Target-based vs. phenotypic screenings in Leishmania drug discovery: A marriage of convenience or a dialogue of the deaf?
Drug discovery programs sponsored by public or private initiatives pursue the same ambitious goal: a crushing defeat of major Neglected Tropical Diseases (NTDs) during this decade. Both target-based and target-free screenings have pros and cons when it comes to finding potential small-molecule leads among chemical libraries consisting of myriads of compounds. Within the target-based strategy, crystals of pathogen recombinant-proteins are being used to obtain three-dimensional (3D) structures in silico for the discovery of structure-based inhibitors. On the other hand, genetically modified parasites expressing easily detectable reporters are in the pipeline of target-free (phenotypic) screenings. Furthermore, lead compounds can be scaled up to in vivo preclinical trials using rodent models of infection monitoring parasite loads by means of cutting-edge bioimaging devices. As such, those preferred are fluorescent and bioluminescent readouts due to their reproducibility and rapidity, which reduces the number of animals used in the trials and allows for an earlier stage detection of the infective process as compared with classical methods. In this review, we focus on the current differences between target-based and phenotypic screenings in Leishmania, as an approach that leads to the discovery of new potential drugs against leishmaniasis.


Non-tuberculous Mycobacterial Pseudo-outbreak of an Intestinal Culture Specimen Caused by a Water Tap in an Endoscopy Unit.
Objective Gastrointestinal lesions of non-tuberculous mycobacteria (NTM) are regarded as opportunistic infections. A large number of positive specimens of NTM were identified in an intestinal fluid culture in the endoscopy unit and it was considered to be a pseudo-outbreak. Methods We reviewed the hospital, laboratory, and colonoscopy records of 263 consecutive patients whose intestinal fluids were analyzed for a mycobacterial culture by colonoscopy at St. Marianna University Hospital, between January 2009 and December 2018. The endoscopy reprocessing procedures were reviewed and samples of water used in the endoscopy unit were cultured. Results An intestinal fluid culture of 154 (58.6%) patients tested positive for NTM (M. intracellulare; 125 cases, M. gordonae; 14 cases, M. avium; 4 cases, M. abscessus; 3 cases, and 8 other cases). In 182 cases (69.2%), an intestinal mucosal culture was performed simultaneously with a fluid culture and tested positive for NTM in 2 cases. Next, we examined the endoscopy unit for any possible environmental contamination. NTM were detected in the tap water used to prepare the antifoaming solution in the endoscopy unit. The water faucets in the endoscopy unit were considered to be the source of the contamination of NTMs. Conclusion We observed that a large number of cases tested positive due to contaminated water that had been used in an endoscopy unit, thus leading to a pseudo-outbreak of NTM.


Peripheral neuropathy associated with simvastatin.
Four patients are described who developed sensorimotor neuropathy while being treated with simvastatin and had complete or partial resolution of clinical abnormalities after withdrawal of treatment. In one case onset was within days of commencing treatment, but in two cases symptoms did not develop for two years. The electrophysiological and pathological features of the neuropathy were those of axonal degeneration. Clinical evidence of proximal and distal weakness and muscle fasciculations and persistent abnormalities of sensory conduction after recovery suggest the possibility of toxic damage to anterior horn cells and dorsal root ganglia. Thirty eight other cases with symptoms suggestive of peripheral neuropathy have been reported to the Australian Adverse Drug Reactions Advisory Committee, 22 of whom recovered after cessation of treatment; in five cases there was recurrence after re-exposure to the drug. Simvastatin should be considered among the causes of peripheral neuropathy, and the drug should be withdrawn if patients receiving it develop muscle weakness or sensory disturbances.


Unanswered questions and research priorities to optimise stroke prevention in atrial fibrillation with the new oral anticoagulants.
This review article discusses the following, as yet unanswered, questions and research priorities to optimise patient management and stroke prevention in atrial fibrillation with the new direct oral anticoagulants (NOACs): 1. In patients prescribed a NOAC, can the anticoagulant effects or plasma concentrations of the NOACs be measured rapidly and reliably and, if so, can "cut-off points" between which anticoagulation is therapeutic (i.e. the "therapeutic range") be defined? 2. In patients who are taking a NOAC and bleeding (e.g. intracerebral haemorrhage), can the anticoagulant effects of the direct NOACs be reversed rapidly and, if so, can NOAC-associated bleeding and complications be minimised and patient outcome improved? 3. In patients taking a NOAC who experience an acute ischaemic stroke, to what degree of anticoagulation or plasma concentration of NOAC, if any, can thrombolysis be administered safely and effectively? 4. In patients with a recent cardioembolic ischaemic stroke, what is the optimal time to start (or re-start) anticoagulation with a NOAC (or warfarin)? 5. In anticoagulated patients who experience an intracranial haemorrhage, can anticoagulation with a NOAC be re-started safely and effectively, and if so when? 6. Are the NOACs effective and safe in multimorbid geriatric people (who commonly have atrial fibrillation and are at high risk of stroke but also bleeding)? 7. Can dose-adjusted NOAC therapy augment the established safety and efficacy of fixed-dose unmonitored NOAC therapy? 8. Is there a dose or dosing regimen for each NOAC that is as effective and safe as adjusted-dose warfarin for patients with atrial fibrillation who have mechanical prosthetic heart valves? 9. What is the long-term safety of the NOACs?


Disseminated mycobacteria chelonae infection in a kidney-pancreas transplant recipient: A case report and review of the literature.
A 40-year-old male with a long-standing history of type 1 diabetes with end-stage renal failure underwent combined kidney-pancreas (KP) transplant from a standard criteria donor. Post-operative course was uncomplicated with good primary function of both transplant grafts. Induction was with thymoglobulin and maintenance immunosuppression was with tacrolimus, mycophenolate mofetil and prednisolone. Nine weeks post-transplant, the patient developed dysfunction of both grafts. Panel reactive antibody testing revealed that the patient had developed a de novo donor-specific antibody and considering an antibody-mediated rejection the patient was treated with intravenous pulse methyl prednisone 500 mg ×3 doses, IV immunoglobulin 2 mg/kg in two divided doses, and ATG 7 mg/kg (total dose of 700 mg). In addition, his baseline immunosuppression was increased. Cr decreased to baseline levels, and blood sugars were in the range of 7-8 mmol/L, serum amylase normalized to 63 U/L, and the patient was discharged home. Nine days post-discharge, the patient presented to the hospital with a five-day history of fever, pain, and swelling in the left knee along with subcutaneous, erythematous, tender, nodular lesions in both legs and both arms. Skin biopsy showed Ziehl-Neelsen stain positive rods and biopsy culture and blood culture grew Mycobacteria chelonae. Antimicrobials were switched to azithromycin 500 mg OD, moxifloxacin 400 mg OD, and linezolid 600 mg BID and baseline immunosuppression was reduced to tacrolimus trough target 8-10 ng/mL and MMF to 250 mg BID. The patient gradually improved and was discharged after 28 days in the hospital. Six weeks following the diagnosis of nontuberculous mycobacteria infection, the patient's pancreas graft failed, presumably due to reduction in immuno-suppression and he is now back on insulin treatment. His renal graft continued to function well. Although rapidly growing mycobacterial infections are rare among transplant recipients, it should be suspected among those who have received augmented immunosuppression. Blood cultures and skin biopsy of the lesions are important to establish the diagnosis.


Leiomodin 3 and tropomodulin 4 have overlapping functions during skeletal myofibrillogenesis.
Precise regulation of thin filament length is essential for optimal force generation during muscle contraction. The thin filament capping protein tropomodulin (Tmod) contributes to thin filament length uniformity by regulating elongation and depolymerization at thin filament ends. The leiomodins (Lmod1-3) are structurally related to Tmod1-4 and also localize to actin filament pointed ends, but in vitro biochemical studies indicate that Lmods act instead as robust nucleators. Here, we examined the roles of Tmod4 and Lmod3 during Xenopus skeletal myofibrillogenesis. Loss of Tmod4 or Lmod3 resulted in severe disruption of sarcomere assembly and impaired embryonic movement. Remarkably, when Tmod4-deficient embryos were supplemented with additional Lmod3, and Lmod3-deficient embryos were supplemented with additional Tmod4, sarcomere assembly was rescued and embryonic locomotion improved. These results demonstrate for the first time that appropriate levels of both Tmod4 and Lmod3 are required for embryonic myofibrillogenesis and, unexpectedly, both proteins can function redundantly during in vivo skeletal muscle thin filament assembly. Furthermore, these studies demonstrate the value of Xenopus for the analysis of contractile protein function during de novo myofibril assembly.


Sexual health interventions for young people in state care: a systematic review.
To describe evaluated sexual health interventions for young people in state care and provide an assessment of the quality of and evidence for these interventions. A systematic review of sexual health interventions for young people in state care was conducted. Randomised controlled trials and quasi-experimental designs were eligible, 2051 records were screened, 412 full-text studies retrieved, and 12 publications with low-to-moderate risk of bias included. Due to substantial heterogeneity in study populations, settings, intervention approaches, outcomes and measures, standard summary measures for intervention outcomes was not used. Instead, data were synthesised across studies and presented narratively. Without making recommendations, the result suggests that group-based educational interventions in general increase knowledge, attitudes and behaviour compared with standard care. However, these findings need to be further investigated, with a special emphasis on cultural context and the involvement of young people.


Sharing Rewards Undermines Coordinated Hunting.
Coordinated hunting is widely observed in animals, and sharing rewards is often considered a major incentive for its success. While current theories about the role played by sharing in coordinated hunting are based on correlational evidence, we reveal the causal roles of sharing rewards through computational modeling with a state-of-the-art Multi-agent Reinforcement Learning (MARL) algorithm. We show that counterintuitively, while selfish agents reach robust coordination, sharing rewards undermines coordination. Hunting coordination modeled through sharing rewards (1) suffers from the free-rider problem, (2) plateaus at a small group size, and (3) is not a Nash equilibrium. Moreover, individually rewarded predators outperform predators that share rewards, especially when the hunting is difficult, the group size is large, and the action cost is high. Our results shed new light on the actual importance of prosocial motives for successful coordination in nonhuman animals and suggest that sharing rewards might simply be a byproduct of hunting, instead of a design strategy aimed at facilitating group coordination. This also highlights that current artificial intelligence modeling of human-like coordination in a group setting that assumes rewards sharing as a motivator (e.g., MARL) might not be adequately capturing what is truly necessary for successful coordination.


Polycyclic aromatic hydrocarbon-induced cytotoxicity in cultured rat Sertoli cells involves differential apoptotic response.
Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous and persistent environmental contaminants. Some PAHs are carcinogens and may affect the male reproductive system. Therefore, we exposed cultured rat Sertoli cells to a variety of PAHs to determine possible direct toxic effects on the cells of the seminiferous epithelium. Sertoli cells were chosen because they support germ cell development and maintain spermatogenesis. Sertoli cells were isolated from 19-21-day-old male rats and cultured in medium containing 0.08% dimethylsulfoxide as vehicle or in the presence of a variety of PAHs. In the first set of experiments, cultured Sertoli cells were incubated in the presence of 10(-4) M, 10(-6 )M, 10(-8) M, 10(-12) M, and 10(-16) M fluoranthene (FL) for 24 hr. After 24 hr, FL at 10(4), 10(-6), and 10(-8) M killed significant numbers of Sertoli cells as revealed by cell viability determinations. Sertoli cells cultured in the presence of 10(-6) M and 10(-8) M FL showed morphologic changes. Cell protein levels were decreased and lactate production in the medium increased in a concentration-dependent manner. In addition, Sertoli cells exposed to 10(6) M and 10(-8) M FL exhibited altered F-actin and alpha-tubulin distributions compared with untreated controls. Because FL killed about 62% of cells at 10(-4) M (100 micro g/mL) and 48% of cells at 10(-6) M (1 micro g/mL), increased lactate production about 3-fold at both concentrations, and decreased cell protein by half at 10(-4) M (100 micro g/mL), we decided to use a range of concentrations between 10 and 100 micro g/mL for the second set of experiments using benz[a]anthracene (BaA), benzo[a]pyrene (BaP), or benzo[b]fluoranthene (BbF). After 24 hr, BaA (100 micro g/mL), BaP (50 and 100 micro g/mL), and BbF (100 micro g/mL) significantly increased lactate level in the medium in a concentration-dependent manner. In a third set of experiments, cells were treated in culture uniformly with only 10 micro g/mL FL, BaA, BaP, or BbF for 24 hr. The cytotoxic effects exerted by these PAHs tested resulted in different apoptotic responses as characterized by in situ fluorescence staining. Microscopic analysis of apoptotic cells demonstrated nuclei of reduced size and labeled 3 -OH DNA ends when Sertoli cells had been incubated for 24 hr with 10 micro g BaP or BbF, but not with vehicle, media, FL, or BaA. Thus, our results demonstrate that the toxic effects of BaA and BbF on Sertoli cells are exerted through apoptosis, whereas FL and BaA do not elicit the apoptotic response.


Portraying emotions at their unfolding: a multilayered approach for probing dynamics of neural networks.
Dynamic functional integration of distinct neural systems plays a pivotal role in emotional experience. We introduce a novel approach for studying emotion-related changes in the interactions within and between networks using fMRI. It is based on continuous computation of a network cohesion index (NCI), which is sensitive to both strength and variability of signal correlations between pre-defined regions. The regions encompass three clusters (namely limbic, medial prefrontal cortex (mPFC) and cognitive), each previously was shown to be involved in emotional processing. Two sadness-inducing film excerpts were viewed passively, and comparisons between viewer's rated sadness, parasympathetic, and inter-NCI and intra-NCI were obtained. Limbic intra-NCI was associated with reported sadness in both movies. However, the correlation between the parasympathetic-index, the rated sadness and the limbic-NCI occurred in only one movie, possibly related to a "deactivated" pattern of sadness. In this film, rated sadness intensity also correlated with the mPFC intra-NCI, possibly reflecting temporal correspondence between sadness and sympathy. Further, only for this movie, we found an association between sadness rating and the mPFC-limbic inter-NCI time courses. To the contrary, in the other film in which sadness was reported to commingle with horror and anger, dramatic events coincided with disintegration of these networks. Together, this may point to a difference between the cinematic experiences with regard to inter-network dynamics related to emotional regulation. These findings demonstrate the advantage of a multi-layered dynamic analysis for elucidating the uniqueness of emotional experiences with regard to an unguided processing of continuous and complex stimulation.


Prostaglandin D2 plays an essential role in chronic allergic inflammation of the skin via CRTH2 receptor.
PGD(2) plays roles in allergic inflammation via specific receptors, the PGD receptor designated DP and CRTH2 (chemoattractant receptor homologous molecule expressed on Th2 cells). We generated mutant mice carrying a targeted disruption of the CRTH2 gene to investigate the functional roles of CRTH2 in cutaneous inflammatory responses. CRTH2-deficent mice were fertile and grew normally. Ear-swelling responses induced by hapten-specific IgE were less pronounced in mutant mice, giving 35-55% of the responses of normal mice. Similar results were seen in mice treated with a hemopoietic PGD synthase inhibitor, HQL-79, or a CRTH2 antagonist, ramatroban. The reduction in cutaneous responses was associated with decreased infiltration of lymphocytes, eosinophils, and basophils and decreased production of macrophage-derived chemokine and RANTES at inflammatory sites. In models of chronic contact hypersensitivity induced by repeated hapten application, CRTH2 deficiency resulted in a reduction by approximately half of skin responses and low levels (63% of control) of serum IgE production, although in vivo migration of Langerhans cells and dendritic cells to regional lymph nodes was not impaired in CRTH2-deficient mice. In contrast, delayed-type hypersensitivity to SRBC and irritation dermatitis in mutant mice were the same as in wild-type mice. These findings indicate that the PGD(2)-CRTH2 system plays a significant role in chronic allergic skin inflammation. CRTH2 may represent a novel therapeutic target for treatment of human allergic disorders, including atopic dermatitis.


Genetic variants influencing circulating lipid levels and risk of coronary artery disease.
Genetic studies might provide new insights into the biological mechanisms underlying lipid metabolism and risk of CAD. We therefore conducted a genome-wide association study to identify novel genetic determinants of low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and triglycerides. We combined genome-wide association data from 8 studies, comprising up to 17 723 participants with information on circulating lipid concentrations. We did independent replication studies in up to 37 774 participants from 8 populations and also in a population of Indian Asian descent. We also assessed the association between single-nucleotide polymorphisms (SNPs) at lipid loci and risk of CAD in up to 9 633 cases and 38 684 controls. We identified 4 novel genetic loci that showed reproducible associations with lipids (probability values, 1.6×10(-8) to 3.1×10(-10)). These include a potentially functional SNP in the SLC39A8 gene for HDL-C, an SNP near the MYLIP/GMPR and PPP1R3B genes for LDL-C, and at the AFF1 gene for triglycerides. SNPs showing strong statistical association with 1 or more lipid traits at the CELSR2, APOB, APOE-C1-C4-C2 cluster, LPL, ZNF259-APOA5-A4-C3-A1 cluster and TRIB1 loci were also associated with CAD risk (probability values, 1.1×10(-3) to 1.2×10(-9)). We have identified 4 novel loci associated with circulating lipids. We also show that in addition to those that are largely associated with LDL-C, genetic loci mainly associated with circulating triglycerides and HDL-C are also associated with risk of CAD. These findings potentially provide new insights into the biological mechanisms underlying lipid metabolism and CAD risk.


Protection against Chlamydia trachomatis infection in vitro and modulation of inflammatory response in vivo by membrane-bound glycosaminoglycans.
Glycosaminoglycans (GAG) efficiently inhibit adherence of several strains of Chlamydia trachomatis to cell lines in vitro, but none of the GAG have been able to inhibit infections in vivo. One possible cause for failure of GAG inhibition in vivo is the inability to deliver a sustained concentration of GAG at the mucosal surface. We tested the possibility of enhancing cell protection by increasing the cell-surface concentration of GAG using membrane-anchored GAG (MAG), composed of phosphatidylethanolamine (PE)-linked GAG. These lipid conjugates were originally designed as extracellular phospholipase A2 (PLA2) inhibitors and exhibit a dual effect: the lipid moiety incorporates into the cell membrane, interfering with the action of PLA2 on cell membranes, and the anchored GAG protects the cell membrane from exogenous inflammatory mediators. We tested the ability of MAG to block chlamydia infection in vitro and in vivo. The MAG blocked infection of epithelial cells in vitro when added to the cells at the same time or before infection, but not if added after the bacteria had already invaded the host cells. One of the MAG led to the production of aberrant Chlamydia vacuoles, suggesting it may inhibit intracellular PLA2 associated with development of the vacuole. Although the MAG did not inhibit vaginal infection of mice, they decreased significantly the level of secretion of the inflammatory cytokines TNF-alpha and IFN-gamma but had no effect on secretion of the neutrophil chemokine, macrophage inflammatory protein-2 (MIP-2). Acute and chronic inflammatory cell infiltrates were not altered by MAG treatment. These findings suggest that lipid conjugation of GAG could be used as a novel approach for increasing cell-surface concentrations of GAG. The inconclusive in vivo results might be due to the physical properties of the tested MAG or an insufficient application protocol, and their improvement might provide the desired inhibitory effects.


Plasmonic resonances in optomagnetic metamaterials based on double dot arrays.
We study optical properties of optomagnetic metamaterials produced by regular arrays of double gold dots (nanopillars). Using combined data of spectroscopic ellipsometry, transmission and reflection measurements, we identify localized plasmon resonances of a nanopillar pair and measure their dependences on dot sizes. We formulate the necessary condition at which an effective field theory can be applied to describe optical properties of a composite medium and employ interferometry to measure phase shifts for our samples. A negative phase shift for transmitted green light coupled to an antisymmetric magnetic mode of a double-dot array is observed.


First Report of Anthracnose of Tricyrtis macropoda Caused by Colletotrichum gloeosporioides in Korea.
Tricyrtis macropoda Miq. (syn. T. dilatata Nakai), known as speckled toadlily, is a perennial herb native to China, Japan, and Korea. The plant has been highly praised for its beautiful flowers and rare populations in natural habitats. In September 2006, several dozen plants were heavily damaged by leaf spots and blight in cultivated plantings in the city of Pocheon, Korea. The infections with the same symptoms were repeated every year. In July 2011, the same symptoms were found on T. macropoda in the cities of Gapyeong and Osan, Korea. The leaf lesions began as small, water-soaked, pale greenish to grayish spots, which enlarged to form concentric rings and ultimately coalesced. A number of blackish acervuli were formed in the lesions. Acervuli were mostly epiphyllous, circular to ellipsoid, and 40 to 200 μm in diameter. Setae were two- to three-septate, dark brown at the base, paler upwards, acicular, and up to 100 μm long. Conidia (n = 30) were long obclavate to oblong-elliptical, sometimes fusiform-elliptical, guttulate, hyaline, and 12 to 20 × 4 to 6.5 μm (mean 15.4 × 5.2 μm). These morphological characteristics of the fungus were consistent with the description of Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. (2). Voucher specimens (n = 7) were deposited in the Korea University herbarium (KUS). Two isolates, KACC46374 (ex KUS-F25916) and KACC46405 (ex KUS-F26063), were deposited in the Korean Agricultural Culture Collection. Fungal DNA was extracted and the complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The resulting sequences of 549 bp were deposited in Genbank (Accession Nos. JQ619480 and JQ619481). They showed 100% similarity with a sequence of C. gloeosporioides (EU32619). Isolate KACC46374 was used in a pathogenicity test. Inoculum was prepared by harvesting conidia from 3-week-old cultures on potato dextrose agar. A conidial suspension (2 × 106 conidia/ml) was sprayed onto 15 leaves of three plants. Three noninoculated plants served as controls. Plants were covered with plastic bags to maintain 100% relative humidity for 24 h and then kept in a greenhouse (22 to 28°C and 70 to 80% RH). After 5 days, typical leaf spot symptoms, identical to the ones observed in the field, started to develop on the leaves of inoculated plants. No symptoms were observed on control plants. C. gloeosporioides was reisolated from the lesions of inoculated plants, thus fulfilling Koch's postulates. An anthracnose associated with C. tricyrtii (Teng) Teng was recorded on T. formosana and T. latifolia in China (3) and on T. formosana in Taiwan (1), respectively, without etiological studies. The morphological features of C. tricyrtii are within the variation of C. gloeosporioides (2). To our knowledge, this is the first report of anthracnose of T. macropoda. This report has significance to indigenous plant resource conservation managers and scientists because T. macropoda has been listed as one of the 126 "Rare and Endangered Plants" by the Korea Forest Service since 1991. References: (1) K. Sawada. Rep. Dept. Agric. Gov. Res. Inst. Formosa 87: 1, 1944. (2) B. C. Sutton. Pages 1-27 in: Colletotrichum Biology, Pathology and Control. J. A. Bailey and M. J. Jeger, eds. CAB International, Wallingford, U.K. 1992. (3) S. C. Teng. Contrib. Biol. Lab. Sci. Soc. China 8:36, 1932.


Organisation and informational content of the Theileria parva genome.
When compared with other Apicomplexan organisms, Theileria parva has an exceptionally small, 10-12 Mbp, genome. There are only 4 chromosomes, each in the Mbp range, and a complete physical map, based on SfiI linking data, is available for each one. A number of genes and cDNAs have been mapped to specific SfiI fragments. Telomeres consist of the simple repeat sequence typical of chromosomal ends but sub-telomeric homologies do not extend beyond 5 kbp. The only dispersed repetitive sequences identified to date are minisatellites, but these are found on a subset of SfiI fragments. There are clusters of distinct multicopy sequences which contain ORFs. However, the majority of parasite protein coding genes are present in a single copy. They have a compact structure, exhibit a bias in codon usage and non-translated regions are small. Introns, if present, are unusually short. Overall, the genome contains remarkably little repetitive, non-coding DNA. The parasite mitochondrial DNA is linear in structure, has a limited protein coding capacity and fragmented rRNA genes and its telomeres contain inverted repeat sequences.


Current status of molecularly targeted therapy for hepatocellular carcinoma: basic science.
Conventional systemic chemotherapy has been developed with so-called anti-cancer agents, essentially screened for cytotoxicity to cultured cancer cells. However, in patients with hepatocellular carcinoma (HCC), the role of chemotherapy is quite limited because most anti-cancer agents are ineffective and relatively toxic to HCC patients with chronic liver diseases. On the other hand, accumulated understanding of the molecular mechanisms regulating cancer progression has led to novel development of molecularly targeted therapies with cytostatic agents. Recently, a phase III clinical trial revealed a multi-kinase inhibitor, Sorafenib, as the first agent leading to improved overall survival of patients with advanced HCC. A new era of HCC treatment has arrived, based on identification of deranged signaling pathways of cancer cells or their microenvironment. This review summarizes the molecular hallmarks of HCC with a focus on angiogenesis, growth signals, and mitotic stress, and a novel concept "synthetic lethality" for the targeted therapy strategy.


Rpd3p relocation mediates a transcriptional response to rapamycin in yeast.
Treating yeast cells with rapamycin, a small molecule that inhibits the TOR proteins, leads to the repression of many genes. Consistent with prior studies, we find that RPD3, which encodes a histone deacetylase (HDAC), is required for repression upon rapamycin treatment. To elucidate the mechanism underlying RPD3-mediated repression, we screened all promoters in yeast for occupancy by Rpd3p before and after treatment with rapamycin. We find that Rpd3p binds to the promoters of rapamycin-repressible genes only following treatment. These data conflict with a previously proposed model suggesting that Rpd3p is constitutively bound to rapamycin-repressible genes and becomes active only after a stimulus such as treatment with rapamycin. Rather, the comprehensive analysis presented here strongly supports a model in which recruitment of Rpd3p to gene promoters is a regulated step in the control of gene repression.


The Gambling Craving Scale: Psychometric validation and behavioral outcomes.
Although craving is an important feature of problem gambling, there is a paucity of research investigating craving to gamble. A major stumbling block for craving research in gambling has been the lack of a methodologically sound, multidimensional measure of gambling-related craving. This article reports the development of the Gambling Craving Scale (GACS). In Study 1 (N = 220), a factor analysis revealed the emergence of a 9-item scale with 3 factors: Anticipation, Desire, and Relief. An important finding was that the GACS predicted problem gambling severity, depression, and positive and negative affect. In Study 2 (N = 145), the factor structure of the GACS was confirmed using a community sample of gamblers. In Study 3 (N = 46), GACS scores significantly predicted persistence at play on a virtual slot machine in the face of continued loss. Specifically, the more participants craved to gamble, the longer they engaged in play. The implications of craving for the development and maintenance of problem gambling severity are discussed.


Hospital nurse staffing models and patient and staff-related outcomes.
Nurse staffing interventions have been introduced across countries in recent years in response to changing patient requirements, developments in patient care, and shortages of qualified nursing staff. These include changes in skill mix, grade mix or qualification mix, staffing levels, nursing shifts or nurses' work patterns. Nurse staffing has been closely linked to patient outcomes, organisational outcomes such as costs, and staff-related outcomes. Our aim was to explore the effect of hospital nurse staffing models on patient and staff-related outcomes. We searched the following databases from inception through to May 2009: Cochrane/EPOC resources (DARE, CENTRAL, the EPOC Specialised Register), PubMed, EMBASE, CINAHL Plus, CAB Health, Virginia Henderson International Nursing Library, the Joanna Briggs Institute database, the British Library, international theses databases, as well as generic search engines. Randomised control trials, controlled clinical trials, controlled before and after studies and interrupted time series analyses of interventions relating to hospital nurse staffing models. Participants were patients and nursing staff working in hospital settings. We included any objective measure of patient or staff-related outcome. Seven reviewers working in pairs independently extracted data from each potentially relevant study and assessed risk of bias. We identified 6,202 studies that were potentially relevant to our review. Following detailed examination of each study, we included 15 studies in the review. Despite the number of studies conducted on this topic, the quality of evidence overall was very limited. We found no evidence that the addition of specialist nurses to nursing staff reduces patient death rates, attendance at the emergency department, or readmission rates, but it is likely to result in shorter patient hospital stays, and reductions in pressure ulcers. The evidence in relation to the impact of replacing Registered Nurses with unqualified nursing assistants on patient outcomes is very limited. However, it is suggested that specialist support staff, such as dietary assistants, may have an important impact on patient outcomes. Self-scheduling and primary nursing may reduce staff turnover. The introduction of team midwifery (versus standard care) may reduce medical procedures in labour and result in a shorter length of stay without compromising maternal or perinatal safety. We found no eligible studies of educational interventions, grade mix interventions, or staffing levels and therefore we are unable to draw conclusions in relation to these interventions. The findings suggest interventions relating to hospital nurse staffing models may improve some patient outcomes, particularly the addition of specialist nursing and specialist support roles to the nursing workforce. Interventions relating to hospital nurse staffing models may also improve staff-related outcomes, particularly the introduction of primary nursing and self-scheduling. However, these findings should be treated with extreme caution due to the limited evidence available from the research conducted to date.


Feature-Based Attention and Feature-Based Expectation.
Foreknowledge of target stimulus features improves visual search performance as a result of 'feature-based attention' (FBA). Recent studies have reported that 'feature-based expectation' (FBE) also heightens decision sensitivity. Superficially, it appears that the latter work has simply rediscovered (and relabeled) the effects of FBA. However, this is not the case. Here we explain why.


Serum holotranscobalamine, vitamin B12, folic acid and homocysteine levels in alopecia areata patients.
Alopecia areata has been associated with many autoimmune diseases. There is a common belief that the prevalence of pernicious anemia is increased in patients with alopecia areata. In this study, we sought to investigate vitamin B12, folate and homocysteine metabolism in alopecia areata. We measured holotranscobalamine (holoTC), vitamin B12, folate and homocysteine levels in 75 patients with alopecia areata and 54 controls. We did not find any significant differences in these parameters between these groups. We think that alopecia areata may not be associated with vitamin B12 deficiency. The co-occurrence of pernicious anemia and alopecia areata in rare autoimmune syndromes, may not justify routine measurements of these parameters in alopecia areata patients.


The segment polarity gene armadillo interacts with the wingless signaling pathway in both embryonic and adult pattern formation.
The segment polarity genes of Drosophila were initially defined as genes required for pattern formation within each embryonic segment. Some of these genes also function to establish the pattern of the adult cuticle. We have examined the role of the armadillo (arm) gene in this latter process. We confirmed and extended earlier findings that arm and the segment polarity gene wingless are very similar in their effects on embryonic development. We next discuss the role of arm in pattern formation in the imaginal discs, as determined by using a pupal lethal allele, by analyzing clones of arm mutant tissue in imaginal discs, and by using a transposon carrying arm to produce adults with a reduced level of arm. Together, these experiments established that arm is required for the development of all imaginal discs. The requirement for arm varies along the dorsal-ventral and proximal-distal axes. Cells that require the highest levels of arm are those that express the wingless gene. Further, animals with reduced arm levels have phenotypes that resemble those of weak alleles of wingless. We present a description of the patterns of arm protein accumulation in imaginal discs. Finally, we discuss the implications of these results for the role of arm and wingless in pattern formation.


Genetic interaction of Lobe with its modifiers in dorsoventral patterning and growth of the Drosophila eye.
Dorsoventral (DV) patterning is essential for growth of the Drosophila eye. Recent studies suggest that ventral is the default state of the early eye, which depends on Lobe (L) function, and that the dorsal fate is established later by the expression of the dorsal selector gene pannier (pnr). However, the mechanisms of regulatory interactions between L and dorsal genes are not well understood. For studying the mechanisms of DV patterning in the early eye disc, we performed a dominant modifier screen to identify additional genes that interact with L. The criterion of the dominant interaction was either enhancement or suppression of the L ventral eye loss phenotype. We identified 48 modifiers that correspond to 16 genes, which include fringe (fng), a gene involved in ventral eye patterning, and members of both Hedgehog (Hh) and Decapentaplegic (Dpp) signaling pathways, which promote L function in the ventral eye. Interestingly, 29% of the modifiers (6 enhancers and 9 suppressors) identified either are known to interact genetically with pnr or are members of the Wingless (Wg) pathway, which acts downstream from pnr. The detailed analysis of genetic interactions revealed that pnr and L mutually antagonize each other during second instar of larval development to restrict their functional domains in the eye. This time window coincides with the emergence of pnr expression in the eye. Our results suggest that L function is regulated by multiple signaling pathways and that the mutual antagonism between L and dorsal genes is crucial for balanced eye growth.


The benefits and challenges of using computer-assisted symptom assessments in oncology clinics: results of a qualitative assessment.
Developed for clinical use in oncology settings, the Patient Assessment, Care & Education (PACE) System is a computer technology tool designed to address the under-identification and treatment of chemotherapy-related symptoms. This system includes general core questions together with the Patient Care Monitor (PCM), a validated questionnaire that assesses patient-reported problems, six symptom burden indices, and one global quality of life index. The system automatically scores the PCM and generates a written report. The objective of this study was to assess the manner in which clinicians use this system and identify the benefits and challenges that oncology clinics may face when adopting this system. The study was part of a larger evaluation of the system that included standardized surveys and chart review. Sixteen providers (physicians, nurses, and physician assistants) at 13 community oncology clinics participated in a 30-minute interview. Responses were coded according to common phrases or concepts. Clinicians indicated that they use the system mainly for symptom assessment or review of systems. The most common benefits identified included the improved ability to identify under-reported symptoms, enhanced communication with patients; increased efficiency; and its ability to highlight patients' most bothersome symptoms. Challenges included patient burden from the frequent need to answer the questionnaires, issues with the wording and formatting of the screening questionnaire, and technical difficulties. In sum, these interviews suggest that electronic symptom assessments offer potential advantages in terms improving the integration of routine assessment of patients' symptoms and health-related quality of life into the daily flow of an oncology clinic. The approach should receive additional research and development attention.


The Effects of Pycnogenol® as Add-on Drug to Metformin Therapy in Diabetic Rats.
The progression of diabetes mellitus leads in time to the development of serious cardiovascular complications. Pycnogenol® (PYC) belongs to strong antioxidants that may interfere with different pathways playing an important role in diseases associated with oxidative stress. Metformin (MET), commonly used antidiabetic drug, has cardio-protective effects via activation of AMP kinase (AMPK). In our study, we examined the effects of PYC as add-on drug to metformin therapy in streptozotocin (STZ)-induced diabetic rats. Our results revealed that both used agents, PYC and MET, showed improvement of blood glucose levels, vascular reactivity, left ventricular hypertrophy, expression of AMPK, glucose transporter 4 (GLUT4) and calcium/calmodulin-dependent protein kinase II (CaMKII) in left ventricle of the hearts. However, the combination of these interventions has failed to possess higher efficacy. Copyright © 2016 John Wiley & Sons, Ltd.


A practice pathway for the identification, evaluation, and management of insomnia in children and adolescents with autism spectrum disorders.
This report describes the development of a practice pathway for the identification, evaluation, and management of insomnia in children and adolescents who have autism spectrum disorders (ASDs). The Sleep Committee of the Autism Treatment Network (ATN) developed a practice pathway, based on expert consensus, to capture best practices for an overarching approach to insomnia by a general pediatrician, primary care provider, or autism medical specialist, including identification, evaluation, and management. A field test at 4 ATN sites was used to evaluate the pathway. In addition, a systematic literature review and grading of evidence provided data regarding treatments of insomnia in children who have neurodevelopmental disabilities. The literature review revealed that current treatments for insomnia in children who have ASD show promise for behavioral/educational interventions and melatonin trials. However, there is a paucity of evidence, supporting the need for additional research. Consensus among the ATN sleep medicine committee experts included: (1) all children who have ASD should be screened for insomnia; (2) screening should be done for potential contributing factors, including other medical problems; (3) the need for therapeutic intervention should be determined; (4) therapeutic interventions should begin with parent education in the use of behavioral approaches as a first-line approach; (5) pharmacologic therapy may be indicated in certain situations; and (6) there should be follow-up after any intervention to evaluate effectiveness and tolerance of the therapy. Field testing of the practice pathway by autism medical specialists allowed for refinement of the practice pathway. The insomnia practice pathway may help health care providers to identify and manage insomnia symptoms in children and adolescents who have ASD. It may also provide a framework to evaluate the impact of contributing factors on insomnia and to test the effectiveness of nonpharmacologic and pharmacologic treatment strategies for the nighttime symptoms and daytime functioning and quality of life in ASD.


Influenza A Viruses in Whistling Ducks (Subfamily Dendrocygninae).
As compared to other Anseriformes, data related to influenza A virus (IAV) detection and isolation, and IAV antibody detection in whistling ducks (Dendrocygna spp. and Thalassornis leuconotus; subfamily Dendrocygninae) are limited. To better evaluate the potential role of whistling ducks in the epidemiology of IAV, we (1) conducted surveillance for IAV from black-bellied whistling ducks (BBWD, Dendrocygnaautumnalis) sampled in coastal Louisiana, USA, during February 2018 and 2019, and (2) reviewed the published literature and Influenza Resource Database (IRD) that reported results of IAV surveillance of whistling ducks. In the prospective study, from 166 BBWD sampled, one H10N7 IAV was isolated (0.6% prevalence), and overall blocking enzyme-linked immunosorbent assay (bELISA) antibody seroprevalence was 10%. The literature review included publications and data in the IRD from 1984 to 2020 that reported results from nearly 5000 collected samples. For any given collection, the IAV isolation rate never exceeded 5.5%, and seroprevalence estimates ranged from 0 to 42%. Results from our prospective study in Louisiana are consistent with this historic literature; however, although all data consistently demonstrated a low prevalence of infection, the potential role of this species in the epidemiology of IAV should not be totally discounted. In sum, whistling ducks can be infected with IAV, they represent important species on many areas where waterfowl winter, and their distribution across the globe appears to be changing.


Guanidinoacetate methyltransferase (GAMT) deficiency: outcomes in 48 individuals and recommendations for diagnosis, treatment and monitoring.
We collected data on 48 patients from 38 families with guanidinoacetate methyltransferase (GAMT) deficiency. Global developmental delay/intellectual disability (DD/ID) with speech/language delay and behavioral problems as the most affected domains was present in 44 participants, with additional epilepsy present in 35 and movement disorder in 13. Treatment regimens included various combinations/dosages of creatine-monohydrate, l-ornithine, sodium benzoate and protein/arginine restricted diets. The median age at treatment initiation was 25.5 and 39 months in patients with mild and moderate DD/ID, respectively, and 11 years in patients with severe DD/ID. Increase of cerebral creatine and decrease of plasma/CSF guanidinoacetate levels were achieved by supplementation with creatine-monohydrate combined with high dosages of l-ornithine and/or an arginine-restricted diet (250 mg/kg/d l-arginine). Therapy was associated with improvement or stabilization of symptoms in all of the symptomatic cases. The 4 patients treated younger than 9 months had normal or almost normal developmental outcomes. One with inconsistent compliance had a borderline IQ at age 8.6 years. An observational GAMT database will be essential to identify the best treatment to reduce plasma guanidinoacetate levels and improve long-term outcomes.


Shewanella irciniae sp. nov., a novel member of the family Shewanellaceae, isolated from the marine sponge Ircinia dendroides in the Bay of Villefranche, Mediterranean Sea.
Strain UST040317-058(T), comprising non-pigmented, rod-shaped, facultatively anaerobic, Gram-negative cells that are motile by means of single polar flagella, was isolated from the surface of a marine sponge (Ircinia dendroides) collected from the Mediterranean Sea. Comparative 16S rRNA gene sequence-based phylogenetic analysis placed the strain in a separate cluster with the recognized bacterium Shewanella algae IAM 14159(T), with which it showed a sequence similarity of 95.0 %. The sequence similarity between strain UST040317-058(T) and its other (six) closest relatives ranged from 91.6 to 93.8 %. Strain UST040317-058(T) showed oxidase, catalase and gelatinase activities. The typical respiratory quinones for shewanellas, menaquinone MK-7 and ubiquinones Q-7 and Q-8, were also detected. The predominant fatty acids in strain UST040317-058(T) were i15 : 0, 16 : 0, 17 : 1omega8c and summed feature 3 (comprising i15 : 0 2-OH and/or 16 : 1omega7c), altogether representing 56.9 % of the total. The DNA G+C content was 39.9 mol%. The strain could be differentiated from other Shewanella species by its inability to reduce nitrate or produce H(2)S and by 10-22 additional phenotypic characteristics. On the basis of the phylogenetic and phenotypic data presented in this study, strain UST040317-058(T) represents a novel species in the genus Shewanella, for which the name Shewanella irciniae sp. nov. is proposed. The type strain is UST040317-058(T) (=JCM 13528(T)=NRRL B-41466(T)).


Survey results of pain treatments in adults with cerebral palsy.
The aims of this study were to identify the types and frequencies of pain treatments used by individuals with cerebral palsy, examine the perceived effectiveness of these treatments, and identify the types of healthcare providers that were accessed for pain-related services. A cross-sectional survey design was used. A total of 83 adults (mean [SD] age, 40.3 [13.6] yrs) with cerebral palsy indicated their pain location and intensity during the past 3 mos. Next, they indicated their use of 24 different pain treatments and the effectiveness of each. Finally, participants indicated the frequency of pain-related healthcare visits to specific providers over the past 6 mos. Of the participants, 63% reported experiencing chronic pain and rated their pain intensity over the past week as 5.1 of 10, on average. The most common pain locations were the lower back, hips, and legs. Physical interventions (e.g., physical therapy, strengthening) were the most common pain treatments reportedly used and were rated as moderately effective. Many other treatments were also used, and participants sought pain-related care from a variety of providers. Although participants reportedly accessed pain care from a variety of providers and perceived that several types of treatments were effective, many of the treatments rated as effective were rarely used or provided. Future research using clinical trial methods would further elucidate the specific pain treatments that are most beneficial for adults with cerebral palsy.


Working Group on Treatment Options for the Maintenance of Marginal Bone Around Endosseous Oral Implants, Stockholm, Sweden, 8 and 9 September 2011. Methodology.
To reach through intense interactions among a group of renowned experts a better view of the treatment options available for marginal bone maintenance and eventual inflammation and/or loss around oral implants. The working group was intended to generate guidelines useful for th clinical community at large. FORMAT: Two days of interactive presentations and discussions on the basis of review papers delivered beforehand by the participants. Each of the participants was asked to focus on a particular aspect of the global theme--maintenance of marginal bone around oral implants and treatment options while keeping a global view for further discussion. From the prepared reviews only the conclusions were presented at the meeting as an introduction for the discussions. The reviews were distributed among all participants beforehand and, if needed, the other participants could draw the attention of the author on missing aspects and references prior to the meeting. A general consensus was formulated. The consensus was amended during the two days of the meeting until a unanimous opinion was reached. Minority statements could be considered. The review papers were not required to be systematic reviews (time and means were lacking) but they were to be valuable critical analyses considering the internationally recognised expertise of the authors.


Comparative genomics of Brassica oleracea and Arabidopsis thaliana reveal gene loss, fragmentation, and dispersal after polyploidy.
We sequenced 2.2 Mb representing triplicated genome segments of Brassica oleracea, which are each paralogous with one another and homologous with a segmentally duplicated region of the Arabidopsis thaliana genome. Sequence annotation identified 177 conserved collinear genes in the B. oleracea genome segments. Analysis of synonymous base substitution rates indicated that the triplicated Brassica genome segments diverged from a common ancestor soon after divergence of the Arabidopsis and Brassica lineages. This conclusion was corroborated by phylogenetic analysis of protein families. Using A. thaliana as an outgroup, 35% of the genes inferred to be present when genome triplication occurred in the Brassica lineage have been lost, most likely via a deletion mechanism, in an interspersed pattern. Genes encoding proteins involved in signal transduction or transcription were not found to be significantly more extensively retained than those encoding proteins classified with other functions, but putative proteins predicted in the A. thaliana genome were underrepresented in B. oleracea. We identified one example of gene loss from the Arabidopsis lineage. We found evidence for the frequent insertion of gene fragments of nuclear genomic origin and identified four apparently intact genes in noncollinear positions in the B. oleracea and A. thaliana genomes.


Is grip strength a predictor for total muscle strength in healthy children, adolescents, and young adults?
The primary purpose of this study was to examine whether grip strength is related to total muscle strength in children, adolescents, and young adults. The second purpose was to provide reference charts for grip strength, which could be used in the clinical and research setting. This cross-sectional study was performed at primary and secondary schools and the University of Applied Sciences. Three hundred and eighty-four healthy Dutch children, adolescents, and young adults at the age of 8 to 20 years participated. Isometric muscle strength was measured with a handheld dynamometer of four muscle groups (shoulder abductors, grip strength, hip flexors, and ankle dorsiflexors). Total muscle strength was a summing up of shoulder abductors, hip flexors, and ankle dorsiflexors. All physical therapists participated in a reliability study. The study was started when intratester and intertester reliability was high (Pearson correlation coefficient >0.8). Grip strength was strongly correlated with total muscle strength, with correlation coefficients between 0.736 and 0.890 (p < 0.01). However, the correlation was weaker when controlled for weight (0.485-0.564, p < 0.01). Grip strength is related to total muscle strength. This indicates, in the clinical setting, that grip strength can be used as a tool to have a rapid indication of someone's general muscle strength. The developed reference charts are suitable for evaluating muscle strength in children, adolescents, and young adults in clinical and research settings.


Rogue waves in injected semiconductor lasers with current modulation: role of the modulation phase.
Semiconductor lasers with continuous-wave optical injection display a rich variety of behaviors, including stable locking, periodic or chaotic oscillations, excitable pulses, etc. Within the chaotic regime it has been shown that the laser intensity can display extreme pulses, which have been identified as optical rogue waves (RWs), and it has also been shown that such extreme pulses can be completely suppressed via direct modulation of the laser current, with appropriated modulation amplitude and frequency. Here we perform a numerical analysis of the RW statistics and show that, when RWs are not suppressed by current modulation, their probability of occurrence strongly depends on the phase of the modulation. If the modulation is slow (the modulation frequency, fmod, is below the relaxation oscillation frequency, fro), the RWs occur within a well-defined interval of values of the modulation phase, i.e., there is a "safe" window of phases where no RWs occur. The most extreme RWs occur for modulation phases that are at the boundary of the safe window. When the modulation is fast (fmod > fro), there is no safe phase window; however, the RWs are likely to occur at particular values of the modulation phase. Our findings are of interest for the study of RWs in other systems, where a similar response to external forcing could be observed, and we hope that they will motivate experimental investigations to further elucidate the role of the modulation phase in the likelihood of the occurrence of RWs.


Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors.
Synthetic receptors provide a powerful experimental tool for generation of designer cells capable of monitoring the environment, sensing specific input signals, and executing diverse custom response programs. To advance the promise of cellular engineering, we have developed a class of chimeric receptors that integrate a highly programmable and portable nuclease-deficient CRISPR/Cas9 (dCas9) signal transduction module. We demonstrate that the core dCas9 synthetic receptor (dCas9-synR) architecture can be readily adapted to various classes of native ectodomain scaffolds, linking their natural inputs with orthogonal output functions. Importantly, these receptors achieved stringent OFF/ON state transition characteristics, showed agonist-mediated dose-dependent activation, and could be programmed to couple specific disease markers with diverse, therapeutically relevant multi-gene expression circuits. The modular dCas9-synR platform developed here provides a generalizable blueprint for designing next generations of synthetic receptors, which will enable the implementation of highly complex combinatorial functions in cellular engineering.


Epilepsy treatment patterns among patients with tuberous sclerosis complex.
Tuberous sclerosis complex (TSC) is a rare congenital disorder often associated with epilepsy. However, real-world treatment patterns for epilepsy in patients with TSC are not yet well categorized. This study included patients with TSC and epilepsy from fifteen clinics in the United States and one in Belgium who were enrolled in the TSC Natural History Database (2006-2014). Patient demographics and epilepsy treatment patterns, including the use of anti-epileptic drugs (AEDs), epilepsy surgeries, and dietary therapies were assessed. Of the 1328 patients with TSC in the database, 1110 (83.6%) were diagnosed with epilepsy. The median age of epilepsy diagnosis was 0.7 years. Of those who received treatment for epilepsy (92.3%), 99.5% were prescribed AEDs, 25.3% underwent surgery, 7.9% were prescribed special diets, and 1% were prescribed mammalian target of rapamycin (mTOR) inhibitors. Of the patients receiving AEDs, over half (64.5%) used ≥3 different AEDs, and 22.5% underwent surgical treatment following AED initiation. Of the patients who underwent surgery, 35.1% had subsequent surgery. The use of multiple AEDs and surgical interventions may indicate a need for new therapies to reduce the treatment burden among patients with TSC and epilepsy.


Mature T cells generated from single thymic clones are phenotypically and functionally heterogeneous.
A limiting dilution system for cloning thymic CFU (CFUt) from murine bone marrow has been critically evaluated to test the clonal origin of the thymic colonies. Simultaneous limiting dilution transfer of three populations of bone marrow, each expressing a unique allelic cell surface determinant, resulted in independent segregation of donor-derived thymocyte populations within groups of recipient mice. Statistical analysis of the data allowed an estimate of 1 CFUt/3.3 x 10(4) i.v. transferred bone marrow cells. A pulse-chase experiment was utilized to establish whether CFUt seed directly to the thymus, or whether thymic seeding is secondary to extra-thymic engraftment. The results supported the conclusion that bone marrow CFUt utilize a specific interaction with thymic blood vessel endothelial cells to recognize and enter the thymus, and that this seeding occurs within 4 h of i.v. infusion. A kinetic analysis of emigration of the CFUt progeny into the peripheral blood revealed that, in most cases, an early wave of predominantly CD4+ CD8- lymphocytes emerges from the thymus approximately 4 wk after radiation and reconstitution. In a few cases, the first progeny of CFUt to emerge from the thymus were predominantly CD4- CD8+. Commitment of CFUt to TCR beta-chain rearrangements was assessed by quantitating expression of the V beta 8 family of TCR V region genes. Although some clones expressed a significantly higher or lower percentage of V beta 8+ cells, these differences were not stable with time. Thus, CFUt do not undergo absolute commitment to cell surface phenotype of TCR rearrangement, as reflected by the phenotypes of their progeny. Clones of mature peripheral progeny of CFUt could be expanded in culture in the presence of mitogen and growth factors; approximately 30 to 50% of proliferating clones could mediate cytotoxicity in a lectin-dependent assay, further indicating that CFUt are not absolutely committed to a particular T cell function.


Selective instability: maternal effort and the evolution of gene activation and deactivation rates.
We previously used simulations of gene expression to demonstrate that rapid activation and deactivation rates stabilized outcomes in stochastic systems. We hypothesized that transient single allele inactivation of an autosomal gene during gametogenesis or very early embryogenesis could have a selective advantage if it permits the functional sampling of each allele and precludes committing maternal effort to an embryo with a deleterious mutation. To test this hypothesis, we simulated the evolution of gene expression activation and deactivation rates and imposed two different selective pressures on the populations: (a). late selection against individuals that cannot maintain a threshold level of gene product that occurs after the investment of maternal effort (i.e., after birth); or (b). early selection: in addition to late selection, maintenance of the gene product above a threshold level was necessary for early development prior to commitment of maternal effort. We found that the opportunity to save reproductive effort from early selection caused the evolution of higher deactivation rates and lower activation rates than in the late selection condition. Thus, we predict that in the special case where early selection can save maternal investment in non-viable offspring, gene expression activation rates and deactivation rates might be selected to permit sampling of the product from each allele.


A direct interaction between G-protein beta gamma subunits and the Raf-1 protein kinase.
Raf-1 is a serine/threonine protein kinase positioned downstream of Ras in the mitogen-activated protein kinase cascade. Using a yeast two-hybrid strategy to identify other proteins that interact with and potentially regulate Raf-1, we isolated a clone encoding the carboxyl-terminal half of the G beta 2 subunit of heterotrimeric G-proteins. In vitro, purified G beta gamma subunits specifically bound to a GST fusion protein encoding amino acids 1-330 of Raf-1 (Raf/330). Binding assays with truncation mutants of GST-Raf indicate that the region located between amino acids 136 and 239 is a primary determinant for interaction with G beta gamma. In competition experiments, the carboxyl terminus of beta-adrenergic receptor kinase (beta ARK) blocked the binding of G beta gamma to Raf/330; however, the Raf-1-binding proteins, Ras and 14-3-3, had no effect. Scatchard analysis of in vitro binding between Raf/330 and G beta gamma revealed an affinity of interaction (Kd = 163 +/- 36 nM), similar to that seen between G beta gamma and beta ARK (Kd = 87 +/- 24 nM). The formation of native heterotrimeric G alpha beta gamma complexes, as measured by pertussis toxin ADP-ribosylation of G alpha, could be disrupted by increasing amounts of Raf/330, with an EC50 of approximately 200 nM, in close agreement with the estimated binding affinity. In vivo complexes of Raf-1 and G beta gamma were isolated from human embryonic kidney 293-T cells transfected with epitope-tagged G beta 2. The identification and characterization of this novel interaction raises several possibilities for signaling cross-talk between growth factor receptors and those receptors coupled to heterotrimeric G-proteins.


Fluorescence labelling of extracellular vesicles using a novel thiol-based strategy for quantitative analysis of cellular delivery and intracellular traffic.
Extracellular vesicles, including exosomes, are naturally derived nanovesicles generated in and released by numerous cell types. As extracellular entities they have the capacity to interact with neighbouring cells and distant tissues and affect physiological processes as well as being implicated in numerous diseases including tumorigenesis and neurodegeneration. They are also under intense investigation as delivery vectors for biotherapeutics. The ways in which EVs interact with recipient cells to influence cell physiology and deliver a macromolecular payload are at the early stages of exploration. A significant challenge within these studies is the ability to label EVs directly or indirectly with fluorescent probes to allow visualization without compromising functionality. Here, we present a thiol-based fluorescence labelling method allowing comprehensive analysis of the cellular uptake of prostate cancer derived EVs in live cells using confocal microscopy. Labelling of the EVs in this way did not influence their size and had no effect on their ability to induce differentiation of lung fibroblasts to myofibroblasts. For endocytosis analyses, depletion of key endocytic proteins and the use of chemical inhibitors (Dynasore, EIPA, Rottlerin and IPA-3) indicated that fluid-phase endocytosis and/or macropinocytosis was involved in EV internalisation. Over a period of six hours EVs were observed to increasingly co-localise with lysosomes, indicating a possible termination point following internalisation. Overall this method provides new opportunities for analysing the cellular dynamics of EVs as biological entities affecting cell and whole body physiology as well as investigating their potential as drug delivery vectors.


JACKDAW controls epidermal patterning in the Arabidopsis root meristem through a non-cell-autonomous mechanism.
In Arabidopsis, specification of the hair and non-hair epidermal cell types is position dependent, in that hair cells arise over clefts in the underlying cortical cell layer. Epidermal patterning is determined by a network of transcriptional regulators that respond to an as yet unknown cue from underlying tissues. Previously, we showed that JACKDAW (JKD), a zinc finger protein, localizes in the quiescent centre and the ground tissue, and regulates tissue boundaries and asymmetric cell division by delimiting SHORT-ROOT movement. Here, we provide evidence that JKD controls position-dependent signals that regulate epidermal-cell-type patterning. JKD is required for appropriately patterned expression of the epidermal cell fate regulators GLABRA2, CAPRICE and WEREWOLF. Genetic interaction studies indicate that JKD operates upstream of the epidermal patterning network in a SCRAMBLED (SCM)-dependent fashion after embryogenesis, but acts independent of SCM in embryogenesis. Tissue-specific induction experiments indicate non-cell-autonomous action of JKD from the underlying cortex cell layer to specify epidermal cell fate. Our findings are consistent with a model where JKD induces a signal in every cortex cell that is more abundant in the hair cell position owing to the larger surface contact of cells located over a cleft.


Injury surveillance in multi-sport events: the International Olympic Committee approach.
The protection of athletes' health by preventing injuries is an important task for international sports federations. Standardised injury surveillance provides not only important epidemiological information, but also directions for injury prevention, and the opportunity for monitoring long-term changes in the frequency and circumstances of injury. Numerous studies have evaluated sports injuries during the season, but few have focused on injuries during major sport events such as World Championships, World Cups or the Olympic Games. To provide an injury surveillance system for multi-sports tournaments, using the 2008 Olympic Games in Beijing as an example. A group of experienced researchers reviewed existing injury report systems and developed a scientific sound and concise injury surveillance system for large multi-sport events. The injury report system for multi-sport events is based on an established system for team sports tournaments and has proved feasible for individual sports during the International Association of Athletics Federations World Championships in Athletics 2007. The most important principles and advantages of the system are comprehensive definition of injury, injury report by the physician responsible for the athlete, a single-page report of all injuries, and daily report irrespective of whether or not an injury occurred. Implementation of the injury surveillance system, all definitions, the report form, and the analysis of data are described in detail to enable other researchers to implement the injury surveillance system in any sports tournament. The injury surveillance system has been accepted by experienced team physicians and shown to be feasible for single-sport and multi-sport events. It can be modified depending on the specific objectives of a certain sport or research question; however, a standardised use of injury definition, report forms and methodology will ensure the comparability of results.


Evaluation of plant and fungal extracts for their potential antigingivitis and anticaries activity.
The link between diet and health has lead to the promotion of functional foods which can enhance health. In this study, the oral health benefits of a number of food homogenates and high molecular mass and low molecular mass fractions were investigated. A comprehensive range of assays were performed to assess the action of these foods on the development of gingivitis and caries using bacterial species associated with these diseases. Both antigingivitis and anticaries effects were investigated by assays examining the prevention of biofilm formation and coaggregation, disruption of preexisting biofilms, and the foods' antibacterial effects. Assays investigating interactions with gingival epithelial cells and cytokine production were carried out to assess the foods' anti- gingivitis properties. Anti-caries properties such as interactions with hydroxyapatite, disruption of signal transduction, and the inhibition of acid production were investigated. The mushroom and chicory homogenates and low molecular mass fractions show promise as anti-caries and anti-gingivitis agents, and further testing and clinical trials will need to be performed to evaluate their true effectiveness in humans.


Role of Substrate in Au Nanoparticle Decoration by Electroless Deposition.
Decoration of nanostructures is a promising way of improving performances of nanomaterials. In particular, decoration with Au nanoparticles is considerably efficient in sensing and catalysis applications. Here, the mechanism of decoration with Au nanoparticles by means of low-cost electroless deposition (ELD) is investigated on different substrates, demonstrating largely different outcomes. ELD solution with Au potassium cyanide and sodium hypophosphite, at constant temperature (80 °C) and pH (7.5), is used to decorate by immersion metal (Ni) or semiconductor (Si, NiO) substrates, as well as NiO nanowalls. All substrates were pre-treated with a hydrazine hydrate bath. Scanning electron microscopy and Rutherford backscattering spectrometry were used to quantitatively analyze the amount, shape and size of deposited Au. Au nanoparticle decoration by ELD is greatly affected by the substrates, leading to a fast film deposition onto metallic substrate, or to a slow cluster (50-200 nm sized) formation on semiconducting substrate. Size and density of resulting Au clusters strongly depend on substrate material and morphology. Au ELD is shown to proceed through a galvanic displacement on Ni substrate, and it can be modeled with a local cell mechanism widely affected by the substrate conductivity at surface. These data are presented and discussed, allowing for cheap and reproducible Au nanoparticle decoration on several substrates.


Optimal control of molecular fragmentation with homologous families of photonic reagents and chemical substrates.
This work investigates laser optimally controlled fragmentation in a series of chemically related halomethane compounds in order to determine the extent to which regular trends in objective yields are observed upon variation of both the laser pulse shape and the chemical substrate. Three control objectives defined by ratios of fragment ions were considered, where families of shaped pulses were found that produced optimal objective yields. For the objective defined by the ratio of a halogen ion to a methyl halide ion, a systematic correlation between the optimal objective yield and chemical composition was revealed. Furthermore, some of the optimal shaped pulses were found to successfully control the same objective in closely related molecular substrates. These results provide a basis to expect systematic chemical responses from optimally shaped laser pulses acting as "photonic reagents" in analogy with the action of traditional chemical reagents.


Maternal B vitamin intake during pregnancy and wheeze and eczema in Japanese infants aged 16-24 months: the Osaka Maternal and Child Health Study.
It is uncertain whether B group vitamins are risk or preventive factors for allergic disorders. We prospectively investigated the association between maternal intake of folate and vitamins B(12) , B(6) , and B(2) during pregnancy and the risk of wheeze and eczema in the infants aged 16-24 months. Subjects were 763 Japanese mother-child pairs. Data on maternal intake during pregnancy were assessed with a diet history questionnaire (DHQ). Symptoms of wheeze and eczema were based on criteria of the International Study of Asthma and Allergies in Childhood. Among 763 infants, 169 (22.1%) and 142 (18.6%) had symptoms of wheeze and eczema, respectively. There were no evident relationships between maternal consumption of folate, vitamin B(12) , vitamin B(6) , and vitamin B(2) during pregnancy and the risk of wheeze or eczema in the offspring after adjustment for maternal age, gestation at baseline, residential municipality at baseline, family income, maternal and paternal education, maternal and paternal history of asthma, atopic eczema, and allergic rhinitis, changes in maternal diet in the previous 1 month, season when data at baseline were collected, maternal smoking during pregnancy, baby's older siblings, baby's sex, baby's birth weight, household smoking in the same room as the infant, breastfeeding duration, age at which solid foods were introduced, age of infant at the third survey, and maternal intake of docosahexaenoic acid, n-6 polyunsaturated fatty acids, vitamin D, calcium, vitamin E, and β-carotene during pregnancy. Further investigation is warranted to draw conclusions as to the question of whether maternal B vitamin intake during pregnancy is related to the risk of childhood allergic disorders.


A Survey on Learning to Hash.
Nearest neighbor search is a problem of finding the data points from the database such that the distances from them to the query point are the smallest. Learning to hash is one of the major solutions to this problem and has been widely studied recently. In this paper, we present a comprehensive survey of the learning to hash algorithms, categorize them according to the manners of preserving the similarities into: pairwise similarity preserving, multiwise similarity preserving, implicit similarity preserving, as well as quantization, and discuss their relations. We separate quantization from pairwise similarity preserving as the objective function is very different though quantization, as we show, can be derived from preserving the pairwise similarities. In addition, we present the evaluation protocols, and the general performance analysis, and point out that the quantization algorithms perform superiorly in terms of search accuracy, search time cost, and space cost. Finally, we introduce a few emerging topics.


Global analysis of Plasmodium falciparum histidine-rich protein-2 (pfhrp2) and pfhrp3 gene deletions using whole-genome sequencing data and meta-analysis.
Many rapid diagnostic tests (RDT) used on suspected malaria cases are based on the detection of the protein encoded by the Plasmodium falciparum histidine-rich protein-2 (pfhrp2) gene, which shares a high sequence homology with pfhrp3 in the 3D7 reference genome. Parasite isolates showing pfhrp2 and pfhrp3 gene deletions have been emerging over the years, but a comprehensive genetic analysis of these variants is still lacking. With this purpose, genomic data from experimental P. falciparum genetic crosses between different laboratory lines (3D7, HB3, DD2, 7G8 and GB4) were first analysed (n = 98). The frequency of pfhrp2 deletions was consistent with a Mendelian prediction in HB3 × DD2 (56.7%; 95%CI = (39.5%-72.9%)). Moreover, the pfhrp2 and pfhrp3 deletions segregated independently of each other in the same genetic cross. Analysis of 3D7 × HB3 and 7G8 × GB4 estimated the probability of spontaneously generating a pfhrp2 deletion during sexual recombination to be up to 6.2%. Next, whole genome sequence data from 1970 P. falciparum isolates collected globally were analysed. Nine samples displayed depth of coverage consistent with pfhrp2 deletions (0.5%), but the corresponding split-read analysis could not confirm deletions in seven of these samples. Twenty-eight isolates had evidence of pfhrp3 deletions (1.4%), which are widespread in Southeast Asia. Finally, a meta-analysis of published data revealed a positive mean association between the frequencies of pfhrp2 and pfhrp3 deletions in Africa and South America. This result suggested a shared selective pressure acting on these genetic variants. In conclusion, evidence of genetic selection on both pfhrp2 and pfhrp3 deletions was presented, but experimental crosses do not provide evidence of a fitness cost of these variants. Further work is urgently needed to accurately determine the prevalence and the degree of association between these genetic variants, and the respective impact on diagnostic accuracy of many in-use RDT.


Impact of solvent quality on the network strength and structure of alginate gels.
The influence of the mixture of water and alcohols on the solubility and properties of alginate and its calcium-induced gels is of interest for the food, wound care and pharmaceutical industries. The solvent quality of water with increasing amounts of ethanol (0-20%) on alginate was studied using intrinsic viscosity. The effect of ethanol addition on the rheological and mechanical properties of calcium alginate gels was determined. Small-angle X-ray scattering and transmission electron microscopy were used to study the network structure. It is shown that the addition of ethanol up to 15% (wt) increases the extension of the alginate chain, which correlates with increased moduli and stress being required to fracture the gels. The extension of the polymer chain is reduced at 20% (wt) ethanol, which is followed by reduced moduli and stress at breakage of the gels. The network structure of gels at high ethanol concentrations (24%) is characterized by thick and poorly connected network strands.


A multiscale expectation-maximization semisupervised classifier suitable for badly posed image classification.
This paper deals with the problem of badly posed image classification. Although underestimated in practice, bad-posedness is likely to affect many real-world image classification tasks, where reference samples are difficult to collect (e.g., in remote sensing (RS) image mapping) and/or spatial autocorrelation is relevant. In an image classification context affected by a lack of reference samples, an original inductive learning multiscale image classifier, termed multiscale semisupervised expectation maximization (MSEM), is proposed. The rationale behind MSEM is to combine useful complementary properties of two alternative data mapping procedures recently published outside of image processing literature, namely, the multiscale modified Pappas adaptive clustering (MPAC) algorithm and the sample-based semisupervised expectation maximization (SEM) classifier. To demonstrate its potential utility, MSEM is compared against nonstandard classifiers, such as MPAC, SEM and the single-scale contextual SEM (CSEM) classifier, besides against well-known standard classifiers in two RS image classification problems featuring few reference samples and modestly useful texture information. These experiments yield weak (subjective) but numerous quantitative map quality indexes that are consistent with both theoretical considerations and qualitative evaluations by expert photointerpreters. According to these quantitative results, MSEM is competitive in terms of overall image mapping performance at the cost of a computational overhead three to six times superior to that of its most interesting rival, SEM. More in general, our experiments confirm that, even if they rely on heavy class-conditional normal distribution assumptions that may not be true in many real-world problems (e.g., in highly textured images), semisupervised classifiers based on the iterative expectation maximization Gaussian mixture model solution can be very powerful in practice when: 1) there is a lack of reference samples with respect to the problem/model complexity and 2) texture information is considered negligible (i.e., a piecewise constant image model holds).


Atypical depressive symptoms and obesity in a national sample of older adults with major depressive disorder.
The objectives of this study are to present findings on the rate of obesity associated with classic, atypical, and undifferentiated depression by comparing with those without depression in a nationally representative sample of United States older adults. The authors used data from the 2001 to 2002 National Epidemiologic Survey of Alcohol and Related Conditions (NESARC), which included 10,557 adults 60 years of age and older. Chi-square tests were used to compare classic, atypical, and undifferentiated as well as nondepressed control in sociodemographic characteristics. Then, logistic regressions adjusting for sociodemographic characteristics were used to evaluate associations of rate of current obesity (defined as Body Mass Index (BMI) > 30) across the three depressive groups (classic, atypical, and undifferentiated depression) and nondepressed control. Lifetime, current, and past depression were examined. Significant differences were found between atypical and classic depression in sex, age, marital status, race, and personal income. After adjusting for sex, age, marital status, race, and personal income, the rate of obesity was significantly greater for respondents with atypical depression than respondents with classic, undifferentiated depression, or without depression. Same results were found in lifetime, current, and past depression. Our findings suggest that the heterogeneity of depression should be considered when examining the effect of depression on obesity in old age. Prevention measures should be designed and delivered to older adults with atypical depression.


Organizational metrics of interchromatin speckle factor domains: integrative classifier for stem cell adhesion & lineage signaling.
Stem cell fates on biomaterials are influenced by the complex confluence of microenvironmental cues emanating from soluble growth factors, cell-to-cell contacts, and biomaterial properties. Cell-microenvironment interactions influence the cell fate by initiating a series of outside-in signaling events that traverse from the focal adhesions to the nucleus via the cytoskeleton and modulate the sub-nuclear protein organization and gene expression. Here, we report a novel imaging-based framework that highlights the spatial organization of sub-nuclear proteins, specifically the splicing factor SC-35 in the nucleoplasm, as an integrative marker to distinguish between minute differences of stem cell lineage pathways in response to stimulatory soluble factors, surface topologies, and microscale topographies. This framework involves the high resolution image acquisition of SC-35 domains and imaging-based feature extraction to obtain quantitative nuclear metrics in tandem with machine learning approaches to generate a predictive cell state classification model. The acquired SC-35 metrics led to >90% correct classification of emergent human mesenchymal stem cell (hMSC) phenotypes in populations of hMSCs exposed for merely 3 days to basal, adipogenic, or osteogenic soluble cues, as well as varying levels of dexamethasone-induced alkaline phosphatase (ALP) expression. Early osteogenic cellular responses across a series of surface patterns, fibrous scaffolds, and micropillars were also detected and classified using this imaging-based methodology. Complex cell states resulting from inhibition of RhoGTPase, β-catenin, and FAK could be classified with >90% sensitivity on the basis of differences in the SC-35 organizational metrics. This indicates that SC-35 organization is sensitively impacted by adhesion-related signaling molecules that regulate osteogenic differentiation. Our results show that diverse microenvironment cues affect different attributes of the SC-35 organizational metrics and lead to distinct emergent organizational patterns. Taken together, these studies demonstrate that the early organization of SC-35 domains could serve as a "fingerprint" of the intracellular mechanotransductive signaling that governs growth factor- and topography-responsive stem cell states.


Hydrodynamics of shape-driven rigidity transitions in motile tissues.
In biological tissues, it is now well-understood that mechanical cues are a powerful mechanism for pattern regulation. While much work has focused on interactions between cells and external substrates, recent experiments suggest that cell polarization and motility might be governed by the internal shear stiffness of nearby tissue, deemed "plithotaxis". Meanwhile, other work has demonstrated that there is a direct relationship between cell shapes and tissue shear modulus in confluent tissues. Joining these two ideas, we develop a hydrodynamic model that couples cell shape, and therefore tissue stiffness, to cell motility and polarization. Using linear stability analysis and numerical simulations, we find that tissue behavior can be tuned between largely homogeneous states and patterned states such as asters, controlled by a composite "morphotaxis" parameter that encapsulates the nature of the coupling between shape and polarization. The control parameter is in principle experimentally accessible, and depends both on whether a cell tends to move in the direction of lower or higher shear modulus, and whether sinks or sources of polarization tend to fluidize the system.


Brain morphology of childhood aggressive behavior: A multi-informant study in school-age children.
Few studies have focused on the neuroanatomy of aggressive behavior in children younger than 10 years. Here, we explored the neuroanatomical correlates of aggression in a population-based sample of 6- to 9-year-old children using a multiple-informant approach. Magnetic resonance (MR) scans were acquired from 566 children from the Generation R study who participated in the Berkeley Puppet Interview and whose parents had completed the Child Behavior Checklist. Linear regression analyses were used to examine associations between aggression and amygdala and hippocampal volume. We performed surface-based analyses to study the association between aggression and cortical thickness, surface area, and gyrification. Aggressive behavior was associated with smaller amygdala (p < .05) but not hippocampal volume. Aggression was associated with a thinner cortex in the left precentral cortex (p < .01) and in a cluster including the right inferior parietal, supramarginal, and postcentral cortex (p < .001). Gender moderated the association between aggression and cortical thickness in the right medial posterior cortex (p = .001) and the right prefrontal cortex (p < .001). Aggression was associated with decreased gyrification in a large cluster including the right precentral, postcentral, frontal, and parietal cortex (p = .01). Moreover, aggression was associated with decreased gyrification in the right occipital and parietal cortex (p = .02). We found novel evidence that childhood aggressive behavior is related to decreased amygdala volume, decreased sensorimotor cortical thickness, and decreased global right hemisphere gyrification. Aggression is related to cortical thickness in regions associated with the default mode network, with negative associations in boys and positive associations in girls.


Demon rum: high-tech solutions to an age-old problem.
This article summarizes the proceedings of a symposium at the 2004 Research Society on Alcoholism Meeting in Vancouver, British Columbia, Canada, organized and chaired by Scott T. Walters. The purpose of the symposium was to describe several brief motivational interventions offered via the Internet, including the evidence for web-based interventions, applications and contexts in which such approaches are being used, and directions for future research. Walters provided an overview and introduction to the topic and discussed the e-CHUG (www.e-chug.com) and e-TOKE (www.e-toke.com) feedback interventions for college alcohol and marijuana prevention, including the contexts in which they are being used and ways they are being integrated with other campus prevention efforts. Dr. Hester presented 12-month results from a controlled trial of the Drinker's Check-up (www.drinkerscheckup.com), an intervention for adult problem drinkers that is available both as a Windows and as an Internet application. Dr. Chiauzzi described the development and testing of My Student Body (www.mystudentbody.com), a tailored drinking prevention web site for college students. Finally, Dr. Miller addressed the use of online assessment and feedback to reduce drinking, including the history of web-based interventions and their likely future and the potential limitations of such approaches.


Pain-related anxiety and anxiety sensitivity across anxiety and depressive disorders.
Fear-anxiety-avoidance models posit pain-related anxiety and anxiety sensitivity as important contributing variables in the development and maintenance of chronic musculoskeletal pain [Asmundson, G. J. G, Vlaeyen, J. W. S., & Crombez, G. (Eds.). (2004). Understanding and treating fear of pain. New York: Oxford University Press]. Emerging evidence also suggests that pain-related anxiety may be a diathesis for many other emotional disorders [Asmundson, G. J. G., & Carleton, R. N. (2005). Fear of pain is elevated in adults with co-occurring trauma-related stress and social anxiety symptoms. Cognitive Behaviour Therapy, 34, 248-255; Asmundson, G. J. G., & Carleton, R. N. (2008). Fear of pain. In: M. M. Antony & M. B. Stein (Eds.), Handbook of anxiety and the anxiety disorders (pp. 551-561). New York: Oxford University Press] and appears to share several elements in common with other fears (e.g., anxiety sensitivity, illness/injury sensitivity, fear of negative evaluation) as described by Reiss [Reiss, S. (1991). Expectancy model of fear, anxiety, and panic. Clinical Psychology Review, 11, 141-153] and Taylor [Taylor, S. (1993). The structure of fundamental fears. Journal of Behavior Therapy and Experimental Psychiatry, 24, 289-299]. The purpose of the present investigation was to assess self-reported levels of pain-related anxiety [Pain Anxiety Symptoms Scale-Short Form; PASS-20; McCracken, L. M., & Dhingra, L. (2002). A short version of the Pain Anxiety Symptoms Scale (PASS-20): preliminary development and validity. Pain Research and Management, 7, 45-50] across several anxiety and depressive disorders and to compare those levels to non-clinical and chronic pain samples. Participants consisted of a clinical sample (n=418; 63% women) with principal diagnoses of a depressive disorder (DD; n=22), panic disorder (PD; n=114), social anxiety disorder (SAD; n=136), obsessive-compulsive disorder (OCD; n=86), generalized anxiety disorder (GAD; n=46), or specific phobia (n=14). Secondary group comparisons were made with a community sample as well as with published data from a treatment-seeking chronic pain sample [McCracken, L. M., & Dhingra, L. (2002). A short version of the Pain Anxiety Symptoms Scale (PASS-20): preliminary development and validity. Pain Research and Management, 7, 45-50]. Results suggest that pain-related anxiety is generally comparable across anxiety and depressive disorders; however, pain-related anxiety was typically higher (p<.01) in individuals with anxiety and depressive disorders relative to a community sample, but comparable to or lower than a chronic pain sample. Results imply that pain-related anxiety may indeed be a construct independent of other fundamental fears, warranting subsequent hierarchical investigations and consideration for inclusion in treatments of anxiety disorders. Additional implications and directions for future research are discussed.


Regulation of the catalytic activity of the human phosphatase PTPN4 by its PDZ domain.
The human protein tyrosine phosphatase non-receptor type 4 (PTPN4) prevents cells death. Targeting its PDZ domain abrogates this protection and triggers apoptosis. We demonstrate here that the PDZ domain inhibits the phosphatase activity of PTPN4. The mere binding of a PDZ ligand is sufficient to release the catalytic inhibition. We combined analytical ultracentrifugation, small angle X-ray scattering and NMR to understand how the PDZ domain controls PTPN4 activity. We show that the physiologically active PTPN4 two-domain, encompassing the PDZ and the phosphatase domains, adopts a predominant compact conformation in solution. The PDZ ligand binding restores the catalytic competence of PTPN4 disrupting the transient interdomain communication. This study strengthens the emerging notion that PDZ domains can act as regulators of enzyme activity and therefore are active players in the dynamic regulation of signaling pathways.


Twins in Jos Nigeria: a seven-year retrospective study.
A retrospective analysis of twin pregnancy was carried out between 1st January 1992 and 31st December 1998 to determine the incidence, epidemiological variables perinatal and maternal outcome of twin pregnancy in the institution. The case notes of the 413 twin pregnancy, 6 triplets and 1 quadruplet delivered during the period, and the labour records of all the deliveries were analyzed. Statistical analysis was with chi-squared test. The level of significance was set at 5%. The incidence of twin pregnancy was 28/1000, triplet 0.4/1000 and quadruplet 0.07/1000. The age range of the twin mothers was 15-47 years with a mean of 28 +/- 5.4 years. The parity range was 0-14 years with a mean of 3.3 +/- 2.6. Rising maternal age and parity were associated with twinning rate. The mean fetal weight was 2.21 +/- 0.66 kg. There was no statistical significant difference in weight between the 1st and 2nd twins. Male babies however weighed heavier (p < 0.02) than the female infants. The incidence of low birth weight babies (< 2500 g) was 58.7% with a perinatal mortality rate of 278.4/1000. The perinatal outcome was worst for the very low birth weight babies (< 1500), 817.5/1000 8.5% of twin mothers had a retained 2nd twin. The perinatal outcome of the 27 unbooked mothers who delivered their 1st twin at home was (12) 44.4/1000, compared to the rate of 12.5/1000 (1) in 8 booked mothers, who delivered the 1st twin in the hospital. Among the 771 babies who were alive at the time of presentation in the labour ward, delivery by caesarean section had a better perinatal outcome (6.8%), compared to spontaneous vaginal delivery (13.1%) and assisted breech delivery (21.4%). The unbooked twin mothers had a perinatal mortality rate of 318.7/1000 compared to booked mothers 82.3/1000. The overall perinatal mortality rate was 186.4/1000. The clinical causes of death were prematurity 74.6%, retained 2nd twin 7.3%, antepartum haemorrhage 6.1%, severe pregnancy induced hypertension/eclampsia 3.0%, birth asphyxia 2.4%, congenital malformation 1.8%. The incidence of postpartum haemorrhage was 3% and there was no maternal mortality in the series. The institution will need to put in place adequate resuscitating facilities and manpower to manage the low birth weight infants so as to reduce the current high perinatal wastage.


Antinociceptive effect of selective G protein-gated inwardly rectifying K+ channel agonist ML297 in the rat spinal cord.
The G protein-gated inwardly rectifying K+ (GIRK) channels play important signaling roles in the central and peripheral nervous systems. However, the role of GIRK channel activation in pain signaling remains unknown mainly due to the lack of potent and selective GIRK channel activators until recently. The present study was designed to determine the effects and mechanisms of ML297, a selective GIRK1/2 activator, on nociception in the spinal cord by using behavioral studies and whole-cell patch-clamp recordings from substantia gelatinosa (SG) neurons. Rats were prepared for chronic lumber catheterization and intrathecal administration of ML297. The nociceptive flexion reflex was tested using an analgesy-meter, and the influence on motor performance was assessed using an accelerating rotarod. We also investigated pre- and post-synaptic actions of ML297 in spinal cord preparations by whole-cell patch-clamp recordings. Intrathecal administration of ML297 increased the mechanical nociceptive threshold without impairing motor function. In voltage-clamp mode of patch-clamp recordings, bath application of ML297 induced outward currents in a dose-dependent manner. The ML297-induced currents demonstrated specific equilibrium potential like other families of potassium channels. At high concentration, ML297 depressed miniature excitatory postsynaptic currents (mEPSCs) but not their amplitude. The ML297-induced outward currents and suppression of mEPSCs were not inhibited by naloxone, a μ-opioid receptor antagonist. These results demonstrated that intrathecal ML297 showed the antinociceptive effect, which was mediated through direct activation of pre- and post-synaptic GIRK channels. Selective GIRK channel activation is a promising strategy for the development of new agents against chronic pain and opioid tolerance.