nitropyrene, the predominant nitropolycyclic aromatic hydrocarbon found in diesel exhaust, is both a mutagen and a tumorigen. 1,6-Dinitropyrene is present in diesel exhaust in much smaller quantities than is 1-nitropyrene, but is much more mutagenic and carcinogenic. In an attempt to understand this difference in biological potencies, we have compared the extents to which these two nitropyrenes bind DNA in vivo. We have also determined the effect of 1-nitropyrene pretreatment upon the induction of nitroreductases and the subsequent DNA binding by both 1-nitropyrene and 1,6-dinitropyrene. In subsequent experiments, we have examined the importance of acetylation phenotype in the formation of DNA adducts from dinitropyrene in vivo. After a single intraperitoneal injection of 1-nitropyrene, covalent DNA binding could not be detected in vivo; however, 1,6-dinitropyrene formed N-(deoxyguanosin-8-yl)-1-amino-6-nitropyrene as the major DNA adduct in rat liver, kidney, urinary bladder, and mammary gland, with the highest levels being found in the bladder. The capability of liver microsomes to catalyze the oxidative metabolism of 1-nitropyrene was unchanged after treating rats with a single dose of 8 mg of 1-nitropyrene per kilogram of body weight. Cytochrome P-450, reduced nicotinamide adenine dinucleotide phosphate (NADPH2)-cytochrome P-450 reductase, and cytochrome b5 levels were also unchanged, while slight increases were detected in reduced nicotinamide adenine dinucleotide (NADH)-cytochrome b5 reductase and epoxide hydrase activities. Liver cytosolic and microsomal nitroreductase activities toward both 1-nitropyrene and 1,6-dinitropyrene were increased twofold, and cytosolic nitrosoreductase activity toward 1-nitrosopyrene and 1-nitro-6-nitrosopyrene was elevated by about 20 percent. DNA binding of both 1-nitropyrene and 1,6-dinitropyrene in vitro was twofold higher when cytosol from rats pretreated with 1-nitropyrene was used. However, pretreatment of rats with 1-nitropyrene only slightly increased the amount of in vivo DNA binding by 1,6-dinitropyrene except in the kidney, where there was a 60 percent increase. In the presence of S-acetylcoenzyme A, liver cytosol from slow-acetylator phenotype hamster strains Bio. 1.5 and 82.73 catalyzed the binding of two-to-three times more 1,8-dinitropyrene to DNA than was observed with the fast-acetylator phenotype strain Bio. 87.20. Similarly, when 1,8-dinitropyrene was administered in vivo and the extent of binding was assayed in liver, bladder, and intestinal DNA, there was more binding in strain Bio. 1.5 than in strain Bio. 87.20.(ABSTRACT TRUNCATED AT 400 WORDS)


Valuing the commons: An international study on the recreational benefits of the Baltic Sea.
The Baltic Sea provides benefits to all of the nine nations along its coastline, with some 85 million people living within the catchment area. Achieving improvements in water quality requires international cooperation. The likelihood of effective cooperation is known to depend on the distribution across countries of the benefits and costs of actions needed to improve water quality. In this paper, we estimate the benefits associated with recreational use of the Baltic Sea in current environmental conditions using a travel cost approach, based on data from a large, standardized survey of households in each of the 9 Baltic Sea states. Both the probability of engaging in recreation (participation) and the number of visits people make are modeled. A large variation in the number of trips and the extent of participation is found, along with large differences in current annual economic benefits from Baltic Sea recreation. The total annual recreation benefits are close to 15 billion EUR. Under a water quality improvement scenario, the proportional increases in benefits range from 7 to 18% of the current annual benefits across countries. Depending on how the costs of actions are distributed, this could imply difficulties in achieving more international cooperation to achieve such improvements.


Surgical experience with an implanted artificial cervical joint.
To assess the effectiveness of Cummins' artificial cervical joint, the authors reviewed the cases of 20 patients in whom the joint had been placed. A review of patients' medical records and reexamination of 18 patients were performed. The review of the surgical experience with the implantation of movable stainless-steel joints in 20 patients treated for cervical myelopathy (16 patients), cervical radiculopathy (three patients), and severe pain (one patient) indicated that the procedure is safe and well tolerated and does preserve cervical joint motion in most patients over an extended period of observation. To date, adjacent segmental symptomatic degenerative changes leading to further surgical treatment have been avoided. The joint has been placed in patients with advanced congenital and acquired cervical fusion and has been demonstrated to be stable, mobile, and biomechanically and biochemically compatible; it has shown no subsidence into adjacent bone. Wear debris has not occurred. The use of stainless steel in the cervical spine appears to be suitable for this joint replacement design.


Force controlled and teleoperated endoscopic grasper for minimally invasive surgery--experimental performance evaluation.
Minimally invasive surgery generates new user interfaces which create visual and haptic distortion when compared to traditional surgery. In order to regain the tactile and kinesthetic information that is lost, a computerized force feedback endoscopic surgical grasper (FREG) was developed with computer control and a haptic user interface. The system uses standard unmodified grasper shafts and tips. The FREG can control grasping forces either by surgeon teleoperation control, or under software control. The FREG performance was evaluated using an automated palpation function (programmed series of compressions) in which the grasper measures mechanical properties of the grasped materials. The material parameters obtained from measurements showed the ability of the FREG to discriminate between different types of normal soft tissues (small bowel, lung, spleen, liver, colon, and stomach) and different kinds of artificial soft tissue replication materials (latex/silicone) for simulation purposes. In addition, subjective tests of ranking stiffness of silicone materials using the FREG teleoperation mode showed significant improvement in the performance compared to the standard endoscopic grasper. Moreover, the FREG performance was closer to the performance of the human hand than the standard endoscopic grasper. The FREG as a tool incorporating the force feedback teleoperation technology may provide the basis for application in telesurgery, clinical endoscopic surgery, surgical training, and research.


Autoantibody activity of the immunoglobulin secreted by a follicular B-cell lymphoma.
In patients with B-cell lymphoma, only in rare cases a secreted paraprotein is found, and in very few of them an associated autoantibody activity has been demonstrated. Here we report the case of a patient with a low-grade B-cell lymphoma with a serum biclonal paraprotein (G,M)lambda and severe erythroblastopenia. Indirect immunofluorescence studies of total serum revealed cytoplasmic (Hep-2 cells) and extracellular matrix (rat tissue sections) staining, suggestive of a new specificity. After gel filtration of serum samples, only the IgM-containing fraction showed the same pattern of staining. Tumor-derived hybridomas expressed an unmutated V3-11 gene identical to that found in tumor samples and secreted an IgM immunoglobulin endowed with the same reactivity, which confirms the tumoral origin of the tissue-reactive protein. The results suggest a link between the autoimmune condition in this patient and the novel specificity displayed by the tumor-derived immunoglobulin.


Hand-held Dynamometer Measurements: Tester Strength Makes a Difference.
Three examiners with measurably different strengths and three muscle groups with distinctly different maximum force productions were used to investigate the effect of tester strength on the magnitude and reliability of hand-held dynamometer (HHD) force measurements. Each examiner measured the shoulder external rotator, elbow flexor, and knee extensor muscles of 27 healthy adults twice during two sessions (one week apart). Intrarater/intrasession, intrarater/intersession, and interrater/intrasession reliability coefficients (ICCs) were calculated. Interrater ICCs decreased in magnitude (.932 to .226) as the tested muscle groups increased in force production. Intrarater/intersession ICCs indicated a similar trend. High intrarater/intrasession and interrater/intrasession ICCs calculated from shoulder external rotator measurements indicated that HHD testing for muscle groups generating a mean maximum force of about 120 N may be reliable for clinicians with strengths equal to or exceeding those of the weakest tester in this study. Above 120 N, tester strength appeared to be a major determinant of the magnitude and reliability of the forces measured with a HHD. Further investigation into the relationship between tester strength and the intrarater and interrater reliability of HHD measurements is warranted. J Orthop Sports Phys Ther 1991;13(4):191-198.


Lactation after infant death: an analysis of Australian healthcare agencies' online health information.
Lactation is a potent signifier of maternal love and care commonly associated with early motherhood and infant survival. It is common, however, for bereaved mothers who have recently undergone miscarriage, stillbirth or infant death to produce breastmilk. Drawing on a critical feminist lens that seeks to understand how maternal subjectivities and lactation norms are constructed through public and reproductive health information, this article tests whether lactation management options after stillbirth and infant death are comprehensively covered in Australia's health organisations' online information. A qualitative directed content analysis was conducted to critique the information provided on 21 Australian websites. Information extracted from websites was compared to a 'best-practice' Lactation After Infant Death (AID) Framework developed by the research team for the review. We found a notable absence of comprehensive lactation management information targeted directly to bereaved mothers. Moreover, the most common lactation option presented for women without a living infant was lactation suppression. This dearth of appropriate and comprehensive lactation information curtails maternal subjectivies and diverse lactation practices and further isolates women dealing with the painful contradictions of lactation after loss.


Macrophages, chemokines and neuronal injury in HIV-1-associated dementia.
Human immunodeficiency virus type-one (HIV- 1)-associated dementia (HAD) is manifested as a spectrum of behavioral, motor and cognitive dysfunctions. The disorder commonly occurs during late stage HIV disease and remains an important complication despite highly active antiretroviral therapies. A metabolic encephalopathy, fueled by neurotoxic secretions from brain mononuclear phagocytes (MP) (macrophages and microglia) underlies HIV- I neuropathogenesis. One pivotal question, however, is how brain MP evolve from neurotrophic to neurotoxic cells. The interplay between the virus, the macrophage and the neuron has just recently begun to be unraveled. Along with a multitude of other MP secretory products, chemokines effect neuronal function by engaging neuronal receptors then activating pathways that alter synaptic transmission, cell growth, injury and protection. Both neurons and glia secrete chemokines. Interestingly, HIV-1 and its gene products can mimic chemokine neuronal signaling by binding to neuronal chemokine receptors or by other non-specific mechanisms. The elucidation of mechanisms involved in chemokine-mediated neural compromise will likely provide unique insights into the pathogenesis and treatment, not only of HAD, but of a wide range of neurodegenerative disorders.


Postoperative C-reactive protein/albumin ratio is a biomarker of risk of recurrence and need for adjuvant chemotherapy for stage III colorectal cancer.
Adjuvant chemotherapy is generally recommended for patients with stage III colorectal cancer. Even with adjuvant chemotherapy, 20-30% of such patients develop recurrences; the risk factors for recurrence are currently unclear. The preoperative systemic inflammation index has been linked to poor prognoses in patients with colorectal cancer; however, the relationship between postoperative systemic inflammation index and recurrence is unclear. We aimed to evaluate the association between preoperative and postoperative systemic inflammation indexes and recurrence in patients with stage III colorectal cancer. The following laboratory data of 133 patients with stage III colorectal cancer were analyzed: preoperative and postoperative C-reactive protein/albumin ratios (CAR); neutrophil to lymphocyte ratios (NLR); and platelet to lymphocyte ratios (PLR) and their relationships with recurrence analyzed. The optimal cutoff values for systemic inflammation indexes were determined by examining receiver operating characteristic curves. Multivariate analyses indicated that N-stage, postoperative complications, preoperative NLR, and postoperative CAR were independent predictors of recurrence-free survival (RFS). Postoperative CAR was also an independent predictor of overall survival (OS). Patients with postoperative CAR ≥ 0.035 who did not receive adjuvant chemotherapy had shorter RFS and OS than those who did. There were no significant differences in RFS and OS between patients with postoperative CAR < 0.035 who did and did not receive adjuvant chemotherapy. Postoperative CAR is strongly associated with poor prognosis in patients with stage III colorectal cancer and is a useful biomarker for determining whether adjuvant chemotherapy should be administered.


A rare case of fatal primary cutaneous mucinous carcinoma of the scalp with multiple in-transit and pulmonary metastases.
Primary cutaneous mucinous carcinoma is a rare neoplasm derived from the sweat glands. It is usually located in the head and neck region, with the eyelids being the most common site of presentation. Recurrence following primary excision is common but metastasis is rare. We report a patient presenting with rapidly progressive cutaneous mucinous carcinoma on the right parietal scalp. Systemic work-up failed to reveal an occult primary source. The tumor was treated with Mohs micrographic surgery with clear margins. However, within two months, the patient developed new lesions on the scalp both adjacent to and separate from the original previously excised area, as well as metastasis to the right parotid gland. The patient received adjuvant radiation therapy both to the scalp lesions and to the parotid gland with apparent control of locoregional disease. However, two weeks after completing radiation therapy, the patient was found to have developed pulmonary metastases and died soon thereafter. Primary mucinous carcinoma arising in the skin is rare and usually follows an indolent course. We report the rare occurrence of a mucinous carcinoma of the scalp that developed multiple cutaneous tumors caused by in-transit metastases, as well as parotid gland metastases and eventual fatal pulmonary metastases.


Pilot feasibility and acceptability study evaluating use of group CBT-I in improving sleep and fatigue in older adults.
This paper describes a pilot study, in a small sample of older adults, designed to ascertain the feasibility, acceptability and potential effectiveness of group Cognitive Behavioural Therapy For Insomnia (CBT-I). Eleven older adults participated in a 3-week group CBT-I intervention with pre- and post-intervention outcome measures of sleep and fatigue. Acceptability was measured via post-intervention focus groups. Feasibility was measured via recruitment, retention and completion data. Subjective sleep outcomes were measured pre- and post-intervention using the Pittsburgh Sleep Quality Index. Fatigue was measured using the Fatigue Severity Index. Objective sleep outcome measures were obtained via pre- and post-intervention wrist actigraphy. Feasibility and acceptability were confirmed in the pilot study. Retention and completion levels were high, with participants largely positive in the focus group feedback. Preliminary sleep outcome data supported the potential effectiveness of the intervention in significantly improving several measures of sleep and fatigue including a three-point reduction in the Global PSQI scores, an increase in total sleep time of almost an hour per night and these results were mirrored by a significant reduction of nine points in the overall measure of fatigue severity. A group CBT-I intervention is a low-cost, low-risk intervention which improves subjective and objective measures of sleep in older adults. These positive sleep outcomes are translated into significantly decreased levels of fatigue. Future research should focus on a larger sample size with a randomised controlled trial design.


Weight reduction interventions for persons with a chronic illness: findings and factors for consideration.
Obesity in persons with a concomitant chronic illness poses complex issues relating to the choice of appropriate interventions. More recent emphasis on modification of risk factors has resulted in the need to prescribe complex therapeutic regimens with multiple treatment goals. The traditional approach to weight reduction in such persons has been nutrition education. Studies have shown, however, that knowledge alone does not translate into self-care behaviors that in turn result in weight loss and weight maintenance. Although the latter outcomes continue to be primary goals of therapy in obese individuals with a chronic illness, improvement in the physiologic parameters associated with the illness is also a desired outcome. Behavior therapy and group support appear to be enabling factors that go beyond knowledge to facilitate behavior change and subsequent changes in health-related indexes. This article describes various approaches to the problem of combined interventions for patients education and weight reduction. Findings and factors are discussed about whether the primary goal of weight reduction interventions for persons with a chronic illness should focus on pounds lost or improvement in metabolic or physiologic status.


Expansion of a regional ST-segment-elevation myocardial infarction system to an entire state.
Despite national guidelines calling for timely coronary artery reperfusion, treatment is often delayed, particularly for patients requiring interhospital transfer. One hundred nineteen North Carolina hospitals developed coordinated plans to rapidly treat patients with ST-segment-elevation myocardial infarction according to presentation: walk-in, ambulance, or hospital transfer. A total of 6841 patients with ST-segment-elevation myocardial infarction (3907 directly presenting to 21 percutaneous coronary intervention hospitals, 2933 transferred from 98 non-percutaneous coronary intervention hospitals) were treated between July 2008 and December 2009 (age, 59 years; 30% women; 19% uninsured; chest pain duration, 91 minutes; shock, 9.2%). The rate of patients not receiving reperfusion fell from 5.4% to 4.0% (P=0.04). Treatment times for hospital transfer patients substantially improved. First-hospital-door-to-device time for hospitals that adopted a "transfer for percutaneous coronary intervention" reperfusion strategy fell from 117 to 103 minutes (P=0.0008), whereas times at hospitals with a mixed strategy of transfer or fibrinolysis fell from 195 to 138 minutes (P=0.002). Median door-to-device times for patients presenting directly to PCI hospitals fell from 64 to 59 minutes (P<0.001). Emergency medical services-transported patients were most likely to reach door-to-device goals, with 91% treated within 90 minutes and 52% being treated with 60 minutes. Patients treated within guideline goals had a mortality of 2.2% compared with 5.7% for those exceeding guideline recommendations (P<0.001). Through extension of regional coordination to an entire state, rapid diagnosis and treatment of ST-segment-elevation myocardial infarction has become an established standard of care independently of healthcare setting or geographic location.


GSTM1, GSTT1, and GSTP1 polymorphisms and risk of advanced colorectal adenoma.
Cigarette smoking is a risk factor for colon adenoma. The glutathione S-transferase enzymes are involved in the detoxification of carcinogenic compounds including those found in tobacco smoke, and thus, may be important modifiers of individual risk of developing this disease. We examined the prevalence of GSTM1 and GSTT1 gene deletions, and two GSTP1 polymorphisms in 772 cases with advanced colorectal adenomas (>1 cm, villous elements or high-grade dysplasia) of the distal colon (descending or sigmoid colon or rectum) and 777 sigmoidoscopy negative controls enrolled in the screening arm of the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial. Epidemiologic data on smoking was collected by self-administered questionnaire and DNA was extracted from whole blood or buffy coat. For GSTM1 and GSTT1, we used a newly developed TaqMan-based assay capable of discriminating heterozygous (+/-) individuals from those with two active alleles (+/+) and homozygous deletions (-/-). For GSTP1, the I105V and the A114V substitutions were identified using end point 5' nuclease assays (TaqMan). Adjusted odds ratios (OR) and 95% confidence intervals (95% CI) were determined using unconditional logistic regression, controlling for age, race, and gender. Advanced adenoma risk was increased in current/former smokers (OR, 1.4; 95% CI, 1.2-1.8). Risks were decreased in subjects with > or =1 inactive GSTM1 alleles (OR, 0.6; 95% CI, 0.4-0.9); and the association was independent of smoking status (P interaction = 0.59). Having > or =1 inactive GSTT1 allele was associated with increased risk among smokers (OR, 1.4; 95% CI, 1.1-1.9; P trend = 0.02) but not among never smokers (OR, 0.9; 95% CI, 0.6-1.3) and a significant interaction between smoking and genotype was observed (P interaction = 0.05). In summary, this is the first study to report associations between colorectal adenomas and GSTM1 wild-type and GSTT1 null allele among smokers. These findings only became apparent using a newly developed assay able to distinguish heterozygous from wild-type individuals. Our data provide evidence that phenotypic differences between these two groups exist.


Chondroclasts are mature osteoclasts which are capable of cartilage matrix resorption.
Multinucleated cells termed chondroclasts have been observed on the deep surface of resorbed hyaline cartilage but the relationship of these cells to macrophages and osteoclasts and their role in rheumatoid arthritis (RA) and other arthritic conditions is uncertain. Multinucleated cells in RA and other arthritic conditions showing evidence of cartilage resorption were characterised immunohistochemically for expression of macrophage/osteoclast markers. Mature human osteoclasts formed from circulating monocytes and tissue macrophages were cultured for up to 4 days on slices of human cartilage and glycosaminoglycan (GAG) release was measured. Multinucleated cells resorbing unmineralised cartilage were seen in osteoarthritis, RA, septic arthritis, avascular necrosis and in four cases of giant cell tumour of bone that had extended through the subchondral bone plate. Chondroclasts expressed an osteoclast-like phenotype (TRAP+, cathepsin K+, MMP9+, CD14-, HLA-DR-, CD45+, CD51+ and CD68+). Both macrophages and osteoclasts cultured on cartilage released GAG. These findings indicate that chondroclasts have an osteoclast-like phenotype and that mature human osteoclasts are capable of cartilage matrix resorption. Resorption of unmineralised subchondral cartilage by chondroclasts and macrophages can be a feature of joint destruction in inflammatory and non-inflammatory arthropathies as well as inflammatory and neoplastic subchondral bone lesions.


Designing the selenium and bladder cancer trial (SELEBLAT), a phase lll randomized chemoprevention study with selenium on recurrence of bladder cancer in Belgium.
In Belgium, bladder cancer is the fifth most common cancer in males (5.2%) and the sixth most frequent cause of death from cancer in males (3.8%). Previous epidemiological studies have consistently reported that selenium concentrations were inversely associated with the risk of bladder cancer. This suggests that selenium may also be suitable for chemoprevention of recurrence. The SELEBLAT study opened in September 2009 and is still recruiting all patients with non-invasive transitional cell carcinoma of the bladder on TURB operation in 15 Belgian hospitals. Recruitment progress can be monitored live at http://www.seleblat.org. Patients are randomly assigned to selenium yeast (200 μg/day) supplementation for 3 years or matching placebo, in addition to standard care. The objective is to determine the effect of selenium on the recurrence of bladder cancer. Randomization is stratified by treatment centre. A computerized algorithm randomly assigns the patients to a treatment arm. All study personnel and participants are blinded to treatment assignment for the duration of the study. The SELEnium and BLAdder cancer Trial (SELEBLAT) is a phase III randomized, placebo-controlled, academic, double-blind superior trial. This is the first report on a selenium randomized trial in bladder cancer patients. ClinicalTrials.gov identifier: NCT00729287.


Voglibose administration before the evening meal improves nocturnal hypoglycemia in insulin-dependent diabetic patients with intensive insulin therapy.
Nocturnal hypoglycemia is one of the serious complications of intensive insulin therapy in patients with insulin-dependent diabetes mellitus (IDDM; type 1 DM). We assessed the effect of voglibose (alpha-glucosidase inhibitor) administration before the evening meal on nocturnal hypoglycemia in IDDM patients with intensive insulin therapy. Ten IDDM patients received 0.3 mg voglibose just before the evening meal for 5 days. The diet and insulin regimen were not changed throughout the study. Nocturnal plasma glucose levels (10 PM, 3 AM, and 7 AM) were studied in these patients before and during voglibose administration. Blood glucose levels were measured at 3 AM before and during voglibose treatment. The mean plasma glucose level at 3 AM was 3.4+/-0.4 mmol/L before voglibose treatment and 7.3+/-1.0 mmol/L during treatment. Plasma glucose at 3 AM was elevated in 9 of 10 patients with voglibose. The decrease in plasma glucose from 10 PM to 3 AM was 6.5+/-0.8 mmol/L before voglibose administration but 3.2+/-0.9 mmol/L during treatment (P < .01). The hypoglycemia rate was 52% (17 of 33 nights) before voglibose administration but only 9.1% (3 of 33 nights) during treatment. We conclude that voglibose administration before the evening meal improves nocturnal hypoglycemia in IDDM patients with intensive insulin therapy.


The emergence of antigenic variants is a rare event in long-term visna virus infection in vivo.
Six sheep persistently infected with visna virus were studied for 4 1/2 to 5 3/4 years until they became ill. Virus was isolated at intervals from peripheral blood leukocytes and cerebrospinal fluid (CSF), and at the time of sacrifice from various parts of the brain and the lungs. Both brain and lungs showed lesions typical of advanced visna/maedi. All the sheep formed antibodies in sera and CSF. Virus isolates from each sheep were tested in neutralization tests against sera and CSF collected from the same animal. In one sheep all isolates were found to be identical to the inoculated virus by this test. In each of the other sheep an antigenic variant emerged from 1 to 3 years after inoculation and remained in circulation even after the formation of autologous antibodies. In one case a variant was isolated from the lungs, whereas in all cases the virus isolated from the brain was identical to the inoculated virus. The results show that antigenic variants are rare in visna and do not seem to have a role in the pathogenesis of the disease.


Effect of compression paddle tilt correction on volumetric breast density estimation.
For the acquisition of a mammogram, a breast is compressed between a compression paddle and a support table. When compression is applied with a flexible compression paddle, the upper plate may be tilted, which results in variation in breast thickness from the chest wall to the breast margin. Paddle tilt has been recognized as a major problem in volumetric breast density estimation methods. In previous work, we developed a fully automatic method to correct the image for the effect of compression paddle tilt. In this study, we investigated in three experiments the effect of paddle tilt and its correction on volumetric breast density estimation. Results showed that paddle tilt considerably affected accuracy of volumetric breast density estimation, but that effect could be reduced by tilt correction. By applying tilt correction, a significant increase in correspondence between mammographic density estimates and measurements on MRI was established. We argue that in volumetric breast density estimation, tilt correction is both feasible and essential when mammographic images are acquired with a flexible compression paddle.


Effects of ion channel blockade on the distribution of Na, K, Ca and other elements in oxygen-glucose deprived CA1 hippocampal neurons.
The pathophysiology of brain ischemia and reperfusion injury involves perturbation of intraneuronal ion homeostasis. To identify relevant routes of ion flux, rat hippocampal slices were perfused with selective voltage- or ligand-gated ion channel blockers during experimental oxygen-glucose deprivation and subsequent reperfusion. Electron probe X-ray microanalysis was used to quantitate water content and concentrations of Na, K, Ca and other elements in morphological compartments (cytoplasm, mitochondria and nuclei) of individual CA1 pyramidal cell bodies. Blockade of voltage-gated channel-mediated Na+ entry with tetrodotoxin (1 microM) or lidocaine (200 microM) significantly reduced excess intraneuronal Na and Ca accumulation in all compartments and decreased respective K loss. Voltage-gated Ca2+ channel blockade with the L-type antagonist nitrendipine (10 microM) decreased Ca entry and modestly preserved CA1 cell elemental composition and water content. However, a lower concentration of nitrendipine (1 microM) and the N-, P-subtype Ca2+ channel blocker omega-conotoxin MVIIC (3 microM) were ineffective. Glutamate receptor blockade with the N-methyl-D-aspartate (NMDA) receptor-subtype antagonist 3-(2-carboxypiperazin-4-yl) propyl-1-phosphonic acid (CPP; 100 microM) or the alpha-amino-3-hydroxy-5-methyl-4-isoazole propionic acid (AMPA) receptor subtype blocker 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 10 microM/100 microM glycine) completely prevented Na and Ca accumulation and partially preserved intraneuronal K concentrations. Finally, the increase in neuronal water content normally associated with oxygen-glucose deprivation/reperfusion was prevented by Na+ channel or glutamate receptor blockade. Results of the present study demonstrate that antagonism of either postsynaptic NMDA or AMPA glutaminergic receptor subtypes provided nearly complete protection against ion and water deregulation in nerve cells subjected to experimental ischemia followed by reperfusion. This suggests activation of ionophoric glutaminergic receptors is involved in loss of neuronal osmoregulation and ion homeostasis. Na+ channel blockade also effectively diminished neuronal ion and water derangement during oxygen-glucose deprivation and reperfusion. Prevention of elevated Nai+ levels is likely to provide neuroprotection by decreasing presynaptic glutamate release and by improving cellular osmoregulation, adenosine triphosphate utilization and Ca2+ clearance. Thus, we suggest that voltage-gated tetrodotoxin-sensitive Na+ channels and glutamate-gated ionotropic NMDA or AMPA receptors are important routes of ion flux during nerve cell injury induced by oxygen-glucose deprivation/reperfusion.


Shifts in the Clonal Distribution of Methicillin-Resistant Staphylococcus aureus in Kuwait Hospitals: 1992-2010.
As the epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) is constantly changing globally, determining the prevailing MRSA clones in a local healthcare facility is important for better management of infections. This study investigated clonal composition and distribution of MRSA isolates in Kuwait's hospitals using a combination of molecular typing methods. In total, 400 non-repeat MRSA isolates were obtained between 1992 and 2010 in 13 public hospitals and were characterized using antibiogram, SCCmec typing, spa typing, and multilocus-sequence typing. Clonal assignment and detection of virulence factors and antibiotic resistance genes were performed by DNA microarray. The isolates were resistant to kanamycin (74.2%), erythromycin (69.5%), tetracycline (66.7%), gentamicin (61%), ciprofloxacin, (61%), fusidic acid (53.5%), clindamycin (41.5%), high-level mupirocin resistance (5.2%) and carried aphA3, aacA-aphD, ermA, ermC, mupA, tetK, tetM, fusC and far1. Molecular typing revealed 31 different MRSA clones consisting of ST239-MRSA-III (52.2%), ST22-MRSA-IV (9.2%), ST80-MRSA-IV (7.5%), ST5-MRSA-II/IV/V/VI (6.5%), ST30-MRSA-IV (3.5%), ST241-MRSA-III (2.7%), ST6-MRSA-IV (2.2%), ST36-MRSA-II (2%) and ST772-MRSA-V (1.75%). The isolates differed in the carriage of genes for enterotoxins, Panton-Valentine leukocidin (PVL), toxic shock syndrome toxin (tst-1), arginine catabolic mobile element (ACME) and exfoliative toxins. The number of clones increased from one (ST239-III-t037) in 1992 to 30 in 2010 including ST8-IV-t008 [PVL+] [ACME+] (USA300), ST772-V (Bengal Bay clone) and ST2816 identified for the first time in Kuwait. The study revealed that the MRSA isolates belonged to diverse clones that changed in numbers and diversity overtime. Although ST239-MRSA-III, a healthcare-associated clone remained the dominant MRSA clone overtime, the newly emerged clones consisted mostly of community-associated.


Prevalence, Incidence and Ecological Determinants of Diabetic Retinopathy in Iran: Systematic Review and Meta-analysis.
To estimate the pooled prevalence and incidence of diabetic retinopathy (DR) in Iran and to investigate their correlations with the Human Development Index (HDI), healthcare access (i.e., density of specialists and sub-specialists), and methodological issues. Electronic databases such as PubMed, Embase, Scopus, Web of Science, Google Scholar, and local databases were searched for cohort and cross-sectional studies published prior to January 2018. Prevalence and incidence rates of DR were extracted from January 2000 to December 2017 and random effects models were used to estimate pooled effect sizes. The Joanna Briggs Institute critical appraisal tool was applied for quality assessment of eligible studies. A total of 55,445 participants across 33 studies were included. The pooled prevalence (95% CI) of DR in diabetic clinics (22 studies), eye clinics (4 studies), and general population (7 studies) was 31.8% (24.5 to 39.2), 57.8% (50.2 to 65.3), and 29.6% (22.6 to 36.5), respectively. It was 7.4% (3.9 to 10.8) for proliferative DR and 7.1% (4.9 to 9.4) for clinically significant macular edema. The heterogeneity of individual estimates of prevalence was highly significant. HDI ( P < 0.001), density of specialists ( P = 0.004), subspecialists ( P < 0.001), and sampling site ( P = 0.041) were associated with heterogeneity after the adjustment for type of DR, duration of diabetes, study year, and proportion of diabetics with controlled HbA1C. Human development and healthcare access were correlated with the prevalence of DR. Data were scarce on the prevalence of DR in less developed provinces. Participant recruitment in eye clinics might overestimate the prevalence of DR.


Understanding the spectacular failure of DNA barcoding in willows (Salix): does this result from a trans-specific selective sweep?
Willows (Salix: Salicaceae) form a major ecological component of Holarctic floras and consequently are an obvious target for a DNA-based identification system. We surveyed two to seven plastid genome regions (~3.8 kb; ~3% of the genome) from 71 Salix species across all five subgenera, to assess their performance as DNA barcode markers. Although Salix has a relatively high level of interspecific hybridization, this may not sufficiently explain the near complete failure of barcoding that we observed: only one species had a unique barcode. We recovered 39 unique haplotypes, from more than 500 specimens, that could be partitioned into six major haplotype groups. A unique variant of group I (haplotype 1*) was shared by 53 species in three of five Salix subgenera. This unusual pattern of haplotype sharing across infrageneric taxa is suggestive of either a massive nonrandom coalescence failure (incomplete lineage sorting), or of repeated plastid capture events, possibly including a historical selective sweep of haplotype 1* across taxonomic sections. The former is unlikely as molecular dating indicates that haplotype 1* originated recently and is nested in the oldest major haplotype group in the genus. Further, we detected significant non-neutrality in the frequency spectrum of mutations in group I, but not outside group I, and demonstrated a striking absence of geographical (isolation by distance) effects in the haplotype distributions of this group. The most likely explanation for the patterns we observed involves recent repeated plastid capture events, aided by widespread hybridization and long-range seed dispersal, but primarily propelled by one or more trans-species selective sweeps.


Varying views of life among people with long-term mental illness.
The purpose of this study was to explore views of life among people with long-term mental illnesses. The participants' possible cognitive inability to express such views dictated a research design that was both fit for purpose and respectful of their integrity. The study, based on an ethnographic framework, involved photographs and interviews with five women and three men. The participants were the photographers, as well as the authors of their own narratives, and the photographs served as a starting point for the interviews. The interview material was analysed in terms of the phases of interpretation. Four main themes were identified: 'thoughts about God and the meaning of life and death', 'the meaning of relationships with others', 'how animals give meaning to life without demands' and 'the symbolic bearing of objects on life'. These four themes represent key existential issues among people with long-term mental illnesses, but they lack confidants to share or discuss these matters with.


Delivery of a Protease-Activated Cytolytic Peptide Prodrug by Perfluorocarbon Nanoparticles.
Melittin is a cytolytic peptide derived from bee venom that inserts into lipid membranes and oligomerizes to form membrane pores. Although this peptide is an attractive candidate for treatment of cancers and infectious processes, its nonspecific cytotoxicity and hemolytic activity have limited its therapeutic applications. Several groups have reported the development of cytolytic peptide prodrugs that only exhibit cytotoxicity following activation by site-specific proteases. However, systemic administration of these constructs has proven difficult because of their poor pharmacokinetic properties. Here, we present a platform for the design of protease-activated melittin derivatives that may be used in conjunction with a perfluorocarbon nanoparticle delivery system. Although native melittin was substantially hemolytic (HD50: 1.9 μM) and cytotoxic (IC50: 2.4 μM), the prodrug exhibited 2 orders of magnitude less hemolytic activity (HD50: > 100 μM) and cytotoxicity (IC50: > 100 μM). Incubation with matrix metalloproteinase-9 (MMP-9) led to cleavage of the prodrug at the expected site and restoration of hemolytic activity (HD50: 3.4 μM) and cytotoxicity (IC50: 8.1 μM). Incubation of the prodrug with perfluorocarbon nanoparticles led to stable loading of 10,250 peptides per nanoparticle. Nanoparticle-bound prodrug was also cleaved and activated by MMP-9, albeit at a fourfold slower rate. Intravenous administration of prodrug-loaded nanoparticles in a mouse model of melanoma significantly decreased tumor growth rate (p = 0.01). Because MMPs and other proteases play a key role in cancer invasion and metastasis, this platform holds promise for the development of personalized cancer therapies directed toward a patient's individual protease expression profile.


[Infantile acro-localized papulovesicular syndrome].
Papular acrodermatitis (Gianotti-Crosti syndrome) was seen in a six-year-old girl. The disease was marked by the characteristic triad of a papular-vesicular rash, lymphadenopathy and liver damage. Serological findings suggest an infection with Epstein-Barr virus as the causative factor. In such cases hepatitis-B induced papular eruptive acrodermatitis should be considered in differential diagnosis.


Self-Esteem Group: Useful Intervention for Inpatients with Anorexia Nervosa?
Low self-esteem is a common feature in Anorexia Nervosa (AN) and has been hypothesised to act as a predisposing, precipitating, and perpetuating factor. The aim of this study was to assess the effectiveness of a Cognitive Behavioural Therapy (CBT)-based self-esteem group in a naturalistic setting of patients with AN in an inpatient treatment programme. Included in this study were 119 female patients diagnosed with AN, with all participants completing self-report questionnaires before and after the intervention. The group consisted of five to six weekly sessions. The self-esteem group led to a statistically significant improvement in self-esteem, which could not be explained by an increase in BMI alone, suggesting that the group is facilitating positive changes within an AN group. The group also had a small effect on improving patients self-perceived ability to change. These findings suggest that the brief self-esteem group has some benefits in improving patients' self-esteem/self-efficacy and should be replicated in the future with a control condition to confirm findings.


Fractalkine expression on human renal tubular epithelial cells: potential role in mononuclear cell adhesion.
Fractalkine (CX3CL1) is a transmembrane molecule with a CX3C chemokine domain attached to an extracellular mucin stalk which can induce both adhesion and migration of leucocytes. Mononuclear cell infiltration at renal tubular sites and associated tubular epithelial cell damage are key events during acute renal inflammation following renal allograft transplantation. Using northern and Western blot analysis, we have demonstrated the expression of fractalkine message and protein by renal tubular epithelial cells in vitro. The expression was up-regulated by TNF-alpha, a key proinflammatory cytokine in acute rejection. Investigation of surface expression of fractalkine on cultured proximal tubular epithelial cells revealed only a subpopulation of positively staining cells. Immunohistochemistry revealed that only a proportion of tubules in renal allograft biopsies showed induction of fractalkine expression. Studies using a static model of adhesion demonstrated CX3CR1/fractalkine interactions accounted for 26% of monocytic THP-1 cell and 17% of peripheral blood natural killer cell adhesion to tubular epithelial cells, suggesting that fractalkine may have a functional role in leucocyte adhesion and retention, at selected tubular sites in acute renal inflammation. Thus, fractalkine blockade strategies could reduce mononuclear cell mediated tubular damage and improve graft survival following kidney transplantation.


Emissive nanoparticles from pyridinium-substituted tetraphenylethylene salts: imaging and selective cytotoxicity towards cancer cells in vitro and in vivo by varying counter anions.
Chemotherapeutics specifically targeting cancer cells without damaging healthy cells is the long-awaited goal of cancer treatment. In this paper, a series of nanoparticles (NanoTPES 1-4) assembled from pyridinium-substituted tetraphenylethylene salts were synthesized and investigated both in vitro and in vivo for this purpose. By changing the counter anions, NanoTPES 1-4 exhibit tunable emission colors, sizes and surface charges. NanoTPES 2 and 3 with tetraphenyl borate and tetra(4-chlorophenyl) borate as the respective anions selectively imaged and targeted mitochondria in cancer cells. Accordingly, these two nanoparticles specifically kill cancer cells with minimal effect on normal cells. Such selective cytotoxicity was attributed to the change of membrane potential and inhibition of ATP synthesis in the mitochondria of cancer cells. Furthermore, both NanoTPES 2 and 3 exhibited efficient tumor accumulation and tumor growth inhibition in vivo, with negligible systemic toxicity.


Uncontrolled Hypertension, Treatment, and Predictors among Hypertensive Out-Patients Attending Primary Health Facilities in Johannesburg, South Africa.
Hypertension is a poorly controlled risk factor for cardiovascular disease in South Africa, particularly among patients receiving care in the public sector who are mostly from low socioeconomic backgrounds. This cross-sectional study investigated uncontrolled hypertension, treatment, and predictors among hypertensive out-patients attending primary health care facilities in Johannesburg, South Africa. The WHO STEPwise approach to the surveillance of non-communicable diseases was used to collect data, including sociodemographic and lifestyle factors, health status, and measurements for anthropometry and blood pressure along with self-reported adherence to treatment, estimated through the general medication adherence scale. Uncontrolled hypertension was defined as systolic blood pressure ≥140 mmHg and/or diastolic blood pressure ≥90 mmHg in diagnosed patients receiving anti-hypertensive treatment. Overweight and obesity were defined as a body mass index ≥25 and ≥30 kg/m2, respectively. Logistic regression models were used to assess the predictors of uncontrolled hypertension. Four hundred (n = 400) hypertensive out-patients (mean age: 50 ± 8 years) participated in this study, with most living in poor sociodemographic environments. The prevalence rate of uncontrolled hypertension was 57%. Obesity (62% vs. 42%, p ≤ 0.0001), salt consumption (90% vs. 55%, p ≤ 0.0001), alcohol intake (42% vs. 19%, p ≤ 0.0001), a smoking habit (23% vs. 4%, p ≤ 0.0001), alternative medicine use (51% vs. 40%, p = 0.043), and comorbidities (64% vs. 36%, p ≤ 0.0001) were higher in the uncontrolled group than the controlled group, whereas the prevalence of physical activity (38% vs. 15%, p ≤ 0.0001) was high in the controlled group vs. the uncontrolled. Overall, 85% of the patients moderately adhered to treatment, only 2% exhibited high adherence, and 13% demonstrated low adherence; over half of the patients received tri-therapy treatment. The predictors of uncontrolled hypertension are a number of prescribed antihypertensive therapies [adjusted odds ration = 2.39; 95% confidence interval: 1.48-3.87], treatment adherence [0.46; 0.21-0.97], salt consumption [28.35; 7.87-102.04], physical activity [0.22; 0.13-0.37], current alcohol use [2.10; 1.22-3.61], and current cigarette smoking [4.79; 1.88-12.18]. The high prevalence of uncontrolled hypertension in this study suggests a need to optimize prescriptions, adherence to BP-lowering medications, and lifestyle modifications. The management of comorbidities such as diabetes could offer considerable benefits in controlling blood pressure.


A randomized controlled trial examining the efficacy of an internet-based cognitive behavioral therapy program for adolescents with anxiety disorders.
Anxiety disorders are highly prevalent in adolescence, but access to health care services is limited and only few receive professional help. Internet-based cognitive behavioral therapy (ICBT) has been proposed to increase accessibility and reduce costs of treatment. The study evaluated the efficacy of a Danish version of the guided ICBT program ChilledOut Online, developed at the Centre for Emotional Health, Macquarie University, Australia. At the Centre for Psychological Treatment of Children and Adolescents, Aarhus University, Denmark, a randomized controlled trial was conducted with 70 adolescents (13-17 years) with anxiety disorders according to DSM-IV. Participants were randomly assigned to a 14-weeks therapist-guided ICBT or to a waitlist condition. Outcomes were evaluated post-treatment and at 3- and 12-month follow-up. At post-treatment, the ICBT group significantly outperformed the waitlist condition with moderate to large between-group effect sizes on diagnostic severity and anxiety symptoms rated by clinicians, and by adolescents and their parents. Forty percent of adolescents in ICBT were free of their primary diagnosis compared to 16% in the waitlist condition. Treatment gains were maintained at 3- and 12-month follow-up. Results of the study provide support for the efficacy of guided ICBT for adolescents with anxiety disorders. ClinicalTrials.gov: NCT02535403.


Reovirus-induced apoptosis is preceded by increased cellular calpain activity and is blocked by calpain inhibitors.
The cellular pathways of apoptosis have not been fully characterized; however, calpain, a cytosolic calcium-activated cysteine protease, has been implicated in several forms of programmed cell death. Reoviruses induce apoptosis both in vitro and in vivo and serve as a model for studying virus-induced cell death. We investigated the potential role of calpain in reovirus-induced apoptosis in vitro by measuring calpain activity as well as evaluating the effects of calpain inhibitors. L929 cells were infected with reovirus type 3 Abney (T3A), and calpain activity, measured as cleavage of the fluorogenic calpain substrate Suc-Leu-Leu-Val-Tyr-AMC, was monitored. There was a 1.6-fold increase in calpain activity in T3A-infected cells compared to mock-infected cells; this increase was completely inhibited by preincubation with calpain inhibitor I (N-acetyl-leucyl-leucyl-norleucinal [aLLN]), an active-site inhibitor. Both aLLN and PD150606, a specific calpain inhibitor that interacts with the calcium-binding site, inhibited reovirus-induced apoptosis in L929 cells by 54 to 93%. Apoptosis induced by UV-inactivated reovirus was also reduced 65 to 69% by aLLN, indicating that inhibition of apoptosis by calpain inhibitors is independent of effects on viral replication. We conclude that calpain activation is a component of the regulatory cascade in reovirus-induced apoptosis.


Reliability and validity of the body image quality of life inventory: version for Brazilian burn victims.
The aims of this study were to adapt the Body Image Quality of Life Inventory (BIQLI) into Brazilian Portuguese (BP) and to assess the psychometric properties of the adapted version. Construct validity was assessed by correlating the BIQLI-BP scores with the Rosenberg's Self-Esteem Scale, with Burns Specific Health Scale-Revised (BSHS-R), and with gender, total body surface area burned, and visibility of the scars. Participants were 77 adult burn patients. Cronbach's alpha for the adapted version was .90 and moderate linear correlations were found between body image and self-esteem and between BIQLI-BP scores and two domains of the BSHS-R: affect and body image and interpersonal relationships. The BIQLI-BP showed acceptable levels of reliability and validity for Brazilian burn patients.


Wnt signaling pathway in non-small cell lung cancer.
Wnt/β-catenin alterations are prominent in human malignancies. In non-small cell lung cancer (NSCLC), β-catenin and APC mutations are uncommon, but Wnt signaling is important in NSCLC cell lines, and Wnt inhibition reduces proliferation. Overexpression of Wnt-1, -2, -3, and -5a and of Wnt-pathway components Frizzled-8, Dishevelled, Porcupine, and TCF-4 is common in resected NSCLC and is associated with poor prognosis. Conversely, noncanonical Wnt-7a suppresses NSCLC development and is often downregulated. Although β-catenin is often expressed in NSCLCs, it was paradoxically associated with improved prognosis in some series, possibly because of E-cadherin interactions. Downregulation of Wnt inhibitors (eg, by hypermethylation) is common in NSCLC tumor cell lines and resected samples; may be associated with high stage, dedifferentiation, and poor prognosis; and has been reported for AXIN, sFRPs 1-5, WIF-1, Dkk-1, Dkk-3, HDPR1, RUNX3, APC, CDX2, DACT2, TMEM88, Chibby, NKD1, EMX2, ING4, and miR-487b. AXIN is also destabilized by tankyrases, and GSK3β may be inactivated through phosphorylation by EGFR. Preclinically, restoration of Wnt inhibitor function is associated with reduced Wnt signaling, decreased cell proliferation, and increased apoptosis. Wnt signaling may also augment resistance to cisplatin, docetaxel, and radiotherapy, and Wnt inhibitors may restore sensitivity. Overall, available data indicate that Wnt signaling substantially impacts NSCLC tumorigenesis, prognosis, and resistance to therapy, with loss of Wnt signaling inhibitors by promoter hypermethylation or other mechanisms appearing to be particularly important. Wnt pathway antagonists warrant exploration clinically in NSCLC. Agents blocking selected specific β-catenin interactions and approaches to increase expression of downregulated Wnt inhibitors may be of particular interest.


Actigraphy-measured sleep characteristics and risk of falls in older women.
Prior studies have suggested that insomnia and self-reported poor sleep are associated with increased risk of falls. However, no previous study, to our knowledge, has tested the independent associations of objectively estimated characteristics of sleep and risk of falls, accounting for the use of commonly prescribed treatments for insomnia. Study subjects were participants in the Study of Osteoporotic Fractures. In 2978 primarily community-dwelling women 70 years and older (mean age, 84 years), sleep and daytime inactivity were estimated using wrist actigraphy data collected for a minimum of 3 consecutive 24-hour periods (mean duration, 86.3 hours). Fall frequency during the subsequent year was ascertained by a triannual questionnaire. Use of medications was obtained by examiner interview. In multivariate-adjusted models, relative to those with "normal" nighttime sleep duration (>7 to 8 hours per night), the odds of having 2 or more falls in the subsequent year was elevated for women who slept 5 hours or less per night (odds ratio, 1.52; 95% confidence interval, 1.03-2.24). This association was not explained by the use of benzodiazepines. Indexes of sleep fragmentation were also associated with an increased risk of falls. For example, women with poor sleep efficiency (<70% of time in bed spent sleeping) had 1.36-fold increased odds of falling compared with others (odds ratio, 1.36; 95% confidence interval, 1.07-1.74). Short nighttime sleep duration and increased sleep fragmentation are associated with increased risk of falls in older women, independent of benzodiazepine use and other risk factors for falls.


[A device for balancing the cups in donor blood centrifugation].
It is important to well balance the loads of the opposite cups with plastic blood containers in order to correctly separate the donor blood into its components through centrifugation. A new balancer for centrifugation cups (USC5-01, manufacturer--Biophysical Instruments Ltd., Moscow) was tested at the Blood Center of Russia's Health Ministry. The original weights of all cups placed on the device weight sensors differed in all cases by readings of the light-emitting diode scale, which necessitated an appropriate balancing. The weight differences varied from 4 to 20 g, i.e. from 1 to 5 green lamps). The testing of the device verified its simplicity, convenience, and a shorter time spent on each balancing procedure; the device response lag to balancing made only 2-3 sec. The minimal out-of-balance index did not exceed 4 g, which ensures the obtaining of components meeting the requirements applied to them.


Lichen depsides and depsidones reduce symptoms of diseases caused by tobacco mosaic virus (TMV) in tobacco leaves.
Two depsides and five depsidones, isolated from lichens, were tested to determine their in vivo protective effects on tobacco leaves challenged with tobacco mosaic virus (TMV). The results indicate that most of these compounds are able to reduce either the number and/or the size of necrotic lesions following virus infection. Pannarin, 1'-chloro-pannarin and stictic acid provided the more effective protective results, reducing by at least 45% the number and size of lesions. Real Time PCR assays were used to explore the target of action against TMV by examining the response behavior of genes involved in the plant defense mechanism. The application of the lichen substances did not lead to changes in the transcriptional levels of pathogen-related (PR1a), allene oxide synthase 2 (AOS2) or oxophytodienoate reductase (OPR3) genes. Thus, the protection observed in the tobacco leaves treated with the lichen compounds may be mediated by a mechanism which does not involved the SA- or JA-mediated defensive plant response. A possible structure-activity relationship is presented.


Exploring the relationships between motor proficiency, independence and quality of life in adolescents with autism spectrum disorder.
The aim of this study was to explore the relationships between autism severity, motor performance, independence in daily living skills and quality of life in adolescents with autism spectrum disorder (ASD). The study included 31 participants, aged 13-18 years. Evaluations were made with the Gilliam Autistic Disorders Rating Scale-2 (GARS-2), Bruininks-Oseretsky Test of Motor Proficiency-2 Short Form (BOT-2-SF), Functional Independence Measure (WeeFIM II), and Pediatric Quality of Life Inventory (PedsQL). Motor performance of the participants was determined as below average according to the norms of BOT-2 SF, and were negatively correlated with GARS-2 at a moderate level (r = -0.61, p < 0.001), and a moderate negative correlation was determined between GARS-2 and WeeFIM II (r = -0.65, p < 0.001) and PedsQL (r = -0.61, p < 0.001). Independence in daily living skills (DLS) and autism severity were found to be significant predictors of PedsQL (R2 = 0.463, p < 0.05). Motor impairment still appears to be a missing piece of the puzzle for this age group. Promoting motor skill acquisition and independence could be a key factor in preventing the deterioration of physical health and improving quality of life in adolescents with ASD.


Lectin-tagged fluorescent polymeric nanoparticles for targeting of sialic acid on living cells.
In this study, we fabricated lectin-tagged fluorescent polymeric nanoparticles approximately 35 nm in diameter using biocompatible polymers conjugated with lectins for the purpose of detecting sialic acid on a living cell surface, which is one of the most important biomarkers for cancer diagnosis. Through cellular experiments, we successfully detected sialic acid overexpression on cancerous cells with high specificity. These fluorescent polymeric nanoparticles can be useful as a potential bioimaging probe for detecting diseased cells.


[A case of pancreatic cancer with local recurrence and liver metastases eight years after surgery].
Here we report a rare case of late recurrence of pancreatic cancer 8 years after surgery. A woman in her mid-fifties was hospitalized for examination of epigastralgia. Computed tomography (CT) revealed a 4 cm nodule at the pancreatic head with suspected invasion of the superior mesenteric vein. She underwent pancreaticoduodenectomy with wedge resection of superior mesenteric vein and intraoperative radiation therapy. Pathological findings showed moderately differentiated tubular adenocarcinoma and T3N1M0, Stage IIB according to The Union for International Cancer Control (UICC) TNM classification. As adjuvant chemotherapy, 56 courses of gemcitabine (GEM) were administered in 3.5 years. Because of long-term use of GEM, common terminology criteria for adverse events (CTCAE) Grade 3 anemia occurred, and chemotherapy was discontinued. Tumor markers were evaluated every month and CT scans were taken every 6 months for 5 years. Subsequently, CT was performed annually. The patient was hospitalized for high-grade fever, 8.5 years after surgery. CT, magnetic resonance imaging (MRI) and positron emission tomography-computed tomography (PET-CT) detected local recurrence with liver metastases. GEM was administered again, but was ineffective. The patient died 9 years after surgery. In conclusion, even if long-term survival is achieved in pancreatic cancer, follow-ups should not be stopped.


Characterizing Spatial Information Loss for Wastewater Surveillance Using crAssphage: Effect of Decay, Temperature, and Population Mobility.
Populations contribute information about their health status to wastewater. Characterizing how that information degrades in transit to wastewater sampling locations (e.g., wastewater treatment plants and pumping stations) is critical to interpret wastewater responses. In this work, we statistically estimate the loss of information about fecal contributions to wastewater from spatially distributed populations at the census block group resolution. This was accomplished with a hydrologically and hydraulically influenced spatial statistical approach applied to crAssphage (Carjivirus communis) load measured from the influent of four wastewater treatment plants in Hamilton County, Ohio. We find that we would expect to observe a 90% loss of information about fecal contributions from a given census block group over a travel time of 10.3 h. This work demonstrates that a challenge to interpreting wastewater responses (e.g., during wastewater surveillance) is distinguishing between a distal but large cluster of contributions and a near but small contribution. This work demonstrates new modeling approaches to improve measurement interpretation depending on sewer network and wastewater characteristics (e.g., geospatial layout, temperature variability, population distribution, and mobility). This modeling can be integrated into standard wastewater surveillance methods and help to optimize sewer sampling locations to ensure that different populations (e.g., vulnerable and susceptible) are appropriately represented.


Changes in the lymphocyte response and in the production of IgE regulatory factors in relation to the course of immunotherapy.
In previous studies we described isotype-specific but antigen non-restricted soluble factors produced by human lymphocytes. In subsequent studies we demonstrated changes in the production of these factors during the course of immunotherapy (IT) by testing them on lymphocytes from normal healthy controls and allergic patients. Through these studies we confirmed that there exists a difference in lymphocytes' responsiveness to soluble factors between both groups. In this report, we investigated the effect of soluble factors on lymphocytes from allergic patients without IT (LyG1) and with IT longer than 2 years (LyG4). Peripheral blood samples were collected from healthy controls and allergic patients at different time periods of IT, and bidirectional mixed cultures were performed with the isolated lymphocytes. Supernatants obtained from chromatography were tested on lymphocytes of allergic patients without IT and with IT greater than 2 years to determine their effect on IgE synthesis. Long periods of IT reduce the production of Suppressor Factors (SF) by allergic patients as well as their responsiveness. Long periods of IT increase the responsiveness of lymphocytes of allergic patients to Enhancing Factors (EF) and decrease EF production. We propose a "receptor hypothesis" to explain these events.


A "HOWFARLESS" option to increase efficiency of homogeneous phantom calculations with DOSXYZnrc.
This paper describes a "HOWFARLESS" transport option, which has been added to DOSXYZnrc to increase the efficiency of beam commissioning calculations in homogeneous phantoms. The algorithm speeds up charged particle transport by only considering the distance to the extreme outer boundaries of the phantom, thus eliminating the need to stop at voxel boundaries. Dose is deposited by approximating the total curved charged particle steps by two straight-line steps joined at a hinge point. Good agreement with normal simulations is achieved at all beam energies and for all practical maximum step lengths with a 1:1 mixture of approximations based on the initial position/ direction of the particle and on its final position/direction. Use of the "HOWFARLESS" option in phantom calculations for 6 and 18 MV photon beams (10 x 10 cm2 and 40 x 40 cm2 fields) from BEAMnrc-simulated accelerators increases the efficiency at the optimum photon splitting number by a factor of 2.9-5.4 when the exact EGSnrc boundary crossing algorithm (BCA) is used and by 51%-89% when the faster PRESTA-I BCA is employed. The efficiency gain due to the "HOWFARLESS" transport option increases with increasing beam energy and decreases with increasing field/dose voxel size. Efficiency improvement is greater when the efficiency of the particle source itself is not a factor, and in such cases the "HOWFARLESS" option improves the DOSXYZnrc efficiency by up to a factor of 13.1 (exact BCA) or 3.5 (PRESTA-I BCA) for photon beams, and up to a factor of 17.2 (exact BCA) or 5.2 (PRESTA-I BCA) for electron beams.


Revealing cell assemblies at multiple levels of granularity.
Current neuronal monitoring techniques, such as calcium imaging and multi-electrode arrays, enable recordings of spiking activity from hundreds of neurons simultaneously. Of primary importance in systems neuroscience is the identification of cell assemblies: groups of neurons that cooperate in some form within the recorded population. We introduce a simple, integrated framework for the detection of cell-assemblies from spiking data without a priori assumptions about the size or number of groups present. We define a biophysically-inspired measure to extract a directed functional connectivity matrix between both excitatory and inhibitory neurons based on their spiking history. The resulting network representation is analyzed using the Markov Stability framework, a graph theoretical method for community detection across scales, to reveal groups of neurons that are significantly related in the recorded time-series at different levels of granularity. Using synthetic spike-trains, including simulated data from leaky-integrate-and-fire networks, our method is able to identify important patterns in the data such as hierarchical structure that are missed by other standard methods. We further apply the method to experimental data from retinal ganglion cells of mouse and salamander, in which we identify cell-groups that correspond to known functional types, and to hippocampal recordings from rats exploring a linear track, where we detect place cells with high fidelity. We present a versatile method to detect neural assemblies in spiking data applicable across a spectrum of relevant scales that contributes to understanding spatio-temporal information gathered from systems neuroscience experiments.


Asparagine 175 of connexin32 is a critical residue for docking and forming functional heterotypic gap junction channels with connexin26.
The gap junction channel is formed by proper docking of two hemichannels. Depending on the connexin(s) in the hemichannels, homotypic and heterotypic gap junction channels can be formed. Previous studies suggest that the extracellular loop 2 (E2) is an important molecular domain for heterotypic compatibility. Based on the crystal structure of the Cx26 gap junction channel and homology models of heterotypic channels, we analyzed docking selectivity for several hemichannel pairs and found that the hydrogen bonds between E2 domains are conserved in a group of heterotypically compatible hemichannels, including Cx26 and Cx32 hemichannels. According to our model analysis, Cx32N175Y mutant destroys three hydrogen bonds in the E2-E2 interactions due to steric hindrance at the heterotypic docking interface, which makes it unlikely to dock with the Cx26 hemichannel properly. Our experimental data showed that Cx26-red fluorescent protein (RFP) and Cx32-GFP were able to traffic to cell-cell interfaces forming gap junction plaques and functional channels in transfected HeLa/N2A cells. However, Cx32N175Y-GFP exhibited mostly intracellular distribution and was occasionally observed in cell-cell junctions. Double patch clamp analysis demonstrated that Cx32N175Y did not form functional homotypic channels, and dye uptake assay indicated that Cx32N175Y could form hemichannels on the cell surface similar to wild-type Cx32. When Cx32N175Y-GFP- and Cx26-RFP-transfected cells were co-cultured, no colocalization was found at the cell-cell junctions between Cx32N175Y-GFP- and Cx26-RFP-expressing cells; also, no functional Cx32N175Y-GFP/Cx26-RFP heterotypic channels were identified. Both our modeling and experimental data suggest that Asn(175) of Cx32 is a critical residue for heterotypic docking and functional gap junction channel formation between the Cx32 and Cx26 hemichannels.


The advanced glycation end product pentosidine correlates to IL-6 and other relevant inflammatory markers in rheumatoid arthritis.
Oxidative stress and inflammatory processes accelerate the formation of advanced glycation end products (AGE), e.g. of pentosidine. The aim of this study was to investigate the relationships between levels of pentosidine in serum and synovial fluid, proinflammatory cytokines, other markers of inflammatory activity, and the state of radiologically visible bone destruction in patients with rheumatoid arthritis (RA). One hundred thirty-three nondiabetic RA patients and 56 age-matched, healthy subjects were included. Serum and synovial fluid pentosidine, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and rheumatoid factor levels were determined. In 30 patients, the proinflammatory cytokines interleukin (IL)-1beta, IL-6, and TNF-alpha and the soluble receptors sIL-2R, sIL-6R, sTNF-alpha, and RI/RII were also measured. Serum levels of pentosidine were on average significantly higher in RA patients than in healthy subjects and correlated significantly to ESR, CRP, and serum levels of IL-6. Serum and synovial fluid pentosidine did not show any differences. Rheumatoid factor-positive RA patients had higher pentosidine levels in the synovial fluid than rheumatoid factor-negative patients. Correlations could not be found between pentosidine and the other cytokines or cytokine receptors measured. The binding of AGE on cell receptors induces activation of nuclear factor kappa B, resulting in enhanced synthesis of proinflammatory cytokines. Moreover, AGE generation may also lead to the formation of new, immunologically relevant epitopes at synovial proteins. Both mechanisms could contribute to initiation and perpetuation of the inflammatory and destructive processes in RA.


Specific T-cell responses for guiding treatment with convalescent plasma in severe COVID-19 and humoral immunodeficiency: a case report.
The immune response to SARS-CoV-2 virus, the cause of COVID-19, is complex. Antibody mediated responses are important for viral clearance but may also drive hyperinflammation in severe COVID-19. We present a case of an individual with a genetic inability to produce antibodies and severe COVID-19, receiving no other specific anti-viral treatment than convalescent COVID-19 plasma, illustrating that hyperinflammation can occur in the absence of a humoral anti-viral response. In addition, the case illustrates that the assessment of SARS-CoV-2 T cell responses can facilitate clinical decision making in patients with COVID-19 and weak or absent humoral immune responses. A male with X-linked agammaglobulinemia on regular immunoglobulin replacement therapy, hospitalized for 35 days due to severe COVID-19. Systemic inflammatory parameters were highly elevated. After treatment with convalescent COVID-19 plasma he became afebrile and the fatigue diminished. He was discharged on day 42 and nasopharyngeal SARS-CoV-2 PCR eventually was negative on day 49. Evidence of SARS-CoV-2 specific T cells prior to administration of plasma therapy suggested that antibodies were crucial for viral clearance. Regular assessment showed robust and persistent SARS-CoV-2 specific T-cell responses after recovery suggested that prophylactic administration of convalescent COVID-19 plasma was unnecessary. Assessment of SARS-CoV-2T-cell responses can facilitate the clinical management of COVID-19 patients with humoral immunodeficiencies.


Low-cost and wideband frequency tunable optoelectronic oscillator based on a directly modulated distributed feedback semiconductor laser.
A novel scheme to realize a low-cost and wideband frequency tunable optoelectronic oscillator based on a directly modulated distributed feedback (DFB) semiconductor laser is proposed and experimentally demonstrated. In the proposed scheme, neither an external modulator nor an electrical filter is used, and no more than 25 dB of the electrical loop gain is required due to the high modulation efficiency of the relaxation oscillation frequency of the DFB laser. Microwave signals with frequency coarsely tuned from 3.77 to 8.75 GHz are generated by changing the bias current and operation temperature of the DFB laser. The single sideband phase noise of the generated 6.97 GHz microwave signal is measured to be -103.6 dBc/Hz at 10 kHz offset.


DNA photolyases of Chrysodeixis chalcites nucleopolyhedrovirus are targeted to the nucleus and interact with chromosomes and mitotic spindle structures.
Cyclobutane pyrimidine dimer (CPD) photolyases convert UV-induced CPDs in DNA into monomers using visible light as the energy source. Two phr genes encoding class II CPD photolyases PHR1 and PHR2 have been identified in Chrysodeixis chalcites nucleopolyhedrovirus (ChchNPV). Transient expression assays in insect cells showed that PHR1-EGFP fusion protein was localized in the nucleus. Early after transfection, PHR2-EGFP was distributed over the cytoplasm and nucleus but, over time, it became localized predominantly in the nucleus. Immunofluorescence analysis with anti-PHR2 antiserum showed that, early after transfection, non-fused PHR2 was already present mainly in the nucleus, suggesting that the fusion of PHR2 to EGFP hindered its nuclear import. Both PHR-EGFP fusion proteins strongly colocalized with chromosomes and spindle, aster and midbody structures during host-cell mitosis. When PHR2-EGFP-transfected cells were superinfected with Autographa californica multiple-nucleocapsid NPV (AcMNPV), the protein colocalized with virogenic stroma, the replication factories of baculovirus DNA. The collective data support the supposition that the PHR2 protein plays a role in baculovirus DNA repair.


Diminished IFN-gamma response to diabetes-associated autoantigens in children at diagnosis and during follow up of type 1 diabetes.
Imbalance of T-helper (Th)1- and Th2-like cytokines has been associated with type 1 diabetes. We therefore studied the immune deviation in antigen-specific T cells from diagnosis onwards in type 1 diabetic children. Peripheral blood mononuclear cells (PBMC) were collected from 15 children after 4 days, 3 months and 18 months of being diagnosed with type 1 diabetes, from 15 healthy children matched by age and gender to the type 1 diabetic children and from 14 children with and 35 children without HLA-risk genes. Secretion of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) was detected by ELISPOT after stimulation with glutamic acid decarboxylase (GAD(65), protein and aa 247-279), recombinant tyrosinphosphatase (IA-2), insulin, ovalbumin and phytohaemagglutinin (PHA). Secretion of IFN-gamma in PBMC stimulated with GAD(65) (p < 0.05), the GAD(65)-peptide (p < 0.01), IA-2 (p < 0.01), and insulin (p < 0.01) was lower in diabetic children at diagnosis than in healthy children. Stimulation of PBMC with GAD(65) and IA-2 decreased the secretion of IFN-gamma in children with HLA-risk genotype. Spontaneous and antigen-induced IFN-gamma secretion increased significantly after diagnosis of the disease, but did not exceed the levels observed in healthy children. Fasting C-peptide levels at diagnosis correlated with insulin-induced IFN-gamma (R = 0.52; p = 0.05) and negatively with spontaneous IL-4 secretion (R = -0.62; p < 0.05). A diminished IFN-gamma secretion and the association of fasting C-peptide levels with cytokine response in children with type 1 diabetes suggest that factors related to beta-cell function in type 1 diabetes may modify T-cell function. Thus, the T-cell responses detected at or after diagnosis may not reflect the pathogenic process leading to type 1 diabetes.


Inactivation of gamma-glutamylcysteine synthetase by SAPH-3 disulfide: role of the histaminyl moiety.
Certain disulfide analogues of cystamine were prepared and evaluated for the ability to inhibit gamma-glutamylcysteine synthetase (GCS), the rate-limiting enzyme in glutathione synthesis. The compound SAPH-3 disulfide (2a) was found to be the most effective inactivator of GCS reported to date. Studies of structural analogues of 2a indicate that the histamine moiety plays a significant role in enzyme inactivation. Substitution of methyl groups on the carbon atoms adjacent to both sulfur atoms of the disulfide drastically decreases inhibitory action, probably due to a steric repulsion near the sulfur atom at the enzyme active site.