Most antidepressant use in primary care is justified; results of the Netherlands Study of Depression and Anxiety.
Depression is a common illness, often treated in primary care. Many studies have reported undertreatment with antidepressants in primary care. Recently, some studies also reported overtreatment with antidepressants. The present study was designed to assess whether treatment with antidepressants in primary care is in accordance with current guidelines, with a special focus on overtreatment. We used baseline data of primary care respondents from the Netherlands Study of Depression and Anxiety (NESDA) (n = 1610). Seventy-nine patients with treatment in secondary care were excluded. We assessed justification for treatment with antidepressant according to the Dutch primary care guidelines for depression and for anxiety disorders. Use of antidepressants was based on drug-container inspection or, if unavailable, on self-report. Results were recalculated to the original population of primary care patients from which the participants in NESDA were selected (n = 10,677). Of 1531 included primary care patients, 199 (13%) used an antidepressant, of whom 188 (94.5%) (possibly) justified. After recalculating these numbers to the original population (n = 10,677), we found 908 (95% CI 823 to 994) antidepressant users. Forty-nine (95% CI 20 to 78) of them (5.4%) had no current justification for an antidepressant, but 27 of them (54.5%) had a justified reason for an antidepressant at some earlier point in their life. We found that overtreatment with antidepressants in primary care is not a frequent problem. Too long continuation of treatment seems to explain the largest proportion of overtreatment as opposed to inappropriate initiation of treatment.


Characteristics of community-based occupational therapy: Results of a norwegian survey.
Background: Ongoing changes in healthcare delivery systems in Norway increasingly require community-based services, and the changes will likely affect the working conditions and opportunities for occupational therapists.Aim: To characterize occupational therapy in community-based practice in Norway.Material and methods: A cross-sectional, descriptive survey design was applied using a questionnaire related to personal and organizational characteristics. Participants (n = 561) were recruited among community-working occupational therapists in Norway registered as members of Ergoterapeutene. Data were analyzed with descriptive statistics.Results: The majority of the participants was female and had an average of 16.5 years of professional experience. They reported to spend about half of their working hours on direct work with clients. For many, work with assistive technology was a main task, accounting for approximately half their working hours. Only a small proportion worked in municipalities that had merged with others, but for a larger proportion (27%) a merger had been decided and was in preparation.Conclusion: This study established some basic information regarding Norwegian community-based occupational therapy and the municipalities where occupational therapists work.Significance: With this study serving as a baseline, we may be able to track how changes will affect community-based occupational therapy practice in the near future.


Regulated secretion of multi-lamellar vesicles leads to formation of a tubulo-vesicular network in host-cell vacuoles occupied by Toxoplasma gondii.
Toxoplasma gondii is an obligate intracellular parasite that actively invades virtually all types of nucleated cells, surviving within a specialized vacuole called the parasitophorous vacuole. Shortly after invasion, the parasite modifies this vacuole by secreting a variety of proteins from electron-dense storage granules. Additionally, the parasite forms a network of membranous tubules within the lumen of the vacuole and connecting with the vacuolar membrane. We have used immunolabeling and cell fractionation to examine the secretion of two dense granule proteins, GRA1 and GRA2, which are involved in formation of the intravacuolar network. Following host-cell invasion, GRA1 was secreted into the lumen of the vacuole as a soluble protein that subsequently became peripherally associated with the network. In addition to being secreted as a soluble protein from dense granules, GRA2 was secreted within multi-lamellar vesicles released from a specialized posterior invagination of the parasite. The multi-lamellar vesicles assemble to form the intravacuolar network, which contains an integral membrane form of GRA2. These findings indicate that Toxoplasma has a highly developed regulated exocytosis pathway that modifies the parasitophorous vacuole by secretion of soluble proteins and by a novel process of membrane secretion.


Are medical eponyms really dying out? A study of their usage in the historical biomedical literature.
Eponyms are a prominent feature of medical language. Many feel they have had their time and serve only to complicate medical education and conversation. Others argue that eponyms can make unmemorable concepts memorable, can concisely label complex concepts, and promote a valuable interest in medical history. It is frequently assumed that medical eponyms are marching towards extinction. However, this hypothesis has not been adequately tested. The fate of 8,636 eponyms from contemporary and historical registers is presented here. The annual usage of each eponym in PubMed indexed articles since 1900 is analysed with metrics of eponym prevalence, coinage and extinction. The fate of eponyms named for those implicated in Nazi war crimes is also measured. The analysis shows that more publications used an eponym in their title in 2014 than ever before. However, the proportion using an eponym title has fallen from a peak of 3% in 1991 to 2% today, while annual extinction of eponyms overtook coinage in the 1980s. This suggests that, while not in dramatic decline, they are perhaps losing market share to more scientific descriptions. There is no ambiguity in the fate of eponyms related to Nazi war crimes which have almost entirely fallen out of use in the last decade.


Fine-tuning of NADH oxidase decreases byproduct accumulation in respiration deficient xylose metabolic Saccharomyces cerevisiae.
Efficiently utilizing all available carbon from lignocellulosic feedstock presents a major barrier to the production of economically feasible biofuel. Previously, to enable xylose utilization, we introduced a cofactor-dependent xylose reductase (XR) and xylitol dehydrogenase (XDH) pathway, or a cofactor-independent xylose isomerase (XI) pathway, into Saccharomyces cerevisiae. The resulting strains metabolized xylose with high efficiency. However, in both pathway recombinant strains, the cofactor imbalance caused accumulation of the byproducts glycerol and/or xylitol and reduced the ethanol production efficiency. In this study, we introduced NADH oxidase from Lactococcus lactis into both XI and XR-XDH pathway recombinant strains. To reduce byproduct accumulation while maintaining xylose metabolism, we optimized the expression level of NADH oxidase by comparing its expression under the control of different promoters and plasmids. In recombinant XI strains, NADH oxidase was expressed at different levels, regulated by the GPD2 promoter or TEF1 promoter in the 2 μ plasmid. The expression under the control of GPD2 promoter decreased glycerol production by 84% and increased the ethanol yield and specific growth rate by 8% and 12%, respectively. In contrast, in the recombinant XR-XDH strains, such expression level was not efficient enough to decrease the byproduct accumulation. Therefore, higher NADH oxidase expression levels were tested. In the strain expressing NADH oxidase under the control of the TEF1 promoter in the centromeric plasmids, xylitol and glycerol production were reduced by 60% and 83%, respectively, without significantly affecting xylose consumption. By fine-tuning NADH oxidase expression, we decreased the glycerol or/and xylitol production in both recombinant XI and XR-XDH xylose-metabolizing yeast strains. The optimal NADH oxidase expression levels depend on metabolic pathways. Similar cofactor engineering strategies could maximize the production of other redox dependent metabolites.


Mitosis can drive cell cannibalism through entosis.
Entosis is a form of epithelial cell cannibalism that is prevalent in human cancer, typically triggered by loss of matrix adhesion. Here, we report an alternative mechanism for entosis in human epithelial cells, driven by mitosis. Mitotic entosis is regulated by Cdc42, which controls mitotic morphology. Cdc42 depletion enhances mitotic deadhesion and rounding, and these biophysical changes, which depend on RhoA activation and are phenocopied by Rap1 inhibition, permit subsequent entosis. Mitotic entosis occurs constitutively in some human cancer cell lines and mitotic index correlates with cell cannibalism in primary human breast tumours. Adherent, wild-type cells can act efficiently as entotic hosts, suggesting that normal epithelia may engulf and kill aberrantly dividing neighbours. Finally, we report that Paclitaxel/taxol promotes mitotic rounding and subsequent entosis, revealing an unconventional activity of this drug. Together, our data uncover an intriguing link between cell division and cannibalism, of significance to both cancer and chemotherapy.


Cerebral Venous Thrombosis in the Mediterranean Area in Adults. Role of Behçet's Disease as an Underlying Cause.
Cerebral venous and dural sinus thrombosis (CVT) is a rare condition with a wide spectrum of clinical presentations. The epidemiology of the disease has evolved considerably during the recent decades with increasing oral contraceptive use in young and middle-aged women. CVT has various causes including genetic and acquired prothrombotic disorders and it usually has a favorable outcome with a low rate of thrombotic recurrence and mortality. Geographical and ethnic variations between populations may result in different distribution of CVT etiologies leading to different pathophysiological mechanisms and clinical presentations. In CVT series reported mostly from the Americas and the western European countries Behçet's disease (BD) is not reported as a common cause of CVT. However it can be discerned as a frequent cause of CVT in BD series. Due to the high prevalence of BD in the southeast Mediterranean region BD is a frequent cause of CVT in the area. Discerning characteristics of patients with BD and CVT have been reported previously and these might be helpful in guiding diagnosis and treatment of CVT especially in this part of the world.


PIM1 accelerates prostate cancer cell motility by phosphorylating actin capping proteins.
The PIM family kinases promote cancer cell survival and motility as well as metastatic growth in various types of cancer. We have previously identified several PIM substrates, which support cancer cell migration and invasiveness. However, none of them are known to regulate cellular movements by directly interacting with the actin cytoskeleton. Here we have studied the phosphorylation-dependent effects of PIM1 on actin capping proteins, which bind as heterodimers to the fast-growing actin filament ends and stabilize them. Based on a phosphoproteomics screen for novel PIM substrates, we have used kinase assays and fluorescence-based imaging techniques to validate actin capping proteins as PIM1 substrates and interaction partners. We have analysed the functional consequences of capping protein phosphorylation on cell migration and adhesion by using wound healing and real-time impedance-based assays. We have also investigated phosphorylation-dependent effects on actin polymerization by analysing the protective role of capping protein phosphomutants in actin disassembly assays. We have identified capping proteins CAPZA1 and CAPZB2 as PIM1 substrates, and shown that phosphorylation of either of them leads to increased adhesion and migration of human prostate cancer cells. Phosphorylation also reduces the ability of the capping proteins to protect polymerized actin from disassembly. Our data suggest that PIM kinases are able to induce changes in actin dynamics to support cell adhesion and movement. Thus, we have identified a novel mechanism through which PIM kinases enhance motility and metastatic behaviour of cancer cells. Video abstract.


Reactivity of bulky formamidinatosamarium(II or III) complexes with C═O and C═S bonds.
The preparation of a new heterobimetallic samarium(II) formamidinate complex and selected reactions of samarium(II) complexes and one samarium(III) formamidinate complex with benzophenone or CS2 are discussed. Treatment of the tris(formamidinato)samarium(III) complex [Sm(DippForm)3] 1 (DippForm = N,N'-bis(2,6-diisopropylphenyl)formamidinate, (CH(NC6H3-(i)Pr2-2,6)2) with potassium graphite in toluene yielded the dark brown heterobimetallic formamidinatosamarium(II)/potassium complex [KSm(DippForm)3]n 2. Divalent 2, a Lewis base solvent free homoleptic species, differs significantly from the related heteroleptic formamidinatosamarium(II) complex [Sm(DippForm)2(thf)2] 3 with respect to its constitution, structure, and reactivity toward benzophenone. While 2 reacts giving complex 1, the reaction of 3 with benzophenone generates the highly unusual [Sm(DippForm)2(thf){μ-OC(Ph)═(C6H5)C(Ph)2O}Sm(DippForm)2] (C6H5 = 1,4-cyclohexadiene-3-yl-6-ylidene) 4. The formation of 4 highlights a rare C-C coupling between a carbonyl carbon and the carbon at the para position of a phenyl group of the OCPh2 fragment. An analogous reaction of [Yb(DippForm)2(thf)2] gives an isostructural complex 4Yb. 3 reacts with carbon disulfide forming a light green dinuclear formamidinatosamarium(III) complex [{Sm(DippForm)2(thf)}2(μ-η(2)(C,S):κ(S',S″)-SCSCS2)] 5 through an unusual C-S coupling induced by an amidinatolanthanoid species giving the thioformylcarbonotrithioate ligand. The trivalent organometallic [Sm(DippForm)2(CCPh)(thf)] complex activates the C═O bond of benzophenone by an insertion reaction, forming the light yellow [Sm(DippForm)2{OC(Ph)2C2Ph}(thf)] 6 as a major product and light yellow unsolvated [Sm(DippForm)2{OC(Ph)2C2Ph}] 7 as a minor product. Molecular structures of complexes (2, 4-7) show that κ(N,N') bonding between a DippForm and samarium atom exists in all compounds, but in 2, DippForm also bridges K and Sm by 1κ(N):2κ(N') bonding and two 2,6-diisopropylphenyl groups are η(6)-bonded to potassium.


Changes in frontal morphology after single-stage open posterior-middle vault expansion for sagittal craniosynostosis.
There is controversy regarding whether the frontal bossing associated with sagittal synostosis requires direct surgical correction or spontaneously remodels after isolated posterior cranial expansion. The authors retrospectively measured changes in frontal bone morphology in patients with isolated sagittal synostosis 2 years after open posterior and midvault cranial expansion and compared these changes with those occurring in age-comparable healthy control groups. Forty-three patients age 1 year or younger (mean, 6 months) with sagittal synostosis underwent computed tomography scan digital analysis immediately after and 2 years after posterior-middle cranial vault expansion. Quantitative angular and linear measures were taken along the midsagittal and axial planes to capture both aspects of frontal bossing. The change in values over the 2 years were compared with healthy controls with normal computed tomography scans taken to rule out head trauma. All measures indicative of frontal bossing decreased significantly from the time of posterior-middle vault expansion to 2 years postoperatively. Whereas the majority of patients at time of the operation had frontal bossing measures greater than two standard deviations outside the age-comparable control mean, almost all patients were within two standard deviations of the norm 2 years later. Lateral forehead bossing and anterior cranial growth was greater the older the patient was at the time of the operation, suggesting that the more time that passed before the operation, the more compensatory anterior fossa growth occurred. Central forehead position relative to the anterior cranial base was greatest in the younger patients at the time of operation, suggesting that a central forehead bulge was an early compensatory response to premature sagittal fusion. As a group, patients with sagittal synostosis start to normalize their forehead morphology within 2 years if an isolated posterior operation is performed at 1 year of age or younger, and this occurs by a combination of restriction of growth and reduction relative to patients without synostosis. This protocol decreases the risks of intraoperative positioning, forehead contour deformities, and two-stage operations. Therapeutic, III.


[Treating traumatic ossicular chain disruption with epitympanum approach following tympanotomy].
To evaluate the therapeutic effects of epitympanum approach tympanotomy in treating traumatic ossicular chain disruption. Ten patients with temporal bone fracture and trauma of ear, whose CT scan of temporal bone showed that Malleoincudal joint irregular shape in the axial view and incudostapedial joint dislocation in the coronal view, were included in this study, Virtual endoscopy showed Malleoincudal joint separation, incudostapedial joint dislocation and interruption, ossicular chain disruption in all patients. The surgical approach was opening the atticus via temporal line, remaining the intact posterior and superior wall of external acoustic meatus, exposing the atticus and posterior tympanic cavity sufficiently, and treating the lesions of ossicular chain. One patient with the long process of the incus fracture, adopted the fitted incus prosthesis between the handle of the malleus and the head of the stapes. Six patients with Malleoincudal joint separation or incudostapedial joint dislocation, 3 patients with incus body dislocation completely, adopted incus positioning two articulation decis. The air-conducting hearing levels had an enhancement of 25-50dB in average after surgery. The effect of intact canal wall epitympanum approach ossicular reposition in treating traumatic ossicular chain disruption is satisfying.


Curative Embolization of Arteriovenous Malformations.
Arteriovenous malformations have a significant cumulative risk for hemorrhage. Treatment options include observation, microsurgical resection, stereotactic radiosurgery, embolization, and multimodal treatment. Treatment selection and timing are based on arteriovenous malformation (AVM) features including size, location in eloquent versus noneloquent parenchyma, pattern of venous drainage, surgical access, rupture status, and previous treatments. Spetzler-Martin grading is the most commonly used classification system used to select treatment, with grades I and II lesions amenable to surgical resection alone, grade III lesions typically treated via a multimodal approach entailing preoperative embolization followed by microsurgical resection, and grades IV and V lesions generally observed unless ruptured. Embolization in the treatment of AVMs is thus most commonly used as a preoperative or, occasionally, preradiosurgical adjunct. The concept of curative AVM embolization is an attractive one that has emerged within the past few decades, with increasing clinical evidence for its safety and efficacy in recent years. Obliteration rates for curative AVM embolization will be improved by innovation in endovascular techniques and technologies.


Gestational diabetes is characterized by reduced mitochondrial protein expression and altered calcium signaling proteins in skeletal muscle.
The rising prevalence of gestational diabetes mellitus (GDM) affects up to 18% of pregnant women with immediate and long-term metabolic consequences for both mother and infant. Abnormal glucose uptake and lipid oxidation are hallmark features of GDM prompting us to use an exploratory proteomics approach to investigate the cellular mechanisms underlying differences in skeletal muscle metabolism between obese pregnant women with GDM (OGDM) and obese pregnant women with normal glucose tolerance (ONGT). Functional validation was performed in a second cohort of obese OGDM and ONGT pregnant women. Quantitative proteomic analysis in rectus abdominus skeletal muscle tissue collected at delivery revealed reduced protein content of mitochondrial complex I (C-I) subunits (NDUFS3, NDUFV2) and altered content of proteins involved in calcium homeostasis/signaling (calcineurin A, α1-syntrophin, annexin A4) in OGDM (n = 6) vs. ONGT (n = 6). Follow-up analyses showed reduced enzymatic activity of mitochondrial complexes C-I, C-III, and C-IV (-60-75%) in the OGDM (n = 8) compared with ONGT (n = 10) subjects, though no differences were observed for mitochondrial complex protein content. Upstream regulators of mitochondrial biogenesis and oxidative phosphorylation were not different between groups. However, AMPK phosphorylation was dramatically reduced by 75% in the OGDM women. These data suggest that GDM is associated with reduced skeletal muscle oxidative phosphorylation and disordered calcium homeostasis. These relationships deserve further attention as they may represent novel risk factors for development of GDM and may have implications on the effectiveness of physical activity interventions on both treatment strategies for GDM and for prevention of type 2 diabetes postpartum.


Gene fusions in tumourigenesis with particular reference to ovarian cancer.
Gene fusion, a genomic event that generates a novel gene from two independent genes, has long been known to be implicated in tumourigenesis and cancer progression. It has thus served as a diagnostic and prognostic biomarker in cancer, as well as an ideal therapeutic target in cancer therapy. Gene fusion can arise from chromosomal rearrangement and alternative splicing of transcripts, resulting in deregulation of proto-oncogenes or creation of an oncogenic novel gene. Largely facilitated by next generation sequencing technologies, a plethora of novel gene fusions have been identified in a variety of cancers, which leaves us the challenge of functionally characterising these candidate gene fusions. In this review, we summarise the molecular mechanisms, the oncogenic consequences and the therapeutic implications of verified gene fusions. We also discuss recent studies on gene fusions in both common and rare subtypes of ovarian tumours and how these findings can be translated to cancer therapies to benefit patients carrying these gene fusions.


Phylogenetic relationships among amphisbaenian reptiles based on complete mitochondrial genomic sequences.
Complete mitochondrial genomic sequences are reported from 12 members in the four families of the reptile group Amphisbaenia. Analysis of 11,946 aligned nucleotide positions (5797 informative) produces a robust phylogenetic hypothesis. The family Rhineuridae is basal and Bipedidae is the sister taxon to the Amphisbaenidae plus Trogonophidae. Amphisbaenian reptiles are surprisingly old, predating the breakup of Pangaea 200 million years before present, because successive basal taxa (Rhineuridae and Bipedidae) are situated in tectonic regions of Laurasia and nested taxa (Amphisbaenidae and Trogonophidae) are found in Gondwanan regions. Thorough sampling within the Bipedidae shows that it is not tectonic movement of Baja California away from the Mexican mainland that is primary in isolating Bipes species, but rather that primary vicariance occurred between northern and southern groups. Amphisbaenian families show parallel reduction in number of limbs and Bipes species exhibit parallel reduction in number of digits. A measure is developed for comparing the phylogenetic information content of various genes. A synapomorphic trait defining the Bipedidae is a shift from the typical vertebrate mitochondrial gene arrangement to the derived state of trnE and nad6. In addition, a tandem duplication of trnT and trnP is observed in Bipes biporus with a pattern of pseudogene formation that varies among populations. The first case of convergent rearrangement of the mitochondrial genome among animals demonstrated by complete genomic sequences is reported. Relative to most vertebrates, the Rhineuridae has the block nad6, trnE switched in order with the block cob, trnT, trnP, as they are in birds.


SLC26A1 is a major determinant of sulfate homeostasis in humans.
Sulfate plays a pivotal role in numerous physiological processes in the human body, including bone and cartilage health. A role of the anion transporter SLC26A1 (Sat1) for sulfate reabsorption in the kidney is supported by the observation of hyposulfatemia and hypersulfaturia in Slc26a1-knockout mice. The impact of SLC26A1 on sulfate homeostasis in humans remains to be defined. By combining clinical genetics, functional expression assays, and population exome analysis, we identify SLC26A1 as a sulfate transporter in humans and experimentally validate several loss-of-function alleles. Whole-exome sequencing from a patient presenting with painful perichondritis, hyposulfatemia, and renal sulfate wasting revealed a homozygous mutation in SLC26A1, which has not been previously described to the best of our knowledge. Whole-exome data analysis of more than 5,000 individuals confirmed that rare, putatively damaging SCL26A1 variants were significantly associated with lower plasma sulfate at the population level. Functional expression assays confirmed a substantial reduction in sulfate transport for the SLC26A1 mutation of our patient, which we consider to be novel, as well as for the additional variants detected in the population study. In conclusion, combined evidence from 3 complementary approaches supports SLC26A1 activity as a major determinant of sulfate homeostasis in humans. In view of recent evidence linking sulfate homeostasis with back pain and intervertebral disc disorder, our study identifies SLC26A1 as a potential target for modulation of musculoskeletal health.


Sex-dependent programming effects of prenatal glucocorticoid treatment on the developing serotonin system and stress-related behaviors in adulthood.
Prenatal stress and overexposure to glucocorticoids (GC) during development may be associated with an increased susceptibility to a number of diseases in adulthood including neuropsychiatric disorders, such as depression and anxiety. In animal models, prenatal overexposure to GC results in hyper-responsiveness to stress in adulthood, and females appear to be more susceptible than males. Here, we tested the hypothesis that overexposure to GC during fetal development has sex-specific programming effects on the brain, resulting in altered behaviors in adulthood. We examined the effects of dexamethasone (DEX; a synthetic GC) during prenatal life on stress-related behaviors in adulthood and on the tryptophan hydroxylase-2 (TpH2) gene expression in the adult dorsal raphe nucleus (DRN). TpH2 is the rate-limiting enzyme for serotonin (5-HT) synthesis and has been implicated in the etiology of human affective disorders. Timed-pregnant rats were treated with DEX from gestational days 18-22. Male and female offspring were sacrificed on the day of birth (postnatal day 0; P0), P7, and in adulthood (P80-84) and brains were examined for changes in TpH2 mRNA expression. Adult animals were also tested for anxiety- and depressive- like behaviors. In adulthood, prenatal DEX increased anxiety- and depressive- like behaviors selectively in females, as measured by decreased time spent in the center of the open field and increased time spent immobile in the forced swim test, respectively. Prenatal DEX increased TpH2 mRNA selectively in the female caudal DRN at P7, whereas it decreased TpH2 mRNA selectively in the female caudal DRN in adulthood. In animals challenged with restraint stress in adulthood, TpH2 mRNA was significantly lower in rostral DRN of prenatal DEX-treated females compared to vehicle-treated females. These data demonstrated that prenatal overexposure to GC alters the development of TpH2 gene expression and these alterations correlated with lasting behavioral changes found in adult female offspring.


Conquering the intolerable burden of malaria: what's new, what's needed: a summary.
Each year, up to three million deaths due to malaria and close to five billion episodes of clinical illness possibly meriting antimalarial therapy occur throughout the world, with Africa having more than 90% of this burden. Almost 3% of disability adjusted life years are due to malaria mortality globally, 10% in Africa. New information is presented in this supplement on malaria-related perinatal mortality, occurrence of human immunodeficiency virus in pregnancy, undernutrition, and neurologic, cognitive, and developmental sequelae. The entomologic determinants of transmission and uses of modeling for program planning and disease prediction and prevention are discussed. New data are presented from the Democratic Republic of the Congo, Tanzania, Ethiopia, and Zimbabwe on the increasing urban malaria problem and on epidemic malaria. Between 6% and 28% of the malaria burden may occur in cities, which comprise less than 2% of the African surface. Macroeconomic projections show that the costs are far greater than the costs of individual cases, with a substantial deleterious impact of malaria on schooling of patients, external investments into endemic countries, and tourism. Poor populations are at greatest risk; 58% of the cases occur in the poorest 20% of the world's population and these patients receive the worst care and have catastrophic economic consequences from their illness. This social vulnerability requires better understanding for improving deployment, access, quality, and use of effective interventions. Studies from Ghana and elsewhere indicate that for every patient with febrile illness assumed to be malaria seen in health facilities, 4-5 episodes occur in the community. Effective actions for malaria control mandate rational public policies; market forces, which often drive sales and use of drugs and other interventions, are unlikely to guarantee their use. Artemisinin-based combination therapy (ACT) for malaria is rapidly gaining acceptance as an effective approach for countering the spread and intensity of Plasmodium falciparum resistance to chloroquine, sulfadoxine/pyrimethamine, and other antimalarial drugs. Although costly, ACT ($1.20-2.50 per adult treatment) becomes more cost-effective as resistance to alternative drugs increases; early use of ACT may delay development of resistance to these drugs and prevent the medical toll associated with use of ineffective drugs. The burden of malaria in one district in Tanzania has not decreased since the primary health care approach replaced the vertical malaria control efforts of the 1960s. Despite decentralization, this situation resulted, in part, from weak district management capacity, poor coordination, inadequate monitoring, and lack of training of key staff. Experience in the Solomon Islands showed that spraying with DDT, use of insecticide-treated bed nets (ITNs), and health education were all associated with disease reduction. The use of nets permitted a reduction in DDT spraying, but could not replace it without an increased malaria incidence. Baseline data and reliable monitoring of key outcome indicators are needed to measure whether the ambitious goals for the control of malaria and other diseases has occurred. Such systems are being used for evidence-based decision making in Tanzania and several other countries. Baseline cluster sampling surveys in several countries across Africa indicate that only 53% of the children with febrile illness in malarious areas are being treated; chloroquine (CQ) is used 84% of the time, even where the drug may be ineffective. Insecticide-treated bed nets were used only 2% of the time by children less than five years of age. Progress in malaria vaccine research has been substantial over the past five years; 35 candidate malaria vaccines are in development, many of which are in clinical trials. Development of new vaccines and drugs has been the result of increased investments and formation of public-private partnerships. Before malaria vaccine becomes deployed, consideration must be given to disease burden, cost-effectiveness, financing, delivery systems, and approval by regulatory agencies. Key to evaluation of vaccine effectiveness will be collection and prompt analysis of epidemiologic information. Training of persons in every aspect of malaria research and control is essential for programs to succeed. The Multilateral Initiative on Malaria (MIM) is actively promoting research capacity strengthening and has established networks of institutions and scientists throughout the African continent, most of whom are now linked by modern information-sharing networks. Evidence over the past century is that successful control malaria programs have been linked to strong research activities. To ensure effective coordination and cooperation between the growing number of research and control coalitions forming in support of malaria activities, an umbrella group is needed. With continued support for scientists and control workers globally, particularly in low-income malarious countries, the long-deferred dream of malaria elimination can become a reality.


The plant short-chain dehydrogenase (SDR) superfamily: genome-wide inventory and diversification patterns.
Short-chain dehydrogenases/reductases (SDRs) form one of the largest and oldest NAD(P)(H) dependent oxidoreductase families. Despite a conserved 'Rossmann-fold' structure, members of the SDR superfamily exhibit low sequence similarities, which constituted a bottleneck in terms of identification. Recent classification methods, relying on hidden-Markov models (HMMs), improved identification and enabled the construction of a nomenclature. However, functional annotations of plant SDRs remain scarce. Wide-scale analyses were performed on ten plant genomes. The combination of hidden Markov model (HMM) based analyses and similarity searches led to the construction of an exhaustive inventory of plant SDR. With 68 to 315 members found in each analysed genome, the inventory confirmed the over-representation of SDRs in plants compared to animals, fungi and prokaryotes. The plant SDRs were first classified into three major types - 'classical', 'extended' and 'divergent' - but a minority (10% of the predicted SDRs) could not be classified into these general types ('unknown' or 'atypical' types). In a second step, we could categorize the vast majority of land plant SDRs into a set of 49 families. Out of these 49 families, 35 appeared early during evolution since they are commonly found through all the Green Lineage. Yet, some SDR families - tropinone reductase-like proteins (SDR65C), 'ABA2-like'-NAD dehydrogenase (SDR110C), 'salutaridine/menthone-reductase-like' proteins (SDR114C), 'dihydroflavonol 4-reductase'-like proteins (SDR108E) and 'isoflavone-reductase-like' (SDR460A) proteins - have undergone significant functional diversification within vascular plants since they diverged from Bryophytes. Interestingly, these diversified families are either involved in the secondary metabolism routes (terpenoids, alkaloids, phenolics) or participate in developmental processes (hormone biosynthesis or catabolism, flower development), in opposition to SDR families involved in primary metabolism which are poorly diversified. The application of HMMs to plant genomes enabled us to identify 49 families that encompass all Angiosperms ('higher plants') SDRs, each family being sufficiently conserved to enable simpler analyses based only on overall sequence similarity. The multiplicity of SDRs in plant kingdom is mainly explained by the diversification of large families involved in different secondary metabolism pathways, suggesting that the chemical diversification that accompanied the emergence of vascular plants acted as a driving force for SDR evolution.


A Rapid SARS-CoV-2 RT-PCR Assay for Low Resource Settings.
Quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay is the gold standard recommended to test for acute SARS-CoV-2 infection. However, it generally requires expensive equipment such as RNA isolation instruments and real-time PCR thermal cyclers. As a pandemic, COVID-19 has spread indiscriminately, and many low resource settings and developing countries do not have the means for fast and accurate COVID-19 detection to control the outbreak. Additionally, long assay times, in part caused by slow sample preparation steps, have created a large backlog when testing patient samples suspected of COVID-19. With many PCR-based molecular assays including an extraction step, this can take a significant amount of time and labor, especially if the extraction is performed manually. Using COVID-19 clinical specimens, we have collected evidence that the RT-qPCR assay can feasibly be performed directly on patient sample material in virus transport medium (VTM) without an RNA extraction step, while still producing sensitive test results. If RNA extraction steps can be omitted without significantly affecting clinical sensitivity, the turn-around time of COVID-19 tests, and the backlog we currently experience can be reduced drastically. Furthermore, our data suggest that rapid RT-PCR can be implemented for sensitive and specific molecular diagnosis of COVID-19 in locations where sophisticated laboratory instruments are not available. Our USD 300 set up achieved rapid RT-PCR using thin-walled PCR tubes and a water bath setup using sous vide immersion heaters, a Raspberry Pi computer, and a single servo motor that can process up to 96 samples at a time. Using COVID-19 positive clinical specimens, we demonstrated that RT-PCR assays can be performed in as little as 12 min using untreated samples, heat-inactivated samples, or extracted RNA templates with our low-cost water bath setup. These findings can help rapid COVID-19 testing to become more accessible and attainable across the globe.


Hereditary Angioedema Attacks: Local Swelling at Multiple Sites.
Hereditary angioedema (HAE) patients experience recurrent local swelling in various parts of the body including painful swelling of the intestine and life-threatening laryngeal oedema. Most HAE literature is about attacks located in one anatomical site, though it is mentioned that HAE attacks may also involve multiple anatomical sites simultaneously. A detailed description of such multi-location attacks is currently lacking. This study investigated the occurrence, severity and clinical course of HAE attacks with multiple anatomical locations. HAE patients included in a clinical database of recombinant human C1-inhibitor (rhC1INH) studies were evaluated. Visual analog scale scores filled out by the patients for various symptoms at various locations and investigator symptoms scores during the attack were analysed. Data of 219 eligible attacks in 119 patients was analysed. Thirty-three patients (28%) had symptoms at multiple locations in anatomically unrelated regions at the same time during their first attack. Up to five simultaneously affected locations were reported. The observation that severe HAE attacks often affect multiple sites in the body suggests that HAE symptoms result from a systemic rather than from a local process as is currently believed.


Clinical features of bowel anisakiasis in Japan.
Bowel anisakiasis is rare, and the incidence and clinical features of this condition remain unclear. Using the Japanese Diagnosis Procedure Combination (DPC) in-patient database, we identified 201 cases of bowel anisakiasis between the months of July and December during 2007 and 2008. More than 70% were males. The average age was 54.5 years. Overall, 102 (50.7%) cases had ileus, 16 (8.0%) had perforation or peritonitis, and 4 (2.0%) had intestinal bleeding. Allergic responses, including urticaria, were found in seven (3.5%) patients. Fourteen (7.0%) cases underwent open surgery. Three (1.5%) underwent colonoscopic removal of Anisakis larvae. The average length of stay in the hospital was 9.6 days. The annual incidence of bowel anisakiasis is estimated to be about 3.0 per 1 million people per year. It is important to continue collecting all available data to monitor the trends of this distressing condition.


Agreeableness and activity engagement in nursing home residents with dementia.
Residents with dementia are the least likely to be engaged in the nursing home and often spend most of their time doing nothing at all. However, resident participation in meaningful activities is important to promote both physical and psychological health. Tailoring activities to individual functional abilities and personality preferences improves both the time and level of participation. This pilot study used an analysis of covariance procedure to test the relationship between the personality trait of agreeableness and engagement when activities are ideally tailored to ability and interest. No significant difference was found between the high and low agreeableness groups, indicating that residents were more engaged when activities were individually tailored, regardless of their agreeableness level. Although low agreeableness may pose a challenge when implementing activities for people with dementia, the results of this study suggest that tailoring activities to functional ability and interest may overcome the effects.


Safety of Low-Dose Oral Minoxidil in Patients With Hypertension and Arrhythmia: A Multicenter Study of 264 Patients.
Systemic adverse effects (AE) are a major concern of low-dose oral minoxidil (LDOM) treatment, especially in patients with arterial hypertension or arrhythmia. The objective of this study was to evaluate the safety of LDOM in patients with hypertension or arrhythmia. Retrospective multicenter study of patients with hypertension or arrhythmia treated with LDOM for any type of alopecia. A total of 254 patients with hypertension [176 women (69.3%) and 78 men (30.7%)] with a mean age of 56.9 years (range 19-82) were included. From them, the dose of LDOM was titrated in 128 patients, allowing the analysis of 382 doses. Patients were receiving a mean of 1.45 (range 0-5) antihypertensive drugs. Systemic AE were detected in 26 cases (6.8%) and included lightheadedness (3.1%), fluid retention (2.6%), general malaise (0.8%), tachycardia (0.8%) and headache (0.5%), leading to LDOM discontinuation in 6 cases (1.5%). Prior treatment with doxazosin (P<0.001), or with three or more antihypertensive drugs (P=0.012) was associated with a higher risk of discontinuation of LDOM. LDOM treatment showed a favorable safety profile in patients with hypertension or arrhythmia, similar to general population.


Functional, molecular and proteomic characterisation of bone marrow mesenchymal stem cells in rheumatoid arthritis.
Bone marrow (BM) mesenchymal stem cells (MSCs) are being considered as potential therapeutic agents in various inflammatory autoimmune diseases for their tissue-repair and anti-inflammatory tissue-protective properties. This study investigates the reserves and function, the molecular and proteomic profile and the differentiation potential of BM MSCs in patients with active rheumatoid arthritis (RA). We evaluated the frequency of MSCs in the BM mononuclear cell fraction using a limiting dilution assay, the proliferative/clonogenic potential and the capacity of cells to differentiate towards the osteogenic/chondrogenic/adipogenic lineages using appropriate culture conditions. We also assessed the molecular and proteomic characteristics in terms of inflammatory cytokine gene and protein expression, the relative telomere length and the survival characteristics of BM MSCs. MSCs from patients with RA (n = 26) and age- and sex-matched healthy individuals (n = 21) were similar in frequency, differentiation potential, survival, immunophenotypic characteristics, and protein profile. Patient MSCs, however, had impaired clonogenic and proliferative potential in association with premature telomere length loss. Transcriptome analysis revealed differential expression of genes related to cell adhesion processes and cell cycle progression beyond the G1 phase. Previous treatment with methotrexate, corticosteroids, anti-cytokine and biological agents or other disease-modifying anti-inflammatory drugs did not correlate with the clonogenic and proliferative impairment of BM MSCs. In spite of some restrictions related to the impaired clonogenic and proliferative potential, our findings support the use of autologous BM MSCs in RA and may have important implications for the ongoing efforts to repair tissue injury commonly seen in the course of the disease.


Magnesium protects against cisplatin-induced acute kidney injury without compromising cisplatin-mediated killing of an ovarian tumor xenograft in mice.
Cisplatin, a commonly used chemotherapeutic for ovarian and other cancers, leads to hypomagnesemia in most patients and causes acute kidney injury (AKI) in 25-30% of patients. Previously, we showed that magnesium deficiency worsens cisplatin-induced AKI and magnesium replacement during cisplatin treatment protects against cisplatin-mediated AKI in non-tumor-bearing mice (Solanki MH, Chatterjee PK, Gupta M, Xue X, Plagov A, Metz MH, Mintz R, Singhal PC, Metz CN. Am J Physiol Renal Physiol 307: F369-F384, 2014). This study investigates the role of magnesium in cisplatin-induced AKI using a human ovarian tumor (A2780) xenograft model in mice and the effect of magnesium status on tumor growth and the chemotherapeutic efficacy of cisplatin in vivo. Tumor progression was unaffected by magnesium status in saline-treated mice. Cisplatin treatment reduced tumor growth in all mice, irrespective of magnesium status. In fact, cisplatin-treated magnesium-supplemented mice had reduced tumor growth after 3 wk compared with cisplatin-treated controls. While magnesium status did not interfere with tumor killing by cisplatin, it significantly affected renal function following cisplatin. Cisplatin-induced AKI was enhanced by magnesium deficiency, as evidenced by increased blood urea nitrogen, creatinine, and other markers of renal damage. This was accompanied by reduced renal mRNA expression of the cisplatin efflux transporter Abcc6. These effects were significantly reversed by magnesium replacement. On the contrary, magnesium status did not affect the mRNA expression of cisplatin uptake or efflux transporters by the tumors in vivo. Finally, magnesium deficiency enhanced platinum accumulation in the kidneys and renal epithelial cells, but not in the A2780 tumor cells. These findings demonstrate the renoprotective role of magnesium during cisplatin AKI, without compromising the chemotherapeutic efficacy of cisplatin in an ovarian tumor-bearing mouse model.


Androgenic responses to resistance exercise: effects of feeding and L-carnitine.
The purpose of this investigation was to determine the effects of 3 wk of L-carnitine L-tartrate (LCLT) supplementation and post-resistance-exercise (RE) feeding on hormonal and androgen receptor (AR) responses. Ten resistance-trained men (mean+/-SD: age, 22+/-1 yr; mass, 86.3+/-15.3 kg; height, 181+/-11 cm) supplemented with LCLT (equivalent to 2 g of L-carnitine per day) or placebo (PL) for 21 d, provided muscle biopsies for AR determinations, then performed two RE protocols: one followed by water intake, and one followed by feeding (8 kcal.kg body mass, consisting of 56% carbohydrate, 16% protein, and 28% fat). RE protocols were randomized and included serial blood draws and a 1-h post-RE biopsy. After a 7-d washout period, subjects crossed over, and all experimental procedures were repeated. LCLT supplementation upregulated (P<0.05) preexercise AR content compared with PL (12.9+/-5.9 vs 11.2+/-4.0 au, respectively). RE increased (P<0.05) AR content compared with pre-RE values in the PL trial only. Post-RE feeding significantly increased AR content compared with baseline and water trials for both LCLT and PL. Serum total testosterone concentrations were suppressed (P<0.05) during feeding trials with respect to corresponding water and pre-RE values. Luteinizing hormone demonstrated subtle, yet significant changes in response to feeding and LCLT. In summary, these data demonstrated that: 1) feeding after RE increased AR content, which may result in increased testosterone uptake, and thus enhanced luteinizing hormone secretion via feedback mechanisms; and 2) LCLT supplementation upregulated AR content, which may promote recovery from RE.


Drug resistance in Salmonella strains isolated from domestic wastewater before and after treatment in stabilization ponds in an arid region (Marrakech, Morocco).
Some 118 Salmonella strains isolated before and after treatment in stabilization ponds were tested for antimicrobial resistance. In the treatment plant, which decreases the abundance of Salmonella by 99%, a significantly lower level of antibiotic resistance (P<0.01) was identified at the system's inflow point (19%) than at its outflow (29%). The serotypes most frequently identified as having multiple antibiotic resistance were Salmonella paratyphi B and S. typhimurium. High tetracycline resistance was observed at all sampling points, followed by resistance to ampicillin and streptomycin. Antibiotic resistance can be transferred from Salmonella to other members of the Enterobacteriaceae family, such as Escherichia coli K12; transfer frequencies in nutrient broth and filtered sewage water were 4.5×10(-4) and 7×10(-7), respectively.


Population structure in the Mediterranean basin: a Y chromosome perspective.
The Mediterranean region has been characterised by a number of pre-historical and historical demographic events whose legacy on the current genetic landscape is still a matter of debate. In order to investigate the degree of population structure across the Mediterranean, we have investigated Y chromosome variation in a large dataset of Mediterranean populations, 11 of which are first described here. Our analyses identify four main clusters in the Mediterranean that can be labelled as North Africa, Arab, Central-East and West Mediterranean. In particular, Near Eastern samples tend to separate according to the presence of Arab Y chromosome lineages, suggesting that the Arab expansion played a major role in shaping the current genetic structuring within the Fertile Crescent.


Therapeutic selective neck dissection outcomes.
To evaluate the effectiveness of selective neck dissection in patients with nodal metastases from head and neck squamous cell carcinoma. Historical cohort study. Academic medical center. A chart review was performed on 156 subjects with clinically positive regional nodal metastases managed initially with surgery, including neck dissection. Sixty-nine subjects underwent selective neck dissection (less than 5 levels), and the majority received postoperative radiotherapy (80%). Primary outcomes included Kaplan-Meier three-year ipsilateral regional control and five-year overall survival. Cox proportional univariate and multivariate analyses were performed to determine those factors associated with outcome. There were two ipsilateral regional recurrences among those undergoing selective neck dissection, yielding a regional control rate of 95.9 percent. Among those undergoing comprehensive neck dissection, nine ipsilateral regional recurrences occurred, yielding a control rate of 86.0 percent (P = 0.053). No selective neck dissection recurrences occurred in a preserved level. Selective neck dissection, as compared to comprehensive neck dissection, was not adversely associated with regional recurrence, survival, or distant metastasis, even after adjusting for possible confounders (hazard ratio 0.21, P = 0.055). These results demonstrate high rates of regional disease control (96%) following selective neck dissection and radiotherapy in patients with positive neck node metastases. In this population, performing selective neck dissection with adjuvant radiotherapy for the majority of patients is supported as an effective treatment approach.


Application of Bayesian logistic regression to mining biomedical data.
Mining high dimensional biomedical data with existing classifiers is challenging and the predictions are often inaccurate. We investigated the use of Bayesian Logistic Regression (B-LR) for mining such data to predict and classify various disease conditions. The analysis was done on twelve biomedical datasets with binary class variables and the performance of B-LR was compared to those from other popular classifiers on these datasets with 10-fold cross validation using the WEKA data mining toolkit. The statistical significance of the results was analyzed by paired two tailed t-tests and non-parametric Wilcoxon signed-rank tests. We observed overall that B-LR with non-informative Gaussian priors performed on par with other classifiers in terms of accuracy, balanced accuracy and AUC. These results suggest that it is worthwhile to explore the application of B-LR to predictive modeling tasks in bioinformatics using informative biological prior probabilities. With informative prior probabilities, we conjecture that the performance of B-LR will improve.


Identification and expression analysis of an atypical chemokine receptor-2 (ACKR2)/CC chemokine binding protein-2 (CCBP2) in rainbow trout (Oncorhynchus mykiss).
Atypical chemokine receptors (ACKRs) have emerged as key components of the chemokine system, with an essential regulatory function in innate and adaptive immune responses and inflammation. In mammals ACKR2 is a 'scavenging' receptor for inflammatory CC chemokines and plays a central role in the resolution of in vivo inflammatory responses. An ACKR2 like gene has been identified and cloned in rainbow trout (Teleostei) in the present study, enabling the further identification of this molecule in another group of ray-finned teleost fish (Holostei), in a lobe-finned fish (Sarcopterygii-coelacanth), and in reptiles. The identity of these ACKR2 molecules is supported by their conserved structure, and by phylogenetic tree and synteny analysis. Trout ACKR2 is highly expressed in spleen and head kidney, suggesting a homeostatic role of this receptor in limiting the availability of its potential ligands. Trout ACKR2 expression can be modulated in vivo by bacterial and parasitic infections, and in vitro by PAMPs (poly I:C and peptidoglycan) and cytokines (IL-6, TNF-α, IFN-γ and IL-21) in a time dependent manner. These patterns of expression and modulation suggest that trout ACKR2 is regulated in a complex way and has an important role in control of the chemokine network in fish as in mammals.


An investigation of institutional characteristics associated with response rates in mail surveys of community hospitals.
This paper describes the nature and strength of the relationship between six institutional characteristics of U.S. community hospitals and the rates of response of these hospitals to a nationwide survey conducted by the American Hospital Association (AHA). Furthermore, it demonstrates how one can calculate accurately the relative probability of response of hospitals with various combinations of these characteristics. The six characteristics studied were bed size, teaching status, AHA membership status, location within or without a Standard Metropolitan Statistical Area, investor or other form of ownership, and control by State or local government or by another type of organization.The six characteristics were treated as dichotomous variables throughout most of the analysis. Odds ratios were calculated for each variable as a preliminary measure of the strength of its association with response.The effects of confounding on those odds ratios were controlled for by multiple logistic regression, which estimates the probability of response of hospitals with given characteristics. A logistic odds ratio was calculated for each variable to estimate the independent effect that specified values had on the odds of responding. All variables except status as a teaching or nonteaching hospital were shown to have a significant relationship to response.Hospitals were divided into classes according to probability of response, and the probability of response was compared with actual rates of response in the survey. Both increase at equal rates. Similar results were obtained by applying this method to two other, dissimilar, surveys.This method is generalizable to other surveys and should prove useful to researchers wishing to increase response rates in their surveys of hospitals.


Use of a low-cost, commercially available gaming console (Wii) for rehabilitation of an adolescent with cerebral palsy.
The purpose of this retrospective and prospective case report is to describe the feasibility and outcomes of using a low-cost, commercially available gaming system (Wii) to augment the rehabilitation of an adolescent with cerebral palsy. The patient was an adolescent with spastic diplegic cerebral palsy classified as GMFCS level III who was treated during a summer session in a school-based setting. The patient participated in 11 training sessions, 2 of which included other players. Sessions were between 60 and 90 minutes in duration. Training was performed using the Wii sports games software, including boxing, tennis, bowling, and golf. He trained in both standing and sitting positions. Three main outcome measures were used: (1) visual-perceptual processing, using a motor-free perceptual test (Test of Visual Perceptual Skills, third edition); (2) postural control, using weight distribution and sway measures; and (3) functional mobility, using gait distance. Improvements in visual-perceptual processing, postural control, and functional mobility were measured after training. The feasibility of using the system in the school-based setting during the summer session was supported. For this patient whose rehabilitation was augmented with the Wii, there were positive outcomes at the impairment and functional levels. Multiple hypotheses were proposed for the findings that may be the springboard for additional research. To the authors' knowledge, this is the first published report on using this particular low-cost, commercially available gaming technology for rehabilitation of a person with cerebral palsy.


Prevalence of depression among people with type 2 diabetes mellitus: a cross sectional study in Palestine.
Diabetes mellitus is a common chronic metabolic disorder and one of the main causes of death in Palestine. Palestinians are continuously living under stressful economic and military conditions which make them psychologically vulnerable. The purpose of this study was to investigate the prevalence of depression among type II diabetic patients and to examine the relationship between depression and socio-demographic factors, clinical factors, and glycemic control. This was a cross-sectional study at Al-Makhfiah primary healthcare center, Nablus, Palestine. Two hundred and ninety-four patients were surveyed for the presence of depressive symptoms using Beck Depression Inventory (BDI-II) scale. Patients' records were reviewed to obtain data pertaining to age, sex, marital status, Body Mass Index (BMI), level of education, smoking status, duration of diabetes mellitus, glycemic control using HbA1C test, use of insulin, and presence of additional illnesses. Patients' medication adherence was assessed using the 8-item Morisky Medication Adherence Scale (MMAS-8). One hundred and sixty four patients (55.8%) of the total sample were females and 216 (73.5%) were < 65 years old. One hundred and twenty patients (40.2%) scored ≥ 16 on BDI-II scale. Statistical significant association was found between high BDI-II score (≥ 16) and female gender, low educational level, having no current job, having multiple additional illnesses, low medication adherence and obesity (BMI ≥ 30 kg/m2). No significant association between BDI score and glycemic control, duration of diabetes, and other socio-demographic factors was found. Multivatriate analysis showed that low educational level, having no current job, having multiple additional illnesses and low medication adherence were significantly associated with high BDI-II scores. Prevalence of depression found in our study was higher than that reported in other countries. Although 40% of the screened patients were potential cases of depression, none were being treated with anti-depressants. Psychosocial assessment should be part of routine clinical evaluation of these patients at primary healthcare clinics to improve quality of life and decrease adverse outcomes among diabetic patients.


Prognosis of fracture: evaluation of predictive accuracy of the FRAX algorithm and Garvan nomogram.
We evaluated the prognostic accuracy of fracture risk assessment tool (FRAX) and Garvan algorithms in an independent Australian cohort. The results suggest comparable performance in women but relatively poor fracture risk discrimination in men by FRAX. These data emphasize the importance of external validation before widespread clinical implementation of prognostic tools in different cohorts. Absolute risk assessment is now recognized as a preferred approach to guide treatment decision. The present study sought to evaluate accuracy of the FRAX and Garvan algorithms for predicting absolute risk of osteoporotic fracture (hip, spine, humerus, or wrist), defined as major in FRAX, in a clinical setting in Australia. A retrospective validation study was conducted in 144 women (69 fractures and 75 controls) and 56 men (31 fractures and 25 controls) aged between 60 and 90 years. Relevant clinical data prior to fracture event were ascertained. Based on these variables, predicted 10-year probabilities of major fracture were calculated from the Garvan and FRAX algorithms, using US (FRAX-US) and UK databases (FRAX-UK). Area under the receiver operating characteristic curves (AUC) was computed for each model. In women, the average 10-year probability of major fracture was consistently higher in the fracture than in the nonfracture group: Garvan (0.33 vs. 0.15), FRAX-US (0.30 vs. 0.19), and FRAX-UK (0.17 vs. 0.10). In men, although the Garvan model yielded higher average probability of major fracture in the fracture group (0.32 vs. 0.14), the FRAX algorithm did not: FRAX-US (0.17 vs. 0.19) and FRAX-UK (0.09 vs. 0.12). In women, AUC for the Garvan, FRAX-US, and FRAX-UK algorithms were 0.84, 0.77, and 0.78, respectively, vs. 0.76, 0.54, and 0.57, respectively, in men. In this analysis, although both approaches were reasonably accurate in women, FRAX discriminated fracture risk poorly in men. These data support the concept that all algorithms need external validation before clinical implementation.


The NATO project: nanoparticle-based countermeasures for microgravity-induced osteoporosis.
Recent advances in nanotechnology applied to medicine and regenerative medicine have an enormous and unexploited potential for future space and terrestrial medical applications. The Nanoparticles and Osteoporosis (NATO) project aimed to develop innovative countermeasures for secondary osteoporosis affecting astronauts after prolonged periods in space microgravity. Calcium- and Strontium-containing hydroxyapatite nanoparticles (nCa-HAP and nSr-HAP, respectively) were previously developed and chemically characterized. This study constitutes the first investigation of the effect of the exogenous addition of nCa-HAP and nSr-HAP on bone remodeling in gravity (1 g), Random Positioning Machine (RPM) and onboard International Space Station (ISS) using human bone marrow mesenchymal stem cells (hBMMSCs). In 1 g conditions, nSr-HAP accelerated and improved the commitment of cells to differentiate towards osteoblasts, as shown by the augmented alkaline phosphatase (ALP) activity and the up-regulation of the expression of bone marker genes, supporting the increased extracellular bone matrix deposition and mineralization. The nSr-HAP treatment exerted a protective effect on the microgravity-induced reduction of ALP activity in RPM samples, and a promoting effect on the deposition of hydroxyapatite crystals in either ISS or 1 g samples. The results indicate the exogenous addition of nSr-HAP could be potentially used to deliver Sr to bone tissue and promote its regeneration, as component of bone substitute synthetic materials and additive for pharmaceutical preparation or food supplementary for systemic distribution.


Phylogenetic MCMC algorithms are misleading on mixtures of trees.
Markov chain Monte Carlo (MCMC) algorithms play a critical role in the Bayesian approach to phylogenetic inference. We present a theoretical analysis of the rate of convergence of many of the widely used Markov chains. For N characters generated from a uniform mixture of two trees, we prove that the Markov chains take an exponentially long (in N) number of iterations to converge to the posterior distribution. Nevertheless, the likelihood plots for sample runs of the Markov chains deceivingly suggest that the chains converge rapidly to a unique tree. Our results rely on novel mathematical understanding of the log-likelihood function on the space of phylogenetic trees. The practical implications of our work are that Bayesian MCMC methods can be misleading when the data are generated from a mixture of trees. Thus, in cases of data containing potentially conflicting phylogenetic signals, phylogenetic reconstruction should be performed separately on each signal.


One-year clinical evaluation of compomer restorations placed in general practice.
This study evaluated the in-service performance, after 1 year, of a sample of F2000 compomer restorations placed by 10 general dental practitioners as part of handling evaluation of this material. Five general dental practitioners who participated in the handling evaluation were asked to cooperate in the 1-year assessment of the restorations. The practitioners were requested to contact patients and inquire if they were willing to attend at a specified time for examination of their restorations by a trained evaluator who would travel to the various practice locations. A total of 84 restorations were presented for examination. The mean age of the restorations at examination was 14 months. The restorations were assessed for anatomic form, marginal adaptation, surface roughness, sensitivity or discomfort, gingival condition, and presence or absence of secondary caries. Eighty-two restorations were intact. No secondary caries was detected. Optimal scores were obtained in a high proportion of the assessments. All patients reported satisfaction with the restorations, both in appearance and surface texture, and no symptoms were reported. The F2000 compomer restorations, placed in conjunction with its bonding system in general practice conditions, were found to be performing satisfactorily after 1 year.


Psychosocial Well-Being of Young People Who Participated in a Support Group Following the Loss of a Parent to Cancer.
Despite the evidence of unmet support needs among young people who have lost a parent to cancer, only a few support group initiatives have been reported. This observational prospective study explored the psychosocial well-being of young people who participated in support groups at a Swedish specialist palliative care setting. On three occasions, 29 participants, aged 16-28 years, answered questionnaires covering characteristics of the participants, circumstances of the losses, psychosocial well-being of the young people, and their own assessment of the support groups. The support groups attracted mostly young women who were often unprepared for the loss. The living arrangements differed between younger and older participants; however, the loss-related variables did not differ. Significant positive changes were found regarding a sense of meaning in their future life and life satisfaction. The helpfulness of the group was assessed as high/very high and the group brought a valuable fellowship with others in a similar situation. Universality and beneficial interactions were reported and strengthened psychosocial well-being developed over time. This change, according to the young people themselves, may be attributed to the group support. The findings are useful for planning interventions to support young people in bereavement in order to enhance their psychosocial well-being.


Social network of PESCA (Open Source Platform for eHealth).
Information and Communication Technologies (ICTs) are revolutionizing how healthcare systems deliver top-quality care to citizens. In this way, Open Source Software (OSS) has demonstrated to be an important strategy to spread ICTs use. Several human and technological barriers in adopting OSS for healthcare have been identified. Human barriers include user acceptance, limited support, technical skillfulness, awareness, resistance to change, etc., while Technological barriers embrace need for open standards, heterogeneous OSS developed without normalization and metrics, lack of initiatives to evaluate existing health OSS and need for quality control and functional validation. The goals of PESCA project are to create a platform of interoperable modules to evaluate, classify and validate good practices in health OSS. Furthermore, a normalization platform will provide interoperable solutions in the fields of healthcare services, health surveillance, health literature, and health education, knowledge and research. Within the platform, the first goal to achieve is the setup of the collaborative work infrastructure. The platform is being organized as a Social Network which works to evaluate five scopes of every existing open source tools for eHealth: Open Source Software, Quality, Pedagogical, Security and privacy and Internationalization/I18N. In the meantime, the knowledge collected from the networking will configure a Good Practice Repository on eHealth promoting the effective use of ICT on behalf of the citizen's health.


Lineage-specific expansions of retroviral insertions within the genomes of African great apes but not humans and orangutans.
Retroviral infections of the germline have the potential to episodically alter gene function and genome structure during the course of evolution. Horizontal transmissions between species have been proposed, but little evidence exists for such events in the human/great ape lineage of evolution. Based on analysis of finished BAC chimpanzee genome sequence, we characterize a retroviral element (Pan troglodytes endogenous retrovirus 1 [PTERV1]) that has become integrated in the germline of African great ape and Old World monkey species but is absent from humans and Asian ape genomes. We unambiguously map 287 retroviral integration sites and determine that approximately 95.8% of the insertions occur at non-orthologous regions between closely related species. Phylogenetic analysis of the endogenous retrovirus reveals that the gorilla and chimpanzee elements share a monophyletic origin with a subset of the Old World monkey retroviral elements, but that the average sequence divergence exceeds neutral expectation for a strictly nuclear inherited DNA molecule. Within the chimpanzee, there is a significant integration bias against genes, with only 14 of these insertions mapping within intronic regions. Six out of ten of these genes, for which there are expression data, show significant differences in transcript expression between human and chimpanzee. Our data are consistent with a retroviral infection that bombarded the genomes of chimpanzees and gorillas independently and concurrently, 3-4 million years ago. We speculate on the potential impact of such recent events on the evolution of humans and great apes.


Exploring a massage intervention for parents and their children with autism: the implications for bonding and attachment.
This exploratory study aimed to address two questions: (1) What does touch mean between parents and their children with autism on completion of a massage intervention? (2) Do parents feel that their relationship with their children has changed on completion of a massage intervention? Fourteen parents agreed to be interviewed. Data were collected before the massage intervention (baseline), immediately after the massage intervention and 16 weeks from baseline and were analysed using interpretative phenomenological analysis. At baseline, parents felt distressed that they felt unable to get 'close' to their children. After the intervention, parents reported feeling physically and emotionally closer to their children. Children expressed a range of cues to initiate massage at home. These benefits were maintained at follow-up for parents who continued to use massage at home. In conclusion, giving massage to children with autism may help to enhance the emotional bond between parent and child.


Genome-wide copy number analysis on DNA from fetal cells isolated from the blood of pregnant women.
Non-invasive prenatal testing (NIPT) based on fetal cells in maternal blood has the advantage over NIPT based on circulating cell-free fetal DNA in that there is no contamination with maternal DNA. This will most likely result in better detection of chromosomal aberrations including subchromosomal defects. The objective of this study was to test whether fetal cells enriched from maternal blood can be used for cell-based NIPT. We present a method for enriching fetal cells from maternal blood, subsequent amplification of the fetal genome and detection of chromosomal and subchromosomal variations in the genome. An average of 12.8 fetal cells from 30 mL of maternal blood were recovered using our method. Subsequently, whole genome amplification on fetal cells resulted in amplified fetal DNA in amounts and quality high enough to generate array comparative genomic hybridization as well as next-generation sequencing profiles. From one to two fetal cells, we were able to demonstrate copy number differences of whole chromosomes (21, X-, and Y) as well as subchromosomal aberrations (ring X). Intact fetal cells can be isolated from every maternal blood sample. Amplified DNA from isolated fetal cells enabled genetic analysis by array comparative genomic hybridization and next-generation sequencing. © 2016 John Wiley & Sons, Ltd.


Validation of an algorithm for the nonrigid registration of longitudinal breast MR images using realistic phantoms.
The authors present a method to validate coregistration of breast magnetic resonance images obtained at multiple time points during the course of treatment. In performing sequential registration of breast images, the effects of patient repositioning, as well as possible changes in tumor shape and volume, must be considered. The authors accomplish this by extending the adaptive bases algorithm (ABA) to include a tumor-volume preserving constraint in the cost function. In this study, the authors evaluate this approach using a novel validation method that simulates not only the bulk deformation associated with breast MR images obtained at different time points, but also the reduction in tumor volume typically observed as a response to neoadjuvant chemotherapy. For each of the six patients, high-resolution 3D contrast enhanced T1-weighted images were obtained before treatment, after one cycle of chemotherapy and at the conclusion of chemotherapy. To evaluate the effects of decreasing tumor size during the course of therapy, simulations were run in which the tumor in the original images was contracted by 25%, 50%, 75%, and 95%, respectively. The contracted area was then filled using texture from local healthy appearing tissue. Next, to simulate the post-treatment data, the simulated (i.e., contracted tumor) images were coregistered to the experimentally measured post-treatment images using a surface registration. By comparing the deformations generated by the constrained and unconstrained version of ABA, the authors assessed the accuracy of the registration algorithms. The authors also applied the two algorithms on experimental data to study the tumor volume changes, the value of the constraint, and the smoothness of transformations. For the six patient data sets, the average voxel shift error (mean +/- standard deviation) for the ABA with constraint was 0.45 +/- 0.37, 0.97 +/- 0.83, 1.43 +/- 0.96, and 1.80 +/- 1.17 mm for the 25%, 50%, 75%, and 95% contraction simulations, respectively. In comparison, the average voxel shift error for the unconstrained ABA was 0.46 +/- 0.29, 1.13 +/- 1.17, 2.40 +/- 2.04, and 3.53 +/- 2.89 mm, respectively. These voxel shift errors translate into compression of the tumor volume: The ABA with constraint returned volumetric errors of 2.70 +/- 4.08%, 7.31 +/- 4.52%, 9.28 +/- 5.55%, and 13.19 +/- 6.73% for the 25%, 50%, 75%, and 95% contraction simulations, respectively, whereas the unconstrained ABA returned volumetric errors of 4.00 +/- 4.46%, 9.93 +/- 4.83%, 19.78 +/- 5.657%, and 29.75 +/- 15.18%. The ABA with constraint yields a smaller mean shift error, as well as a smaller volume error (p = 0.031 25 for the 75% and 95% contractions), than the unconstrained ABA for the simulated sets. Visual and quantitative assessments on experimental data also indicate a good performance of the proposed algorithm. The ABA with constraint can successfully register breast MR images acquired at different time points with reasonable error. To the best of the authors' knowledge, this is the first report of an attempt to quantitatively assess in both phantoms and a set of patients the accuracy of a registration algorithm for this purpose.


Stress reaction of the humerus in tennis players.
Overuse injuries are a frequent occurrence among competitive athletes. When analysing the incidence of overuse injuries in tennis players, it has been determined that a significant number of these injures occur in the upper limb area. In this study, we describe five cases of a stress-induced injury to the middle and distal humerus occurring mainly due to repetitive serving. Athletes studied were competitive tennis players and elite junior players, two of whom played at international level. Four of the five were male. In all cases, diagnosis was confirmed by magnetic resonance imaging examination. The treatment of middle and distal humeral stress reactions consisted of physical therapy, which focused on analgesia and muscle strengthening. In addition, we analysed each tennis player's strokes in order to identify modifications that would decrease the amount of stress that the upper limbs were subjected to during the service motion. The players in our study missed on average 3 weeks of play and at follow-up after 1 year were able to play symptom free. Our study highlights the need for coaches, physicians and players to be aware of distal humeral pain and understand treatment options in order to prevent further injury, including stress fractures.


Neutralizing antibodies generated during natural HIV-1 infection: good news for an HIV-1 vaccine?
Most existing viral vaccines generate antibodies that either block initial infection or help eradicate the virus before it can cause disease. For HIV-1, obstacles to eliciting protective neutralizing antibodies (NAbs) have often seemed insurmountable. The target of HIV-specific NAbs, the viral envelope glycoprotein (Env), is highly variable in amino acid sequence and glycosylation pattern. Conserved elements of HIV-1 Env seem to be poorly immunogenic, and previous attempts to generate broadly reactive NAbs by vaccination have proven ineffective. However, recent studies show that antibodies in the sera of some HIV-1-infected individuals can neutralize diverse HIV-1 isolates. Detailed analyses of these sera provide new insights into the viral epitopes targeted by broadly reactive NAbs. The findings discussed here suggest that the natural NAb response to HIV-1 can inform future vaccine design. A concerted effort of structure-based vaccine design will help guide the development of improved antibody-based vaccines for HIV-1.


Pheromone receptors in mammals.
In most mammals, pheromone perception mediates intraspecies interactions related to reproduction, such as mate recognition, intermale aggressive behaviors, or exchanges between females and their offspring. Recent molecular findings, particularly the identification of two large pheromone receptor gene superfamilies, provide today invaluable tools to better understand the way mammals make sense of pheromonal information.


Histone Acetyltransferase KAT2A Stabilizes Pluripotency with Control of Transcriptional Heterogeneity.
Cell fate transitions in mammalian stem cell systems have often been associated with transcriptional heterogeneity; however, existing data have failed to establish a functional or mechanistic link between the two phenomena. Experiments in unicellular organisms support the notion that transcriptional heterogeneity can be used to facilitate adaptability to environmental changes and have identified conserved chromatin-associated factors that modulate levels of transcriptional noise. Herein, we show destabilization of pluripotency-associated gene regulatory networks through increased transcriptional heterogeneity of mouse embryonic stem cells in which paradigmatic histone acetyl-transferase, and candidate noise modulator, Kat2a (yeast orthologue Gcn5), have been inhibited. Functionally, network destabilization associates with reduced pluripotency and accelerated mesendodermal differentiation, with increased probability of transitions into lineage commitment. Thus, we show evidence of a relationship between transcriptional heterogeneity and cell fate transitions through manipulation of the histone acetylation landscape of mouse embryonic stem cells, suggesting a general principle that could be exploited in other normal and malignant stem cell fate transitions. Stem Cells 2018;36:1828-11.


Nitrite reductase is critical for Pseudomonas aeruginosa survival during co-infection with the oral commensal Streptococcus parasanguinis.
Pseudomonas aeruginosa is the major aetiological agent of chronic pulmonary infections in cystic fibrosis (CF) patients. However, recent evidence suggests that the polymicrobial community of the CF lung may also harbour oral streptococci, and colonization by these micro-organisms may have a negative impact on P. aeruginosa within the CF lung. Our previous studies demonstrated that nitrite abundance plays an important role in P. aeruginosa survival during co-infection with oral streptococci. Nitrite reductase is a key enzyme involved in nitrite metabolism. Therefore, the objective of this study was to examine the role nitrite reductase (gene nirS) plays in P. aeruginosa survival during co-infection with an oral streptococcus, Streptococcus parasanguinis. Inactivation of nirS in both the chronic CF isolate FRD1 and acute wound isolate PAO1 reduced the survival rate of P. aeruginosa when co-cultured with S. parasanguinis. Growth of both mutants was restored when co-cultured with S. parasanguinis that was defective for H2O2 production. Furthermore, the nitrite reductase mutant was unable to kill Drosophila melanogaster during co-infection with S. parasanguinis. Taken together, these results suggest that nitrite reductase plays an important role for survival of P. aeruginosa during co-infection with S. parasanguinis.


Effects of anticonvulsant therapy on recovery and rehabilitation.
Antiepileptic drugs (AEDs) are used in rehabilitation patients with injury to the nervous system for control of clinical seizures, prophylaxis against the development of a seizure disorder, and treatment of secondary mania. Although most AEDs can produce mild cognitive side effects, the benefits of treatment with AEDs in rehabilitation patients with seizure disorders typically outweigh these minor risks, Bromide, phenobarbital, and benzodiazepines may have more severe cognitive side effects than other AEDs. However, there is no convincing evidence of clinically significant differences in cognitive adverse effects among the other established AEDs. Furthermore, cognitive side effects are usually modest with mono therapy and anticonvulsant blood levels within standard therapeutic ranges. With regard to prophylaxis against future seizures, there is no significant data indicating that AED administration can prevent or retard the process of epileptogenesis, Therefore, the use of AEDs to prevent the development of a seizure disorder is not warranted at this time. Finally, although there is little information about the behavioral effects of anticonvulsants in neurological rehabilitation patients, the efficacy of carbamazepine, and perhaps valproate, in the treatment of primary mania suggests that these AEDs may be beneficial in mania secondary to brain injury. The role of AEDs in the treatment of other psychiatric disorders, such as post-traumatic stress disorder and episodic dyscontrol, remains unclear.


Tuning the Magnitude and the Polarity of the Piezoresistive Response of Polyaniline through Structural Control.
We demonstrate the tunability of both the polarity and the magnitude of the piezoresistive response of polyaniline that is template-synthesized on poly(2-acrylamido-2-methyl-1-propanesulfonic acid), PANI-PAAMPSA, by altering the template molecular weight. Piezoresistivity is quantified by gauge factor, a unitless parameter that relates changes in electrical resistance to applied strain. The gauge factor of PANI-PAAMPSA decreases linearly and becomes negative with decreasing PAAMPSA molecular weight. The polarity of PANI-PAAMPSA's gauge factor is determined by macroscopic connectivity across thin films. PANI-PAAMPSA thin films comprise electrostatically stabilized particles whose size is determined at the onset of synthesis. An increase in the interparticle spacing with applied strain results in a positive gauge factor. The presence of PANI crystallites increases connectivity between particles; these samples instead exhibit a negative gauge factor whereby the resistance decreases with increasing strain. The tunability of the piezoresistive response of these conducting polymers allows their utilization in a broad range of flexible electronics applications, including thermo- and chemoresistive sensors and strain gauges.


Identification and characterization of the PhhR regulon in Pseudomonas putida.
Pseudomonas putida is a soil microorganism that utilizes aromatic amino acids present in root exudates as a nitrogen source. We have previously shown that the PhhR transcriptional regulator induces phhAB genes encoding a phenylalanine hydroxylase. In this study we show, using microarray assays and promoter fusions, that PhhR is a global regulator responsible for the activation of genes essential for phenylalanine degradation, phenylalanine homeostasis and other genes of unknown function. Recently, it has been shown that phenylalanine catabolism occurs through more than one pathway. One of these possible pathways involves the metabolism of phenylalanine via tyrosine, p-hydroxyphenylpyruvate, and homogentisate. We identified two genes within this pathway that encode an acyl-CoA transferase involved in the metabolism of acetoacetate. All genes in this pathway were induced in response to phenylalanine in a PhhR-proficient background. The second potential degradative pathway involves the degradation of phenylalanine to produce phenylpyruvate, which seems to be degraded via phenylacetyl-CoA. A number of mutants in the paa genes encoding phenylacetyl-CoA degradation enzymes fail to grow on phenylpyruvate or phenylacetate, further supporting the existence of this second pathway. We found that the PhhR regulon also includes genes involved in the biosynthesis of aromatic amino acids that are repressed in the presence of phenylalanine, suggesting the possibility of feedback at the transcriptional level. In addition, we found that PhhR modulates the level of expression of the broad-substrate-specificity MexEF/OprN efflux pump. Expression from this pump is under the control of mexT gene product because phenylalanine-dependent transcription from the mexE promoter does not occur in a mexT mutant background. These results place PhhR as an important regulator in the control of bacterial responses to aromatic amino acids.


Pentacyclic triterpenes combined with ciprofloxacin help to eradicate the biofilm formed in vitro by Escherichia coli.
Ciprofloxacin is commonly used in clinical practice for the treatment of recurrent urinary tract infections caused by Escherichia coli. However, very often these recurrent infections are due to a failure in a complete eradication of the microorganisms colonizing the urinary tract, especially in catheterized patients. To enhance the bactericidal activity of ciprofloxacin against biofilm-forming uropathogenic E. coli (UPECs), we examined its effect in combination with two pentacyclic triterpenes - asiatic and ursolic acids. The anti-biofilm activity of ciprofloxacin and pentacyclic triterpenes - asiatic acid (AA) and ursolic acid (UA), as well as their synergistic effect were tested on two types of surfaces - polystyrene microtiter plates and silicone catheters. It was investigated using the time-killing and biofilm assays. a0 nti-biofilm activity of ciprofloxacin was not observed on microtiter plates or on the catheters. Ciprofloxacin combined with ursolic acid inhibited the biofilm formation on microtitre plates. This mixture, however, did not express such a strong activity against the synthesis of biofilm on the surface of catheters. Ciprofloxacin combined with asiatic acid had very weak inhibiting effect on the synthesis of biofilm mass on microtitre plates as well as on the catheters. Despite this, both mixtures - ciprofloxacin and asiatic acid, as well as ciprofloxacin and ursolic acid, exhibited strong and significant impact on the eradication of mature biofilm (P < 0.05). Although ciprofloxacin is recommended in the treatment of urinary tract infections caused by UPECs, but its efficacy is arguable. Subinhibitory concentrations of ciprofloxacin did not inhibit the formation of biofilm. Pentacyclic triterpenes used in combination with ciprofloxacin enhanced its anti-biofilm effectiveness. However, this anti-biofilm activity was found to depend on the type of surface on which biofilm was formed.


Bevacizumab-Based Chemotherapy Combined with Regional Deep Capacitive Hyperthermia in Metastatic Cancer Patients: A Pilot Study.
As an angiogenesis inhibitor, bevacizumab has been investigated in combination with different chemotherapeutic agents, achieving an established role for metastatic cancer treatment. However, potential synergic anti-angiogenic effects of hyperthermia have not tested to date in literature. The aim of our study was to analyze efficacy, safety, and survival of anti-angiogenic-based chemotherapy associated to regional deep capacitive hyperthermia (HT) in metastatic cancer patients. Twenty-three patients with metastatic colorectal (n = 16), ovarian (n = 5), and breast (n = 2) cancer were treated with HT in addition to a standard bevacizumab-based chemotherapy regimen. Treatment response assessment was performed, according to the modified Response Evaluation Criteria for Solid Tumors (mRECIST), at 80 days (timepoint-1) and at 160 days (timepoint-2) after therapy. Disease Response Rate (DRR), considered as the proportion of patients who had the best response rating (complete response (CR), partial response (PR), or stable disease (SD)), was assessed at timepoint-1 and timepoint-2. Chi-squared for linear trend test was performed to evaluated the association between response groups (R/NR) and the number of previous treatment (none, 1, 2, 3), number of chemotherapy cycles (<6, 6, 12, >12), number of hyperthermia sessions (<12, 12, 24, >24), and lines of chemotherapy (I, II). Survival curves were estimated by Kaplan-Meier method. DRR was 85.7% and 72.2% at timepoint-1 and timepoint-2, respectively. HT was well tolerated without additional adverse effects on chemotherapy-related toxicity. Chi-squared for linear trend test demonstrated that the percentage of responders grew in relation to the number of chemotherapy cycles (p = 0.015) and to number of HT sessions (p < 0.001) performed. Both overall survival (OS) and time to progression (TTP) were influenced by the number of chemotherapy cycles (p < 0.001) and HT sessions (p < 0.001) performed. Our preliminary data, that need to be confirmed in larger studies, suggest that the combined treatment of bevacizumab-based chemotherapy with HT has a favorable tumor response, is feasible and well tolerated, and offers a potentially promising option for metastatic cancer patients.


In Vitro Culture of the Insect Endosymbiont Spiroplasma poulsonii Highlights Bacterial Genes Involved in Host-Symbiont Interaction.
Endosymbiotic bacteria associated with eukaryotic hosts are omnipresent in nature, particularly in insects. Studying the bacterial side of host-symbiont interactions is, however, often limited by the unculturability and genetic intractability of the symbionts. Spiroplasma poulsonii is a maternally transmitted bacterial endosymbiont that is naturally associated with several Drosophila species. S. poulsonii strongly affects its host's physiology, for example by causing male killing or by protecting it against various parasites. Despite intense work on this model since the 1950s, attempts to cultivate endosymbiotic Spiroplasma in vitro have failed so far. Here, we developed a method to sustain the in vitro culture of S. poulsonii by optimizing a commercially accessible medium. We also provide a complete genome assembly, including the first sequence of a natural plasmid of an endosymbiotic Spiroplasma species. Last, by comparing the transcriptome of the in vitro culture to the transcriptome of bacteria extracted from the host, we identified genes putatively involved in host-symbiont interactions. This work provides new opportunities to study the physiology of endosymbiotic Spiroplasma and paves the way to dissect insect-endosymbiont interactions with two genetically tractable partners.IMPORTANCE The discovery of insect bacterial endosymbionts (maternally transmitted bacteria) has revolutionized the study of insects, suggesting novel strategies for their control. Most endosymbionts are strongly dependent on their host to survive, making them uncultivable in artificial systems and genetically intractable. Spiroplasma poulsonii is an endosymbiont of Drosophila that affects host metabolism, reproduction, and defense against parasites. By providing the first reliable culture medium that allows a long-lasting in vitro culture of Spiroplasma and by elucidating its complete genome, this work lays the foundation for the development of genetic engineering tools to dissect endosymbiosis with two partners amenable to molecular study. Furthermore, the optimization method that we describe can be used on other yet uncultivable symbionts, opening new technical opportunities in the field of host-microbes interactions.


Regulation of Chloroplast Protein Import by the Ubiquitin E3 Ligase SP1 Is Important for Stress Tolerance in Plants.
Chloroplasts are the organelles responsible for photosynthesis in plants [1, 2]. The chloroplast proteome comprises ∼3,000 different proteins, including components of the photosynthetic apparatus, which are highly abundant. Most chloroplast proteins are nucleus-encoded and imported following synthesis in the cytosol. Such import is mediated by multiprotein complexes in the envelope membranes that surround each organelle [3, 4]. The translocon at the outer envelope membrane of chloroplasts (TOC) mediates client protein recognition and early stages of import. The TOC apparatus is regulated by the ubiquitin-proteasome system (UPS) in a process controlled by the envelope-localized ubiquitin E3 ligase SUPPRESSOR OF PPI1 LOCUS1 (SP1) [5, 6]. Previous work showed that SP1-mediated regulation of chloroplast protein import contributes to the organellar proteome changes that occur during plant development (e.g., during de-etiolation). Here, we reveal a critical role for SP1 in plant responses to abiotic stress, which is a major and increasing cause of agricultural yield losses globally [7]. Arabidopsis plants lacking SP1 are hypersensitive to salt, osmotic, and oxidative stresses, whereas plants overexpressing SP1 are considerably more stress tolerant than wild-type. We present evidence that SP1 acts to deplete the TOC apparatus under stress conditions to limit the import of photosynthetic apparatus components, which may attenuate photosynthetic activity and reduce the potential for reactive oxygen species production and photo-oxidative damage. Our results indicate that chloroplast protein import is responsive to environmental cues, enabling dynamic regulation of the organellar proteome, and suggest new approaches for improving stress tolerance in crops.


The value of biochemical markers for risk stratification prior to hospital admission in acute chest pain.
We describe the use of biochemical markers in the pre-hospital setting with regard to diagnostic accuracy for the detection of an acute myocardial infarction (AMI) and for prognosis in connection with acute chest pain. The sensitivity has been reported to be limited; blood sampling occurs very early and often prior to the release of biochemical markers into the circulation. The specificity was in some studies also limited, but this is more difficult to explain. New biochemical markers like human heart fatty acid binding protein (H-FACB) have shown improved diagnostic accuracy, in the pre-hospital setting, in one small pilot study compared with traditional biochemical markers like troponins, creatine kinase (CK-MB) and myoglobin. However, in a recent small study, the sensitivity for troponin I (when a low decision limit for myocardial damage was used), when analysed prior to hospital admission, was reported to be very high. The latter data need to be confirmed in larger studies and various biochemical markers reflecting various pathophysiological aspects of the disease need to be tested before the analysis of any marker can be recommended for use in the pre-hospital setting of a suspected AMI.