Biomechanical study of a unilocking T-plate system for prophylactic internal fixation of the radial osteocutaneous donor site using the sheep tibia model.
Prophylactic internal fixation (PIF), with a bone plate in either the anterior (over the section defect) or posterior (on intact cortex) position, has substantially reduced the incidence of fracture at the donor site of the radial osteocutaneous free flap. This study uses the sheep tibia model to compare the effectiveness of new T-shaped titanium plates utilising a unilocking screw system with a 3.5 mm steel plate and bicortical screw fixation system commonly applied for PIF. Forty matched pairs of adult sheep tibias were tested in torsion and 4-point bending. An osteotomised bone was significantly weaker (p<0.001) than an intact bone in both bending and torsion with a mean loss of 77% and 64% of strength respectively. The tibia withstood much greater bending loads. All of the constructs significantly strengthened an osteotomised bone by a factor of 1.73-2.43 times in bending and 1.54-2.63 in torsion. The 2.4 mm T-plate in an anterior position (section) was the baseline against which other plates in differing positions were compared. The 3.5 mm T-plate section, DCP section and DCP cortex constructs had 41%, 30% and 2% greater mean bending strengths respectively but only the 3.5 mm T-plate section result approached statistical significance (p=0.06). In torsion the DCP section, 3.5 mm T-plate section and DCP cortex constructs had 56% (p=0.01), 27% (p=0.06) and 25% greater mean strengths respectively. When compared to an intact bone the mean bending strength restored by the DCP section (84%) and 3.5 mm T-plate section (87%) constructs was greatest and effectively restored the strength to that of an intact bone (100%). In torsion the mean strength restored by the DCP section (62%), DCP cortex (44%), 3.5 mm T-plate section (40%) and 2.4 mm T-plate (36%) remained significantly less than an intact bone. All of the plate constructs significantly strengthened an osteotomised bone but overall the 3.5 mm T-plate section and DCP section were the strongest constructs and most suitable for PIF. The lighter 2.4 mm T-shaped titanium plate was least effective. The strongest reinforcement in bending and torsion was the 3.5 mm T-plate section and DCP section respectively. The 3.5 mm DCP section plate was significantly stronger (p=0.01) than the 3.5 mm T-plate in torsion and remains the most effective construct for resisting torsional stresses, which are probably the commonest cause of fracture in clinical practice.


Bacteriophage transfer during faecal microbiota transplantation in Clostridium difficile infection is associated with treatment outcome.
Faecal microbiota transplantation (FMT) is effective for the treatment of recurrent Clostridium difficile infection (CDI). Studies have shown bacterial colonisation after FMT, but data on viral alterations in CDI are scarce. We investigated enteric virome alterations in CDI and the association between viral transfer and clinical outcome in patients with CDI. Ultra-deep metagenomic sequencing of virus-like particle preparations and bacterial 16S rRNA sequencing were performed on stool samples from 24 subjects with CDI and 20 healthy controls. We longitudinally assessed the virome and bacterial microbiome changes in nine CDI subjects treated with FMT and five treated with vancomycin. Enteric virome alterations were assessed in association with treatment response. Subjects with CDI demonstrated a significantly higher abundance of bacteriophage Caudovirales and a lower Caudovirales diversity, richness and evenness compared with healthy household controls. Significant correlations were observed between bacterial families Proteobacteria, Actinobacteria and Caudovirales taxa in CDI. FMT treatment resulted in a significant decrease in the abundance of Caudovirales in CDI. Cure after FMT was observed when donor-derived Caudovirales contigs occupied a larger fraction of the enteric virome in the recipients (p=0.024). In treatment responders, FMT was associated with alterations in the virome and the bacterial microbiome, while vancomycin treatment led to alterations in the bacterial community alone. In a preliminary study, CDI is characterised by enteric virome dysbiosis. Treatment response in FMT was associated with a high colonisation level of donor-derived Caudovirales taxa in the recipient. Caudovirales bacteriophages may play a role in the efficacy of FMT in CDI. NCT02570477.


Menopause: plight or passage?
Although the menopause has been discussed in medical, biological, and social science literature, the experience of menopause for women has been shrouded in myths and taboos. Separating myth from reality has been difficult for the American woman, who traditionally has not sought medical advice about the menopause on a routine basis. Health care professionals need to understand the influence of history, culture, taboos, and myths on the symptomatology and interpretation of the menopausal experience.


Research or quality improvement?. Making the decision.
Deciding if a project is one of research or quality improvement is a dilemma frequently faced by nursing administrators. Guidelines have been established to help administrators and practitioners overcome this dilemma and, at the same time, consider the rights and responsibilities of the patient, the hospital, and the investigator.


Colonization and diversification of the species Brachyderes rugatus (Coleoptera) on the Canary Islands: evidence from mitochondrial DNA COII gene sequences.
The genus Brachyderes Schönherr (Coleoptera: Curculionidae) is represented by the species B. rugatus Wollaston on the Canary Islands, with one subspecies on each of the islands of Gran Canaria, Tenerife, La Palma, and El Hierro. These four subspecies are associated with the endemic pine tree Pinus canariensis, and their distributions are broadly coincident. Eighty-eight individual Canarian Brachyderes, sampled from across the distributions of each subspecies, have been sequenced for 570 bp of the mitochondrial DNA (mtDNA) cytochrome oxidase II gene (COII). No mitotypes are shared among islands. Both maximum-likelihood and distance-based phylogenetic analyses have shown that: Tenerife is composed of a single monophyletic clade of mitotypes, El Hierro is composed of a single monophyletic clade occurring within a larger clade comprising all the La Palma mitotypes, and the mitotypes of these three islands form a monophyletic group distinct from Gran Canaria. New methods for estimating divergence times without the assumption of rate constancy have been used to reconstruct the direction and approximate timing of colonizations among the islands. Colonization has occurred from older to progressionally younger islands, and these colonizations are estimated to have occurred less than 2.6 million years ago, although the timing of the initial colonization of the archipelago is not discernable. New methods for the estimation of diversification rates that use branching times as the analyzed variable have been applied to each island fauna. Hypothesized effects of different levels of recent volcanism among islands were not apparent. All islands exhibit a gradually decreasing rate of genetic diversification that is marked by periodic sudden changes in rate.


Intracellular Staphylococcus aureus escapes the endosome and induces apoptosis in epithelial cells.
We examined the invasion of an established bovine mammary epithelial cell line (MAC-T) by a Staphylococcus aureus mastitis isolate to study the potential role of intracellular survival in the persistence of staphylococcal infections. S. aureus cells displayed dose-dependent invasion of MAC-T cells and intracellular survival. An electron microscopic examination of infected cells indicated that the bacteria induced internalization via a mechanism involving membrane pseudopod formation and then escaped into the cytoplasm following lysis of the endosomal membrane. Two hours after the internalization of S. aureus, MAC-T cells exhibited detachment from the matrix, rounding, a mottled cell membrane, and vacuolization of the cytoplasm, all of which are indicative of cells undergoing programmed cell death (apoptosis). By 18 h, the majority of the MAC-T cell population exhibited an apoptotic morphology. Other evidence for apoptosis was the generation of MAC-T cell DNA fragments differing in size by increments of approximately 180 bp and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling of the fragmented nuclear DNA of the infected host cells. These results demonstrate that after internalization S. aureus escapes the endosome and induces apoptosis in nonprofessional phagocytes.


Seasonal changes in wood formation of Ulmus pumila and U. minor and its relation with Dutch elm disease.
Elms containing narrow and scattered vessels have been reported to be more resistant to Ophiostoma novo-ulmi (Dutch elm disease pathogen) than elms with large and contiguous vessels. However, recent measurements in Ulmus pumila and U. minor showed a contrary trend. The pin method was applied to 4-yr-old branches of eight clones planted in Madrid. During 2002, radial growth increments and vessel diameters were measured monthly, and beetle trapping was undertaken weekly. U. minor formed larger vessels at the beginning of the season, coinciding with a peak of captured beetles, but, up to June 15, vessels were larger for U. pumila. The number of vessels per group, the transversal area per vessel group, and the mean theoretical hydraulic conductances were significantly higher for U. minor on most dates. Researchers should take into consideration the seasonal changes in vessel size. The results highlight that seasonal variation of vessel diameters and hydraulic parameters, in combination with beetle abundance, are the main factors that could explain the different susceptibility of both elm species to O. novo-ulmi.


[Mechanism of action of gliquidone (author's transl)].
A study of 13 metabolically normal persons, 12 patients with asymptomatic or latent diabetes and 11 with manifest diabetes showed that 30 mg gliquidone orally prevents or delays the blood sugar rise during repeated small carbohydrate meals with simultaneous stimulation of insulin secretion. This effect was less marked in the normal and asymptomatic or latent diabetic subjects than after glibenclamide 5 mg, while there was no difference between the two drugs in those with manifest diabetes.


Structure and dynamics of an Arp2/3 complex-independent component of the lamellipodial actin network.
Sea urchin coelomocytes contain an unusually broad lamellipodial region and have served as a useful model experimental system for studying the process of actin-based retrograde/centripetal flow. In the current study the small molecule drug 2,3-butanedione monoxime (BDM) was employed as a means of delocalizing the Arp2/3 complex from the cell edge in an effort to investigate the Arp2/3 complex-independent aspects of retrograde flow. Digitally-enhanced phase contrast, fluorescence and polarization light microscopy, along with rotary shadow transmission electron microscopy methods demonstrated that BDM treatment resulted in the centripetal displacement of the Arp2/3 complex and the associated dendritic lamellipodial (LP) actin network from the cell edge. In its wake there remained an array of elongate actin filaments organized into concave arcs that displayed retrograde flow at approximately one quarter the normal rate. Actin polymerization inhibitor experiments indicated that these arcs were generated by polymerization at the cell edge, while active myosin-based contraction in BDM treated cells was demonstrated by localization with antiphospho-myosin regulatory light chain (MRLC) antibody, the retraction of the cytoskeleton in the presence of BDM, and the response of the BDM arcs to laser-based severing. The results suggest that BDM treatment reveals an Arp2/3 complex-independent actin structure in coelomocytes consisting of elongate filaments integrated into the LP network and that these filaments represent a potential connection between the LP network and the central cytoskeleton.


The PDZ protein discs-large (DLG): the 'Jekyll and Hyde' of the epithelial polarity proteins.
Discs-large (DLG) is a multi-PDZ domain-containing protein that belongs to the family of molecular scaffolding proteins known as membrane guanylate kinases or MAGUKs. DLG is a component of the Scribble polarity complex and genetic analyses of DLG in Drosophila have identified a role for the protein in several key biological processes including the regulation of apico-basal polarity of epithelial cells, as well as other polarity processes such as asymmetric cell division and cell invasion. Disturbance of DLG function leads to uncontrolled epithelial cell proliferation and neoplastic transformation, thereby defining DLG as a potential tumour suppressor. However, whether mammalian homologues of DLG (DLG1, DLG2, DLG3 and DLG4) also possess tumour suppressor functions is not known. In this minireview, we focus on the biological functions of DLG1 in human epithelial cells and on how the function of this MAGUK relates to its intracellular location. We examine some of the evidence that implies that DLG has both tumour suppressor and, paradoxically, oncogenic functions depending upon the precise cellular context.


Emodin attenuates adenosine triphosphate‑induced pancreatic ductal cell injury in vitro via the inhibition of the P2X7/NLRP3 signaling pathway.
Acute pancreatitis (AP) is an inflammatory disease with high morbidity and mortality rates. Pancreatic ductal cells are the most susceptible of all cell types that are exposed to the noxious stimuli of pancreatitis. Our previous studies demonstrated that emodin, a natural product extracted from Rheum palmatum L., protected pancreatic acinar cells from injutyh due to its anti‑inflammatory activity. In the present study, in order to investigate the protective effects and molecular mechanisms of action of emodin on injured pancreatic ductal cells, an adenosine triphosphate (ATP)‑induced model of cell injury was established using the human pancreatic ductal epithelial cell line, HPDE6‑C7. The results revealed that emodin attenuated ATP‑induced HPDE6‑C7 cell injury by decreasing the levels of inflammatory factors, including interleukin (IL)‑1β and IL‑18. Furthermore, emodin significantly downregulated the protein levels of purinergic receptor P2X, ligand‑gated ion channel, 7 (P2X7), NOD‑like receptor protein 3 (NLRP3), apoptosis‑associated speck‑like protein containing a CARD (ASC) and caspase‑1 in the injured HPDE6‑C7 cells. The results also indicated that emodin attenuated HPDE6‑C7 cell injury at least partially through the inhibition of the P2X7/NLRP3 signaling pathway. The protective effects of emodin were abrogated upon pre‑treatment with P2X7 overexpression plasmid, which further confirmed that the P2X7 signaling pathway is the drug target of the effects of emodin against ATP‑induced pancreatic ductal cell injury. Collectively, the findings of this study demonstrate that emodin attenuates ATP‑induced pancreatic ductal cell injury in AP mainly through the inhibition of the P2X7/NLRP3 signaling pathway. This study suggests that emodin may be further developed for its application in future medical therapy.


Impact of naled on the mosquito vectors of eastern equine encephalitis virus.
This letter questions the appropriateness of methodology used in a study by Howard and Oliver (J. Am. Mosq. Control Assoc. 13:315-325; 1988). Two independent data sets, collected for different purposes by 2 different groups, were subjected to statistical analysis to determine if the data sets differed. The experimental "design," as described by the authors, is an example of pseudoreplication, which arises when replicates are collected at a scale finer than the one for which conclusions of statistical testing are intended to be drawn. All of the components of a properly designed field experiment (control, replication, randomization, and interspersion) are missing from this study. The authors proceed to draw a series of conclusions from the data presented. Few, if any, of the conclusions can be supported by the evidence presented. The assertions put forward in this paper could have a severe negative impact on efforts to prevent transmission of arboviruses or other pathogens to humans and domestic animals.


[Hormonal treatment of undescended testis].
Failure of the testes to descend is probably mostly due to dysfunction of the hypothalamo-pituitary-testis axis. The primary treatment of testicular non descent should therefore be pharmacological in the form of HCG injections or LH-RH intranasally. Contraindications for hormonal therapy are coexistent inguinal hernia or hydrocele testis or previous inguinal operations. HCG and LH-RH treatment lead to testicular descent with about the same frequency, 20-50% in boys above five years of age, whereas HCG therapy before this age results in 10-20% success only. Reversible androgenic side effects are described in approximately 30% following HCG treatment, but only limited side effects are seen in less than 10% after LH-RH therapy. Because of the non-invasive mode of administration, the good response to treatment in the age of 2-5 years and the very limited side effects, LH-RH is recommended as the primary treatment of testicular nondescent.


Therapeutic monitoring of carboplatin dosing in a premature infant with retinoblastoma.
Carboplatin dosing based on renal function and therapeutic monitoring have been previously shown to be beneficial in the treatment of children with cancer. However, the applicability of such approaches to the treatment of premature or newborn infants, where kidney function may change markedly with advancing gestational and postnatal age, is unknown. Diagnosis of retinoblastoma in a preterm infant provided a rare opportunity to carry out adaptive carboplatin dosing in a patient with immature renal function. A preterm female infant born at a gestational age of 32 weeks was diagnosed with bilateral retinoblastoma at 35 weeks. Carboplatin treatment with real-time pharmacokinetic monitoring was initiated on day 26 of life at an initial dose of 6.6 mg/kg. Plasma samples were obtained at specified time points and carboplatin levels quantified by atomic absorption spectrometry. Additional doses of carboplatin were determined by pharmacokinetic monitoring based on the achievement of carboplatin AUC values of 5.2-7.8 mg/ml min on three courses of treatment. Increased carboplatin doses administered on successive courses of treatment reflected a greater than twofold increase in drug clearance, from 3.4-7.1 ml/min over a 7-week period. Pharmacokinetically-guided carboplatin dosing led to the attainment of AUCs within 10% of target values on each course of treatment. The patient completed five courses of carboplatin with both tumours defined as inactive after this treatment period. Data obtained from studying this patient suggests that adaptive carboplatin monitoring represents a feasible and beneficial clinical approach in preterm infants or neonates.


Seeing What You Believe or Believing What You See? Belief Biases Correlation Estimation.
When an analyst or scientist has a belief about how the world works, their thinking can be biased in favor of that belief. Therefore, one bedrock principle of science is to minimize that bias by testing the predictions of one's belief against objective data. But interpreting visualized data is a complex perceptual and cognitive process. Through two crowdsourced experiments, we demonstrate that supposedly objective assessments of the strength of a correlational relationship can be influenced by how strongly a viewer believes in the existence of that relationship. Participants viewed scatterplots depicting a relationship between meaningful variable pairs (e.g., number of environmental regulations and air quality) and estimated their correlations. They also estimated the correlation of the same scatterplots labeled instead with generic 'X' and 'Y' axes. In a separate section, they also reported how strongly they believed there to be a correlation between the meaningful variable pairs. Participants estimated correlations more accurately when they viewed scatterplots labeled with generic axes compared to scatterplots labeled with meaningful variable pairs. Furthermore, when viewers believed that two variables should have a strong relationship, they overestimated correlations between those variables by an r-value of about 0.1. When they believed that the variables should be unrelated, they underestimated the correlations by an r-value of about 0.1. While data visualizations are typically thought to present objective truths to the viewer, these results suggest that existing personal beliefs can bias even objective statistical values people extract from data.


Effects of immediate performance feedback on implementation of behavior support plans.
Research has focused on increasing the treatment integrity of school-based interventions by utilizing performance feedback. The purpose of this study was to extend this literature by increasing special education teachers' treatment integrity for implementing antecedent and consequence procedures in an ongoing behavior support plan. A multiple baseline across teacher-student dyads (for two classrooms) design was used to evaluate the effects of performance feedback on the percentage of antecedent and consequence components implemented correctly during 1-hr observation sessions. Performance feedback was provided every other week for 8 to 22 weeks after a stable or decreasing trend in the percentage of antecedent or consequence components implemented correctly. Results suggested that performance feedback increased the treatment integrity of antecedent components for 4 of 5 teachers and consequence components for all 5 teachers. These results were maintained following feedback for all teachers across antecedent and consequence components. Teachers rated performance feedback favorably with respect to the purpose, procedures, and outcome, as indicated by a social validity rating measure.


Meckel's diverticulum masked by a long period of intermittent recurrent subocclusive episodes.
Meckel's diverticulum (MD) is the most frequent congenital abnormality of the small bowel and it is often difficult to diagnose. It is usually asymptomatic but approximately 4% are symptomatic with complications such as bleeding, intestinal obstruction, and inflammation. The authors report a case of a 7-year-old boy with a one-year history of recurrent periumbilical colicky pain with associated alimentary vomiting, symptoms erroneously related to a cyclic vomiting syndrome but not to MD. The clinical features and the differential diagnostic methods employed for diagnosis of MD are discussed.


A racial comparison of combat veterans evaluated for PTSD.
This study attempted to replicate the work of Frueh, Smith, and Libet (1996), which showed racial differences on psychological measures of dissociation/thought disturbance and the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) F-K index in combat veterans evaluated for posttraumatic stress disorder (PTSD). Veterans completed the Beck Depression Inventory, Mississippi Scale for Combat-Related PTSD, a fixed-response format version of the Dissociative Experiences Scale (DES-FRF), and MMPI-2 prior to treatment at a Veterans Affairs hospital outpatient PTSD clinic. Contrary to expectation, significant racial differences on the DES-FRF, MMPI-2 validity scales, and MMPI-2 Scales 6 and 8 were not found. Consistent with the previous study, no racial differences on measures of anxiety, depression, or PTSD symptomatology were found; nor were there racial differences on clinician ratings of global assessment of functioning or on most categories of psychiatric diagnoses. This suggests that Black and White combat veterans evaluated for PTSD do not differ with regard to reported manifestation or severity of psychopathology.


The Effect of Mofettes on Oxidative Stress/Antioxidant Balance in Experimental Myocardial Ischemia.
Natural mofettes are gases resulting from post-volcanic emanations. This study aimed to examine the effect of mofette therapy on plasma oxidative stress and antioxidant parameters in rats after experimental induction of myocardial ischemia, as well as on structural changes in myocardial tissue. White Wistar-Bratislava rats were divided into three groups. In groups 2 and 3, myocardial ischemia was induced by isoproterenol. Rats in group 3 were additionally exposed to high levels mofettes. Oxidative stress and antioxidant parameters were determined in plasma. The structural changes of the myocardium were observed in paraffin embedded slices contrasted using Goldner's trichrome staining. A statistically significant change in serum oxidative stress biomarkers, including nitric oxide, malondialdehyde, total oxidant status, as well as in the tested antioxidant molecules and total antioxidant capacity were observed in group 3 compared to group 2. Also, rats of group 3 showed an obvious improvement in inflammatory infiltration and repair of necrotic areas through collagen proliferation (proliferation of fibrous connective tissue) compared to group 2. Mofette had a beneficial effect on the balance between oxidative stress and antioxidant status following experimentally induced myocardial ischemia.


Breast cancer: postoperative radiation therapy.
Postoperative radiation therapy reduces the proportion of regional recurrences following radical mastectomy for cancer of the breast. Only a small number of patients with tumor beyond surgical effectiveness and without distant metastases stand to benefit. Improvement of final results due to postoperative radiation therapy can only be shown in large series of cases.


[Biologically active substances isolated from Hypericum attenuatum Choisy].
Four dianthrones, two oxymethylanthraquinones, two reductive forms of anthraquinones and five flavonoids have been isolated from the plant source Hypericum attenuatum Choisy (Guttiferae) firstly investigated for these substances contents. Dianthrones pseudohypericin, protohypericin, protopseudohypericin, oxymethylanthraquinones frangulaemodin, frangulin, reductive anthraquinones emodin anthrone, emodin anthranol, and flavonoids quercetin, quercitrin, isoquercitrin, rutin have been identified for the first time. High hyperoside contents in herb of Hypericum attenuatum is established.


Evaluation of feasibility of ankle pressure and foot oximetry values for the detection of critical limb ischemia in diabetic patients.
A total of 261 diabetic patients were admitted because of rest pain and/or foot ulcer in 1 limb. Ankle pressure (AP) and transcutaneous oxygen tension (TcPO(2)) were measured, and digital subtraction arteriography was performed. Transcutaneous oxygen tension was <30 mm Hg in 213 patients and >or=30<50 mm Hg in 48 patients. Ankle pressure could not be measured in 109 patients. In 50 patients, AP was <70 mm Hg and in 102 patients, it was >or=70 mm Hg. Arteriography showed evidence of stenoses >50% of vessel lumen diameter in all patients. Major amputation was performed in 16 patients; AP was <70 mm Hg in 4 patients and >or=70 mm Hg in 6. It was not practicable in the remaining 6 patients. Transcutaneous oxygen tension was <30 mm Hg in 15 patients and >or=30 mm Hg in 1 patient. For diagnosis of critical limb ischemia (CLI) in diabetic patients presenting with rest pain or foot ulcer, measurement of TcPO( 2) is essential not only when AP is not measurable but also when this value is >or=70 mm Hg.


A genetic roadmap to the response to genotoxic agents in human cells.
To maintain genome fidelity and prevent diseases such as cancer, our cells must constantly detect, and efficiently and precisely repair, DNA damage. Paradoxically, DNA-damaging agents in the form of radiation and chemotherapy are also used to treat cancer. Olivieri et al. used a CRISPR-based screen to identify genes that, when disrupted, lead to sensitivity or resistance to 27 different DNA-damaging agents used in the lab and/or in the clinic to treat cancer patients1. Their results reveal multiple new genes and connections that regulate these critical DNA damage repair pathways, with implications for basic and clinical research as well as cancer therapy.


Induction or protection from experimental autoimmune encephalomyelitis depends on the cytokine secretion profile of TCR peptide-specific regulatory CD4 T cells.
Autoimmune diseases can result from the breakdown of regulation and subsequent activation of self-antigenic determinant-reactive T cells. During the evolution of the autoimmune response to myelin basic protein (MBP) in B10.PL mice, several distinct T cell populations expand: the effectors mediating experimental autoimmune encephalomyelitis (EAE) are MBP-reactive, CD4+, and predominantly TCR Vbeta8.2+; in addition, at least two regulatory populations can be detected--one comprised of Vbeta14+ CD4 T cells, reactive to a framework region 3 determinant on the Vbeta8.2 chain, and a second that is CD8+ and reactive to another Vbeta8.2 determinant. The combined action of these two regulatory cell types controls disease-causing effectors, resulting in spontaneous recovery from disease. In this report, we reveal that the cytokine secretion pattern of TCR peptide-specific regulatory CD4 T cells can profoundly influence whether a type 1 or type 2 population predominates among MBP-specific CD4 effectors. The priming of type 1 regulatory T cells results in deviation of the Ag-specific effector T cell population in a type 2 direction and protection from disease. In contrast, induction of type 2 regulatory T cells results in exacerbation of EAE, poor recovery, and an increased frequency of type 1 effectors. Thus, the encephalitogenic potential of the MBP-reactive effector population is crucially and dominantly influenced by the cytokine secretion phenotype of regulatory CD4 T cells. These findings have important implications in understanding peripheral tolerance to self-Ags as well as in the design of TCR-based therapeutic approaches.


Using diffusion MRI data acquired with ultra-high gradient strength to improve tractography in routine-quality data.
The development of scanners with ultra-high gradient strength, spearheaded by the Human Connectome Project, has led to dramatic improvements in the spatial, angular, and diffusion resolution that is feasible for in vivo diffusion MRI acquisitions. The improved quality of the data can be exploited to achieve higher accuracy in the inference of both microstructural and macrostructural anatomy. However, such high-quality data can only be acquired on a handful of Connectom MRI scanners worldwide, while remaining prohibitive in clinical settings because of the constraints imposed by hardware and scanning time. In this study, we first update the classical protocols for tractography-based, manual annotation of major white-matter pathways, to adapt them to the much greater volume and variability of the streamlines that can be produced from today's state-of-the-art diffusion MRI data. We then use these protocols to annotate 42 major pathways manually in data from a Connectom scanner. Finally, we show that, when we use these manually annotated pathways as training data for global probabilistic tractography with anatomical neighborhood priors, we can perform highly accurate, automated reconstruction of the same pathways in much lower-quality, more widely available diffusion MRI data. The outcomes of this work include both a new, comprehensive atlas of WM pathways from Connectom data, and an updated version of our tractography toolbox, TRActs Constrained by UnderLying Anatomy (TRACULA), which is trained on data from this atlas. Both the atlas and TRACULA are distributed publicly as part of FreeSurfer. We present the first comprehensive comparison of TRACULA to the more conventional, multi-region-of-interest approach to automated tractography, and the first demonstration of training TRACULA on high-quality, Connectom data to benefit studies that use more modest acquisition protocols.


[Changes in the nuclei of the mammary glandular cells in the secretory cycle].
At the early stages of the secretory cycle, the activation of secretory cell nuclear chromatin is observed: the binding of acridine orange increases, while chromatin condensation in the peripheral zone of the nucleus decreases. The increase of the chromatin transcriptional activity provides for a significant increase in cytoplasmic RNA. Under the action of oxytocin reaction product granules are accumulated in conglomerates suggesting some changes in calcium distribution in the decondensed chromatin.


[Vaginal infections in a Family Health Clinic in the Metropolitan Region, Chile].
Vaginal infections are a frequent cause for consultation, but their prevalence and etiology vary in different populations. To determine the prevalence and etiologies of vaginal infection in women attending a family health center in the Metropolitan Region of Chile. The microbiological diagnosis was made by wet mount and Gram stain. Diagnosis of trichomoniasis was performed by wet mount, culture and polymerase chain reaction. 101 women aged 15-54, not selected by signs or symptoms of vaginal infection, 46 of them pregnant were included. In 47 women (46.5%), vaginal infections were diagnosed. An association was observed between age and frequency of vaginal infection. The proportion of infections among pregnant and non-pregnant women was similar. The most frequent infections were bacterial vaginosis (16.8%), vulvovaginal candidiasis (11.9%) and co-infections (6.9%). We found 5.9% of intermediate microbiota cases, 3% of trichomoniasis and 2% of aerobic vaginitis. Symptoms of vaginal infection had poor agreement with microbiological findings. Otherwise physical signs had good agreement with the presence of infection, but low to moderate concordance with a specific etiology. We found a high prevalence of vaginal infections in the study population. It is necessary to improve the definitions and criteria of microbiological diagnosis of co-infections and intermediate microbiota, for them to be diagnosed in the clinical practice. More descriptive questionnaires are recommended to enhance the usefulness of clinical examination.


Exercise Instructors for Older Adult Fitness: A Review of the Literature.
Group-exercise instructors are a vital social determinant of exercise enjoyment, attendance, and adherence. Instructors also affect the degree to which physical cultures are socially inclusive. In order to elucidate the roles that instructors play in affecting these outcomes, we conducted a scoping review. Scoping reviews are a preliminary method for assessing the breadth and depth of existing literature in order to identify key themes and gaps therein. Based on Arksey and O'Malley's (2005) framework, we identified 52 articles and book chapters, 33 of which were older-adult specific, using a university search engine that simultaneously searches multiple databases. We conceptually mapped the literature, which revealed instructors' vital roles as: (1) constructors of group social cohesion, (2) cultural intermediaries, (3) competent practitioners, (4) leaders and communicators, and (5) educators. Of these, the instructor's educative role lacks empirical attention. We conclude with implications for future research, practice, and policy.


Coronary subclavian steal syndrome.
Coronary subclavian steal syndrome (CSSS) is the reversal of blood flow in an internal mammary artery bypass graft that results in coronary ischemia. CSSS is an uncommon but treatable cause of coronary ischemia. In this review, we highlight the historical background and epidemiology of CSSS, common clinical presentations, diagnosis of CSSS and management strategies for relieving ischemia. We also present a case report to illustrate the complexity of CSSS and percutaneous management using current technology. Most commonly, CSSS results from atherosclerotic stenosis of the subclavian artery and occurs in 2.5-4.5% of patients referred for coronary artery bypass grafting (CABG). All patients referred for CABG should have bilateral noninvasive brachial blood pressures checked to screen for the underlying subclavian stenosis. A review of 98 case reports with 128 patients demonstrated a diverse clinical presentation of CSSS, including acute myocardial infarction, unstable angina and acute systolic heart failure. Resolution of CSSS symptoms has been reported with both surgical and percutaneous revascularization. Long-term patency with either revascularization strategy is excellent. Percutaneous revascularization is largely considered the first-line therapy for CSSS and can be safely performed prior to CABG to prevent CSSS. CSSS should be suspected in patients presenting with angina, heart failure or myocardial infarction after CABG. Successful amelioration of CSSS symptoms can be safely and effectively performed via percutaneous revascularization.


Back Extensor Strengthening Exercise and Backpack Wearing Treatment for Camptocormia in Parkinson's Disease: A Retrospective Pilot Study.
To investigate the effect of a conservative treatment regime in Parkinson's disease patients with camptocormia. Nine patients with Parkinson's disease were included in a retrospective pilot study of the value of back extensor strengthening exercise. Six inpatients received a 30-minute treatment, twice daily for 5 weeks, being treated on average for 34 days; while three outpatients visited the clinic and were educated for home exercise and backpack wearing treatment. Outpatients should be scheduled to visit the outpatient department to check physical status every 2-4 weeks for an average of 3 months. All patients except one showed statistically significant improvements in activities of daily living (ADL) and motor symptoms, as measured by flexion angle at standing posture, Unified Parkinson's Disease Rating Scale (UPDRS) II and III, and modified Hoehn-Yahr staging. Conservative treatment is effective in postural correction of camptocormia in Parkinson's disease, as well as improvement in ADL and motor symptoms.


[Structural-functional organization of Golgi apparatus].
This review is dedicated to the structure and function of Golgi apparatus (GA). It summarizes contemporary data published in numerous experimental papers and in several reviews. Possible ways of intra-Golgi transport of proteins, existent models of structural and functional organization of Golgi organelle, as well as the issues of its biogenesis, posttranslational modification and sorting of proteins and lipids, and mechanisms of their trafficking are discussed. Special attention is paid to the role of coatomer proteins (COPI, COPII and clathrin), fusion proteins (SNAREs), and small GTPases (ARF, SARI) in the secretory pathway. In addition, the phenomena of ultrastructural alterations of GA due to various functional conditions and physiological stimuli are specifically accented. We included in this review our original data on a probable involvement of GA in water transport, and on the organization of atypical GA in microsporidia--intracellular parasitic protists.


The direct influence of retinal degeneration on electrical stimulation efficacy: Significant implications for retinal prostheses.
Photoreceptor loss and inner retinal network remodeling severely impacts the ability of retinal prosthetic devices to create artificial vision. We developed a computational model of a degenerating retina based on rodent data and tested its response to retinal electrical stimulation. This model includes detailed network connectivity and diverse neural intrinsic properties, capable of exploring how the degenerated retina influences the performance of electrical stimulation during the degeneration process. Our model suggests the possibility of quantitatively modulating retinal ON and OFF pathways between phase II and III of retinal degeneration without requiring any differences between ON and OFF RGC intrinsic cellular properties. The model also provided insights about how remodeling events influence stage-dependent differential electrical responses of ON and OFF pathways.Clinical Relevance-This data-driven model can guide future development of retinal prostheses and stimulation strategies that may benefit patients at different stages of retinal disease progression, particularly in the early and mid-stages, thus increasing their global acceptance.


Organization of 5S genes in chromatin of Xenopus laevis.
The chromatin organization of the genes coding for 5S RNA in Xenopus laevis has been investigated with restriction endonucleases and micrococcal nuclease. Digestion of nuclei from liver, kidney, blood and kidney cells maintained in culture with micrococcal nuclease reveals that these Xenopus cells and tissues have shorter nucleosome repeat lengths than the corresponding cells and tissues from other higher organisms. 5S genes are organized in nucleosomes with repeat lengths similar to those of the bulk chromatin in liver (178 bp) and cultured cells (165 bp); however, 5S gene chromatin in blood cells has a shorter nucleosome repeat (176 bp) than the bulk of the genome in these cells (184 bp). From an analysis of the 5S DNA fragments produced by extensive restriction endonuclease cleavage of chromatin in situ, no special arrangement of the nucleosomes with respect to the sequence of 5S DNA can be detected. The relative abundance of 5S gene multimers follows a Kuhn distribution, with about 57% of all HindIII sites cleaved. This suggests that HindIII sites can be cleaved both in the nucleosome core and linker regions.


Comparison of infants and children with urolithiasis: a large case series.
We evaluated the demographic features, etiologic risk factors, treatment strategies, and outcome of the infants and children with urolithiasis (UL). A retrospective multicenter study was conducted including 23 Pediatric Nephrology centers in Turkey. The medical records of 2513 children with UL were reviewed. One thousand, three hundred and four boys and 1209 girls (1.1:1) were reported. The mean age at diagnosis was 39.5 ± 35 months (0.4-231 months), and 1262 patients (50.2%) were in the first year of life (infants). Most of the cases with infantile UL were diagnosed incidentally. Microlithiasis (< 3 mm) was found in 794 patients (31.6%), and 64.5% of the patients with microlithiasis were infants. Stones were located in the pelvis-calyces in 63.2% (n: 1530) of the cases. The most common stone type was calcium oxalate (64.6%). Hypocitraturia was the most common metabolic risk factor (MRF) in children older than 12 months, but in infancy, hypercalciuria was more common. Fifty-five percent of the patients had received at least one medical treatment, mostly potassium citrate. At the end of a year's follow-up, most of the patients with microlithiasis (85%) showed spontaneous remission. The rate of spontaneous stone resolution in infants was higher than in children. Spontaneous remission rate was higher in cases with MRF ( - ) stones than in MRF ( +) stones. However, remission rate with medical treatment was higher in cases with MRF ( +) stones. This study represents the results of a large series of infants and children with UL and showed that there are several differences such as underlying metabolic and anatomic abnormalities, clinical course, and stone remission rates between infants and children with urinary stone disease.


Long duration of atrial high-rate episode is more favorable in predicting ischemic stroke than high CHA2 DS2 -VASc score.
This study aimed to explore the roles of duration and burden of atrial high-rate episode (AHRE) on ischemic stroke in patients with pacemaker implantation. Patients with pacemaker implantation for bradycardia from 2013 to 2017 were consecutively enrolled. Data such as gender, age, combined diseases, type of AF, left atrial size, left ventricular size, left ventricular ejection fraction, CHA2 DS2 -VASc score, and anticoagulants were collected. The burden and duration of AHRE based on different interval partition were also recorded in detail to evaluate the impacts on ischemic stroke. Cox regression analysis with time-dependent covariates was conducted. A total of 220 patients with AHRE were enrolled. The average follow-up time was 48.42 ± 17.20 months. Univariate regression analysis showed that diabetes (p = .024), high CHA2 DS2 -VASc score (≥ 2) (p = .021), long mean AHRE burden (p = .011), long maximal AHRE burden (p = .015), long AHRE duration lasting≥48 h (p = .001) or 24 h (p = .001) or 12 h (p = .005) were prone to ischemic stroke. Further multivariate regression analysis showed that long duration of AHRE (≥48 h) (HR 10.77; 95% CI 3.22-55.12; p = .030) were significantly correlated with stroke in patients with paroxysmal AF. There was no significant correlation between the type of AF and stroke (p = .927). The longer duration of AHRE (≥48 h) was more favorable in predicting ischemic stroke than high CHA2 DS2 -VASc score (≥2).


Transfer of Bacillus thuringiensis plasmids coding for delta-endotoxin among strains of B. thuringiensis and B. cereus.
The recently discovered high-frequency transfer of plasmids between strains of Bacillus thuringiensis was used to study the genetic relationship between plasmids and production of the insecticidal delta-endotoxin crystal. Three strains of B. thuringiensis transmitted the Cry+ (crystal-producing) phenotype to Cry- (acrystalliferous) B. thuringiensis recipients. Agarose gel electrophoresis showed that one specific plasmid from each donor strain was always present in Cry+ "transcipients." The size of the transmissible crystal-coding plasmid varied with the donor strain, being 75 MDal (megadaltons) in size in HD-2, 50 MDal in HD-73, and 44 MDal in HD-263. Immunological analysis showed the Cry+ transcipients to be hybrid strains, having flagella of the recipient serotype and crystals of the donor serotype. These results demonstrate that the structural genes for the delta-endotoxin are plasmid borne. Crystal-coding plasmids also transferred into two strains of the related species Bacillus cereus and yielded transcipients that produced crystals of the same antigenicity as the donor strain.


The tunnel graft procedure for attachment of a hairpiece.
A new procedure for attaching a hairpiece to the scalp is described. Two skin-lined tunnels in the anterior and posterior scalp are built, using a strip of free full-thickness donor skin fron the postauricular area. Silicone-coated metal clips, suitably fashioned from .062'' Kirschner's wire, are then attached to the hairpiece. The metal clips are inserted into the skin-lined tunnels and thereby anchor the hairpiece to the scalp.


Doubly robust estimation of causal effects.
Doubly robust estimation combines a form of outcome regression with a model for the exposure (i.e., the propensity score) to estimate the causal effect of an exposure on an outcome. When used individually to estimate a causal effect, both outcome regression and propensity score methods are unbiased only if the statistical model is correctly specified. The doubly robust estimator combines these 2 approaches such that only 1 of the 2 models need be correctly specified to obtain an unbiased effect estimator. In this introduction to doubly robust estimators, the authors present a conceptual overview of doubly robust estimation, a simple worked example, results from a simulation study examining performance of estimated and bootstrapped standard errors, and a discussion of the potential advantages and limitations of this method. The supplementary material for this paper, which is posted on the Journal's Web site (http://aje.oupjournals.org/), includes a demonstration of the doubly robust property (Web Appendix 1) and a description of a SAS macro (SAS Institute, Inc., Cary, North Carolina) for doubly robust estimation, available for download at http://www.unc.edu/~mfunk/dr/.


Rac2 concentrations in umbilical cord neutrophils.
Human newborn infants display a variety of immunodeficiencies of immaturity, including diminished neutrophil adhesion, chemotaxis, and migration. Rac2, a guanosine triphosphate-binding protein, is an essential regulator of human neutrophil migration and chemotaxis. Since human subjects and mice deficient in Rac2 display deficiencies in neutrophil functions similar to newborn infants, we postulated that newborn neutrophils may be deficient in Rac2. The aim of the study was to measure Rac1 and Rac2 concentrations in neutrophils from umbilical cord blood. Neutrophils from cord and adult blood were isolated, total cell lysates extracted, and Rac protein concentrations determined using Western blot analysis. Rac2 concentrations were significantly lower in the neutrophil protein lysates isolated from cord blood compared to adult blood despite similar levels of Rac1. Diminished Rac2 expression in cord blood neutrophils may contribute to the defects observed in cord blood neutrophil function.


Recent hospitalization and the risk of hip fracture among older Americans.
We identified hip fracture risks in a prospective national study. Baseline (1993-1994) interview data were linked to Medicare claims for 1993-2005. Participants were 5,511 self-respondents aged 70 years and older and not in managed Medicare. ICD9-CM 820.xx (International Classification of Diseases, 9th Edition, Clinical Modification) codes identified hip fracture. Participants were censored at death or enrollment into managed Medicare. Static risk factors included sociodemographic, socioeconomic, place of residence, health behavior, disease history, and functional and cognitive status measures. A time-dependent marker reflecting postbaseline hospitalizations was included. A total of 495 (8.9%) participants suffered a postbaseline hip fracture. In the static proportional hazards model, the greatest risks involved age (adjusted hazard ratios [AHRs] of 2.01, 2.82, and 4.91 for 75-79, 80-84, and > or =85 year age groups vs those aged 70-74 years; p values <.001), sex (AHR = 0.45 for men vs women; p < .001), race (AHRs of 0.37 and 0.46 for African Americans and Hispanics vs whites; p values <.001 and <.01), body mass (AHRs of 0.40, 0.77, and 1.73 for obese, overweight, and underweight vs normal weight; p values <.001, <.05, and <.01), smoking status (AHRs = 1.49 and 1.52 for current and former smokers vs nonsmokers; p values <.05 and <.001), and diabetes (AHR = 1.99; p < .001). The time-dependent recent hospitalization marker did not alter the static model effect estimates, but it did substantially increase the risk of hip fracture (AHR = 2.51; p < .001). Enhanced discharge planning and home care for non-hip fracture hospitalizations could reduce subsequent hip fracture rates.


The Critical Role of Electrolyte Gating on the Hydrogen Evolution Performance of Monolayer MoS2.
According to density functional theory, monolayer (ML) MoS2 is predicted to possess electrocatalytic activity for the hydrogen evolution reaction (HER) that approaches that of platinum. However, its observed HER activity is much lower, which is widely believed to result from a large Schottky barrier between ML MoS2 and its electrical contact. In order to better understand the role of contact resistance in limiting the performance of ML MoS2 HER electrocatalysts, this study has employed well-defined test platforms that allow for the simultaneous measurement of contact resistance and electrocatalytic activity toward the HER during electrochemical testing. At open circuit potential, these measurements reveal that a 0.5 M H2SO4 electrolyte can act as a strong p-dopant that depletes free electrons in MoS2 and leads to extremely high contact resistance, even if the contact resistance of the as-made device in air is originally very low. However, under applied negative potentials this doping is mitigated by a strong electrolyte-mediated gating effect which can reduce the contact and sheet resistances of properly configured ML MoS2 electrocatalysts by more than 5 orders of magnitude. At potentials relevant to HER, the contact resistance becomes negligible and the performance of MoS2 electrodes is limited by HER kinetics. These findings have important implications for the design of low-dimensional semiconducting electrocatalysts and photocatalysts.


Gliosarcoma stem cells undergo glial and mesenchymal differentiation in vivo.
Cancer stem cells (CSCs) are characterized by their self-renewing potential and by their ability to differentiate and phenocopy the original tumor in orthotopic xenografts. Long-term propagation of glioblastoma (GBM) cells in serum-containing medium results in loss of the CSCs and outgrowth of cells genetically and biologically divergent from the parental tumors. In contrast, the use of a neurosphere assay, a serum-free culture for selection, and propagation of central nervous system-derived stem cells allows the selection of a subpopulation containing CSCs. Gliosarcoma (GS), a morphological variant comprising approximately 2% of GBMs, present a biphasic growth pattern, composed of glial and metaplastic mesenchymal components. To assess whether the neurosphere assay would allow the amplification of a subpopulation of cells with "gliosarcoma stem cell" properties, capable of propagating both components of this malignancy, we have generated neurospheres and serum cultures from primary GS and GBM surgical specimens. Neurosphere cultures from GBM and GS samples expressed neural stem cell markers Sox2, Musashi1, and Nestin. In contrast to the GBM neurosphere lines, the GS neurospheres were negative for the stem cell marker CD133. All neurosphere lines generated high-grade invasive orthotopic tumor xenografts, with histological features strikingly similar to the parental tumors, demonstrating that these cultures indeed are enriched in CSCs. Remarkably, low-passage GS serum cultures retained the expression of stem cell markers, the ability to form neurospheres, and tumorigenicity. The GS experimental tumors phenocopied the parental tumor, exhibiting biphasic glial and mesenchymal components, constituting a clinically relevant model to investigate mesenchymal differentiation in GBMs.


Hyperammonemic encephalopathy in an adenocarcinoma patient managed with carglumic acid.
Hyperammonemic encephalopathy (he) is a rare complication of malignancy and chemotherapy. Although the cause of he is unclear, a functional arginine deficiency secondary to increased catabolism has been suggested as a possible mechanism. Either that deficiency or an undetermined metabolite could lead to inhibition of N-acetylglutamate synthase (nags), a urea cycle enzyme, resulting in hyperammonemia. We present a case of chemotherapy-induced he in a patient with no underlying primary urea cycle disorder. The patient had a successful trial of carglumic acid (a synthetic analog of the product of nags), which suggests that, at least in some cases, he can be treated by overcoming proximal inhibition of the urea cycle. Further, our case is the first in the literature to exclude genetic defects and disorders of the proximal urea cycle, suggesting that hyperammonemia in these patients is probably secondary to chemotherapy.


The anergic state in sarcoidosis is associated with diminished dendritic cell function.
Sarcoidosis is a chronic inflammatory disease of unknown cause, characterized by granuloma formation similar to tuberculosis, but without clear evidence of a microbial infection. Because sarcoidosis is linked with clinical anergy and other evidence of diminished cellular immunity, we hypothesized that decreased skin delayed-type hypersensitivity (DTH) responses to recall Ags in affected individuals would be associated with decreased function of their blood dendritic cells (DCs). Our study involved ex vivo isolation, phenotyping, and functional testing of myeloid DCs (mDCs), plasmacytoid DCs, and T lymphocytes from blood of normal healthy volunteers and sarcoidosis subjects with active, untreated pulmonary disease. We found mDC function in the allogeneic MLR directly corresponded to the magnitude of skin DTH reactions to recall Ags in both sarcoidosis subjects and normal volunteers. However, both of these outcomes were significantly decreased in the sarcoidosis group. Diminished mDC function occurred despite up-regulated costimulatory and maturation markers. Clinical relevance is suggested by the inverse relationship between both mDC allogeneic responses and skin DTH responses with clinical disease severity as measured by chest radiograms. Because granulomas form when cellular immunity fails to clear antigenic stimuli, attenuated mDC function in sarcoidosis may contribute to susceptibility and persistence of the chronic inflammation characteristic of this disease.


Strong type 1, but impaired type 2, immune responses contribute to Orientia tsutsugamushi-induced pathology in mice.
Scrub typhus is a neglected, but important, tropical disease, which puts one-third of the world's population at risk. The disease is caused by Orientia tsutsugamushi, an obligately intracellular Gram-negative bacterium. Dysregulation in immune responses is known to contribute to disease pathogenesis; however, the nature and molecular basis of immune alterations are poorly defined. This study made use of a newly developed murine model of severe scrub typhus and focused on innate regulators and vascular growth factors in O. tsutsugamushi-infected liver, lungs and spleen. We found no activation or even reduction in base-line expression for multiple molecules (IL-7, IL-4, IL-13, GATA3, ROR-γt, and CXCL12) at 2, 6 and 10 days post-infection. This selective impairment in type 2-related immune responses correlated with a significant activation of the genes for IL-1β, IL-6, IL-10, TNF-α, IFN-γ, as well as CXCR3- and CXCR1-related chemokines in inflamed tissues. The elevated angiopoietin (Ang)-2 expression and Ang-2/Ang-1 ratios suggested excessive inflammation and the loss of endothelial integrity. These alterations, together with extensive recruitment of myeloperoxidase (MPO)-expressing neutrophils and the influx of CD3+ T cells, contributed to acute tissue damage and animal death. This is the first report of selective alterations in a panel of immune regulators during early O. tsutsugamushi infection in intravenously inoculated C57BL/6 mice. Our findings shed new light on the pathogenic mechanisms associated with severe scrub typhus and suggest potential targets for therapeutic investigation.


Neurofilament light chain and glial fibrillary acidic protein levels in metachromatic leukodystrophy.
Metachromatic leukodystrophy is a lethal metabolic leukodystrophy, with emerging treatments for early disease stages. Biomarkers to measure disease activity are required for clinical assessment and treatment follow-up. This retrospective study compared neurofilament light chain and glial fibrillary acidic protein (GFAP) levels in CSF (n = 11) and blood (n = 92) samples of 40 patients with metachromatic leukodystrophy (aged 0-42 years) with 38 neurologically healthy children (aged 0-17 years) and 38 healthy adults (aged 18-45 years), and analysed the associations between these levels with clinical phenotype and disease evolution in untreated and transplanted patients. Metachromatic leukodystrophy subtype was determined based on the (expected) age of symptom onset. Disease activity was assessed by measuring gross motor function deterioration and brain MRI. Longitudinal analyses with measurements up to 23 years after diagnosis were performed using linear mixed models. CSF and blood neurofilament light chain and GFAP levels in paediatric controls were negatively associated with age (all P < 0.001). Blood neurofilament light chain level at diagnosis (median, interquartile range; picograms per millilitre) was significantly increased in both presymptomatic (14.7, 10.6-56.7) and symptomatic patients (136, 40.8-445) compared to controls (5.6, 4.5-7.1), and highest among patients with late-infantile (456, 201-854) or early-juvenile metachromatic leukodystrophy (291.0, 104-445) and those ineligible for treatment based on best practice (291, 57.4-472). GFAP level (median, interquartile range; picogram per millilitre) was only increased in symptomatic patients (591, 224-1150) compared to controls (119, 78.2-338) and not significantly associated with treatment eligibility (P = 0.093). Higher blood neurofilament light chain and GFAP levels at diagnosis were associated with rapid disease progression in late-infantile (P = 0.006 and P = 0.051, respectively) and early-juvenile patients (P = 0.048 and P = 0.039, respectively). Finally, blood neurofilament light chain and GFAP levels decreased during follow-up in untreated and transplanted patients but remained elevated compared with controls. Only neurofilament light chain levels were associated with MRI deterioration (P < 0.001). This study indicates that both proteins may be considered as non-invasive biomarkers for clinical phenotype and disease stage at clinical assessment, and that neurofilament light chain might enable neurologists to make better informed treatment decisions. In addition, neurofilament light chain holds promise assessing treatment response. Importantly, both biomarkers require paediatric reference values, given that their levels first decrease before increasing with advancing age.


The cohort multiple randomized controlled trial design: a valid and efficient alternative to pragmatic trials?
Randomized controlled trials (RCTs)-the gold standard for evaluating the effects of medical interventions-are notoriously challenging in terms of logistics, planning and costs. The cohort multiple randomized controlled trial approach is designed to facilitate randomized trials for pragmatic evaluation of (new) interventions and is a promising variation from conventional pragmatic RCTs. In this paper, we evaluate methodological challenges of conducting an RCT within a cohort. We argue that equally valid results can be obtained from trials conducted within cohorts as from pragmatic RCTs. However, whether this design is more efficient compared with conducting a pragmatic RCT depends on the amount and nature of non-compliance in the intervention arm.


Southern Ocean sea-ice extent, productivity and iron flux over the past eight glacial cycles.
Sea ice and dust flux increased greatly in the Southern Ocean during the last glacial period. Palaeorecords provide contradictory evidence about marine productivity in this region, but beyond one glacial cycle, data were sparse. Here we present continuous chemical proxy data spanning the last eight glacial cycles (740,000 years) from the Dome C Antarctic ice core. These data constrain winter sea-ice extent in the Indian Ocean, Southern Ocean biogenic productivity and Patagonian climatic conditions. We found that maximum sea-ice extent is closely tied to Antarctic temperature on multi-millennial timescales, but less so on shorter timescales. Biological dimethylsulphide emissions south of the polar front seem to have changed little with climate, suggesting that sulphur compounds were not active in climate regulation. We observe large glacial-interglacial contrasts in iron deposition, which we infer reflects strongly changing Patagonian conditions. During glacial terminations, changes in Patagonia apparently preceded sea-ice reduction, indicating that multiple mechanisms may be responsible for different phases of CO2 increase during glacial terminations. We observe no changes in internal climatic feedbacks that could have caused the change in amplitude of Antarctic temperature variations observed 440,000 years ago.


Duration and regulation of the developmental cycle of Ixodes dammini (Acari: Ixodidae).
To determine how long the various developmental stages of the deer tick (Ixodes dammini Spielman, Clifford, Piesman & Corwin) survive in nature and to establish the interval between blood feeding and ecdysis or oviposition as well as subsequent larval eclosion, we observed ticks confined in the field. Unless adults feed during their first season (fall through spring) of activity, they die and do not survive the summer. Nonfed nymphs survive through two seasons (May through August) of feeding activity such that annual cohorts overlap. Nonfed larvae survive less than one year, and because they hatch toward the end of the summer, cohorts of this developmental stage do not overlap. Larvae that feed before September molt promptly and overwinter as nymphs; those that feed later overwinter engorged and ecdyse during the following spring. Fed nymphs fail to survive the winter, and thus must feed before late summer. They develop to the adult stage in the same year in which they feed. Regardless of time of feeding, females lay eggs in early summer, and the resulting larvae synchronously eclose late in that season. Although the cycle of this tick can be completed in nature in two years, it may extend to four years when hosts are relatively scarce. The seasonal inversion of larvae and nymphs appears to be regulated by physiological mechanisms and by host abundance.


Treatment and Resolution of Filamentary Keratitis in a Patient with Stiff Person Syndrome.
To report a rare case of filamentary keratitis associated with stiff person syndrome. Case report. A 26-year-old woman with several autoimmune conditions was referred because of filamentary keratitis. Conservative management using lubrication, mucolytics, and bandage contact lenses failed to adequately relieve symptoms. Despite the addition of oral prednisone, cyclosporine, and azathioprine, the patient's condition persisted with numerous filaments and severe dry eye. One month later, the patient had an episode of sudden muscle cramps in her back, for which she was hospitalized. Investigations revealed a diagnosis of stiff person syndrome, positive for glutamic acid decarboxylase autoantibodies. Plasmapheresis and high-dose intravenous steroids were provided as initial therapy. On follow-up, her filamentary keratitis resolved and at the last visit, her ocular symptoms had completely resolved. Filamentary keratitis is a chronic, recurrent condition that can be challenging to treat. Effective management requires careful consideration of possible predisposing causes, especially in refractory cases.


Types and frequency of errors in the preparation and administration of drugs.
To identify compatibility, types and frequency of errors in preparation and administration of intravenous drugs. A cross-sectional and descriptive study performed at the emergency department of a university hospital in the city of São Paulo (SP). The sample consisted of 303 observations of the preparation and administration of intravenous drugs by nursing aides, nursing technicians and registered nurses, using a systematized script, similar to a checklist. The following variables were collected: errors related to dispensing, omission, schedule, unauthorized administration, dosage, formulation, incompatibility, preparation and administration. In the preparation stage, the following errors were identified: no hand hygiene (70.29%), and no use of aseptic technique (80.85%). Upon administration, no hand hygiene (81.18%), and no use of aseptic technique (84.81%). In 31.35% of observations, there was more than one medication at the same time for the same patient, of which 17.89% were compatible, 56.84% were incompatible and 25.26% were not tested, according to the Micromedex database. In both preparation and administration stages, the most frequent errors were no hand hygiene and no use of aseptic technique, indicating the need to develop and implement education programs focused on patient safety.


Thermal limits on MV x-ray production by bremsstrahlung targets in the context of novel linear accelerators.
To study the impact of target geometrical and linac operational parameters, such as target material and thickness, electron beam size, repetition rate, and mean current on the ability of the radiotherapy treatment head to deliver high-dose-rate x-ray irradiation in the context of novel linear accelerators capable of higher repetition rates/duty cycle than conventional clinical linacs. The depth dose in a water phantom without a flattening filter and heat deposition in an x-ray target by 10 MeV pulsed electron beams were calculated using the Monte-Carlo code MCNPX, and the transient temperature behavior of the target was simulated by ANSYS. Several parameters that affect both the dose distribution and temperature behavior were investigated. The target was tungsten with a thickness ranging from 0 to 3 mm and a copper heat remover layer. An electron beam with full width at half maximum (FWHM) between 0 and3 mm and mean current of 0.05-2 mA was used as the primary beam at repetition rates of 100, 200, 400, and 800 Hz. For a 10 MeV electron beam with FWHM of 1 mm, pulse length of 5 μs, by using a thin tungsten target with thickness of 0.2 mm instead of 1 mm, and by employing a high repetition rate of 800 Hz instead of 100 Hz, the maximum dose rate delivered can increase two times from 0.57 to 1.16 Gy/s. In this simple model, the limiting factor on dose rate is the copper heat remover's softening temperature, which was considered to be 500°C in our study. A high dose rate can be obtained by employing thin targets together with high repetition rate electron beams enabled by novel linac designs, whereas the benefit of thin targets is marginal at conventional repetition rates. Next generation linacs used to increase dose rate need different target designs compared to conventional linacs.


[Characterization of an endogenous factor with digitalic activity in the guinea pig heart (II): Binding to a receptor].
In this work we continue the studies on the presence of an endogenous digitalis-like factor in mammals. A water extract from guinea pig heart was partially purified by liquid chromatography. The extract displaced (3H)ouabain specific binding to guinea pig heart Na,K-ATPase yielding a displacement curve parallel to that observed when digoxin was used as displacing agent. The data suggest that the activity of the extract was due to the presence of a factor that recognizes the digitalis binding site but not to ions nor lipids. Furthermore, the extract cross-reacted with antidigoxin antibodies in a manner similar to the specific antigens. These results support the hypothesis of the existence of an endogenous ligand of the digitalis receptor in mammals that could be a physiological regulator of the sodium pump.


[The intermediate nerve and its place in the system of cranial nerves].
The intermediate nerve (IN) in embryogenesis of man, cat and white rat is formed similarly. From the common with the VIII cranial nerve anlage the geniculum node, the vestibular and cochlear ganglia are emerged. A separated root of the IN connects the geniculum node with the nuclei, situating in the CNS. From the geniculum node main branches of the IN get off: the greater petrosal nerve and the cord of the tympanum, its fibers in the temporal bone canal run in the facial nerve trunk. In the periphery the IN branches unite with other cranial nerves, make connections with branches of vegetative (parasympathetic) ganglia and plexuses. They spread along a large territory, forming a system of parasympathetic innervation in the area of the head and ensuring with specific nervous apparatuses the gustatory organ. It is expedient to select the IN into an independent cranial nerve and confirm it the number of the regular pair.


Steady-state kinetics of mouse DNA polymerase alpha.
Steady-state kinetic studies of DNA polymerase alpha purified from mouse myeloma MOPC104E cells have been carried out. The results of initial velocity analysis with or without sodium pyrophosphate, a product inhibitor, indicated that the reaction mechanism of this enzyme can be categorized as an ordered Bi Bi type where the concentration of the ternary complex is very low.


The road to herbaria: Teaching and learning about biology, aesthetics, and the history of botany.
Having recently published a book on herbaria, collections of preserved plants, I look back on my career teaching biology and the experiences that led me to the world of dried plants. My first job was teaching non-science majors, to which I devoted 45 years. Over that time, I worked to make biological inquiry as fascinating to my students as it was to me. I found that the private side of science, how science is actually done, was a good entry point with stories about the wrong turns, difficulties, and joys of research. This led me to biographies and essays by biologists, works on the philosophy and history of science, and interest in the relationship between science and art. Eventually these avenues brought me to the aesthetics of biology and then to botanical art and finally to where these streams seemed to come together in herbaria. I literally fell in love with them. Specimens contain a rich history of people and places, of exploration, colonization, exploitation, and the development of modern botany. Herbaria today are key to documenting and conserving the earth's biodiversity and tracing how climate change is shaping it. Through digitization, herbaria are an essential component of bioinformatics.


Gastrointestinal toxicity during induction treatment for childhood acute lymphoblastic leukemia: The impact of the gut microbiota.
Intestinal mucositis is a common side effect of chemotherapy leading to diarrhea, abdominal pain and increased risk of infections. The intestinal microbiota has been recognized as a key regulator of mucosal immune responses. Therefore, we hypothesized that intestinal microbial changes would be associated with enterocyte loss and systemic inflammation during induction treatment for childhood acute lymphoblastic leukemia (ALL). We prospectively included 51 children newly-diagnosed with ALL treated in Denmark in 2015-2018. Plasma C-reactive protein (CRP), plasma citrulline (marker of functional enterocytes mass) measurements and fecal samplings were performed on treatment Days 1, 8, 15, 22 and 29. Moreover, intestinal mucositis was scored by a trained nurse/physician. Fecal samples in patients and 19 healthy siblings were analyzed by 16S rRNA gene sequencing (V3-V4 region). Bacterial alpha diversity was lower in patients compared to siblings. It decreased from Day 1 to Days 8-22 and increased on Day 29. Shannon alpha diversity index was correlated with CRP on Days 15-29 (rho = -0.33-0.49; p < 0.05) and with citrulline on Days 15 and 29 (although with p values <0.06, rho = 0.32-0.34). The abundance of unclassified Enterococcus species (spp.) was correlated with CRP on Days 22-29 (rho = 0.42-0.49; p < 0.009), while the abundance of unclassified Lachnospiraceae spp. was correlated with citrulline on days 8-15 (rho = 0.48-0.62, p < 0.001). Systemic inflammation, enterocyte loss and relative abundance of unclassified Enterococcus spp. reached a peak around Day 15. In conclusion, specific changes in the microbiota were associated with the severity of enterocyte loss and systemic inflammation during chemotherapy.


Association between genetics of diabetes, coronary artery disease, and macrovascular complications: exploring a common ground hypothesis.
Type 2 diabetes and coronary artery disease (CAD) are conditions that cause a substantial public health burden. Since both conditions often coexist in the same individual, it has been hypothesized that they have a common effector. Insulin and hyperglycemia are assumed to play critical roles in this scenario. In recent years, many genetic risk factors for both diabetes and CAD have been discovered, mainly through genome-wide association studies. Genetic aspects of diabetes, diabetic macrovascular complications, and CAD are assumed to have intersections leading to the common effector hypothesis. However, only a few genetic risk factors could be identified that modulate the risk for both conditions. Polymorphisms in TCF7L2 and near the CDKN2A/B genes seem to be of great importance in this regard since they appear to modulate both conditions, and they are not necessarily related to insulinism, or hyperglycemia, for CAD development. Other issues related to this hypothesis, such as the problems of phenotype heterogeneity, are also of interest. Recent studies have contributed to a better understanding of the complex genetics of diabetic macrovascular complications. Much effort is still needed to clarify the associations in the genetics of diabetes, and cardiovascular disease. At present, there is little genetic evidence to support a common effector hypothesis, other than insulin or hyperglycemia, for the association between these conditions.


Liver steatosis (LS) evaluated through chemical-shift magnetic resonance imaging liver enzymes in morbid obesity; effect of weight loss obtained with intragastric balloon gastric banding.
The aim of this study was to evaluate in morbid obesity clinical and metabolic effects related to weight loss on liver steatosis (LS), measured through chemical-shift magnetic resonance imaging (MRI) and liver enzymes. Forty obese subjects (8 M/32 W; BMI 42.8 ± 7.12 kg/m(2), mean ± SD) were evaluated for LS through ultrasound (US-LS), chemical-shift MRI (MRI-LS), liver enzymes [aspartate aminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyltransferase (GGT), alkaline phosphatase (ALP)], anthropometric parameters [weight, BMI, waist circumference (WC)], lipids, insulin, insulin resistance (HOMA-IR), glycated hemoglobin (HbA1c), oral glucose tolerance test, and body composition [fat mass (FM) and fat-free mass (FFM) at bio-impedance analysis (BIA)]. Anthropometric measures, MRI-LS, BIA, and biochemical parameters were reevaluated 6 months later in 18 subjects undergoing restrictive bariatric approach, i.e., intragastric balloon (BIB, n = 13) or gastric banding (LAGB, n = 5), and in 13 subjects receiving hypocaloric diet. At baseline, US-LS correlates only with MRI-LS, and the latter correlates with ALT, AST, and GGT. After 6 months, subjects undergoing BIB or LAGB had significant changes of BMI, weight, WC, ALT, AST, GGT, ALP, HbA1c, insulin, HOMA-IR, FM, FFM, and MRI-LS. Diet-treated obese subjects had no significant change of any parameter under study; change of BMI, fat mass, and fat-free mass was significantly greater in LAGB/BIB subjects than in diet-treated subjects. Change of MRI-LS showed a significant correlation with changes in weight, BMI, WC, GGT, ALP, and basal MRI-LS. Significant weight loss after BIB or LAGB is associated with decrease in chemical-shift MRI-LS and with reduction in liver enzymes; chemical-shift MRI and liver enzymes allow monitoring of LS in follow-up studies.


HIV in adolescents in sub-Saharan Africa.
In this review, we summarize existing evidence on the effectiveness of different intervention approaches to HIV prevention in adolescents - focusing on studies that are either from or are relevant to sub-Saharan Africa. In addition, we include a brief review of other salient issues relevant to HIV prevention research in adolescents. Although numerous adolescent behavioural HIV prevention interventions have been evaluated, few have assessed their impact on HIV endpoints or been undertaken in Africa. In the three trials from Africa, which had HIV endpoints, none of the interventions had an impact on HIV, although all affected some knowledge and attitudes and reported behaviours. In one of these trials, there was a borderline effect on herpes simplex virus-2 incidence. Adolescents have typically been excluded from trials of biological interventions, although they are likely to benefit from these interventions if found to be effective. Despite the regulatory difficulties, they must be considered for inclusion in these trials as an important target population. Although structural determinants of infection appear to be key drivers of the epidemic in young people in sub-Saharan Africa, few have been rigorously evaluated; those that have have been shown to be promising evidence for their future role in prevention. Young people in sub-Saharan Africa, particularly young women, continue to bear the brunt of the HIV epidemic. Adolescents must be an important focus for HIV prevention programming and research. It is increasingly clear that multilevel approaches to prevention will be required to reduce rates of HIV in this age group.


Drug-induced endocytosis and entrapment in red cells and ghosts.
Induction of endocytosis in intact human RBC can be achieved by treating RBC in vitro with several classes of amphipathic cations. Perhaps 1-2% of the 90 fl volume of the red cell will then consist of any material suspended or dissolved in the bulk supernatant medium. This amount could be increased if the material in question also binds selectively to the outer surface of the red cell's plasma membrane. Such a red cell, now rendered spherostomatocytic if reinjected into the circulation, would be removed by the monocytes-macrophages concentrated in spleen and to a lesser extent in liver, lymph nodes, and bone marrow. Conceivably, one could deliver a significant amount of drug or other agent to monocytes-macrophages in these sites. Endocytosis can also be induced in white ghosts and the resulting vacuoles are not accessible to small molecules. The targeting of white ghosts has not been subjected to extensive study but such ghosts containing material included in their endocytic vacuoles could be used to deliver agents to the recipient's monocyte-macrophage system.


In vitro studies, pharmacokinetic studies and clinical use of a high purity double virus inactivated FVIII/VWF concentrate (Immunate) in the treatment of von Willebrand disease.
Patients with type 2 and 3 von Willebrand disease (VWD) are treated with factor VIII/VWF concentrate in case of bleeding or surgery. Immunate (Baxter, Vienna, Austria) is a double virus inactivated FVIII/VWF concentrate and is registered in several countries for patients with VWD with reduced FVIII levels. We performed an in vitro, a pharmacokinetic and a clinical study to evaluate Immunate in VWD. In vitro studies showed a significant variation in VWF levels in 9 different batches. The median (range) values (in IU/mL) were 1.10 (0.98-1.30) for FVIII:C, 1.34 (0.95-1.61) for VWF:Ag, 0.60 (0.27-1.08) for VWF:CBA and 0.73 (0.59-0.94) for VWF:RCo. The relatively low VWF activity is mainly due to the lack of high molecular weight multimers (HMWM), as determined by electrophoresis. A pharmacokinetic study showed, based on a content of FVIII:C of 1 U/mL, in vivo recoveries (%) of 106 (56-150) (median and range) for FVIII:C, 105 (62-187) for VWF:Ag, 25 (7-41) for VWF:CBA and 43 (11-76) for VWF:RCo. Half-lives were 14.1 h (7.4-36.9) for FVIII:C, 10.8 h (7.7-26.2) for VWF:Ag, 15.3 h (7.8-44.6) for VWF:CBA and 16.4 h (4.2-26.5) for VWF:RCo. In a clinical study efficacy was determined after infusion given before surgery or dental extractions in ten patients. In two patients the hemostatic response was classified as inadequate. In conclusion, there is a wide variability in VWF concentration and activity in various batches of Immunate. In the clinical study in which the dosage was based on FVIII:C contents of the concentrate, two out of ten patients had an insufficient haemostatic response. Therefore dosing of Immunate dosing should not be based on FVIII:C levels, but should be based on VWF activity of the individual batches. Future studies using a VWF activity-guided dosage regimen have to be performed to establish the efficacy of Immunate in the treatment of von Willebrand disease.


Characterization of pFOX-7a, a conjugative IncL/M plasmid encoding the FOX-7 AmpC-type β-lactamase, involved in a large outbreak in a neonatal intensive care unit.
FOX-type enzymes are a lineage of AmpC-type β-lactamases from Aeromonas spp. whose genes have been mobilized to plasmids spreading among Enterobacteriaceae, where they can be responsible for resistance to extended-spectrum cephalosporins and β-lactamase inhibitor combinations. Little is known about the genetic context and plasmid vehicles of bla(FOX) determinants. Here, we have characterized a plasmid encoding the FOX-7 β-lactamase, which was involved in a large outbreak caused by two Klebsiella pneumoniae clones in a neonatal intensive care unit. Plasmid transferability was tested in conjugation experiments using Escherichia coli recipients. Plasmids from different strains were compared by restriction profiling and PCR mapping. The complete sequence of pFOX-7a plasmid was determined by a next-generation sequencing approach followed by gap filling using PCR and sequencing. An apparently identical conjugative plasmid encoding FOX-7 was detected in representatives of the K. pneumoniae clones that caused the outbreak and in sporadic FOX-7-producing strains of other species from the same ward. The plasmid, named pFOX-7a, has an IncL/M-type backbone and two separate resistance modules including a Tn3-like transposon and a novel Tn1696 derivative, named Tn6234, which carries an integron platform, a hybrid (but still functional) mercury resistance module and a novel putative transposon of original structure, named Tn6240, associated with the bla(FOX-7) gene. pFOX-7a is the first completely characterized plasmid encoding a FOX-type β-lactamase. The bla(FOX-7) gene was associated with a putative transposable element of original structure, which was likely involved in its mobilization from the Aeromonas metagenome.


Evaluation of the effectiveness of air pollution control measures in Hong Kong.
From 2005 to 2013, volatile organic compounds (VOCs) and other trace gases were continuously measured at a suburban site in Hong Kong. The measurement data showed that the concentrations of most air pollutants decreased during these years. However, ozone (O3) and total non-methane hydrocarbon levels increased with the rate of 0.23 ± 0.03 and 0.34 ± 0.02 ppbv/year, respectively, pointing to the increasing severity of photochemical pollution in Hong Kong. The Hong Kong government has ongoing programs to improve air quality in Hong Kong, including a solvent program implemented during 2007-2011, and a diesel commercial vehicle (DCV) program since 2007. From before to after the solvent program, the sum of toluene, ethylbenzene and xylene isomers decreased continuously with an average rate of -99.1 ± 6.9 pptv/year, whereas the sum of ethene and propene increased by 48.2 ± 2.0 pptv/year from before to during the DCV program. Despite this, source apportionment results showed that VOCs emitted from diesel exhaust decreased at a rate of -304.5 ± 17.7 pptv/year, while solvent related VOCs decreased at a rate of -204.7 ± 39.7 pptv/year. The gasoline and liquefied petroleum gas vehicle emissions elevated by 1086 ± 34 pptv/year, and were responsible for the increases of ethene and propene. Overall, the simulated O3 rate of increase was lowered from 0.39 ± 0.03 to 0.16 ± 0.05 ppbv/year by the solvent and DCV programs, because O3 produced by solvent usage and diesel exhaust related VOCs decreased (p < 0.05) by 0.16 ± 0.01 and 0.05 ± 0.01 ppbv/year between 2005 and 2013, respectively. However, enhanced VOC emissions from gasoline and LPG vehicles accounted for most of the O3 increment (0.09 ± 0.01 out of 0.16 ± 0.05 ppbv/year) in these years. To maintain a zero O3 increment in 2020 relative to 2010, the lowest reduction ratio of VOCs/NOx was ∼1.5 under the NOx reduction of 20-30% which was based on the emission reduction plan for Pearl River Delta region in 2020.


Rembrandt: helping personalized medicine become a reality through integrative translational research.
Finding better therapies for the treatment of brain tumors is hampered by the lack of consistently obtained molecular data in a large sample set and the ability to integrate biomedical data from disparate sources enabling translation of therapies from bench to bedside. Hence, a critical factor in the advancement of biomedical research and clinical translation is the ease with which data can be integrated, redistributed, and analyzed both within and across functional domains. Novel biomedical informatics infrastructure and tools are essential for developing individualized patient treatment based on the specific genomic signatures in each patient's tumor. Here, we present Repository of Molecular Brain Neoplasia Data (Rembrandt), a cancer clinical genomics database and a Web-based data mining and analysis platform aimed at facilitating discovery by connecting the dots between clinical information and genomic characterization data. To date, Rembrandt contains data generated through the Glioma Molecular Diagnostic Initiative from 874 glioma specimens comprising approximately 566 gene expression arrays, 834 copy number arrays, and 13,472 clinical phenotype data points. Data can be queried and visualized for a selected gene across all data platforms or for multiple genes in a selected platform. Additionally, gene sets can be limited to clinically important annotations including secreted, kinase, membrane, and known gene-anomaly pairs to facilitate the discovery of novel biomarkers and therapeutic targets. We believe that Rembrandt represents a prototype of how high-throughput genomic and clinical data can be integrated in a way that will allow expeditious and efficient translation of laboratory discoveries to the clinic.


Bioaccumulation of heavy metals in Spartina.
The Spartina Schreb. genus is composed of C4 perennial grasses in the family Poaceae. They are native to the coasts of the Atlantic Ocean in western and southern Europe, north-west and southern Africa, the Americas and the southern Atlantic Ocean islands. Most species are salt tolerant and colonise coastal or inland saltmarshes. The available literature on heavy metal bioaccumulation by Spartina sp. was compiled and compared. Spartina alterniflora Loisel. and Spartina maritima (Curtis) Fernald were the most commonly researched species of the genus, whereas many species were not represented at all. In contrast, Cu and Zn are the most intensively researched heavy metals. The few studies dealing with the physiological impacts of heavy metals or the mechanisms of metal accumulation, which involve extracellular and intracellular metal chelation, precipitation, compartmentalisation and translocation in the vascular system, were documented. Bioaccumulation of metals in roots and tillers of some species of the Spartina genus (e.g. S. maritima and Spartina densiflora Brongn.) has been described as a feasible method for remediating waters and soils contaminated with heavy metals. One such example is Spartina argentinensis Parodi, which has been found to be a Cr-hyperaccumulator; it can concentrate chromium in its tissues to levels far exceeding those present in the soil.


Reactions templated by nucleic acids: more ways to translate oligonucleotide-based instructions into emerging function.
The programmability of oligonucleotide recognition offers an attractive platform to direct the assembly of reactive partners that can engage in chemical reactions. Recently, significant progress has been made in both the breadth of chemical transformations and in the functional output of the reaction. Herein we summarize these recent progresses and illustrate their applications to translate oligonucleotide instructions into functional materials and novel architectures (conductive polymers, nanopatterns, novel oligonucleotide junctions); into fluorescent or bioactive molecule using cellular RNA; to interrogate secondary structures or oligonucelic acids; or a synthetic oligomer.


Measurement of serum L-asparagine in the presence of L-asparaginase requires the presence of an L-asparaginase inhibitor.
The antileukemic activity of L-asparaginase (ASNase), an important component of therapy for acute lymphoblastic leukemia, is thought to result from depletion of serum L-asparagine (Asn). In studies of the pharmacological effects of ASNase, investigators have reported prolonged reduction in the serum concentration of Asn after the administration of ASNase. Such measurements may not be valid because ASNase present in the blood sample may hydrolyze Asn before its determination. We examined recovery of [U-14C]Asn from blood samples with and without various concentrations of added ASNase. In the presence of greater than or equal to 0.01 IU/ml of ASNase, the amount of [U-14C]Asn recovered was less than 15% of that without ASNase. Utilizing this assay, we studied the effect of 2 known inhibitors of ASNase in an attempt to improve Asn recovery. In the presence of aspartic beta semialdehyde (ASA), or 5-diazo-4-oxo-L-norvaline (DONV), and up to 1.0 IU/ml ASNase, Asn levels remained at greater than 90% of control. ASA prevented the hydrolysis of exogenous Asn in blood samples drawn from patients after ASNase injection. We also developed a method to determine Asn in serum utilizing high pressure liquid chromatography. Using this method, we found that the Asn level was greater than 90% of a normal level in the presence of 40 mM DONV and 1.0 IU/ml ASNase. Examination of serum from 4 patients treated with ASNase showed that Asn is detectable 7-19 days sooner when DONV is present in the blood collection system than in its absence. We conclude that: (a) as little as 0.01 IU/ml ASNase can hydrolyze Asn added to blood; (b) continued hydrolysis of Asn by ASNase ex vivo can result in falsely low serum Asn measurements; (c) ASA or DONV present in the collection tubes obviates the problem of continued ASNase activity; and (d) the degree and duration of Asn depletion after ASNase therapy is much less than previously believed. Thus, for accurate measurements of the duration and degree of Asn depletion by ASNase, an ASNase inhibitor such as ASA or DONV should be present in the blood collection system.


Aggressive strategic planning for oral health in Kuwait: a decade of post-war successes.
Strategic planning and implementation of oral health care and disease prevention programmes after the 1990/91 Gulf war are discussed. The key concept was to develop access to care and disease prevention for all Kuwaiti children in government kindergarten/primary schools and to eliminate emphasis on extractions and restorations. Resources were restored to pre-war levels and then increased. Prevention programmes for 150 000 children were established. Prevention funds increased from 7% to 20% of the oral health budget. Prevention-based dentists increased from 9.7% to 28.0% of staff. Rising caries trends were stabilized or reduced by up to 36.8%. Percentage of caries-free primary dentition in children increased up to 37.6%, permanent dentition up to 27.0%. A dentistry school was established.


On-chip refractive index cytometry for whole-cell deformability discrimination.
On-chip high-throughput phenotyping of single cells has gained a lot of interest recently due to the discrimination capability of label-free biomarkers such as whole-cell deformability and refractive index. Here we present on-chip refractive index cytometry (RIC) for whole-cell deformability at a high measurement rate. We have further exploited a previously published on-chip optical characterization method which enhances cellular discrimination through the refractive index measurement of single cells. The proposed on-chip RIC can simultaneously probe the cellular refractive index, effective volume and whole-cell deformability while reaching a measurement rate up to 5000 cells per second. Additionally, the relative position of the nucleus inside the cell is reflected by the asymmetry of the measured curve. This particular finding is confirmed by our numerical simulation model and emphasized by a modified cytoskeleton HL-60 cells model. Furthermore, the proposed device discriminated HL-60 derived myeloid cells such as neutrophils, basophils and promyelocytes, which are indistinguishable using flow cytometry. To our knowledge, this is the first integrated device to simultaneously characterize the cellular refractive index and whole-cell deformability, yielding enhanced discrimination of large myeloid cell populations.


A and H blood group antigens as markers of sucrase-isomaltase from the enterocyte-like differentiated human colon carcinoma cell lines HT-29 and Caco-2.
The purpose of this work was to investigate whether sucrase-isomaltase from enterocyte-like differentiated human colon carcinoma cell lines carries blood group antigens of the ABH system. Six cultured lines of blood group A (HT-29, SW-480, Co-115) or O phenotype (Caco-2, HRT-18, HCT-8R) were studied. Only HT-29 cells grown in the absence of glucose (HT-29 Glc-) and Caco-2 cells express an enterocytic differentiation with the presence of sucrase-isomaltase on the apical surface of the cells. Binding of anti-A antibodies to HT-29 Glc- and of UEA-I to Caco-2 cells gave the same apical immunofluorescence pattern of staining as did anti-sucrase-isomaltase antibodies, whereas only a membrane binding was observed in nondifferentiated cells. Sucrase-isomaltase immunoisolated from HT-29 Glc- and Caco-2 cells reacted with anti-A antibodies and Ulex europaeus agglutinin-I (UEA-I), respectively, at sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot. Immunoprecipitation of solubilized brush border-enriched fractions from the same cells with UEA-I or anti-A antibodies resulted in an inhibition of sucrase activity which reached congruent to 80% for Caco-2 cells with UEA-I and approximately equal to 50% for HT-29 cells with anti-A antibodies. Similar results were obtained in the corresponding tumors in nude mice: anti-A antibodies in HT-29 and UEA-I in Caco-2 tumors bound to the same apical structures as did anti-sucrase-isomaltase antibodies; sucrase-isomaltase immunoisolated from the tumors bound anti-A antibodies (HT-29) or UEA-I (Caco-2). These results support the hypothesis that sucrase-isomaltase from enterocyte-like differentiated human colon cancer cells carries blood group antigens of the ABH system. These findings suggest that colon cancers which have been shown to display an apical pattern of expression of ABH antigens should be screened for their possible enterocytic differentiation.