Transition Metal Complexes of Calix[4]arene: Theoretical Investigations into Small Guest Binding within the Host Cavity.
The ability to selectively detect or store small molecules, such as gases, is of enormous commercial potential. Calixarenes have been studied extensively as host molecules; however, recent synthetic advances have seen the formation of new polymetallic calixarene clusters, which have not yet been explored for such purposes. We therefore present a theoretical study, using Density Functional Theory, to thoroughly investigate the binding preferences of calix[4]arene, with a variety of transition metal cations coordinated to the calixarene tetraphenolic pocket, toward a series of important small molecules, H2S, SO2, H2O, O2, H2, N2, N2O, CO2, NH3, and HCN. It was found that the inclusion of a metal atom at the lower-rim of the calixarene caused significant strengthening of binding energy with all of the small molecules in our study as compared to metal-free calixarene. The guests, SO2 and NH3, were found to bind strongest with H2 binding weakest. Our calculations predict that simply introducing metal coordination of any type to calix[4]arene will make the largest difference to the binding energies. Subsequently changing the type, oxidation state, or the spin state of the metal coordinated to the calixarene tetraphenolic pocket was found to have a lesser effect on these.


FERM domain mutations induce gain of function in JAK3 in adult T-cell leukemia/lymphoma.
Adult T-cell leukemia/lymphoma (ATLL) is an incurable disease where most patients succumb within the first year of diagnosis. Both standard chemotherapy regimens and mAbs directed against ATLL tumor markers do not alter this aggressive clinical course. Therapeutic development would be facilitated by the discovery of genes and pathways that drive or initiate ATLL, but so far amenable drug targets have not been forthcoming. Because the IL-2 signaling pathway plays a prominent role in ATLL pathogenesis, mutational analysis of pathway components should yield interesting results. In this study, we focused on JAK3, the nonreceptor tyrosine kinase that signals from the IL-2R, where activating mutations have been found in diverse neoplasms. We screened 36 ATLL patients and 24 ethnically matched controls and found 4 patients with mutations in JAK3. These somatic, missense mutations occurred in the N-terminal FERM (founding members: band 4.1, ezrin, radixin, and moesin) domain and induced gain of function in JAK3. Importantly, we show that these mutant JAK3s are inhibited with a specific kinase inhibitor already in human clinical testing. Our findings underscore the importance of this pathway in ATLL development and offer a therapeutic handle for this incurable cancer.


Application of novel hemostatic agent during laparoscopic partial nephrectomy.
We determined if QuikClot, a novel hemostatic agent made of a granulated mineral substance, could be used to control renal parenchymal bleeding and collecting system leakage during open and laparoscopic partial nephrectomy. After obtaining renal hilar vascular control 2 domestic female pigs underwent bilateral open and 4 underwent unilateral laparoscopic partial nephrectomy. After excision of the lower pole without cautery the hemostatic agent was applied to the cut surface of the kidney and hilar vascular control was released. Additional QuikClot was added until complete hemostasis was achieved. One week postoperatively the animals were sacrificed and the operated kidneys were harvested for ex vivo retrograde pyelograms and histopathological analysis. All partial nephrectomies were performed without complication. Mean operative and warm ischemia times were 62 and 16 minutes, respectively. An average of 23% of renal mass by weight was resected with a mean blood loss of 73 ml per procedure. No cautery, additional hemostatic agents or techniques were used. No animal had clinical or radiographic evidence of urinoma or delayed hemorrhage. Histopathological analysis showed preservation of the renal parenchyma immediately beneath the QuikClot layer. In the porcine model QuikClot allowed the resection of large renal segments, while providing reliable hemostasis and closure of the renal collecting system. No deleterious effect on underlying renal parenchyma or surrounding tissues was observed.


Eugenics, medical education, and the Public Health Service: Another perspective on the Tuskegee syphilis experiment.
The Public Health Service (PHS) Study of Untreated Syphilis in the Male Negro (1932-72) is the most infamous American example of medical research abuse. Commentary on the study has often focused on the reasons for its initiation and for its long duration. Racism, bureaucratic inertia, and the personal motivations of study personnel have been suggested as possible explanations. We develop another explanation by examining the educational and professional linkages shared by three key physicians who launched and directed the study. PHS surgeon general Hugh Cumming initiated Tuskegee, and assistant surgeons general Taliaferro Clark and Raymond A. Vonderlehr presided over the study during its first decade. All three had graduated from the medical school at the University of Virginia, a center of eugenics teaching, where students were trained to think about race as a key factor in both the etiology and the natural history of syphilis. Along with other senior officers in the PHS, they were publicly aligned with the eugenics movement. Tuskegee provided a vehicle for testing a eugenic hypothesis: that racial groups were differentially susceptible to infectious diseases.


Antibiotic Elution and Mechanical Strength of PMMA Bone Cement Loaded With Borate Bioactive Glass.
Introduction: Local delivery of antibiotics using bone cement as the delivery vehicle is an established method of managing implant-associated orthopedic infections. Various fillers have been added to cement to increase antibiotic elution, but they often do so at the expense of strength. This study evaluated the effect of adding a borate bioactive glass, previously shown to promote bone formation, on vancomycin elution from PMMA bone cement. Methods: Five cement composites were made: three loaded with borate bioactive glass along with 0, 1, and 5 grams of vancomycin and two without any glass but with 1 and 5 grams vancomycin to serve as controls. The specimens were soaked in PBS. Eluate of vancomycin was collected every 24 hours and analyzed by HPLC. Orthopedic-relevant mechanical properties of each composite were tested over time. Results: The addition of borate bioactive glass provided an increase in vancomycin release at Day 1 and an increase in sustained vancomycin release throughout the treatment period. An 87.6% and 21.1% increase in cumulative vancomycin release was seen for both 1g and 5g loading groups, respectively. Compressive strength of all composites remained above the weight-bearing threshold of 70 MPa throughout the duration of the study with the glass-containing composites showing comparable strength to their respective controls. Conclusion: The incorporation of borate bioactive glass into commercial PMMA bone cement can significantly increase the elution of vancomycin. The mechanical strength of the cement-glass composites remained above 70 MPa even after soaking for 8 weeks, suggesting their suitability for orthopedic weight-bearing applications.


Synaptic competition in structural plasticity and cognitive function.
Connections between neurons can undergo long-lasting changes in synaptic strength correlating with changes in structure. These events require the synthesis of new proteins, the availability of which can lead to cooperative and competitive interactions between synapses for the expression of plasticity. These processes can occur over limited spatial distances and temporal periods, defining dendritic regions over which activity may be integrated and could lead to the physical rewiring of synapses into functional groups. Such clustering of inputs may increase the computational power of neurons by allowing information to be combined in a greater than additive manner. The availability of new proteins may be a key modulatory step towards activity-dependent, long-term growth or elimination of spines necessary for remodelling of connections. Thus, the aberrant growth or shrinkage of dendritic spines could occur if protein levels are misregulated. Indeed, such perturbations can be seen in several mental retardation disorders, wherein either too much or too little protein translation exists, matching an observed increase or decrease in spine density, respectively. Cellular events which alter protein availability could relieve a constraint on synaptic competition and disturb synaptic clustering mechanisms. These changes may be detrimental to modifications in neural circuitry following activity.


A NIMA-related protein kinase is essential for completion of the sexual cycle of malaria parasites.
The molecular mechanisms regulating the sexual development of malaria parasites from gametocytes to oocysts in their mosquito vector are still largely unexplored. In other eukaryotes, NIMA-related kinases (Neks) regulate cell cycle progression and have been implicated in the regulation of meiosis. Here, we demonstrate that Nek-4, a new Plasmodium member of the Nek family, is essential for completion of the sexual cycle of the parasite. Recombinant Plasmodium falciparum Nek-4 possesses protein kinase activity and displays substrate preferences similar to those of other Neks. Nek-4 is highly expressed in gametocytes, yet disruption of the nek-4 gene in the rodent malaria parasite P. berghei has no effect on gamete formation and subsequent fertilization. However, further differentiation of zygotes into ookinetes is abolished. Measurements of nuclear DNA content indicate that zygotes lacking Nek-4 fail to undergo the genome replication to the tetraploid level that precedes meiosis. Cell cycle progression in the zygote is identified as a likely precondition for its morphological transition to the ookinete and for the successful establishment of a malaria infection in the mosquito.


Preparative Scale Production of Recombinant Human Transthyretin for Biophysical Studies of Protein-Ligand and Protein-Protein Interactions.
Human transthyretin (hTTR), a serum protein with a main role in transporting thyroid hormones and retinol through binding to the retinol-binding protein, is an amyloidogenic protein involved in familial amyloidotic polyneuropathy (FAP), familial amyloidotic cardiomyopathy, and central nervous system selective amyloidosis. hTTR also has a neuroprotective role in Alzheimer disease, being the major Aβ binding protein in human cerebrospinal fluid (CSF) that prevents amyloid-β (Aβ) aggregation with consequent abrogation of toxicity. Here we report an optimized preparative expression and purification protocol of hTTR (wt and amyloidogenic mutants) for in vitro screening assays of TTR ligands acting as amyloidogenesis inhibitors or acting as molecular chaperones to enhance the TTR:Aβ interaction. Preparative yields were up to 660 mg of homogenous protein per L of culture in fed-batch bioreactor. The recombinant wt protein is mainly unmodified at Cys10, the single cysteine in the protein sequence, whereas the highly amyloidogenic Y78F variant renders mainly the S-glutathionated form, which has essentially the same amyloidogenic behavior than the reduced protein with free Cys10. The TTR production protocol has shown inter-batch reproducibility of expression and protein quality for in vitro screening assays.


Machines, medication, modulation: circuits of dependency and self-care in Las Vegas.
The intensive entertainment infrastructure of Las Vegas is overlaid with a robust therapeutic network for those who become addicted to its technologies. Although the objectives of gambling machines and addiction therapeutics are seemingly at odds--the first work to encourage play, the second to stop it--both gear their interventions around a model of the self as a continuum of behavioral potentials that can be externally modulated. For compulsive gamblers implicated in this circuit of modulation, pharmaceutical drugs that have been prescribed to dampen cravings for machine play sometimes function as intensifiers of its effects. Caught in an intractable play between technologies of harm and technologies of care, recovering gambling addicts are challenged to assemble a technical array through which they can maintain balance; health itself, for these individuals, becomes a state of managed dependency. This essay explores the shifting terms and changing stakes of subjectivity and health in the contemporary United States by way of ethnographic research on compulsive gamblers who live and work in Las Vegas. The analysis draws on interviews with gamblers as well as on observations in local self-help groups, directed group therapy sessions, and chat rooms of Internet recovery Web sites.


Growth and hepatic in vitro metabolism of ergotamine in mice divergently selected for response to endophyte toxicity.
This study investigated if genetic differences exhibited in endophyte-resistant and -susceptible mouse lines had persisted after 13 generations in which the integrity of lines was maintained yet selection ceased. Experimental groups were mouse lines fed an endophyte-free (E-) or -infected (E+) diet. The in vitro metabolism of the ergot alkaloid ergotamine in mouse liver microsomes was characterized by LC-MS/MS and compared between both lines before and after exposure to E+ feed. No difference in the average daily weight gain of pups between resistant and susceptible mice was observed on the E+ diet. Thus, for the weight gain selection criterion under study, the difference established between the two lines appears not to have persisted over the extended period of relaxed selection. Microsomal incubations produced nine predominate peaks in the HPLC assay. The peaks were confirmed by LC-MS/MS to be ergotamine, ergotamine epimer, monohydroxylated metabolites (M1, M2, M1e, M2e) and dihydroxylated metabolites (M3--5). A gender difference for metabolite formation was observed on the E- diet, in that females produced a greater amount of M1, M1e and M3--5 than males. When challenged with the E+ diet, mice showed differences in concentration of M3 for line (resistant > susceptible) and gender (female > male) and of M4 and M5 for gender (female > male). Gender differences in the metabolism of ergotamine have not been shown before in these lines of mice or other species used to study ergot alkaloid metabolism. This adds a potential source of variation in the susceptibility to fescue toxicity not explored previously and would be of value to investigate further.


Development of a stable Leishmania expression vector and application to the study of parasite surface antigen genes.
Trypanosomatid protozoan parasites cause several important tropical diseases and have been a fertile ground for the discovery of molecular paradigms such as trans-splicing and RNA editing. Transfection-based methods for the study of these organisms have recently been developed, and we have now designed an expression vector, pX, which contains only 2.3 kilobases of Leishmania DNA and can be stably transfected with high efficiency. Genes encoding Escherichia coli beta-galactosidase or a Leishmania amazonensis protective membrane glycoprotein (GP46A/M-2) were inserted into the pX expression site and transfected into Leishmania major, where they directed the synthesis of high levels of mRNAs formed by 5' and 3' processing events occurring predominantly at the sites used by the normal transcripts. Colony assays and immunoblot analysis showed that both proteins were produced; enzymatically active beta-galactosidase comprised approximately 1% of total protein. Sizes of the GP46A protein synthesized in transfected L. major or L. amazonensis were similar and differed from the predominant L. amazonensis GP46, suggesting that the GP46A gene may encode a variant GP46 family member. Because these vectors function efficiently in pathogenic species of Leishmania, pX will facilitate the genetic analyses of parasite proteins crucial for infectivity as well as the identification of cis-acting elements mediating transcription and replication.


Genetic factors in familial manifestation of primary mediastinal large B-cell lymphoma over two generations.
Primary mediastinal large B-cell lymphoma (PMBL) is a rarely occurring lymphoid malignancy, which typically affects young adults and presents itself as an anterior mediastinal mass. Gene expression profiling, as well as somatic genetic analysis revealed that it is closely related to classical Hodgkin lymphoma, whereas morphologically it tends to resemble diffuse large B-cell lymphoma. Familial clustering of PMBL is rare - only two reports have been published to date. While generally accepted that positive family history is associated with increased risk of developing a lymphoma, genetic risk factors, which might predispose to PMBL are largely unknown. We performed germline and tumor genetic analysis by whole exome sequencing and array-CGH of a family, in which the father and the son both developed a PMBL. Germline investigations of both affected patients and of their two unaffected family members has not been able to provide a single risk factor associated with lymphoma predisposition. In addition, genes that were previously implicated in increased risk for PMBL, namely MLL (KMT2A) and TIRAP, were found to be intact in all investigated family members. Somatic genetic investigations identified known as well as novel genetic aberrations in tumors of affected subjects. We conclude that predisposition to a PMBL might be inherited through a combination of low- or moderate-risk factors and provide a shortlist of most likely selected candidates, which can be used in future studies.


Spin-dependent operators in correlated Gaussian bases.
In their textbook, Suzuki and Varga [(Springer, Berlin, 1998)] present the stochastic variational method with the correlated Gaussian basis in a very exhaustive way. The matrix elements for central potentials are expressed in an elegant form, but the elements for spin-dependent operators, when treated, are given as very cumbersome expressions. We find many formulas for those elements. Their expressions are given in terms of the same geometrical functions that appear in the case of central potentials. These functions obtain therefore a universal status; this property is very useful for numerical applications.


RNAi and gene therapy: a mutual attraction.
The phylogenetically conserved cellular phenomenon of RNA interference (RNAi)-the sequence-specific post-transcriptional silencing of gene expression mediated by small double-stranded RNAs-holds substantial promise for basic research and for drug development. Particularly attractive from a medical standpoint is the juxtaposition of new RNAi methodology with established gene transfer strategies, especially viral vectors for efficient and tissue-specific RNAi delivery to patients. Here, we summarize the latest experimental and clinical advances in RNAi-based gene therapy approaches. We briefly portray emerging nonviral strategies for siRNA transfer, before comparing the three viral vectors currently predominantly developed as shRNA delivery vehicles, adenovirus, lentivirus, and adeno-associated virus (AAV). Moreover, we describe the most clinically relevant genetic, acquired or infectious targets being pursued for therapeutic purposes. Specifically, we assess the use of vector-mediated RNAi for treatment of viral processes, solid cancers, lymphoproliferative disorders, and neurodegenerative and ocular diseases. In addition, we highlight further emerging applications, including stem cell therapies and animal transgenesis, as well as discuss some of the potential pitfalls and limitations inherent to the individual approaches. While we predict that eventual schemes will be shaped by our increasing understanding of the complexities of human RNAi biology, as well as by progressive refinements of viral shuttle designs, the potential scientific and medical benefits from a successful marriage of RNAi and gene therapy seem enormous.


The Mus81/Mms4 endonuclease acts independently of double-Holliday junction resolution to promote a distinct subset of crossovers during meiosis in budding yeast.
Current models for meiotic recombination require that crossovers derive from the resolution of a double-Holliday junction (dHJ) intermediate. In prokaryotes, enzymes responsible for HJ resolution are well characterized but the identification of a eukaryotic nuclear HJ resolvase has been elusive. Indirect evidence suggests that MUS81 from humans and fission yeast encodes a HJ resolvase. We provide three lines of evidence that Mus81/Mms4 is not the major meiotic HJ resolvase in S. cerevisiae: (1) MUS81/MMS4 is required to form only a distinct subset of crossovers; (2) rather than accumulating, dHJ intermediates are reduced in an mms4 mutant; and (3) expression of a bacterial HJ resolvase has no suppressive effect on mus81 meiotic phenotypes. Our analysis also reveals the existence of two distinct classes of crossovers in budding yeast. Class I is dependent upon MSH4/MSH5 and exhibits crossover interference, while class II is dependent upon MUS81/MMS4 and exhibits no interference. mms4 specifically reduces crossing over on small chromosomes, which are known to undergo less interference. The correlation between recombination rate and degree of interference to chromosome size may therefore be achieved by modulating the balance between class I/class II crossovers.


Detection of all four dengue serotypes in Aedes aegypti female mosquitoes collected in a rural area in Colombia.
The Aedes aegypti vector for dengue virus (DENV) has been reported in urban and periurban areas. The information about DENV circulation in mosquitoes in Colombian rural areas is limited, so we aimed to evaluate the presence of DENV in Ae. aegypti females caught in rural locations of two Colombian municipalities, Anapoima and La Mesa. Mosquitoes from 497 rural households in 44 different rural settlements were collected. Pools of about 20 Ae. aegypti females were processed for DENV serotype detection. DENV in mosquitoes was detected in 74% of the analysed settlements with a pool positivity rate of 62%. The estimated individual mosquito infection rate was 4.12% and the minimum infection rate was 33.3/1,000 mosquitoes. All four serotypes were detected; the most frequent being DENV-2 (50%) and DENV-1 (35%). Two-three serotypes were detected simultaneously in separate pools. This is the first report on the co-occurrence of natural DENV infection of mosquitoes in Colombian rural areas. The findings are important for understanding dengue transmission and planning control strategies. A potential latent virus reservoir in rural areas could spill over to urban areas during population movements. Detecting DENV in wild-caught adult mosquitoes should be included in the development of dengue epidemic forecasting models.


"If He Has it, We Know What to Do": Parent Perspectives on Familial Risk for Autism Spectrum Disorder.
Predictive testing for familial disorders can guide healthcare and reproductive decisions. Familial disorders with onset in childhood (e.g., autism spectrum disorder [ASD]) are promising targets for presymptomatic prediction; however, little is known about parent perceptions of risk to their children in the presymptomatic period. The current study examined risk perceptions in parents of infants at high familial risk for ASD enrolled in a longitudinal study of brain and behavior development. Semistructured interviews were conducted with 37 parents of high-risk infants during the presymptomatic window (3-15 months) that precedes an ASD diagnosis. Infants were identified as high familial risk due to having an older sibling with ASD. Parent interview responses were coded and interpreted to distill emerging themes. The majority of parents were aware of the increased risk of ASD for their infants, and risk perceptions were influenced by comparisons to their older child with ASD. Parents reported a variety of negative emotions in response to perceived risk, including worry, fear, and sadness, and described impacts of perceived risk on their behavior: increased vigilance to emerging symptoms, altered reproductive and healthcare decisions, and seeking ongoing assessment through research. Parents of children at high familial risk for childhood-onset disorders like ASD face a period of challenging uncertainty during early development. In anticipation of a future in which presymptomatic testing for ASD is made available, it is important to understand how parents react to and cope with the elevated-but still highly uncertain-risk conveyed by family history.


Small eyes (Sey): a homozygous lethal mutation on chromosome 2 which affects the differentiation of both lens and nasal placodes in the mouse.
Small eyes (Sey) is a semidominant, homozygous lethal mutation in the mouse (Roberts, 1967). It is allelic with SeyH, a radiation-induced homozygous prenatal lethal which has been mapped on chromosome 2. The effect of the Sey mutation is apparently limited to the growth and differentiation of the presumptive lens and nasal placodes. Homozygous Sey/Sey embryos can be distinguished as early as 10.5 days post coitum (p.c.); the optic vesicles grow out, but the ectoderm does not give rise to a lens and nasal pits never form. Immunohistochemical studies show that the distribution of the extracellular matrix glycoprotein laminin is not significantly different in the cephalic region of Sey/Sey versus Sey/+ or +/+ embryos. Sey/Sey embryos develop to term but without eyes or nose, and die soon after birth. Further analysis of Sey/Sey embryos may throw light on the mechanisms underlying morphogenesis of craniofacial structures in mammals.


Laparoscopy in the management of patients with ovarian carcinoma.
Since operative evaluations of patients undergoing therapy for ovarian cancer are necessary, the laparoscope will continue to play a role in the management of these patients. However, the sensitivity of the laparoscope for detecting disease either prior to or at the completion of chemotherapy is significantly less than that of laparotomy, and therefore its utility must be more narrowly defined. The morbidity of laparoscopy is low when performed correctly and utilizing techniques to avoid bowel perforation. Most patients require only a brief hospitalization, and those who cannot medically tolerate an extensive laparotomy might be able to undergo a laparoscopy. The major limitations of laparoscopy are the inability to adequately inspect the peritoneal cavity and pelvis in at least one-quarter of patients because of extensive adhesions, to assess retroperitoneal lymph nodes in all patients, and to resect tumour masses necessary to cytoreduce the tumour prior to therapy. The successful performance of interval laparoscopy, however, is predictive of survival and can select a group of patients whose likelihood of recurrence is relatively low (30 per cent) after more than three years. The observation that the majority of patients who have a negative laparoscopy following six months of chemotherapy remain free of disease for several years, suggests that sensitive tumours are most likely to respond during the initial courses of chemotherapy, and that consideration should be given to briefer and more intensive courses of therapy prior to operative evaluation of the patients.


Management of children visiting the emergency department during out-of-office hours: an observational study.
The aim was to study the characteristics and management of children visiting the emergency department (ED) during out-of-office hours. We analysed electronic health record data from 119 204 children visiting one of five EDs in four European countries. Patient characteristics and management (diagnostic tests, treatment, hospital admission and paediatric intensive care unit admission) were compared between children visiting during office hours and evening shifts, night shifts and weekend day shifts. Analyses were corrected for age, gender, Manchester Triage System urgency, abnormal vital signs, presenting problems and hospital. Patients presenting at night were younger (median (IQR) age: 3.7 (1.4-8.2) years vs 4.8 (1.8-9.9)), more often classified as high urgent (16.3% vs 9.9%) and more often had ≥2 abnormal vital signs (22.8% vs 18.1%) compared with office hours. After correcting for disease severity, laboratory and radiological tests were less likely to be requested (adjusted OR (aOR): 0.82, 95% CI 0.78-0.86 and aOR: 0.64, 95% CI 0.60-0.67, respectively); treatment was more likely to be undertaken (aOR: 1.56, 95% CI 1.49-1.63) and patients were more likely to be admitted to the hospital (aOR: 1.32, 95% CI 1.24-1.41) at night. Patterns in management during out-of-office hours were comparable between the different hospitals, with variability remaining. Children visiting during the night are relatively more seriously ill, highlighting the need to keep improving emergency care on a 24-hour-a-day basis. Further research is needed to explain the differences in management during the night and how these differences affect patient outcomes.


Tubular reabsorption rates for myoglobin in the isolated perfused rat kidney.
The renal handling of myoglobin has been studied in the isolated perfused rat kidney. Myoglobin was freely filtered. Reabsorption by the renal tubules showed saturation kinetics with a relatively low maximum rate of reabsorption (Tmax) of 27-30 micrograms min-1 g-1 wet wt. at a perfusate concentration of 70-80 micrograms/ml. Myoglobin reabsorption is therefore much less than that reported for immunoglobulin light chain or lysozyme in this model. Large increases in sodium and water excretion produced by omission of oncotic agent from the perfusate did not alter the kinetics of myoglobin reabsorption. The use of bovine serum albumin as oncotic agent in the perfusate prevented the tubular reabsorption of myoglobin. Small amounts of albumin are filtered by the isolated perfused kidney and it is postulated that this albumin interferes with tubular reabsorption of myoglobin.


Developmental febrile seizures modulate hippocampal gene expression of hyperpolarization-activated channels in an isoform- and cell-specific manner.
Febrile seizures, in addition to being the most common seizure type of the developing human, may contribute to the generation of subsequent limbic epilepsy. Our previous work has demonstrated that prolonged experimental febrile seizures in the immature rat model increased hippocampal excitability long term, enhancing susceptibility to future seizures. The mechanisms for these profound proepileptogenic changes did not require cell death and were associated with long-term slowed kinetics of the hyperpolarization-activated depolarizing current (I(H)). Here we show that these seizures modulate the expression of genes encoding this current, the hyperpolarization-activated, cyclic nucleotide-gated channels (HCNs): In CA1 neurons expressing multiple HCN isoforms, the seizures induced a coordinated reduction of HCN1 mRNA and enhancement of HCN2 expression, thus altering the neuronal HCN phenotype. The seizure-induced augmentation of HCN2 expression involved CA3 in addition to CA1, whereas for HCN4, mRNA expression was not changed by the seizures in either hippocampal region. This isoform- and region-specific transcriptional regulation of the HCNs required neuronal activity rather than hyperthermia alone, correlated with seizure duration, and favored the formation of slow-kinetics HCN2-encoded channels. In summary, these data demonstrate a novel, activity-dependent transcriptional regulation of HCN molecules by developmental seizures. These changes result in long-lasting alteration of the HCN phenotype of specific hippocampal neuronal populations, with profound consequences on the excitability of the hippocampal network.


Demographic Variations in Tinnitus Subjects with and without Hearing Loss: A Study of 175 Subjects.
This research was conducted to investigate the presenting features of tinnitus in subjects with normal hearing thresholds as compared to the ones with hearing loss. Sample population comprised of 175 subjects with tinnitus, in the age range of 18 to 55 years, segregated into two groups: G1 (75 subjects) having normal hearing with tinnitus and G2 (100 subjects) having hearing loss with tinnitus. All the subjects underwent conventional audiometric testing along with a thorough evaluation of all the parameters of tinnitus. Significantly large number of males reported with tinnitus as compared to females. Tinnitus was most prevailing in left ear and these subjects sought intervention earlier than right tinnitus subjects. G1 subjects reported earlier as compared to G2 subjects. Males looked out for specialist's advice earlier than females in both the groups. Large number of subjects in both groups reported with sudden onset and continuous tinnitus.


When two is too many: Collaborative encoding impairs memory.
Humans routinely encode and retrieve experiences in interactive, collaborative contexts. Yet much of what we know about human memory comes from research on individuals working in isolation. Some recent research has examined collaboration during retrieval, but not much is known about how collaboration during encoding affects memory. We examined this issue. Participants created episodes by elaborating on study materials alone or collaboratively, and they later performed a cued-recall task alone, with the study partner, or with a different partner (Experiment 1). Collaborative encoding impaired recall. This counterintuitive outcome was found for both individual and group recall, even when the same partners collaborated across encoding and retrieval. This impairment was significantly reduced, but persisted, when the encoding instructions encouraged free-flowing collaboration (Experiment 2). Thus, the collaborative-encoding deficit is robust in nature and likely occurs because collaborative encoding produces less effective cues for later retrieval.


Large-scale, polymerase chain reaction-based surveillance of Schistosoma haematobium DNA in snails from transmission sites in coastal Kenya: a new tool for studying the dynamics of snail infection.
Levels of prepatent Schistosoma haematobium infection were monitored in intermediate host snails (Bulinus nasutus) collected from transmission sites in coastal Kenya, using a polymerase chain reaction (PCR) assay amplifying the Dra I repeated sequence of S. haematobium. The timing and number of prepatent and patent infections were determined for each site and, where the time of first appearance was clear, the minimal prepatent period was estimated to be five weeks. High, persistent, prepatency rates (range = 28-54%), indicated a significant degree of repeated area contamination with parasite ova. In contrast, rates of cercarial shedding proved locally variable, and were either low (range = 0.14-3.4%) or altogether absent, indicating that only a small proportion of infected snails reach the stage of cercarial shedding. Given the apparently strong focal effects of environmental conditions, implications of these new data are discussed regarding the estimation of local force of transmission and the design of control activities.


A patient with scirrhous stomach cancer treated with combination of hyperthermotherapy and 5-aminolevulinic acid (ALA).
A 35-year-old female with scirrhous stomach cancer (stage IV) was treated with a combination of 5-aminolevulinic acid (ALA), sodium dichloroacetate (DCA), hyperthermotherapy, and immunotherapy as terminal care. The patient survived for one year and seven months, during which her quality of life was markedly improved and she returned to work. The patient was diagnosed with poorly-differentiated adenocarcinoma and progressive signet-ring cell carcinoma, accompanied by left ovarian metastasis, peritoneal dissemination, and right hydronephrosis stage IV, and treated with combination chemotherapy with tegafur-gimeracil-oteracil potassium (TS-1) and docetaxel. Oral ALA and DCA were concomitantly administered at 50 mg each three times a day (150 mg/day, respectively). In addition, hyperthermotherapy using thermotron was concomitantly performed at 2- to 3-week intervals. Cellular immunotherapy with αβ T- and immature dendritic cells was also performed. The disease did not progress for 11 months, her quality of life was markedly improved, and she was able to return to work. However, the signs of enlargement of the ovarian metastatic lesion were noted later, for which chemotherapy with four cycles of second-line paclitaxel and a half dose of irinotecan and cisplatin as third-line treatment were performed. Combination of ALA/DCA, hyperthermotherapy, and cellular immunotherapy may be a low-invasive palliative therapy superior in maintaining quality of life of tumor-bearing terminally ill individuals.


Correcting India's Chronic Shortage of Drug Inspectors to Ensure the Production and Distribution of Safe, High-Quality Medicines.
Good drug regulation requires an effective system for monitoring and inspection of manufacturing and sales units. In India, despite widespread agreement on this principle, ongoing shortages of drug inspectors have been identified by national committees since 1975. The growth of India's pharmaceutical industry and its large export market makes the problem more acute. The focus of this study is a case study of Maharashtra, which has 29% of India's manufacturing units and 38% of its medicines exports. India's regulations were reviewed, comparing international, national and state inspection norms with the actual number of inspectors and inspections. Twenty-six key informant interviews were conducted to ascertain the causes of the shortfall. In 2009-2010, 55% of the sanctioned posts of drug inspectors in Maharashtra were vacant. This resulted in a shortfall of 83%, based on the Mashelkar Committee's recommendations. Less than a quarter of the required inspections of manufacturing and sales units were undertaken. The Indian Drugs and Cosmetics Act and its Rules and Regulations make no provisions for drug inspectors and workforce planning norms, despite the growth and increasing complexity of India's pharmaceutical industry. The Maharashtra Food and Drug Administration (FDA) falls short of the Mashelkar Committee's recommended workforce planning norms. Legislation and political and operational support are required to produce needed changes.


Influence of pirenzepine on gallbladder contraction in man induced by sham feeding or an intraduodenal meal.
Pirenzepine, an M1 muscarinic receptor antagonist, was tested for its ability to antagonize sham feeding- and intraduodenal fatty meal-stimulated gallbladder contraction in man. Intravenously administered pirenzepine abolished sham feeding-induced gallbladder contraction. Pirenzepine also inhibited contraction induced by intraduodenal meal, but this inhibition was of a lesser magnitude than the inhibition of sham feeding-induced contraction. The results demonstrate that the cephalic phase of gallbladder contraction is probably mediated by M1 muscarinic receptors while the duodenal phase of gallbladder contraction is partially mediated by M1 receptors.


Multicomponent low molecular weight gelators.
Low molecular weight gelators (LMWG) self-assemble in solution into one-dimensional objects such as fibres or tapes. The entanglement of these fibres or tapes results in the formation of a network and a gel. In general, LMWG are investigated as single component systems. However, there are significant potential opportunities from mixed LMWG systems, which are rarely investigated. Here, we discuss the potential of multicomponent systems, and critically discuss the challenges.


Outcome of facial contour asymmetry after conventional two-dimensional versus computer-assisted three-dimensional planning in cleft orthognathic surgery.
Computer-assisted 3D planning has overcome the limitations of conventional 2D planning-guided orthognathic surgery (OGS), but difference for facial contour asymmetry outcome has not been verified to date. This comparative study assessed the facial contour asymmetry outcome of consecutive patients with unilateral cleft lip and palate who underwent 2D planning (n = 37)- or 3D simulation (n = 38)-guided OGS treatment for correction of maxillary hypoplasia and skeletal Class III malocclusion between 2010 and 2018. Normal age-, gender-, and ethnicity-matched individuals (n = 60) were enrolled for comparative analyses. 2D (n = 60, with 30 images for each group) and 3D (n = 43, with 18 and 25 images for 2D planning and 3D simulation groups, respectively) photogrammetric-based facial contour asymmetry-related measurements were collected from patients and normal individuals. The facial asymmetry was further verified by using subjective perception of a panel composed of 6 blinded raters. On average, the facial contour asymmetry was significantly (all p < 0.05) reduced after 3D virtual surgery planning for all tested parameters, with no significant differences between post-OGS 3D simulation-related values and normal individuals. No significant differences were observed for pre- and post-OGS values in conventional 2D planning-based treatment, with significant (all p < 0.05) differences for all normal individuals-related comparisons. This study suggests that 3D planning presents superior facial contour asymmetry outcome than 2D planning.


Insulin stimulates actin comet tails on intracellular GLUT4-containing compartments in differentiated 3T3L1 adipocytes.
Incubation of isolated GLUT4-containing vesicles with Xenopus oocyte extracts resulted in a guanosine 5'-[gamma-thio]triphosphate (GTP gamma S) and sodium orthovanadate stimulation of actin comet tails. The in vitro actin-based GLUT4 vesicle motility was inhibited by both latrunculin B and a dominant-interfering N-WASP mutant, N-WASP/Delta VCA. Preparations of gently sheared (broken) 3T3L1 adipocytes also displayed GTP gamma S and sodium orthovanadate stimulation of actin comet tails on GLUT4 intracellular compartments. Furthermore, insulin pretreatment of intact adipocytes prior to gently shearing also resulted in a marked increase in actin polymerization and actin comet tailing on GLUT4 vesicles. In addition, the insulin stimulation of actin comet tails was completely inhibited by Clostridum difficile toxin B, demonstrating a specific role for a Rho family member small GTP-binding protein. Expression of N-WASP/Delta VCA in intact cells had little effect on adipocyte cortical actin but partially inhibited insulin-stimulated GLUT4 translocation. Taken together, these data demonstrate that insulin can induce GLUT4 vesicle actin comet tails that are necessary for the efficient translocation of GLUT4 from intracellular storage sites to the plasma membrane.


Vaccine Hesitancy: In Search of the Risk Communication Comfort Zone.
This paper reports the findings of a national online survey to parents of children aged 5 and younger. The objectives of the study were to assess parental understanding of childhood immunizations, identify sources of information that they trust for vaccine-related content, assess where parents with young children stand on the key issues in the public debate about vaccination, and identify which risk communication messages are most effective for influencing the behaviours of vaccine hesitant parents. A total of 1,000 surveys (closed and open-ended questions) were administered in November 2015 using the Angus Reid Forum Panel, a key consumer panel consisting of approximately 150,000 Canadian adults aged 18 and older, spread across all geographic regions of Canada. Approximately 92% of the Canadian parents surveyed consider vaccines safe and effective, and trust doctors and public health officials to provide timely and credible vaccine-related information. However, a concerning number of them either believe or are uncertain whether there is a link between vaccines and autism (28%), worry that vaccines might seriously harm their children (27%), or believe the pharmaceutical industry is behind the push for mandatory immunization (33%). Moreover, despite the common assumption that social media are becoming the go-to source of health news and information, most parents still rely on traditional media and official government websites for timely and credible information about vaccines and vaccine preventable diseases, particularly during community-based disease outbreaks. Finally, parents reported high levels of support for pro-vaccine messaging that has been demonstrated in previous research to have little to no positive impact on behaviour change, and may even be counterproductive. The study's results are highly relevant in a context where public health officials are expending significant resources to increase rates of childhood immunization and combat vaccine hesitancy. The data offer insight into where parents stand on the political and public debate about mandatory vaccination, what aspects of vaccine science remain uncertain to them, which media and institutional sources they use and trust to navigate the health information environment, how they look for information and whom they trust during periods of health emergency or crisis, and which communication strategies are considered most effective in persuading vaccine hesitant parents to immunize their children.


5-hydroxytryptamine-stimulated accumulation of 1,2-diacylglycerol in the rabbit basilar artery: a role for protein kinase C in smooth muscle contraction.
1. 5-Hydroxytryptamine (5-HT) produced a concentration-dependent increase in the membrane concentration of 1,2-diacylglycerol (DG) in the rabbit isolated basilar artery, but did not stimulate the hydrolysis of membrane phosphoinositide. 2. The 5-HT-induced accumulation of DG could be blocked with the putative phospholipase C inhibitor 2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate (NCDC; 70 microM), but not with the protein kinase C inhibitor, 1-(5-isoquinolinesulphonyl)-2-methyl piperazine (H7; 50 microM). 3. Direct stimulation of protein kinase C with phorbol 12,13-dibutyrate (PDBu) produced sustained smooth muscle contraction which was fairly rapid in onset and could be reversed by H7 but not by NCDC. The inactive phorbol, 4 alpha phorbol 12,13-dideceonate, did not produce contraction in the basilar artery. 4. 5-HT-induced contractions (1 nM-100 microM) were blocked or greatly reduced in the presence of the protein kinase inhibitor H7 or polymyxin B, and with the phospholipase C inhibitor, NCDC. The concentrations of these inhibitors which abolished contraction to 5-HT, did not alter smooth muscle contraction produced in response to 30 mM K(+)-physiological salt solution (PSS). 5. These data suggest that DG production and the subsequent activation of PKC forms an important component of the cerebrovascular contractile response to 5-HT. As the DG does not appear to arise from membrane phosphatidylinositol, it appears that 5-HT can stimulate the production of this second messenger in cerebral arteries by a mechanism which is different from peripheral arteries.


Inhibition of RNA interference and modulation of transposable element expression by cell death in Drosophila.
RNA interference (RNAi) regulates gene expression by sequence-specific destruction of RNA. It acts as a defense mechanism against viruses and represses the expression of transposable elements (TEs) and some endogenous genes. We report that mutations and transgene constructs that condition cell death suppress RNA interference in adjacent cells in Drosophila melanogaster. The reversal of RNAi is effective for both the white (w) eye color gene and green fluorescent protein (GFP), indicating the generality of the inhibition. Antiapoptotic transgenes that reverse cell death will also reverse the inhibition of RNAi. Using GFP and a low level of cell death produced by a heat shock-head involution defective (hs-hid) transgene, the inhibition appears to occur by blocking the conversion of double-stranded RNA (dsRNA) to short interfering RNA (siRNA). We also demonstrate that the mus308 gene and endogenous transposable elements, which are both regularly silenced by RNAi, are increased in expression and accompanied by a reduced level of siRNA, when cell death occurs. The finding that chronic ectopic cell death affects RNAi is critical for an understanding of the application of the technique in basic and applied studies. These results also suggest that developmental perturbations, disease states, or environmental insults that cause ectopic cell death would alter transposon and gene expression patterns in the organism by the inhibition of small RNA silencing processes.


Rainfall simulation in greenhouse microcosms to assess bacterial-associated runoff from land-applied poultry litter.
Runoff water following a rain event is one possible source of environmental contamination after a manure application. This greenhouse study used a rainfall simulator to determine bacterial-associated runoff from troughs of common bermudagrass [Cynodon dactylon (L.) Pers.] that were treated with P-based, N-based, and N plus lime rates of poultry (Gallus gallus) litter, recommended inorganic fertilizer, and control. Total heterotrophic plate count (HPC) bacteria, total and thermotolerant coliforms, enterococci, staphylococci, Clostridium perfringens, Salmonella, and Campylobacter, as well as antibiotic resistance profiles for the staphylococci and enterococci isolates were all monitored in runoff waters. Analysis following five rainfall events indicated that staphylococci, enterococci, and clostridia levels were related to manure application rate. Runoff release of staphylococci, enterococci, and C. perfringens were approximately 3 to 6 log10 greater in litter vs. control treatment. In addition, traditional indicators such as thermotolerant and total coliforms performed poorly as fecal indicators. Some isolated enterococci demonstrated increased antibiotic resistance to polymixin b and/or select aminoglyocosides, while many staphylococci were susceptible to most antimicrobials tested. Results indicated poultry litter application can lead to microbial runoff following simulated rain events. Future studies should focus on the use of staphylococci, enterococci, and C. perfringens as indicators.


Magnolol inhibits Streptococcus suis-induced inflammation and ROS formation via TLR2/MAPK/NF-κB signaling in RAW264.7 cells.
Our previous studies have shown that Magnolol (Mag) improves the symptoms and decreases the levels of cytokines during infection induced by Streptococcus suis (S. suis) in mice. Although some reports show that Mag inhibits lipopolysaccharide-induced inflammatory responses via downregulating mitogen-activated protein kinases (MAPK) and nuclear factor-κB (NF-κB) signaling pathways, the molecular mechanisms underlying Mag-mediated inhibition of S. suis-induced inflammatory responses are poorly understood. Here, RAW264.7 cells were stimulated with S. suis in the presence or absence of Mag. Cell viability and bactericidal effects were examined, and the concentrations of tumor necrosis factor-a (TNF-α), IL-1β (interleukin-1β), IL-6 (interleukin-6), and IL-8 (interleukin- 8) were determined by ELISA. The change in ROS (reactive oxygen species) was determined by fluorescence microscopy and ELISA. The levels of Toll-like receptor 2 (TLR2) and MAPK family proteins and NF-κB signaling were determined by Western blot analysis. S. suis induced massive RAW264.7 cell death, a decline in bactericidal activity, the release of inflammatory cytokines, increased oxidative stress, and activation of TLR2/MAPK/NF-κB signaling pathways. Mag treatment significantly suppressed macrophage cell death and caused a decline in bactericidal activity. Furthermore, Mag decreased inflammatory cytokines production and ROS generation. It also prevented p38, extracellular regulated protein kinases (ERK), c-Jun N-terminal kinase (JNK), inhibitor of NF-κB (IκB), and NF-κB phosphorylation induced by S. suis in a dose-dependent manner. Our results indicate that Mag exerts anti-inflammatory and cell-protective effects and mediates the activation of MAPK and NF-κB signaling by downregulating the expression of TLR2 upregulated by S. suis.


Mechanical conditions that accelerate intervertebral disc degeneration: overload versus immobilization.
A review of the literature on macromechanical factors that accelerate disc degeneration with particular focus on distinguishing the roles of immobilization and overloading. This review examines evidence from the literature in the areas of biomechanics, epidemiology, animal models, and intervertebral disc physiology. The purpose is to examine: 1) what are the degeneration-related alterations in structural, material, and failure properties in the disc; and 2) evidence in the literature for causal relationships between mechanical loading and alterations in those structural and material properties that constitute disc degeneration. It is widely assumed that the mechanical environment of the intervertebral disc at least in part determines its rate of degeneration. However, there are two plausible and contrasting theories as to the mechanical conditions that promote degeneration: 1) mechanical overload; and 2) reduced motion and loading. There are a greater number of studies addressing the "wear and tear" theory than the immobilization theory. Evidence is accumulating to support the notion that there is a "safe window" of tissue mechanical conditions in which the discs remain healthy. It is concluded that probably any abnormal loading conditions (including overload and immobilization) can produce tissue trauma and/or adaptive changes that may result in disc degeneration. Adverse mechanical conditions can be due to external forces, or may result from impaired neuromuscular control of the paraspinal and abdominal muscles. Future studies will need to evaluate additional unquantified interactions between biomechanics and factors such as genetics and behavioral responses to pain and disability.


[Gender-specific differences in quality of life of patients with type 2 diabetes with or without participating in DMP].
Disease management programmes (DMP) are supposed to improve the care of patients with type 2 diabetes or other chronic conditions. One stated aim is the improvement of the health-related quality of life. Within the ELSID study (controlled study for the evaluation of the DMP for patients with type 2 diabetes) there has been a survey of insurants of the general regional health funds (AOK) by means of the SF-36. The aim of this survey is a comparison of patients participating in the Diabetes DMP with those who are not participating in the program with regard to their quality of life. A random sample of 3,546 patients with type 2 diabetes out of the total sample of the ELSID study (20,625) was asked to complete the SF-36. 1,532 questionnaires were returned (response rate 43.2%). 1,399 were analysed. Within all scales of the SF-36, men achieved higher scores than women. Differentiated in participants and non-participants in the DMP, in men there were lower scores for the participants than for the non-participants. In women this proportion was reversed. This cross-sectional-study provides first indications for gender-specific differences within the quality of life of patients with diabetes participating in the DMP compared to patients who are not participating. These differences should be considered more intensely within further research of DMP evaluation and the configuration of the programmes in the future.


Polarity of immunogens: implications for vaccine design.
Peptide constructs have been engineered consisting of amino acid sequence determinant recognized by T cells (TD) co-linearly linked to haptenic peptides. It was found that high anti-hapten antibody titers were induced after immunization with those constructs which had the TD sequence in the N-terminal position with respect to the hapten. Low or zero titers were elicited when the TD was in C-terminal position. Also, a high anti-hapten antibody titer corresponded to a low or zero anti-TD antibody titer and vice versa. These results suggest that immunogens are polar and stress the relevance of searching the more adequate position of the TD within a peptide construct when designing immunogens or synthetic peptide vaccines.


Effects of Glasgow Outcome Scale misclassification on traumatic brain injury clinical trials.
The Glasgow Outcome Scale (GOS) is the primary endpoint for efficacy analysis of clinical trials in traumatic brain injury (TBI). Accurate and consistent assessment of outcome after TBI is essential to the evaluation of treatment results, particularly in the context of multicenter studies and trials. The inconsistent measurement or interobserver variation on GOS outcome, or for that matter, on any outcome scales, may adversely affect the sensitivity to detect treatment effects in clinical trial. The objective of this study is to examine effects of nondifferential misclassification of the widely used five-category GOS outcome scale and in particular to assess the impact of this misclassification on detecting a treatment effect and statistical power. We followed two approaches. First, outcome differences were analyzed before and after correction for misclassification using a dataset of 860 patients with severe brain injury randomly sampled from two TBI trials with known differences in outcome. Second, the effects of misclassification on outcome distribution and statistical power were analyzed in simulation studies on a hypothetical 800-patient dataset. Three potential patterns of nondifferential misclassification (random, upward and downward) on the dichotomous GOS outcome were analyzed, and the power of finding treatments differences was investigated in detail. All three patterns of misclassification reduce the power of detecting the true treatment effect and therefore lead to a reduced estimation of the true efficacy. The magnitude of such influence not only depends on the size of the misclassification, but also on the magnitude of the treatment effect. In conclusion, nondifferential misclassification directly reduces the power of finding the true treatment effect. An awareness of this procedural error and methods to reduce misclassification should be incorporated in TBI clinical trials.


Raman spectroscopy adds complementary detail to the high-resolution x-ray crystal structure of photosynthetic PsbP from Spinacia oleracea.
Raman microscopy permits structural analysis of protein crystals in situ in hanging drops, allowing for comparison with Raman measurements in solution. Nevertheless, the two methods sometimes reveal subtle differences in structure that are often ascribed to the water layer surrounding the protein. The novel method of drop-coating deposition Raman spectropscopy (DCDR) exploits an intermediate phase that, although nominally "dry," has been shown to preserve protein structural features present in solution. The potential of this new approach to bridge the structural gap between proteins in solution and in crystals is explored here with extrinsic protein PsbP of photosystem II from Spinacia oleracea. In the high-resolution (1.98 Å) x-ray crystal structure of PsbP reported here, several segments of the protein chain are present but unresolved. Analysis of the three kinds of Raman spectra of PsbP suggests that most of the subtle differences can indeed be attributed to the water envelope, which is shown here to have a similar Raman intensity in glassy and crystal states. Using molecular dynamics simulations cross-validated by Raman solution data, two unresolved segments of the PsbP crystal structure were modeled as loops, and the amino terminus was inferred to contain an additional beta segment. The complete PsbP structure was compared with that of the PsbP-like protein CyanoP, which plays a more peripheral role in photosystem II function. The comparison suggests possible interaction surfaces of PsbP with higher-plant photosystem II. This work provides the first complete structural picture of this key protein, and it represents the first systematic comparison of Raman data from solution, glassy, and crystalline states of a protein.


High genetic diversity and adaptive potential of two simian hemorrhagic fever viruses in a wild primate population.
Key biological properties such as high genetic diversity and high evolutionary rate enhance the potential of certain RNA viruses to adapt and emerge. Identifying viruses with these properties in their natural hosts could dramatically improve disease forecasting and surveillance. Recently, we discovered two novel members of the viral family Arteriviridae: simian hemorrhagic fever virus (SHFV)-krc1 and SHFV-krc2, infecting a single wild red colobus (Procolobus rufomitratus tephrosceles) in Kibale National Park, Uganda. Nearly nothing is known about the biological properties of SHFVs in nature, although the SHFV type strain, SHFV-LVR, has caused devastating outbreaks of viral hemorrhagic fever in captive macaques. Here we detected SHFV-krc1 and SHFV-krc2 in 40% and 47% of 60 wild red colobus tested, respectively. We found viral loads in excess of 10(6)-10(7) RNA copies per milliliter of blood plasma for each of these viruses. SHFV-krc1 and SHFV-krc2 also showed high genetic diversity at both the inter- and intra-host levels. Analyses of synonymous and non-synonymous nucleotide diversity across viral genomes revealed patterns suggestive of positive selection in SHFV open reading frames (ORF) 5 (SHFV-krc2 only) and 7 (SHFV-krc1 and SHFV-krc2). Thus, these viruses share several important properties with some of the most rapidly evolving, emergent RNA viruses.


Information-seeking about health in a community sample of adults: correlates and associations with other health-related practices.
Action by individuals to acquire information about their health has been an element incorporated throughout theory, research, and programs related to health promotion. This report describes an attempt to determine if an information-seeking dimension could be empirically identified in a general community-resident sample, and if so, to examine some of its characteristics. A total of 281 adults aged 18-75 were contacted by telephone using random digit dialing and were interviewed about a variety of personal health practices. Factor analysis identified a five-item cluster representing a tendency to seek out information about health. Women were more likely than men to report seeking information. In addition, more frequent information-seeking was associated with favorable responses to several other health-related practices. Formal health service use was the only type of health practice not associated with information-seeking, perhaps because regularity of contact is influenced strongly by health professionals (e.g., reminder cards and having staff call to schedule annual exams). Overall, results of the investigation support the importance of information-seeking as a component of a personal health practice repertoire. Additional attention might be directed toward elaborating its role as a "process" variable in health education programs and social marketing efforts, particularly in areas such as response to recruitment messages, dropout vs. maintenance, and differential gains on outcome measures of program effectiveness.


Identification and antigenic typing of Rickettsia tsutsugamushi in naturally infected chiggers (Acarina: Trombiculidae) by direct immunofluorescence.
Rickettsia tsutsugamushi organisms were detected and typed antigenically by direct immunofluorescence in mites from laboratory-maintained infected colonies of Leptotrombidium (Leptotrombidium) fletcheri and L. (L.) arenicola. Rickettsiae were identified most readily in unengorged larvae, but were also discernable in engorged larvae and all post-larval stages of the vectors.


Sensory Processing in the General Population: Adaptability, Resiliency, and Challenging Behavior.
Sensory processing is a personal trait that can contribute to challenging behavior. Protective factors, such as resiliency, can support children in managing their behavior, yet little research has addressed the contribution of sensory processing to protective factors. Therefore, in this cross-sectional study we aimed to determine the contribution of sensory processing (using the Sensory Profile-2) to challenging behavior and protective factors (using the Behavior Assessment System for Children, 2nd ed., Parent Rating Scales) in a sample of 51 children ages 6-11 yr from the general U.S. The results indicated that sensory avoiding predicts externalizing behaviors, depression, resiliency, and adaptability. In addition, sensory seeking is related to depression and resiliency, whereas sensory sensitivity is related to externalizing behaviors. Implications for occupational therapy research and practice are discussed.


Development of functional chest imaging with a dynamic flat-panel detector (FPD).
Dynamic FPD permits the acquisition of distortion-free radiographs with a large field of view and high image quality. In the present study, we investigated the feasibility of functional imaging for evaluating the pulmonary sequential blood distribution with an FPD, based on changes in pixel values during cardiac pumping. Dynamic chest radiographs of seven normal subjects were obtained in the expiratory phase by use of an FPD system. We measured the average pixel value in each region of interest that was located manually in the heart and lung areas. Subsequently, inter-frame differences and differences from a minimum-intensity projection image, which was created from one cardiac cycle, were calculated. These difference values were then superimposed on dynamic chest radiographs in the form of a color display, and sequential blood distribution images and a blood distribution map were created. The results were compared to typical data on normal cardiac physiology. The clinical effectiveness of our method was evaluated in a patient who had abnormal pulmonary blood flow. In normal cases, there was a strong correlation between the cardiac cycle and changes in pixel value. Sequential blood distribution images showed a normal pattern at determined by the physiology of pulmonary blood flow, with a symmetric distribution and no blood flow defects throughout the entire lung region. These findings indicated that pulmonary blood flow was reflected on dynamic chest radiographs. In an abnormal case, a defect in blood flow was shown as defective in color in a blood distribution map. The present method has the potential for evaluation of local blood flow as an optional application in general chest radiography.


Relatively rare root endophytic bacteria drive plant resource allocation patterns and tissue nutrient concentration in unpredictable ways.
Plant endophytic bacterial strains can influence plant traits such as leaf area and root length. Yet, the influence of more complex bacterial communities in regulating overall plant phenotype is less explored. Here, in two complementary experiments, we tested whether we can predict plant phenotype response to changes in microbial community composition. In the first study, we inoculated a single genotype of Populus deltoides with individual root endophytic bacteria and measured plant phenotype. Next, data from this single inoculation were used to predict phenotypic traits after mixed three-strain community inoculations, which we tested in the second experiment. By itself, each bacterial endophyte significantly but weakly altered plant phenotype relative to noninoculated plants. In a mixture, bacterial strain Burkholderia BT03, constituted at least 98% of community relative abundance. Yet, plant resource allocation and tissue nutrient concentrations were disproportionately influenced by Pseudomonas sp. GM17, GM30, and GM41. We found a 10% increase in leaf mass fraction and an 11% decrease in root mass fraction when replacing Pseudomonas GM17 with GM41 in communities containing both Pseudomonas GM30 and Burkholderia BT03. Our results indicate that interactions among endophytic bacteria may drive plant phenotype over the contribution of each strain individually. Additionally, we have shown that low-abundance strains contribute to plant phenotype challenging the assumption that the dominant strains will drive plant function.


Parallel pathways from motor and somatosensory cortex for controlling whisker movements in mice.
Mice can gather tactile sensory information by actively moving their whiskers to palpate objects in their immediate surroundings. Whisker sensory perception therefore requires integration of sensory and motor information, which occurs prominently in the neocortex. The signalling pathways from the neocortex for controlling whisker movements are currently poorly understood in mice. Here, we delineate two pathways, one originating from primary whisker somatosensory cortex (wS1) and the other from whisker motor cortex (wM1), that control qualitatively distinct movements of contralateral whiskers. Optogenetic stimulation of wS1 drove retraction of contralateral whiskers while stimulation of wM1 drove rhythmic whisker protraction. To map brainstem pathways connecting these cortical areas to whisker motor neurons, we used a combination of anterograde tracing using adenoassociated virus injected into neocortex and retrograde tracing using monosynaptic rabies virus injected into whisker muscles. Our data are consistent with wS1 driving whisker retraction by exciting glutamatergic premotor neurons in the rostral spinal trigeminal interpolaris nucleus, which in turn activate the motor neurons innervating the extrinsic retractor muscle nasolabialis. The rhythmic whisker protraction evoked by wM1 stimulation might be driven by excitation of excitatory and inhibitory premotor neurons in the brainstem reticular formation innervating both intrinsic and extrinsic muscles. Our data therefore begin to unravel the neuronal circuits linking the neocortex to whisker motor neurons.


Tighten after Relax: Minimax-Optimal Sparse PCA in Polynomial Time.
We provide statistical and computational analysis of sparse Principal Component Analysis (PCA) in high dimensions. The sparse PCA problem is highly nonconvex in nature. Consequently, though its global solution attains the optimal statistical rate of convergence, such solution is computationally intractable to obtain. Meanwhile, although its convex relaxations are tractable to compute, they yield estimators with suboptimal statistical rates of convergence. On the other hand, existing nonconvex optimization procedures, such as greedy methods, lack statistical guarantees. In this paper, we propose a two-stage sparse PCA procedure that attains the optimal principal subspace estimator in polynomial time. The main stage employs a novel algorithm named sparse orthogonal iteration pursuit, which iteratively solves the underlying nonconvex problem. However, our analysis shows that this algorithm only has desired computational and statistical guarantees within a restricted region, namely the basin of attraction. To obtain the desired initial estimator that falls into this region, we solve a convex formulation of sparse PCA with early stopping. Under an integrated analytic framework, we simultaneously characterize the computational and statistical performance of this two-stage procedure. Computationally, our procedure converges at the rate of [Formula: see text] within the initialization stage, and at a geometric rate within the main stage. Statistically, the final principal subspace estimator achieves the minimax-optimal statistical rate of convergence with respect to the sparsity level s*, dimension d and sample size n. Our procedure motivates a general paradigm of tackling nonconvex statistical learning problems with provable statistical guarantees.


Using graphene nano-particle embedded in photonic crystal fiber for evanescent wave mode-locking of fiber laser.
A photonic crystal fiber (PCF) with high-quality graphene nano-particles uniformly dispersed in the hole cladding are demonstrated to passively mode-lock the erbium-doped fiber laser (EDFL) by evanescent-wave interaction. The few-layer graphene nano-particles are obtained by a stabilized electrochemical exfoliation at a threshold bias. These slowly and softly exfoliated graphene nano-particle exhibits an intense 2D band and an almost disappeared D band in the Raman scattering spectrum. The saturable phenomena of the extinction coefficient β in the cladding provides a loss modulation for the intracavity photon intensity by the evanescent-wave interaction. The evanescent-wave mode-locking scheme effectively enlarges the interaction length of saturable absorption with graphene nano-particle to provide an increasing transmittance ΔT of 5% and modulation depth of 13%. By comparing the core-wave and evanescent-wave mode-locking under the same linear transmittance, the transmittance of the graphene nano-particles on the end-face of SMF only enlarges from 0.54 to 0.578 with ΔT = 3.8% and the modulation depth of 10.8%. The evanescent wave interaction is found to be better than the traditional approach which confines the graphene nano-particles at the interface of two SMF patchcords. When enlarging the intra-cavity gain by simultaneously increasing the pumping current of 980-nm and 1480-nm pumping laser diodes (LDs) to 900 mA, the passively mode-locked EDFL shortens its pulsewidth to 650 fs and broadens its spectral linewidth to 3.92 nm. An extremely low carrier amplitude jitter (CAJ) of 1.2-1.6% is observed to confirm the stable EDFL pulse-train with the cladding graphene nano-particle based evanescent-wave mode-locking.


Effects of an anti-smoking program to prevent lung cancer among urban aboriginals in Taiwan.
Indigenous people who leave their hometowns and move to the city to earn a living became urban aboriginals. During the process of adapting to urban living situations, they may use various coping strategies such as smoking to overcome their stress. Therefore, it is crucial to provide health education including smoking prevention, increasing knowledge regarding of tobacco hazard, self-efficacy of anti-smoking, and adjusting smoking behavior so as to empower their anti-smoking motivation to prevent lung cancer. The purpose of this study was to explore the effectiveness of an anti-smoking program on urban aboriginals in Taiwan. A quasi-experimental study design with purposeful sampling was employed. A total of 125 aboriginal subjects were recruited from two local churches at Shu Lin area in northern Taiwan. Subjects were divided into an experimental group (n =64 ) and a control group (n = 61). Both took pre-tests in order to set baseline values, and only the experimental group participated for 3-weeks in the anti-smoking program classes. Both groups took post-tests immediately after the intervention in order to evaluate the immediate effects of the teaching program, and a follow-up test was conducted four weeks after the intervention. Data were analyzed using descriptive statistics, one-way ANCOVA, and repeat measure ANCOVA. After controlling for confounding variables, the results showed that there were statistically significant differences in the self-efficacy of anti-smoking and smoking behavior between experimental and control groups in the immediately post-test and the follow-up test (p < 0.05). However, there was no significant differences in the recognition of hazards of smoking at eiter time point. The findings of this study revealed that the anti-smoking program effectively improved self-efficacy of anti-smoking, and decreased the smoking behavior in urban aboriginals. They provide useful information as a reference regarding of aboriginal health promotion to health providers. It is imperative that anti-smoking be reinforced for those regular smokers to prevent induction of lung cancer.


Activation of mouse macrophages for tumor cell killing. I. Quantitative analysis of interactions between lymphokine and lipopolysaccharide.
Lymphokine-rich supernatants from concanavalin A-stimulated spleen cell cultures failed to activate mouse peritoneal macrophages for tumor cell killing, providing that the assay system was free of detectable (less than 0.125 ng/ml) endotoxin. Instead, increasing amounts of lymphokine progressively increased the sensitivity of macrophages to activation by purified bacterial lipopolysaccharide (LPS). Conversely, the effect of minute amounts of lymphokine could be detected in the presence of relatively high (though nonactivating) concentrations of LPS. The latter observation was exploited to develop a highly sensitive, quantitative assay for lymphokine. In this assay, serially diluted supernatants were tested on macrophage monolayers in the presence of a constant concentration (3 ng/ml) of LPS. Under these conditions, the lymphokine dose-response curve was sigmoidal and therefore suitable for conversion to a linear plot using the logarithmic transformation of the von Krogh equation. Activity could be expressed quantitatively and reproducibly in terms of units of activity. By facilitating purification the assay should contribute to the development of a more precise biochemical understanding of the role of lymphokine in macrophage activation for tumor cell killing.


Optical pulse propagation in the tight-binding approximation.
We formulate the equations describing pulse propagation in a one-dimensional optical structure described by the tight binding approximation, commonly used in solid-state physics to describe electrons levels in a periodic potential. The analysis is carried out in a way that highlights the correspondence with the analysis of pulse propagation in a conventional waveguide. Explicit expressions for the pulse in the waveguide are derived and discussed in the context of the sampling theorems of finite-energy space and time signals.


Automated analysis of the 12-lead ECG in the emergency department: association between high-sensitivity cardiac troponin I and the cardiac electrical biomarker.
Timely detection of myocardial injury is essential for appropriate management of patients in emergency department (ED) evaluated for acute myocardial infarction. A novel electrocardiogram (ECG) metric, the Cardiac Electrical Biomarker (CEB), uses eigenvalue modeling of the 12-lead ECG and quantifies dipolar vs. multipolar forces. The goal of this project was to study association between the CEB and high-sensitivity troponin I (HsTnI). We conducted a retrospective study of patients, evaluated in the ED for acute myocardial infarction [n = 411; 57.6 ± 13.2 years; 186 (45%) men; 266 (64%) African-Americans]. Resting 12-lead ECG and HsTnI were measured at presentation and at 3, 6, and 9 hours after the initial measurement. The CEB was measured by the VectraplexECG System (VectraCor, Totowa, NJ). Patient-specific longitudinal analysis was performed to study association between the CEB with HsTnI changes over time. The CEB indicated myocardial injury in 116 (28.2%) study participants. HsTnI was significantly elevated during ED observation period in patients with myocardial injury, diagnosed by the CEB [median (interquartile range), 10.3 (5.2-31.4) vs. 6.3 (3.5-16.5) ng/L; P = 0.002]. In a mixed-effects linear regression adjusted for age, race, and sex, increasing HsTnI was associated with the CEB elevation [β-coefficient, 0.071 (95% confidence interval, 0.008-0.134); P = 0.027]. In conclusion, in patients in ED evaluated for acute myocardial injury, increasing values of HsTnI were associated with increasing values of the CEB, suggesting that myocardial injury is the mechanism that underlines acute changes in the CEB.


Fatal venous thromboembolism after splenectomy: pathogenesis and management.
Venous thromboembolism (VTE) occurs in approximately 10% of patients after splenectomy, particularly splenectomy performed for hematologic abnormalities. Clinicians often fail to recognize this potential complication in the postoperative period, leading to inappropriate anticoagulation prophylaxis and treatment for these patients. The authors discuss the pathophysiologic mechanisms of VTE in patients who undergo splenectomy and offer management strategies for this complication. A case report of a patient who underwent splenectomy for idiopathic thrombocytopenia purpura, with subsequent fatal VTE, highlights the importance of this issue. The authors also review current guidelines for managing venous thromboprophylaxis in patients who undergo general, laparoscopic, and cancer-related surgical procedures, and they compare these patients to those who undergo splenectomy.


Sample size re-estimation in paired comparative diagnostic accuracy studies with a binary response.
The sample size required to power a study to a nominal level in a paired comparative diagnostic accuracy study, i.e. studies in which the diagnostic accuracy of two testing procedures is compared relative to a gold standard, depends on the conditional dependence between the two tests - the lower the dependence the greater the sample size required. A priori, we usually do not know the dependence between the two tests and thus cannot determine the exact sample size required. One option is to use the implied sample size for the maximal negative dependence, giving the largest possible sample size. However, this is potentially wasteful of resources and unnecessarily burdensome on study participants as the study is likely to be overpowered. A more accurate estimate of the sample size can be determined at a planned interim analysis point where the sample size is re-estimated. This paper discusses a sample size estimation and re-estimation method based on the maximum likelihood estimates, under an implied multinomial model, of the observed values of conditional dependence between the two tests and, if required, prevalence, at a planned interim. The method is illustrated by comparing the accuracy of two procedures for the detection of pancreatic cancer, one procedure using the standard battery of tests, and the other using the standard battery with the addition of a PET/CT scan all relative to the gold standard of a cell biopsy. Simulation of the proposed method illustrates its robustness under various conditions. The results show that the type I error rate of the overall experiment is stable using our suggested method and that the type II error rate is close to or above nominal. Furthermore, the instances in which the type II error rate is above nominal are in the situations where the lowest sample size is required, meaning a lower impact on the actual number of participants recruited. We recommend multinomial model maximum likelihood estimation of the conditional dependence between paired diagnostic accuracy tests at an interim to reduce the number of participants required to power the study to at least the nominal level. ISRCTN ISRCTN73852054 . Registered 9th of January 2015. Retrospectively registered.


Novel preparation of controlled porosity particle/fibre loaded scaffolds using a hybrid micro-fluidic and electrohydrodynamic technique.
The purpose of this research was to produce multi-dimensional scaffolds containing biocompatible particles and fibres. To achieve this, two techniques were combined and used: T-Junction microfluidics and electrohydrodynamic (EHD) processing. The former was used to form layers of monodispersed bovine serum albumin (BSA) bubbles, which upon drying formed porous scaffolds. By altering the T-Junction processing parameters, bubbles with different diameters were produced and hence the scaffold porosity could be controlled. EHD processing was used to spray or spin poly(lactic-co-glycolic) (PLGA), polymethysilsesquioxane (PMSQ) and collagen particles/fibres onto the scaffolds during their production and after drying. As a result, multifunctional BSA scaffolds with controlled porosity containing PLGA, PMSQ and collagen particles/fibres were obtained. Product morphology was studied by optical and scanning electron microscopy. These products have potential applications in many advanced biomedical, pharmaceutical and cosmetic fields e.g. bone regeneration, drug delivery, cosmetic cream lathers, facial scrubbing creams etc.


Can novel genetic analyses help to identify low-dispersal marine invasive species?
Genetic methods can be a powerful tool to resolve the native versus introduced status of populations whose taxonomy and biogeography are poorly understood. The genetic study of introduced species is presently dominated by analyses that identify signatures of recent colonization by means of summary statistics. Unfortunately, such approaches cannot be used in low-dispersal species, in which recently established populations originating from elsewhere in the species' native range also experience periods of low population size because they are founded by few individuals. We tested whether coalescent-based molecular analyses that provide detailed information about demographic history supported the hypothesis that a sea squirt whose distribution is centered on Tasmania was recently introduced to mainland Australia and New Zealand through human activities. Methods comparing trends in population size (Bayesian Skyline Plots and Approximate Bayesian Computation) were no more informative than summary statistics, likely because of recent intra-Tasmanian dispersal. However, IMa2 estimates of divergence between putatively native and introduced populations provided information at a temporal scale suitable to differentiate between recent (potentially anthropogenic) introductions and ancient divergence, and indicated that all three non-Tasmanian populations were founded during the period of European settlement. While this approach can be affected by inaccurate molecular dating, it has considerable (albeit largely unexplored) potential to corroborate nongenetic information in species with limited dispersal capabilities.


Arterial baroreflex control of sympathetic nerve activity during elevation of blood pressure in normal man: dominance of aortic baroreflexes.
Arterial baroreceptors in the carotid sinus (CBR) and aortic arch (ABR) regions exert important control over heart rate and peripheral vascular responses to changes in arterial pressure. The relative roles of these two baroreflex pathways on control of sympathetic nerve activity during sustained elevation of arterial pressure in man is unknown. We therefore studied the relative contributions of the carotid versus the aortic baroreflexes on the control of muscle sympathetic nerve activity (MSNA) during elevation of arterial pressure in normal human subjects. In eight normal men (group I), we measured MSNA (microneurography) during sustained elevation of arterial pressure produced by intravenous infusion of phenylephrine (PE) alone (combined ABR and CBR activation) versus during PE infusion with superimposed application of sustained external neck pressure (NP). NP was applied during sustained PE infusion to eliminate the increase in transmural carotid sinus pressure and thus remove CBR activation, thereby causing ABR stimulation alone. Mean arterial pressure was measured directly, central venous pressure was held constant during PE infusion, and MSNA was measured as total activity (burst frequency X amplitude) and expressed as units. Infusion of PE (ABR and CBR activation) increased mean arterial pressure from 87.2 +/- 2.8 to 94.9 +/- 2.9 mm Hg (+/- SE, p less than .001). This was accompanied by a decrease in heart rate from 65.8 +/- 3.4 to 56.1 +/- 3.3 beats/min (p less than .001) and a decrease in MSNA from 236.2 +/- 47.5 to 84.5 +/- 19.3 units (p less than .001). During infusion of PE with superimposed NP (ABR activation alone), mean arterial pressure increased further to 101.2 +/- 2.9 mm Hg (p less than .001 versus control or PE alone), and heart rate returned to control levels of 62.9 +/- 2.0 beats/min (p = NS vs control; p less than .01 PE vs PE plus NP), but MSNA remained reduced at 48.6 +/- 9.2 units (p less than .01 vs control; p = NS vs PE alone). Thus, combined activation of ABR and CBR resulted in a 65 +/- 5% attenution of MSNA, while activation of ABR alone resulted in a 73 +/- 7% attenuation of MSNA. In a separate series of experiments in seven subjects (group II) we used sustained external neck suction alone to activate the CBR (leaving the ABR either unchanged or minimally deactivated) and studied the MSNA responses to this CBR activation.(ABSTRACT TRUNCATED AT 400 WORDS)


Distributed brain co-processor for tracking spikes, seizures and behaviour during electrical brain stimulation.
Early implantable epilepsy therapy devices provided open-loop electrical stimulation without brain sensing, computing, or an interface for synchronized behavioural inputs from patients. Recent epilepsy stimulation devices provide brain sensing but have not yet developed analytics for accurately tracking and quantifying behaviour and seizures. Here we describe a distributed brain co-processor providing an intuitive bi-directional interface between patient, implanted neural stimulation and sensing device, and local and distributed computing resources. Automated analysis of continuous streaming electrophysiology is synchronized with patient reports using a handheld device and integrated with distributed cloud computing resources for quantifying seizures, interictal epileptiform spikes and patient symptoms during therapeutic electrical brain stimulation. The classification algorithms for interictal epileptiform spikes and seizures were developed and parameterized using long-term ambulatory data from nine humans and eight canines with epilepsy, and then implemented prospectively in out-of-sample testing in two pet canines and four humans with drug-resistant epilepsy living in their natural environments. Accurate seizure diaries are needed as the primary clinical outcome measure of epilepsy therapy and to guide brain-stimulation optimization. The brain co-processor system described here enables tracking interictal epileptiform spikes, seizures and correlation with patient behavioural reports. In the future, correlation of spikes and seizures with behaviour will allow more detailed investigation of the clinical impact of spikes and seizures on patients.


Association between pregnancy at enrollment into HIV care and loss to care among women in the Democratic Republic of Congo, 2006-2013.
Loss to care is high among asymptomatic HIV-infected women initiated on antiretroviral therapy (ART) during pregnancy or in the postpartum period. However, whether pregnancy itself plays a role in the high loss to care rate is uncertain. We compared loss to care over seven years between pregnant and non-pregnant women at enrollment into HIV care in the Democratic Republic of Congo (DRC). We conducted a retrospective analysis of all ART-naive women aged 15-45 initiating HIV care at two large clinics in Kinshasa, DRC, from 2007-2013. Pregnancy status was recorded at care enrollment. Patients were classified as having no follow-up if they did not return to care after the initial enrollment visit. Among those with at least one follow-up visit after enrollment, we classified patients as lost to care if more than 365 days had passed since their last clinic visit. We used logistic regression to model the association between pregnancy status and no follow-up, and Cox proportional hazards regression to model the association between pregnancy status and time to loss to care. Of 2175 women included in the analysis, 1497 (68.8%) were pregnant at enrollment. Compared to non-pregnant women, pregnant women were less likely to be over 35 years of age (19.1% vs. 31.9%, p<0.0001) and less likely to be in WHO stage III or IV (9.0% vs. 26.3%, p<0.0001). Among pregnant women, 106 (7.1%) were not seen after enrollment, versus 25 (3.7%) non-pregnant women (adjusted odds ratio 2.01, 95% CI 1.24-3.24). Of the 2,044 women with at least one follow-up visit, 46.5% of pregnant women and 46.7% of non-pregnant women were lost to care by 5 years; hazards of loss to care were similar for pregnant and non-pregnant women (adjusted hazard ratio 1.08, 95% CI 0.93-1.26). In this large cohort of HIV-infected women, patients pregnant at care enrollment were more likely to never return for follow-up. Among those who attended at least one follow-up visit, loss to care was not different between pregnant and non-pregnant women, suggesting that pregnancy itself may not be the main driver of the high attrition observed in this cohort.


A comparison of statistical methods for identifying out-of-date systematic reviews.
Systematic reviews (SRs) can provide accurate and reliable evidence, typically about the effectiveness of health interventions. Evidence is dynamic, and if SRs are out-of-date this information may not be useful; it may even be harmful. This study aimed to compare five statistical methods to identify out-of-date SRs. A retrospective cohort of SRs registered in the Cochrane Pregnancy and Childbirth Group (CPCG), published between 2008 and 2010, were considered for inclusion. For each eligible CPCG review, data were extracted and "3-years previous" meta-analyses were assessed for the need to update, given the data from the most recent 3 years. Each of the five statistical methods was used, with random effects analyses throughout the study. Eighty reviews were included in this study; most were in the area of induction of labour. The numbers of reviews identified as being out-of-date using the Ottawa, recursive cumulative meta-analysis (CMA), and Barrowman methods were 34, 7, and 7 respectively. No reviews were identified as being out-of-date using the simulation-based power method, or the CMA for sufficiency and stability method. The overall agreement among the three discriminating statistical methods was slight (Kappa = 0.14; 95% CI 0.05 to 0.23). The recursive cumulative meta-analysis, Ottawa, and Barrowman methods were practical according to the study criteria. Our study shows that three practical statistical methods could be applied to examine the need to update SRs.


Reprint of: The regulation project: Tools for engaging the public in the legal regulation of drugs.
Lack of legal regulation and oversight of scheduled drugs in Canada has led to an unsafe drug supply responsible for the deaths of tens of thousands. In addition to contributing to the worst unregulated drug poisoning crisis in Canada's history, the policy framework prohibiting non-medical access to certain drugs has exacerbated numerous public health and safety concerns. An alternative approach to prohibition is for government to retake control of the currently illegal drug market through legal regulatory mechanisms. This paper presents the work of an ongoing international collaboration of organizations advocating legal regulation and some of the knowledge translation tools used to educate and engage the public on legal regulation within Canada. In order to encourage thinking and decision-making among stakeholders in a productive way, models of legal regulation for various substances were created as discussion tools to emerge values and considerations supporting different approaches. The models focus on five questions: 1) who has access to drugs; 2) how access is obtained; 3) where drugs can be accessed; 4) how much people can obtain; and 5) where drugs can be consumed. The models were presented to stakeholders in the context of an international meeting on legal regulation, and then adapted to a more "user-friendly" form: a collaborative negotiation-based Regulation Game, which was presented at a workshop in Montréal, Canada. Engaging different stakeholder groups on policy choices of legal regulation revealed initial barriers that we feel more confident can be overcome through creative and innovative tools such as the Regulation Game. Use of the game as a foundation for more traditional focus groups could be effective in reducing barriers to fulsome policy discussions on legal regulation.


Optical Coherence Tomography Angiography of Retinal Microvascular Changes Overlying Choroidal Nodules in Neurofibromatosis Type 1.
To report 3 cases of neurofibromatosis type 1 (NF1) with choroidal nodules associated with retinal microvascular changes imaged with optical coherence tomography angiography (OCTA). Small case series in 3 NF1 patients. OCTA examinations were performed by a trained examiner (J.J.) after pupillary dilation. A standard scan, centered over the macula measuring 6 × 6 mm and 3 × 3 mm was obtained according to the findings on standard color photography. Additional scans were obtained in the zones with microvascular abnormalities. The segmentation provided by the machine software was used. Corkscrew retinal vessels were observed in association with "placoid"-type choroidal nodules as shown by near-infrared reflectance imaging. In all cases, multiple lesions were found. They were second- or third-order tortuous vessels originating from the superior or inferior temporal veins. OCTA demonstrated that the tortuous venules were located in the superficial capillary plexus, and no abnormalities were found in the deep capillary plexus. Corkscrew retinal vessels are part of a spectrum of retinal microvascular alterations seen in association, sometimes overlying choroidal nodules in patients with NF1 and are visualized in the superficial capillary plexus on OCTA. We demonstrated with OCTA that they are not associated with flow loss or ischemia in the superficial and deep capillary plexus. The link between the underlying nodule remains unclear. Since neovascularization was described in choroidal ganglioneuroma, we hypothesize that corresponding secretory substances from Schwann cells, ganglion cells, or melanocytes in choroidal nodules might alter the retinal vasculature. We report on 3 cases of NF1 with choroidal nodules in association with retinal microvascular changes imaged with OCTA. OCTA demonstrated preservation of the blood flow in the deep and superficial capillary plexus of the retina. We hypothesize that angiogenic factors secreted by the underlying choroidal nodules could have an effect on the retinal vasculature. Further immunohistological studies in NF1 patients with choroidal nodules to detect angiogenic factors (such as VEGF) are necessary to confirm this hypothesis.