Differential regulation of germline apoptosis in response to meiotic checkpoint activation.
In Caenorhabditis elegans, germline apoptosis is promoted by egl-1 and ced-13 in response to meiotic checkpoint activation. We report that the requirement for these two factors depends on which checkpoints are active. We also identify a regulatory region of egl-1 required to inhibit germline apoptosis in response to DNA damage incurred during meiotic recombination.


Noninvasive measurement of advanced glycation end-products in the facial skin: New data for skin aging studies.
Using skin autofluorescence (SAF) as a marker of advanced glycation end-products (AGEs) has been extensively studied in the last decade since the introduction of the noninvasive in vivo measurement technique. Data have shown the level of skin AGEs increases with chronological age in healthy human beings, and this increase is substantially higher in age-matched diabetic patients. In skin research, glycation with the accompanying accumulation of skin AGEs has been regarded as one of the primary skin aging mechanisms that contribute to skin wrinkling and the loss of skin elasticity. To date, the totality of SAF data reported in literature has been obtained from measurements on the arm, and noninvasive measurement of facial skin AGE accumulation would add great value to skin aging research. In this study, we report the levels of facial and forearm skin AGEs in 239 men and women of 21-65 year of age. Significantly lower levels of AGEs were detected in the facial skin than in the forearm skin from the young Caucasian groups, and the difference was much larger for men than for women. The rate of change in skin AGE level over age was found to be about 50% higher in men than in women, which further highlights the gender difference. A statistically significant correlation between the levels of skin AGE and facial wrinkling was also observed. The facial skin AGE data may provide new insight into skin aging research.


Structure and development of Medicago truncatula pod wall and seed coat.
Medicago truncatula has gained much attention as a genomic model species for legume biology, but little is known about the morphology of its pods and seeds. Structural and developmental characteristics of M. truncatula pod walls and seed coats are presented. Plants of M. truncatula ecotype A17 were grown under controlled conditions in a greenhouse. Flowers were date-tagged at anthesis, so that pods of known age could be collected. Harvested pods were fixed and sectioned for light microscopy. Structural attributes of pod walls and seed coats were characterized at four time points throughout early to mid-stages of pod development (3, 6, 13 and 20 d post-pollination). Basic features of the pod wall are an exocarp comprised of a single epidermal layer, a mesocarp with seven to 14 layers of parenchyma cells, and an endocarp composed of an inner epidermal cell layer and three to five layers of sclerenchyma cells adjacent to it. Vascular bundles are abundant in the pod wall and include one lateral carpellary bundle, one median carpellary bundle and nine to 12 vascular bundles, all embedded within the mesocarp parenchyma. Seed coat features include an epidermal layer of macrosclereids, a sub-epidermal layer of osteosclereids, and two to five rows of internal parenchyma cells. The hilar region contains the tracheid bar and the chalazal vascular bundle, the latter of which expands to form only two short branches. This characterization provides a needed understanding of pod structure and development in this model legume, and should facilitate various molecular investigations into legume fruit and seed biology.


Filter implementation technique for multicriteria characterization of coding domains in the joint transform correlator.
An improved method for implementing correlation filters in the joint transform correlator architecture is proposed. We derived the method from computer-generated holography techniques. It allows us to use any correlation filters, especially ones that provide an optimal trade-off between noise robustness, peak sharpness, and optical efficiency, with any spatial light modulator (SLM). This method also allows for an objective comparison of the performance of the coding domains of various SLM's.


The African Disability Scooter: preliminary analysis of a new mobility aid.
In many resource poor African countries there is no effective prosthetic service. The African Disability Scooter (ADS) was developed for use by amputees in such countries, as a positive alternative to crutches or sticks. The aim of this study was to compare the energy expenditure of the ADS to walking with crutches over a fixed distance at a self-selected speed in a normal population. Speed and efficiency of the scooter were assessed by field tests with 10 volunteers using a portable, energy consumption measuring device. In addition, the comfort of the scooter was assessed with visual analogue scales. It was found that the scooter was twice as efficient, and twice as fast as crutches over level ground. The scooter was also rated highly on comfort and usability scores, although some suggested refinements are likely to improve this further. The results suggest that the ADS is a potentially useful method of transport by people with amputations or a wide spectrum of unilateral lower limb disabilities in developed and developing countries.


Co-expression of myosin II regulatory light chain and the NMDAR1 subunit in neonatal and adult mouse brain.
Movement of glutamate receptors in neurons likely involves direct and indirect association of receptor subunits with microtubule- and actin-based motor proteins. We have previously shown that myosin II regulatory light chain (RLC) binds directly to subunits of the NMDA-type glutamate receptor (NR), suggesting that NMDA receptors are closely associated with a myosin II motor complex. Using a polyclonal antibody predicted to recognize all RLC isoforms previously described in rodent brain, we report the expression of RLC and the NR1 subunit in cortex, hippocampus and cerebellum of postnatal day 0 (P0) and adult mouse. Although myosin RLC was not exclusively localized with NR1 by immunohistochemistry, co-staining was striking in the neuronal soma of deep cortical neurons and Purkinje neurons of the cerebellum which showed a punctate, perinuclear pattern of immunoreactivity. These neuronal populations were identified using a monoclonal antibody directed against a nuclear-specific, transcriptional repressor, chicken ovalbumin upstream promoter-transcription factor (COUP-TF)-interacting protein 2 (CTIP2). Co-expression of NR1 and a myosin II motor was validated using an isoform specific anti-nonmuscle myosin II-B heavy chain (NMHC II-B) antibody. Our findings support the idea that there is regional heterogeneity in the molecular composition of the NMDA receptor-associated cytoskeleton, and suggest that NR subunits may be associated with an actin-based, myosin II-B motor within the endomembrane system of some neuronal populations. Differential staining patterns observed with light and heavy chain antibodies, however, suggest that there is also heterogeneity in the composition of myosin II complexes in brain.


Radiation-induced non-targeted effects: some open questions.
The existence of non-targeted effects (NTEs) of radiation (genomic instability and bystander effects) has been generally accepted for >20 y; however, there is research, which was largely ignored going back to 1915 reporting these effects. Despite today's general acceptance of the phenomenon of NTE, there is little agreement about the mechanisms involved and the implications in radiation biology and radiation protection. The aim of this review was to consider some of the odd data, which have been published in the field with a view to obtaining insights or stimulating new ways of thinking about this field. By highlighting some key challenges and controversies, concerning the mechanisms and more importantly, the reason these effects exist, current ideas about the wider implications of NTEs in evolution and biology are also discussed.


Sequential Therapy versus Triple Therapy for the First Line Treatment of Helicobacter pylori in Korea: A Nationwide Randomized Trial.
Eradication of Helicobacter pylori infection with standard triple therapy (TT) has declined primarily because of increased antibiotic resistance. Sequential therapy (ST) has been suggested as an alternative to TT for the first-line treatment of H. pylori. The purpose of this study was to compare the efficacy of ST with TT. This was a multicenter, randomized open-label trial performed at nine centers in Korea. Patients with H. pylori infection were randomly assigned to receive either 7 day TT or 10 day ST. Eradication rates, drug compliance, and adverse events were compared among the two regimens. A total of 601 patients were enrolled between March 2011 and September 2014. The intention-to-treat eradication rates were 70.8% for TT and 82.4% for ST (p=0.001). The corresponding per protocol eradication rates were 76.9% and 88.8% for TT and ST, respectively (p=0.000). There were no statistically significant differences between the two regimens with respect to drug compliance and adverse events. ST achieved better eradication rates than TT as a first-line therapy for H. pylori eradication in Korea.


Current Belgian adenotonsillectomy practice: a survey among Belgian ENT specialists.
Tonsillectomies and adenotonsillectomies (TE and ATE) are among the most frequently performed operations in the western world. Despite the lack of generally accepted clinical guidelines about indications, the beneficial role of the procedure has been established. The aim of this study was to identify current practice relating to indications for tonsillectomies and adenotonsillectomies among Belgian ENT specialists. An anonymous questionnaire was sent to all Belgian ENT specialists. Part I of the questionnaire included questions about the ENT specialist himself/herself and his/her practice. Part II comprised questions about indications for TE/ATE in children up to 16 years of age. Part III included questions about indications in adults. 238 out of 528 questionnaires were completed and returned. The majority of physicians consider TE/ATE to be indicated if a child or an adult suffers 3 or 4 tonsillitis episodes per year. The decision to operate on children for upper airway obstruction is mainly based on clinical and anamnestic findings. A peritonsillar abscess is usually considered to be an indication for surgery. We compared the results between different groups of ENT specialists and could detect differences in decision-making only according to the number of years of ENT practice. The stringency of surgeons' indications decreases with increasing surgeon age. The majority of respondents consider tonsillectomies/adenotonsillectomies to be indicated after 3 to 4 episodes of tonsillitis per year. In the case of upper airway obstruction in children, surgical indications are mainly based on clinical and anamnestic findings. Surgical decision-making appears to be influenced by the number of years of ENT practice.


An observational study fluid balance and patient outcomes in the Randomized Evaluation of Normal vs. Augmented Level of Replacement Therapy trial.
To examine associations between mean daily fluid balance during intensive care unit study enrollment and clinical outcomes in patients enrolled in the Randomized Evaluation of Normal vs. Augmented Level (RENAL) replacement therapy study. Statistical analysis of data from multicenter, randomized, controlled trials. Thirty-five intensive care units in Australia and New Zealand. Cohort of 1453 patients enrolled in the RENAL study. We analyzed the association between daily fluid balance on clinical outcomes using multivariable logistic regression, Cox proportional hazards, time-dependent analysis, and repeated measure analysis models. During intensive care unit stay, mean daily fluid balance among survivors was -234 mL/day compared with +560 mL/day among nonsurvivors (p < .0001). Mean cumulative fluid balance over the same period was -1941 vs. +1755 mL (p = .0003). A negative mean daily fluid balance during study treatment was independently associated with a decreased risk of death at 90 days (odds ratio 0.318; 95% confidence interval 0.24-0.43; p < .000.1) and with increased survival time (p < .0001). In addition, a negative mean daily fluid balance was associated with significantly increased renal replacement-free days (p = .0017), intensive care unit-free days (p < .0001), and hospital-free days (p = .01). These findings were unaltered after the application of different statistical models. In the RENAL study, a negative mean daily fluid balance was consistently associated with improved clinical outcomes. Fluid balance may be a target for specific manipulation in future interventional trials of critically ill patients receiving renal replacement therapy.


Streptozotocin-induced β-cell damage, high fat diet, and metformin administration regulate Hes3 expression in the adult mouse brain.
Diabetes mellitus is a group of disorders characterized by prolonged high levels of circulating blood glucose. Type 1 diabetes is caused by decreased insulin production in the pancreas whereas type 2 diabetes may develop due to obesity and lack of exercise; it begins with insulin resistance whereby cells fail to respond properly to insulin and it may also progress to decreased insulin levels. The brain is an important target for insulin, and there is great interest in understanding how diabetes affects the brain. In addition to the direct effects of insulin on the brain, diabetes may also impact the brain through modulation of the inflammatory system. Here we investigate how perturbation of circulating insulin levels affects the expression of Hes3, a transcription factor expressed in neural stem and progenitor cells that is involved in tissue regeneration. Our data show that streptozotocin-induced β-cell damage, high fat diet, as well as metformin, a common type 2 diabetes medication, regulate Hes3 levels in the brain. This work suggests that Hes3 is a valuable biomarker helping to monitor the state of endogenous neural stem and progenitor cells in the context of diabetes mellitus.


Effect of ocean acidification on the nutritional quality of marine phytoplankton for copepod reproduction.
Phytoplankton are the oceans' principal source of polyunsaturated fatty acids that support the growth and reproduction of consumers such as copepods. Previous studies have demonstrated ocean acidification (OA) can change the availability of polyunsaturated fatty acids to consumer diets which may affect consumer reproduction. Two laboratory experiments were conducted to examine the effects of feeding high-pCO2-reared phytoplankton on copepod egg production, hatching success, and naupliar survival. Marine phytoplankton Rhodomonas salina, Skeletonema marinoi, Prorocentrum micans, and Isochrysis galbana were exponentially grown in semi-continuous cultures at present (control) (400 ppm CO2, pH~8.1) and future (1,000 ppm CO2, pH~7.8) conditions and provided to Acartia tonsa copepods over 4 consecutive days as either nitrogen-limited (Exp. I) or nitrogen-depleted (Exp. II) mixed assemblage of phytoplankton. The composition of FAs in the phytoplankton diet was affected by pCO2 concentration and nitrogen deficiency; the ratio of essential fatty acids to total polyunsaturated fatty acids decreased in phytoplankton grown under high pCO2 and the mass of total fatty acids increased under nitrogen depletion. Additionally, total concentrations of essential fatty acids and polyunsaturated fatty acids in the diet mixtures were less under the high-pCO2 compared to the control-pCO2 treatments. Median egg production, hatching success, and naupliar survival were 48-52%, 4-87%, and 9-100% lower, respectively, in females fed high-pCO2 than females fed low-pCO2 phytoplankton, but this decrease in reproductive success was less severe when fed N-depleted, but fatty acid-rich cells. This study demonstrates that the effects of OA on the nutritional quality of phytoplankton (i.e., their cellular fatty acid composition and quota) were modified by the level of nitrogen deficiency and the resulting negative reproductive response of marine primary consumers.


Validation of a simple histological-histochemical cartilage scoring system.
In this study, we assessed the validity of a subjective histological-histochemical scoring system as compared to an automated histomorphometry program for analyzing cartilage repair tissue. In the first part of the study, we assessed the ability of the human eye to estimate the percent cartilage in a histological section. Twenty-nine rabbit periosteal explants that had been cultured in agarose transforming growth factor-beta (TGF-beta) were selected so that the percentage of cartilage in the specimens was distributed equally from 0% to 100%. Color photomicrographs were evaluated by 5 expert observers who gave a visual estimate of the percent cartilage. There was a strong correlation between the estimated and actual percent cartilage (R(2) = 0.92, p < 0.0001) and among the observers (I.C.C. = 0.89). On average, the estimated percent cartilage was within ten percent of the actual percent measured. In the second part, we compared the data derived using a simple cartilage score with those obtained by automated image analysis. The histological slides from 159 explants cultured under various experimental conditions (14 treatment groups) in two different experiments were analyzed. The cartilage content was estimated visually and a score from 0 to 3 was assigned. A previously validated, computerized image analysis system was used to measure the actual percent cartilage. Statistical analyses revealed a good linear regression (R(2) = 0.84, p = 0.0001), and even better polynomial correlation between the actual measurement and the score (R(2) = 0.88, p = 0.0001). These data demonstrate the validity of a simple histological-histochemical subjective scoring system. A computerized automated program such as the one employed in this study is preferable due to its many advantages. However, a subjective scoring system may be appropriate to use when the funding and expertise required for a computerized image analysis program are not available.


Characterization of Post-Thrombotic Syndrome in Children with Cardiac Disease.
To assess the validity of existing clinical scales assessing the presence of physical and functional abnormalities for diagnosing post-thrombotic syndrome (PTS) in children, including specific evaluation of use in children with congenital heart disease (CHD). One hundred children aged >2 years (average age, 6 years), including 33 with CHD and previously proven extremity deep vein thrombosis (DVT), 37 with CHD and no previous DVT, and 30 healthy siblings, were blindly assessed for PTS using the modified Villalta Scale (MVS). All patients aged <6 years underwent neurodevelopmental testing and an age-appropriate quality of life assessment. The MVS identified mild PTS in 20 children and moderate PTS in 1 child (including 14 of 33 [42%] in the CHD/DVT group, 5 of 37 [14%] in the CHD/no DVT group, and 2 of 30 controls [7%]). The diagnosis of PTS was confirmed clinically in 14 patients, all of whom had previous thrombosis and 1 of whom was MVS-negative. MVS had an accuracy of 91% and performed reasonably well as a screening tool but poorly as a diagnostic tool. MVS reliability was acceptable. Children with PTS had similar quality of life as those without PTS but had higher rates of neurodevelopmental delays in gross motor skills (70% vs 24%; P = .02) and problem-solving indicators (60% vs 15%; P = .008). Using the MVS scale for PTS screening in children with CHD is feasible and reliable, and the scale has good correlation with a clinical diagnosis of PTS despite a high prevalence of false-positive findings. Further research is needed to determine the clinical relevance of PTS in this population.


Parkinson's disease as a neuroendocrine disorder of circadian function: dopamine-melatonin imbalance and the visual system in the genesis and progression of the degenerative process.
For more than 50 years, Parkinson's disease (PD) has been conceptualized as a product of nigro-striatal dopamine (NSD) system degeneration. In spite of a growing body of evidence depicting the mammalian brain as an interrelated complexity of circuitous systems, dopamine (DA) deficiency of the NSD is still regarded as the main problem, with DA replacement being the purpose of therapeutic intervention. For at least 191 years circadian involvement in various aspects of PD, including depression and insomnia, has been recognized as an integral part of the symptom matrix of PD and yet attempts to elucidate the involvement of this system is uncharted territory. The present review attempts a major reorganization of mammalian brain into a coordinated complex involving the NSD and the retinal hypothalamic tract (RHT) as the primary systems involved in the retino-diencephalic/mesencephalic-pineal (RDMP) axis. Secondary systems including the lateral hypothalamus (LH), the area postraema (AP) and the subthalamic nucleus (STN) also form an integral part of this system as they have been shown to be either intimately related to the primary systems of the RDMP axis or have been shown to be significantly involved in the expression and treatment of PD. A large volume of evidence suggests that the RDMP axis is activated during the course of PD and during therapeutic intervention. Four types of neurotoxicity associated with melatonin are identified and the susceptibility of various parts of the RDMP axis to undergo neuropathological change, the tendency for melatonin to induce PD-like behavioural toxicity, and the relationship of this to PD symptomotology are described. This includes adverse effects of melatonin on motor function, hypotension, the adjuvant use of benzodiazepines, depression, insomnia, body weight regulation and various biochemical effects of melatonin administration: all problems currently facing the proposal to introduce melatonin as an adjuvant. It is suggested further that traditional DA replacement may well work by exerting its effect upon the circadian system, rather than simply replacing deficient DA. Activation of the circadian function by antagonizing melatonin with bright light not only has therapeutic value in treating the primary symptoms of PD but it shares a common mechanism with L-dopa in reducing the occurrence of seborrheic dermatitis. Concepts at the centre of understanding pineal function in PD, including pineal calcification, melatonin deficiency, symptomatic versus protective features of melatonin and antioxidative effects, are explained in a counterintuitive context. Intriguing propositions including the role of the retina in the aetiology of PD and that the nigra functions as a retina in this disorder are presented with the intention to provide a new understanding of the underlying compromised function in PD and to provide new treatment strategies. For the first time, abundant evidence is presented describing PD as an endocrine disorder of melatonin hyperplasia. The role of circadian interventive therapies and internal desynchrony in the aetiology and progression of PD provides a new direction for understanding the underlying physiology of a disease which is currently in a state of impasse and provides new hope for those who suffer from its debilitating effects.


Use of the Simple and Extended Grunwald-Winstein Equations in the Correlation of the Rates of Solvolysis of Highly Hindered Tertiary Alkyl Derivatives.
The original Grunwald-Winstein equation (1948) involved the development of a scale of solvent ionizing power (Y). Subsequent work has refined this scale and involved the development of scales of solvent nucleophilicity (N) and a term to correct for deviations when aromatic rings are present, governed by the aromatic ring parameter (I). These three scales, and the sensitivities towards each, can be related to specific rates of solvolysis through linear free energy relationships (LFERs).One important area of application of LFERs has been to the solvolyses of tert-alkyl halides. It has been proposed that the solvolysis of tert-butyl chloride involves a nucleophilic component, although other workers have suggested that the effects observed are related to electrophilic not nucleophilic influences. Takeuchi (1997) studied a compound with two of the methyl groups of tert-butyl chloride replaced by neopentyl groups. For this highly-hindered substrate there was no evidence for nucleophilic participation. Liu (1998) and Takeuchi (2001) have reported concerning the solvolyses of additional significantly-hindered tertiary alkyl chlorides. Liu (2009) has presented a parallel study of bromides. Martins (2008) has considered hindered tertiary alkyl halides, mainly with carbon-carbon multiple bonds as substituents. It was proposed that the hI term was of importance, with the sensitivities (h) sometimes positive and sometimes negative. To explain negative values, it was suggested that the I scale might contain a nucleophilicity component. In this review, we bring together, with analysis and commentary, the work of Takeuchi, Liu, Martins and others concerning the solvolyses of tertiary alkyl halides, with emphasis on the relevance of the three scales that have been developed for use in Grunwald-Winstein correlations.


Effect of atorvastatin on testosterone levels.
Statins are one of the most prescribed classes of drugs worldwide. Atorvastatin, the most prescribed statin, is currently used to treat conditions such as hypercholesterolaemia and dyslipidaemia. By reducing the level of cholesterol, which is the precursor of the steroidogenesis pathway, atorvastatin may cause a reduction in levels of testosterone and other androgens. Testosterone and other androgens play important roles in biological functions. A potential reduction in androgen levels, caused by atorvastatin might cause negative effects in most settings. In contrast, in the setting of polycystic ovary syndrome (PCOS), reducing excessive levels of androgens with atorvastatin could be beneficial. Primary objective To quantify the magnitude of the effect of atorvastatin on total testosterone in both males and females, compared to placebo or no treatment. Secondary objectives To quantify the magnitude of the effects of atorvastatin on free testosterone, sex hormone binding globin (SHBG), androstenedione, dehydroepiandrosterone sulphate (DHEAS) concentrations, free androgen index (FAI), and withdrawal due to adverse effects (WDAEs) in both males and females, compared to placebo or no treatment. The Cochrane Hypertension Information Specialist searched the following databases for randomized controlled trials (RCTs) up to 9 November 2020: the Cochrane Hypertension Specialised Register; the Cochrane Central Register of Controlled Trials (CENTRAL); MEDLINE; Embase; ;two international trials registries, and the websites of the US Food and Drug Administration, the European Patent Office and the Pfizer pharmaceutical corporation. These searches had no language restrictions. We also contacted authors of relevant articles regarding further published and unpublished work. RCTs of daily atorvastatin for at least three weeks, compared with placebo or no treatment, and assessing change in testosterone levels in males or females. Two review authors independently screened the citations, extracted the data and assessed the risk of bias of the included studies. We used the mean difference (MD) with associated 95% confidence intervals (CI) to report the effect size of continuous outcomes,and the risk ratio (RR) to report effect sizes of the sole dichotomous outcome (WDAEs). We used a fixed-effect meta-analytic model to combine effect estimates across studies, and risk ratio to report effect size of the dichotomous outcomes. We used GRADE to assess the certainty of the evidence. We included six RCTs involving 265 participants who completed the study and their data was reported. Participants in two of the studies were male with normal lipid profile or mild dyslipidaemia (N = 140); the mean age of participants was 68 years. Participants in four of the studies were female with PCOS (N = 125); the mean age of participants was 32 years. We found no significant difference in testosterone levels in males between atorvastatin and placebo, MD -0.20 nmol/L (95% CI -0.77 to 0.37). In females, atorvastatin may reduce total testosterone by -0.27 nmol/L (95% CI -0.50 to -0.04), FAI by -2.59 nmol/L (95% CI -3.62 to -1.57), androstenedione by -1.37 nmol/L (95% CI -2.26 to -0.49), and DHEAS by -0.63 μmol/l (95% CI -1.12 to -0.15). Furthermore, compared to placebo, atorvastatin increased SHBG concentrations in females by 3.11 nmol/L (95% CI 0.23 to 5.99). We identified no studies in healthy females (i.e. females with normal testosterone levels) or children (under age 18). Importantly, no study reported on free testosterone levels. We found no significant difference between atorvastatin and placebo on the levels of total testosterone in males. In females with PCOS, atorvastatin lowered the total testosterone, FAI, androstenedione, and DHEAS. The certainty of evidence ranged from low to very low for both comparisons. More RCTs studying the effect of atorvastatin on testosterone are needed.


Current concepts and future directions in the diagnosis and management of lymphatic vascular disease.
Despite the central, complex role for the lymphatic system in the maintenance of human health, the biology of this important and complex vasculature has been relatively under-investigated. However, the last decade has witnessed a substantial growth in the elucidation of lymphatic structural biology and the function of this system in health and in disease. These newly gained insights can be used to formulate our evolving concepts about the diagnostic and therapeutic approaches to patients with lymphatic vascular disorders. In lymphedema, there is a spectrum of disease that extends from primary (heritable) to secondary (acquired) causes. Once detected, the presence of lymphatic edema mandates very specific modalities of intervention, predominated by physiotherapeutic techniques. In addition, a physiological basis for adjunctive, intermittent pneumatic compression has been established, and these modalities may be indicated in selected patient populations. The acknowledgement of a unique biology in lymphatic edemas is, increasingly, guiding research efforts within this field. Increasing investigative attention is being directed toward animal models of lymphatic vascular disease. As insight into the complex biology of the lymphatic vasculature continues to expand through focused biomedical investigation, the translation of these mechanistic insights into targeted, rationally conceived therapeutics will become increasingly feasible.


Health sector spending and spending on HIV/AIDS, tuberculosis, and malaria, and development assistance for health: progress towards Sustainable Development Goal 3.
Sustainable Development Goal (SDG) 3 aims to "ensure healthy lives and promote well-being for all at all ages". While a substantial effort has been made to quantify progress towards SDG3, less research has focused on tracking spending towards this goal. We used spending estimates to measure progress in financing the priority areas of SDG3, examine the association between outcomes and financing, and identify where resource gains are most needed to achieve the SDG3 indicators for which data are available. We estimated domestic health spending, disaggregated by source (government, out-of-pocket, and prepaid private) from 1995 to 2017 for 195 countries and territories. For disease-specific health spending, we estimated spending for HIV/AIDS and tuberculosis for 135 low-income and middle-income countries, and malaria in 106 malaria-endemic countries, from 2000 to 2017. We also estimated development assistance for health (DAH) from 1990 to 2019, by source, disbursing development agency, recipient, and health focus area, including DAH for pandemic preparedness. Finally, we estimated future health spending for 195 countries and territories from 2018 until 2030. We report all spending estimates in inflation-adjusted 2019 US$, unless otherwise stated. Since the development and implementation of the SDGs in 2015, global health spending has increased, reaching $7·9 trillion (95% uncertainty interval 7·8-8·0) in 2017 and is expected to increase to $11·0 trillion (10·7-11·2) by 2030. In 2017, in low-income and middle-income countries spending on HIV/AIDS was $20·2 billion (17·0-25·0) and on tuberculosis it was $10·9 billion (10·3-11·8), and in malaria-endemic countries spending on malaria was $5·1 billion (4·9-5·4). Development assistance for health was $40·6 billion in 2019 and HIV/AIDS has been the health focus area to receive the highest contribution since 2004. In 2019, $374 million of DAH was provided for pandemic preparedness, less than 1% of DAH. Although spending has increased across HIV/AIDS, tuberculosis, and malaria since 2015, spending has not increased in all countries, and outcomes in terms of prevalence, incidence, and per-capita spending have been mixed. The proportion of health spending from pooled sources is expected to increase from 81·6% (81·6-81·7) in 2015 to 83·1% (82·8-83·3) in 2030. Health spending on SDG3 priority areas has increased, but not in all countries, and progress towards meeting the SDG3 targets has been mixed and has varied by country and by target. The evidence on the scale-up of spending and improvements in health outcomes suggest a nuanced relationship, such that increases in spending do not always results in improvements in outcomes. Although countries will probably need more resources to achieve SDG3, other constraints in the broader health system such as inefficient allocation of resources across interventions and populations, weak governance systems, human resource shortages, and drug shortages, will also need to be addressed. The Bill & Melinda Gates Foundation.


Women in resource-poor settings are at risk of inadequate intakes of multiple micronutrients.
A systematic review was conducted to identify all studies that were published between 1988 and 2008 reporting micronutrient intakes of women in resource-poor settings. Inclusion criteria were study location (resource-poor), dietary assessment method (24-h recall, estimated/weighed record, or locally validated FFQ), energy and 1 or more micronutrient intakes reported (vitamin A, vitamin B-6, vitamin B-12, vitamin C, thiamin, riboflavin, niacin, folate, iron, or zinc), age range (15-50 y), sample size (≥30), and sex (female). Of the 1560 papers identified, 52 papers were included. Results showed that, except for vitamin A (29%), vitamin C (34%), and niacin (34%), the reported mean/median intakes in over 50% of studies were below the Estimated Average Requirement (EAR). Folate intake was most often below EAR (91% of studies). Regional differences were apparent for intakes of vitamins A, C, and B-6 and riboflavin; mean/median intakes in Latin America exceeded the EAR, whereas in Asia, reported mean/median intakes of vitamin C, vitamin A, and riboflavin were below the EAR in 47, 50, and 77% of the studies, respectively, as was the case for vitamin B-6 in 75% of the studies in Africa. These results suggest that inadequate intakes of multiple micronutrients are common among women living in resource-poor settings and emphasize the need for increased attention to the quality of women's diets. There is a need for more high-quality studies of women's micronutrient intakes.


The Fifth Carlos E. Rubio Memorial Lecture. Sclerosing cholangitis: pathogenesis, pathology, and practice.
Primary sclerosing cholangitis is a generally progressive, sometimes fatal, chronic hepatobiliary disorder for which no effective medical or surgical therapy now exists. The syndrome occurs most frequently in young men and is characterized by chronic cholestasis, frequent association with CUC, a paucity of serologic markers, hepatic copper overload, and characteristic abnormalities in some liver biopsy specimens and in virtually all cholangiograms. The natural history of the syndrome is still somewhat unclear; the disease likely progresses slowly and relentlessly over a decade or longer from an asymptomatic stage to a condition characterized by symptoms of cholestasis and complicated by cirrhosis and portal hypertension and carcinoma of the bile ducts. Management should first involve a thoughtful decision to observe, which is reasonable in the asymptomatic patient with early disease, or to intervene, particularly in patients with symptoms. Therapeutic goals should be defined and should concentrate on either alleviating symptoms, dealing with complications, or attempting to affect the underlying hepatobiliary disease. Symptomatic treatment and therapy for complications is similar to that employed in other chronic liver diseases, but also involves balloon dilatation of dominant strictures in appropriately selected symptomatic patients. Biliary tract reconstructive surgery may alleviate symptoms in selected patients with PSC, but its effect on the natural history of the syndrome has not been determined. Proctocolectomy for CUC in a patient with CUC and PSC does not beneficially affect the progression of the underlying hepatobiliary disease. In contrast, orthotopic liver transplantation may be life-saving for patients with advanced disease. Medical therapy directed at arresting the progression of the underlying hepatobiliary disease is currently experimental and includes cupruretic, immunosuppressive, antifibrogenic, and choleretic agents. Although a single recently completed controlled trial makes it unlikely that cupruretic agents will be helpful in this syndrome, immunosuppressive (i.e. cyclosporin A and methotrexate) and choleretic (i.e. ursodeoxycholic acid) agents alone or in combination are currently undergoing evaluation in randomized trials.


Factors and barriers influencing influenza vaccination among students at Brigham Young University.
To identify the prevalence of influenza vaccination and factors associated with vaccination among students at Brigham Young University. A Cross-sectional survey of seven general education classes, size 30 to 200 students each, was conducted the week of November 25, 2007. A 34 item paper-pencil questionnaire was administered, taking 5-10 minutes to complete. The response rate was 90%, with 421 completed surveys. Prevalence of influenza vaccination was 12% in the current influenza season. Influenza vaccination was significantly influenced by place of work, frequency of being around children, place of residence, and selected area of academic study. Students that received the influenza vaccination were more motivated by perceived severity of influenza than by perceived risk of contracting the illness. Physicians or nurses were the most influential at encouraging influenza vaccination, followed by parents, then the university or student health center, and then the media. The percentage of students that received influenza vaccination information from physicians or nurses was 14%, from parents was 15%, from the student health center was 25%, and from the general media was 45%. Influenza vaccination is low among college students, but impacted by perceived severity of the illness, place of employment or residence, and who encourages influenza vaccination.


Recommendations for Development of Botanical Polyphenols as "Natural Drugs" for Promotion of Resilience Against Stress-Induced Depression and Cognitive Impairment.
Extensive evidence has demonstrated that psychological stress has detrimental effects on psychological health, cognitive function, and ultimately well-being. While stressful events are a significant cause of psychopathology, most individuals exposed to adversity maintain normal psychological functioning. The mechanisms underlying such resilience are poorly understood, and there is an urgent need to identify and target these mechanisms to promote resilience under stressful events. Botanicals have been used throughout history to treat various medical conditions; however, the development of botanical compounds into potential preventative and therapeutic agents in studies promoting brain health is hindered by the fact that most orally consumed botanicals are extensively metabolized during absorption and/or by post-absorptive xenobiotic metabolism. Therefore, the primary objective of this review article is to provide recommendations for developing natural compounds as novel therapeutic strategies to promote resilience in susceptible subjects. The development of botanical polyphenols to ultimately attenuate mood disorders and cognitive impairment will rely on understanding (1) the absorption and bioavailability of botanical polyphenols with emphasis on flavan-3-ols, (2) the characterization of tissue-specific accumulation of biologically available polyphenols and their mechanisms of action in the brain, and eventually (3) the characterization of biologically available polyphenol metabolites in mechanisms associated with the promotion of resilience against mood disorders and cognitive impairment in response to stress. We also summarize exciting new lines of investigation about the role of botanicals such as polyphenols in the promotion of cognitive and psychological resilience. This information will provide a strategical framework for the future development of botanicals as therapeutic agents to promote resilience, ultimately preventing and/or therapeutically treating cognitive impairment and psychological dysfunction.


Sorption and biodegradation of vapor-phase organic compounds with wastewater sludge and food waste compost.
To test the possible use of composted food waste and wastewater sludge as biofilters to treat gas-phase volatile organic compounds (VOCs), batch experiments were conducted with an isolated strain that could degrade aromatic compounds under aerobic conditions. A benzene and trichloroethylene (TCE) mixture was used as the gas-phase pollutant in experiments with composted food waste, sludge, and soil. Under aerobic conditions, benzene was degraded as a primary substrate and TCE was degraded cometabolically, with water contents varying from 6 to 60% (volume of water added/volume of solid). Optimal water content for VOC removal was 12% for the soil, 36% for the composted food waste, and 48% for the sludge. The extent of VOC sorption and biodegradation at the optimal water content was different for each material. With the same initial VOC concentration, more VOCs were removed by sorption onto the composted food waste and the sludge, while less VOCs were biodegraded in comparison with the results using soil. The reason the biodegradation in the soil was greater may be partly attributed to the fact that, due to less sorption, the aqueous-phase concentration of VOCs, which microorganisms could utilize as a carbon source or cometabolize, was higher. We also speculate that the distribution of microorganisms in each medium affects the rate of biodegradation. A large number of microorganisms were attached to the composted food waste and sludge. Mass transfer of VOCs and oxygen to these microorganisms, which appear to have been heterogeneously distributed in clusters, may have been limited, resulting in hindered biodegradation.


Happiness: before and after the kids.
Understanding how having children influences parents' subjective well-being ("happiness") has great potential to explain fertility behavior. We study parental happiness trajectories before and after the birth of a child, using large British and German longitudinal data sets. We account for unobserved parental characteristics using fixed-effects models and study how sociodemographic factors modify the parental happiness trajectories. Consistent with existing work, we find that happiness increases in the years around the birth of a first child and then decreases to before-child levels. Moreover, happiness increases before birth, suggesting that the trajectories may capture not only the effect of the birth but also the broader process of childbearing, which may include partnership formation and quality. Sociodemographic factors strongly modify this pattern. Those who have children at older ages or who have more education have a particularly positive happiness response to a first birth; and although having the first two children increases happiness, having a third child does not. The results, which are similar in Britain and Germany, suggest that having up to two children increases happiness, and mostly for those who have postponed childbearing. This pattern is consistent with the fertility behavior that emerged during the second demographic transition and provides new insights into low and late fertility.


Theoretical model for double pulse laser-induced breakdown spectroscopy.
We present a simple theoretical model for the emission from double pulse laser-induced plasmas that was developed to better understand the processes and factors involved in enhancement of plasma emission. In this model, the plasma emission is directly proportional to the square of plasma density, its volume, and the fraction of second laser pulse absorbed through inverse bremsstrahlung absorption by the plasma plume of the first laser pulse. The electron-ion collision frequency determines the profile and location of the peak of emission enhancement with respect to the delay between the two lasers, whereas the amplitude of the enhancement is mainly dependent on the increase in the mass ablation rate after the second laser pulse. The effects of increase in temperature and in plasma volume after the second laser pulse are also discussed in light of this model.


Improving artificial metalloenzymes' activity by optimizing electron transfer.
While many artificial metalloenzymes have been reported, and are proposed to be highly promising for energy, environmental and medical applications, few could match the turnover rate (TOR) and turnover number (TON) of natural enzymes. Since electron transfer is oftentimes the rate-determining step, optimizing the electron transfer efficiency is an effective approach to significantly enhance artificial enzymes' activity. In this article, we review the recent progress in improving artificial metalloenzymes' activity by optimizing electron transfer.


Attachment to parents, social anxiety, and close relationships of female students over the transition to college.
The current study examined the process by which attachment to parents influences satisfaction with and ease in forming friendships at college. One hundred seventy-two female college freshmen completed a measure of parental attachment security the summer before their first semester of college (July 2006) and measures to assess satisfaction with and ease in forming close relationships at the end of their first semester (December 2006). Students ranged in age from 18 to 20 years (M = 18.09, SD = 0.33) and were diverse in their racial makeup (30% racial minority). Consistent with predictions derived from attachment theory, secure attachment to parents was positively associated with ease in forming friendships among racial minority and white participants and satisfaction with friendships among minority participants. Moreover, indirect effects of parental attachment security on relationship outcomes through social anxiety were significant for minority participants but not for white participants. Findings may be useful in the development of retention programs targeted at incoming university freshmen, particularly minority students.


Direct evidence for microbial-derived soil organic matter formation and its ecophysiological controls.
Soil organic matter (SOM) and the carbon and nutrients therein drive fundamental submicron- to global-scale biogeochemical processes and influence carbon-climate feedbacks. Consensus is emerging that microbial materials are an important constituent of stable SOM, and new conceptual and quantitative SOM models are rapidly incorporating this view. However, direct evidence demonstrating that microbial residues account for the chemistry, stability and abundance of SOM is still lacking. Further, emerging models emphasize the stabilization of microbial-derived SOM by abiotic mechanisms, while the effects of microbial physiology on microbial residue production remain unclear. Here we provide the first direct evidence that soil microbes produce chemically diverse, stable SOM. We show that SOM accumulation is driven by distinct microbial communities more so than clay mineralogy, where microbial-derived SOM accumulation is greatest in soils with higher fungal abundances and more efficient microbial biomass production.


Proteomic Analysis of Urine from California Sea Lions ( Zalophus californianus): A Resource for Urinary Biomarker Discovery.
Urinary markers for the assessment of kidney diseases in wild animals are limited, in part, due to the lack of urinary proteome data, especially for marine mammals. One of the most prevalent kidney diseases in marine mammals is caused by Leptospira interrogans, which is the second most common etiology linked to stranding of California sea lions ( Zalophus californianus). Urine proteins from 11 sea lions with leptospirosis kidney disease and eight sea lions without leptospirosis or kidney disease were analyzed using shotgun proteomics. In total, 2694 protein groups were identified, and 316 were differentially abundant between groups. Major urine proteins in sea lions were similar to major urine proteins in dogs and humans except for the preponderance of resistin, lysozyme C, and PDZ domain containing 1, which appear to be over-represented. Previously reported urine protein markers of kidney injury in humans and animals were also identified. Notably, neutrophil gelatinase-associated lipocalin, osteopontin, and epidermal fatty acid binding protein were elevated over 20-fold in the leptospirosis-infected sea lions. Consistent with leptospirosis infection in rodents, urinary proteins associated with the renin-angiotensin system were depressed, including neprilysin. This study represents a foundation from which to explore the clinical use of urinary protein markers in California sea lions.


Motion effects on intrusion development.
Analogue studies on intrusion development have found that visuospatial tasks performed during the encoding of aversive information reduce subsequent intrusion development. However, these studies cannot rule out a physical explanation in terms of simple movement. In the current study we addressed this issue. Healthy participants viewed an aversive film while performing a visuospatial movement task, a configurational movement task, or no task. Intrusive images from the film were reported in a diary during the week following film viewing. In line with an information-processing account of posttraumatic stress disorder, intrusion frequency was significantly reduced by the visuospatial movement task but not the configurational movement task compared to no task. This finding supports the role of visuospatial processing specifically in intrusion development.


Stability properties of periodically driven overdamped pendula and their implications to physics of semiconductor superlattices and Josephson junctions.
We consider the first order differential equation with a sinusoidal nonlinearity and periodic time dependence, that is, the periodically driven overdamped pendulum. The problem is studied in the case that the explicit time dependence has symmetries common to pure ac-driven systems. The only bifurcation that exists in the system is a degenerate pitchfork bifurcation, which describes an exchange of stability between two symmetric nonlinear modes. Using a type of Prüfer transform to a pair of linear differential equations, we derive an approximate condition of the bifurcation. This approximation is in very good agreement with our numerical data. In particular, it works well in the limit of large drive amplitudes and low external frequencies. We demonstrate the usefulness of the theory applying it to the models of pure ac-driven semiconductor superlattices and Josephson junctions. We show how the knowledge of bifurcations in the overdamped pendulum model can be utilized to describe the effects of rectification and amplification of electric fields in these microstructures.


Relation between stress and symptoms of craniomandibular disorders in adolescents.
The psychophysiologic theory proposes that stress can precipitate craniomandibular disorders (CMD) and that stress correlates more strongly to disorders of the masticatory muscles than to temporomandibular joint disorders. Empirical reports show only low correlations between emotional stress and CMD signs and symptoms, and that some of them might be spurious. In the present study this correlation was assessed in 417 adolescents from 11 to 16 years old. Data from the clinical examination were used to construct two indices: 1) The number of muscles sites tender to palpation, and 2) signs from the joint and restricted movement. Results show that global stress was only significantly correlated with the muscle index (r = .20), but not with the other index. Only the multiple regression analyses regarding muscle disorders had a significant beta weight of global stress that remained significant when controlling for the intervening variables age, gender and psychosomatic symptoms. The pattern of the stress-specific and unspecific CMD signs was consistent with the postulated stress model. Since there are positive results with regard to the stress model in patient samples and in this unselected sample of adolescents, further research is indicated, including the concept of somatization more explicitly.


Influence of actinomycin D, a RNA synthesis inhibitor, on long-term potentiation in rat hippocampal neurons in vivo and in vitro.
1. Hippocampal long-term potentiation (LTP) may serve as an elementary process underlying certain forms of learning and memory in vertebrates. As is the case with behavioural memory, hippocampal LTP in the rat CA1 region and in the dentate gyrus occurs in stages, which can be separated by an inhibitor of RNA synthesis. 2. Experiments have been performed in two brain regions, in the hippocampal CA1 region in vitro and in the dentate gyrus in vivo. 3. Maintenance of hippocampal LTP in the CA1 region in vitro was influenced by the RNA synthesis inhibitor actinomycin D from 4 h onwards. 4. The effect of actinomycin D on the time course of the population spike potentiation was more pronounced than the effect on the time course of the EPSP component, suggesting different mechanisms for the two forms of potentiation. 5. Intrahippocampal and intracerebroventricular injection of actinomycin D into rats prevented a late stage of LTP in the dentate gyrus in vivo measured as the population spike amplitude. 6. Since actinomycin D was only effective in influencing the maintenance of LTP when applied before tetanization, the requirement for transcription during LTP may have a critical time window. 7. Actinomycin D influenced the maintenance of LTP specifically, since the drug did not alter any potentials in control experiments after its removal or when it was administered shortly after tetanization. A second, structurally different RNA synthesis inhibitor, 5,6-dichloro-1-beta-D-ribofuranosyl benzimidazole, mimicked the effect of actinomycin D in vitro.


Screening for deep vein thrombosis and pulmonary embolism in outpatients with suspected DVT or PE by the sequential use of clinical score: a sensitive quantitative D-dimer test and noninvasive diagnostic tools.
The requirement for a safe diagnostic strategy should be based on an overall posttest incidence of venous thromboembolism (VTE) of less than 1% during 3-month follow-up. The negative predictive value (NPV) during 3 months of follow-up is 98.1 to 99% after a normal venogram, 97 to 98% after a normal compression ultrasonography (CUS), and > 99% after serial CUS testing. Serial CUS testing is safe but 100 CUS must be repeated to find one or two CUS positive for deep vein thrombosis (DVT), which is not cost-effective and indicates the need to improve the diagnostic workup of DVT by the use of clinical score assessment and D-dimer testing. The NPV varies from 97.6 to 99.4% for low clinical score followed by a negative SimpiRED test, indicating the need for a first CUS. The NPV is 98.4 to 99.3% for a normal rapid enzyme-linked immunosorbent assay (ELISA) VIDAS D-dimer test result (< 500 ng/mL) irrespective of clinical score. The NPV is more than 99% for a negative CUS followed by either a negative SimpiRED test or an ELISA VIDAS test result of < 1000 ng/mL without the need to repeat a second CUS within 1 week. The sequential use of a sensitive, rapid ELISA D-dimer and clinical score assessment will safely reduce the need for CUS testing by 40 to 60%. Large prospective outcome studies demonstrate that with one negative examination with complete duplex color ultrasonography (CCUS) of the proximal and distal veins of the affected leg with suspected DVT, it is safe to withhold anticoagulant treatment, with a negative predictive value of 99.5%. This may indicates that CCUS is equal to serial CUS or the combined use of clinical score, D-dimer testing, and CUS. Pulmonary angiography is the gold standard for segmental pulmonary embolism (PE) but not for subsegmental PE. A normal perfusion lung scan and a normal rapid ELISA VIDAS D-dimer test safely excludes PE. Helical spiral computed tomography (CT) detects all clinically relevant PE and a large number of alternative diagnoses in symptomatic patients with suspected PE and can replace both the ventilation perfusion scan and pulmonary angiography to safely rule in PE and to rule out PE with an NPV of > 99%. The combination of clinical assessment, a rapid ELISA VIDAS D-dimer, followed by CUS will reduce the need for helical spiral CT by 40 to 50%.


The determinants of home and nursing home death: a systematic review and meta-analysis.
Most Canadians die in hospital, and yet, many express a preference to die at home. Place of death is the result of the interaction among sociodemographic, illness- and healthcare-related factors. Although home death is sometimes considered a potential indicator of end-of-life/palliative care quality, some determinants of place of death are more modifiable than others. The objective of this systematic review was to evaluate the determinants of home and nursing home death in adult patients diagnosed with an advanced, life-limiting illness. A systematic literature search was performed for studies in English published from January 1, 2004 to September 24, 2013 that evaluated the determinants of home or nursing home death compared to hospital death in adult patients with an advanced, life-limiting condition. The adjusted odds ratios, relative risks, and 95% confidence intervals of each determinant were extracted from the studies. Meta-analyses were performed if appropriate. The quality of individual studies was assessed using the Newcastle-Ottawa scale and the body of evidence was assessed according to the GRADE Working Group criteria. Of the 5,900 citations identified, 26 retrospective cohort studies were eligible. The risk of bias in the studies identified was considered low. Factors associated with an increased likelihood of home versus hospital death included multidisciplinary home palliative care, preference for home death, cancer as opposed to other diagnoses, early referral to palliative care, not living alone, having a caregiver, and the caregiver's coping skills. Knowledge about the determinants of place of death can be used to inform care planning between healthcare providers, patients and family members regarding the feasibility of dying in the preferred location and may help explain the incongruence between preferred and actual place of death. Modifiable factors such as early referral to palliative care, presence of a multidisciplinary home palliative care team were identified, which may be amenable to interventions that improve the likelihood of a patient dying in the preferred location. Place of death may not be a very good indicator of the quality of end-of-life/palliative care since it is determined by multiple factors and is therefore dependent on individual circumstances.


Tongue traction is as effective as jaw lift maneuver for Trachlight-guided orotracheal intubation.
The jaw lift maneuver has been traditionally used to facilitate Trachlight-guided orotracheal intubation. The aim of this study was to compare the jaw lift maneuver with another potentially useful maneuver, tongue traction, for facilitating Trachlight-guided orotracheal intubation. This study was prospective and randomized in design. A single experienced investigator performed Trachlight-guided orotracheal intubations in 100 adult patients with clinically normal airways undergoing elective surgeries under general anesthesia with paralysis. Patients were divided into two groups: Group J (jaw lift); and Group T (tongue traction using a padded artery forceps). Three parameters were recorded: search time (device insertion to midline glow); time to intubate (device insertion to glow at suprasternal notch); and time to confirm (device insertion to confirmation of placement by capnography). All patients were intubated successfully using the Trachlight. Only one patient in Group J required more than two attempts for successful intubation. First and second attempt success rates were, respectively, 86% and 98% in Group J, and 92% and 100% in Group T. Mean search time, time to intubate and time to confirm were comparable: 6.36 ± 4.20 seconds, 11.23 ± 6.69 seconds and 21.59 ± 7.69 seconds, respectively, in Group J, and 6.81 ± 4.53 seconds, 10.79 ± 6.02 seconds and 22.80 ± 7.85 seconds, respectively, in Group T. The tongue traction maneuver is as good as the jaw lift maneuver with regard to the success rate and time taken for Trachlight-guided orotracheal intubation.


Outcomes of hydroxychloroquine usage in United States veterans hospitalized with Covid-19.
Despite limited and conflicting data on the use of hydroxychloroquine in patients with Covid-19, the U.S. Food and Drug Administration has authorized the emergency use of this drug when clinical trials are unavailable or infeasible. Hydroxychloroquine, alone or in combination with azithromycin, is being widely used in Covid-19 therapy based on anecdotal and limited observational evidence. We performed a retrospective analysis of data from patients hospitalized with confirmed SARS-CoV-2 infection in all United States Veterans Health Administration medical centers until April 11, 2020. Patients were categorized based on their exposure to hydroxychloroquine alone (HC) or with azithromycin (HC+AZ) as treatments in addition to standard supportive management for Covid-19. The two primary outcomes were death and the need for mechanical ventilation. We determined the association between treatment and the primary outcomes using competing risk hazard regression adjusting for clinical characteristics via propensity scores. Discharge and death were taken into account as competing risks and subdistribution hazard ratios are presented. A total of 368 patients were evaluated (HC, n=97; HC+AZ, n=113; no HC, n=158). Rates of death in the HC, HC+AZ, and no HC groups were 27.8%, 22.1%, 11.4%, respectively. Rates of ventilation in the HC, HC+AZ, and no HC groups were 13.3%, 6.9%, 14.1%, respectively. Compared to the no HC group, the risk of death from any cause was higher in the HC group (adjusted hazard ratio, 2.61; 95% CI, 1.10 to 6.17; P=0.03) but not in the HC+AZ group (adjusted hazard ratio, 1.14; 95% CI, 0.56 to 2.32; P=0.72). The risk of ventilation was similar in the HC group (adjusted hazard ratio, 1.43; 95% CI, 0.53 to 3.79; P=0.48) and in the HC+AZ group (adjusted hazard ratio, 0.43; 95% CI, 0.16 to 1.12; P=0.09), compared to the no HC group. In this study, we found no evidence that use of hydroxychloroquine, either with or without azithromycin, reduced the risk of mechanical ventilation in patients hospitalized with Covid-19. An association of increased overall mortality was identified in patients treated with hydroxychloroquine alone. These findings highlight the importance of awaiting the results of ongoing prospective, randomized, controlled studies before widespread adoption of these drugs.


Memory complaints in chronic pain.
In clinical practice, patients with chronic pain frequently report problems with memory functioning. This issue, however, has received little attention in the scientific literature. The present study was designed to investigate this common problem and to stimulate research interest in this neglected and important area. Self-reported memory problems were investigated in two groups of chronic pain patients--patients with pain from acceleration-deceleration automobile accidents (n = 56) and patients with pain from various work accidents (n = 27)--and two control groups involving medical/dental (n = 24) and psychotherapy patients (n = 20). Private practice, chronic pain, rehabilitation psychology services. Our findings suggest that memory complaints are higher in patients with chronic pain than in medical/dental or psychotherapy patients. No differences were found between chronic pain groups. On more general measures of memory complaint, differences between pain patients and controls were attributed to the severity of patients' depression. On a questionnaire designed to be more specific to memory complaint in chronic pain patients, differences in memory complaint between pain patients and controls were found, even after the effects due to depression were statistically removed. Although pain patients often attribute their memory problems to codeine use and/or psychoactive medications, there was no support for this in the present study. Within the limitations of this study, these findings suggest that memory complaints may be related not only to depression but also to the presence of chronic pain. Further research in this area is needed.


A cross sectional study of hepatitis B, C, some trace elements, heavy metals, aflatoxin B1 and schistosomiasis in a rural population, Egypt.
Chronic liver diseases are disastrous to health. Many factors are associated with their prevalence, hence endemicity. These are mainly infectious, parasitic and toxic. A survey was conducted in a village south to Cairo. Large industries concerned with iron and steel industry, metals smelting, cement manufacturing and electric station were located north to the village. A systematic random sample of houses was selected. All individuals inside the houses were invited to share in the study. Sample size was 84 individuals. Hepatitis markers were done (HBsAg and anti-HCV antibodies). The levels of some heavy metals were assessed; which were lead, mercury, arsenic, aluminum, manganese, nickel, chromium and cadmium. Levels of some trace elements were assessed. These were copper, iron, selenium and zinc. Aflatoxin B1 was assessed in serum. Assessment of schistosomal circulating antigen and antibodies was carried out. Abdominal ultrasonograghy was done to assess liver condition. Univariate logistic regression analysis was done to assess the association between studied variables and HBsAg or anti-HCV sero-positive subjects. The association between studied variables and bilharzial or fatty liver, diagnosed by ultrasonography, were also assessed. The univariate logistic regression analysis revealed odds ratios at the following results. For HBsAg seropositive subjects, aflatoxin B1, lead, chromium and schistosomal antigen and antibodies were higher than negative ones where odds ratios were; 6.2, 1.6, 1.6, 1.6 and 1.7, respectively. None of the variables showed statistically significant difference. For anti-HCV antibodies sero-positive subjects, aflatoxin B1 and chromium had the highest odds ratios among the studied variables, (odds ratios were 2.5 and 2.4, respectively). Bilharzial liver showed higher significant positivity of anti-HCV antibodies and insignificant decreased level of zinc than negative ones (odds ratios were 7.2 and 4.5, respectively). Fatty liver cases showed higher statistically significant positivity of anti-HCV antibodies and chromium than negative ones. Odds ratios were 8.0 and 7.1, respectively. Statistically significant lower level of aflatoxin B1 was shown in fatty liver than normal liver subjects. Multivariate logistic regression analysis for fatty liver showed that only anti-HCV antibodies sero-positivity had statistically significant odds ratio in comparison to chromium level and aflatoxin B1. It is concluded that some heavy metals, and Aflatoxin B1 had a definite association with liver diseases in the area under study. Having anti-HCV antibodies had a relation with fatty liver and with bilharzial liver more than having HBsAg. It is recommended that environmental management to factories nearby the village is urgently needed to decrease exposure to heavy metals. Prevention of hepatitis infection and aflatoxin exposure through different means is also recommended, other wise health care authorities would be confronted with unusual cases of HCC in the nearby future.


Thromboprophylaxis with fondaparinux in high-risk postoperative patients with renal insufficiency.
Fondaparinux is an antithrombin-dependent factor Xa inhibitor that is used for thromboprophylaxis of patients undergoing hip fracture surgery, hip or knee replacement, or abdominal surgery. It is cleared by the kidney and should be used with caution in patients with renal impairment and avoided in patients with severe renal insufficiency. Recently, several studies have demonstrated that a lower dose of fondaparinux in patients with moderate renal impairment appears to be safe and effective. The purpose of this study was to obtain pharmacokinetic and clinical data on the use of prophylactic fondaparinux in patients with renal insufficiency undergoing major abdominal surgery for cancer (n=8) or orthopedic surgery (n=1). Anti-factor Xa levels were obtained, and a published population pharmacokinetic model for fondaparinux was fit to the data. The data were analyzed using NONMEM software. Fondaparinux did not appear to accumulate in these patients, even when the drug was administered for up to twelve days. Pharmacokinetic analysis revealed that the apparent clearance in this population, who were primarily undergoing cancer surgery, was similar to prior studies in orthopedic surgery patients. In contrast, lower estimates were obtained for volume of distribution and absorption rate constant parameters. None of the patients sustained a hemorrhagic complication attributable to fondaparinux. One patient developed hypoxia in the setting of transient atrial fibrillation and clinical suspicion for pulmonary embolism, but this was not confirmed radiographically. These results support the use of 1.5mg of fondaparinux every 24hours for thromboprophylaxis in patients with renal insufficiency undergoing high-risk surgical procedures.


Functional triplet motifs underlie accurate predictions of single-trial responses in populations of tuned and untuned V1 neurons.
Visual stimuli evoke activity in visual cortical neuronal populations. Neuronal activity can be selectively modulated by particular visual stimulus parameters, such as the direction of a moving bar of light, resulting in well-defined trial averaged tuning properties. However, given any single stimulus parameter, a large number of neurons in visual cortex remain unmodulated, and the role of this untuned population is not well understood. Here, we use two-photon calcium imaging to record, in an unbiased manner, from large populations of layer 2/3 excitatory neurons in mouse primary visual cortex to describe co-varying activity on single trials in neuronal populations consisting of both tuned and untuned neurons. Specifically, we summarize pairwise covariability with an asymmetric partial correlation coefficient, allowing us to analyze the resultant population correlation structure, or functional network, with graph theory. Using the graph neighbors of a neuron, we find that the local population, including both tuned and untuned neurons, are able to predict individual neuron activity on a moment to moment basis, while also recapitulating tuning properties of tuned neurons. Variance explained in total population activity scales with the number of neurons imaged, demonstrating larger sample sizes are required to fully capture local network interactions. We also find that a specific functional triplet motif in the graph results in the best predictions, suggesting a signature of informative correlations in these populations. In summary, we show that unbiased sampling of the local population can explain single trial response variability as well as trial-averaged tuning properties in V1, and the ability to predict responses is tied to the occurrence of a functional triplet motif.


Saturation of the turbulent dynamo.
The origin of strong magnetic fields in the Universe can be explained by amplifying weak seed fields via turbulent motions on small spatial scales and subsequently transporting the magnetic energy to larger scales. This process is known as the turbulent dynamo and depends on the properties of turbulence, i.e., on the hydrodynamical Reynolds number and the compressibility of the gas, and on the magnetic diffusivity. While we know the growth rate of the magnetic energy in the linear regime, the saturation level, i.e., the ratio of magnetic energy to turbulent kinetic energy that can be reached, is not known from analytical calculations. In this paper we present a scale-dependent saturation model based on an effective turbulent resistivity which is determined by the turnover time scale of turbulent eddies and the magnetic energy density. The magnetic resistivity increases compared to the Spitzer value and the effective scale on which the magnetic energy spectrum is at its maximum moves to larger spatial scales. This process ends when the peak reaches a characteristic wave number k☆ which is determined by the critical magnetic Reynolds number. The saturation level of the dynamo also depends on the type of turbulence and differs for the limits of large and small magnetic Prandtl numbers Pm. With our model we find saturation levels between 43.8% and 1.3% for Pm≫1 and between 2.43% and 0.135% for Pm≪1, where the higher values refer to incompressible turbulence and the lower ones to highly compressible turbulence.


An in vitro comparison of effect on fracture strength, pH and calcium ion diffusion from various biomimetic materials when used for repair of simulated root resorption defects.
The aim of this study was to compare the effect on fracture strength, pH and calcium ion diffusion from mineral trioxide aggregate (MTA) Fillapex, iRoot SP, and Ultracal when used for repair of simulated root resorption defects. Three sets of 40 teeth each were used, and biomechanical preparation was done. Resorption cavity was made at 5 mm from the apex. Teeth were filled with different experimental materials. In control group, saline was used. Samples of fracture resistance were stored in an incubator for 3 months and then subjected to the universal testing machine. To evaluate the pH and calcium ion release were checked at 1 day and 1, 2, 3, and 4 weeks intervals. Data were recorded and statistical analysis done by one-way analysis of variance followed by post-hoc Tukey test. Highest fracture resistance was seen in MTA Fillapex followed by iRoot SP, control, and then Ultracal group. Teeth filled with iRoot SP showed highest pH and calcium ion release followed by MTA Fillapex and Ultracal group. Bioceramic sealers showed high pH, calcium ion release, and good root reinforcement potential. Initial dressing of calcium hydroxide followed by obturation with Gutta-percha and bioceramic sealer may be considered as an alternative treatment modality for inflammatory resorption.


Self-assembly of like-charged nanoparticles into microscopic crystals.
Like-charged nanoparticles, NPs, can assemble in water into large, faceted crystals, each made of several million particles. These NPs are functionalized with mixed monolayers comprising ligands terminating in carboxylic acid group ligands as well as positively charged quaternary ammonium ligands. The latter groups give rise to electrostatic interparticle repulsions which partly offset the hydrogen bonding between the carboxylic acids. It is the balance between these two interactions that ultimately enables self-assembly. Depending on the pH, the particles can crystallize, form aggregates, remain unaggregated or even - in mixtures of two particle types - can "choose" whether to crystallize with like-charged or oppositely charged particles.


Microbial metalloproteomes are largely uncharacterized.
Metal ion cofactors afford proteins virtually unlimited catalytic potential, enable electron transfer reactions and have a great impact on protein stability. Consequently, metalloproteins have key roles in most biological processes, including respiration (iron and copper), photosynthesis (manganese) and drug metabolism (iron). Yet, predicting from genome sequence the numbers and types of metal an organism assimilates from its environment or uses in its metalloproteome is currently impossible because metal coordination sites are diverse and poorly recognized. We present here a robust, metal-based approach to determine all metals an organism assimilates and identify its metalloproteins on a genome-wide scale. This shifts the focus from classical protein-based purification to metal-based identification and purification by liquid chromatography, high-throughput tandem mass spectrometry (HT-MS/MS) and inductively coupled plasma mass spectrometry (ICP-MS) to characterize cytoplasmic metalloproteins from an exemplary microorganism (Pyrococcus furiosus). Of 343 metal peaks in chromatography fractions, 158 did not match any predicted metalloprotein. Unassigned peaks included metals known to be used (cobalt, iron, nickel, tungsten and zinc; 83 peaks) plus metals the organism was not thought to assimilate (lead, manganese, molybdenum, uranium and vanadium; 75 peaks). Purification of eight of 158 unexpected metal peaks yielded four novel nickel- and molybdenum-containing proteins, whereas four purified proteins contained sub-stoichiometric amounts of misincorporated lead and uranium. Analyses of two additional microorganisms (Escherichia coli and Sulfolobus solfataricus) revealed species-specific assimilation of yet more unexpected metals. Metalloproteomes are therefore much more extensive and diverse than previously recognized, and promise to provide key insights for cell biology, microbial growth and toxicity mechanisms.


Unapparent foot and mouth disease infection (sub-clinical infections and carriers): implications for control.
Unlike animals which are carriers of foot and mouth disease (FMD), sub-clinically infected animals may be highly contagious. The implications of sub-clinical infections for the control of FMD are serious because such animals are likely to disseminate the disease when in contact with susceptible livestock. Recent dissemination of FMD virus (FMDV) in Europe shows that sub-clinically infected animals render trade in animals or animal products a potential risk for importing countries. This clearly demonstrates that the paradigm 'free of FMD without vaccination' is not synonymous with 'risk-free'. The risk of introduction of subclinical FMD into FMD-free countries may increase significantly, with the occurrence of large susceptible animal populations, changed agricultural practices, expansion of trade in live animals and animal movements, increased trade in animal products and greater mobility of people. Such changes in circumstances require that national and international authorities remain continuously vigilant to determine any altered risk for importation of FMD. A few historical reports and some recent observations in southern Africa indicate the possibility of dissemination of FMD by bovine carriers into herds of susceptible cattle. These reports have greatly influenced FMD trade policies and thus, FMD control and eradication strategies. However, other field evidence does not support this claim and several controlled experiments were unable to show that carriers are able to initiate disease. When millions of cattle were systematically vaccinated with good quality vaccines, FMD disappeared in spite of a large sentinel population in the form of calves and unvaccinated sheep and pigs. A low number of carriers most likely persisted, but they did not hamper the eradication of the disease. Vaccination policies and trade regulation must be based on risk assessments taking these factors into consideration.


A synthesis of convergent reflections, tensions and silences in linking gender and global environmental change research.
This synthesis article joins the authors of the special issue "Gender perspectives in resilience, vulnerability and adaptation to global environmental change" in a common reflective dialogue about the main contributions of their papers. In sum, here we reflect on links between gender and feminist approaches to research in adaptation and resilience in global environmental change (GEC). The main theoretical contributions of this special issue are threefold: emphasizing the relevance of power relations in feminist political ecology, bringing the livelihood and intersectionality approaches into GEC, and linking resilience theories and critical feminist research. Empirical insights on key debates in GEC studies are also highlighted from the nine cases analysed, from Europe, the Americas, Asia, Africa and the Pacific. Further, the special issue also contributes to broaden the gender approach in adaptation to GEC by incorporating research sites in the Global North alongside sites from the Global South. This paper examines and compares the main approaches adopted (e.g. qualitative or mixed methods) and the methodological challenges that derive from intersectional perspectives. Finally, key messages for policy agendas and further research are drawn from the common reflection.


Sequential morphologic events during apoptosis of human neutrophils. Modulation by lipoxygenase-derived eicosanoids.
Dual-laser flow cytometry, based on the properties of the DNA-binding dyes Hoechst 33342 and propidium iodide, was used, with light and electron microscopy and DNA fragmentation studies, to define the influence of lipoxygenase-derived eicosanoids on apoptosis of human polymorphonuclear neutrophils (PMN) in vitro. Apoptosis was characterized by progression through an early apoptotic phase characterized by condensation of chromatin and coalescence of nuclear lobes, to a late apoptotic phase characterized by nuclear degradation and evanescence, and secondary necrosis. Prolonged exposure of PMN to leukotriene B4 (LTB4) afforded dose-dependent inhibition of constitutive PMN apoptosis (percentage of normal and apoptotic PMN, respectively, after aging for 18 h: vehicle, 30.5 +/- 2.7% and 61.8 +/- 3.2%; LTB4 10(-7) M, 57.6 +/- 1.2% and 37.6 +/- 1.0%) and apoptosis triggered by the classic peptide chemoattractant FMLP. In contrast, apoptosis was not affected by the LTB4 precursor 5(S)-hydroxyeicosatetraenoic acid (HETE), the omega-oxidation LTB4 metabolites 20-hydroxy-LTB4 and 20-carboxy-LTB4, the cysteinyl leukotriene LTC4, the 15-lipoxygenase product 15(S)-HETE, or the lipoxygenase interaction product lipoxin A4. The anti-apoptotic effect of LTB4 was mimicked by 20,20,20-trifluoro-LTB4, LTB4-dimethylamide, and 14,15-dehydro-LTB4, and was blunted by pertussis toxin and genistein, inhibitors of G alpha i GTP-binding proteins and tyrosine kinases, respectively, but not by staurosporine, 15(S)-HETE, or lipoxin A4. This unique pharmacologic profile suggested that LTB4 attenuated apoptosis through activation of cell surface receptors and signaling events distinct from those involved with PMN trafficking, degranulation, and respiratory bursts.


Social concepts are represented in the superior anterior temporal cortex.
Social concepts such as "tactless" or "honorable" enable us to describe our own as well as others' social behaviors. The prevailing view is that this abstract social semantic knowledge is mainly subserved by the same medial prefrontal regions that are considered essential for mental state attribution and self-reflection. Nevertheless, neurodegeneration of the anterior temporal cortex typically leads to impairments of social behavior as well as general conceptual knowledge. By using functional MRI, we demonstrate that the anterior temporal lobe represents abstract social semantic knowledge in agreement with this patient evidence. The bilateral superior anterior temporal lobes (Brodmann's area 38) are selectively activated when participants judge the meaning relatedness of social concepts (e.g., honor-brave) as compared with concepts describing general animal functions (e.g., nutritious-useful). Remarkably, only activity in the superior anterior temporal cortex, but not the medial prefrontal cortex, correlates with the richness of detail with which social concepts describe social behavior. Furthermore, this anterior temporal lobe activation is independent of emotional valence, whereas medial prefrontal regions show enhanced activation for positive social concepts. Our results demonstrate that the superior anterior temporal cortex plays a key role in social cognition by providing abstract conceptual knowledge of social behaviors. We further speculate that these abstract conceptual representations can be associated with different contexts of social actions and emotions through integration with frontolimbic circuits to enable flexible evaluations of social behavior.


MKP-7, a novel mitogen-activated protein kinase phosphatase, functions as a shuttle protein.
Mitogen-activated protein kinase (MAPK) phosphatases (MKPs) negatively regulate MAPK activity. In the present study, we have identified a novel MKP, designated MKP-7, and mapped it to human chromosome 12p12. MKP-7 possesses a long C-terminal stretch containing both a nuclear export signal and a nuclear localization signal, in addition to the rhodanese-like domain and the dual specificity phosphatase catalytic domain, both of which are conserved among MKP family members. When expressed in mammalian cells MKP-7 protein was localized exclusively in the cytoplasm, but this localization became exclusively nuclear following leptomycin B treatment or introduction of a mutation in the nuclear export signal. These findings indicate that MKP-7 is the first identified leptomycin B-sensitive shuttle MKP. Forced expression of MKP-7 suppressed activation of MAPKs in COS-7 cells in the order of selectivity, JNK p38 > ERK. Furthermore, a mutant form MKP-7 functioned as a dominant negative particularly against the dephosphorylation of JNK, suggesting that MKP-7 works as a JNK-specific phosphatase in vivo. Co-immunoprecipitation experiments and histological analysis suggested that MKP-7 determines the localization of MAPKs in the cytoplasm.


Nuclear targeting of dystroglycan promotes the expression of androgen regulated transcription factors in prostate cancer.
Dystroglycan is frequently lost in adenocarcinoma, but the mechanisms and consequences are poorly understood. We report an analysis of β-dystroglycan in prostate cancer in human tissue samples and in LNCaP cells in vitro. There is progressive loss of β-dystroglycan immunoreactivity from basal and lateral surfaces of prostate epithelia which correlates significantly with increasing Gleason grade. In about half of matched bone metastases there is significant dystroglycan re-expression. In tumour tissue and in LNCaP cells there is also a tyrosine phosphorylation-dependent translocation of β-dystroglycan to the nucleus. Analysis of gene expression data by microarray, reveals that nuclear targeting of β-dystroglycan in LNCaP cells alters the transcription of relatively few genes, the most unregulated being the transcription factor ETV1. These data suggest that proteolysis, tyrosine phosphorylation and translocation of dystroglycan to the nucleus resulting in altered gene transcription could be important mechanisms in the progression of prostate cancer.


The agmR gene, an environmentally responsive gene, complements defective glpR, which encodes the putative activator for glycerol metabolism in Pseudomonas aeruginosa.
The genes for the peripheral glycerol carbon metabolic pathway (glp) in Pseudomonas aeruginosa are postulated to be positively regulated by GlpR. A gene complementing the glpR2 allele, affecting expression of the putative activator, was cloned by a bacteriophage mini-D3112-based in vivo cloning method. Mini-D3112 replicons were isolated by transfecting glpR2 strain PRP406 and selecting clones able to grow on minimal medium containing glycerol as the sole carbon and energy source. Preliminary biochemical characterization indicated that the cloned activator gene for glycerol metabolism (agmR) may not be allelic to glpR. Restriction analysis and recloning of DNA fragments located the agmR gene to a 2.3-kb EcoRV-SstI DNA fragment. In a T7 RNA polymerase expression system, a single 26,000-Da protein was expressed from this DNA fragment. The amino acid sequence of this protein, deduced from the nucleotide sequence reported here, demonstrates its homology to the effector (or regulator) proteins of the environmentally responsive two-component regulators. The carboxy-terminal region of AgmR contains a possible helix-turn-helix DNA-binding motif and resembles sequences found in transcriptional regulators of the LuxR family.


Cryptic footprints of rare earth elements on natural resources and living organisms.
Rare earth elements (REEs) are gaining attention due to rapid rise of modern industries and technological developments in their usage and residual fingerprinting. Cryptic entry of REEs in the natural resources and environment is significant; therefore, life on earth is prone to their nasty effects. Scientific sectors have expressed concerns over the entry of REEs into food chains, which ultimately influences their intake and metabolism in the living organisms. Extensive scientific collections and intensive look in to the latest explorations agglomerated in this document aim to depict the distribution of REEs in soil, sediments, surface waters and groundwater possibly around the globe. Furthermore, it draws attention towards potential risks of intensive industrialization and modern agriculture to the exposure of REEs, and their effects on living organisms. It also draws links of REEs usage and their footprints in natural resources with the major food chains involving plants, animals and humans. Scientific literature preferably spanning over the last five years was obtained online from the MEDLINE and other sources publishing the latest studies on REEs distribution, properties, usage, cycling and intrusion in the environment and food-chains. Distribution of REEs in agricultural soils, sediments, surface and ground water was drawn on the global map, together with transport pathways of REEs and their cycling in the natural resources. Fourteen REEs (Ce, Dy, Er, Eu, Gd, Ho, La, Lu, Nd, Pr, Sm, Tb, Th and Yb) were plighted in this study. Wide range of their concentrations has been detected in agricultural soils (<15.9-249.1 μg g-1) and in groundwater (<3.1-146.2 μg L-1) at various sites worldwide. They have strong tendency to accumulate in the human body, and thus associated with kidney stones. The REEs could also perturb the animal physiology, especially affecting the reproductive development in both terrestrial and aquatic animals. In plants, REEs might affect the germination, root and shoot development and flowering at concentration ranging from 0.4 to 150 mg kg-1. This review article precisely narrates the current status, sources, and potential effects of REEs on plants, animals, humans health. There are also a few examples where REEs have been used to benefit human health. However, still there is scarce information about threshold levels of REEs in the soil, aquatic, and terrestrial resources as well as living entities. Therefore, an aggressive effort is required for global action to generate more data on REEs. This implies we prescribe an urgent need for inter-disciplinary studies about REEs in order to identify their toxic effects on both ecosystems and organisms.


piggybac- and PhiC31-mediated genetic transformation of the Asian tiger mosquito, Aedes albopictus (Skuse).
The Asian tiger mosquito, Aedes albopictus (Skuse), is a vector of several arboviruses including dengue and chikungunya. This highly invasive species originating from Southeast Asia has travelled the world in the last 30 years and is now established in Europe, North and South America, Africa, the Middle East and the Caribbean. In the absence of vaccine or antiviral drugs, efficient mosquito control strategies are crucial. Conventional control methods have so far failed to control Ae. albopictus adequately. Germline transformation of Aedes albopictus was achieved by micro-injection of embryos with a piggyBac-based transgene carrying a 3xP3-ECFP marker and an attP site, combined with piggyBac transposase mRNA and piggyBac helper plasmid. Five independent transgenic lines were established, corresponding to an estimated transformation efficiency of 2-3%. Three lines were re-injected with a second-phase plasmid carrying an attB site and a 3xP3-DsRed2 marker, combined with PhiC31 integrase mRNA. Successful site-specific integration was observed in all three lines with an estimated transformation efficiency of 2-6%. Both piggybac- and site-specific PhiC31-mediated germline transformation of Aedes albopictus were successfully achieved. This is the first report of Ae. albopictus germline transformation and engineering, a key step towards studying and controlling this species using novel molecular techniques and genetic control strategies.


Analysis of corneal and conjunctival microenvironments using monoclonal antibodies.
The authors phenotypically compared epithelial and nonepithelial components of human corneal and conjunctival microenvironments using a panel of monoclonal antibody reagents that included markers of epithelial cell maturation, markers of mesodermal-derived fibrous tissue and vessels, markers of specific keratins, and markers of major histocompatibility complex Class I and II antigens. Corneoscleral rims obtained after trephination of the donor button for use in penetrating keratoplasty procedures were studied. A comparison of cornea and conjunctiva with anti-epithelial and antikeratin antibodies demonstrated that corneal and conjunctival keratinocytes undergo similar antigenically defined pathways of maturation. However, the reactivity of antibody 12/1-2 (antibody against low molecular weight keratins) with conjunctival but not corneal basal cells suggested differences in keratin expression between the two epithelial types. Using antibodies against major histocompatibility complex Class I and II antigens, it was demonstrated the two tissues were similar with Class I determinants found on all epithelial, stromal, and endothelial cells, and Class II determinants found on Langerhans' cells, vessels, and a subset of stromal cells. The availability of tissue-specific markers of epithelial and nonepithelial components of the cornea and conjunctiva should be of use in the study of the roles the ocular microenvironment might play in the pathogenesis of ocular inflammatory diseases.


Loss of MARCH5 mitochondrial E3 ubiquitin ligase induces cellular senescence through dynamin-related protein 1 and mitofusin 1.
Mitochondria constantly divide and combine through fission and fusion activities. MARCH5, a mitochondrial E3 ubiquitin ligase, has been identified as a molecule that binds mitochondrial fission 1 protein (hFis1), dynamin-related protein 1 (Drp1) and mitofusin 2 (Mfn2), key proteins in the control of mitochondrial fission and fusion. However, how these interactions control mitochondrial dynamics, and cellular function has remained obscure. Here, we show that shRNA-mediated MARCH5 knockdown promoted the accumulation of highly interconnected and elongated mitochondria. Cells transfected with MARCH5 shRNA or a MARCH5 RING domain mutant displayed cellular enlargement and flattening accompanied by increased senescence-associated beta-galactosidase (SA-beta-Gal) activity, indicating that these cells had undergone cellular senescence. Notably, a significant increase in Mfn1 level, but not Mfn2, Drp1 or hFis1 levels, was observed in MARCH5-depleted cells, indicating that Mfn1 is a major ubiquitylation substrate. Introduction of Mfn1(T109A), a GTPase-deficient mutant form of Mfn1, into MARCH5-RNAi cells not only disrupted mitochondrial elongation, but also abolished the increase in SA-beta-Gal activity. Moreover, the aberrant mitochondrial phenotypes in MARCH5-RNAi cells were reversed by ectopic expression of Drp1, but not by hFis1, and reversion of the mitochondria morphology in MARCH5-depleted cells was accompanied by a reduction in SA-beta-Gal activity. Collectively, our data indicate that the lack of MARCH5 results in mitochondrial elongation, which promotes cellular senescence by blocking Drp1 activity and/or promoting accumulation of Mfn1 at the mitochondria.