First Report of Iris yellow spot virus Infecting Onion in Bosnia and Herzegovina.
In July 2012, a survey was conducted to determine the presence of tospoviruses in Bosnia and Herzegovina, symptoms resembling those caused by Iris yellow spot virus (IYSV; genus Tospovirus, family Bunyaviridae) were observed in an onion (Allium cepa) seed crop in the Gornji Karajzovci locality (Region of Banja Luka). Symptoms included chlorotic to necrotic, straw-colored, spindle- and diamond-shaped lesions, variable in size and randomly distributed on the leaves and particularly on the scapes. Later the lesions enlarged and coalesced, causing scape breakage. Affected plants occurred throughout the field and disease incidence was estimated at 20%. Symptomatic plants were sampled and assayed by double-antibody sandwich (DAS)-ELISA test using commercial polyclonal antisera (Bioreba AG, Reinach, Switzerland) against IYSV and two other tospoviruses, Tomato spotted wilt virus (TSWV) and Impatiens necrotic spot virus (INSV). Commercial positive and negative controls were included in each test. IYSV was detected serologically in 19 of 20 screened samples and none of the samples tested positive for TSWV or INSV. The virus was mechanically transmitted from an ELISA-positive sample (302-12) to five of each Petunia × hybrida and Nicotiana benthamiana using chilled 0.01 M phosphate buffer (pH 7) containing 0.1% sodium sulfite (1). All inoculated P. × hybrida showed local necrotic spots, while N. benthamiana developed mild mosaic 4 and 10 days post-inoculation, respectively. However, difficulties were encountered in reproducing the disease symptoms on mechanically inoculated onion plants corroborating a previous study (2). Serological findings were verified with reverse transcription (RT)-PCR. Total RNAs from all naturally infected onion plants as well as mechanically infected N. benthamiana plants were extracted with the RNease Plant Mini Kit (Qiagen, Hilden, Germany). RT-PCR was performed with One-Step RT-PCR Kit (Qiagen) using IYSV-specific primers IYSV56U/IYSV917L (3), designed to amplify an 896-bp fragment of the S RNA which includes whole nucleocapsid (N) gene. Total RNAs from Serbian IYSV isolate from onion (GenBank Accession No. EU586203) and from healthy onion plants were used as positive and negative controls, respectively. An amplicon of the expected size was obtained from each of the plants assayed as well as from positive control, but not from the negative control. The amplified products derived from onion isolate 302-12 was purified (QIAquick PCR Purification Kit, Qiagen), sequenced directly (JX861126), and compared with known IYSV isolates. Sequence analysis of the complete N gene, conducted with MEGA5 software (4), revealed the highest nucleotide identity of 99.5% (100% amino acid identity) with IYSV onion isolate (DQ658242) from Texas. To our knowledge, this is the first report of IYSV in Bosnia and Herzegovina. Onion is an important and traditionally grown vegetable crop in Bosnia and Herzegovina and the presence of IYSV could represent an important constraint to onion and other susceptible host production. The discovery of IYSV on onion should prompt more detailed surveys, thorough inspections and subsequent testing to establish the distribution and incidence of IYSV in Bosnia and Herzegovina. References: (1) A. Kritzman et al. Plant Dis. 85:838, 2001. (2) L. Pozzer et al. Plant Dis. 83:345, 1999. (3) I. Robène-Soustrade et al. Plant Pathol. 55:288, 2006. (4) K. Tamura et al. Mol. Biol. Evol. 28:2731, 2011.


Sinoatrial transmission and atrial invasion during normal rhythm in the rabbit heart.
Intracellular microelectrodes and small unipolar leads applied to endocardial surface of the right atrium in vitro were used to study the complex extracellular wave patterns recorded from the neighborhood of the cardiac pacemaker during spontaneous activity. Sinus activity propagated slowly toward the site of atrial invasion on the venous border of the crista terminalis. Atrial activation was marked by a primary negative wave that appeared 20-40 msec after pacemaker firing. Two sources of complex multiphasic waves were found. First, potentials from transitional sinus tissue propagated toward the atrium and caused low-voltage waves that preceded and slurred the onset of the atrial initial negativity. Second, bundles and layers of the crista terminalis muscle were excited asynchronously around the invasion region, as if cross-connections were infrequent. Waves originating from this source occurred after the firing of the invasion site. No extracellular wave could be associated with the firing of the true pacemaker cells. The sinoatrial ring bundle (SARB) yielded a discrete biphasic deflection along most of its way toward the coronary sinus. This potential appeared most frequently after that of the adjoining cristal muscle, raising questions about the functional role of the SARB as an internodal preferential pathway.


Analysis of the rat hypothalamus proteome by data-independent label-free LC-MS/MS.
Studies of neuronal, endocrine, and metabolic disorders would be facilitated by characterization of the hypothalamus proteome. Protein extracts prepared from 16 whole rat hypothalami were measured by data-independent label-free nano LC-MS/MS. Peptide features were detected, aligned, and searched against a rat Swiss-Prot database using ProteinLynx Global Server v.2.5. The final combined dataset comprised 21 455 peptides, corresponding to 622 unique proteins, each identified by a minimum of two distinct peptides. The majority of the proteins (69%) were cytosolic, and 16% were membrane proteins. Important proteins involved in neurological and synaptic function were identified including several members of the Ras-related protein family and proteins involved in glutamate biosynthesis.


Comparative evaluation of molecular diagnostic tests for Nucleospora salmonis and prevalence in migrating juvenile salmonids from the Snake River, USA.
Nucleospora salmonis is an intranuclear microsporidian that primarily infects lymphoblast cells and contributes to chronic lymphoblastosis and a leukemia-like condition in a range of salmonid species. The primary goal of this study was to evaluate the prevalence of N. salmonis in out-migrating juvenile hatchery and wild Chinook salmon Oncorhynchus tshawytscha and steelhead O. mykiss from the Snake River in the U.S. Pacific Northwest. To achieve this goal, we first addressed the following concerns about current molecular diagnostic tests for N. salmonis: (1) nonspecific amplification patterns by the published nested polymerase chain reaction (nPCR) test, (2) incomplete validation of the published quantitative PCR (qPCR) test, and (3) whether N. salmonis can be detected reliably from nonlethal samples. Here, we present an optimized nPCR protocol that eliminates nonspecific amplification. During validation of the published qPCR test, our laboratory developed a second qPCR test that targeted a different gene sequence and used different probe chemistry for comparison purposes. We simultaneously evaluated the two different qPCR tests for N. salmonis and foundthat both assays were highly specific, sensitive, and repeatable. The nPCR and qPCR tests had good overall concordance when DNA samples derived from both apparently healthy and clinically diseased hatchery rainbow trout were tested. Finally, we demonstrated that gill snips were a suitable tissue for nonlethal detection of N. salmonis DNA in juvenile salmonids. Monitoring of juvenile salmonid fish in the Snake River over a 3-year period revealed low prevalence of N. salmonis in hatchery and wild Chinook salmon and wild steelhead but significantly higher prevalence in hatchery-derived steelhead. Routine monitoring of N. salmonis is not performed for all hatchery steelhead populations. At present, the possible contribution of this pathogen to delayed mortality of steelhead has not been determined.


Effect of individual omega-3 fatty acids on the risk of prostate cancer: a systematic review and dose-response meta-analysis of prospective cohort studies.
Epidemiological studies have suggested inconsistent associations between omega-3 polyunsaturated fatty acids (n-3 PUFAs) and prostate cancer (PCa) risk. We performed a dose-response meta-analysis of prospective observational studies investigating both dietary intake and circulating n-3 PUFAs and PCa risk. PubMed and EMBASE prior to February 2014 were searched, and 16 publications were eligible. Blood concentration of docosahexaenoic acid, but not alpha-linolenic acid or eicosapentaenoic acid, showed marginal positive association with PCa risk (relative risk for 1% increase in blood docosahexaenoic acid concentration: 1.02; 95% confidence interval, 1.00-1.05; I(2) = 26%; P = 0.05 for linear trend), while dietary docosahexaenoic acid intake showed a non-linear positive association with PCa risk (P < 0.01). Dietary alpha-linolenic acid was inversely associated with PCa risk (relative risk for 0.5 g/day increase in alpha-linolenic acid intake: 0.99; 95% confidence interval, 0.98-1.00; I(2) = 0%; P = 0.04 for linear trend), which was dominated by a single study. Subgroup analyses indicated that blood eicosapentaenoic acid concentration and blood docosahexaenoic acid concentration were positively associated with aggressive PCa risk and nonaggressive PCa risk, respectively. Among studies with nested case-control study designs, a 0.2% increase in blood docosapentaenoic acid concentration was associated with a 3% reduced risk of PCa (relative risk 0.97; 95% confidence interval, 0.94-1.00; I(2) = 44%; P = 0.05 for linear trend). In conclusion, different individual n-3 PUFA exposures may exhibit different or even opposite associations with PCa risk, and more prospective studies, especially those examining dietary n-3 PUFAs and PCa risk stratified by severity of cancer, are needed to confirm the results.


Systems biology studies of adult paragonimus lung flukes facilitate the identification of immunodominant parasite antigens.
Paragonimiasis is a food-borne trematode infection acquired by eating raw or undercooked crustaceans. It is a major public health problem in the far East, but it also occurs in South Asia, Africa, and in the Americas. Paragonimus worms cause chronic lung disease with cough, fever and hemoptysis that can be confused with tuberculosis or other non-parasitic diseases. Treatment is straightforward, but diagnosis is often delayed due to a lack of reliable parasitological or serodiagnostic tests. Hence, the purpose of this study was to use a systems biology approach to identify key parasite proteins that may be useful for development of improved diagnostic tests. The transcriptome of adult Paragonimus kellicotti was sequenced with Illumina technology. Raw reads were pre-processed and assembled into 78,674 unique transcripts derived from 54,622 genetic loci, and 77,123 unique protein translations were predicted. A total of 2,555 predicted proteins (from 1,863 genetic loci) were verified by mass spectrometric analysis of total worm homogenate, including 63 proteins lacking homology to previously characterized sequences. Parasite proteins encoded by 321 transcripts (227 genetic loci) were reactive with antibodies from infected patients, as demonstrated by immunoaffinity purification and high-resolution liquid chromatography-mass spectrometry. Serodiagnostic candidates were prioritized based on several criteria, especially low conservation with proteins in other trematodes. Cysteine proteases, MFP6 proteins and myoglobins were abundant among the immunoreactive proteins, and these warrant further study as diagnostic candidates. The transcriptome, proteome and immunolome of adult P. kellicotti represent a major advance in the study of Paragonimus species. These data provide a powerful foundation for translational research to develop improved diagnostic tests. Similar integrated approaches may be useful for identifying novel targets for drugs and vaccines in the future.


Analysis of brain activation patterns using a 3-D scale-space primal sketch.
A fundamental problem in brain imaging concerns how to define functional areas consisting of neurons that are activated together as populations. We propose that this issue can be ideally addressed by a computer vision tool referred to as the scale-space primal sketch. This concept has the attractive properties that it allows for automatic and simultaneous extraction of the spatial extent and the significance of regions with locally high activity. In addition, a hierarchical nested tree structure of activated regions and subregions is obtained. The subject in this article is to show how the scale-space primal sketch can be used for automatic determination of the spatial extent and the significance of rCBF changes. Experiments show the result of applying this approach to functional PET data, including a preliminary comparison with two more traditional clustering techniques. Compared to previous approaches, the method overcomes the limitations of performing the analysis at a single scale or assuming specific models of the data.


Interactions between Clotrimazole and selected essential oils against Malassezia pachydermatis clinical isolates.
The aim of this study was to investigate interactions between conventional antifungal drug and essential oils against isolates of Malassezia pachydermatis. Antifungal activity of Cinnamomum cassia, Melaleuca alternifolia, Mentha piperita, Origanum vulgare and Syzygium aromaticum essential oils were tested against 19 strains of M. pachydermatis isolated from healthy dogs and reference strain M. pachydermatis CBS 1879. The checkerboard assay was used to search for in- teractions. Synergism was observed for the combination of clotrimazole with Melaleuca alternifolia essential oil, Mentha piperita and Origanum vulgare. The combinations of Cinnamomum cassia and Syzygium aromaticum essential oils with clotrimazole showed indifferent effect. Additive antimicrobial activity was observed for the combination of clotrimazole with Syzygium aromaticum and Melaleuca alternifolia essential oils against reference strain. The obtained results showed synergistic interactions between essential oils and clotrimazole which could improve effectiveness of this antifungal drug.


Pediatric end-stage renal disease: single center analysis.
Data describing end stage renal disease in Jordan is very limited, due to the absence of internal center as well as national registry systems. In this retrospective analysis, we define the etiology, prevalence, incidence as well as other demographic features of pediatric end stage renal disease and renal replacement therapy at King Hussein Medical Center. All children who entered the chronic dialysis program in our center from January 2001 to December 2005 were included in the study. Children who were transplanted pre-emptively were also included. A total number of 42 patients were included. The mean age at time of dialysis initiation was 11.10+/-2.25 years; 19 (45.2%) were males. The prevalence of ESRD in Jordan children was calculated to be 14.5 patients per million. Hemodialysis (HD) was the primary modality of therapy in 40 (95.2%) patients. The most common cause of ESRD in our children was acquired glomerulopathy in 13 (31%) patients, followed by oxalosis in seven (16.7%), and neurogenic bladder in six (14.3 %). Thirteen patients were transplanted; the one and three year graft survival rates were 87.5% and 72.5%, respectively. We conclude that there are some peculiar features for pediatric ESRD in Jordan including the high incidence of oxalosis and neurogenic bladder. A national registry for children with ESRD should be established as this may have serious implications on the choice of renal replacement therapy.


Monitoring Solar Radiation UV Exposure in the Comoros.
As part of the UV-Indien project, a station for measuring ultraviolet radiation and the cloud fraction was installed in December 2019 in Moroni, the capital of the Comoros, situated on the west coast of the island of Ngazidja. A ground measurement campaign was also carried out on 12 January 2020 during the ascent of Mount Karthala, located in the center of the island of Ngazidja. In addition, satellite estimates (Ozone Monitoring Instrument and TROPOspheric Monitoring Instrument) and model outputs (Copernicus Atmospheric Monitoring Service and Tropospheric Ultraviolet Model) were combined for this same region. On the one hand, these different measurements and estimates make it possible to quantify, evaluate, and monitor the health risk linked to exposure to ultraviolet radiation in this region, and, on the other, they help to understand how cloud cover influences the variability of UV-radiation on the ground. The measurements of the Ozone Monitoring Instrument onboard the EOS-AURA satellite, being the longest timeseries of ultraviolet measurements available in this region, make it possible to quantify the meteorological conditions in Moroni and to show that more than 80% of the ultraviolet indices are classified as high and that 60% of these are classified as extreme. The cloud cover measured in Moroni by an All Sky Camera was used to distinguish between the cases of UV index measurements taken under clear or cloudy sky conditions. The ground-based measurements thus made it possible to describe the variability of the diurnal cycle of the UV index and the influence of cloud cover on this parameter. They also permitted the satellite measurements and the results of the simulations to be validated. In clear sky conditions, a relative difference of between 6 and 11% was obtained between satellite or model estimates and ground measurements. The ultraviolet index measurement campaign on Mount Karthala showed maximum one-minute standard erythemal doses at 0.3 SED and very high daily cumulative erythemal doses at more than 80 SED. These very high levels are also observed throughout the year and all skin phototypes can exceed the daily erythemal dose threshold at more than 20 SED.


Flavonoids protect neurons from oxidized low-density-lipoprotein-induced apoptosis involving c-Jun N-terminal kinase (JNK), c-Jun and caspase-3.
Oxidative stress has been associated with neuronal loss in neurodegenerative diseases and during age-associated cognitive decline. Flavonoids have been proposed to play a useful role in protecting the central nervous system against oxidative and excitotoxic stress, although the mechanism of action is unknown. Using oxidized low-density lipoprotein (oxLDL) as the oxidative insult we investigated the mechanism of neurotoxicity and attempted to identify possible sites of action of two of the most potent protective flavonoids, epicatechin and kaempferol, in cultured primary neurons. Using cultured striatal neurons and selective phosphospecific antibodies we addressed the potential role of extracellular signal-regulated kinases 1/2 (ERK1/2) and c-Jun N-terminal kinase (JNK). OxLDL stimulated a Ca(2+)-dependent activation of both ERK1/2 and JNK that was strongly inhibited by pre-treatment with low micromolar concentrations of epicatechin. Neurotoxicity induced by oxLDL, however, was neither reduced nor enhanced by inhibiting ERK1/2 activation with mitogen-activated protein kinase kinase (MEK) inhibitors, suggesting that this cascade is unlikely to be involved in either oxLDL toxicity or the protective effects of flavonoids. oxLDL caused a sustained activation of JNK that resulted in the phosphorylation of the transcription factor c-Jun, which was abolished in neurons pre-treated with flavonoids. Furthermore, oxLDL induced the cleavage of procaspase-3 and increased caspase-3-like protease activity in neurons, an effect which was strongly inhibited by pre-exposure to either epicatechin or kaempferol. In addition, a caspase-3 inhibitor reduced oxLDL-induced neuronal death, implicating an apoptotic mechanism. A major in vivo metabolite of epicatechin, 3'-O-methyl-epicatechin was as effective as epicatechin in protecting neurons. Thus dietary flavonoids might have potential as protective agents against neuronal apoptosis through selective actions within stress-activated cellular responses, including protein kinase signalling cascades.


Expression of TGF-beta isoforms during first trimester human embryogenesis.
We have studied the expression of the genes encoding transforming growth factors (TGFs) beta 1, beta 2 and beta 3 in human embryos ranging from 32 to 57 days post-coitum, using in situ hybridization. The spatial and temporal pattern of expression of each gene is distinct, though each occasionally overlaps. TGF-beta 1 is expressed in haematopoietic, endothelial and osteogenic tissues. TGF-beta 2 and TGF-beta 3 are expressed in a wide variety of mesenchymal tissues including areas of chondrogenic activity. TGF-beta 2 is also found in several epithelial and in the ventral nervous system. The differential transcript distributions are broadly similar to those seen in mouse embryos suggesting that there is conservation of TGF-beta gene regulatory sequences and developmental function across this species boundary.


The hand of Homo naledi.
A nearly complete right hand of an adult hominin was recovered from the Rising Star cave system, South Africa. Based on associated hominin material, the bones of this hand are attributed to Homo naledi. This hand reveals a long, robust thumb and derived wrist morphology that is shared with Neandertals and modern humans, and considered adaptive for intensified manual manipulation. However, the finger bones are longer and more curved than in most australopiths, indicating frequent use of the hand during life for strong grasping during locomotor climbing and suspension. These markedly curved digits in combination with an otherwise human-like wrist and palm indicate a significant degree of climbing, despite the derived nature of many aspects of the hand and other regions of the postcranial skeleton in H. naledi.


Community-based learning enhances doctor retention.
Rural doctor shortage is a problem in many countries. Factors associated with doctor retention were reported such as colleagues, workload, accommodations, transportation, proximity of family and friends, incentives and career path. Rural background recruitment, increasing the quantity of doctor production to supply in rural and remote areas, and regulation are claimed to alleviate doctor scarcity in rural communities. Many programs have been developed, but an imbalance in physician distribution persists. Community-based learning (CBL) is recommended by the WHO to promote doctor retention. The longer contact time of CBL is practical, but it is uncertain that this results in greater retention. The objective of this study is to determine the association between contact time of CBL and retention of doctor with rural background recruitment. A cohort study was performed. The study population was 10,018 doctors graduated during 2001-2010 and followed up at least to 2014. Of the 10,018 physicians, 2098 doctors (21%) were recruited from rural backgrounds by the Collaborative Project to Increase Production of Rural Doctor (CPIRD). Contact time of CBL was calculated to the proportion of total curricular credit hours. The primary outcome was retention rate in government health-care system over 4 years. Statistical analysis was performed using multiple logistic regression. A total of 5774 doctors (57.6%) were retained in the government health-care system. Higher percentages of CPIRD doctors were retained than normal track (72.1% and 53.8%, P < 0.001), especially in rural hospitals (60.3% and 38.4%, P < 0.001). Based on univariate analysis, CBL was slightly higher in retention than resignation group with statistical significance (2.97% and 2.90%, P = 0.045). Multiple logistic regression results showed that CBL, graduate entry, and geographic location of workplace were significantly associated with retention. CBL can enhance doctor retention. It should incorporate meaningful experience such as rural exposure together with classroom teaching to focus concepts and integrating service to the community. Graduate entry and geographic location of workplace also have an impact on decision-making regarding retention.


The effect of breathing hyperoxic gas during simulated submarine escape on venous gas emboli and decompression illness.
Raised internal pressure in a distressed submarine rapidly increases the risk of decompression sickness (DCS) following submarine escape. The hypothesis that breathing a hyperoxic gas during escape may reduce the risk of DCS was tested using goats. Shallow air saturation and simulated submarine escape dives were carried out either singularly or in combination (saturation, escape, or saturation followed by escape) using air or 60% / 40% oxygen (O2) / nitrogen (N2) mixture as breathing gas during the escapes. Post-surfacing, animals were observed for signs of DCI and O2 toxicity. Precordial Doppler ultrasound was used to score venous gas emboli (VGE) using the Kisman Masurel (KM) scale. Following escape from 2.5 MPa, the rate at which VGE disappeared in the hyperoxic group (n = 8) was significantly faster(p < 0.05) than the air group (n = 7). One case of pulmonary barotrauma with arterial gas embolism occurred in the air group, but no cases of DCS were observed. After saturation at 0.18 MPa followed by escape from 2.5 MPa, DCS occurred in four of 15 animals in the air group and in two of 16 animals in the hyperoxic group. The rate of disappearance of VGE was significantly faster (p < 0.01) in the hyperoxic group. O2 toxicity was not discernible in any of the animals.


A new restricted access molecularly imprinted polymer capped with albumin for direct extraction of drugs from biological matrices: the case of chlorpromazine in human plasma.
A new restricted access molecularly imprinted polymer coated with bovine serum albumin (RAMIP-BSA) was developed, characterized, and used for direct analysis of chlorpromazine in human plasma samples. The RAMIP-BSA was synthesized using chlorpromazine, methacrylic acid, and ethylene glycol dimethacrylate as template, functional monomer, and cross-linker, respectively. Glycerol dimethacrylate and hydroxy methyl methacrylate were used to promote a hydrophilic surface (high density of hydroxyl groups). Afterward, the polymer was coated with BSA using glutaraldehyde as cross-linker, resulting in a protein chemical shield around it. The material was able to eliminate ca. 99% of protein when a 44-mg mL(-1) BSA aqueous solution was passed through it. The RAMIP-BSA was packed in a column and used for direct analysis of chlorpromazine in human plasma samples in an online column switching high-performance liquid chromatography system. The analytical calibration curve was prepared in a pool of human plasma samples with chlorpromazine concentrations ranging from 30 to 350 μg L(-1). The correlation coefficient obtained was 0.995 and the limit of quantification was 30 μg L(-1). Intra-day and inter-day precision and accuracy presented variation coefficients and relative errors lower than 15% and within -15 and 15%, respectively. The sample throughput was 3 h(-1) (sample preparation and chromatographic analysis steps) and the same RAMIP-BSA column was efficiently used for about 90 cycles.


Mechanism of activation of human peripheral blood NK cells at the single cell level by Echinacea water soluble extracts: recruitment of lymphocyte-target conjugates and killer cells and activation of programming for lysis.
Echinacea purpurea, a plant originally used by native Americans to treat respiratory infections, has also been shown to exert immunomodulatory activities both in vivo and in vitro. However, the mechanism underlying Echinacea-induced immunomodulation remains largely unknown. This study examined in vitro the effects of soluble extracts of E. purpurea on natural killer (NK) cells present in human peripheral blood mononuclear cells (PBMC). Flow cytometric methods were used to examine activation, cytotoxicity, NK-target binding, and killer cell frequency. Treatment of PBMC with Echinacea overnight resulted in the activation of CD69 expression and increase in mean fluorescence intensity in both the CD16+ and CD16+CD56+ NK subsets. However, the frequency of CD16+ cells was decreased as well as the mean fluorescence intensity was down-regulated. NK cytotoxicity was augmented 100% at the concentration of 0.1 microg/ml of Echinacea in a short time (4-h) assay. Examination at the single cell level revealed augmentation of the frequency of CD56+ NK-target conjugates and a plateau was reached after 30-60 min of incubation. Likewise, the frequency of CD56+ killer cells in the conjugates was also significantly increased by Echinacea. There was recruitment of non-conjugated CD56+ cells into CD16+ NK-target conjugates and activation of the NK-target non-killer conjugates into killer cells. These findings demonstrate that Echinacea extracts are potent activators of NK cytotoxicity. Echinacea augments the frequency of NK target conjugates and activates the programming for lysis of NK cells.


A family study of alcohol dependence: coaggregation of multiple disorders in relatives of alcohol-dependent probands.
Alcohol dependence tends to aggregate within families. We analyzed data from the family collection of the Collaborative Study on the Genetics of Alcoholism to quantify familial aggregation using several different criterion sets. We also assessed the aggregation of other psychiatric disorders in the same sample to identify areas of possible shared genetic vulnerability. Age-corrected lifetime morbid risk was estimated in adult first-degree relatives of affected probands and control subjects for selected disorders. Diagnostic data were gathered by semistructured interview (the Semi-Structured Assessment for the Genetics of Alcoholism), family history, and medical records. Rates of illness were corrected by validating interview and family history reports against senior clinicians' all sources best estimate diagnoses. Sex, ethnicity, comorbidity, cohort effects, and site of ascertainment were also taken into account. Including data from 8296 relatives of alcoholic probands and 1654 controls, we report lifetime risk rates of 28.8% and 14.4% for DSM-IV alcohol dependence in relatives of probands and controls, respectively; respective rates were 37.0% and 20.5% for the less stringent DSM-III-R alcohol dependence, 20.9% and 9.7% for any DSM-III-R diagnosis of nonalcohol nonnicotine substance dependence, and 8.1% and 5.2% for antisocial personality disorder. Rates of specific substance dependence were markedly increased in relatives of alcohol-dependent probands for cocaine, marijuana, opiates, sedatives, stimulants, and tobacco. Aggregation was also seen for panic disorder, obsessive-compulsive disorder, posttraumatic stress disorder, and major depression. The risk of alcohol dependence in relatives of probands compared with controls is increased about 2-fold. The aggregation of antisocial personality disorder, drug dependence, anxiety disorders, and mood disorders suggests common mechanisms for these disorders and alcohol dependence within some families. These data suggest new phenotypes for molecular genetic studies and alternative strategies for studying the heterogeneity of alcohol dependence.


Some behavioral effects of histamine H1 antagonists in squirrel monkeys.
Graded dose of the histamine H1 antagonists tripelennamine (0.1-3.0 mg/kg), promethazine (0.1-3.0 mg/kg), diphenhydramine (0.3-10.0 mg/kg) and chlorpheniramine (0.3-1.7 mg/kg) increased rates of nonsuppressed responding under a second-order schedule of food presentation to a maximum beyond which responding was increased less or decreased. In contrast, the H2 antagonist, cimetidine, had no effect or decreased responding at the highest doses studied (56.0-100.0 mg/kg). Intermediate doses of tripelennamine, diphenhydramine and promethazine also increased rates of food-maintained responding that were suppressed by electric shock. Maximal increases in rates of suppressed responding were comparable to those produced by effective doses of chlordiazepoxide (3.0-10.0 mg/kg). Under identical conditions, clozapine (0.1-1.0 mg/kg) increased responding to a lesser extent, and d-amphetamine (0.01-0.3 mg/kg) and cimetidine (3.0-100.0 mg/kg) either did not increase or, at the highest doses, only decreased rates of suppressed responding. Doses of tripelennamine and diphenhydramine that increased rates of nonsuppressed and suppressed responding also maintained self-administration in cocaine-trained squirrel monkeys under a second-order schedule of i.v. injection. Rates and patterns of responding maintained by tripelennamine and diphenhydramine were comparable to those maintained by cocaine and d-amphetamine under identical conditions. The results show that histamine H1 antagonists can have pronounced rate-increasing effects on nonsuppressed and suppressed behavior, and that they can serve as reinforcers in monkeys. These effects occur at doses that probably are greater than those required to saturate H1 sites of action in central nervous system and may not be mediated solely through histaminic mechanisms.


Assay and properties of rab6 interaction with dynein-dynactin complexes.
RAB GTPases help to maintain the fidelity of membrane trafficking events by recruiting cytosolic tethering and motility factors to vesicle and organelle membranes. In the case of Rab6, it recruits the dynein-dynaction complex to Golgi-associated vesicles via an adaptor protein of the Bicaudal-D family. Here we describe methods for the identification of Rab6-binding partners in cell extracts. We then focus on the biochemical analysis of interactions with the dynein-dynactin complex and the adaptor proteins Bicaudal-D1 and -D2. Standard protocols for yeast two-hybrid analysis, and biochemical assays for the analysis of the interactions between Rab6, Bicaudal-D, and the subunits of the dynein-dynactin complex are outlined.


Carnitine levels in valproic acid-treated psychiatric patients: a cross-sectional study.
Carnitine facilitates the transport of long-chain fatty acids across the mitochondria for beta oxidation, and the removal of potentially toxic acylcoenzyme-A metabolites from the inner aspect of mitochondrion as acylcarnitines. Previous studies suggest a significant decrease in carnitine concentrations and changes in the ratio of acylcarnitine to free carnitine in seizure-disoriented patients treated with valproic acid (VPA), which may lead to clinical manifestations of carnitine deficiency. This study sought to explore whether the same decrease in plasma free carnitine and increase in acylcarnitines are seen when VPA is used in the treatment of patients with psychiatric disease. Thirty psychiatric patients treated with VPA for at least six months were selected for the study and granted informed consent for participation. Exclusion criteria included liver disorder or pancreatitis, metabolic defects known to affect plasma carnitine levels, or noncompliance with VPA regimen. Plasma free carnitine, total carnitine, VPA, and amylase levels were determined, and liver function tests (LFTs) were performed. Pearson correlations were conducted between VPA levels, levels and ratios of carnitines, as well as LFTs and amylase levels. Plasma free and total carnitine levels were lower than the reported normal range for the laboratory performing the assay, and the ratio of acylcarnitine to free carnitine was increased. There was a significant positive correlation of VPA levels and acylcarnitine-free carnitine ratio, a trend toward significance between VPA levels and acylcarnitine levels, and a marginal negative correlation between VPA levels and free carnitine levels. VPA levels correlated also with several LFTs and acylcarnitine levels. Octanoyl carnitine and acylcarnitine levels, as well as acylcarnitine-free carnitine and octanoyl-free carnitine ratios, correlated significantly with amylase levels. Although the study was limited by a cross-sectional design without direct control comparison, the findings suggest that patients with various psychiatric conditions treated with polypharmacy that includes VPA may have lower plasma carnitine levels than would be expected in healthy controls.


Increasing recruitment contacts between generalist residents at the Medical College of Georgia and rural and underserved communities.
To increase the number of residents choosing to practice in rural and underserved areas, the Medical College of Georgia in 1994 created the Office of Recruitment and Retention (ORR) for generalist physicians as part of its participation in The Robert Wood Johnson Foundation's Generalist Physician Initiative. Its major purpose is to increase the contact between generalist residents and practice opportunities, especially those in rural and underserved areas. The office has helped residents better understand the resources available in rural settings and has helped these communities better understand the needs of graduates in generalist residency programs. It became a point of contact for residents and communities. It also organized on-campus "practice opportunity fairs" that permitted community representatives to meet formally with generalist residents to provide information on practice opportunities and community resources. The office organized practice management sessions and provided legal consultation to residents desiring to enter private practice in rural settings. This program has already been instrumental in furthering contact between generalist residents and the rural communities they selected as clinical practice sites during training. Although it is too early to know the impact of these activities, communities increasingly use the office to recruit generalist physicians, and residents use it to identify appropriate practice sites.


Murray Valley encephalitis in an adult traveller complicated by long-term flaccid paralysis: case report and review of the literature.
Murray Valley encephalitis (MVE) virus, a mosquito-borne flavivirus, is the most common cause of viral encephalitis in the tropical 'Top End' of northern Australia. Clinical encephalitis due to MVE virus has a mortality rate of approximately 30%, with a similar proportion of patients being left with significant neurological deficits. We report the case of a 25-year-old man from the UK who acquired MVE while travelling through northern Australia. He required prolonged admission to the Intensive Care Unit and several years later remains partly ventilator-dependent, with flaccid quadriparesis. To our knowledge, this is the first reported case of MVE virus-induced flaccid paralysis in an adult in northern Australia, although it is well described in children. Paralysis was thought to be due to anterior horn cell involvement in the spinal cord and extensive bilateral thalamic destruction, both of which are well recognised complications of infection with MVE virus. Cases of flaccid paralysis with similar pathology have been described following infection with the related flavivirus Japanese encephalitis virus as well as more recently with West Nile virus. Our case highlights the potential severity of flavivirus-induced encephalitis and the importance of avoiding mosquito bites while travelling through endemic areas.


Baroreceptor reflex in heart failure.
Congestive heart failure is a syndrome that is usually initiated by a reduction in pump function of the heart, i.e. a decrease in cardiac output. Initially, a reduction in cardiac output leads to unloading of baroreceptor reflex that, in turn, increases heart rate through vago-sympathetic mechanisms and total peripheral resistance via an increase in sympathetic outflow to vascular beds. In this review we are thinking on how baroreceptor reflex plays a role in the abnormal control of the circulation in heart failure. This review and our recent studies suggest that: (1) baroreceptor reflex is blunted in heart failure; (2) central angiotensin II and reactive oxygen species play an important role in blunted baroreceptor reflex; (3) cardiac sympathetic afferent stimulation and chemoreceptor reflex inhibit baroreceptor reflex; and (4) exercise training normalizes abnormal reflexes in the heart failure state.


Computer generated hologram with geometric occlusion using GPU-accelerated depth buffer rasterization for three-dimensional display.
We present a method of rapidly producing computer-generated holograms that exhibit geometric occlusion in the reconstructed image. Conceptually, a bundle of rays is shot from every hologram sample into the object volume. We use z buffering to find the nearest intersecting object point for every ray and add its complex field contribution to the corresponding hologram sample. Each hologram sample belongs to an independent operation, allowing us to exploit the parallel computing capability of modern programmable graphics processing units (GPUs). Unlike algorithms that use points or planar segments as the basis for constructing the hologram, our algorithm's complexity is dependent on fixed system parameters, such as the number of ray-casting operations, and can therefore handle complicated models more efficiently. The finite number of hologram pixels is, in effect, a windowing function, and from analyzing the Wigner distribution function of windowed free-space transfer function we find an upper limit on the cone angle of the ray bundle. Experimentally, we found that an angular sampling distance of 0.01 degrees for a 2.66 degrees cone angle produces acceptable reconstruction quality.


SARS-CoV-2 ORF3b Is a Potent Interferon Antagonist Whose Activity Is Increased by a Naturally Occurring Elongation Variant.
One of the features distinguishing SARS-CoV-2 from its more pathogenic counterpart SARS-CoV is the presence of premature stop codons in its ORF3b gene. Here, we show that SARS-CoV-2 ORF3b is a potent interferon antagonist, suppressing the induction of type I interferon more efficiently than its SARS-CoV ortholog. Phylogenetic analyses and functional assays reveal that SARS-CoV-2-related viruses from bats and pangolins also encode truncated ORF3b gene products with strong anti-interferon activity. Furthermore, analyses of approximately 17,000 SARS-CoV-2 sequences identify a natural variant in which a longer ORF3b reading frame was reconstituted. This variant was isolated from two patients with severe disease and further increased the ability of ORF3b to suppress interferon induction. Thus, our findings not only help to explain the poor interferon response in COVID-19 patients but also describe the emergence of natural SARS-CoV-2 quasispecies with an extended ORF3b gene that may potentially affect COVID-19 pathogenesis.


Epigenetic analysis of sporadic and Lynch-associated ovarian cancers reveals histology-specific patterns of DNA methylation.
Diagnosis and treatment of epithelial ovarian cancer is challenging due to the poor understanding of the pathogenesis of the disease. Our aim was to investigate epigenetic mechanisms in ovarian tumorigenesis and, especially, whether tumors with different histological subtypes or hereditary background (Lynch syndrome) exhibit differential susceptibility to epigenetic inactivation of growth regulatory genes. Gene candidates for epigenetic regulation were identified from the literature and by expression profiling of ovarian and endometrial cancer cell lines treated with demethylating agents. Thirteen genes were chosen for methylation-specific multiplex ligation-dependent probe amplification assays on 104 (85 sporadic and 19 Lynch syndrome-associated) ovarian carcinomas. Increased methylation (i.e., hypermethylation) of variable degree was characteristic of ovarian carcinomas relative to the corresponding normal tissues, and hypermethylation was consistently more prominent in non-serous than serous tumors for individual genes and gene sets investigated. Lynch syndrome-associated clear cell carcinomas showed the highest frequencies of hypermethylation. Among endometrioid ovarian carcinomas, lower levels of promoter methylation of RSK4, SPARC, and HOXA9 were significantly associated with higher tumor grade; thus, the methylation patterns showed a shift to the direction of high-grade serous tumors. In conclusion, we provide evidence of a frequent epigenetic inactivation of RSK4, SPARC, PROM1, HOXA10, HOXA9, WT1-AS, SFRP2, SFRP5, OPCML, and MIR34B in the development of non-serous ovarian carcinomas of Lynch and sporadic origin, as compared to serous tumors. Our findings shed light on the role of epigenetic mechanisms in ovarian tumorigenesis and identify potential targets for translational applications.


Could wastewater analysis be a useful tool for China? - A review.
Analysing wastewater samples is an innovative approach that overcomes many limitations of traditional surveys to identify and measure a range of chemicals that were consumed by or exposed to people living in a sewer catchment area. First conceptualised in 2001, much progress has been made to make wastewater analysis (WWA) a reliable and robust tool for measuring chemical consumption and/or exposure. At the moment, the most popular application of WWA, sometimes referred as sewage epidemiology, is to monitor the consumption of illicit drugs in communities around the globe, including China. The approach has been largely adopted by law enforcement agencies as a device to monitor the temporal and geographical patterns of drug consumption. In the future, the methodology can be extended to other chemicals including biomarkers of population health (e.g. environmental or oxidative stress biomarkers, lifestyle indicators or medications that are taken by different demographic groups) and pollutants that people are exposed to (e.g. polycyclic aromatic hydrocarbons, perfluorinated chemicals, and toxic pesticides). The extension of WWA to a huge range of chemicals may give rise to a field called sewage chemical-information mining (SCIM) with unexplored potentials. China has many densely populated cities with thousands of sewage treatment plants which are favourable for applying WWA/SCIM in order to help relevant authorities gather information about illicit drug consumption and population health status. However, there are some prerequisites and uncertainties of the methodology that should be addressed for SCIM to reach its full potential in China.


Aerosol-fluorescence spectrum analyzer: real-time measurement of emission spectra of airborne biological particles.
We have assembled an aerosol-fluorescence spectrum analyzer (AFS), which can measure the fluorescence spectra and elastic scattering of airborne particles as they flow through a laser beam. The aerosols traverse a scattering cell where they are illuminated with intense (50 kW/cm(2)) light inside the cavity of an argon-ion laser operating at 488 nm. This AFS can obtain fluorescence spectra of individual dye-doped polystyrene microspheres as small as 0.5 µm in diameter. The spectra obtained from microspheres doped with pink and green-yellow dyes are clearly different. We have also detected the fluorescence spectra of airborne particles (although not single particles) made from various biological materials, e.g., Bacillus subtilis spores, B. anthrasis spores, riboflavin, and tree leaves. The AFS may be useful in detecting and characterizing airborne bacteria and other airborne particles of biological origin.


Generation Scotland: the Scottish Family Health Study; a new resource for researching genes and heritability.
Generation Scotland: the Scottish Family Health Study aims to identify genetic variants accounting for variation in levels of quantitative traits underlying the major common complex diseases (such as cardiovascular disease, cognitive decline, mental illness) in Scotland. Generation Scotland will recruit a family-based cohort of up to 50,000 individuals (comprising siblings and parent-offspring groups) across Scotland. It will be a six-year programme, beginning in Glasgow and Tayside in the first two years (Phase 1) before extending to other parts of Scotland in the remaining four years (Phase 2). In Phase 1, individuals aged between 35 and 55 years, living in the East and West of Scotland will be invited to participate, along with at least one (and preferably more) siblings and any other first degree relatives aged 18 or over. The total initial sample size will be 15,000 and it is planned that this will increase to 50,000 in Phase 2. All participants will be asked to contribute blood samples from which DNA will be extracted and stored for future investigation. The information from the DNA, along with answers to a life-style and medical history questionnaire, clinical and biochemical measurements taken at the time of donation, and subsequent health developments over the life course (traced through electronic health records) will be stored and used for research purposes. In addition, a detailed public consultation process will begin that will allow respondents' views to shape and develop the study. This is an important aspect to the research, and forms the continuation of a long-term parallel engagement process. As well as gene identification, the family-based study design will allow measurement of the heritability and familial aggregation of relevant quantitative traits, and the study of how genetic effects may vary by parent-of-origin. Long-term potential outcomes of this research include the targeting of disease prevention and treatment, and the development of screening tools based on the new genetic information. This study approach is complementary to other population-based genetic epidemiology studies, such as UK Biobank, which are established primarily to characterise genes and genetic risk in the population.


The hormonal effects of Tribulus terrestris and its role in the management of male erectile dysfunction--an evaluation using primates, rabbit and rat.
Hormonal effects of Tribulus terrestris (TT) were evaluated in primates, rabbit and rat to identify its usefulness in the management of erectile dysfunction (ED). TT extract was administered intravenously, as a bolus dose of 7.5, 15 and 30 mg/kg, in primates for acute study. Rabbits and normal rats were treated with 2.5, 5 and 10mg/kg of TT extract orally for 8 weeks, for chronic study. In addition, castrated rats were treated either with testosterone cypionate (10mg/kg, subcutaneously; biweekly for 8 weeks) or TT orally (5mg/kg daily for 8 weeks). Blood samples were analyzed for testosterone (T), dihydrotestosterone (DHT) and dehydroepiandrosterone sulphate (DHEAS) levels using radioimmunoassay. In primates, the increases in T (52%), DHT (31%) and DHEAS (29%) at 7.5mg/kg were statistically significant. In rabbits, both T and DHT were increased compared to control, however, only the increases in DHT (by 30% and 32% at 5 and 10mg/kg) were statistically significant. In castrated rats, increases in T levels by 51% and 25% were observed with T and TT extract respectively that were statistically significant. TT increases some of the sex hormones, possibly due to the presence of protodioscin in the extract. TT may be useful in mild to moderate cases of ED.


Systems analysis at the molecular scale.
Problems involving physiochemical phenomena on both the microscopic and macroscopic scales often raise similar sets of generic issues and questions. The complexity of these problems is beginning to make inoperative the traditional intuition-based approaches to their analysis and solution. The common characteristics of large, multivariable, complex molecular systems call for a new, more systematic approach to guide theoretical and experimental efforts. With mathematical modeling becoming an essential ingredient in the studies, it is argued that molecular systems analysis and especially the systematic tools of sensitivity analyis can play an increasingly important role in understanding and finding solutions to complex, chemically based problems.


Ligament reconstruction with or without tendon interposition to treat primary thumb carpometacarpal osteoarthritis. Surgical technique.
Trapezial excision with ligament reconstruction and trapezial excision with ligament reconstruction combined with tendon interposition have proven to be highly effective techniques for treating primary osteoarthritis of the thumb carpometacarpal joint. To determine whether tendon interposition and proximal migration of the thumb metacarpal affect the objective and subjective outcomes, we compared the long-term outcomes of these two procedures performed in similar patient groups. Forty-three patients (fifty-two thumbs) were randomized to undergo either trapezial excision with ligament reconstruction or the same procedure combined with tendon interposition. Fifteen patients treated with ligament reconstruction (group I) and sixteen patients treated with the same procedure with concomitant tendon interposition (group II) were evaluated after a mean follow-up period of 48.2 months. The outcomes were assessed with the Buck-Gramcko score, with the total score calculated on the basis of the objective and subjective results. The ability to perform activities requiring use of the thumb and to return to work was analyzed as well. Radiographs were evaluated to determine the amount of proximal metacarpal migration at rest and under stress. Postoperatively, the mean total Buck-Gramcko score was rated as excellent in group I and as good in group II (p = 0.036). Group I had significantly better mean scores for palmar and radial abduction, cosmetic appearance, and willingness to undergo the surgery again under similar circumstances (p < 0.05). The mean scores for tip-pinch strength and the mean subjective scores for pain, strength, daily function, dexterity, and overall satisfaction did not differ significantly between the groups. Both groups had satisfactory results with regard to their performance of activities of daily living and their ability to return to work. With the numbers available, the amount of proximal metacarpal migration, at rest and under stress, did not differ significantly between the groups. Tendon interposition does not affect the outcome after the ligament reconstruction for the treatment of osteoarthritis of the thumb carpometacarpal joint. Furthermore, proximal migration of the thumb metacarpal does not appear to influence the functional outcome.


Performance Evaluation of a Generative Adversarial Network for Deblurring Mobile-phone Cervical Images.
Visual examination forms an integral part of cervical cancer screening. With the recent rise in smartphone-based health technologies, capturing cervical images using a smartphone camera for telemedicine and automated screening is gaining popularity. However, such images are highly prone to image corruption, typically out-of-focus target or camera shake blur. In this paper, we applied a generative adversarial network (GAN) to deblur mobile-phone cervical (MC) images, and we evaluate the deblur quality using various measures. Our evaluation process is three-fold: first, we calculate the peak signal to noise ratio (PSNR) and the structural similarity (SSIM) of a test dataset with ground truth availability. Next, we calculate the perception based image quality evaluator (PIQE) score of a test dataset without ground truth availability. Finally, we classify a dataset of blurred and the corresponding deblurred images into normal/abnormal MC images. The resulting change in classification accuracy was our final assessment. Our evaluation experiments show that deblurring of MC images can potentially improve the accuracy of both manual and automated cancerous lesion screening.


Comorbidity between major depression and alcohol use disorder from adolescence to adulthood.
Limited information exists regarding the long-term development of comorbidity between Major Depressive Disorder (MDD) and Alcohol Use Disorder (AUD; abuse/dependence). Using a representative prospective study, we examine multiple aspects pertaining to MDD+AUD comorbidity, with a focus on the relation between disorders across periods (adolescence, early adulthood, adulthood) and cumulative impairments by age 30. 816 participants were diagnostically interviewed at ages 16, 17, 24, and 30. Rates of comorbid MDD+AUD were low in adolescence (2%), but increased in early adulthood (11%) and adulthood (7%). Rates of cumulative comorbidity were elevated (21%). Most individuals with a history of MDD or AUD had the other disorder, except for women with MDD. Prospectively, adolescent AUD predicted early adult MDD, while early adult MDD predicted adult AUD. Compared to pure disorders, MDD+AUD was associated with higher risk of alcohol dependence, suicide attempt, lower global functioning, and life dissatisfaction. Lifetime rates of comorbid MDD+AUD were considerably higher than in cross-sectional studies. Comorbidity was partly explained by bidirectional and developmentally-specific associations and predicted selected rather than generalized impairments. Clinically, our findings emphasize the need to always carefully assess comorbidity in patients with MDD or AUD, taking into account concurrency and developmental timing.


Preconcentration of indapamide from human urine using molecularly imprinted solid-phase extraction.
A simple, sensitive, and selective molecularly imprinted solid-phase extraction and spectrophotometric method has been developed for the clean-up and preconcentration of indapamide from human urine. Molecularly imprinted polymers were prepared by a non-covalent imprinting approach using indapamide as a template molecule, 2-(trifluoromethyl) acrylic acid as a functional monomer, ethylene glycol dimethacrylate as a crosslinker, N,N-azobisisobutyronitrile as a thermal initiator and acetonitrile as a porogenic solvent. A non-imprinted polymer was also prepared in the same way, but in the absence of template. Molecularly imprinted polymer and non-imprinted polymer sorbents were dry-packed into solid-phase extraction cartridges. Eluates from cartridges were analyzed using a spectrophotometer for the determination of indapamide by referring to the calibration curve in the range 0.14-1.50 μg/mL. Preconcentration factor, limit of detection, and limit of quantification were 16.30, 0.025 μg/mL, and 0.075 μg/mL, respectively. A relatively high imprinting factor (9.3) was also achieved and recovery values for the indapamide spiked into human urine were in the range of 80.1-81.2%. In addition, relatively low within-day (0.17-0.42%) and between-day (1.1-1.4%) precision values were obtained as well. The proposed molecularly imprinted solid-phase extraction and spectrophotometric method was successfully applied to selective extraction, preconcentration, and determination of indapamide from human urine samples.


Knee effusion and reflex inhibition of the quadriceps. A bar to effective retraining.
In order to investigate the difficulty of quadriceps training in the presence of an effusion into the knee we examined 13 patients with chronic effusions by recording isometric muscle strength. Maximal strength was markedly lower in the presence of an effusion, and aspiration of the effusion produced a 13.6% increase in strength (p less than 0.01). A further, small increase of 8% was recorded after intra-articular lignocaine injection. Isometric strength and surface integrated EMG correlated well in six patients. Two reflex mechanisms seem to inhibit quadriceps innervation in the presence of a persistent knee effusion, one mediated by pressure sensitive receptors, the other still unknown. Joint aspiration and systemic or intra-articular anti-inflammatory drug treatments are advised before any programme of quadriceps training to allow maximum effects to be achieved.


Visual hallucinations in neurological and ophthalmological disease: pathophysiology and management.
Visual hallucinations are common in older people and are especially associated with ophthalmological and neurological disorders, including dementia and Parkinson's disease. Uncertainties remain whether there is a single underlying mechanism for visual hallucinations or they have different disease-dependent causes. However, irrespective of mechanism, visual hallucinations are difficult to treat. The National Institute for Health Research (NIHR) funded a research programme to investigate visual hallucinations in the key and high burden areas of eye disease, dementia and Parkinson's disease, culminating in a workshop to develop a unified framework for their clinical management. Here we summarise the evidence base, current practice and consensus guidelines that emerged from the workshop.Irrespective of clinical condition, case ascertainment strategies are required to overcome reporting stigma. Once hallucinations are identified, physical, cognitive and ophthalmological health should be reviewed, with education and self-help techniques provided. Not all hallucinations require intervention but for those that are clinically significant, current evidence supports pharmacological modification of cholinergic, GABAergic, serotonergic or dopaminergic systems, or reduction of cortical excitability. A broad treatment perspective is needed, including carer support. Despite their frequency and clinical significance, there is a paucity of randomised, placebo-controlled clinical trial evidence where the primary outcome is an improvement in visual hallucinations. Key areas for future research include the development of valid and reliable assessment tools for use in mechanistic studies and clinical trials, transdiagnostic studies of shared and distinct mechanisms and when and how to treat visual hallucinations.


Variables associated with general practitioners taking on serious mental disorder patients.
As part of community-based initiatives to strengthen integrated care and promote patient recovery, GPs are asked to play a greater part in treating serious mental disorder (SMD) patients. All current healthcare reforms favour the reinforcement of primary care. More information on enhancing the role of GPs in mental health would benefit policymakers, especially as regards SMD patients, where little research has been published as yet. This article assesses variables associated with GPs taking on SMD patients. The study, encompassing multiple sites, is based on a sample of 398 GPs, representative of the GP population in the Canadian province of Quebec. GPs were asked to answer a 143-item questionnaire on their socio-demographic and clinical practice profiles, patient characteristics, perceived inter-professional relationships and quality of care. Descriptive, bivariate and multivariate analyses were performed. Our data highlighted that GPs currently followed up only a minority of SMD patients on a continuous basis and far fewer for both physical and mental health problems. A linear regression model that accounts for 43% of the variance was generated. The best variables associated positively with GPs taking on SMD patients were: frequency of referrals for joint follow-up with other resources, and involvement in post-hospitalization follow-up. Conversely, lack of expertise in mental health (related in our model to frequency of mental disorder patient transfer due to insufficient mental health training) is associated with a lower incidence of GPs taking on patients. As advocated in current healthcare reforms, our study confirms the need to promote greater GP involvement in integrated care models and enhance their training in mental health--thereby helping to reverse the trend among GPs of transferring SMD patients to specialized care. Patients with stable SMDs ought to have the same care access as the general population.


Randomized Interventional Study on Prediction of Preeclampsia/Eclampsia in Women With Suspected Preeclampsia: INSPIRE.
The ratio of maternal serum sFlt-1 (soluble fms-like tyrosine kinase 1) to PlGF (placental growth factor) has been used retrospectively to rule out the occurrence of preeclampsia, a pregnancy hypertensive disorder, within 7 days in women presenting with clinical suspicion of preeclampsia. A prospective, interventional, parallel-group, randomized clinical trial evaluated the use of sFlt-1/PlGF ratio in women presenting with suspected preeclampsia. Women were assigned to reveal (sFlt-1/PlGF result known to clinicians) or nonreveal (result unknown) arms. A ratio cutoff of 38 was used to define low (≤38) and elevated risk (>38) of developing the condition in the subsequent week. The primary end point was hospitalization within 24 hours of the test. Secondary end points were development of preeclampsia and other adverse maternal-fetal outcomes. We recruited 370 women (186 reveal versus 184 nonreveal). Preeclampsia occurred in 85 women (23%). The number of admissions was not significantly different between groups (n=48 nonreveal versus n=60 reveal; P=0.192). The reveal trial arm admitted 100% of the cases that developed preeclampsia within 7 days, whereas the nonreveal admitted 83% (P=0.038). Use of the test yielded a sensitivity of 100% (95% CI, 85.8-100) and a negative predictive value of 100% (95% CI, 97.1-100) compared with a sensitivity of 83.3 (95% CI, 58.6-96.4) and negative predictive value of 97.8 (95% CI, 93.7-99.5) with clinical practice alone. Use of the sFlt-1/PlGF ratio significantly improved clinical precision without changing the admission rate. Clinical Trial Registration- URL: http://www.isrctn.com. Unique identifier: ISRCTN87470468.


Closing the mycetoma knowledge gap.
On 28th May 2016, mycetoma was recognized as a neglected tropical disease by the World Health Organization. This was the result of a 4-year journey starting in February 2013 with a meeting of global mycetoma experts. Knowledge gaps were identified and included the incidence, prevalence, and mapping of mycetoma; the mode of transmission; the development of methods for early diagnosis; and better treatment. In this review, we review the road to recognition, the ISHAM working group meeting in Argentina, and we address the progress made in closing the knowledge gaps since 2013. Progress included adding another 9000 patients to the literature, which allowed us to update the prevalence map on mycetoma. Furthermore, based on molecular phylogeny, species names were corrected and four novel mycetoma causative agents were identified. By mapping mycetoma causative agents an association with Acacia trees was found. For early diagnosis, three different isothermal amplification techniques were developed, and novel antigens were discovered. To develop better treatment strategies for mycetoma patients, in vitro susceptibility tests for the coelomycete agents of black grain mycetoma were developed, and the first randomized clinical trial for eumycetoma started early 2017.


How do drug market changes affect characteristics of injecting initiation and subsequent patterns of drug use? Findings from a cohort of regular heroin and methamphetamine injectors in Melbourne, Australia.
Changes in drug market characteristics have been shown to affect drug use patterns but few studies have examined their impacts on injecting initiation experiences and subsequent patterns of injecting drug use (IDU). We collected data on self-reported injecting initiation experiences and past-month patterns of IDU from 688 regular heroin and methamphetamine injectors in Melbourne, Australia, who initiated injecting across three different drug market periods (prior to the Australian heroin shortage ('high heroin')/immediately following the shortage ('low heroin')/'contemporary' markets (fluctuating heroin and methamphetamine availability)). We used univariable and multivariable logistic regression to examine the relationship between period of injecting initiation and first drug injected, and multinomial logistic regression for the relationship between period of injecting initiation and current injecting patterns. 425 participants (62%) reported initiating injecting in the high heroin period, 146 (21%) in the low heroin period, and 117 (17%) in the contemporary period. Participants who initiated injecting during the low heroin period were twice as likely to initiate injecting using a drug other than heroin (AOR: 1.94, 95% CI: 1.27-2.95). The most common patterns of drug use among study participants in the month preceding interview were polydrug use (44%) and primary heroin use (41%). Injecting initiation period was either non-significantly or weakly associated with current drug use pattern, which was more strongly associated with other socio-demographic and drug use characteristics, particularly self-reported drug of choice. The drug market period in which injecting initiation occurred influenced the first drug injected and influenced some aspects of subsequent drug use. In the context of highly dynamic drug markets in which polydrug use is common there is a need for broad harm reduction and drug treatment services which are flexible and responsive to changing patterns of drug use.


The relation between treatment benefit and underlying risk in meta-analysis.
In meta-analyses of clinical trials comparing a treated group with a control group it has been common to ask whether the treatment benefit varies according to the underlying risk of the patients in the different trials, with the hope of defining which patients would benefit most and which least from medical interventions. The usual analysis used to investigate this issue, however, which uses the observed proportions of events in the control groups of the trials as a measure of the underlying risk, is flawed and produces seriously misleading results. This arises through a bias due to regression to the mean and will be particularly acute in meta-analyses which include some small trials or in which the variability in the true underlying risks across trials is small. Approaches which previously have been thought to be more appropriate are to substitute the average proportion of events in the control and treated groups as the measure of underlying risk or to plot the proportion of events in the treated group against that in the control group (L'Abbé plot). However, these are still subject to bias in most circumstances. Because of the potentially seriously flawed conclusions that can result from such analyses, they should be replaced either by statistically appropriate (but more complex) approaches or, preferably, by analyses which investigate the dependence of the treatment effect on measured baseline characteristics of the patients in each trial.


Mode I and Mode III fractures in intermediate zone of full-thickness porcine temporomandibular joint discs.
The aim of this study was to assess the critical energy required to induce flaw propagation in the temporomandibular joint (TMJ) disc when tensile and shear stresses were applied. J-integrals were measured for Mode I and III fractures because excessive tensile and shear stresses promote disc failure. Single edge notch (Mode I) and trouser tear (Mode III) specimens were constructed with flaws oriented parallel to the predominant anteroposteriorly oriented collagen fibers of the TMJ disc. Disks with and without an impulsive pre-load of 3 N s were studied to compare impact-damaged and healthy tissues. Results demonstrated that impulsive loading stiffened the tissues and significantly increased the Mode I fracture energy (J (IC)) but not Mode III (J (IIIC)) (p < or = 0.05). J (IC) and J (IIIC) values were similar for undamaged tissues, but J (IC) values were 2.3 times higher for impulsively loaded tissues (p < or = 0.05). This suggests that when flaws are introduced through impact, the TMJ disc responds by requiring more energy for tensile flaw extension. This research is a first step towards characterizing the mechanical microenvironment that initiates joint disease. This characterization is essential for successful integration of engineered replacement tissues for damaged TMJs.


Somatization and the vocabulary of everyday bodily experiences and concerns: a community study of adolescents.
To describe the frequency of everyday bodily experiences and health concerns in a general population of adolescents 12 to 16 years of age in Ontario and to explore whether the concept of "somatization," identified from those youths with many of these symptoms, is meaningful and related to other variables. A representative sample of the population was obtained by stratified random sampling. Children with a chronic medical condition were excluded. Parents and their adolescent children filled out a series of questionnaires to measure health concerns, complaints, and more dramatic losses of function. Information was also collected on certain background factors, psychiatric problems, and impairments in adaptive functioning. Parents and youths endorsed the items with the same rank order of frequency, but there was virtually no agreement between parents and youths on the presence or absence of individual somatic symptoms. Users of medical services did not tend to have many more health concerns than others, and there was a weak relationship between the number of health concerns reported by a youth and both impairment in adaptive functioning and psychiatric problems. These data suggest that the concept of somatization has limited general value over and above a relationship with other psychiatric problems.


Consequences of life history switch point plasticity for juvenile morphology and locomotion in the Túngara frog.
Many animals with complex life cycles can cope with environmental uncertainty by altering the timing of life history switch points through plasticity. Pond hydroperiod has important consequences for the fitness of aquatic organisms and many taxa alter the timing of life history switch points in response to habitat desiccation. For example, larval amphibians can metamorphose early to escape drying ponds. Such plasticity may induce variation in size and morphology of juveniles which can result in carry-over effects on jumping performance. To investigate the carry-over effects of metamorphic plasticity to pond drying, we studied the Túngara frog, Physalaemus pustulosus, a tropical anuran that breeds in highly ephemeral habitats. We conducted an outdoor field mesocosm experiment in which we manipulated water depth and desiccation and measured time and size at metamorphosis, tibiofibula length and jumping performance. We also conducted a complimentary laboratory experiment in which we manipulated resources, water depth and desiccation. In the field experiment, metamorphs from dry-down treatments emerged earlier, but at a similar size to metamorphs from constant depth treatments. In the laboratory experiment, metamorphs from the low depth and dry-down treatments emerged earlier and smaller. In both experiments, frogs from dry-down treatments had relatively shorter legs, which negatively impacted their absolute jumping performance. In contrast, reductions in resources delayed and reduced size at metamorphosis, but had no negative effect on jumping performance. To place these results in a broader context, we review past studies on carry-over effects of the larval environment on jumping performance. Reductions in mass and limb length generally resulted in lower jumping performance across juvenile anurans tested to date. Understanding the consequences of plasticity on size, morphology and performance can elucidate the linkages between life stages.


Evaluation of mitochondrial DNA copy number estimation techniques.
Mitochondrial DNA copy number (mtDNA-CN), a measure of the number of mitochondrial genomes per cell, is a minimally invasive proxy measure for mitochondrial function and has been associated with several aging-related diseases. Although quantitative real-time PCR (qPCR) is the current gold standard method for measuring mtDNA-CN, mtDNA-CN can also be measured from genotyping microarray probe intensities and DNA sequencing read counts. To conduct a comprehensive examination on the performance of these methods, we use known mtDNA-CN correlates (age, sex, white blood cell count, Duffy locus genotype, incident cardiovascular disease) to evaluate mtDNA-CN calculated from qPCR, two microarray platforms, as well as whole genome (WGS) and whole exome sequence (WES) data across 1,085 participants from the Atherosclerosis Risk in Communities (ARIC) study and 3,489 participants from the Multi-Ethnic Study of Atherosclerosis (MESA). We observe mtDNA-CN derived from WGS data is significantly more associated with known correlates compared to all other methods (p < 0.001). Additionally, mtDNA-CN measured from WGS is on average more significantly associated with traits by 5.6 orders of magnitude and has effect size estimates 5.8 times more extreme than the current gold standard of qPCR. We further investigated the role of DNA extraction method on mtDNA-CN estimate reproducibility and found mtDNA-CN estimated from cell lysate is significantly less variable than traditional phenol-chloroform-isoamyl alcohol (p = 5.44x10-4) and silica-based column selection (p = 2.82x10-7). In conclusion, we recommend the field moves towards more accurate methods for mtDNA-CN, as well as re-analyze trait associations as more WGS data becomes available from larger initiatives such as TOPMed.


LRRK2 protective haplotype and full sequencing study in REM sleep behavior disorder.
Individuals with rapid eye movement (REM)-sleep behavior disorder (RBD) are likely to progress to synucleinopathies, mainly Parkinson's disease (PD), dementia with Lewy-bodies (DLB) and multiple system atrophy (MSA). The genetics of RBD only partially overlaps with PD and DLB, and the role of LRRK2 variants in risk for RBD is still not clear. The full coding sequence, exon-intron boundaries and 5' and 3' untranslated regions of LRRK2 were sequenced using targeted next-generation sequencing. A total of 350 RBD patients and 869 controls were sequenced, and regression and burden models were used to examine the association between LRRK2 variants and RBD. No pathogenic mutations that are known to cause PD were identified in RBD patients. The p.N551K-p.R1398H-p.K1423K haplotype was associated with a reduced risk for RBD (OR = 0.66, 95% CI 0.44-0.98, p = 0.0055 for the tagging p.N551K substitution). A common variant, p.S1647T, was nominally associated with risk for RBD (OR = 1.28, 95% CI 1.05-1.56, p = 0.029). Burden analysis identified associations with domains and exons that were derived by the variants of the protective haplotype, and no burden of other rare variants was identified. Carriers of the LRRK2 p.N551K-p.R1398H-p.K1423K haplotype have a reduced risk for developing RBD, yet PD-causing mutations probably have minor or no role in RBD. Additional work is needed to confirm these results and to identify the mechanism associated with reduced risk for RBD.


A microfluidic-induced C. elegans sleep state.
An important feature of animal behavior is the ability to switch rapidly between activity states, however, how the brain regulates these spontaneous transitions based on the animal's perceived environment is not well understood. Here we show a C. elegans sleep-like state on a scalable platform that enables simultaneous control of multiple environmental factors including temperature, mechanical stress, and food availability. This brief quiescent state, which we refer to as microfluidic-induced sleep, occurs spontaneously in microfluidic chambers, which allows us to track animal movement and perform whole-brain imaging. With these capabilities, we establish that microfluidic-induced sleep meets the behavioral requirements of C. elegans sleep and depends on multiple factors, such as satiety and temperature. Additionally, we show that C. elegans sleep can be induced through mechanosensory pathways. Together, these results establish a model system for studying how animals process multiple sensory pathways to regulate behavioral states.


Triadic and collaborative play by gorillas in social games with objects.
Interaction with others over objects has until recently been thought lacking in the social play of non-human great apes, in contrast to that of children; even now, only bonobos have been observed to engage in social play involving objects. Human children's triadic interactions with objects involve joint attention, showing and giving, communication that maintains interaction, and sharing of emotions and experiences. We question assertions that chimpanzees, and non-human great apes in general, lack the key characteristics of children's collaborative play. Here, we show that zoo gorillas play games that are both triadic and collaborative. These games were videotaped at the San Francisco Zoo in five different years and involved five different pairings of gorillas. The context was in most cases playfully competitive, involving objects such as balls, bags and leather pieces as foci of joint attention; the ostensible goal in most games was to gain or keep possession of a particular object. In some episodes, roles as possessor or pursuer of an object were exchanged many times; in others, one gorilla retained possession of an object but encouraged pursuit from a partner. Through gaze and gesture, gorillas invited others to: share interest in and attention to objects; share patterns of play; and re-engage after breaks in play. Sometimes, gorillas would assist others in their efforts to engage in collaborative play: older gorillas encouraged younger partners by 'self-handicapping' their own actions. Collaborative games may occur later in the ontogeny of gorillas than in humans, and depend on the challenges and artifacts available in a particular group's habitat.


Individual Differences in the Neural Dynamics of Response Inhibition.
Response inhibition is a widely studied aspect of cognitive control that is particularly interesting because of its applications to clinical populations. Although individual differences are integral to cognitive control, so too is our ability to aggregate information across a group of individuals, so that we can powerfully generalize and characterize the group's behavior. Hence, an examination of response inhibition would ideally involve an accurate estimation of both group- and individual-level effects. Hierarchical Bayesian analyses account for individual differences by simultaneously estimating group and individual factors and compensate for sparse data by pooling information across participants. Hierarchical Bayesian models are thus an ideal tool for studying response inhibition, especially when analyzing neural data. We construct hierarchical Bayesian models of the fMRI neural time series, models assuming hierarchies across conditions, participants, and ROIs. Here, we demonstrate the advantages of our models over a conventional generalized linear model in accurately separating signal from noise. We then apply our models to go/no-go and stop signal data from 11 participants. We find strong evidence for individual differences in neural responses to going, not going, and stopping and in functional connectivity across the two tasks and demonstrate how hierarchical Bayesian models can effectively compensate for these individual differences while providing group-level summarizations. Finally, we validated the reliability of our findings using a larger go/no-go data set consisting of 179 participants. In conclusion, hierarchical Bayesian models not only account for individual differences but allow us to better understand the cognitive dynamics of response inhibition.


Phase II/pharmacodynamic trial of dose-intensive, weekly parenteral hydroxyurea and fluorouracil administered with interferon alfa-2a in patients with refractory malignancies of the gastrointestinal tract.
Combined depletion of pyrimidine and purine DNA precursors has resulted in therapeutic synergism in vitro. The aims of the current study were to test this strategy in patients with refractory tumors and to assess its effects on selected nucleotide pools. A single-institution phase II trial was initiated in patients with advanced carcinomas of the stomach and pancreas. Patients had measurable disease and had no prior chemotherapy except adjuvant fluorouracil (5FU) or gemcitabine. 5FU was administered by CADD + pump at 2.6 g/m(2) intravenously by 24-hour infusion on days 1, 8, 15, 22, 29, and 36. Parenteral hydroxyurea (HU) was administered at 4.3 g/m(2) as a 24-hour infusion concurrently with 5FU. Interferon alfa-2a (IFN-alpha2a) was administered at 9 million units subcutaneously on days 1, 3, and 5 each week. No drug was administered in weeks 7 and 8. Pharmacodynamic studies were performed to assess drug effects on levels of deoxyuridine triphosphate (dUTP) and thymidine triphosphate (TTP) pools in peripheral-blood mononuclear cells (PBMCs) before and 6 hours after treatment using a highly sensitive DNA polymerase assay. There were 53 patients enrolled onto the study (gastric carcinoma, 31; pancreatic carcinoma, 22). The median age was 61 years, with 22% of patients > or = 70 years old. The predominant grade 3 to 4 toxicities were leukopenia (49%), granulocytopenia (55%), and thrombocytopenia (22%). Severe diarrhea occurred in 12%, mucositis in 0%, and vomiting in 10% of patients. Patients > or = 70 years had no greater incidence of toxicities. Among the 30 assessable patients with gastric carcinoma, there were two (7%) complete responders and 11 (37%) partial responders (median duration, 7 months). Among the 21 assessable patients with pancreatic carcinoma, there was one responder. Median survival among all patients with gastric carcinoma was 10 months and 13 months for patients with pancreatic carcinoma. Twenty-three patients had samples studied for levels of dUTP and TTP. There was no change in the levels of TTP before and after treatment. Furthermore, dUTP was detected in only five of 28 samples after treatment with no increase in the dUTP/TTP ratio. Combination therapy with high-dose, weekly infusional HU and 5FU with IFN-alpha2a modulation was well-tolerated with activity in gastric cancer. Patients > or = 70 years tolerated therapy as well as younger patients. This was the first study to correlate levels of TTP and dUTP after treatment with clinical outcome. In PBMCs used as a surrogate tissue, HU abrogated the 5FU-induced increase in dUTP levels without reversing the overall efficacy of the regimen.


ATP-dependent roX RNA remodeling by the helicase maleless enables specific association of MSL proteins.
Dosage compensation in Drosophila involves a global activation of genes on the male X chromosome. The activating complex (MSL-DCC) consists of male-specific-lethal (MSL) proteins and two long, noncoding roX RNAs. The roX RNAs are essential for X-chromosomal targeting, but their contributions to MSL-DCC structure and function are enigmatic. Conceivably, the RNA helicase MLE, itself an MSL subunit, is actively involved in incorporating roX into functional DCC. We determined the secondary structure of roX2 and mapped specific interaction sites for MLE in vitro. Upon addition of ATP, MLE disrupted a functionally important stem loop in roX2. This RNA remodeling enhanced specific ATP-dependent association of MSL2, the core subunit of the MSL-DCC, providing a link between roX and MSL subunits. Probing the conformation of roX in vivo revealed a remodeled stem loop in chromatin-bound roX2. The active remodeling of a stable secondary structure by MLE may constitute a rate-limiting step for MSL-DCC assembly.


Chin surgery V: treatment of the long, nonprojecting chin.
Correction of the long, nonprojecting chin requires both vertical reduction and sagittal augmentation. Wedge excision-based therapy reduces chin height and allows for advancement of the distal segment, but it is associated with at least a 10 percent incidence of mental nerve injury. The authors propose two innovative ways to correct the long, nonprojecting chin. There are two approaches, intraoral and extraoral. With the intraoral approach, following a gingivobuccal incision, a single horizontally oblique osteotomy is made at least 6 mm beneath the mental nerve foramina. The vertically long genial segment is freed and the posterior edge is contoured with a side-cutting burr. The contoured jumping genial segment is secured to the mandible with countersunk screws and contoured in situ to preserve the lower 8 to 10 mm. With the extraoral approach, following a submental incision, the anterior and posterior surfaces of the symphysis are cleared (a double-armed suture is placed through the posterior musculature). A reciprocating saw is used to remove the lower border of the symphysis to reduce the vertical excess. The tagged musculature is resuspended, and a tapered, textured implant is secured to the new symphysis. Aesthetic outcomes using these two techniques were good and there were no complications. Representative patients, operated on by the senior author, illustrate these techniques. Both the intraoral one-cut in situ contoured jumping genioplasty and the extraoral vertical reduction/sagittal augmentation genioplasty reduce excess chin height, control sagittal advancement, provide pogonion projection, and avoid the risks of a standard wedge. Both techniques provide custom projection at the lower pole of the new symphysis.


Risk of relapse after antidepressant discontinuation in anxiety disorders, obsessive-compulsive disorder, and post-traumatic stress disorder: systematic review and meta-analysis of relapse prevention trials.
Objectives To examine the risk of relapse and time to relapse after discontinuation of antidepressants in patients with anxiety disorder who responded to antidepressants, and to explore whether relapse risk is related to type of anxiety disorder, type of antidepressant, mode of discontinuation, duration of treatment and follow-up, comorbidity, and allowance of psychotherapy.Design Systematic review and meta-analyses of relapse prevention trials.Data sources PubMed, Cochrane, Embase, and clinical trial registers (from inception to July 2016).Study selection Eligible studies included patients with anxiety disorder who responded to antidepressants, randomised patients double blind to either continuing antidepressants or switching to placebo, and compared relapse rates or time to relapse.Data extraction Two independent raters selected studies and extracted data. Random effect models were used to estimate odds ratios for relapse, hazard ratios for time to relapse, and relapse prevalence per group. The effect of various categorical and continuous variables was explored with subgroup analyses and meta-regression analyses respectively. Bias was assessed using the Cochrane tool.Results The meta-analysis included 28 studies (n=5233) examining relapse with a maximum follow-up of one year. Across studies, risk of bias was considered low. Discontinuation increased the odds of relapse compared with continuing antidepressants (summary odds ratio 3.11, 95% confidence interval 2.48 to 3.89). Subgroup analyses and meta-regression analyses showed no statistical significance. Time to relapse (n=3002) was shorter when antidepressants were discontinued (summary hazard ratio 3.63, 2.58 to 5.10; n=11 studies). Summary relapse prevalences were 36.4% (30.8% to 42.1%; n=28 studies) for the placebo group and 16.4% (12.6% to 20.1%; n=28 studies) for the antidepressant group, but prevalence varied considerably across studies, most likely owing to differences in the length of follow-up. Dropout was higher in the placebo group (summary odds ratio 1.31, 1.06 to 1.63; n=27 studies).Conclusions Up to one year of follow-up, discontinuation of antidepressant treatment results in higher relapse rates among responders compared with treatment continuation. The lack of evidence after a one year period should not be interpreted as explicit advice to discontinue antidepressants after one year. Given the chronicity of anxiety disorders, treatment should be directed by long term considerations, including relapse prevalence, side effects, and patients' preferences.


Thermal biofeedback for primary Raynaud's phenomenon: a review of the literature.
The clinical presentation of primary Raynaud's phenomenon (RP) derives from various pathogenic triggers. The use of thermal biofeedback (TBF) may be of benefit in reducing the severity and frequency of attacks. This article summarizes the relevant research regarding the pathophysiology of primary RP and mechanism of TBF for RP. Systematic reviews of the efficacy of TBF for RP and treatment guidelines for clinicians are provided. The panel concludes that the level of evidence for TBF efficacy is categorized as Level IV: efficacious. The rationale, based on three randomized controlled trials conducted in independent laboratories, demonstrated "superiority or equivalence" of treatments that include TBF. However, randomly controlled trials (RCT) with positive clinical outcomes tended to be small. A large RCT with negative results did not effectively teach handwarming skills. Procedures for reviewing and rating of the levels of evidence of efficacy of studies was based on the Template for Developing Guidelines for the Evaluation of the Clinical Efficacy of Psychophysiological Interventions developed by the joint task force of the AAPB and the Society for Neuronal Regulation (SNR).


International recommendations for national patient safety incident reporting systems: an expert Delphi consensus-building process.
Patient safety incident reporting systems (PSRS) have been established for over a decade, but uncertainty remains regarding the role that they can and ought to play in quantifying healthcare-related harm and improving care. To establish international, expert consensus on the purpose of PSRS regarding monitoring and learning from incidents and developing recommendations for their future role. After a scoping review of the literature, semi-structured interviews with experts in PSRS were conducted. Based on these findings, a survey-based questionnaire was developed and subsequently completed by a larger expert panel. Using a Delphi approach, consensus was reached regarding the ideal role of PSRSs. Recommendations for best practice were devised. Forty recommendations emerged from the Delphi procedure on the role and use of PSRS. Experts agreed reporting system should not be used as an epidemiological tool to monitor the rate of harm over time or to appraise the relative safety of hospitals. They agreed reporting is a valuable mechanism for identifying organisational safety needs. The benefit of a national system was clear with respect to medication error, device failures, hospital-acquired infections and never events as these problems often require solutions at a national level. Experts recommended training for senior healthcare professionals in incident investigation. Consensus recommendation was for hospitals to take responsibility for creating safety solutions locally that could be shared nationally. We obtained reasonable consensus among experts on aims and specifications of PSRS. This information can be used to reflect on existing and future PSRS, and their role within the wider patient safety landscape. The role of PSRS as instruments for learning needs to be elaborated and developed further internationally.


Peptides as antigens. Importance of orientation.
Factors known to be important in producing protein-reactive peptide antibodies include the accessibility of the region from which the peptide sequence is derived, the hydrophilic-phobic character of the sequence, and the length of the peptide. The data presented here indicate that the orientation of the peptide coupled to a carrier protein also influences the binding pattern of peptide antibodies. An octapeptide, representing a sequence from the alpha chain of the human acetylcholine receptor, was coupled either through an N- or C-terminal cysteine-glycine-glycine linker to a carrier protein and used to immunize rabbits. The resulting antisera reacted at comparable titers to the uncoupled immunizing peptides, but did not crossreact with the identical but opposite-linked peptide. Characterization of the binding to other homologous peptides showed that immunization with the N-terminal-linked peptide induced antibodies reactive specifically with the C-terminal amino acid(s). Immunization with the C-linked peptide resulted in antibodies reactive with a site of the peptide near the C-terminus.


Conscious and unconscious representations of observed actions in the human motor system.
Action observation activates the observer's motor system. These motor resonance responses are automatic and triggered even when the action is only implied in static snapshots. However, it is largely unknown whether an action needs to be consciously perceived to trigger motor resonance. In this study, we used single-pulse TMS to study the facilitation of corticospinal excitability (a measure of motor resonance) during supraliminal and subliminal presentations of implied action images. We used a forward and backward dynamic masking procedure that successfully prevented the conscious perception of prime stimuli depicting a still hand or an implied abduction movement of the index or little finger. The prime was followed by the supraliminal presentation of a still or implied action probe hand. Our results revealed a muscle-specific increase of motor facilitation following observation of the probe hand actions that were consciously perceived as compared with observation of a still hand. Crucially, unconscious perception of prime hand actions presented before probe still hands did not increase motor facilitation as compared with observation of a still hand, suggesting that motor resonance requires perceptual awareness. However, the presentation of a masked prime depicting an action that was incongruent with the probe hand action suppressed motor resonance to the probe action such that comparable motor facilitation was recorded during observation of implied action and still hand probes. This suppression of motor resonance may reflect the processing of action conflicts in areas upstream of the motor cortex and may subserve a basic mechanism for dealing with the multiple and possibly incongruent actions of other individuals.