Long-term Recurrence and Complications Associated With Elective Incisional Hernia Repair.
Prosthetic mesh is frequently used to reinforce the repair of abdominal wall incisional hernias. The benefits of mesh for reducing the risk of hernia recurrence or the long-term risks of mesh-related complications are not known. To investigate the risks of long-term recurrence and mesh-related complications following elective abdominal wall hernia repair in a population with complete follow-up. Registry-based nationwide cohort study including all elective incisional hernia repairs in Denmark from January 1, 2007, to December 31, 2010. A total of 3242 patients with incisional repair were included. Follow-up until November 1, 2014, was obtained by merging data with prospective registrations from the Danish National Patient Registry supplemented with a retrospective manual review of patient records. A 100% follow-up rate was obtained. Hernia repair using mesh performed by either open or laparoscopic techniques vs open repair without use of mesh. Five-year risk of reoperation for recurrence and 5-year risk of all mesh-related complications requiring subsequent surgery. Among the 3242 patients (mean age, 58.5 [SD, 13.5] years; 1720 women [53.1%]), 1119 underwent open mesh repair (34.5%), 366 had open nonmesh repair (11.3%), and 1757 had laparoscopic mesh repair (54.2%). The median follow-up after open mesh repair was 59 (interquartile range [IQR], 44-80) months, after nonmesh open repair was 62 (IQR, 44-79) months, and after laparoscopic mesh repair was 61 (IQR, 48-78) months. The risk of the need for repair for recurrent hernia following these initial hernia operations was lower for patients with open mesh repair (12.3% [95% CI, 10.4%-14.3%]; risk difference, -4.8% [95% CI, -9.1% to -0.5%]) and for patients with laparoscopic mesh repair (10.6% [95% CI, 9.2%-12.1%]; risk difference, -6.5% [95% CI, -10.6% to -2.4%]) compared with nonmesh repair (17.1% [95% CI, 13.2%-20.9%]). For the entirety of the follow-up duration, there was a progressively increasing number of mesh-related complications for both open and laparoscopic procedures. At 5 years of follow-up, the cumulative incidence of mesh-related complications was 5.6% (95% CI, 4.2%-6.9%) for patients who underwent open mesh hernia repair and 3.7% (95% CI, 2.8%-4.6%) for patients who underwent laparoscopic mesh repair. The long-term repair-related complication rate for patients with an initial nonmesh repair was 0.8% (open nonmesh repair vs open mesh repair: risk difference, 5.3% [95% CI, 4.4%-6.2%]; open nonmesh repair vs laparoscopic mesh repair: risk difference, 3.4% [95% CI, 2.7%-4.1%]). Among patients undergoing incisional repair, sutured repair was associated with a higher risk of reoperation for recurrence over 5 years compared with open mesh and laparoscopic mesh repair. With long-term follow-up, the benefits attributable to mesh are offset in part by mesh-related complications.


Dynamic generation of robust and controlled beating signals in an asymmetric procedure of light storage and retrieval.
We propose an efficient scheme for the robust and controlled generation of beating signals in a sample of stationary atoms driven into the tripod configuration. This scheme relies on an asymmetric procedure of light storage and retrieval where the two classical coupling fields have equal detunings in the storage stage but opposite detunings in the retrieval stage. A quantum probe field, incident upon such an atomic sample, is first transformed into two spin coherence wave-packets and then retrieved with two optical components characterized by different time-dependent phases. Therefore the retrieved quantum probe field exhibits a series of maxima and minima (beating signals) in intensity due to the alternative constructive and destructive interference. This interesting phenomenon involves in fact the coherent manipulation of two dark-state polaritons and may be explored to achieve the fast quantum limited measurement.


Enhancing the detection of barcoded reads in high throughput DNA sequencing data by controlling the false discovery rate.
DNA barcodes are short unique sequences used to label DNA or RNA-derived samples in multiplexed deep sequencing experiments. During the demultiplexing step, barcodes must be detected and their position identified. In some cases (e.g., with PacBio SMRT), the position of the barcode and DNA context is not well defined. Many reads start inside the genomic insert so that adjacent primers might be missed. The matter is further complicated by coincidental similarities between barcode sequences and reference DNA. Therefore, a robust strategy is required in order to detect barcoded reads and avoid a large number of false positives or negatives.For mass inference problems such as this one, false discovery rate (FDR) methods are powerful and balanced solutions. Since existing FDR methods cannot be applied to this particular problem, we present an adapted FDR method that is suitable for the detection of barcoded reads as well as suggest possible improvements. In our analysis, barcode sequences showed high rates of coincidental similarities with the Mus musculus reference DNA. This problem became more acute when the length of the barcode sequence decreased and the number of barcodes in the set increased. The method presented in this paper controls the tail area-based false discovery rate to distinguish between barcoded and unbarcoded reads. This method helps to establish the highest acceptable minimal distance between reads and barcode sequences. In a proof of concept experiment we correctly detected barcodes in 83% of the reads with a precision of 89%. Sensitivity improved to 99% at 99% precision when the adjacent primer sequence was incorporated in the analysis. The analysis was further improved using a paired end strategy. Following an analysis of the data for sequence variants induced in the Atp1a1 gene of C57BL/6 murine melanocytes by ultraviolet light and conferring resistance to ouabain, we found no evidence of cross-contamination of DNA material between samples. Our method offers a proper quantitative treatment of the problem of detecting barcoded reads in a noisy sequencing environment. It is based on the false discovery rate statistics that allows a proper trade-off between sensitivity and precision to be chosen.


Surgical treatment of brain tumors in infants younger than six months of age and review of the literature.
Brain tumors are rare in infants who are younger than six months of age. These tumors can be challenging to treat surgically. We analyzed a modern series of patients treated by a multidisciplinary team at a tertiary care center and performed a literature review of this unique population. Retrospective clinical data were collected for patients surgically treated for intracranial mass lesions at The Children's Hospital of Philadelphia from 1998 to 2007. Dermoid cysts and other skull-based lesions were excluded from the analysis. Sixteen patients younger than six months of age underwent surgery for primary intracranial mass lesions. The median age of the patients at surgery was 5.2 months (range, 1.4-6 months of age). Children most often presented with a bulging fontanelle, hydrocephalus, or macrocephaly (seven patients). Vomiting was seen in five patients, cranial nerve palsies in one patient, and seizures in three patients. All patients had tumor resections and postoperatively were monitored in the intensive care unit. The final pathology consisted of atypical teratoid/rhabdoid tumor (three patients), primitive neuroectodermal tumor/medulloblastoma (three patients), choroid plexus papilloma (two patients), astrocytoma (two patients), ganglioglioma (two patients), desmoplastic infantile ganglioglioma (two patients), glioblastoma multiforme (one patient), and choroid plexus carcinoma (one patient). Two intraoperative deaths occurred. Of the surviving 14, a gross total resection was achieved in four. Adjuvant therapy was determined by a multidisciplinary team composed of neuro-oncology, neurosurgery, and radiation oncology. Seven patients were treated with chemotherapy, and one patient had proton beam therapy. Five-year overall survival was 45%. The eight surviving patients had neurological sequelae, and developmental outcome was variable. Brain tumors are uncommon in children younger than six months of age. Patients present with a variety of tumor pathologies. Children who survive have neurological sequelae. More studies are necessary to understand the impact that different treatment options, tumor pathology, and tumor location have on neurological outcome.


Contribution of semen to early embryo development: fertilization and beyond.
It has long been thought that the factors affecting embryo and foetal development were exclusively maternally derived; hence, if issues regarding fertility and embryo development were to arise, the blame has traditionally been placed solely on the mother. An escalating interest in how paternal factors influence embryo development, however, has begun to prove otherwise. Evidence suggests that both seminal plasma (SP) and sperm contribute multiple factors that shape embryogenesis. This review thus focuses on the role that semen has in driving early embryonic development, and describes how paternal factors, such as SP, sperm centriole, sperm proteins, sperm RNA, sperm DNA, and its integrity, together with epigenetics, may influence the female reproductive tract and post-fertilization events. The important contributions of paternal factors to embryo development highlight the imperative need for further research in this area, which is sure to bring forth breakthroughs leading to improvements in infertility diagnosis and ART as well as reducing the risk of miscarriage. This review provides a comprehensive overview of the role of human semen in development of the early embryo, with the aim of providing a better understanding of the influence of SP and sperm on early embryonic divisions, gene and protein expression, miscarriage, and congenital diseases. PubMed searches were performed using the terms 'sperm structure', 'capacitation', 'acrosome reaction', 'fertilization', 'oocyte activation', 'PLCζ', 'PAWP', 'sperm-borne oocyte activation factor', 'oocyte activation deficiency', 'sperm centriole', 'sperm transport', 'sperm mitochondria', 'seminal plasma', 'sperm epigenetics', 'sperm histone modifications', 'sperm DNA methylation', 'sperm-derived transcripts', 'sperm-derived proteins', 'sperm DNA fragmentation', 'sperm mRNA', 'sperm miRNAs', 'sperm piRNAs', and 'sperm-derived aneuploidy'. The reviewed articles were restricted to those published in English between 1980 and 2022. The data suggest that male-derived factors contribute much more than just the male haploid genome to the early embryo. Evidence indicates that semen contributes multiple factors that help shape the fate of embryogenesis. These male-derived factors include contributions from SP, the paternal centriole, RNA and proteins, and DNA integrity. In addition, epigenetic changes have an impact on the female reproductive tract, fertilization, and early stages of embryo development. For example, recent proteomic and transcriptomic studies have identified several sperm-borne markers that play important roles in oocyte fertilization and embryogenesis. This review highlights that several male-derived factors are required to work in tandem with female counterparts to allow for correct fertilization and development of the early embryo. A deeper understanding of the contributions of paternal factors that are shuttled over from the sperm cell to the embryo can shed light on how to improve ART from an andrological perspective. Further studies may aid in preventing the passing on of genetic and epigenetic abnormalities of paternal origin, thus decreasing the incidence of male factor infertility. In addition, understanding the exact mechanisms of paternal contribution may assist reproductive scientists and IVF clinicians in determining new causes of recurrent early miscarriage or fertilization failure.


Life after SUDEP: Experiences of traumatic loss and growth.
To understand the experiences of bereaved relatives of individuals who passed due to sudden unexpected death in epilepsy (SUDEP) and to explore the impacts of death in their lives. The principles of fundamental qualitative description informed all design decisions. Stratified purposeful sampling included 21 bereaved relatives (parent, sibling, or spouse/partner), aged at least 18 years, of persons who passed away because of SUDEP. In-depth one-to-one interviews were conducted. Directed content analysis was used to code, categorize, and synthesize the interview data. There was some criticism of emergency response and medical professionals involved in providing insensitive or poor care immediately after SUDEP occurred. Personal hardships described by participants following SUDEP included loss of personal identity, feeling depressed, experiencing guilt, having panic attacks, requiring therapy, as well as having difficulty with anniversaries, dates, and cleaning up a child's room. Bereaved spouses and parents in particular spoke of experiencing challenges in maintaining other relationships following the death. Some participants spoke of experiencing increased financial hardships. Ways of coping included keeping oneself busy, honoring the memory of the loved one, relying on friends and families, and engaging in advocacy/community work, including raising awareness on epilepsy and SUDEP. Sudden unexpected death in epilepsy affected several aspects of the day-to-day lives of bereaved relatives. Though methods of coping were similar to the usual strategies adopted by all bereaved relatives, advocacy work related to raising awareness about epilepsy and SUDEP was unique to this group. Guidelines on SUDEP should ideally include recommendations for trauma-informed support and assessment for depression and anxiety to the bereaved relatives as well.


Correlates of hypertension among urban Asian Indian adolescents.
Primary hypertension has become increasingly common in children but remains largely understudied, underdiagnosed and undertreated. This study examines the relationship between hypertension in adolescents and various markers of obesity, serum lipid levels, fasting blood glucose (FBG), haemoglobin A1c (HbA1c) and family history of hypertension. DESIGN/SETTINGS: A cross-sectional population-based study of 1022 students aged 14-19 years in New Delhi, India. MAIN OUTCOME/EXPOSURES: Those with age, gender and height specific blood pressure >95th percentile (derived from study data) or >130/85 mm Hg were considered hypertensive. Various markers of obesity, FBG, HbA1c and serum lipid levels were divided into quartiles and the odds ratios of hypertension calculated for the highest quartiles with reference to the lowest. Hypertension was seen in 65 (6.4%) adolescents (2.7% isolated systolic, 2.0% isolated diastolic and 1.7% both). The odds of having hypertension were higher for those in the highest versus lowest quartiles of various measures of obesity such as body mass index (OR 2.90; 95% CI 1.40 to 6.12) and waist circumference (OR 5.21; 95% CI 2.14 to 12.17). A parental history of hypertension was associated with diastolic hypertension in the child (OR 2.21; 95% CI 1.13 to 4.33); the odds ratio decreased after simultaneous adjustment for salt intake (OR 1.98; 95% CI 1.00 to 3.94). In a multivariable model with backward elimination, waist circumference and triglycerides were the strongest predictors of hypertension, further suggesting that the relationship is stronger with central than peripheral obesity. Hypertension in Asian Indian adolescents is associated with obesity, higher serum lipids and a family history of hypertension.


K-ATP channels in dopamine substantia nigra neurons control bursting and novelty-induced exploration.
Phasic activation of the dopamine (DA) midbrain system in response to unexpected reward or novelty is critical for adaptive behavioral strategies. This activation of DA midbrain neurons occurs via a synaptically triggered switch from low-frequency background spiking to transient high-frequency burst firing. We found that, in medial DA neurons of the substantia nigra (SN), activity of ATP-sensitive potassium (K-ATP) channels enabled NMDA-mediated bursting in vitro as well as spontaneous in vivo burst firing in anesthetized mice. Cell-selective silencing of K-ATP channel activity in medial SN DA neurons revealed that their K-ATP channel-gated burst firing was crucial for novelty-dependent exploratory behavior. We also detected a transcriptional upregulation of K-ATP channel and NMDA receptor subunits, as well as high in vivo burst firing, in surviving SN DA neurons from Parkinson's disease patients, suggesting that burst-gating K-ATP channel function in DA neurons affects phenotypes in both disease and health.


Can hematological parameters in type 2 diabetes predict microvascular complication development?
To examine potential associations between neutrophil/lymphocyte ratio, platelet/lymphocyte ratio, mean platelet volume (MPV), HbA1c and microvascular complications in diabetic patients from a cost-effectiveness perspective. One hundred patients with type 2 diabetes attending our outpatient unit between May 2018 and October 2018 were included, and 100 healthy individuals served as the control group. A retrospective file search was performed to collect information on hemoglobin, mean platelet volume (MPV), glycosylated haemoglobin (HbA1c), hematocrit (Hct), neutrophil and lymphocyte count, neutrophil/lymphocyte ratio (NLR), platelets (Plt), platelet/lymphocyte ratio (PLR), and microvascular complications (neuropathy, retinopathy, nephropathy). Demographic and laboratory data were retrospectively controlled between diabetes (n=100) and healthy control (n=100) groups. The mean age in diabetic patients and healthy controls was 56.34 and 36.68 years, respectively. The mean NLR in diabetics and healthy controls was 2.48 and 2.11, the difference in NLR being significant (p=0.002). MPV in diabetics and controls was 8.54 and 8.53, respectively, and the difference was not significant (p=0.93). PLR was also similar, i.e. 149.7 and 145.3 in diabetics and healthy controls (p=0.067). With respect to microvascular complications, retinopathy was found to be significantly associated with MPV and NLR (p=0.015, and p=0.051), and nephropathy showed a significant association with NLR (p=0.027) among diabetics. In contrast with the two other microvascular complications, no significant association between neuropathy and NLR could be detected, while PLR and neuropathy was significantly associated (p=0.003). Microvascular complications may be associated with certain hematologic parameters, as suggested by comparisons both between diabetics and healthy individuals and within the group of diabetic individuals. We believe that hematologic parameters such as hematocrit, MPV, NLR, and PLR, which can be obtained through a simple complete blood count, may be utilized as cost-effective predictors of diabetic microvascular complications. Further prospective studies with larger sample size are required to better delineate these associations.


The Worldwide Invasion of Drosophila suzukii Is Accompanied by a Large Increase of Transposable Element Load and a Small Number of Putatively Adaptive Insertions.
Transposable elements (TEs) are ubiquitous and mobile repeated sequences. They are major determinants of host fitness. Here, we characterized the TE content of the spotted wing fly Drosophila suzukii. Using a recently improved genome assembly, we reconstructed TE sequences de novo and found that TEs occupy 47% of the genome and are mostly located in gene-poor regions. The majority of TE insertions segregate at low frequencies, indicating a recent and probably ongoing TE activity. To explore TE dynamics in the context of biological invasions, we studied the variation of TE abundance in genomic data from 16 invasive and six native populations of D. suzukii. We found a large increase of the TE load in invasive populations correlated with a reduced Watterson estimate of genetic diversity θw^ a proxy of effective population size. We did not find any correlation between TE contents and bioclimatic variables, indicating a minor effect of environmentally induced TE activity. A genome-wide association study revealed that ca. 2,000 genomic regions are associated with TE abundance. We did not find, however, any evidence in such regions of an enrichment for genes known to interact with TE activity (e.g., transcription factor encoding genes or genes of the piRNA pathway). Finally, the study of TE insertion frequencies revealed 15 putatively adaptive TE insertions, six of them being likely associated with the recent invasion history of the species.


High-resolution measurement of long-range distances in RNA: pulse EPR spectroscopy with TEMPO-labeled nucleotides.
Structural information at atomic resolution of biomolecular assemblies, such as RNA and RNA protein complexes, is fundamental to comprehend biological function. Modern spectroscopic methods offer exceptional opportunities in this direction. Here we present the capability of pulse EPR to report high-resolution long-range distances in RNAs by means of a recently developed spin labeled nucleotide, which carries the TEMPO group directly attached to the nucleobase and preserves Watson-Crick base-pairing. In a representative RNA duplex with spin-label separations up to 28 base pairs (≈8 nm) we demonstrate that the label allows for a model-free conversion of inter-spin distances into base-pair separation (Δbp) if broad-band pulse excitation at Q band frequencies (34 GHz) is applied. The observed distance distribution increases from ±0.2 nm for Δbp = 10 to only ±0.5 nm for Δbp = 28, consistent with only small deviations from the "ideal" A-form RNA structure. Molecular dynamics (MD) simulations conducted at 20 °C show restricted conformational freedom of the label. MD-generated structural deviations from an "ideal" A-RNA geometry help disentangle the contributions of local flexibility of the label and its neighboring nucleobases and global deformations of the RNA double helix to the experimental distance distributions. The study demonstrates that our simple but strategic spin labeling procedure can access detailed structural information on RNAs at atomic resolution over distances that match the size of macromolecular RNA complexes.


Facilitated cross-species transmission of prions in extraneural tissue.
Prions are infectious pathogens essentially composed of PrP(Sc), an abnormally folded form of the host-encoded prion protein PrP(C). Constrained steric interactions between PrP(Sc) and PrP(C) are thought to provide prions with species specificity and to control cross-species transmission into other host populations, including humans. We compared the ability of brain and lymphoid tissues from ovine and human PrP transgenic mice to replicate foreign, inefficiently transmitted prions. Lymphoid tissue was consistently more permissive than the brain to prions such as those causing chronic wasting disease and bovine spongiform encephalopathy. Furthermore, when the transmission barrier was overcome through strain shifting in the brain, a distinct agent propagated in the spleen, which retained the ability to infect the original host. Thus, prion cross-species transmission efficacy can exhibit a marked tissue dependence.


Metastasis-inducing proteins are widely expressed in human brain metastases and associated with intracranial progression and radiation response.
Understanding the factors that drive recurrence and radiosensitivity in brain metastases would improve prediction of outcomes, treatment planning and development of therapeutics. We investigated the expression of known metastasis-inducing proteins in human brain metastases. Immunohistochemistry on metastases removed at neurosurgery from 138 patients to determine the degree and pattern of expression of the proteins S100A4, S100P, AGR2, osteopontin (OPN) and the DNA repair marker FANCD2. Validation of significant findings in a separate prospective series with the investigation of intra-tumoral heterogeneity using image-guided sampling. Assessment of S100A4 expression in brain metastatic and non-metastatic primary breast carcinomas. There was widespread staining for OPN, S100A4, S100P and AGR2 in human brain metastases. Positive staining for S100A4 was independently associated with a shorter time to intracranial progression after resection in multivariate analysis (hazard ratio for negative over positive staining=0.17, 95% CI: 0.04-0.74, P=0.018). S100A4 was expressed at the leading edge of brain metastases in image guided sampling and overexpressed in brain metastatic vs non-brain metastatic primary breast carcinomas. Staining for OPN was associated with a significant increase in survival time after post-operative whole-brain radiotherapy in retrospective (OPN negative 3.43 months, 95% CI: 1.36-5.51 vs OPN positive, 11.20 months 95% CI: 7.68-14.72, Log rank test, P<0.001) and validation populations. Proteins known to be involved in cellular adhesion and migration in vitro, and metastasis in vivo are significantly expressed in human brain metastases and may be useful biomarkers of intracranial progression and radiosensitivity.


Iron Deficiency Anemia in Pregnancy.
Anemia is a common problem in obstetrics and perinatal care. Any hemoglobin below 10.5 g/dL can be regarded as true anemia regardless of gestational age. Reasons for anemia in pregnancy are mainly nutritional deficiencies, parasitic and bacterial diseases, and inborn red blood cell disorders such as thalassemias. The main cause of anemia in obstetrics is iron deficiency, which has a worldwide prevalence between estimated 20%-80% and consists of a primarily female population. Stages of iron deficiency are depletion of iron stores, iron-deficient erythropoiesis without anemia, and iron deficiency anemia, the most pronounced form of iron deficiency. Pregnancy anemia can be aggravated by various conditions such as uterine or placental bleedings, gastrointestinal bleedings, and peripartum blood loss. In addition to the general consequences of anemia, there are specific risks during pregnancy for the mother and the fetus such as intrauterine growth retardation, prematurity, feto-placental miss ratio, and higher risk for peripartum blood transfusion. Besides the importance of prophylaxis of iron deficiency, the main therapy options for the treatment of pregnancy anemia are oral iron and intravenous iron preparations.


Development of malignant germ cells - the genvironmental hypothesis.
Human germ cell tumors are of interest because of their epidemiology, clinic and patho-biology. Histologically, they are subdivided into various elements, with similarities to embryogenesis. Recent insight triggered development of a higher order division into five types of human germ cell tumors. In the context of male germ cells, only three are relevant; Type I: teratomas and yolk sac tumors of neonates and infants; Type II: seminomas and nonseminomas of (predominantly) adolescents and adults; and Type III: spermatocytic seminomas of the elderly. Various animal models, both occurring spontaneous or induced, are reported, of which their relevance is still a matter of debate. Recent multidisciplinary studies have led to a significant increase in our understanding of the parameters involved in the earliest pathogenetic steps of human germ cells tumors, particularly the seminomas and nonseminomas (Type II). This paper will discuss a number of interesting insights into the normal and aberrant regulation of germ cell development, resulting in the so-called genvironmental hypothesis. This assumes a subtle interaction between environmental- and (epi)genetic parameters, resulting in clinical/phenotypical characteristics. These influence signaling pathways and thereby developmental processes, including gonadal development, germ cell proliferation, maturation and apoptosis. In the case of a disturbed physiology, either due to the germ cell itself, or the micro-environment, embryonic germ cells, during a specific window of sensitization, might be blocked in their maturation, resulting in carcinoma in situ or gonadoblastoma, the precursors of seminomas and nonseminomas. The level of testicularization of the gonad determines the histological composition of the precursor. These insights will allow a better definition of individuals at risk of developing a germ cell malignancy, and allow a better selection of scientific approaches to elucidate the corresponding pathogenesis.


Interleukin-1 gene cluster variants with innate cytokine production profiles and osteoarthritis in subjects from the Genetics, Osteoarthritis and Progression Study.
To assess whether genetic variation in the interleukin-1 (IL-1) gene cluster contributes to familial osteoarthritis (OA) by influencing innate ex vivo production of IL-1beta or IL-1 receptor antagonist (IL-1Ra). Innate ex vivo IL-1beta and IL-1Ra production upon lipopolysaccharide (LPS) stimulation of whole blood cells was measured in subjects from the Genetics, Osteoarthritis and Progression (GARP) Study, which includes sibling pairs in which at least one sibling has symptomatic OA at multiple sites. Radiographic OA (ROA) was assessed by Kellgren/Lawrence score. Subjects from the GARP Study and controls from the Rotterdam Study were genotyped for 7 single-nucleotide polymorphisms (SNPs) encompassing the IL-1 gene cluster on chromosome 2q13. Linkage disequilibrium analysis and genotype and haplotype association analysis were performed to assess the relationship between the IL-1 gene cluster SNPs, innate ex vivo cytokine production, and OA. Among subjects in the GARP Study, the haplotype variable-number tandem repeat in intron 2/T+8006C/T+11100C 2/2/1 of the IL1RN gene was significantly associated with reduced innate ex vivo bioavailability of IL-1beta upon LPS stimulation (P = 0.026) and with ROA at the highest number of joint locations. These results show that genetic variation at the IL-1 gene cluster is associated with lower IL-1beta bioavailability and with OA at a large number of joint locations. The data further indicate that, among subjects with OA affecting the highest number of joints, the innate immune system may be activated, thereby obscuring possible underlying mechanisms.


Genome evolution reveals biochemical networks and functional modules.
The analysis of completely sequenced genomes uncovers an astonishing variability between species in terms of gene content and order. During genome history, the genes are frequently rear-ranged, duplicated, lost, or transferred horizontally between genomes. These events appear to be stochastic, yet they are under selective constraints resulting from the functional interactions between genes. These genomic constraints form the basis for a variety of techniques that employ systematic genome comparisons to predict functional associations among genes. The most powerful techniques to date are based on conserved gene neighborhood, gene fusion events, and common phylogenetic distributions of gene families. Here we show that these techniques, if integrated quantitatively and applied to a sufficiently large number of genomes, have reached a resolution which allows the characterization of function at a higher level than that of the individual gene: global modularity becomes detectable in a functional protein network. In Escherichia coli, the predicted modules can be bench-marked by comparison to known metabolic pathways. We found as many as 74% of the known metabolic enzymes clustering together in modules, with an average pathway specificity of at least 84%. The modules extend beyond metabolism, and have led to hundreds of reliable functional predictions both at the protein and pathway level. The results indicate that modularity in protein networks is intrinsically encoded in present-day genomes.


The role of repulsive guidance molecules in the embryonic and adult vertebrate central nervous system.
During the development of the nervous system, outgrowing axons often have to travel long distances to reach their target neurons. In this process, outgrowing neurites tipped with motile growth cones rely on guidance cues present in their local environment. These cues are detected by specific receptors expressed on growth cones and neurites and influence the trajectory of the growing fibres. Neurite growth, guidance, target innervation and synapse formation and maturation are the processes that occur predominantly but not exclusively during embryonic or early post-natal development in vertebrates. As a result, a functional neural network is established, which is usually remarkably stable. However, the stability of the neural network in higher vertebrates comes at an expensive price, i.e. the loss of any significant ability to regenerate injured or damaged neuronal connections in their central nervous system (CNS). Most importantly, neurite growth inhibitors prevent any regenerative growth of injured nerve fibres. Some of these inhibitors are associated with CNS myelin, others are found at the lesion site and in the scar tissue. Traumatic injuries in brain and spinal cord of mammals induce upregulation of embryonic inhibitory or repulsive guidance cues and their receptors on the neurites. An example for embryonic repulsive directional cues re-expressed at lesion sites in both the rat and human CNS is provided with repulsive guidance molecules, a new family of directional guidance cues.


Requirement for interactions between two subpopulations of T cells for helper cell induction in vitro.
The evidence for the interaction of 2 subpopulations of T cells, short-lived cells sensitive to adult thymectomy (T1 cells), and long-lived recirculating cells, sensitive to the action of antilymphocyte serum (T2 cells) in the induction of helper cells is presented. This T-T interaction occurred across a cell-impermeable nucleopore membrane, indicating that it did not depend on cell contact, but was mediated by subcellular factors. There was no genetic restriction on this T-T interaction, if it was performed across a nucleopore membrane. The implications of these results on our concepts of the mechanism of help are discussed.


Imaging pain in patients: is it meaningful?
Neuroimaging methods are widely used by researchers and clinicians interested in better understanding the functioning of the human brain in health and disease. Advances have been made in understanding how nociceptive processing within the healthy human central nervous system generates a conscious perception of pain. The focus has now shifted towards patient-related research, harnessing earlier developments to test specific hypotheses in a broad range of chronic pain disorders. The timing is ideal to assess the utility of data generated from these studies. This review discusses how clinical pain is represented in the human brain as compared with the processing of acute pain in healthy controls. The imaging literature is reviewed for hypotheses that have been tested in patients regarding mechanisms that might contribute towards the development of chronic pain. Issues related to plasticity, central sensitization, psychological confounds, genetics, and necrosis are examined. Results to date strongly support the notion that neuroimaging will aid our understanding of basic mechanisms contributing to the generation of chronic pain states. These techniques might help diagnose a patient's pain condition in a more objective and robust way, enabling better targeting of therapies and rapid development of compounds to alleviate pain.


Health systems' responsiveness and its characteristics: a cross-country comparative analysis.
OBJECTIVES. Responsiveness has been identified as one of the intrinsic goals of health care systems. Little is known, however, about its determinants. Our objective is to investigate the potential country-level drivers of health system responsiveness. DATA SOURCE. Data on responsiveness are taken from the World Health Survey. Information on country-level characteristics is obtained from a variety of sources including the United Nations Development Program (UNDP). STUDY DESIGN. A two-step procedure. First, using survey data we derive a country-level measure of system responsiveness purged of differences in individual reporting behavior. Secondly, we run cross-sectional country-level regressions of responsiveness on potential drivers. PRINCIPAL FINDINGS. Health care expenditures per capita are positively associated with responsiveness, after controlling for the influence of potential confounding factors. Aspects of responsiveness are also associated with public sector spending (negatively) and educational development (positively). CONCLUSIONS. From a policy perspective, improvements in responsiveness may require higher spending levels. The expansion of nonpublic sector provision, perhaps in the form of increased patient choice, may also serve to improve responsiveness. However, these inferences are tentative and require further study.


Safety and efficacy of risdiplam in patients with type 1 spinal muscular atrophy (FIREFISH part 2): secondary analyses from an open-label trial.
Risdiplam is an orally administered therapy that modifies pre-mRNA splicing of the survival of motor neuron 2 (SMN2) gene and is approved for the treatment of spinal muscular atrophy. The FIREFISH study is investigating the safety and efficacy of risdiplam in treated infants with type 1 spinal muscular atrophy versus historical controls. The primary endpoint of part 2 of the FIREFISH study showed that infants with type 1 spinal muscular atrophy attained the ability to sit without support for at least 5 s after 12 months of treatment. Here, we report on the safety and efficacy of risdiplam in FIREFISH part 2 over 24 months of treatment. FIREFISH is an ongoing, multicentre, open-label, two-part study. In FIREFISH part 2, eligible infants (aged 1-7 months at enrolment, with a genetically confirmed diagnosis of spinal muscular atrophy, and two SMN2 gene copies) were enrolled in 14 hospitals in ten countries across Europe, North America, South America, and Asia. Risdiplam was orally administered once daily at 0·2 mg/kg for infants between 5 months and 2 years of age; once an infant reached 2 years of age, the dose was increased to 0·25 mg/kg. Infants younger than 5 months started at 0·04 mg/kg (infants between 1 month and 3 months old) or 0·08 mg/kg (infants between 3 months and 5 months old), and this starting dose was adjusted to 0·2 mg/kg once pharmacokinetic data were available for each infant. The primary and secondary endpoints included in the statistical hierarchy and assessed at month 12 have been reported previously. Here we present the remainder of the secondary efficacy endpoints that were included in the statistical hierarchy at month 24: the ability to sit without support for at least 30 s, to stand alone, and to walk alone, as assessed by the Bayley Scales of Infant and Toddler Development, third edition gross motor subscale. These three endpoints were compared with a performance criterion of 5% that was defined based on the natural history of type 1 spinal muscular atrophy; the results were considered statistically significant if the lower limit of the two-sided 90% CI was above the 5% threshold. FIREFISH is registered with ClinicalTrials.gov, NCT02913482. Recruitment is closed; the 36-month extension period of the study is ongoing. Between March 13 and Nov 19, 2018, 41 infants were enrolled in FIREFISH part 2. After 24 months of treatment, 38 infants were ongoing in the study and 18 infants (44% [90% CI 31-58]) were able to sit without support for at least 30 s (p<0·0001 compared with the performance criterion derived from the natural history of untreated infants with type 1 spinal muscular atrophy). No infants could stand alone (0 [90% CI 0-7]) or walk alone (0 [0-7]) after 24 months of treatment. The most frequently reported adverse event was upper respiratory tract infection, in 22 infants (54%); the most common serious adverse events were pneumonia in 16 infants (39%) and respiratory distress in three infants (7%). Treatment with risdiplam over 24 months resulted in continual improvements in motor function and achievement of developmental motor milestones. The FIREFISH open-label extension phase will provide additional evidence regarding long-term safety and efficacy of risdiplam. F Hoffmann-La Roche.


Acquired FXIII inhibitors: a systematic review.
Coagulation factor XIII (FXIII) is a protein that promotes fibrin stabilization by forming multiple covalent cross-links between fibrin monomers. Beside congenital FXIII deficiency, due to FXIII gene mutations, severe acquired FXIII deficiency has been described in association with autoantibodies against coagulation FXIII. These inhibitors, which occurs very rarely but may cause life-threatening bleeding complications, may arise spontaneously or in association with autoimmune and lymphoproliferative disorders or medications. The management of patients with acquired FXIII inhibitors is very demanding and treatment regimens must be focused on eradication of the inhibitor and to increase the plasma FXIII levels. In this systematic review, we analyse all the published case-reports on anti-FXIII autoantibodies focusing on the clinical features and treatment modalities of this acquired hemorrhagic condition.


Laser induced forward transfer of metals by temporally shaped femtosecond laser pulses.
Temporally shaped, femtosecond laser pulses have been used for controlling the size and the morphology of micron-sized metallic structures obtained by using the Laser Induced Forward Transfer (LIFT) technique. We report the effect of pulse shaping on the size and morphology of the deposited structures of Au, Zn, Cr on a function of the pulse separation time ??t (from 0 to 10 ps) of double pulses of variable intensities generated by using a liquid crystal spatial light modulator (SLM). The observed differences in size and morphology are correlated with the outcome of pump-probe experiments for the study of electron-phonon scattering dynamics and subsequent energy transfer processes to the bulk in the different metals employed. We propose that in metals with weak electron-lattice coupling, the electron ballistic motion and the resulting fast electron scattering at the film surface, as well as the internal electron thermalization process are crucial to the morphology and size of the transferred material. Therefore, temporal shaping within the corresponding time scales of these processes may be used for tailoring the features of the metallic structures obtained by LIFT.


Clinical review of pulmonary disease caused by Mycobacterium xenopi.
Mycobacterium xenopi comprised 56% of all non-tuberculous mycobacteria isolated in the Brompton Hospital laboratory during six years. M xenopi alone was cultured from the sputum of 23 patients, whose clinical and bacteriological features are reviewed. Pulmonary disease was considered to be due to the organism present in 15 of these patients on the basis of strict criteria for a causal relationship. Radiographs of all these patients were consistent with mycobacterial disease, showing pulmonary cavitation in 11, apical shadowing in three, and upper lobe fibrosis in one. In nine patients the disease was chronic, progressing very slowly with longstanding respiratory symptoms and extensive radiographic abnormalities developing over many years (mean 19 years). In six patients the disease was subacute, being of recent onset with radiographic shadowing of limited extent. M xenopi was isolated as a saprophyte from four patients who had lung disease of known cause and single isolates were obtained from four patients who had no lung disease attributable to this organism. In vitro drug sensitivity tests showed the sputum cultures of 17 of 22 patients to be sensitive to two or more of the drugs rifampicin, isoniazid, and ethambutol. Response to chemotherapy was unpredictable and did not always correlate with the results of sensitivity tests. Of 11 patients treated with at least two drugs to which their organisms were sensitive, two remained sputum positive, one relapsed after 18 months of chemotherapy, and eight have remained sputum negative during a minimum of three years' follow-up.


Functional interaction of Src family kinases with the acetylcholine receptor in C2 myotubes.
Tyrosine phosphorylation of the beta subunit of the acetylcholine receptor (AChR) has been postulated to play a role in AChR clustering during development of the neuromuscular junction. We have investigated the mechanism of this phosphorylation in mammalian C2 myotubes and report that the tyrosine kinase Src binds and phosphorylates glutathione S-transferase fusion proteins containing the N-terminal half of the cytoplasmic loop of the beta subunit. No binding occurs to the related kinases Fyn or Yes or to the corresponding regions from the gamma and delta subunits. Furthermore, AChRs affinity-isolated from C2 myotubes using alpha-bungarotoxin-Sepharose were specifically associated with Src and Fyn and had tyrosine-phosphorylated beta subunits. We suggest that AChRs are initially phosphorylated by Src and subsequently bind Fyn in a phosphotyrosine-dependent manner. These interactions are likely to play an important role in construction of the specialized postsynaptic membrane during synaptogenesis.


Effect of platelet-rich plasma on the peri-implant bone response: an experimental study.
Platelet-rich plasma (PRP) is used as a source of growth factors to stimulate and accelerate bone formation and soft tissue healing. The use of PRP in bone regeneration, both around dental implants and in periodontic treatments, has become particularly appealing. The aim of this study was to evaluate the effect of PRP in an experimental model of osteogenesis around laminar implants. Fifteen male Wistar rats, weighing 90 +/- 10 g, were used in this study. One milliliter of blood was obtained from each animal by intracardiac puncture and transferred into Eppendorf tubes containing 10% sodium citrate. The tubes were centrifuged at 1500 rpm for 15 minutes and PRP was prepared. The laminar test was used to evaluate the bone peri-implant response. PRP and a titanium laminar implant were introduced into the right tibia (Ti/PRP group), whereas the left tibia (control) received only a laminar implant (Ti group). Thirty days postimplantation, the tibiae were resected, radiographed, and processed for embedding in acrylic resin. Ground sections (50 microm) were stained with toluidine blue. The peri-implant bone volume was evaluated histomorphometrically. Statistical analysis of the data was performed. The amount of newly formed bone in the Ti/PRP group (30 +/- 7 cm) was significantly greater than in the Ti group (16 +/- 3 cm). A greater volume of peri-implant bone was observed when PRP was used in the laminar implant test model.


Surface-enhanced Raman spectral biomarkers correlate with Ankle Brachial Index and characterize leg muscle biochemical composition of patients with peripheral arterial disease.
Peripheral arterial disease (PAD) is characterized by atherosclerotic blockages of the arteries supplying the lower extremities, which cause a progressive accumulation of ischemic injury to the skeletal muscles of the lower limbs. This injury includes altered metabolic processes, damaged organelles, and compromised bioenergetics in the affected muscles. The objective of this study was to explore the association of Raman spectral signatures of muscle biochemistry with the severity of atherosclerosis in the legs as determined by the Ankle Brachial Index (ABI) and clinical presentation. We collected muscle biopsies from the gastrocnemius (calf muscle) of five patients with clinically diagnosed claudication, five patients with clinically diagnosed critical limb ischemia (CLI), and five control patients who did not have PAD. A partial least squares regression (PLSR) model was able to predict patient ABI with a correlation coefficient of 0.99 during training and a correlation coefficient of 0.85 using a full cross-validation. When using the first three PLS factor scores in combination with linear discriminant analysis, the discriminant model was able to correctly classify the control, claudicating, and CLI patients with 100% accuracy, using a full cross-validation procedure. Raman spectroscopy is capable of detecting and measuring unique biochemical signatures of skeletal muscle. These signatures can discriminate control muscles from PAD muscles and correlate with the ABI and clinical presentation of the PAD patient. Raman spectroscopy provides novel spectral biomarkers that may complement existing methods for diagnosis and monitoring treatment of PAD patients.


The effects of nutrient enrichment and herbivore abundance on the ability of turf algae to overgrow coral in the Caribbean.
Turf algae are multispecies communities of small marine macrophytes that are becoming a dominant component of coral reef communities around the world. To assess the impact of turf algae on corals, we investigated the effects of increased nutrients (eutrophication) on the interaction between the Caribbean coral Montastraea annularis and turf algae at their growth boundary. We also assessed whether herbivores are capable of reducing the abundance of turf algae at coral-algae boundaries. We found that turf algae cause visible (overgrowth) and invisible negative effects (reduced fitness) on neighbouring corals. Corals can overgrow neighbouring turf algae very slowly (at a rate of 0.12 mm 3 wk(-1)) at ambient nutrient concentrations, but turf algae overgrew corals (at a rate of 0.34 mm 3 wk(-1)) when nutrients were experimentally increased. Exclusion of herbivores had no measurable effect on the rate turf algae overgrew corals. We also used PAM fluorometry (a common approach for measuring of a colony's "fitness") to detect the effects of turf algae on the photophysiology of neighboring corals. Turf algae always reduced the effective photochemical efficiency of neighbouring corals, regardless of nutrient and/or herbivore conditions. The findings that herbivores are not capable of controlling the abundance of turf algae and that nutrient enrichment gives turf algae an overall competitive advantage over corals together have serious implications for the health of Caribbean coral reef systems. At ambient nutrient levels, traditional conservation measures aimed at reversing coral-to-algae phase shifts by reducing algal abundance (i.e., increasing herbivore populations by establishing Marine Protected Areas or tightening fishing regulations) will not necessarily reduce the negative impact of turf algae on local coral communities. Because turf algae have become the most abundant benthic group on Curaçao (and likely elsewhere in the Caribbean), new conservation strategies are required to mitigate their negative impact on coral communities.


The metabolic syndrome and cognitive performance: the Northern Manhattan Study.
The metabolic syndrome (MetS) is a risk factor for diabetes, stroke, myocardial infarction, and increased mortality, and has been associated with cognition in some populations. We hypothesized that MetS would be associated with lower Mini-Mental State Examination (MMSE) scores in a multi-ethnic population, and that MetS is a better predictor of cognition than its individual components or diabetes. We conducted a cross-sectional analysis among 3,150 stroke-free participants. MetS was defined by the modified National Cholesterol Education Program guidelines-Adult Treatment Panel III (NCEP-ATPIII) criteria. Linear regression and polytomous logistic regression estimated the association between MMSE score and MetS, its individual components, diabetes, and inflammatory biomarkers. MetS was inversely associated with MMSE score (unadjusted β = -0.67; 95% CI -0.92, -0.41). Adjusting for potential confounders, MetS was associated with lower MMSE score (adjusted β = -0.24; 95% CI -0.47, -0.01), but its individual components and diabetes were not. Those with MetS were more likely to have an MMSE score of <18 than a score of ≥ 24 (adjusted OR = 1.94; 95% CI 1.26, 3.01). There was an interaction between MetS and race-ethnicity, such that MetS was associated with lower MMSE score among non-Hispanic whites and Hispanics but not non-Hispanic blacks. MetS was associated with lower cognition in a multi-ethnic population. Further studies of the effect of MetS on cognition are warranted, and should account for demographic differences.


Comparative In Vitro Evaluation of Antibacterial and Osteogenic Activity of Polysaccharide and Flavonoid Fractions Isolated from the leaves of Saussurea controversa.
The osteogenic, cytotoxic, and antibacterial activities of polysaccharide (PS-SC) and flavonoid (F-SC) fractions isolated from the leaves extract of Saussurea controversa were studied in vitro. F-SC consists of the five quercetin glycosides in the ratio 2:8:10:1:4, which were isolated from the leaves extract of S. controversa and have been characterized previously. PS-SC was first isolated from the leaves extract of S. controversa and has been described. PS-SC consists in 30 compounds is characterized by a high degree of heterogeneity with a heterogeneity index of 19.74. The Mw and Mn of PS-SC were 108.6 and 5.5 kDa, respectively. Structural fragments are represented by galactose, arabinose, xylose, glucose, uronic acids, mannose, and rhamnose in a 10.1:3.3:2.2:2.1:1.7:0.9:0.5 molar ratio. F-SC as compared with PS-SC showed in vitro microbicidal (50 g/L) and better bacteriostatic (6.25 g/L versus 25 g/L of PS-SC) effects against the 24-h growth of Staphylococcus aureus strain 209 P and a 21-day absence of cytotoxicity on human adipose-derived multipotent mesenchymal stromal cells (hAMMSCs). Both fractions (PS-SC>F-SC) at doses of 10-50 mg/L stimulated differentiation of hAMMSCs into secreting osteoblasts accompanied by local mineralization of extracellular matrix. These fractions of S. controversa and especially F-SC, might be promising peroral drugs in the complex treatment of bone fractures and for prophylaxis of their infectious complications.


In vitro performance of lipid-PLGA hybrid nanoparticles as an antigen delivery system: lipid composition matters.
Due to the many beneficial properties combined from both poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) and liposomes, lipid-PLGA hybrid NPs have been intensively studied as cancer drug delivery systems, bio-imaging agent carriers, as well as antigen delivery vehicles. However, the impact of lipid composition on the performance of lipid-PLGA hybrid NPs as a delivery system has not been well investigated. In this study, the influence of lipid composition on the stability of the hybrid NPs and in vitro antigen release from NPs under different conditions was examined. The uptake of hybrid NPs with various surface charges by dendritic cells (DCs) was carefully studied. The results showed that PLGA NPs enveloped by a lipid shell with more positive surface charges could improve the stability of the hybrid NPs, enable better controlled release of antigens encapsulated in PLGA NPs, as well as enhance uptake of NPs by DC.


Lessons on pattern formation from planet WATOR.
It is well known that if reacting species experience unequal diffusion rates, then dynamics that lead to a constant steady state in a "well-mixed" environment can in a spatial setting lead to interesting patterns. In this paper, we focus on complementary pattern formation mechanisms that operate even when the diffusion rates are equal. In particular, we can say that when the mean-field ODE has an attracting periodic orbit then the stochastic spatial model will have large-scale spatial structures in equilibrium. We explore this mechanism in depth through the dynamics of the simulator WATOR.


A Comparison of the Effects of Alpha and Medical-Grade Honey Ointments on Cutaneous Wound Healing in Rats.
Introduction. This study compared the healing efficacy and possible adverse effects of topical Alpha and medical-grade honey ointments on cutaneous wounds in rats. Methods. To conduct the study, 22 male Sprague-Dawley rats were randomly allocated into two equal groups: (1) rats with Alpha ointment applied to the wound surface area and (2) rats with medical-grade honey ointment applied to their wounds. The ointments were applied daily during the 21-day study period. Wound contraction was examined photographically with images taken on days 0, 7, and 21 after wounding. The healing process was histopathologically assessed using skin biopsies taken from the wound sites on days 7 and 21. Results. No statistically significant difference in mean wound surface area was observed between the two study groups. According to histopathological assessment, a significant reduction in the amount of collagen deposition (P value: 0.007) and neovascularisation (P value: 0.002) was seen in the Alpha-treated rats on day 21. No tissue necrosis occurred following the application of Alpha ointment. Conclusion. Daily topical usage of Alpha ointment on a skin wound can negatively affect the healing process by inhibiting neovascularization. Topical Alpha ointment can reduce the possibility of excessive scar formation by reducing collagen deposition.


Clinical biomarkers of pulmonary carcinoid tumors in never smokers via profiling miRNA and target mRNA.
miRNAs play key regulatory roles in cellular pathological processes. We aimed to identify clinically meaningful biomarkers in pulmonary carcinoid tumors (PCTs), a member of neuroendocrine neoplasms, via profiling miRNAs and mRNAs. From the total of 1145 miRNAs, we obtained 16 and 17 miRNAs that showed positive and negative fold changes (FCs, tumors vs. normal tissues) in the top 1% differentially expressed miRNAs, respectively. We uncovered the target genes that were predicted by at least two prediction tools and overlapped by at least one-half of the top miRNAs, which yielded 44 genes (FC<-2) and 56 genes (FC>2), respectively. Higher expressions of CREB5, PTPRB and COL4A3 predicted favorable disease free survival (Hazard ratio: 0.03, 0.19 and 0.36; P value: 0.03, 0.03 and 0.08). Additionally, 79 mutated genes have been found in nine PCTs where TP53 was the only repeated mutation. We identified that the expressions of three genes have clinical implications in PCTs. The biological functions of these biomarkers warrant further studies.


Towards salutogenesis in the development of personalised and preventive healthcare.
The purpose of this review is to discuss how a salutogenetic approach that takes into consideration the human being as physical, psychological and spiritual entity may provide some answers to the difficulties faced by healthcare systems. The choice of medical intervention needs to take into account the technological advances of biomedicine but tailor them to the physical, psychological and spiritual needs of the patient in the context of their biography. Such person-centred medicine aims to strengthen Antonovsky's concepts of resilience and sense of coherence with each therapeutic intervention so that overcoming illness becomes the foundation for better future health. Appropriate evaluation parameters need to be developed and included in order to evaluate the success of interventions in a person-centred, salutogenetic system.


Differential requirement for utrophin in the induced pluripotent stem cell correction of muscle versus fat in muscular dystrophy mice.
Duchenne muscular dystrophy (DMD) is an incurable degenerative muscle disorder. We injected WT mouse induced pluripotent stem cells (iPSCs) into mdx and mdx∶utrophin mutant blastocysts, which are predisposed to develop DMD with an increasing degree of severity (mdx <<< mdx∶utrophin). In mdx chimeras, iPSC-dystrophin was supplied to the muscle sarcolemma to effect corrections at morphological and functional levels. Dystrobrevin was observed in dystrophin-positive and, at a lesser extent, utrophin-positive areas. In the mdx∶utrophin mutant chimeras, although iPSC-dystrophin was also supplied to the muscle sarcolemma, mice still displayed poor skeletal muscle histopathology, and negligible levels of dystrobrevin in dystrophin- and utrophin-negative areas. Not only dystrophin-expressing tissues are affected by iPSCs. Mdx and mdx∶utrophin mice have reduced fat/body weight ratio, but iPSC injection normalized this parameter in both mdx and mdx∶utrophin chimeras, despite the fact that utrophin was compromised in the mdx∶utrophin chimeric fat. The results suggest that the presence of utrophin is required for the iPSC-corrections in skeletal muscle. Furthermore, the results highlight a potential (utrophin-independent) non-cell autonomous role for iPSC-dystrophin in the corrections of non-muscle tissue like fat, which is intimately related to the muscle.


Monoclonal antibodies directed to fucoidan preparations from brown algae.
Cell walls of the brown algae contain a diverse range of polysaccharides with useful bioactivities. The precise structures of the sulfated fucan/fucoidan group of polysaccharides and their roles in generating cell wall architectures and cell properties are not known in detail. Four rat monoclonal antibodies, BAM1 to BAM4, directed to sulfated fucan preparations, have been generated and used to dissect the heterogeneity of brown algal cell wall polysaccharides. BAM1 and BAM4, respectively, bind to a non-sulfated epitope and a sulfated epitope present in the sulfated fucan preparations. BAM2 and BAM3 identified additional distinct epitopes present in the fucoidan preparations. All four epitopes, not yet fully characterised, occur widely within the major brown algal taxonomic groups and show divergent distribution patterns in tissues. The analysis of cell wall extractions and fluorescence imaging reveal differences in the occurrence of the BAM1 to BAM4 epitopes in various tissues of Fucus vesiculosus. In Ectocarpus subulatus, a species closely related to the brown algal model Ectocarpus siliculosus, the BAM4 sulfated epitope was modulated in relation to salinity levels. This new set of monoclonal antibodies will be useful for the dissection of the highly complex and yet poorly resolved sulfated polysaccharides in the brown algae in relation to their ecological and economic significance.


Advanced modelling of commuter choice model and work from home during COVID-19 restrictions in Australia.
The decision to work from home (WFH) or to commute during COVID-19 is having a major structural impact on individuals' travel, work and lifestyle. There are many possible factors influencing this non-marginal change, some of which are captured by objective variables while others are best represented by a number of underlying latent traits captured by attitudes towards WFH and the use of specific modes of transport for the commute that have a bio-security risk such as public transport (PT). We develop and implement a hybrid choice model to investigate the sources of influence, accounting for the endogenous nature of latent soft variables for workers in metropolitan areas in New South Wales and Queensland. The data was collected between September-October 2020, during a period of no lockdown and relatively minor restrictions on workplaces and public gatherings. The results show that one of the most important attributes defining the WFH loving attitude is the workplace policy towards WFH, with workers that can decide where to work having a higher probability of WFH, followed by those that are being directed to, relative to other workplace policies. The bio-security concern with using shared modes such as public transport is a key driver of WFH and choosing to commute via the safer environment of the private car.


Liquid chromatography-mass spectrometry calibration transfer and metabolomics data fusion.
Metabolic profiling is routinely performed on multiple analytical platforms to increase the coverage of detected metabolites, and it is often necessary to distribute biological and clinical samples from a study between instruments of the same type to share the workload between different laboratories. The ability to combine metabolomics data arising from different sources is therefore of great interest, particularly for large-scale or long-term studies, where samples must be analyzed in separate blocks. This is not a trivial task, however, due to differing data structures, temporal variability, and instrumental drift. In this study, we employed blood serum and plasma samples collected from 29 subjects diagnosed with small cell lung cancer and analyzed each sample on two liquid chromatography-mass spectrometry (LC-MS) platforms. We describe a method for mapping retention times and matching metabolite features between platforms and approaches for fusing data acquired from both instruments. Calibration transfer models were developed and shown to be successful at mapping the response of one LC-MS instrument to another (Procrustes dissimilarity = 0.04; Mantel correlation = 0.95), allowing us to merge the data from different samples analyzed on different instruments. Data fusion was assessed in a clinical context by comparing the correlation of each metabolite with subject survival time in both the original and fused data sets: a simple autoscaling procedure (Pearson's R = 0.99) was found to improve upon a calibration transfer method based on partial least-squares regression (R = 0.94).


Pulmonary aspiration in adjustable gastric banding carriers undergoing a second surgical procedure. Considerations on personal experience and review of the literature.
The observation of a relatively high number of pulmonary aspirations (PA) among gastric band (GB) carriers undergoing a second surgery, prompted us to modify our strategy for GB patients candidate to further operation under general anesthesia. In January 2013, following the occurrence of PA at the induction of general anesthesia in 1 GB carrier undergoing a further operation, we reviewed our Data Base between January 2005 and 2013, to explore the rate of pulmonary aspiration in patients GB carriers undergoing a second surgery. Considering the rate (3/172 - 1.7%) too high in comparison with non-GB carriers, we decided to deflate the banding before any further surgery planned under general anesthesia. We then retrospectively reviewed the occurrence of PA after having changed the protocol. Since February 2013, through December 2016, 81 GB carriers underwent a second surgery and not a single episode of PA occurred (0/81). The occurrence of PA in patients with GB seems greater than in non-GB patients. Larger series should be examined to assess the incidence of PA among this specific population. Awareness of the increased risk is important to general anesthesiologists and surgeons, considering the increasing number of GB carriers who may be in need of surgery. Our result after adopting the deflation policy, even though not statistically significant, seems highly suggestive. We believe that, considering the potentially severe consequences of PA, the gastric band should be deflated before any planned procedure requiring general anesthesia. Further data are needed. Adjustable gastric banding, Aspiration Pneumonia, Bariatric surgery, Morbid obesity, Pulmonary Aspiration.


Determining medical decision-making capacity in brain tumor patients: why and how?
Brain tumor patients are at high risk of impaired medical decision-making capacity (MDC), which can be ethically challenging because it limits their ability to give informed consent to medical treatments or participation in research. The European Association of Neuro-Oncology Palliative Care Multidisciplinary Task Force performed a systematic review to identify relevant evidence with respect to MDC that could be used to give recommendations on how to cope with reduced MDC in brain tumor patients. A literature search in several electronic databases was conducted up to September 2019, including studies with brain tumor and other neurological patients. Information related to the following topics was extracted: tools to measure MDC, consent to treatment or research, predictive patient- and treatment-related factors, surrogate decision making, and interventions to improve MDC. A total of 138 articles were deemed eligible. Several structured capacity-assessment instruments are available to aid clinical decision making. These instruments revealed a high incidence of impaired MDC both in brain tumors and other neurological diseases for treatment- and research-related decisions. Incapacity appeared to be mostly determined by the level of cognitive impairment. Surrogate decision making should be considered in case a patient lacks capacity, ensuring that the patient's "best interests" and wishes are guaranteed. Several methods are available that may help to enhance patients' consent capacity. Clinical recommendations on how to detect and manage reduced MDC in brain tumor patients were formulated, reflecting among others the timing of MDC assessments, methods to enhance patients' consent capacity, and alternative procedures, including surrogate consent.


Practical and predictive bioinformatics methods for the identification of potentially cross-reactive protein matches.
A bioinformatics comparison of proteins introduced into food crops through genetic engineering provides a mechanism to identify those proteins that may present an increased risk of allergic reactions for individuals with existing allergies. The goal is to identify proteins that are known to be allergens or are so similar to an allergen that they may induce allergic cross-reactions. Three comparative approaches have traditionally been used, or considered for safety evaluations. One identifies any short (6-8) amino acid segment of the protein that exactly matches a known allergen sequence. The second is an overall primary sequence comparison using Basic Local Alignment Search Tool (BLAST) or FASTA to find matches of greater than 35% identity over 80 amino acids. The third is based on 3-D prediction programs to identify 3-D similarities that might predict potential cross-reactivity. The utility of each of these approaches was debated in the bioinformatics workshop. The consensus agreement from the expert workshop participants was that the short-segment match (e. g., 6-8 amino acids) provides an unacceptably high rate of false positive matches and an uncertain rate of true positive matches, and was not particularly useful for an allergenicity evaluation performed in the context of comprehensive safety evaluation. There was no consensus regarding the most appropriate bioinformatics method, an acceptable scoring criteria for triggering closer examination subsequent to a positive match, or an acceptable scoring mechanism for ranking the utility of the various 3-D approaches that were discussed during the workshop. However, the general consensus was that the most practical approach at this time is to evaluate primary sequence identities to known allergens using either FASTA or BLAST. While there was good agreement that identities of greater than 35% over 80 or more amino acids (recommended by Codex in 2003) is quite conservative, the conclusion was that additional data or studies would be needed to justify changing this criterion as there is some evidence that some individuals sensitized to proteins in evolutionarily conserved protein families may experience cross-reactions to proteins sharing approximately 40% identity.


Correlation of serum hepcidin levels with disease progression in hepatitis B virus-related disease assessed by nanopore film based assay.
Chronic hepatitis B virus (HBV) infection often develop into cirrhosis, and both are major risk factors of hepatocellular carcinoma. However, effective approaches for the monitoring of HBV-related disease progress are still in need. Increased iron storage has an important role in HBV-related diseases. Hepcidin is a key regulator of iron homeostasis whose expression changes are often indicative of abnormal iron metabolism. There are few reports of hepcidin levels in patients with HBV infections, and the available results are inconsistent. In this study, using a recently validated nanopore silica film based method, we measured serum hepcidin levels in 46 HBV-related patients and 20 healthy controls. Patients were divided into three groups: chronic hepatitis B without cirrhosis; HBV-related cirrhosis; and HBV-related cirrhosis with hepatocellular carcinoma. Compared to healthy controls, the mean serum hepcidin level was significantly higher in CHB patients without cirrhosis, and in those with hepatocellular carcinoma, but not in those with cirrhosis. Iron-loading, viral infection and liver dysfunction are determined to be the major regulators of hepcidin in these patients. These observations suggest correlations between serum hepcidin and progression of chronic HBV infection, and may shed a new light on the development of biomarkers for HBV-related disease surveillance.


Growth differentiation factor 15 is a myomitokine governing systemic energy homeostasis.
Reduced mitochondrial electron transport chain activity promotes longevity and improves energy homeostasis via cell-autonomous and -non-autonomous factors in multiple model systems. This mitohormetic effect is thought to involve the mitochondrial unfolded protein response (UPRmt), an adaptive stress-response pathway activated by mitochondrial proteotoxic stress. Using mice with skeletal muscle-specific deficiency of Crif1 (muscle-specific knockout [MKO]), an integral protein of the large mitoribosomal subunit (39S), we identified growth differentiation factor 15 (GDF15) as a UPRmt-associated cell-non-autonomous myomitokine that regulates systemic energy homeostasis. MKO mice were protected against obesity and sensitized to insulin, an effect associated with elevated GDF15 secretion after UPRmt activation. In ob/ob mice, administration of recombinant GDF15 decreased body weight and improved insulin sensitivity, which was attributed to elevated oxidative metabolism and lipid mobilization in the liver, muscle, and adipose tissue. Thus, GDF15 is a potent mitohormetic signal that safeguards against the onset of obesity and insulin resistance.


An echistatin C-terminal peptide activates GPIIbIIIa binding to fibrinogen, fibronectin, vitronectin and collagen type I and type IV.
Integrin binding to proteins often involves recognition of domains containing the arginine-glycine-aspartate (RGD) motif. Different binding affinities and specificities of the integrin-ligand protein interactions involve additional protein domains. The n.m.r. structure of the snake-venom protein echistatin suggested that the C-terminal portion of the molecule might be important, in addition to the RGD domain, in binding to the integrin glycoprotein IIbIIIa (GPIIbIIIa) [Saudek, Atkinson and Pelton (1991) Biochem. 30, 7369-7372]. The synthetic C-terminal peptide, echistatin-(40-49), PRNPHKGPAT, (1) inhibited binding of GPIIbIIIa to immobilized echistatin (IC50 3-6 mM), but did not inhibit binding of GPIIbIIIa to immobilized fibrinogen (up to 5 mM peptide), (2) activated GPIIbIIIa binding to fibronectin and vitronectin, usual ligands for the activated integrin, (3) activated binding of GPIIbIIIa to collagen type I and type IV, proteins not usually regarded as ligands for the integrin, and (4) stimulated 125I-fibrinogen binding by human platelets. These findings argue for an interaction of this non-RGD domain in echistatin with GPIIbIIIa, leading to activation of the integrin and extension of the ligand specificity to include immobilized collagen.


Dynamics of molecular evolution in RNA virus populations depend on sudden versus gradual environmental change.
Understanding the dynamics of molecular adaptation is a fundamental goal of evolutionary biology. While adaptation to constant environments has been well characterized, the effects of environmental complexity remain seldom studied. One simple but understudied factor is the rate of environmental change. Here we used experimental evolution with RNA viruses to investigate whether evolutionary dynamics varied based on the rate of environmental turnover. We used whole-genome next-generation sequencing to characterize evolutionary dynamics in virus populations adapting to a sudden versus gradual shift onto a novel host cell type. In support of theoretical models, we found that when populations evolved in response to a sudden environmental change, mutations of large beneficial effect tended to fix early, followed by mutations of smaller beneficial effect; as predicted, this pattern broke down in response to a gradual environmental change. Early mutational steps were highly parallel across replicate populations in both treatments. The fixation of single mutations was less common than sweeps of associated "cohorts" of mutations, and this pattern intensified when the environment changed gradually. Additionally, clonal interference appeared stronger in response to a gradual change. Our results suggest that the rate of environmental change is an important determinant of evolutionary dynamics in asexual populations.


Practical considerations for the rapid screening for pesticides using ambient pressure desorption ionisation with high-resolution mass spectrometry.
A rapid screening method for pesticides has been developed to streamline the processing of produce entering the United States. Foam swabs were used to recover multi-class mixtures of 240, 140, 132 and 60 pesticides from the surfaces of apples, kiwis, peaches and tomatoes. The mixtures were selected to span a large range of chemical classes, polarities, solubilities and sizes to provide a broad look at how this technique will perform for a variety of analytes. The swabs were analysed using direct analysis in real-time (DART) ionisation coupled with a high-resolution Exactive Orbitrap™ mass spectrometer. This study expands the types of commodities analysed using this method and explores the feasibility of compositing multiple units of produce per batch to analyse a representative sample. It was established that whilst smooth-skinned produce, such as apples, maintained a high detection rate for the pesticide mixtures even when ten apples are swabbed with one foam disk, commodities with rougher surfaces, such as peaches, suffered a decrease in detection rate when ten peaches are swabbed with one foam disk. In order to maintain some consistency across the sample preparation process, a composite size of three units was selected. The varying topography of the commodities necessitated minor modifications to the method; for example, analysis of kiwi required that the hair on the surface be shaved prior to swabbing to achieve good recovery. Additionally, the effect of storage conditions on detection rate was analysed by spiking the surface of tomatoes at levels of 5 and 10 ng g(-1) for each pesticide, storing them under refrigeration and ambient conditions for 3 and 8 days, and then analysing the surface using this method. After 8 days of storage under both conditions more than 80% of the pesticides in the mixture were detected. Also, analysis of the multi-class mixtures was performed in both positive- and negative-ion mode and many classes were detected in both modes. Some classes, such thiocarbamates, phenylamides and organochlorine pesticides, were only observed in the positive-ion mode, whilst benzoylphenylurea pesticides were only detected in the negative-ion mode.


Cognitive and clinical predictors of work outcomes in clients with schizophrenia receiving supported employment services: 4-year follow-up.
In a prior study we showed that cognitive functioning was a modest predictor of work and supported employment services over 2-years in 30 clients with schizophrenia, whereas symptoms were not (McGurk et al. (2003). Psychiatric Services, 58, 1129-1135). In order to evaluate whether the long-term provision of supported employment services reduced the impact of cognitive functioning on work, we examined the relationships between cognitive functioning and symptoms assessed after the initial 2 years of the program, and work and vocational services over the following 2 years (3-4 years after joining the program). Cognitive functioning was more predictive of work during the latter 2 years of the study than the first 2 years, and a similar but weaker pattern was present for the prediction of employment services. Symptoms remained weak predictors for both time periods. In addition, learning and memory and executive functions were strongly correlated with job task complexity during the 3-4 year follow-up, but not the 1-2 year follow-up, suggesting that employment specialists were able to improve their ability to match clients to jobs based on their cognitive skills. Furthermore, the specific associations between cognitive functioning, services, and work outcomes changed from years 1-2 to years 3-4, suggesting a dynamic interplay between these factors over the long-term, rather than static and unchanging relationships. The findings indicate that rather than supported employment services reducing the impact of cognitive functioning on long-term competitive work, the impact actually increases over time, suggesting that efforts to improve cognitive functioning (e.g., cognitive rehabilitation) may optimize employment outcomes in schizophrenia.


Spoken word memory traces within the human auditory cortex revealed by repetition priming and functional magnetic resonance imaging.
Previous neuroimaging studies in the visual domain have shown that neurons along the perceptual processing pathway retain the physical properties of written words, faces, and objects. The aim of this study was to reveal the existence of similar neuronal properties within the human auditory cortex. Brain activity was measured using functional magnetic resonance imaging during a repetition priming paradigm, with words and pseudowords heard in an acoustically degraded format. Both the amplitude and peak latency of the hemodynamic response (HR) were assessed to determine the nature of the neuronal signature of spoken word priming. A statistically significant stimulus type by repetition interaction was found in various bilateral auditory cortical areas, demonstrating either HR suppression and enhancement for repeated spoken words and pseudowords, respectively, or word-specific repetition suppression without any significant effects for pseudowords. Repetition latency shift only occurred with word-specific repetition suppression in the right middle/posterior superior temporal sulcus. In this region, both repetition suppression and latency shift were related to behavioral priming. Our findings highlight for the first time the existence of long-term spoken word memory traces within the human auditory cortex. The timescale of auditory information integration and the neuronal mechanisms underlying priming both appear to differ according to the level of representations coded by neurons. Repetition may "sharpen" word-nonspecific representations coding short temporal variations, whereas a complex interaction between the activation strength and temporal integration of neuronal activity may occur in neuronal populations coding word-specific representations within longer temporal windows.


Smartphone and tablet apps for concussion road warriors (team clinicians): a systematic review for practical users.
Mobile technologies are steadily replacing traditional assessment approaches for the recognition and assessment of a sports concussion. Their ease of access, while facilitating the early identification of a concussion, also raises issues regarding the content of the applications (apps) and their suitability for different user groups. To locate and review apps that assist in the recognition and assessment of a sports concussion and to assess their content with respect to that of internationally accepted best-practice instruments. A search of international app stores and of the web using key terms such as 'concussion', 'sports concussion' and variants was conducted. For those apps meeting the inclusion criteria, data were extracted on the platform, intended users and price. The content of each app was benchmarked to the Sport Concussion Assessment Tool 2 (SCAT2) and Pocket SCAT2 using a custom scoring scheme to generate a percentage compliance statistic. 18 of the 155 apps identified met the inclusion criteria. Almost all (16/18) were available on an iOS platform and only five required a payment to purchase. The apps were marketed for a wide range of intended users from medical professionals to the general public. The content of the apps varied from 0% to 100% compliance with the selected standard, and 'symptom evaluation' components demonstrated the highest level of compliance. The surge in availability of apps in an unregulated market raises concerns as to the appropriateness of their content for different groups of end users. The consolidation of best-practice concussion instruments now provides a framework to inform the development of future apps.


Selective use of the electrocardiogram in pediatric preparticipation athletic examinations among pediatric primary care providers.
Recent literature examining insurance administrative data suggests that a selective approach, with concurrent history and physical exam (H&P), for obtaining an electrocardiogram (ECG) as a part of a preparticipation examination (PPE) for pediatric athletes is commonly used in the primary care setting demonstrating a high rate of disease detection. We sought to understand practice patterns of providers with regard to usage of ECG as a part of PPE. Utilizing an online survey, we queried primary care providers regarding their practice patterns, rationale, and concerns regarding use of ECGs as a part of the PPE. A total of 125 pediatricians completed the survey; 73.7% selectively use the ECG, 24.6% never use the ECG, and only 1.7% always obtain an ECG as part of the PPE. The most common rationale for selectively or never using the ECG is the belief that the H&P is sufficient to identify cardiac disease (70%). The most common H&P findings that lead to ECG screening include chest pain or syncope with exertion, family history of sudden cardiac death, an irregular heart rate, and a diastolic murmur. Among the diseases associated with sudden cardiac death, most pediatricians fear missing hypertrophic cardiomyopathy. Based on a survey of primary care providers, most practitioners are utilizing a selective approach of obtaining an ECG as a part of a PPE for athletic participation, which is in agreement with the current American Heart Association guidelines. Significant practice variation continues to exist, and may represent an area for future resource optimization.


Operation Pied Piper: a geographical reappraisal of the impact of wartime evacuation on scarlet fever and diphtheria rates in England and Wales, 1939-1945.
This paper examines the geographical impact of the British Government's wartime evacuation scheme on notified rates of two common acute childhood diseases (scarlet fever and diphtheria) in the 1470 local government districts of England and Wales, 1939-1945. Drawing on the notifications of communicable diseases collated by the General Register Office (GRO), we establish pre-war (baseline) disease rates for the 1470 districts. For the war years, techniques of binary logistic regression analysis are used to assess the associations between (a) above-baseline ('raised') disease rates in evacuation, neutral and reception districts and (b) the major phases of the evacuation scheme. The analysis demonstrates that the evacuation was temporally associated with distinct national and regional effects on notified levels of disease activity. These effects were most pronounced in the early years of the dispersal (1939-1941) and corresponded with initial levels of evacuation-related population change at the regional and district scales.


Effect of starch and sucrose on dental biofilm formation and on root dentine demineralization.
The cariogenicity of starch alone or in combination with sucrose is controversial and the effect on dentine demineralization and on the dental biofilm formed has not been explored under controlled conditions. A crossover, single-blind study was conducted in four steps of 14 days each, during which 11 volunteers wore palatal appliance containing 10 slabs of root dentine to which the following treatments were applied extraorally: 2% starch gel-like solution (starch group); 10% sucrose solution (sucrose group); a solution containing 2% starch and 10% sucrose (starch + sucrose group), or 2% starch solution followed by 10% sucrose solution (starch --> sucrose group). On the 14th day of each phase the biofilms were collected for biochemical and microbiological analyses, and dentine demineralization was assessed by hardness. A higher demineralization was found in dentine exposed to sucrose and starch sucrose combinations than to starch alone (p < 0.01), but the sucrose-containing groups did not differ significantly from each other (p > 0.05). The concentrations of soluble and insoluble extracellular polysaccharides (EPS), and the proportion of insoluble EPS, were lower in the biofilm formed in presence of starch (p < 0.01) than in those formed in the presence of sucrose or sucrose/starch combinations; however, no significant difference was observed among the groups containing sucrose (p > 0.05). RNA was successfully isolated and purified from in situ biofilms and only biofilms formed in response to sucrose and starch/sucrose combinations showed detectable levels of gtfB and gtfC mRNA. The findings suggest that the combination of starch with sucrose may not be more cariogenic to dentine than sucrose alone.


Perineural spread of malignant mesothelioma resulting in an intradural spinal cord mass: case report.
Spinal cord involvement by perineural spread of malignant mesothelioma is rare. We report a case of malignant mesothelioma that spread locally to invade the bony spine with both extradural and intradural perineural spread into the spinal canal that resulted in spinal cord compression. A 61-year-old man with a history of malignant mesothelioma presented with progressive leg weakness and right-sided arm weakness. Magnetic resonance imaging showed an enhancing lesion in the apex of the right lung with extension through the C7-T1 foramina with right hemicord enhancement. The patient underwent a C7-T1 laminectomy and right-sided C7-T1 and T1-T2 foraminotomies for neural decompression and biopsy of the lesion. Intraoperatively, tumor extended epidurally, and intradural perineural tumor spread along the C8 and T1 nerve roots into the spinal cord. Because it adhered to the spinal cord, no dissectible plane could be identified that would allow for safe total removal of the tumor. The epidural portion of the tumor, the adjacent involved bone, and the T1 nerve root were resected. Pathologic examination revealed malignant mesothelioma with bony invasion and perineural spread along the T1 nerve root. After decompression of the spinal cord, the patient had moderate improvement of his hand and leg function. Perineural spread of malignant mesothelioma resulting in spinal cord compression is an unusual clinical presentation. Intimate involvement of the spinal cord may prohibit aggressive tumor resection.


pagP is required for resistance to antibody-mediated complement lysis during Bordetella bronchiseptica respiratory infection.
To efficiently colonize and persist in the lower respiratory tract, bacteria must survive multiple host immune mechanisms. Bordetella bronchiseptica is a gram-negative respiratory pathogen that naturally infects mice and persists in the lower respiratory tract for up to 49 days postinoculation. In this work, we examined the effect of mutation of the pagP gene on the persistence of B. bronchiseptica in the lower respiratory tract of mice. The pagP gene encodes a palmitoyl transferase that is responsible for the addition of a palmitoyl group to the lipid A region of B. bronchiseptica lipopolysaccharide. Data presented here confirm that a B. bronchiseptica deltapagP mutant demonstrates defective persistence in the lower respiratory tract of wild-type mice. We hypothesized that the defective persistence of the B. bronchiseptica deltapagP mutant was due to an increased susceptibility of this mutant to a host immune response. In vivo data indicate that both B cells and the complement component C3 are required for the reduced bacterial numbers of the deltapagP mutant on day 14 postinoculation. In addition, an in vitro complement killing assay demonstrated that B. bronchiseptica exhibits pagP-dependent resistance to antibody-mediated complement killing at low concentrations of immune serum. Taken together, these results suggest that pagP is required for B. bronchiseptica to resist antibody-mediated complement lysis during respiratory infection.


"It's all about trust": reflections of researchers on the complexity and controversy surrounding biobanking in South Africa.
Biobanks are precariously situated at the intersection of science, genetics, genomics, society, ethics, the law and politics. This multi-disciplinarity has given rise to a new discourse in health research involving diverse stakeholders. Each stakeholder is embedded in a unique context and articulates his/her biobanking activities differently. To researchers, biobanks carry enormous transformative potential in terms of advancing scientific discovery and knowledge. However, in the context of power asymmetries in Africa and a distrust in science born out of historical exploitation, researchers must balance the scientific imperative of collecting, storing and sharing high quality biological samples with obligations to donors/participants, communities, international collaborators, regulatory and ethics authorities. To date, researcher perspectives on biobanking in South Africa have not been explored and documented. In-depth qualitative interviews were conducted with a purposive sample of 21 researchers - 8 in the Western Cape, 3 in Gauteng and 10 in Kwa-Zulu Natal. Interviews lasted approximately 40-60 min and were audiotaped with consent. Thematic analysis of the transcribed interviews was conducted by the co-authors. Researchers articulated serious concerns over standardised regulatory approaches that failed to consider the heterogeneity of biobanks. Given that biobanks differ considerably, guidelines and RECs need to stratify risk accordingly and governance processes and structures must be flexible. While RECs were regarded as an important component of the governance structure researchers expressed concern about their expertise in biobanking. Operational management of biobanks was regarded as an ethical imperative and a pre-requisite to building trust during consent processes. While broad general consent was preferred, tiered consent was thought to be more consistent with respect for autonomy and building trust. Material Transfer Agreements (MTAs) were often lacking when biosamples were exported and this was perceived to impact negatively on trust. On the other hand, researchers believed that authentic community engagement would help to build trust. Building trust will best be achieved via a system of governance structures and processes that precede the establishment of a biobank and monitor progress from the point of sample collection through to future use, including export. Such governance structures must be robust and must include comprehensive national legislation, policy and contextualised guidelines. Currently such governance infrastructure appears to be lacking in many African countries including South Africa. Capacity development of all stakeholders including REC members will enhance expeditious and efficient review of biobanking protocols which in turn will reinforce trust in the researcher-donor relationship. Science translation and community engagement in biobanking is integral to the success of biobanking in South Africa.


Loss-of-function analysis reveals distinct requirements of the translation initiation factors eIF4E, eIF4E-3, eIF4G and eIF4G2 in Drosophila spermatogenesis.
In eukaryotes, post-transcriptional regulation of gene expression has a key role in many cellular and developmental processes. Spermatogenesis involves a complex developmental program that includes changes in cell cycle dynamics and dramatic cellular remodeling. Translational control is critical for spermatogenesis in Drosophila as many mRNAs synthesized in the spermatocytes are translated only much later during spermatid differentiation. Testes-specific translation initiation factors eIF4E-3 and eIF4G2 are essential specifically for male fertility. However, details of their roles during different stages of spermatogenesis are unknown, and the role of canonical translation initiation factors in spermatogenesis remains unexplored. In this study, we addressed the functional role of eIF4E-1, eIF4E-3, eIF4G and eIF4G2 in testes development and formation of mature sperm. Using the UAS-Gal4 system and RNA interference, we systematically knocked down these four genes in different stages of germ cell development, and in the somatic cells. Our results show that eIF4E-1 function in early germ cells and the surrounding somatic cells is critical for spermatogenesis. Both eIF4E-1 and eIF4E-3 are required in spermatocytes for chromosome condensation and cytokinesis during the meiotic stages. Interestingly, we find that eIF4G knockdown did not affect male fertility while eIF4G2 has distinct functions during spermatogenesis; it is required in early germ cells for proper meiotic divisions and spermatid elongation while its abrogation in spermatocytes caused meiotic arrest. Double knockdown of eIF4G and eIF4G2 shows that these proteins act redundantly during the early stages of spermatogenesis. Taken together, our analysis reveals spatio-temporal roles of the canonical and testes-specific translation initiation factors in coordinating developmental programs during spermatogenesis.