Reversal of radiation resistance in LNCaP cells by targeting apoptosis through ceramide synthase.
Cell lines derived from human prostate cancer are regarded as relatively resistant to both radiation-induced clonogenic death and apoptosis. Here we attempted to modulate the response of LNCaP prostate cancer cells to radiation therapy (XRT) by pretreatment with 12-O-tetradecanoylphorbol acetate (TPA), a known apoptogenic agent in LNCaP cells. Using plateau-phase cultures, we investigated the response of these cells to XRT, TPA, and a combination of XRT and TPA. LNCaP irradiation did not result in ceramide generation or apoptosis. However, pretreatment with TPA enabled XRT to generate ceramide via activation of the enzyme ceramide synthase and signal apoptosis. Apoptosis was abrogated by the competitive inhibitor of ceramide synthase, fumonisin B1. Furthermore, when transplanted orthotopically into the prostate of nude mice, LNCaP cells produced tumors that recapitulated the responses of LNCaP cells in vitro. XRT or TPA failed to signal apoptosis in LNCaP tumors, whereas a combination of the two resulted in substantial (20-25%) apoptosis within 24 h. There was an additional benefit associated with this regimen because TPA pretreatment protected the adjacent rectum from radiation-induced apoptosis. This represents the first description of signaling-based therapy designed to overcome one form of radiation resistance expressed preferentially in LNCaP human prostate cancer cells.


Humans thrived in South Africa through the Toba eruption about 74,000 years ago.
Approximately 74 thousand years ago (ka), the Toba caldera erupted in Sumatra. Since the magnitude of this eruption was first established, its effects on climate, environment and humans have been debated. Here we describe the discovery of microscopic glass shards characteristic of the Youngest Toba Tuff-ashfall from the Toba eruption-in two archaeological sites on the south coast of South Africa, a region in which there is evidence for early human behavioural complexity. An independently derived dating model supports a date of approximately 74 ka for the sediments containing the Youngest Toba Tuff glass shards. By defining the input of shards at both sites, which are located nine kilometres apart, we are able to establish a close temporal correlation between them. Our high-resolution excavation and sampling technique enable exact comparisons between the input of Youngest Toba Tuff glass shards and the evidence for human occupation. Humans in this region thrived through the Toba event and the ensuing full glacial conditions, perhaps as a combined result of the uniquely rich resource base of the region and fully evolved modern human adaptation.


Genetic variation and population structure of Botswana populations as identified with AmpFLSTR Identifiler short tandem repeat (STR) loci.
Population structure was investigated in 990 Botswana individuals according to ethno-linguistics, Bantu and Khoisan, and geography (the nine administrative districts) using the Identifiler autosomal microsatellite markers. Genetic diversity and forensic parameters were calculated for the overall population, and according to ethno-linguistics and geography. The overall combined power of exclusion (CPE) was 0.9999965412 and the combined match probability 6,28 × 10-19. CPE was highest for the Khoisan Tuu ethnolinguistic group and the Northeast District at 0.9999582029 and 0.9999922652 respectively. CMP ranged from 6.28 × 10-19 (Khoisan Tuu) to 1,02 × 10-18 (Northwest district). Using pairwise genetic distances (FST), analysis of molecular variance (AMOVA), factorial correspondence analysis (FCA), and the unsupervised Bayesian clustering method found in STRUCTURE and TESS, ethno-linguistics were found to have a greater influence on population structure than geography. FCA showed clustering between Bantu and Khoisan, and within the Bantu. This Bantu sub-structuring was not seen with STRUCTURE and TESS, which detected clustering only between Bantu and Khoisan. The patterns of population structure revealed highlight the need for regional reference databases that include ethno-linguistic and geographic location information. These markers have important potential for bio-anthropological studies as well as for forensic applications.


Perseveration moderates the relationship between perfectionism and binge eating: a multi-method daily diary study.
Theory and evidence suggest perfectionism is associated with binge eating. Few studies test conditions under which this association is particularly strong. To better understand the perfectionism-binge eating connection, the present study introduced perseveration as a moderator. A sample of 317 undergraduates completed a computerized Stroop task, baseline self-report measures of perfectionism (i.e., doubts about actions) and binge eating, and self-report daily diary measures of binge eating. Perseveration was defined in terms of reaction time difficulties when consecutive trials required a change of response relative to consecutive trials not requiring a change. Results indicated high levels of doubts about actions were especially related to high levels of binge eating for participants high (versus low) in perseveration. Findings suggest perfectionistic, nagging self-doubts, combined with a tendency to get stuck on thoughts or behaviors, may contribute to increased binge eating-a habitual behavior linked with difficulties in self-regulation.


Public antibodies to malaria antigens generated by two LAIR1 insertion modalities.
In two previously described donors, the extracellular domain of LAIR1, a collagen-binding inhibitory receptor encoded on chromosome 19 (ref. 1), was inserted between the V and DJ segments of an antibody. This insertion generated, through somatic mutations, broadly reactive antibodies against RIFINs, a type of variant antigen expressed on the surface of Plasmodium falciparum-infected erythrocytes. To investigate how frequently such antibodies are produced in response to malaria infection, we screened plasma from two large cohorts of individuals living in malaria-endemic regions. Here we report that 5-10% of malaria-exposed individuals, but none of the European blood donors tested, have high levels of LAIR1-containing antibodies that dominate the response to infected erythrocytes without conferring enhanced protection against febrile malaria. By analysing the antibody-producing B cell clones at the protein, cDNA and gDNA levels, we characterized additional LAIR1 insertions between the V and DJ segments and discovered a second insertion modality whereby the LAIR1 exon encoding the extracellular domain and flanking intronic sequences are inserted into the switch region. By exon shuffling, this mechanism leads to the production of bispecific antibodies in which the LAIR1 domain is precisely positioned at the elbow between the VH and CH1 domains. Additionally, in one donor the genomic DNA encoding the VH and CH1 domains was deleted, leading to the production of a camel-like LAIR1-containing antibody. Sequencing of the switch regions of memory B cells from European blood donors revealed frequent templated inserts originating from transcribed genes that, in rare cases, comprised exons with orientations and frames compatible with expression. These results reveal different modalities of LAIR1 insertion that lead to public and dominant antibodies against infected erythrocytes and suggest that insertion of templated DNA represents an additional mechanism of antibody diversification that can be selected in the immune response against pathogens and exploited for B cell engineering.


[Evaluation of biological and clinical potential of paleolithic diet].
Accumulating evidences suggest that foods that were regularly consumed during the human primates and evolution, in particular during the Paleolithic era (2.6-0.01 x 10(6) years ago), may be optimal for the prevention and treatment of some chronic diseases. It has been postulated that fundamental changes in the diet and other lifestyle conditions that occurred after the Neolithic Revolution, and more recently with the beginning of the Industrial Revolution are too recent taking into account the evolutionary time scale for the human genome to have completely adjust. In contemporary Western populations at least 70% of daily energy intake is provided by foods that were rarely or never consumed by Paleolithic hunter-gatherers, including grains, dairy products as well as refined sugars and highly processed fats. Additionally, compared with Western diets, Paleolithic diets, based on recently published estimates of macronutrient and fatty acid intakes from an East African Paleolithic diet, contained more proteins and long-chain polyunsaturated fatty acids, and less linoleic acid. Observational studies of hunter-gatherers and other non-western populations lend support to the notion that a Paleolithic type diet may reduce the risk of cardiovascular disease, metabolic syndrome, type 2 diabetes, cancer, acne vulgaris and myopia. Moreover, preliminary intervention studies using contemporary diet based on Paleolithic food groups (meat, fish, shellfish, fresh fruits and vegetables, roots, tubers, eggs, and nuts), revealed promising results including favorable changes in risk factors, such as weight, waist circumference, C-reactive protein, glycated haemoglobin (HbAlc), blood pressure, glucose tolerance, insulin secretion, insulin sensitivity and lipid profiles. Low calcium intake, which is often considered as a potential disadvantage of the Paleolithic diet model, should be weighed against the low content of phytates and the low content of sodium chloride, as well as the high amount of net base yielding vegetables and fruits. Increasing number of evidences supports the view that intake of high glycemic foods and insulinotropic dairy products is involved in the pathogenesis and progression of acne vulgaris in Western countries. In this context, diets that mimic the nutritional characteristics of diets found in hunter-gatherers and other non-western populations may have therapeutic value in treating acne vulgaris. Additionally, more studies is needed to determine the impact of gliadin, specific lectins and saponins on intestinal permeability and the pathogenesis of autoimmune diseases.


[Contamination of oyster sea farm with the Norwalk virus: mechanisms and control].
The Norwalk virus(NV) is widely known as a cause of nonbacterial food poisoning, infant diarrhea, and acute gastroenteritis in the winter months between November and March. While it is strongly suspected that NV that is excreted by humans flows into coastal seawaters via rivers and wastewater treatment facilities to contaminate oysters that are grown in farms in the area, light has yet to be shed on the behavior of this virus in the natural environment. We therefore conducted a polymerase chain reaction (PCR) survey of NV levels in the aquatic environment of the oyster bed area of the Shima region in Mie Prefecture, whereupon the NV was detected in marine sediment, oysters, and mule clams even during the summer months, when food poisoning is infrequent. In order to assess their similarity to human-derived strains, the detected viruses and their human-derived counterparts were subjected to genetic analysis, whereupon some of the detected viruses were found to be remarkably similar to those that were previously detected in humans infected with NV. In the interests of examining methods for decontaminating NV-contaminated oysters, we also conducted an assessment on a system of virus decontamination that focuses on seawater temperature and oyster metabolism, using Poliovirus Sabin strain. The decontamination system mentioned above was a closed loop, water circulating system, built on the same principles as those actually in use at oyster farms. Our experiment indicated that at seawater temperatures of both 10 degrees C and 20 degrees C, virus placed into the water tank was rapidly incorporated into the midgut glands of the oysters. Thereafter, when seawater irradiated with UV was circulated, the virus count in the oysters fell from 1/1,000 to 1/10,000 within 6 hours. These results indicated the utility of this system for virus decontamination, suggesting the possibility of significantly alleviating the risk of NV infection in humans by using this system to maintain the seawater temperature within the decontamination tank above a certain temperature, and to perform decontamination with an adequate water flow.


Genetic and environmental transmission of body mass index fluctuation.
This study sought to determine the relationship between body mass index (BMI) fluctuation and cardiovascular disease phenotypes, diabetes, and depression and the role of genetic and environmental factors in individual differences in BMI fluctuation using the extended twin-family model (ETFM). This study included 14,763 twins and their relatives. Health and Lifestyle Questionnaires were obtained from 28,492 individuals from the Virginia 30,000 dataset including twins, parents, siblings, spouses, and children of twins. Self-report cardiovascular disease, diabetes, and depression data were available. From self-reported height and weight, BMI fluctuation was calculated as the difference between highest and lowest BMI after age 18, for individuals 18-80 years. Logistic regression analyses were used to determine the relationship between BMI fluctuation and disease status. The ETFM was used to estimate the significance and contribution of genetic and environmental factors, cultural transmission, and assortative mating components to BMI fluctuation, while controlling for age. We tested sex differences in additive and dominant genetic effects, parental, non-parental, twin, and unique environmental effects. BMI fluctuation was highly associated with disease status, independent of BMI. Genetic effects accounted for ~34 % of variance in BMI fluctuation in males and ~43 % of variance in females. The majority of the variance was accounted for by environmental factors, about a third of which were shared among twins. Assortative mating, and cultural transmission accounted for only a small proportion of variance in this phenotype. Since there are substantial health risks associated with BMI fluctuation and environmental components of BMI fluctuation account for over 60 % of variance in males and over 50 % of variance in females, environmental risk factors may be appropriate targets to reduce BMI fluctuation.


Quantitative 3-dimensional echocardiography for accurate and rapid cardiac phenotype characterization in mice.
Insufficient techniques exist for rapid and reliable phenotype characterization of genetically manipulated mouse models of cardiac dysfunction. We developed a new, robust, 3-dimensional echocardiography (3D-echo) technique and hypothesized that this 3D-echo technique is as accurate as magnetic resonance imaging (MRI) and histology for assessment of left ventricular (LV) volume, ejection fraction, mass, and infarct size in normal and chronically infarcted mice. Using a high-frequency, 7/15-MHz, linear-array ultrasound transducer, we acquired ECG and respiratory-gated, 500-microm consecutive short-axis slices of the murine heart within 4 minutes. The short-axis movies were reassembled off-line in a 3D matrix by using the measured platform locations to position each slice in 3D. Epicardial and endocardial heart contours were manually traced, and a B-spline surface was fitted to the delineated image curves to reconstruct the heart volumes. Excellent correlations were obtained between 3D-echo and MRI for LV end-systolic volumes (r=0.99, P<0.0001), LV end-diastolic volumes (r=0.99, P<0.0001), ejection fraction (r=0.99, P<0.0001), LV mass (r=0.94, P<0.0019), and infarct size (r=0.98, P<0.0001). Also, excellent correlations were found between the 3D-echo-derived LV mass and necropsy LV mass in normal mice (r=0.99, P<0.0001), as well as for 3D-echo-derived infarct size and histologically determined infarct size (r=0.99, P<0.0001) in mice with chronic heart failure. Bland-Altman analysis showed excellent limits of agreement between techniques for all measured parameters. This new, fast, and highly reproducible 3D-echo technique should be of widespread applicability for high-throughput murine cardiac phenotyping studies.


Detecting Cataract Using Smartphones.
Cataract, which is the clouding of the crystalline lens, is the most prevalent eye disease accounting for 51% of all eye diseases in the U.S. Cataract is a progressive disease, and its early detection is critical for preventing blindness. In this paper, an efficient approach to identify cataract disease by adopting luminance features using a smartphone is proposed. Initially, eye images captured by a smartphone were cropped to extract the lens, and the images were preprocessed to remove irrelevant background and noise by utilizing median filter and watershed transformation. Then, a novel luminance transformation from pixel brightness algorithm was introduced to extract lens image features. The luminance and texture features of different types of cataract disease images could be obtained accurately in this stage. Finally, by adopting support vector machines (SVM) as the classification method, cataract eyes were identified. From all the images that we fed into our system, our method could diagnose diseased eyes with 96.6% accuracy, 93.4% specificity, and 93.75% sensitivity. The proposed method provides an affordable, rapid, easy-to-use, and versatile method for detecting cataracts by using smartphones without the use of bulky and expensive imaging devices. This methodcan be used for bedside telemedicine applications or in remote areas that have medical shortages. Previous smartphone-based cataract detection methods include texture feature analysis with 95 % accuracy, Gray Level Co-occurrence Matrix (GLCM) method with 89% accuracy, red reflex measurement method, and RGB color feature extraction method using cascade classifier with 90% accuracy. The accuracy of cataract detection in these studies is subject to changes in smartphone models and/or environmental conditions. However, our novel luminance-based method copes with different smartphone camera sensors and chroma variations, while operating independently from sensors' color characteristics and changes in distances and camera angle. Clinical and Translational Impact-This study is an early/pre-clinical research proposing a novel luminance-based method of detecting cataract using smartphones for remote/at-home monitoring and telemedicine application.


Gastroprotective effects of flavonoids in plant extracts.
The purpose of this paper is to overview the relations between plant-originated substances and their bioactivity measured in terms of antioxidant, cytoprotective and antiulcer activities. In addition, we assessed whether these compounds are capable of affecting the gastric mucosal lesions induced by absolute ethanol applied intragastrically (i.g.). The following plant-originated flavonoid substances were considered; Solon (Sophoradin extract), Amaranth seed extract, grapefruit-seed extract (GSE) and capsaicin (extract of chilly pepper). The area of gastric mucosa lesions and gastric blood flow were measured in rats with ethanol-induced lesions without (control) and with one of the tested substances without and with capsaicin denervation of afferent nerves or administration of L-nitro-arginine (L-NNA), an inhibitor of nitric oxide synthase (NOS). Male Wistar rats, weighing 180-220 g fasted for 24 h before the study where used 100% ethanol was applied i.g. to induce gastric lesions, whose area was determined by planimetry. Gastric blood flow was assessed using electrolytic regional blood flowmeter. All tested plant-originated substances afforded gastroprotection against ethanol-induced damage and this was accompanied by increase in gastric microcirculation, both changes being reversed by pretreatment with neurotoxic dose of capsaicin or by pretreatment with L-NNA. We conclude that plant-originated flavonoid substances are highly gastroprotective probably due to enhancement of the expression of constitutive NOS and release of NO and neuropeptides such as calcitonin gene related peptide (CGRP) released from sensory afferent nerves increasing gastric microcirculation.


Unveiling pathways used by Leishmania amazonensis amastigotes to subvert macrophage function.
This article provides a summary and discussion of properties of Leishmania amazonensis-loaded mouse macrophages. It illustrates how high-throughput analysis is expected to contribute to deciphering features displayed by macrophages when they are subverted as host cells for replicating Leishmania amastigotes. Firstly, we discuss features of mouse mononuclear phagocytes in steady-state conditions, including the phagocytosis of apoptotic cells. Secondly, we discuss results from ongoing investigations aimed at characterizing transcriptional signatures displayed by BALB/c mouse bone marrow-derived macrophages housing replicating L. amazonensis amastigotes. After a brief presentation on the feasibility of high-throughput microscopy relying on our robust culture system, we share some perspectives on the perpetuation of L. amazonensis in their hosts. Within this latter context, a novel question is formulated and its relevance is discussed: do the Leishmania amastigotes that persist within the mammalian dermis reach a non-replicating developmental stage? If so, is this developmental stage the only one displaying the features required for further development as promastigotes within the sand fly gut lumen?


High Incidence of Epileptiform Potentials During Continuous EEG Monitoring in Critically Ill COVID-19 Patients.
Objective: To analyze continuous 1- or 2-channel electroencephalograms (EEGs) of mechanically ventilated patients with coronavirus disease 2019 (COVID-19) with regard to occurrence of epileptiform potentials. Design: Single-center retrospective analysis. Setting: Intensive care unit of Hannover Medical School, Hannover, Germany. Patients: Critically ill COVID-19 patients who underwent continuous routine EEG monitoring (EEG monitor: Narcotrend-Compact M) during sedation. Measurements and Main Results: Data from 15 COVID-19 patients (11 men, four women; age: 19-75 years) were evaluated. Epileptiform potentials occurred in 10 of 15 patients (66.7%). Conclusions: The results of the evaluation regarding the occurrence of epileptiform potentials show that there is an unusually high percentage of cerebral involvement in patients with severe COVID-19. EEG monitoring can be used in COVID-19 patients to detect epileptiform potentials.


Incarceration in fragile families.
Since the mid-1970s the U.S. imprisonment rate has increased roughly fivefold. As Christopher Wildeman and Bruce Western explain, the effects of this sea change in the imprisonment rate--commonly called mass imprisonment or the prison boom--have been concentrated among those most likely to form fragile families: poor and minority men with little schooling. Imprisonment diminishes the earnings of adult men, compromises their health, reduces familial resources, and contributes to family breakup. It also adds to the deficits of poor children, thus ensuring that the effects of imprisonment on inequality are transferred intergenerationally. Perversely, incarceration has its most corrosive effects on families whose fathers were involved in neither domestic violence nor violent crime before being imprisoned. Because having a parent go to prison is now so common for poor, minority children and so negatively affects them, the authors argue that mass imprisonment may increase future racial and class inequality--and may even lead to more crime in the long-term, thereby undoing any benefits of the prison boom. U.S. crime policy has thus, in the name of public safety, produced more vulnerable families and reduced the life chances of their children. Wildeman and Western advocate several policy reforms, such as limiting prison time for drug offenders and for parolees who violate the technical conditions of their parole, reconsidering sentence enhancements for repeat offenders, and expanding supports for prisoners and ex-prisoners. But Wildeman and Western argue that criminal justice reform alone will not solve the problems of school failure, joblessness, untreated addiction, and mental illness that pave the way to prison. In fact, focusing solely on criminal justice reforms would repeat the mistakes the nation made during the prison boom: trying to solve deep social problems with criminal justice policies. Addressing those broad problems, they say, requires a greater social commitment to education, public health, and the employment opportunities of low-skilled men and women. The primary sources of order and stability--public safety in its wide sense--are the informal social controls of family and work. Thus, broad social policies hold the promise not only of improving the wellbeing of fragile families, but also, by strengthening families and providing jobs, of contributing to public safety.


PHASES Score for the Management of Intracranial Aneurysm: A Cross-Sectional Population-Based Retrospective Study.
The aim of this study is to assess whether the PHASES score allows to (1) match decisions taken by multidisciplinary team whether to observe or intervene, (2) classify patients being diagnosed with a ruptured versus unruptured intracranial aneurysm (UIA), and (3) discriminate patients at low risk of rupture from the population of patients diagnosed with intracranial aneurysm. Population-based prospective and consecutive data were collected between 2006 and 2014. Patients (n=841) were stratified into 4 groups: stable UIA; growing observed UIA; immediately treated UIA; and aneurysmal subarachnoid hemorrhage (aSAH). All patients initially observed were pooled in a follow-up UIA group; patients from growing observed UIA, immediately treated UIA, and aSAH were pooled in a high risk of rupture group. Results are expressed as median [quartile 1, quartile 3]. PHASES scores of immediately treated UIA patients were significantly higher than follow-up UIA group (5 [3, 7] versus 2 [1, 4]). Patients diagnosed with UIA and PHASES score of >3 were more likely to be treated, and the score ≤3 was predictive for observation (areas under these curves=0.74). Odds of being diagnosed with an aSAH were associated with PHASES score of >3 (UIA, 4 [2, 6]; aSAH, 5 [4, 8]; areas under these curves=0.66). Scores of stable UIA patients were significantly lower than high risk of rupture group (2 [1, 4] versus 5 [4, 7]; stable UIA outcome prediction by PHASES score of ≤3: areas under these curves=0.76). There is a progression of PHASES score between stable UIA, growing observed UIA, immediately treated UIA, and aSAH groups. PHASES score of ≤3 is associated with a low but not negligible likelihood of aneurysm rupture, and specificity of the classifier is low.


Single-angle-of-incidence ellipsometry.
We introduce single angle-of-incidence (SAI) ellipsometry [U.S. patent application 20070024850 (14 July 2006)] as a technique to completely identify, i.e., totally characterize, film-substrate systems. We show that only one measurement of the ellipsometric function rho at one angle of incidence and one wavelength is totally sufficient to determine the optical constant of the film N1, its thickness d, and the substrate's optical constant N2. Obviously, it is also sufficient for characterizing only the film, determining N1 and d, and for characterizing only the substrate, determining N2 and d, as well as for characterizing only bare substrates. An inverse genetic algorithm (IGA) for complete identification is presented that is based on a physical condition of the transparent-film-absorbing-substrate system . This IGA is used to identify the film-substrate system in four separate cases. We show that removing the film thickness from the fitness function of the genetic algorithm and using the defined optimum population size to characterize the film reduces the computational effort from 20,000 to 69 fitness-function calculations; the number of calculations to characterize an absorbing layer is reduced from 80,000 to 180. This is a very significant reduction and is very welcome in real-time applications. An error analysis is presented that shows that the IGA is resilient to, not affected by, random experimental errors and that it gives very good results in the presence of both random and systematic errors of the ellipsometer system. Experimental results are given that also prove the robustness, stability, and high accuracy of the method. We present data only for the SiO2-Si film-substrate system, but the IGA works for any film-substrate system, physical or not.


Use of a Model Maturity Matrix to Build a Quality Improvement Infrastructure for Psychiatric Care.
Despite a call to action by accreditation bodies and payers alike, the infrastructure needed to continuously improve the quality and safety of behavioral health care has been slow to develop. The causes of this are complex and multifactorial yet likely include underdeveloped leadership and capacity to sustainably improve the quality of psychiatric care. This article proposes the use of a quality improvement maturity matrix to serve as both an implementation strategy and a self-assessment measure to strategically guide this infrastructure development. The matrix describes six phases of development across three domains that are generalizable to various behavioral health care settings.


Ruminants Contribute Fecal Contamination to the Urban Household Environment in Dhaka, Bangladesh.
In Dhaka, Bangladesh, the sensitivity and specificity of three human, three ruminant, and one avian source-associated QPCR microbial source tracking assays were evaluated using fecal samples collected on site. Ruminant-associated assays performed well, whereas the avian and human assays exhibited unacceptable cross-reactions with feces from other hosts. Subsequently, child hand rinses (n = 44) and floor sponge samples (n = 44) from low-income-households in Dhaka were assayed for fecal indicator bacteria (enterococci, Bacteroidales, and Escherichia coli) and a ruminant-associated bacterial target (BacR). Mean enterococci concentrations were of 100 most probable number (MPN)/2 hands and 1000 MPN/225 cm(2) floor. Mean concentrations of Bacteroidales were 10(6) copies/2 hands and 10(5) copies/225 cm(2) floor. E. coli were detected in a quarter of hand rinse and floor samples. BacR was detected in 18% of hand rinse and 27% of floor samples. Results suggest that effective household fecal management should account not only for human sources of contamination but also for animal sources. The poor performance of the human-associated assays in the study area calls into the question the feasibility of developing a human-associated marker in urban slum environments, where domestic animals are exposed to human feces that have been disposed in pits and open drains.


Adoptive transfer of diabetes into immunodeficient NOD-scid/scid mice. Relative contributions of CD4+ and CD8+ T-cells from diabetic versus prediabetic NOD.NON-Thy-1a donors.
Precise definition of the role of both CD4 and CD8 T-cell subsets from NOD mice in the adoptive transfer of diabetes has been complicated by the possibility that endogenous T-cells may be recruited. Two newly created NOD congenic stocks, NOD.NON-Thy-1a and NOD/LtSz-scid, have been used as T-cell donors and recipients, respectively, to eliminate contributions from endogenous T-cells and thus to define the requirement for transferred T-cell subsets as a function of underlying diabetes development in the NOD donor. Total T-cells and T-cell subsets prepared from either prediabetic or diabetic NOD.NON-Thy-1a donors were adoptively transferred into 6-wk-old NOD-scid/scid recipients that were monitored for diabetes development. Both flow cytometric and histological analysis of recipient spleen and pancreas after adoptive transfer showed lymphocytes of donor (Thy1.1+) origin exclusively. Total T-cell and enriched CD4+ T-cell preparations from both diabetic and young prediabetic donors transferred diabetes to NOD-scid/scid recipients. However, the mean time to diabetes onset was doubled when CD4+ lymphocytes were isolated from prediabetic versus diabetic donors, and these transfers were complicated by the generation of small but significant numbers of CD8+ cells over time. Enriched CD8+ populations alone were unable to transfer disease. More rigorous exclusion of CD8+ cells by means of anti-CD8 MoAb treatment in vivo of the recipients of enriched CD4+ cells demonstrated a significant difference in the diabetogenic potency of CD4+ lymphocytes from diabetic versus nondiabetic donors. Diabetes was adoptively transferred to 58% of the recipients of enriched CD4+ lymphocytes from diabetic donors. In contrast, none of the recipients of enriched CD4+ lymphocytes from young prediabetic donors developed diabetes after MoAb treatment in vivo. The ability of a T-cell population to produce severe insulitis and sialitis in NOD-scid/scid recipients of T-cells closely paralleled its ability to induce diabetes. In an effort to suppress insulitis by suppression of macrophage migration to the islets, NOD-scid/scid mice were treated with silica in conjunction with adoptive transfer of T-cells from diabetic donors. Chronic silica treatment failed to deplete tissue macrophages and did not prevent diabetes development after transfer of unfractionated T-cells. Evidence is discussed indicating that the age-associated differences in ability of CD4+ T-cells to adoptively transfer diabetes in the absence of the CD8+ T-cells subset is a function of prior, chronic exposure of the CD4+ lymphocytes to beta-cell antigens in the donor.(ABSTRACT TRUNCATED AT 400 WORDS)


H2B ubiquitylation is part of chromatin architecture that marks exon-intron structure in budding yeast.
The packaging of DNA into chromatin regulates transcription from initiation through 3' end processing. One aspect of transcription in which chromatin plays a poorly understood role is the co-transcriptional splicing of pre-mRNA. Here we provide evidence that H2B monoubiquitylation (H2BK123ub1) marks introns in Saccharomyces cerevisiae. A genome-wide map of H2BK123ub1 in this organism reveals that this modification is enriched in coding regions and that its levels peak at the transcribed regions of two characteristic subgroups of genes. First, long genes are more likely to have higher levels of H2BK123ub1, correlating with the postulated role of this modification in preventing cryptic transcription initiation in ORFs. Second, genes that are highly transcribed also have high levels of H2BK123ub1, including the ribosomal protein genes, which comprise the majority of intron-containing genes in yeast. H2BK123ub1 is also a feature of introns in the yeast genome, and the disruption of this modification alters the intragenic distribution of H3 trimethylation on lysine 36 (H3K36me3), which functionally correlates with alternative RNA splicing in humans. In addition, the deletion of genes encoding the U2 snRNP subunits, Lea1 or Msl1, in combination with an htb-K123R mutation, leads to synthetic lethality. These data suggest that H2BK123ub1 facilitates cross talk between chromatin and pre-mRNA splicing by modulating the distribution of intronic and exonic histone modifications.


A linear amino acid sequence involved in the interaction of t-PA with its endothelial cell receptor.
Endothelial cell receptors for tissue plasminogen activator (t-PA) have been demonstrated recently, and we have sought to identify a region of the t-PA molecule involved in its interaction with these receptors on human umbilical vein endothelial cells. Of three monoclonal antibodies against various regions of t-PA, one directed against the finger region inhibited 125I-t-PA binding to the cells. Synthetic peptides corresponding in amino acid sequences to segments from within the finger region were constructed, and one of these inhibited t-PA binding. This peptide corresponded to residues 7 through 17 of t-PA. The inhibition by this peptide was specific as other peptides from the finger region were inactive. The inhibitory peptide also did not affect the binding of another fibrinolytic ligand, urokinase, to the cells. Although a role for other regions of t-PA in binding to endothelial cells cannot be excluded, the results implicate a short span of linear amino acid sequence within the finger region in the interaction of t-PA with endothelial cells.


Integrating data from multiple Finnish biobanks and national health-care registers for retrospective studies: Practical experiences.
Aim: This case study aimed to investigate the process of integrating resources of multiple biobanks and health-care registers, especially addressing data permit application, time schedules, co-operation of stakeholders, data exchange and data quality. Methods: We investigated the process in the context of a retrospective study: Pharmacogenomics of antithrombotic drugs (PreMed study). The study involved linking the genotype data of three Finnish biobanks (Auria Biobank, Helsinki Biobank and THL Biobank) with register data on medicine dispensations, health-care encounters and laboratory results. Results: We managed to collect a cohort of 7005 genotyped individuals, thereby achieving the statistical power requirements of the study. The data collection process took 16 months, exceeding our original estimate by seven months. The main delays were caused by the congested data permit approval service to access national register data on health-care encounters. Comparison of hospital data lakes and national registers revealed differences, especially concerning medication data. Genetic variant frequencies were in line with earlier data reported for the European population. The yearly number of international normalised ratio (INR) tests showed stable behaviour over time. Conclusions: A large cohort, consisting of versatile individual-level phenotype and genotype data, can be constructed by integrating data from several biobanks and health data registers in Finland. Co-operation with biobanks is straightforward. However, long time periods need to be reserved when biobank resources are linked with national register data. There is a need for efforts to define general, harmonised co-operation practices and data exchange methods for enabling efficient collection of data from multiple sources.


The Expression of Tristetraprolin and Its Relationship with Urinary Proteins in Patients with Diabetic Nephropathy.
Tristetraprolin (TTP), also known as zinc finger protein 36, is an RNA binding protein that has a significant role in regulating the expression of mRNAs containing AU-rich elements. We postulated that TTP might regulate interleukin (IL)-6 and IL-18 expression in diabetes. This study aimed to test the hypothesis that the levels of TTP are correlated with nephropathy in patients with type 2 diabetes. Eighty-seven patients (61.3±9.6 years old) who had been diagnosed with type 2 diabetes mellitus and 41 age and sex matched healthy control subjects were enrolled. The diabetes patients were classified into those without proteinuria, with microalbuminuria, and with clinical proteinuria groups according to the ratio of urinary excretion of albumin/creatinine (ACR). Serum and urinary levels of IL-6 and IL-18 were significantly elevated, but those of TTP were significantly decreased in patients with diabetes as compared with control subjects. In addition, serum and urinary levels of IL-6 and IL-18 were significantly higher, but those of TTP were significantly lower in patients with proteinuria than in patients without proteinuria or with microalbuminuria. There was a significant correlation between serum TTP and IL-6/IL-18 (correlation coefficients of -0.572 and -0.685, P < 0.05). These results show that diabetes with clinical proteinuria is accompanied by decreased urinary and serum level of TTP and increased levels of IL-6 and IL-18. Decreased TTP expression might occur prior to the increase in IL-6 and IL-18, and decrease of TTP might provide an earlier marker for glomerular dysfunction than IL-6 and IL-18.


Reconstructing two-dimensional acoustic object fields by use of digital phase conjugation of scanning laser vibrometry recordings.
A scanning laser Doppler vibrometer is used to record two-dimensional ultrasound fields in air. The laser light of the vibrometer traverses the sound field to and from a rigid reflector and determines the velocity field, a quantity proportional to the sound pressure rate, in each scanned point relative to the sound source. The object sound is the scattered field from objects outside the recording area. Digital reconstruction by use of phase conjugation (time reversal) of the object sound field is then performed, and the original object field intensity and phase is reconstructed.


Pathways between childhood trauma, intimate partner violence, and harsh parenting: findings from the UN Multi-country Study on Men and Violence in Asia and the Pacific.
Although childhood trauma and violence against women are global public health issues, few population-based data from low-income and middle-income countries exist about the links between them. We present data from the UN Multi-country Study on Men and Violence in Asia and the Pacific, exploring the pathways between different forms of childhood trauma and violence against women. In this multicountry study, we interviewed multistage representative samples of men and women, aged 18-49 years, in Asia and the Pacific, using standardised population-based household surveys. Men were interviewed in six countries, and women in four. Respondents were asked questions about their perpetration or experience of intimate partner violence or non-partner sexual violence, childhood trauma, and harsh parenting (smacking their children as a form of discipline). We used maximum likelihood multivariate logit models to explore associations between childhood trauma and violence against women, and fitted path models to explore associations between experience and perpetration of child maltreatment. Between Jan 1, 2011, and Dec 1, 2012, 10 178 men and 3106 women completed interviews in this study, with between 815 and 1812 men per site and 477 and 1103 women per site. The proportion of men who experienced any childhood trauma varied between 59% (n=478, 95% CI 54·0-63·3; Indonesia rural site) and 92% (n=791, 89·4-93·8; Bougainville, Papua New Guinea). For women, the results ranged from 44% (n=272, 37·7-50·8; Sri Lanka) to 84% (n=725, 80·7-86·8; Bougainville, Papua New Guinea). For men, all forms of childhood trauma were associated with all forms of intimate partner violence perpetration. For women, all forms of childhood trauma were associated with physical intimate partner violence, and both physical and sexual intimate partner violence. There were significant, often gendered, pathways between men's and women's perpetration and experiences of childhood trauma, physical intimate partner violence, harsh parenting, and other factors. The data point to both a co-occurrence and a cycle of abuse, with childhood trauma leading to violence against women and further child maltreatment, which in turn increases the risk of experience or perpetration of violence during adulthood. Efforts to prevent both forms of violence would benefit from a meaningful integrated approach. Interventions should promote positive parenting, address inequality and the normalisation of violence across the life course, and transform men's power over women and children. Partners for Prevention. National studies were funded by the UN Population Fund in Bangladesh and China, UN Women in Cambodia and Indonesia, UN Develoment Programme in Papua New Guinea, and CARE in Sri Lanka.


JS-K, a glutathione/glutathione S-transferase-activated nitric oxide releasing prodrug inhibits androgen receptor and WNT-signaling in prostate cancer cells.
Nitric oxide (NO) and its oxidative reaction products have been repeatedly shown to block steroid receptor function via nitrosation of zinc finger structures in the DNA-binding domain (DBD). In consequence NO-donors could be of special interest for the treatment of deregulated androgen receptor(AR)-signaling in castration resistant prostate cancer (CRPC). Prostate cancer (PCa) cells were treated with JS-K, a diazeniumdiolate derivate capable of generating large amounts of intracellular NO following activation by glutathione S-transferase. Generation of NO was determined indirectly by the detection of nitrate in tissue culture medium or by immunodetection of nitrotyrosine in the cytoplasm. Effects of JS-K on intracellular AR-levels were determined by western blotting. AR-dimerization was analyzed by mammalian two hybrid assay, nuclear translocation of the AR was visualized in PCa cells transfected with a green fluorescent AR-Eos fusion protein using fluorescence microscopy. Modulation of AR- and WNT-signalling by JS-K was investigated using reporter gene assays. Tumor cell proliferation following JS-K treatment was measured by MTT-Assay. The NO-releasing compound JS-K was shown to inhibit AR-mediated reporter gene activity in 22Rv1 CRPC cells. Inhibition of AR signaling was neither due to an inhibition of nuclear import nor to a reduction in AR-dimerization. In contrast to previously tested NO-donors, JS-K was able to reduce the intracellular concentration of functional AR. This could be attributed to the generation of extremely high intracellular levels of the free radical NO as demonstrated indirectly by high levels of nitrotyrosine in JS-K treated cells. Moreover, JS-K diminished WNT-signaling in AR-positive 22Rv1 cells. In line with these observations, castration resistant 22Rv1 cells were found to be more susceptible to the growth inhibitory effects of JS-K than the androgen dependent LNCaP which do not exhibit an active WNT-signaling pathway. Our results suggest that small molecules able to inhibit WNT- and AR-signaling via NO-release represent a promising platform for the development of new compounds for the treatment of CRPC.


Common developmental pathways link tooth shape to regeneration.
In many non-mammalian vertebrates, adult dentitions result from cyclical rounds of tooth regeneration wherein simple unicuspid teeth are replaced by more complex forms. Therefore and by contrast to mammalian models, the numerical majority of vertebrate teeth develop shape during the process of replacement. Here, we exploit the dental diversity of Lake Malawi cichlid fishes to ask how vertebrates generally replace their dentition and in turn how this process acts to influence resulting tooth morphologies. First, we used immunohistochemistry to chart organogenesis of continually replacing cichlid teeth and discovered an epithelial down-growth that initiates the replacement cycle via a labial proliferation bias. Next, we identified sets of co-expressed genes from common pathways active during de novo, lifelong tooth replacement and tooth morphogenesis. Of note, we found two distinct epithelial cell populations, expressing markers of dental competence and cell potency, which may be responsible for tooth regeneration. Related gene sets were simultaneously active in putative signaling centers associated with the differentiation of replacement teeth with complex shapes. Finally, we manipulated targeted pathways (BMP, FGF, Hh, Notch, Wnt/β-catenin) in vivo with small molecules and demonstrated dose-dependent effects on both tooth replacement and tooth shape. Our data suggest that the processes of tooth regeneration and tooth shape morphogenesis are integrated via a common set of molecular signals. This linkage has subsequently been lost or decoupled in mammalian dentitions where complex tooth shapes develop in first generation dentitions that lack the capacity for lifelong replacement. Our dissection of the molecular mechanics of vertebrate tooth replacement coupled to complex shape pinpoints aspects of odontogenesis that might be re-evolved in the lab to solve problems in regenerative dentistry.


Just-in-time classifiers for recurrent concepts.
Just-in-time (JIT) classifiers operate in evolving environments by classifying instances and reacting to concept drift. In stationary conditions, a JIT classifier improves its accuracy over time by exploiting additional supervised information coming from the field. In nonstationary conditions, however, the classifier reacts as soon as concept drift is detected; the current classification setup is discarded and a suitable one activated to keep the accuracy high. We present a novel generation of JIT classifiers able to deal with recurrent concept drift by means of a practical formalization of the concept representation and the definition of a set of operators working on such representations. The concept-drift detection activity, which is crucial in promptly reacting to changes exactly when needed, is advanced by considering change-detection tests monitoring both inputs and classes distributions.


Association of variants in HTRA1 and NOTCH3 with MRI-defined extremes of cerebral small vessel disease in older subjects.
We report a composite extreme phenotype design using distribution of white matter hyperintensities and brain infarcts in a population-based cohort of older persons for gene-mapping of cerebral small vessel disease. We demonstrate its application in the 3C-Dijon whole exome sequencing (WES) study (n = 1924, nWESextremes = 512), with both single variant and gene-based association tests. We used other population-based cohort studies participating in the CHARGE consortium for replication, using whole exome sequencing (nWES = 2,868, nWESextremes = 956) and genome-wide genotypes (nGW = 9924, nGWextremes = 3308). We restricted our study to candidate genes known to harbour mutations for Mendelian small vessel disease: NOTCH3, HTRA1, COL4A1, COL4A2 and TREX1. We identified significant associations of a common intronic variant in HTRA1, rs2293871 using single variant association testing (Pdiscovery = 8.21 × 10-5, Preplication = 5.25 × 10-3, Pcombined = 4.72 × 10-5) and of NOTCH3 using gene-based tests (Pdiscovery = 1.61 × 10-2, Preplication = 3.99 × 10-2, Pcombined = 5.31 × 10-3). Follow-up analysis identified significant association of rs2293871 with small vessel ischaemic stroke, and two blood expression quantitative trait loci of HTRA1 in linkage disequilibrium. Additionally, we identified two participants in the 3C-Dijon cohort (0.4%) carrying heterozygote genotypes at known pathogenic variants for familial small vessel disease within NOTCH3 and HTRA1. In conclusion, our proof-of-concept study provides strong evidence that using a novel composite MRI-derived phenotype for extremes of small vessel disease can facilitate the identification of genetic variants underlying small vessel disease, both common variants and those with rare and low frequency. The findings demonstrate shared mechanisms and a continuum between genes underlying Mendelian small vessel disease and those contributing to the common, multifactorial form of the disease.


Details of risk-benefit communication in informed consent documents for phase I/II trials.
Informed consent documents for clinical studies should disclose all reasonably foreseeable risks and benefits. Little guidance exists on how to navigate the complexities of risk-benefit communication, especially in early clinical research. Practice-oriented development of such guidance should be informed by evidence on what and how details of risks and benefits are currently communicated. We surveyed the responsible parties of phase I/II trials registered in ClinicalTrials.gov that started 2007 or later and completed between 2012 and 2016 to sample informed consent documents from a broad spectrum of early phase clinical trials. Based on an assessment matrix, we qualitatively and quantitatively assessed the informed consent documents for details of risk-benefit communication. The risk-benefit communication in the 172 informed consent documents differed substantially in several regards. The outcome, extent, and likelihood of health risks, for example, were described in 83%, 32%, and 63% of the informed consent documents. Only 45% of informed consent documents specified the outcome of mentioned health benefits, and the extent and likelihood of health benefits were never specified. From those informed consent documents reporting risk likelihoods, only 57% added frequency numbers to words such as "common" or "rare," and even in these cases, we found strong variations for presented frequency ranges. Substantial heterogeneity also exists for how informed consent documents communicate other risk and benefit types and related safeguards. Our study points to several shortcomings and heterogeneities in how informed consent documents communicate risks and benefits to potential research participants. Health risks, for example, should be specified with frequency numbers, and health benefits should be specified at least by mentioning their outcomes. Further demand for research and policy development is needed to harmonize risk-benefit communication and to clarify ways to specify the likelihood of health benefits.


High-glucose environment enhanced oxidative stress and increased interleukin-8 secretion from keratinocytes: new insights into impaired diabetic wound healing.
Impaired wound healing frequently occurs in patients with diabetes. Interleukin (IL)-8 production by keratinocyte is responsible for recruiting neutrophils during healing. Intense inflammation is associated with diabetic wounds, while reduction of neutrophil infiltration is associated with enhanced healing. We hypothesized that increased neutrophil recruitment by keratinocytes may contribute to the delayed healing of diabetic wounds. Using cultured human keratinocytes and a diabetic rat model, the current study shows that a high-glucose environment enhanced IL-8 production via epidermal growth factor receptor (EGFR)-extracellular signal-regulated kinase (ERK) pathway in a reactive oxygen species (ROS)-dependent manner in keratinocytes. In addition, diabetic rat skin showed enhanced EGFR, ERK, and IL-8 expression compared with control rats. The dermal neutrophil infiltration of the wound, as represented by expression of myeloperoxidase level, was also significantly higher in diabetic rats. Treating diabetic rats with dapsone, an agent known to inhibit neutrophil function, was associated with improved healing. In conclusion, IL-8 production and neutrophil infiltration are increased in a high-glucose environment due to elevated ROS level and contributed to impaired wound healing in diabetic skin. Targeting these dysfunctions may present novel therapeutic approaches.


The EUROASPIRE surveys: lessons learned in cardiovascular disease prevention.
The Joint European Societies (JES) guidelines on cardiovascular disease (CVD) prevention published in 1994, 1998, 2003, 2007, 2012 and 2016 defined lifestyle and risk factors targets for patients with coronary or other atherosclerotic disease and people at high risk of developing CVD. Guideline implementation in Europe has been evaluated with four cross-sectional EUROASPIRE surveys starting in mid-1990s. The results showed poor lifestyle and risk factor management in patients with CHD and in high CVD risk people with high prevalences of smoking, obesity, central obesity and diabetes. The control of blood pressure, lipids and glucose was far from optimal. A comparison across the recent three surveys provided a unique description of time trends for secondary prevention over a period of 14 years. The results showed adverse lifestyle trends, a substantial increase in obesity, central obesity and diabetes. Despite significant improvement of blood pressure and lipid control many patients were not reaching the risk factor goals and there was no change in glucose control. Comparing the most recent two surveys, there were no major differences in lifestyle and risk factor management in people at high risk of developing CVD. A new approach to CVD prevention integrating primary and secondary prevention into a modern preventive cardiology programme, focusing on lifestyle and risk factor management is required, in order to reduce the risk of CVD events and improve quality of life in patients with CHD and those at high risk of developing CVD.


Functional similarity analysis of human virus-encoded miRNAs.
miRNAs are a class of small RNAs that regulate gene expression via RNA silencing machinery. Some viruses also encode miRNAs, contributing to the complex virus-host interactions. A better understanding of viral miRNA functions would be useful in designing new preventive strategies for treating diseases induced by viruses. To meet the challenge for how viruses module host gene expression by their encoded miRNAs, we measured the functional similarities among human viral miRNAs by using a method we reported previously. Higher order functions regulated by viral miRNAs were also identified by KEGG pathway analysis on their targets. Our study demonstrated the biological processes involved in virus-host interactions via viral miRNAs. Phylogenetic analysis suggested that viral miRNAs have distinct evolution rates compared with their corresponding genome.


IRE1α-XBP1s pathway promotes prostate cancer by activating c-MYC signaling.
Activation of endoplasmic reticulum (ER) stress/the unfolded protein response (UPR) has been linked to cancer, but the molecular mechanisms are poorly understood and there is a paucity of reagents to translate this for cancer therapy. Here, we report that an IRE1α RNase-specific inhibitor, MKC8866, strongly inhibits prostate cancer (PCa) tumor growth as monotherapy in multiple preclinical models in mice and shows synergistic antitumor effects with current PCa drugs. Interestingly, global transcriptomic analysis reveal that IRE1α-XBP1s pathway activity is required for c-MYC signaling, one of the most highly activated oncogenic pathways in PCa. XBP1s is necessary for optimal c-MYC mRNA and protein expression, establishing, for the first time, a direct link between UPR and oncogene activation. In addition, an XBP1-specific gene expression signature is strongly associated with PCa prognosis. Our data establish IRE1α-XBP1s signaling as a central pathway in PCa and indicate that its targeting may offer novel treatment strategies.


2'-O-alkyloligoribonucleotides, synthesis and applications in molecular biology.
Oligo(2'-O-alkylribonucleotides) have been synthesized in which alkyl is methyl, allyl and butyl. The various phosphoramidite monomers of 2'-O-alkyl uridine, cytidine, adenosine, guanosine, inosine and 2,6-diaminopurine riboside have been synthesized from a minimum of key intermediates. Extra protection of the lactam function in uracil and hypoxanthine proves useful. The high stability of the oligomers combined with incorporation of non-radioactive reporter groups such as fluorophores, biotin and 2,4-dinitrophenylamino (DNP) moities renders them as excellent antisense tools for studying RNA processing, for locating and visualising RNA and RNP complexes in cells, for examining splicing complexes by electron microscopy and for the affinity chromatography of RNA or RNP complexes.


Experiences by patients with asthma-like symptoms of a problem-based learning health education programme.
Eighteen patients with asthma-like symptoms but negative asthma tests, randomly selected, participated in a problem-based learning health education programme. The programme had a multidisciplinary approach and included exercises inspired by cognitive behavioural therapy. The aim of this study was to describe how the patients experienced the programme. After termination of the programme, semi-structured interviews with a phenomenographic approach were conducted with 15 of the participants. Two categories emerged, with three sub-categories each. In these, the informants described how they felt solidarity with the group, received confirmation from the other participants and had increased their self-confidence. They had started to look upon themselves and the disorder from a different perspective, they could describe the disorder in words and they had started to use new, conscious coping strategies. Patients with asthma-like symptoms benefit from taking part in a problem-based learning health education programme. It helps them to reflect upon themselves and the disorder and to use new strategies to cope with it.


Hereditary intrinsic factor deficiency in chaldeans.
Juvenile vitamin B(12) or cobalamin (Cbl) deficiency is notoriously difficult to explain due to numerous acquired and inherited causes. The consequences of insufficient Cbl are megaloblastic anemia, nutrient malabsorption, and neurological problems. The treatment is straightforward with parenteral Cbl supplementation that resolves most health issues without an urgent need to clarify their cause. Aside from being clinically unsatisfying, failing to elucidate the basis of Cbl deficiency means important information regarding recurrence risk is not available to the individual if the cause is contagious or inherited. Acquired causes have largely disappeared in the Modern World because they were mostly due to parasites or malnutrition. Today, perhaps the most common causes of juvenile Cbl deficiency are Imerslund-Gräsbeck syndrome and inherited intrinsic factor deficiency (IFD). Three genes are involved and genetic testing is complicated and not widely available. We used self-identified ancestry to accelerate and confirm the genetic diagnosis of IFD in three families of Chaldean origin. A founder mutation limited to Chaldeans from Iraq in the intrinsic factor gene GIF was identified as the cause. World events reshape the genetic structure of populations and inherited diseases in many ways. In this case, all the patients were diagnosed in the USA among recent immigrants from a single region. While IFD itself is not restricted to one kind of people, certain mutations are limited in their range but migrations relocate them along with their host population. As a result, self-identified ancestry as a stratifying characteristic should perhaps be considered in diagnostic strategies for rare genetic disorders.


Progressive disability in elderly population among tribals of Telangana: a cross sectional study.
The tribal population of Telangana, India, lives in remote and difficult conditions. This study was carried out to find out estimate, the prevalence and progression of disability in elderly population among tribals of Khammam District, Telangana state, India. A population based cross sectional survey was conducted in villages of Tribal Sub Plan area. Elderly people who are 60 years or older were chosen with a two stage sampling procedure: (1) probability proportion to size was used to select clusters and (2) in each selected cluster households were selected by systematic random sampling. The participants were interviewed with the 36 item Telugu version of the World Health Organization Disability Assessment Schedule (WHODAS 2.0) questionnaire. Socio- demographic information, behavioral measurements, health and social benefit indicators were also assessed. Descriptive analytical methods were used for prevalence estimation and logistic regression was used to examine the associations of progressive age over disability among elderly. A total of 506 elderly people from 1349 households in 20 villages across 31mandals of Khammam were interviewed. Majority of elderly population among tribals were illiterate (men 88.94%; women 99.33%), used tobacco (men 81.25%; women 57.72%), consumed alcohol (men 80.77%; women 47.32%) and were hypertensive (men 53.85%; women 63.42%). The prevalence of disability was higher in women. Maximum disability in the interviewed elderly population was seen in domains of performing house hold activities, and mobility. In comparison with men, women expressed more disability for majority of domains. As age progressed, the disability for self-care domain increased to a maximum of 2.6 times in men and 6.6 times in women and for mobility domain increased to a maximum of 9.7 times in men and 7.2 times in women. Although present disability modifying mobility Assistive Devices (AD) can help elderly in overcoming disability, these are primarily designed for built environments. As the needs, cultural sensitivities, and living environment of elderly population in tribals are unique, newer innovative assistive devices should be designed and developed.


Pre-cART Elevation of CRP and CD4+ T-Cell Immune Activation Associated With HIV Clinical Progression in a Multinational Case-Cohort Study.
Despite the success of combination antiretroviral therapy (cART), a subset of HIV-infected patients who initiate cART develop early clinical progression to AIDS; therefore, some cART initiators are not fully benefitted by cART. Immune activation pre-cART may predict clinical progression in cART initiators. A case-cohort study (n = 470) within the multinational Prospective Evaluation of Antiretrovirals in Resource-Limited Settings clinical trial (1571 HIV treatment-naive adults who initiated cART; CD4 T-cell count <300 cells/mm; 9 countries) was conducted. A subcohort of 30 participants per country was randomly selected; additional cases were added from the main cohort. Cases [n = 236 (random subcohort 36; main cohort 200)] had clinical progression (incident WHO stage 3/4 event or death) within 96 weeks after cART initiation. Immune activation biomarkers were quantified pre-cART. Associations between biomarkers and clinical progression were examined using weighted multivariable Cox-proportional hazards models. Median age was 35 years, 45% were women, 49% black, 31% Asian, and 9% white. Median CD4 T-cell count was 167 cells per cubic millimeter. In multivariate analysis, highest quartile C-reactive protein concentration [adjusted hazard ratio (aHR), 2.53; 95% confidence interval (CI): 1.02 to 6.28] and CD4 T-cell activation (aHR, 5.18; 95% CI: 1.09 to 24.47) were associated with primary outcomes, compared with lowest quartiles. sCD14 had a trend toward association with clinical failure (aHR, 2.24; 95% CI: 0.96 to 5.21). Measuring C-reactive protein and CD4 T-cell activation may identify patients with CD4 T-cell counts <300 cells per cubic millimeter at risk for early clinical progression when initiating cART. Additional vigilance and symptom-based screening may be required in this subset of patients even after beginning cART.


Evaluation of Antimicrobial Durability and Anti-Biofilm Effects in Urinary Catheters Against Enterococcus faecalis Clinical Isolates and Reference Strains.
Enterococcus faecalis, Escherichia coli, Staphylococcus epidermidis, Pseudomonas aeruginosa and Candida albicans biofilms are major causes of catheter-associated urinary tract infections. Antimicrobial-coated or impregnated urinary catheters are seen as a possible way to prevent these infections. To determine the biofilm-forming ability of 89 E. faecalis isolates from urinary tract infections and to compare several urinary catheters for antimicrobial durability and the inhibitory effects on biofilm formation of different laboratory strains and clinical isolates of E. faecalis. In vitro experimental study. The biofilm forming ability of E. faecalis isolates was determined by the crystal violet staining and plate counting methods. For comparison of urinary catheters, biofilms of 45 E. faecalis isolates from the catheter samples of hospitalized patients and five laboratory strains of E. coli ATCC25922, S. epidermidis ATCC35984, P. aeruginosa ATCC27853, E. faecalis ATCC29212 and C. albicans ATCC90028 were formed on the catheters in 24-well tissue culture plates. Scanning electron microscopy analysis was performed to observe biofilms. All 89 E. faecalis isolates were found to be biofilm positive. Nitrofurazone-impregnated catheters significantly reduced the cell counts of E. faecalis isolates and completely inhibited the formation of P. aeruginosa and S. epidermidis biofilms compared with the others. Regarding reduction of biofilm cell counts, a hydrophilic-coated catheter was more effective against P. aeruginosa, whereas a silver-coated catheter was found to be more effective against S. epidermidis. The nitrofurazone-impregnated catheter had the best antimicrobial durability. Urine isolates of E. faecalis had considerable ability with respect to biofilm formation. The nitrofurazone-impregnated catheter was the most effective against all tested bacteria; however, the effect of a hydrophilic or silver-coated catheter depends on the species present in it.


A novel surveillance approach for disaster mental health.
Disasters have substantial consequences for population mental health. Social media data present an opportunity for mental health surveillance after disasters to help identify areas of mental health needs. We aimed to 1) identify specific basic emotions from Twitter for the greater New York City area during Hurricane Sandy, which made landfall on October 29, 2012, and to 2) detect and map spatial temporal clusters representing excess risk of these emotions. We applied an advanced sentiment analysis on 344,957 Twitter tweets in the study area over eleven days, from October 22 to November 1, 2012, to extract basic emotions, a space-time scan statistic (SaTScan) and a geographic information system (QGIS) to detect and map excess risk of these emotions. Sadness and disgust were among the most prominent emotions identified. Furthermore, we noted 24 spatial clusters of excess risk of basic emotions over time: Four for anger, one for confusion, three for disgust, five for fear, five for sadness, and six for surprise. Of these, anger, confusion, disgust and fear clusters appeared pre disaster, a cluster of surprise was found peri disaster, and a cluster of sadness emerged post disaster. We proposed a novel syndromic surveillance approach for mental health based on social media data that may support conventional approaches by providing useful additional information in the context of disaster. We showed that excess risk of multiple basic emotions could be mapped in space and time as a step towards anticipating acute stress in the population and identifying community mental health need rapidly and efficiently in the aftermath of disaster. More studies are needed to better control for bias, identify associations with reliable and valid instruments measuring mental health, and to explore computational methods for continued model-fitting, causal relationships, and ongoing evaluation. Our study may be a starting point also for more fully elaborated models that can either prospectively detect mental health risk using real-time social media data or detect excess risk of emotional reactions in areas that lack efficient infrastructure during and after disasters. As such, social media data may be used for mental health surveillance after large scale disasters to help identify areas of mental health needs and to guide us in our knowledge where we may most effectively intervene to reduce the mental health consequences of disasters.


Recent Progress in Detection and Profiling of Cancer Cell-Derived Exosomes.
Exosomes, known as nanometer-sized vesicles (30-200 nm), are secreted by many types of cells. Cancer-derived exosomes have great potential to be biomarkers for early clinical diagnosis and evaluation of cancer therapeutic efficacy. Conventional detection methods are limited to low sensitivity and reproducibility. There are hundreds of papers published with different detection methods in recent years to address these challenges. Therefore, in this review, pioneering researches about various detection strategies are comprehensively summarized and the analytical performance of these tests is evaluated. Furthermore, the exosome molecular composition (protein and nucleic acid) profiling, a single exosome profiling, and their application in clinical cancer diagnosis are reviewed. Finally, the principles and applications of machine learning method in exosomes researches are presented.


In-fiber strain characterization of fiber-optic connector assemblies by Bragg grating sensors.
In-fiber Bragg grating sensors were used to study mechanical strain in optical fibers that were terminated in standard-termination and ribbon connectors. Our findings indicate that terminated sensors experience a compressive strain whose magnitude depends on the cure profile of the epoxy encapsulant used in these connectors. Anneal treatments on these connectors generally reduce the mechanical stress by inducing stress relaxation in the encapsulant layer. These experiments demonstrate the viability of in-fiber sensors to characterize fiber-optic connector assemblies during and following termination.


C-H Alkenylation of Heteroarenes: Mechanism, Rate, and Selectivity Changes Enabled by Thioether Ligands.
Thioether ancillary ligands have been identified that can greatly accelerate the C-H alkenylation of O-, S-, and N-heteroarenes. Kinetic data suggest thioether-Pd-catalyzed reactions can be as much as 800× faster than classic ligandless systems. Furthermore, mechanistic studies revealed C-H bond cleavage as the turnover-limiting step, and that rate acceleration upon thioether coordination is correlated to a change from a neutral to a cationic pathway for this key step. The formation of a cationic, low-coordinate catalytic intermediate in these reactions may also account for unusual catalyst-controlled site selectivity wherein C-H alkenylation of five-atom heteroarenes can occur under electronic control with thioether ligands even when this necessarily involves reaction at a more hindered C-H bond. The thioether effect also enables short reaction times under mild conditions for many O-, S-, and N-heteroarenes (55 examples), including examples of late-stage drug derivatization.


Projective ART for clustering data sets in high dimensional spaces.
A new neural network architecture (PART) and the resulting algorithm are proposed to find projected clusters for data sets in high dimensional spaces. The architecture is based on the well known ART developed by Carpenter and Grossberg, and a major modification (selective output signaling) is provided in order to deal with the inherent sparsity in the full space of the data points from many data-mining applications. This selective output signaling mechanism allows the signal generated in a node in the input layer to be transmitted to a node in the clustering layer only when the signal is similar to the top-down weight between the two nodes and, hence, PART focuses on dimensions where information can be found. Illustrative examples are provided, simulations on high dimensional synthetic data and comparisons with Fuzzy ART module and PROCLUS are also reported.


A new algorithm for the evaluation of shotgun peptide sequencing in proteomics: support vector machine classification of peptide MS/MS spectra and SEQUEST scores.
Shotgun tandem mass spectrometry-based peptide sequencing using programs such as SEQUEST allows high-throughput identification of peptides, which in turn allows the identification of corresponding proteins. We have applied a machine learning algorithm, called the support vector machine, to discriminate between correctly and incorrectly identified peptides using SEQUEST output. Each peptide was characterized by SEQUEST-calculated features such as delta Cn and Xcorr, measurements such as precursor ion current and mass, and additional calculated parameters such as the fraction of matched MS/MS peaks. The trained SVM classifier performed significantly better than previous cutoff-based methods at separating positive from negative peptides. Positive and negative peptides were more readily distinguished in training set data acquired on a QTOF, compared to an ion trap mass spectrometer. The use of 13 features, including four new parameters, significantly improved the separation between positive and negative peptides. Use of the support vector machine and these additional parameters resulted in a more accurate interpretation of peptide MS/MS spectra and is an important step toward automated interpretation of peptide tandem mass spectrometry data in proteomics.


A Generalized Methodology for Data Analysis.
Based on a critical analysis of data analytics and its foundations, we propose a functional approach to estimate data ensemble properties, which is based entirely on the empirical observations of discrete data samples and the relative proximity of these points in the data space and hence named empirical data analysis (EDA). The ensemble functions include the nonparametric square centrality (a measure of closeness used in graph theory) and typicality (an empirically derived quantity which resembles probability). A distinctive feature of the proposed new functional approach to data analysis is that it does not assume randomness or determinism of the empirically observed data, nor independence. The typicality is derived from the discrete data directly in contrast to the traditional approach, where a continuous probability density function is assumed a priori. The typicality is expressed in a closed analytical form that can be calculated recursively and, thus, is computationally very efficient. The proposed nonparametric estimators of the ensemble properties of the data can also be interpreted as a discrete form of the information potential (known from the information theoretic learning theory as well as the Parzen windows). Therefore, EDA is very suitable for the current move to a data-rich environment, where the understanding of the underlying phenomena behind the available vast amounts of data is often not clear. We also present an extension of EDA for inference. The areas of applications of the new methodology of the EDA are wide because it concerns the very foundation of data analysis. Preliminary tests show its good performance in comparison to traditional techniques.


Cost-effectiveness analyses of durvalumab consolidation therapy versus no consolidation therapy after chemoradiotherapy in stage-III NSCLC.
Non-small cell lung cancer (NSCLC) is the most common form of lung cancer. Recently, Durvalumab was approved as a potential immunotherapy for the management of unresectable stage III NSCLC. Economic studies from different parts of the world presented varying findings. Therefore, the objective of this study was to assess the cost-effectiveness of durvalumab consolidation therapy versus no consolidation therapy in patients with unresectable stage III NSCLC. PubMed, Embase, and Cochrane databases were searched till March 2022 to identify all the studies assessing the economic evaluation of durvalumab in patients with unresectable stage III NSCLC who had not progressed after chemoradiotherapy. Eligible studies were screened by two reviewers independently and the quality of included studies was evaluated using the updated version of Consolidated Health Economic Evaluation Reporting Standards (CHEERS 2022) checklists. All costs were converted to 2022 US dollars ($) by adjusting for the gross domestic product (GDP) deflator index and purchasing power parities for GDP. A total of seven studies were found to be eligible for inclusion. The majority of studies were conducted in the US (n = 3), while one study each was conducted in China, Italy, Switzerland, and the UK. The healthcare payers' perspective was most commonly observed among the included studies and the time horizon varied from 5 years to a lifetime. Three studies received funding from Industry. Four included studies used the Markov model, while two employed the semi-Markov model and, one study used decision-analytic model. The ICER of durvalumab consolidation therapy in the US was found to be in the range of $59,850 to $145,543 per Quality-Adjusted Life Years (QALY). Likewise, the ICER of durvalumab in European countries ranged from $62,021 to $76,068 per QALY. The ICER was below the implemented country-specific willingness-to-pay thresholds in all the included studies. Durvalumab consolidation therapy was found to be cost-effective compared to no consolidation therapy after chemoradiotherapy in stage-III NSCLC patients.


Proliferative response of gingival cells to ultrasonic and/or vibration toothbrushes.
To evaluate the effects of ultrasonic and/or vibration toothbrushes on cell proliferation and collagen synthesis. In eight dogs, teeth and gingivae were stimulated once a day as follows: the first quadrant with an ultrasonic toothbrush (1.6 MHz); the second one with a mechanical vibratory toothbrush (141 Hz); and the third one with a toothbrush generating both the ultrasound and the vibration. The fourth quadrant served as a control. Proliferative activity and collagen synthesis of gingival cells were evaluated by assaying the expression of proliferating cell nuclear antigen (PCNA) and procollagen type I C-peptide (PIP), respectively. After 5 weeks, ultrasonic or vibratory toothbrushes increased the numbers of PCNA-positive fibroblasts and PIP-positive fibroblasts. Toothbrushing with a combination of ultrasound and mechanical vibration increased the numbers of PIP-positive fibroblasts, total fibroblasts and vascular endothelial cells to a greater extent than the one with only ultrasound alone. Vibratory toothbrush, but not the ultrasonic one, induced an increase in collagen density without gingival overgrowth.


Systemic delivery of recombinant proteins by genetically modified myoblasts.
The ability to stably deliver recombinant proteins to the systemic circulation would facilitate the treatment of a variety of acquired and inherited diseases. To explore the feasibility of the use of genetically engineered myoblasts as a recombinant protein delivery system, stable transfectants of the murine C2C12 myoblast cell line were produced that synthesize and secrete high levels of human growth hormone (hGH) in vitro. Mice injected with hGH-transfected myoblasts had significant levels of hGH in both muscle and serum that were stable for at least 3 weeks after injection. Histological examination of muscles injected with beta-galactosidase-expressing C2C12 myoblasts demonstrated that many of the injected cells had fused to form multinucleated myotubes. Thus, genetically engineered myoblasts can be used for the stable delivery of recombinant proteins into the circulation.


Cross-Modal Correspondences Enhance Performance on a Colour-to-Sound Sensory Substitution Device.
Visual sensory substitution devices (SSDs) can represent visual characteristics through distinct patterns of sound, allowing a visually impaired user access to visual information. Previous SSDs have avoided colour and when they do encode colour, have assigned sounds to colour in a largely unprincipled way. This study introduces a new tablet-based SSD termed the 'Creole' (so called because it combines tactile scanning with image sonification) and a new algorithm for converting colour to sound that is based on established cross-modal correspondences (intuitive mappings between different sensory dimensions). To test the utility of correspondences, we examined the colour–sound associative memory and object recognition abilities of sighted users who had their device either coded in line with or opposite to sound–colour correspondences. Improved colour memory and reduced colour-errors were made by users who had the correspondence-based mappings. Interestingly, the colour–sound mappings that provided the highest improvements during the associative memory task also saw the greatest gains for recognising realistic objects that also featured these colours, indicating a transfer of abilities from memory to recognition. These users were also marginally better at matching sounds to images varying in luminance, even though luminance was coded identically across the different versions of the device. These findings are discussed with relevance for both colour and correspondences for sensory substitution use.


The prognostic value of pre-treatment thrombocytosis in two cohorts of patients with non-small cell lung cancer treated with curatively intended chemoradiotherapy.
Chemoradiotherapy is the standard of care for inoperable stage III non-small cell lung cancer (NSCLC). This treatment, however, offers only a small chance of cure and is associated with many side effects. Little research has been made concerning which patients benefit most/least from the treatment. The present study evaluates the prognostic value of anemia, leukocytosis and thrombocytosis at diagnosis in this treatment setting. In the present study, data were collected retrospectively for 222 patients from two different phase II studies conducted between 2002-2007 in Sweden with patients treated with chemoradiotherapy for stage IIIA-IIIB NSCLC. Clinical data and the serum values of hemoglobin (Hgb), White blood cells (WBC) and Platelets (Plt) at enrollment were collected for all patients and studied in relation to overall survival using Kaplan-Meier product-limit estimates and a multivariate Cox proportional hazards regression model. The results showed that patients with thrombocytosis (Plt > 350 x 109/L) had a shorter median overall survival (14.5 months) than patients with normal Plt at baseline (23.7 months). Patients with leukocytosis (WBC > 9 x 109/L) had a shorter median survival (14.9 months) than patients with a normal WBC at baseline (22.5 months). However, in a multivariate model including all lab parameters and clinical factors, only thrombocytosis and performance status displayed a prognostic significance. In Conclusion, thrombocytosis showed to be an independent prognostic marker associated with shorter overall survival in stage III NSCLC treated with curatively intended chemoradiotherapy. This knowledge can potentially be used together with established prognostic factors, such as performance status when choosing the optimal therapy for the individual patient in this clinical setting.


Improving BiVO4 photoanodes for solar water splitting through surface passivation.
BiVO4 has shown great potential as a semiconductor photoanode for solar water splitting. Significant improvements made during recent years allowed researchers to obtain a photocurrent density of up to 4.0 mA cm(-2) (AM1.5 sunlight illumination, 1.23 VRHE bias). For further improvements of the BiVO4 photoelectrodes, a deep understanding of the processes occurring at the BiVO4-H2O interface is crucial. Employing an electrochemical loading and removal process of NiOx, we show here that carrier recombination at this interface strongly affects the photocurrents. The removal of NiOx species by electrochemical treatment in a phosphate electrolyte leads to significantly increased photocurrents for BiVO4 photoelectrodes. At a bias of 1.23 VRHE, the Incident Photon-to-Current Efficiency (IPCE) at 450 nm reaches 43% for the passivated BiVO4 electrode under back side illumination. A model incorporating heterogeneity of NiOx centers on the BiVO4 surface (OER catalytic centers, recombination centers, and passivation centers) is proposed to explain this improved performance.


Radiographic and clinical outcomes following etanercept monotherapy in Japanese methotrexate-naïve patients with active rheumatoid arthritis.
Objectives: Compare outcomes with methotrexate (MTX) or etanercept (ETN) monotherapy in Japanese patients with active rheumatoid arthritis (RA) who were MTX-naïve or with intolerance or inadequate response to prior MTX (MTX-IR).Methods: Post hoc analysis of a phase 3 study comparing MTX, ETN 10 mg twice weekly, and ETN 25 mg twice weekly in Japanese patients with RA. Disease activity was evaluated using American College of Rheumatology (ACR) scores and 28-joint Disease Activity Score (DAS28), radiographic progression evaluated using van der Heijde's modified Total Sharp Score (mTSS), and functional status evaluated using Health Assessment Questionnaire Disability Index (HAQ-DI).Results: Among MTX-naïve and MTX-IR patients, greater proportions of those randomized to either ETN group achieved ACR20, ACR50, ACR70, DAS28 ≤3.2 or <2.6, clinically relevant inhibition of mTSS changes, and reductions in HAQ-DI compared with MTX at the majority of time points. There were very few clinically meaningful differences between ETN groups for any of the variables evaluated.Conclusion: ETN monotherapy was more effective than MTX in both MTX-naïve and MTX-IR patients, with very few clinically meaningful differences between ETN 10 mg and ETN 25 mg when given twice weekly. The relative benefits of ETN were greater in MTX-naïve patients than MTX-IR patients.ClinicalTrials.gov identifierNCT00445770.


Case Report: A Cluster of Plasmodium falciparum Malaria Cases among Thai Workers in Gembu, Nigeria.
Although falciparum malaria is an important risk among travelers to sub-Saharan Africa, many travelers remain unaware of this risk. In October 2015, we found a cluster of imported Plasmodium falciparum malaria cases among Thai gem mine workers in Nigeria; none had received malaria chemoprophylaxis or information regarding malaria risk. The index case developed fever and visited our hospital on arrival day in Thailand after his 3-week stay in Nigeria. Plasmodium falciparum was found in his blood. He recovered completely 3 days post-admission. After we requested he contact his colleagues in Nigeria regarding malaria risk, we found that three of his five colleagues currently had fever, were diagnosed with malaria, and were being treated in a local hospital. Two were successfully treated in Nigeria. Although their blood films were negative for malaria, we could confirm that they recently had malaria because the polymerase chain reaction (PCR) was still positive for P. falciparum. Tragically, the last febrile case died in Nigeria 6 days post-admission, after developing jaundice and alteration of consciousness. The two colleagues without fever symptoms were also tested by PCR, which was negative for malaria. In conclusion, we found that four of six workers had malaria in this cluster, which was equal to 66.7% attack rate. There is an urgent need to raise awareness of malaria among workers in highly endemic areas. Clinical practice for travelers who are ill on their return should not only focus on individual cases but also consider potential disease clusters.


1970s and 'Patient 0' HIV-1 genomes illuminate early HIV/AIDS history in North America.
The emergence of HIV-1 group M subtype B in North American men who have sex with men was a key turning point in the HIV/AIDS pandemic. Phylogenetic studies have suggested cryptic subtype B circulation in the United States (US) throughout the 1970s and an even older presence in the Caribbean. However, these temporal and geographical inferences, based upon partial HIV-1 genomes that postdate the recognition of AIDS in 1981, remain contentious and the earliest movements of the virus within the US are unknown. We serologically screened >2,000 1970s serum samples and developed a highly sensitive approach for recovering viral RNA from degraded archival samples. Here, we report eight coding-complete genomes from US serum samples from 1978-1979-eight of the nine oldest HIV-1 group M genomes to date. This early, full-genome 'snapshot' reveals that the US HIV-1 epidemic exhibited extensive genetic diversity in the 1970s but also provides strong evidence for its emergence from a pre-existing Caribbean epidemic. Bayesian phylogenetic analyses estimate the jump to the US at around 1970 and place the ancestral US virus in New York City with 0.99 posterior probability support, strongly suggesting this was the crucial hub of early US HIV/AIDS diversification. Logistic growth coalescent models reveal epidemic doubling times of 0.86 and 1.12 years for the US and Caribbean, respectively, suggesting rapid early expansion in each location. Comparisons with more recent data reveal many of these insights to be unattainable without archival, full-genome sequences. We also recovered the HIV-1 genome from the individual known as 'Patient 0' (ref. 5) and found neither biological nor historical evidence that he was the primary case in the US or for subtype B as a whole. We discuss the genesis and persistence of this belief in the light of these evolutionary insights.


A comparison of asthma deaths and near-fatal asthma attacks in South Australia.
Studies seeking to identify factors predictive of asthma mortality have relied on information obtained from relatives, other close acquaintances, and doctors who cared for the deceased. We wanted to determine whether asthmatics who have suffered a near-fatal asthma attack (NFA) are similar to asthmatics who have died of asthma with respect to important features, because studies of NFA asthmatics may provide a better insight into causes of asthma death. Such studies would avoid the difficulties associated with seeking information secondhand from proxy informants. Two groups were studied: asthmatics who had suffered a near-fatal asthma attack resulting in a visit to the accident and emergency departments of teaching hospitals (n = 154), and asthmatics certified as dying of asthma who, following panel review, were confirmed to have died from this disease (n = 80). For each case in the two groups, an interview questionnaire was administered to a close acquaintance (household or family member) and to the general practitioner. Both groups shared many important characteristics. Similarities related to: frequency of symptoms; frequency of hospital and intensive care unit admissions for asthma; use of asthma crisis plans; compliance with prescribed medications; quality of personal asthma management; and asthma severity. The two groups also showed similar psychiatric profiles, and similar use of asthma medications on a regular basis and with increased symptoms. However, NFA cases tended to be younger, were more likely to be male, and less likely to have concurrent medical conditions.(ABSTRACT TRUNCATED AT 250 WORDS)


Polycaprolactone- and polycaprolactone/ceramic-based 3D-bioplotted porous scaffolds for bone regeneration: A comparative study.
One of the critical challenges that scaffolding faces in the organ and tissue regeneration field lies in mimicking the structure, and the chemical and biological properties of natural tissue. A high-level control over the architecture, mechanical properties and composition of the materials in contact with cells is essential to overcome such challenge. Therefore, definition of the method, materials and parameters for the production of scaffolds during the fabrication stage is critical. With the recent emergence of rapid prototyping (RP), it is now possible to create three-dimensional (3D) scaffolds with the essential characteristics for the proliferation and regeneration of tissues, such as porosity, mechanical strength, pore size and pore interconnectivity, and biocompatibility. In this study, we employed 3D bioplotting, a RP technology, to fabricate scaffolds made from (i) pure polycaprolactone (PCL) and (ii) a composite based on PCL and ceramic micro-powder. The ceramics used for the composite were bovine bone filling Nukbone® (NKB), and hydroxyapatite (HA) with 5%, 10% or 20% wt. The scaffolds were fabricated in a cellular lattice structure (i.e. meshing mode) using a 0/90° lay down pattern with a continuous contour filament in order to achieve interconnected porous reticular structures. We varied the temperature, as well as injection speed and pressure during the bioplotting process to achieve scaffolds with pore size ranging between 200 and 400μm and adequate mechanical stability. The resulting scaffolds had an average pore size of 323μm and an average porosity of 32%. Characterization through ATR-FTIR revealed the presence of the characteristic bands of hydroxyapatite in the PCL matrix, and presented an increase of the intensity of the phosphate and carbonyl bands as the ceramic content increased. The bioplotted 3D scaffolds have a Young's modulus (E) in the range between 0.121 and 0.171GPa, which is compatible with the modulus of natural bone. PCL/NKB scaffolds, particularly 10NKBP (10% NKB wt.) exhibited the highest proliferation optical density, demonstrating an evident osteoconductive effect when cultured in Dulbecco's Modified Eagle Medium (DMEM). Scanning electron microscopy (SEM) confirmed osteoblast anchorage to all composite scaffolds, but a low adhesion to the all-PCL scaffold, as well as cell proliferation. The results from this study demonstrate the potential of PCL/NKB 3D bioplotted scaffolds as viable platforms to enable osseous tissue formation, which can be used in several tissue engineering applications, including improvement of bone tissue regeneration.


Imaging ellipsometry based method and algorithm for the analysis of fiber-fiber bonds in a paper network.
The measurement of the bonded area of pulp fibers has been an unsolved issue in paper science for more than 40 years. By the use of an established pulp fiber model, and a 4 × 4 transfer matrix formalism we simulated the optical behavior of pulp fibers in a modified imaging ellipsometer, and we demonstrate that there are rather strong symmetries in the ellipsometric angles Ψ and Δ when comparing single fibers, unbonded fiber crossings, and fiber-fiber bonds. Based on these symmetries we propose and test an algorithm that allows to distinguish the three cases (single fibers, unbonded fiber crossings, and fiber-fiber bonds) in the analysis of ellipsometric data.