Characterization of the diversity and temporal stability of bacterial communities in human milk.
Recent investigations have demonstrated that human milk contains a variety of bacterial genera; however, as of yet very little work has been done to characterize the full diversity of these milk bacterial communities and their relative stability over time. To more thoroughly investigate the human milk microbiome, we utilized microbial identification techniques based on pyrosequencing of the 16S ribosomal RNA gene. Specifically, we characterized the bacterial communities present in milk samples collected from 16 women at three time-points over four weeks. Results indicated that milk bacterial communities were generally complex; several genera represented greater than 5% of the relative community abundance, and the community was often, yet not always, stable over time within an individual. These results support the conclusion that human milk, which is recommended as the optimal nutrition source for almost all healthy infants, contains a collection of bacteria more diverse than previously reported. This finding begs the question as to what role this community plays in colonization of the infant gastrointestinal tract and maintaining mammary health.


State Variability in Diagnosed Conditions for IDEA Part C Eligibility.
An infant or toddler can begin the process of receiving Part C early intervention services by having a diagnosed condition with a high probability of developmental delay (Individuals with Disabilities Education Improvement Act, 2004). How states define those diagnosed conditions that begin the initiation process varies widely. Lists of diagnosed conditions were collected from state Part C websites and Part C coordinators for a descriptive analysis. Across 49 states, the District of Columbia, and 4 territories, a final list of 620 unique conditions was compiled. No single condition was listed by all jurisdictions. Hearing impairment was the condition listed by the most states (n = 38), followed by fetal alcohol syndrome (n = 34). Of the 620 conditions, 168 (27%) were listed by only 1 state, 554 (89%) were listed by fewer than 10 states, and 66 (11%) were listed by 10 or more states. Of these 66 conditions, 47 (71%) were listed by fewer than 20 states. Most of these 66 conditions (n = 48; 72.7%) had a prevalence of "very rare or rare," 8 (12%) were "common," 6 (9%) were "very common," and 4 (6.1%) were "unknown." The wide heterogeneity in the number and type of diagnostic conditions listed across states should be further investigated as it may represent imbalances in children with diagnosed conditions gaining access to Part C evaluations and individualized family service plans and potentially the services themselves across states. In addition, providing ready access to lists of diagnosed conditions is a simple step that could help states and Part C programs facilitate access to services.


Adherence to recommended exercise guidelines in patients with heart failure.
Exercise training has been shown to be beneficial in patients with heart failure, and its effectiveness is connected to adherence to the exercise program. Nonetheless, adherence to exercise in these patients remains a concern. Heart failure patients can be considered adherent to an exercise program if they meet 80 % of the recommended dose. We summarize exercise recommendations for patients with heart failure, identify exercise prescription methodologies used in studies that have reported exercise adherence, identify strategies and tools used to improve adherence and examine whether these strategies were developed using a theoretical platform with the primary aim to change behavior and improve adherence to exercise. Factors which may also impact adherence such as exercise setting, intensity and length of participation, gender, race, New York Heart Association functional class and heart failure with preserved and reduced ejection fraction were also investigated. Finally, recommendations for future studies for improving adherence to exercise in patients with heart failure are provided.


Branched-chain amino acid-enriched supplementation improves insulin resistance in patients with chronic liver disease.
Increased insulin resistance is a therapeutic target in patients with chronic liver disease. Branched-chain amino acids (BCAA) have been reported to improve insulin resistance in in vivo experiments. Thus, we investigated the effects of BCAA on insulin resistance in patients with chronic liver disease. Twelve patients with chronic liver disease were enrolled. Each patient was given one sachet of a BCAA-enriched supplement after breakfast and another at bedtime. The effects of the BCAA-enriched supplementation on insulin resistance were examined 30, 60 and 90 days after administration by the homeostasis model assessment method for insulin resistance (HOMA-IR) and for beta cell function (HOMA-%B). The HOMA-IR and HOMA-%B values were elevated at baseline, however, these parameters showed no significant changes after administration of the BCAA-enriched supplement in the overall patient population. By stratification via gender, patients in the male group showed a significantly greater elevation in the HOMA-IR value compared to the female patients at baseline. After the administration, the HOMA-IR and HOMA-%B values were significantly decreased only in the male group (9.4+/-4.8 vs. 2.4+/-0.7, 657+/-345 vs. 126+/-36, respectively; P<0.05). We found that there was a gender difference in chronic viral liver disease-related insulin resistance. Moreover, a BCAA-enriched supplement improved insulin resistance and beta cell function in male patients with chronic viral liver disease. Thus, a BCAA-enriched supplement may be a useful therapeutic agent for decreasing insulin resistance in male patients with chronic viral liver disease.


Gene editing of human embryos is not contrary to human rights law: A reply to Drabiak.
In an article in this journal, Katherine Drabiak argues that green lighting genome editing of human embryos is contrary to "fundamental human rights law." According to the author, genome editing of human embryos violates what we should recognize as a fundamental human right to inherit a genome without deliberate manipulation. In this reply article, we assess Drabiak's legal analysis and show methodological and substantive flaws. Methodologically, her analysis omits the key international legal instruments that form the so-called International Bill of Rights and thus fails to provide a full and accurate account of the fundamental international human rights standards. Substantively, Drabiak invokes, as a basis for prohibiting gene editing of human embryos, a legal standard (the rights and integrity of the future child) that is unknown to international law. Contrary to Drabiak's account, genome editing of human embryos is not prohibited under international law. Indeed, the right to health and the right to benefit from scientific progress may be interpreted as the basis of a legal duty to provide equality of access to germline gene editing, once determined it is beneficial and safe to use.


Luminal signalling links cell communication to tissue architecture during organogenesis.
Morphogenesis is the process whereby cell collectives are shaped into differentiated tissues and organs. The self-organizing nature of morphogenesis has been recently demonstrated by studies showing that stem cells in three-dimensional culture can generate complex organoids, such as mini-guts, optic-cups and even mini-brains. To achieve this, cell collectives must regulate the activity of secreted signalling molecules that control cell differentiation, presumably through the self-assembly of microenvironments or niches. However, mechanisms that allow changes in tissue architecture to feedback directly on the activity of extracellular signals have not been described. Here we investigate how the process of tissue assembly controls signalling activity during organogenesis in vivo, using the migrating zebrafish lateral line primordium. We show that fibroblast growth factor (FGF) activity within the tissue controls the frequency at which it deposits rosette-like mechanosensory organs. Live imaging reveals that FGF becomes specifically concentrated in microluminal structures that assemble at the centre of these organs and spatially constrain its signalling activity. Genetic inhibition of microlumen assembly and laser micropuncture experiments demonstrate that microlumina increase signalling responses in participating cells, thus allowing FGF to coordinate the migratory behaviour of cell groups at the tissue rear. As the formation of a central lumen is a self-organizing property of many cell types, such as epithelia and embryonic stem cells, luminal signalling provides a potentially general mechanism to locally restrict, coordinate and enhance cell communication within tissues.


Stromal vascular fraction-enriched fat grafting as treatment of adherent scars: study design of a non-randomized early phase trial.
In the last decades, autologous fat grafting has been used to treat adherent dermal scars. The observed regenerative and scar-reducing properties have been mainly ascribed to the tissue-derived stromal vascular fraction (tSVF) in adipose tissue. Adipose tissue's components augment local angiogenesis and mitosis in resident tissue cells. Moreover, it promotes collagen remodeling. We hypothesize that tSVF potentiates fat grafting-based treatment of adherent scars. Therefore, this study aims to investigate the effect of tSVF-enriched fat grafting on scar pliability over a 12-month period. A clinical multicenter non-randomized early phase trial will be conducted in two dedicated Dutch Burn Centers (Red Cross Hospital, Beverwijk, and Martini Hospital, Groningen). After informed consent, 46 patients (≥18 years) with adherent scars caused by burns, necrotic fasciitis, or degloving injury who have an indication for fat grafting will receive a sub-cicatricic tSVF-enriched fat graft. The primary outcome is the change in scar pliability measured by the Cutometer between pre- and 12 months post-grafting. Secondary outcomes are scar pliability (after 3 months), scar erythema, and melanin measured by the DSM II Colormeter; scar quality assessed by the patient and observer scales of the Patient and Observer Scar Assessment Scale (POSAS) 2.0; and histological analysis of scar biopsies (voluntary) and tSVF quality and composition. This study has been approved by the Dutch Central Committee for Clinical Research (CCMO), NL72094.000.20. This study will test the clinical efficacy of tSVF-enriched fat grafting to treat dermal scars while the underlying working mechanism will be probed into too. Dutch Trial Register NL 8461. Registered on 16 March 2020.


Fluoride supplements for young children: an analysis of the literature focusing on benefits and risks.
The use of fluoride supplements to prevent caries has been advised for more than 100 years, but serious promotion of this strategy occurred only after the effectiveness of water fluoridation was established, in the late 1950s and 1960s. Although the effectiveness of fluoride supplements was apparently endorsed by many small clinical studies, closer examination of the experimental conditions of these, their methods and the analysis of their results undermined confidence in their findings. It is likely that confounding resulted in spurious conclusions in many of them. More modern, well-conducted clinical trials of supplements suggest that today, in children also exposed to fluoride from other sources such as toothpaste, the marginal effect of fluoride supplements is very small. There is evidence that fluoride lozenges, designed to maximise any local effect, may have a small caries preventive effect, particularly in deciduous teeth. Overall, poor compliance makes fluoride supplements a poor public health measure. Supplement use by young children is associated with a substantial risk of dental fluorosis. Fluorosis is an issue about which the public is becoming concerned in several countries and this concern, if translated into opposition to all fluoride use, could jeopardise the most successful caries preventive aid we have. The potential for dental fluorosis, concern about the public's reaction to this, the poor effectiveness of supplements and the public's poor compliance with their use are persuasive arguments for a radical reduction in the use of supplements by young children. Recent changes in fluoride dosage schedules and deferment of the age of commencing the use of supplements, implemented in many countries, have followed from these concerns. Supplements formulated as lozenges maximise topical exposure of enamel to fluoride and such products may offer older children and some adults a way of maintaining an elevated fluoride level in saliva at times when toothbrushing is not practical.


Preference for wine is associated with lower hip fracture incidence in post-menopausal women.
Past studies of relationships between alcohol and hip fracture have generally focused on total alcohol consumed and not type of alcohol. Different types of alcohol consist of varying components which may affect risk of hip fracture differentially. This study seeks to examine the relationship between alcohol consumption, with a focus on type of alcohol consumed (e.g. beer, wine, or hard liquor) and hip fracture risk in post-menopausal women. The longitudinal cohort consisted of U.S. post-menopausal women aged 50-79 years enrolled between 1993-1998 in the Women's Health Initiative Clinical Trials and Observational Study (N=115,655). Women were categorized as non-drinkers, past drinkers, infrequent drinkers and drinkers by preference of alcohol type (i.e. those who preferred wine, beer, hard liquor, or who had no strong preference). Mean alcohol consumption among current drinkers was 3.3 servings per week; this was similar among those who preferred wine, beer and liquor. After adjustment for potential confounders, alcohol preference was strongly correlated with hip fracture risk (p = 0.0167); in particular, women who preferred wine were at lower risk than non-drinkers (OR=0.78; 95% CI 0.64-0.95), past drinkers (OR=0.85; 95% CI 0.72-1.00), infrequent drinkers (OR=0.73; 95% CI 0.61-0.88), hard liquor drinkers (OR=0.87; 95% CI 0.71-1.06), beer drinkers (OR=0.72; 95% CI 0.55-0.95) and those with no strong preference (OR=0.89; 95% CI 0.89; 95% CI 0.73-1.10). Preference of alcohol type was associated with hip fracture; women who preferentially consumed wine had a lower risk of hip fracture compared to non-drinkers, past drinkers, and those with other alcohol preferences.


Impact of violated high-dose refuge assumptions on evolution of Bt resistance.
Transgenic crops expressing Bacillus thuringiensis (Bt) toxins have been widely and successfully deployed for the control of target pests, while allowing a substantial reduction in insecticide use. The evolution of resistance (a heritable decrease in susceptibility to Bt toxins) can pose a threat to sustained control of target pests, but a high-dose refuge (HDR) management strategy has been key to delaying countervailing evolution of Bt resistance. The HDR strategy relies on the mating frequency between susceptible and resistant individuals, so either partial dominance of resistant alleles or nonrandom mating in the pest population itself could elevate the pace of resistance evolution. Using classic Wright-Fisher genetic models, we investigated the impact of deviations from standard refuge model assumptions on resistance evolution in the pest populations. We show that when Bt selection is strong, even deviations from random mating and/or strictly recessive resistance that are below the threshold of detection can yield dramatic increases in the pace of resistance evolution. Resistance evolution is hastened whenever the order of magnitude of model violations exceeds the initial frequency of resistant alleles. We also show that the existence of a fitness cost for resistant individuals on the refuge crop cannot easily overcome the effect of violated HDR assumptions. We propose a parametrically explicit framework that enables both comparison of various field situations and model inference. Using this model, we propose novel empiric estimators of the pace of resistance evolution (and time to loss of control), whose simple calculation relies on the observed change in resistance allele frequency.


Ab initio quantum chemistry for protein structures.
Structural properties of over 55 small proteins have been determined using both density-based and wave-function-based electronic structure methods in order to assess the ability of ab initio "force fields" to retain the properties described by experimental structures measured with crystallography or nuclear magnetic resonance. The efficiency of the GPU-based quantum chemistry algorithms implemented in our TeraChem program enables us to carry out systematic optimization of ab initio protein structures, which we compare against experimental and molecular mechanics force field references. We show that the quality of the ab initio optimized structures, as judged by conventional protein health metrics, increases with increasing basis set size. On the other hand, there is little evidence for a significant improvement of predicted structures using density functional theory as compared to Hartree-Fock methods. Although occasional pathologies of minimal basis sets are observed, these are easily alleviated with even the smallest double-ζ basis sets.


Collaborative effects of chlorogenic acid and caffeine on lipid metabolism via the AMPKα-LXRα/SREBP-1c pathway in high-fat diet-induced obese mice.
To investigate the mechanism of the combined effects of chlorogenic acid (CGA) and caffeine on lipid metabolism in high-fat diet-induced obese mice, eighty female ICR mice were randomly divided into eight groups and fed with a high-fat diet with/without CGA and/or caffeine for 14 weeks. The combination of CGA and caffeine effectively decreased body weight gain, intraperitoneal adipose tissue weight, serum LDL-c, FFA, TC, TG, leptin, IL-6 concentrations, and hepatic TG and TC levels and increased the serum adiponectin level. The CGA and caffeine combination also promoted the phosphorylation of AMPKα, inhibited the expressions of transcriptional regulators (SREBP-1c and LXRα), and decreased the expressions of FAS and HMGR. Besides, the expressions of ACO, ATGL and HSL were increased by the CGA and caffeine combinations. The results indicated that the combination of CGA and caffeine had anti-obesity effects and regulated lipid metabolism in high-fat diet-induced obese mice via the AMPKα-LXRα/SREBP-1c signaling pathway. Thus, chronic CGA and caffeine intakes may be potent for preventing obesity.


Avoided deforestation as a greenhouse gas mitigation tool: economic issues.
Tropical deforestation is a significant contributor to accumulation of greenhouse gases (GHGs) in the atmosphere. GHG emissions from deforestation in the tropics were in the range of 1 to 2 Pg C yr(-1) for the 1990s, which is equivalent to as much as 25% of global anthropogenic GHG emissions. While there is growing interest in providing incentives to avoid deforestation and consequently reduce net carbon emissions, there is limited information available on the potential costs of these activities. This paper uses a global forestry and land use model to analyze the potential marginal costs of reducing net carbon emissions by avoiding deforestation in tropical countries. Our estimates suggest that about 0.1 Pg C yr(-1) of emissions reductions could be obtained over the next 30 to 50 yr for $5 per Mg C, and about 1.6 Pg C yr(-1) could be obtained over the same time frame for $100 per Mg C. In addition, the effects of carbon incentives on land use could be substantial. Relative to projected baseline conditions, we find that there would be around 3 million additional hectares (ha) of forestland in 2055 at $5 per Mg C and 422 million ha at $100 per Mg C. Estimates of reductions in area deforested, GHG mitigation potential, and annual land rental payments required are presented, all of which vary by region, carbon price paid, and time frame of mitigation.


Infants and the decision to provide ongoing child welfare services.
Infants are the most likely recipients of child welfare services; however, little is known about infants and families who come into contact with the child welfare system and factors that are associated with service provision. Investigations involving infants and their families present an unparalleled opportunity for the child welfare sector to enhance infants' safety and well-being through early identification, referral and intervention. Understanding how the child welfare system responds to the unique needs of infants and caregivers is critical to developing appropriate practice and policy responses within the child welfare sector and across other allied sectors. This study examines maltreatment-related investigations in Ontario involving children under the age of one to identify which factors are most influential to predicting service provision at the conclusion of a child welfare investigation. A secondary analysis of the fifth cycle of the Ontario Incidence Study of Reported Child Abuse and Neglect (OIS) for 2013 was conducted. The OIS is a cross-sectional child welfare study that is conducted every 5 years. The most influential factors that were associated with the decision to transfer a case to ongoing services were explored through a multivariate tree-classification technique, Chi square automatic interaction detection. There were an estimated 7915 maltreatment-related investigations involving infants in 2013. At least one caregiver risk factor was identified in approximately three-quarters (74%) of investigations involving infants. In the majority of investigations (57%), at least one referral for specialized services was provided. Primary caregiver with few social supports was the most highly significant predictor of the decision to provide ongoing child welfare services. Primary caregiver risk factors were predominant in this model. The analysis identified subgroups of investigations involving infants for which the likelihood of being transferred to ongoing services ranged from approximately 11-97%. Caregivers of infants are struggling with numerous challenges that can adversely compromise their ability to meet the unique developmental needs of their infant. The findings underscore the importance of community and social supports in decision-making.


Impact of centre volume, surgeon volume, surgeon experience and geographic location on reoperation after intramedullary nailing of tibial shaft fractures
Tibial shaft fractures are the most common long-bone injury, with a reported annual incidence of more than 75 000 in the United States. This study aimed to determine whether patients with tibial fractures managed with intramedullary nails experience a lower rate of reoperation if treated at higher-volume hospitals, or by higher-volume or more experienced surgeons. The Study to Prospectively Evaluate Reamed Intramedullary Nails in Patients with Tibial Fractures (SPRINT) was a multicentre randomized clinical trial comparing reamed and nonreamed intramedullary nailing on rates of reoperation to promote fracture union, treat infection or preserve the limb in patients with open and closed fractures of the tibial shaft. Using data from SPRINT, we quantified centre and surgeon volumes into quintiles. We performed analyses adjusted for type of fracture (open v. closed), type of injury (isolated v. multitrauma), gender and age for the primary outcome of reoperation using multivariable logistic regression. There were no significant differences in the odds of reoperation between high- and low-volume centres (p = 0.9). Overall, surgeon volume significantly affected the odds of reoperation (p = 0.03). The odds of reoperation among patients treated by moderate-volume surgeons were 50% less than those among patients treated by verylow-volume surgeons (odds ratio [OR] 0.50, 95% confidence interval [CI] 0.28–0.88), and the odds of reoperation among patients treated by high-volume surgeons were 47% less than those among patients treated by very-low-volume surgeons (OR 0.53, 95% CI 0.30–0.93). There appears to be no significant additional patient benefit in treatment by a higher-volume centre for intramedullary fixation of tibial shaft fractures. Additional research on the effects of surgical and clinical site volume in tibial shaft fracture management is needed to confirm this finding. The odds of reoperation were higher in patients treated by very-low-volume surgeons; this finding may be used to optimize the results of tibial shaft fracture management. Clinical trial registration: ClinicalTrials.gov, NCT00038129


The causal cognition of wrong doing: incest, intentionality, and morality.
The paper concerns the role of intentionality in reasoning about wrong doing. Anthropologists have claimed that, in certain non-Western societies, people ignore whether an act of wrong doing is committed intentionally or accidentally. To examine this proposition, we look at the case of Madagascar. We start by analyzing how Malagasy people respond to incest, and we find that in this case they do not seem to take intentionality into account: catastrophic consequences follow even if those who commit incest are not aware that they are related as kin; punishment befalls on innocent people; and the whole community is responsible for repairing the damage. However, by looking at how people reason about other types of wrong doing, we show that the role of intentionality is well understood, and that in fact this is so even in the case of incest. We therefore argue that, when people contemplate incest and its consequences, they simultaneously consider two quite different issues: the issue of intentionality and blame, and the much more troubling and dumbfounding issue of what society would be like if incest were to be permitted. This entails such a fundamental attack on kinship and on the very basis of society that issues of intentionality and blame become irrelevant. Using the insights we derive from this Malagasy case study, we re-examine the results of Haidt's psychological experiment on moral dumbfoundedness, which uses a story about incest between siblings as one of its test scenarios. We suggest that the dumbfoundedness that was documented among North American students may be explained by the same kind of complexity that we found in Madagascar. In light of this, we discuss the methodological limitations of experimental protocols, which are unable to grasp multiple levels of response. We also note the limitations of anthropological methods and the benefits of closer cross-disciplinary collaboration.


Prenatal nicotine exposure alters early pancreatic islet and adipose tissue development with consequences on the control of body weight and glucose metabolism later in life.
Despite medical advice, 20-30% of female smokers continue to smoke during pregnancy. Epidemiological studies have associated maternal smoking with increased risk of obesity and type-2 diabetes in the offspring. In the present study, we investigated the impact of prenatal nicotine exposure (3 mg/kg in Sprague Dawley rats via osmotic Alzet minipumps) on the early endocrine pancreas and adipose tissue development in rat pups before weaning. Body weight, fat deposition, food intake and food efficiency, cold tolerance, spontaneous physical activity, glucose utilization, and insulin sensitivity were also examined at adulthood. Prenatal nicotine exposure led to a decrease in endocrine pancreatic islet size and number at 7 d of life (postnatal d 7), which corroborates with a decrease in gene expression of specific transcription factors such as pancreatic and duodenal homeobox 1, Pax-6, Nkx6.1, and of hormones such as insulin and glucagon. The prenatal nicotine exposure also led to an increase in epididymal white adipose tissue weight at weaning (postnatal d 21), and marked hypertrophy of adipocytes, with increased gene expression of proadipogenic transcription factors such as CAAT-enhancer-binding protein-alpha, peroxisome proliferator activated receptor-gamma, and sterol regulatory element binding protein-1C. These early tissue alterations led to significant metabolic consequences, as shown by increased body weight and fat deposition, increased food efficiency on high-fat diet, cold intolerance, reduced physical activity, and glucose intolerance combined with insulin resistance observed at adulthood. These results prove a direct association between fetal nicotine exposure and offspring metabolic syndrome with early signs of dysregulations of adipose tissue and pancreatic development.


Influence of bone marrow stromal cell secreted molecules on pulpal and periodontal healing in replanted immature rat molars.
To investigate the effect of paracrine factors secreted from human bone marrow stromal cell (BMSC-CM) on pulpal and periodontal healing following immediate replantation of maxillary rat first molars. Fifty maxillary rat first molars were elevated and replanted after 2 min. The left teeth were replanted without treatment, whereas BMSC-CM was injected into the right socket prior to replantation. Twelve un-operated teeth served as reference teeth. The expression of vascular endothelial growth factor A, alkaline phosphatase, Runt-related transcription factor 2 and osteoclast stimulating factor 1 was studied by real-time reverse transcription polymerase chain reaction at day 3 and 14. The dentin thickness together with Laminin- and PGP 9.5-immunoreactivity were studied after 3, 14 and 90 days. Real-Time qRT-PCR data showed up-regulated expression of ALP mRNA in the socket specimens of conditioned medium treated replanted teeth after 3 days. No morphological differences were found for the expression of Laminin and PGP 9.5 between control and conditioned medium treated replanted teeth. At day 14, external cervical and surface root resorption was found in one BMSC-CM and one control tooth. At 90 days, all control replanted teeth had external cervical and surface root resorptions, whereas only one sample was seen among the conditioned medium treated teeth. At day 90, more extensive dentine formation with narrowing of the pulpal space was observed in the control compared with conditioned medium treated teeth. The present findings showed that BMSC-CM treatment reduced the number of replanted teeth with external root resorption and resulted in a significant reduction in new dentin formation.


Polymorphism in gene coding for ACE determines different development of myocardial fibrosis in rats.
In humans, the effect of angiotensin-converting enzyme (ACE) gene polymorphisms in cardiovascular disease is still controversial. In the rat, a microsatellite marker in the ACE gene allows differentiation of the ACE gene polymorphism among strains with different ACE levels. We tested the hypothesis that this ACE gene polymorphism determines the extent of cardiac fibrosis induced by isoproterenol (Iso) in the rat. We used a male F(2) generation (homozygous LL and BB ACE genotypes determined by polymerase chain reaction) derived from two rat strains [Brown-Norway (BB) and Lewis (LL)] that differ with respect to their plasma ACE activities. For induction of left ventricular (LV) hypertrophy (LVH) and cardiac fibrosis, rats were infused with Iso (5 mg x kg(-1) x day(-1)) or saline (control) for 10 days and euthanized at day 1 after the last injection. The interstitial collagen volumetric fraction (ICVF), collagen I, and fibronectin content, but not collagen III content, were significantly higher in the homozygous BB rats than in homozygous LL rats. Differences in metalloprotease (MMP)-9, but not in MMP-2 activities as well as in cardiac cell proliferation, were also detected between LL and BB rats treated with Iso. LV ACE activity was higher in BB rats than LL rats and correlated with ICVF (r = 0.61, P < 0.002). No changes were observed in plasma ACE activities, ANG II plasma or LV levels, plasma renin activity, and ACE and ANG II type 1 receptor (AT1R) mRNA levels in the LV of rats with the two different ACE polymorphisms. Iso induced a similar degree of LVH [assessed by an increase in LV weight 100 per body weight, LV-to-right ventricle (RV) ratio, and LV protein content] in LL and BB rats. We concluded that rats in the F(2) generation with high plasma ACE activity developed more fibrosis but to a similar degree of LVH compared with rats with low plasma ACE activity.


Prepulse inhibition deficits and perseverative motor patterns in dopamine transporter knock-out mice: differential effects of D1 and D2 receptor antagonists.
Dopamine is known to regulate several behavioral phenomena, including sensorimotor gating and aspects of motor activity. The roles of dopamine D1 and D2 receptors in these behaviors have been documented in the rat literature, but few reports exist on their role in mice. We used dopamine transporter (DAT) (-/-) mice to examine the behavioral consequences of a chronically hyperdopaminergic state, challenging them with the preferential dopamine D2 receptor antagonist raclopride and D1 receptor antagonist SCH23390. At baseline, DAT (-/-) mice exhibited deficient sensorimotor gating as measured by prepulse inhibition (PPI) of the startle response, exhibited nonfocal preservative patterns of locomotion, and were hyperactive in a novel environment. Pretreatment with raclopride significantly increased PPI in the DAT (-/-) mice, whereas SCH23390 had no significant effect. Blockade of D2 receptors did not affect the predominantly straight patterns of motor behavior produced by the DAT (-/-) mice, but antagonism of D1 receptors significantly attenuated the preservative patterns, producing more of a meandering behavior seen in the DAT (+/+) control mice. Both D1 and D2 receptor antagonists decreased the hyperactivity seen in the DAT (-/-) mice. These findings support the role of the D2, but not the D1, receptor in the modulation of PPI in mice. Furthermore, D1 receptor activation appears to be the critical substrate for the expression of preservative patterns of motor behavior, whereas both D1 and D2 receptors appear to regulate the amount of motor activity.


Generation of a baculovirus recombinant prostate-specific membrane antigen and its use in the development of a novel protein biochip quantitative immunoassay.
Prostate-specific membrane antigen (PSMA) is a 100-kDa transmembrane glycoprotein identified by the monoclonal antibody 7E11-C5.3 from the human prostate tumor cell line LNCaP. Because of its significant upregulation in androgen refractory and metastatic prostate cancers, PSMA may be a useful prognostic biomarker and a target for developing novel therapeutic strategies. However, the lack of abundant pure PSMA protein and the low efficacy in immunoaffinity isolation from LNCaP cells have hampered the development of clinical assays. In order to obtain a renewable and reliable source of pure antigen, we used the baculovirus/insect cell system to express and purify a recombinant PSMA. A recombinant baculovirus containing a 6x histidine-tagged PSMA gene was generated, from which rPSMA was expressed and purified using cobalt-chelating affinity chromatography. The purity and correct molecular size of rPSMA were demonstrated by gel electrophoresis and mass spectrometry. Glycosidase digestions showed that the oligosaccharides on rPSMA are primarily N-linked high-mannose type. Although the glycosylation is different from the native PSMA, it did not affect the immunoreactivity of rPSMA to antibodies specific for either the intra- or the extracellular domains of PSMA. Finally, the purified rPSMA was successfully used to develop a quantitative PSMA immunoassay using the novel ProteinChip surface-enhanced laser desorption/ionization mass spectrometry technology.


Alpha-fetoprotein, des-gamma carboxyprothrombin, and lectin-bound alpha-fetoprotein in early hepatocellular carcinoma.
Alpha-fetoprotein (AFP) is widely used as a surveillance test for hepatocellular carcinoma (HCC) among patients with cirrhosis. Des-gamma carboxy-prothrombin (DCP) and lectin-bound AFP (AFP-L3%) are potential surveillance tests for HCC. The aims of this study were to determine performance of DCP and AFP-L3% for the diagnosis of early HCC; whether they complement AFP; and what factors affect DCP, AFP-L3%, or AFP levels. We conducted a large phase 2 biomarker case-control study in 7 academic medical centers in the United States. Controls were patients with compensated cirrhosis and cases were patients with HCC. AFP, DCP, and AFP-L3% levels were measured blinded to clinical data in a central reference laboratory. A total of 836 patients were enrolled: 417 (50%) were cirrhosis controls and 419 (50%) were HCC cases, of which 208 (49.6%) had early stage HCC (n = 77 very early, n = 131 early). AFP had the best area under the receiver operating characteristic curve (0.80, 95% confidence interval [CI]: 0.77-0.84), followed by DCP (0.72, 95% CI: 0.68-0.77) and AFP-L3% (0.66, 95% CI: 0.62-0.70) for early stage HCC. The optimal AFP cutoff value was 10.9 ng/mL leading to a sensitivity of 66%. When only those with very early HCC were evaluated, the area under the receiver operating characteristic curve for AFP was 0.78 (95% CI: 0.72-0.85) leading to a sensitivity of 65% at the same cutoff. AFP was more sensitive than DCP and AFP-L3% for the diagnosis of early and very early stage HCC at a new cutoff of 10.9 ng/mL.


Characterization of the primary sonic muscles in Carapus acus (Carapidae): a multidisciplinary approach.
Sound production in carapid fishes results from the action of extrinsic muscles that insert into the swim bladder. Biochemical, histochemical and morphological techniques were used to examine the sonic muscles and compare them with epaxial muscles in Carapus acus. Sonic fibres are thicker than red and thinner than white epaxial fibres, and sonic fibres and myofibrils exhibit an unusual helicoidal organization: the myofibrils of the centre are in a straight line whereas they are more and more twisted towards the periphery. Sonic muscles have both features of red (numerous mitochondria, high glycogen content) and white (alkali-stable ATPase) fibres. They differ also in the isoforms of the light chain (LC3) and heavy chain (HC), in having T tubules at both the Z-line and the A-I junction and in a unique parvalbumin isoform (PAI) that may aid relaxation. All these features lead to the expression of two assumptions about sound generation: the sonic muscle should be able to perform fast and powerful contractions that provoke the forward movement of the forepart of the swim bladder and the stretching and "flapping" of the swim bladder fenestra; the helicoidal organization allows progressive drawing of the swim bladder fenestra which emits a sound when rapidly released in a spring-like manner.


Manipulation or mobilisation for neck pain.
Manipulation and mobilisation are often used, either alone or combined with other treatment approaches, to treat neck pain. To assess if manipulation or mobilisation improves pain, function/disability, patient satisfaction, quality of life, and global perceived effect in adults with acute/subacute/chronic neck pain with or without cervicogenic headache or radicular findings. CENTRAL (The Cochrane Library 2009, issue 3) and MEDLINE, EMBASE, Manual Alternative and Natural Therapy, CINAHL, and Index to Chiropractic Literature were updated to July 2009. Randomised controlled trials on manipulation or mobilisation. Two review authors independently selected studies, abstracted data, and assessed risk of bias. Pooled relative risk and standardised mean differences (SMD) were calculated. We included 27 trials (1522 participants).Cervical Manipulation for subacute/chronic neck pain : Moderate quality evidence suggested manipulation and mobilisation produced similar effects on pain, function and patient satisfaction at intermediate-term follow-up. Low quality evidence showed manipulation alone compared to a control may provide short- term relief following one to four sessions (SMD pooled -0.90 (95%CI: -1.78 to -0.02)) and that nine or 12 sessions were superior to three for pain and disability in cervicogenic headache. Optimal technique and dose need to be determined.Thoracic Manipulation for acute/chronic neck pain : Low quality evidence supported thoracic manipulation as an additional therapy for pain reduction (NNT 7; 46.6% treatment advantage) and increased function (NNT 5; 40.6% treatment advantage) in acute pain and favoured a single session of thoracic manipulation for immediate pain reduction compared to placebo for chronic neck pain (NNT 5, 29% treatment advantage).Mobilisation for subacute/chronic neck pain: In addition to the evidence noted above, low quality evidence for subacute and chronic neck pain indicated that 1) a combination of Maitland mobilisation techniques was similar to acupuncture for immediate pain relief and increased function; 2) there was no difference between mobilisation and acupuncture as additional treatments for immediate pain relief and improved function; and 3) neural dynamic mobilisations may produce clinically important reduction of pain immediately post-treatment. Certain mobilisation techniques were superior. Cervical manipulation and mobilisation produced similar changes. Either may provide immediate- or short-term change; no long-term data are available. Thoracic manipulation may improve pain and function. Optimal techniques and dose are unresolved. Further research is very likely to have an important impact on our confidence in the estimate of effect and is likely to change the estimate.


Pancreatic endocrine tumors are a rare manifestation of the neurofibromatosis type 1 phenotype: molecular analysis of a malignant insulinoma in a NF-1 patient.
The tumorigenesis of sporadic endocrine tumors is still not fully understood. It is well known that patients with von Recklinghausen syndrome (NF-1) (OMIM 162200) carrying NF1 germline mutations are predisposed to endocrine tumors including pheochromocytomas and duodenal somatostatinomas. It is unclear, however, whether the rarely reported occurrence of pancreatic insulinomas in NF-1 patients represents a coincidental finding or whether insulinomas are a rare manifestation of the NF-1 syndrome. To determine the potential association between the NF-1 syndrome and pancreatic endocrine tumors, we analyzed a NF-1 patient with a well-differentiated pancreatic endocrine carcinoma for NF1 mutation, allelic loss of the NF1 gene and its expression in peripheral blood and tumor cells. The germline mutation c. 499 del TGTT known in the family was confirmed by polymerase chain reaction (PCR) and direct sequencing of exon 4 in DNA extracted from peripheral blood. Loss of heterozygosity (LOH) analysis of the NF1 gene was carried out using 3 intragenic microsatellite markers on 17q11.2. RNA expression was examined by reverse transcription and a consecutive PCR spanning intron 3 of the NF1 gene including the mutated site in exon 4. Immunohistochemistry was used to analyze NF-1 protein expression. Mutation analysis of peripheral blood leukocytes confirmed the 4 base pair deletion in exon 4 starting at codon 167 (499 del TGTT). LOH analysis of tumor tissue revealed retention of both NF1 alleles. While reverse transcriptase-PCR of peripheral blood showed bi-allelic expression of both the wild-type NF1 and the mutated form, reverse transcriptase-PCR of tumor extracts demonstrated expression of the mutated but not the wild-type NF1 allele. Additionally, neurofibromin, the NF1 gene product, was absent in the tumor tissue of the NF-1 patient. These results show that the wild-type NF1 transcrips and protein are reduced, in the reported insulinoma, supposedly by epigenetic mechanisms. This provides strong evidence that there is a relationship between von Recklinghausen disease and the patient's insulinoma. In this line, insulinomas may be viewed as a rare manifestation of the NF-1 syndrome. Furthermore, the NF1 gene must be considered as a candidate tumor suppressor gene for sporadic insulinomas and probably other pancreatic endocrine tumors.


Intention to quit and the role of dark personality and perceived organizational support: A moderation and mediation model.
This study investigated the role of individual differences (dark personality) and situational factors (perceived organisational support) in explaining intention to quit. Four hundred and fifty-one (50 of which females) ambulance personnel completed three questionnaires (Hogan Development Survey; Perceived Organisational Support Survey; and a single item Intention to Quit measure) as a part of a selection and development assessment. Employees high on Excitable, Sceptical, and Mischievous, but low on Colourful were found to have greater intentions to quit. Additionally, employees high on Excitable, Sceptical, Reserved, and Leisurely, but low on Dutiful and Diligent had lower perceptions of organisational support. Structural Equation Modelling revealed that perceived organisational support plays both a mediating and moderating role on dark personality and intention to quit. Theoretical implications of personality's role in perceived organisational support and intention to quit are discussed.


NAFLD in children: a prospective clinical-pathological study and effect of lifestyle advice.
Nonalcoholic fatty liver disease (NAFLD), a common cause of chronic liver disease in adults, is incompletely characterized in children. We conducted a prospective study to better characterize the clinical presentation of NAFLD in children and to determine the effect of lifestyle advice in the management of pediatric NAFLD. From June 2001 to April 2003, 84 children (age 3-18.8 yr) who had elevated aminotransferases and the diagnosis of NAFLD confirmed via liver biopsy underwent a 2-hour oral glucose tolerance test and a 12-month program of lifestyle advice consisting of diet and physical exercise. Thirty-four (40.5%) patients were obese (body mass index [BMI] >97th percentile), and 43 (51.2%) were overweight (BMI 85th-97th percentile). Ten (12%) had abnormal glucose tolerance; 10 (12%) had elevated triglycerides, cholesterol, or both; and all had normal blood pressure. Most children (67/84, 80%) were insulin-resistant, including the 7 children with normal BMI (<85th percentile). Increased liver fibrosis was present in 49 (58.1%) patients and was independently associated with obesity (OR 2.7, 95% CI 1.2-6.2) and age (1-year increase; OR 1.2, 95% CI 1.04-1.5). A 12-month program with diet and physical exercise resulted in a significant decrease in BMI, and levels of fasting glucose, insulin, lipids, and liver enzymes, as well as liver echogenicity on ultrasonography. In conclusion, children with NAFLD are almost always insulin-resistant regardless of BMI. Obesity and older age are independently associated with increased liver fibrosis. A simple lifestyle advice program significantly improves insulin resistance, and the liver disease in pediatric NAFLD.


The transformation of American midwifery: 1975 to 1988.
The use of midwives is a natural solution to the problem of improving access to skilled perinatal services while lowering costs. The number of midwife-attended births has grown from 0.9% of all births in 1975 to 3.4% of all births in 1988. The purpose of the study was to determine how mothers served by midwives and the settings in which they are served have changed in that period. The analysis is based on birth certificate data from 1975 to 1988 from the Natality, Marriage and Divorce Statistics Branch of the National Center for Health Statistics, Centers for Disease Control. Almost all of the growth (93.2%) in midwife-attended births from 1975 to 1988 was in hospitals; 87.3% of all births attended by midwives occurred in hospitals. Pronounced differences exist between mothers served by midwives in and outside of hospitals, and there are strong regional patterns in midwife attendance at birth. Given the positive outcomes associated with midwifery practice, further research into the content of midwifery care is recommended.


Measuring the effect of intimate partner violence on health-related quality of life: a qualitative focus group study.
Health related quality of life (HRQOL) can be measured by a wide range of instruments, many of which have been designed for specific conditions or uses. "Preference-based" measures assess the value individuals place on health, and are included in economic evaluations of treatments and interventions (such as cost effectiveness analysis). As economic evaluation becomes more common, it is important to assess the applicability of preference-based health related quality of life (HRQOL) measures to public health issues. This study investigated the usefulness of such instruments in the context of intimate partner violence (IPV), a public health concern that that can seriously affect quality of life. The study consisted of focus groups with abused women to determine the aspects of life affected by IPV, and an analysis of existing HRQOL measures. Eight focus groups (n = 40) were conducted in which participants discussed the domains of health affected by IPV. Results were content analyzed and compared with the domains of health included in four commonly-used, preference-based HRQOL measures. The average focus group participant was 43 years old, unemployed, African American, with 3 children. Domains of health reported to be affected by IPV included physical functioning, emotional and psychological functioning, social functioning and children's functioning. Psychological health was the most severely affected domain. The Short Form 36, the Health Utilities Index, the EuroQol 5D, and the Quality of Well-being Scale were found to vary in the degree to which they include domains of health important in IPV. Psychological health is included to a limited extent, and the spill-over effect of a condition on other family members, including children, is not included at all. Emotional and psychological health plays an important role in the overall HRQOL of abused women but is relatively underemphasized in preference-based HRQOL measures. This may lead to an underestimation of the impact of partner violence on HRQOL when using these measures and in economic evaluations that rely thereon. Holistic measurement approaches or expanded measures that capture the far-reaching effects of IPV on HRQOL may be needed to accurately measure the effect of this condition on women's health.


Truncated RanGAP encoded by the Segregation Distorter locus of Drosophila.
Segregation Distorter (SD) in Drosophila melanogaster is a naturally occurring meiotic drive system in which the SD chromosome is transmitted from SD/SD+ males in vast excess over its homolog owing to the induced dysfunction of SD+-bearing spermatids. The Sd locus is the key distorting gene responsible for this phenotype. A genomic fragment from the Sd region conferred full distorting activity when introduced into the appropriate genetic background by germline transformation. The only functional product encoded by this fragment is a truncated version of the RanGAP nuclear transport protein. These results demonstrate that this mutant RanGAP is the functional Sd product.


Novel biological treatments for systemic lupus erythematosus: current and future modalities.
Current treatments for systemic lupus erythematosus (SLE) are effective in reducing morbidity and mortality but are not specific and have severe adverse effects. Based on understanding of the different dysregulated immunological pathways involved in SLE pathogenesis, specific targeted therapies were developed. This review presents the current and the near-future novel biological immune targeted treatments, such as B cell-targeted therapy, cytokine blockade, peptide-based treatments and other novel treatments for SLE.


Lec3 Chinese hamster ovary mutants lack UDP-N-acetylglucosamine 2-epimerase activity because of mutations in the epimerase domain of the Gne gene.
Lec3 Chinese hamster ovary (CHO) cell glycosylation mutants have a defect in sialic acid biosynthesis that is shown here to be reflected most sensitively in reduced polysialic acid (PSA) on neural cell adhesion molecules. To identify the genetic origin of the phenotype, genes encoding different factors required for sialic acid biosynthesis were transfected into Lec3 cells. Only a Gne cDNA encoding UDP-GlcNAc 2-epimerase:ManNAc kinase rescued PSA synthesis. In an in vitro UDP-GlcNAc 2-epimerase assay, Lec3 cells had no detectable UDP-GlcNAc 2-epimerase activity, and Lec3 cells grown in serum-free medium were essentially devoid of sialic acid on glycoproteins. The Lec3 phenotype was rescued by exogenously added N-acetylmannosamine or mannosamine but not by the same concentrations of N-acetylglucosamine, glucosamine, glucose, or mannose. Sequencing of CHO Gne cDNAs identified a nonsense (E35stop) and a missense (G135E) mutation, respectively, in two independent Lec3 mutants. The G135E Lec3 mutant transfected with a rat Gne cDNA had restored in vitro UDP-GlcNAc 2-epimerase activity and cell surface PSA expression. Both Lec3 mutants were similarly rescued with a CHO Gne cDNA and with CHO Gne encoding the known kinase-deficient D413K mutation. However, cDNAs encoding the known epimerase-deficient mutation H132A or the new Lec3 G135E Gne mutation did not rescue the Lec3 phenotype. The G135E Gne missense mutation is a novel mechanism for inactivating UDP-GlcNAc 2-epimerase activity. Lec3 mutants with no UDP-GlcNAc 2-epimerase activity represent sensitive hosts for characterizing disease-causing mutations in the human GNE gene that give rise to sialuria, hereditary inclusion body myopathy, and Nonaka myopathy.


A prospective randomized trial of ultrasound- vs landmark-guided central venous access in the pediatric population.
The purpose of this prospective randomized study was to compare landmark- to ultrasound-guided central venous access when performed by pediatric surgeons. The American College of Surgeons advocates for use of ultrasound in central venous catheter placement; however, this is not universally embraced by pediatric surgeons. Complication risk correlates positively with number of venous cannulation attempts. With IRB approval, a randomized prospective study of children under 18 years of age undergoing tunneled central venous catheter placement was performed. Patient accrual was based on power analysis. Exclusion criteria included known nonpatency of a central vein or coagulopathy. After randomization, the patients were assigned to either ultrasound-guided internal jugular vein access or landmark-guided subclavian/internal jugular vein access. The primary outcomes measure was number of attempts at venous cannulation. Secondary outcomes measures included: access times, number of arterial punctures, and other complications. Continuous variables were compared using 2-tailed Student's t-test. Discrete variables were analyzed with chi-square. Significance was defined as p < 0.05. There were 150 patients enrolled between April 2008 and September 2011. There was no difference when comparing demographic data. Success at first attempt was achieved in 65% of patients in the ultrasound group vs 45% in the landmark group (p = 0.021). Success within 3 attempts was achieved in 95% of ultrasound group vs 74% of landmark group (p = 0.0001). Ultrasound reduced the number of cannulation attempts necessary for venous access. This indicates a potential to reduce complications when ultrasound is used by pediatric surgeons.


Hyperbaric oxygen treatment augments the efficacy of a losartan regime in an experimental nephrotic syndrome model.
Proteinuria is associated with oxidant stress and inflammation. Hyperbaric oxygen (HBO) treatment has anti-inflammatory and anti-oxidant effects. The aim of the study was to investigate the benefits of HBO treatment on an experimental nephrotic syndrome model. 50 male Sprague-Dawley rats weighing 255 +/- 39 g were housed. Forty rats were injected 6 mg/kg adriamycin into tail veins under anesthesia to induce nephrosis, while 10 rats were spared as sham control. After the stabilization of proteinuria at the sixth week, the rats were treated for 6 weeks by losartan (n = 10, 30 mg/kg/day), HBO (n = 10, 2.8 atmosphere absolute, 90 min/day), HBO + losartan (n = 10) and vehicle (n = 10). Protein carbonyl (PCO), superoxide dismutase (SOD) and glutathione peroxidase (GPx) were analyzed from tissue specimens. Biochemical markers were studied from venous samples and 24-hour urine was collected for proteinuria. The surviving animals at 12 weeks (vehicle group (n = 6), HBO (n = 6), losartan (n = 8), HBO + losartan (n = 10) were sacrificed. Glomerular sclerosis, tubulointerstitial and blood vessel changes were determined by semiquantitative scoring. The PCO levels increased (p < 0.001), and the GPx and SOD levels decreased (p < 0.001 for both) in the nephrotic rats. In losartan and HBO groups GPx levels increased (p = 0.001, p = 0.002 respectively), but PCO and SOD levels did not change. The combination of HBO with losartan significantly increased the GPx and SOD levels (p = 0.001 for both) and decreased PCO levels (p = 0.005). HBO but not losartan significantly reduced proteinuria (p < 0.001). The combination of HBO and losartan reduced proteinuria better than the single losartan regime (p < 0.001). The effect of the combination was also noticed on the histological examination of the kidneys. The activities, appetites, weight gains, and improvement of edema were better in the HBO combined with losartan regime. These results indicate that the addition of HBO therapy to a conventional regime, angiotensin receptor blockers, has significant benefits in the management of proteinuria. Future clinical studies are needed to elucidate the role of HBO and other antioxidant strategies in the treatment of proteinuria.


Screening mosquito house entry points as a potential method for integrated control of endophagic filariasis, arbovirus and malaria vectors.
Partial mosquito-proofing of houses with screens and ceilings has the potential to reduce indoor densities of malaria mosquitoes. We wish to measure whether it will also reduce indoor densities of vectors of neglected tropical diseases. The main house entry points preferred by anopheline and culicine vectors were determined through controlled experiments using specially designed experimental huts and village houses in Lupiro village, southern Tanzania. The benefit of screening different entry points (eaves, windows and doors) using PVC-coated fibre glass netting material in terms of reduced indoor densities of mosquitoes was evaluated compared to the control. 23,027 mosquitoes were caught with CDC light traps; 77.9% (17,929) were Anopheles gambiae sensu lato, of which 66.2% were An. arabiensis and 33.8% An. gambiae sensu stricto. The remainder comprised 0.2% (50) An. funestus, 10.2% (2359) Culex spp. and 11.6% (2664) Mansonia spp. Screening eaves reduced densities of Anopheles gambiae s. l. (Relative ratio (RR) = 0.91; 95% CI = 0.84, 0.98; P = 0.01); Mansonia africana (RR = 0.43; 95% CI = 0.26, 0.76; P<0.001) and Mansonia uniformis (RR = 0.37; 95% CI = 0.25, 0.56; P<0.001) but not Culex quinquefasciatus, Cx. univittatus or Cx. theileri. Numbers of these species were reduced by screening windows and doors but this was not significant. This study confirms that across Africa, screening eaves protects households against important mosquito vectors of filariasis, Rift Valley Fever and O'Nyong nyong as well as malaria. While full house screening is required to exclude Culex species mosquitoes, screening of eaves alone or fitting ceilings has considerable potential for integrated control of other vectors of filariasis, arbovirus and malaria.


Excessive fructose intake induces the features of metabolic syndrome in healthy adult men: role of uric acid in the hypertensive response.
Excessive fructose intake causes metabolic syndrome in animals and can be partially prevented by lowering the uric acid level. We tested the hypothesis that fructose might induce features of metabolic syndrome in adult men and whether this is protected by allopurinol. A randomized, controlled trial of 74 adult men who were administered 200 g fructose daily for 2 weeks with or without allopurinol. Primary measures included changes in ambulatory blood pressure (BP), fasting lipids, glucose and insulin, homeostatic model assessment (HOMA) index, body mass index and criteria for metabolic syndrome. The ingestion of fructose resulted in an increase in ambulatory BP (7+/-2 and 5+/-2 mm Hg for systolic (SBP) and diastolic BP (DBP), P<0.004 and P<0.007, respectively). Mean fasting triglycerides increased by 0.62+/-0.23 mmol l(-1) (55+/-20 mg per 100 ml), whereas high-density lipoprotein cholesterol decreased by 0.06+/-0.02 mmol l(-1) (2.5+/-0.7 mg per 100 ml), P<0.002 and P<0.001, respectively. Fasting insulin and HOMA indices increased significantly, whereas plasma glucose level did not change. All liver function tests showed an increase in values. The metabolic syndrome increased by 25-33% depending on the criteria. Allopurinol lowered the serum uric acid level (P<0.0001) and prevented the increase in 24-h ambulatory DBP and daytime SBP and DBP. Allopurinol treatment did not reduce HOMA or fasting plasma triglyceride levels, but lowered low-density lipoprotein cholesterol relative to control (P<0.02) and also prevented the increase in newly diagnosed metabolic syndrome (0-2%, P=0.009). High doses of fructose raise the BP and cause the features of metabolic syndrome. Lowering the uric acid level prevents the increase in mean arterial blood pressure. Excessive intake of fructose may have a role in the current epidemics of obesity and diabetes.


Rare expression of KIT and absence of KIT mutations in high grade renal cell carcinoma.
RCCs with sarcomatoid differentiation have been reported to frequently express KIT protein, suggesting that imatinib mesylate (STI-571 or Gleevectrade mark) may be effective treatment for these aggressive tumors. We determined the frequency of KIT expression and mutation in a large series of high grade RCCs. We identified 194 patients who underwent nephrectomy for unilateral, sporadic nuclear grade 4 RCC between 1970 and 2002, including 123 with sarcomatoid differentiation. Sections from representative paraffin embedded tissue blocks were immunostained in an autostainer using ethylenediaminetetraacetic acid antigen retrieval, a polyclonal KIT antibody and the avidin-biotin peroxidase complex method. Mutational analysis was performed in all immunopositive and select negative cases by polymerase chain reaction amplification of KIT exons 9, 11, 13 and 17. Only 7 tumors (3.6%) showed KIT expression, including 5 of the 123 (4.1%) with sarcomatoid differentiation. Four of the 7 tumors showed focal staining only. No mutations were identified in the 7 positive cases or in 8 randomly selected negative samples. Death from RCC occurred in all 7 patients with KIT positive tumors at a median of 0.6 years (range 0.3 to 2.3) and in 139 of 187 with KIT negative tumors at a median of 0.8 years (range 0 to 10.2). KIT expression was identified in less than 5% of high grade RCCs with or without sarcomatoid differentiation but none of the tumors showed KIT mutations. These findings indicate that imatinib therapy is unlikely to be effective in patients with high grade RCC.


Geographical access to care at birth in Ghana: a barrier to safe motherhood.
Appropriate facility-based care at birth is a key determinant of safe motherhood but geographical access remains poor in many high burden regions. Despite its importance, geographical access is rarely audited systematically, preventing integration in national-level maternal health system assessment and planning. In this study, we develop a uniquely detailed set of spatially-linked data and a calibrated geospatial model to undertake a national-scale audit of geographical access to maternity care at birth in Ghana, a high-burden country typical of many in sub-Saharan Africa. We assembled detailed spatial data on the population, health facilities, and landscape features influencing journeys. These were used in a geospatial model to estimate journey-time for all women of childbearing age (WoCBA) to their nearest health facility offering differing levels of care at birth, taking into account different transport types and availability. We calibrated the model using data on actual journeys made by women seeking care. We found that a third of women (34%) in Ghana live beyond the clinically significant two-hour threshold from facilities likely to offer emergency obstetric and neonatal care (EmONC) classed at the 'partial' standard or better. Nearly half (45%) live that distance or further from 'comprehensive' EmONC facilities, offering life-saving blood transfusion and surgery. In the most remote regions these figures rose to 63% and 81%, respectively. Poor levels of access were found in many regions that meet international targets based on facilities-per-capita ratios. Detailed data assembly combined with geospatial modelling can provide nation-wide audits of geographical access to care at birth to support systemic maternal health planning, human resource deployment, and strategic targeting. Current international benchmarks of maternal health care provision are inadequate for these purposes because they fail to take account of the location and accessibility of services relative to the women they serve.


[Effects of D-limonene on leukemia cells HL-60 and K562 in vitro].
To investigate the effects of D-limonene (D-L) on the cell growth and apoptosis in HL-60, K562 cells and to elucidate its mechanism, the influence of D-L on proliferation of HL-60 and K562 cells was determined by propidium iodide assay, the expression levels of mutant p53, bcl-2, bax gene were detected by cell morphological analysis, flow cytometry and immunohistochemistry staining, the D-L-inducing HL-60 and K562 cell apoptosis in vitro was observed systematically. The results showed that D-L inhibited HL-60 and K562 cell growth in a dose- and time-dependent manner with the IC50 of 0.75 mmol/L similarly, D-L induced apoptosis of HL-60 and K562 cells, and expression of bcl-2 gene was down regulated by D-L in a concentration-dependent manner in HL-60 cells. The bcl-2, mutant type of p53 genes were down regulated while bax gene was up regulated by D-L in a concentration-dependent manner in K562 cells. It is concluded that D-L can inhibit proliferation and induce apoptosis of HL-60 and K562 cells. The bcl-2, mutant type of p53 and bax may be involved in the gene regulation of D-L-induced apoptosis.


A triplet spike-timing-dependent plasticity model generalizes the Bienenstock-Cooper-Munro rule to higher-order spatiotemporal correlations.
Synaptic strength depresses for low and potentiates for high activation of the postsynaptic neuron. This feature is a key property of the Bienenstock-Cooper-Munro (BCM) synaptic learning rule, which has been shown to maximize the selectivity of the postsynaptic neuron, and thereby offers a possible explanation for experience-dependent cortical plasticity such as orientation selectivity. However, the BCM framework is rate-based and a significant amount of recent work has shown that synaptic plasticity also depends on the precise timing of presynaptic and postsynaptic spikes. Here we consider a triplet model of spike-timing-dependent plasticity (STDP) that depends on the interactions of three precisely timed spikes. Triplet STDP has been shown to describe plasticity experiments that the classical STDP rule, based on pairs of spikes, has failed to capture. In the case of rate-based patterns, we show a tight correspondence between the triplet STDP rule and the BCM rule. We analytically demonstrate the selectivity property of the triplet STDP rule for orthogonal inputs and perform numerical simulations for nonorthogonal inputs. Moreover, in contrast to BCM, we show that triplet STDP can also induce selectivity for input patterns consisting of higher-order spatiotemporal correlations, which exist in natural stimuli and have been measured in the brain. We show that this sensitivity to higher-order correlations can be used to develop direction and speed selectivity.


Partial ovary development is widespread in honey bees and comparable to other eusocial bees and wasps.
Honey bee workers have few opportunities for direct reproduction because their ovary development is chemically suppressed by queens and worker-laid eggs are destroyed by workers. While workers with fully developed ovaries are rare in honey bee colonies, we show that partial ovary development is common. Across nine studies, an average of 6% to 43% of workers had partially developed ovaries in queenright colonies with naturally mated queens. This shift by workers toward potential future reproduction is linked to lower productivity, which suggests that even small investments in reproductive physiology by selfish workers reduce cooperation below a theoretical maximum. Furthermore, comparisons across 26 species of bees and wasps revealed that the level of partial ovary development in honey bees is similar to that of other eusocial Hymenoptera where there is reproductive conflict among colony members. Natural variation in the extent of partial ovary development in honey bee colonies calls for an exploration of the genetic and ecological factors that modulate shifts in cooperation within animal societies.


First Report of Fusarium oxysporum Causing Wilt on Jade Plant (Crassula ovata) in Italy.
During summer 2010, symptoms of a wilt disease were observed in a commercial farm in northern Italy on Crassula ovata (jade plant). First symptoms consisted of chlorosis and premature drop of still turgid leaves. As the disease progressed, leaves turned yellow and wilted before dropping off and the stem wilted, bent, and eventually rotted starting from the base. In some cases, the stem broke or the basal portion of the leaf rotted. Brown discolorations were observed in the vascular system. Of 10,000 plants, 65% (cv. Mini) and 5% of 600 plants (cv. Magical Tree) were affected. Premature dropping of leaves was more frequent on cv. Magical Tree. Using the Komada's Fusarium-selective medium, a fungus was consistently and readily isolated from symptomatic vascular tissues of plants belonging to both cultivars. Isolates obtained from both cultivars were purified, subcultured on potato dextrose agar (PDA), and single-spore cultures were obtained. On PDA, both isolates produced pale violet, abundant, aerial mycelium, felted in old cultures, with purple pigments in the agar. The isolates were grown on Spezieller Nährstoffarmer agar for characterization of macroconidia and microconidia (1). Both isolates produced sparse, 3 to 5 septate, nearly straight macroconidia measuring 30 to 56 × 3 to 5 (average 40 × 4) μm with a short apical cell and a foot-shaped basal cell. Sporodochia were not observed. Unicellular, oval-elliptical microconidia measuring 5 to 13 × 3 to 4 (average 8 × 3) μm were produced on short monophialides. Chlamydospores were abundant, single and sometime in pairs, terminal and intercalary, rough walled, and measured 6 to 9 μm. Such characteristics are typical of Fusarium oxysporum (3). The ITS region (internal transcribed spacer) of rDNA was amplified with primers ITS1/ITS4 (4) and sequenced. BLASTn analysis of an isolate from C. ovata cv. Mini (515 bp, Accession No. HQ682196) and C. ovata cv. Magical Tree (509 bp, Accession No. HQ682197) showed an E-value of 0.0 with F. oxysporum. For these sequences, pairwise alignment of EMBOSS (E.B.I. - The European Bioinformatics Institute) revealed identity and similarity of 99.0%. To confirm pathogenicity, tests were conducted on 5-month-old plants of cvs. Mini and Magical Tree. Plants (three per treatment) were inoculated by dipping roots in a 1 × 106 CFU/ml conidial suspension of the two isolates of F. oxysporum prepared from 10-day-old cultures grown on casein liquid medium (2), shaken (90 rpm) for 10 days at 24°C ± 1 (12-h fluorescent light, 12-h dark). Inoculated plants were transplanted into pots filled with steamed mix (sphagnum peat/perlite/pine bark/clay; 50:20:20:10) and maintained in a plant growth chamber at 25 ± 1°C under a regimen of 12 h per day of fluorescent light. Inoculated plants belonging to both cultivars showed typical first symptoms of Fusarium wilt after 13 days. In the following days, leaves dropped, stems wilted, and plants died. Noninoculated plants remained healthy. F. oxysporum was reisolated from inoculated plants. The pathogenicity test was conducted twice. This is, to our knowledge, the first report of F. oxysporum on C. ovata in Italy or worldwide. References: (1) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Professional, Ames, IA, 2006. (2) A. Minuto et al. Phytoparasitica 36:294, 2008. (3) B. A. Summerell et al. Plant Dis. 87:117, 2003. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.


Sample preincubation strategy for sensitive and quantitative detection of clenbuterol in swine urine using a fluorescent microsphere-based immunochromatographic assay.
We describe an ultrasensitive and quantitative immunochromatographic assay to determine the amount of clenbuterol (CLB) in swine urine. In this study, fluorescein isothiocyanate polystyrene fluorescent microspheres were used as probes. A sample preincubation strategy was introduced to this immunochromatographic assay. Results showed that the strategy evidently improved the sensitivity and accuracy of lateral flow assay. The method was completed in 20 min, and a half-maximal inhibitory concentration of 0.13 μg liter(-1) was obtained. The limit of detection of the proposed method to determine CLB in swine urine was 0.01 μg liter(-1), which was lower than the limit of detection of immunochromatographic assays without preincubation. Intra- and interday recoveries of spiked swine urine ranged from 85.0 to 107.5%. The relative standard deviation values of the preincubated test strip ranged from 2.7 to 12.5%. Analysis of the CLB in swine urine samples showed that the result obtained from the lateral flow assay is consistent with that obtained from a commercial enzyme-linked immunosorbent assay kit. Our results suggest that the developed fluorescent microsphere-based immunochromatographic assay may be useful as a rapid screening method to detect CLB quantitatively.


Long-term genetic monitoring reveals contrasting changes in the genetic composition of newly established populations of the intertidal snail Bembicium vittatum.
Newly established populations are susceptible to founder events that reduce genetic variation. This may be counterbalanced by gene flow after populations become established or founders coming from genetically different populations. However, initial gains in genetic diversity may be short-lived if there is limited mixing between lineages and subsequent inbreeding, or if one lineage sweeps to fixation through selection or genetic drift. Here, we report on the genetic changes taking place within two newly established populations of intertidal snail over a 15-year period (∼ 10 generations). Each translocation was set up using multiple, genetically distinct source populations. Our data show that higher levels of variation in the translocated populations compared to the source populations were maintained over time for both nuclear (microsatellite) and mitochondrial genes. Small changes in allele and haplotype frequencies were observed in the source populations and in one of the translocated populations, but marked changes were evident in the other, where there was a dramatic shift towards the genetic make-up of one of the source populations. These genetic changes occurred despite relatively large numbers of founders (200-374 adults) and no evidence of the population experiencing a severe reduction in effective population size. Our study shows that the genetic composition of newly established populations can vary greatly over time and that genetic outcomes can be highly variable, and significantly different from initial expectations, even when they are established using high numbers of individuals and involve source populations from the same geographic regions.


Effect of sodium zirconium cyclosilicate on potassium lowering for 28 days among outpatients with hyperkalemia: the HARMONIZE randomized clinical trial.
Hyperkalemia is a common electrolyte abnormality that may be difficult to manage because of a lack of effective therapies. Sodium zirconium cyclosilicate is a nonabsorbed cation exchanger that selectively binds potassium in the intestine. To evaluate the efficacy and safety of zirconium cyclosilicate for 28 days in patients with hyperkalemia. HARMONIZE was a phase 3, multicenter, randomized, double-blind, placebo-controlled trial evaluating zirconium cyclosilicate in outpatients with hyperkalemia (serum potassium ≥5.1 mEq/L) recruited from 44 sites in the United States, Australia, and South Africa (March-August 2014). Patients (n = 258) received 10 g of zirconium cyclosilicate 3 times daily in the initial 48-hour open-label phase. Patients (n = 237) achieving normokalemia (3.5-5.0 mEq/L) were then randomized to receive zirconium cyclosilicate, 5 g (n = 45 patients), 10 g (n = 51), or 15 g (n = 56), or placebo (n = 85) daily for 28 days. The primary end point was mean serum potassium level in each zirconium cyclosilicate group vs placebo during days 8-29 of the randomized phase. In the open-label phase, serum potassium levels declined from 5.6 mEq/L at baseline to 4.5 mEq/L at 48 hours. Median time to normalization was 2.2 hours, with 84% of patients (95% CI, 79%-88%) achieving normokalemia by 24 hours and 98% (95% CI, 96%-99%) by 48 hours. In the randomized phase, serum potassium was significantly lower during days 8-29 with all 3 zirconium cyclosilicate doses vs placebo (4.8 mEq/L [95% CI, 4.6-4.9], 4.5 mEq/L [95% CI, 4.4-4.6], and 4.4 mEq/L [95% CI, 4.3-4.5] for 5 g, 10 g, and 15 g; 5.1 mEq/L [95% CI, 5.0-5.2] for placebo; P < .001 for all comparisons). The proportion of patients with mean potassium <5.1 mEq/L during days 8-29 was significantly higher in all zirconium cyclosilicate groups vs placebo (36/45 [80%], 45/50 [90%], and 51/54 [94%] for the 5-g, 10-g, and 15-g groups, vs 38/82 [46%] with placebo; P < .001 for each dose vs placebo). Adverse events were comparable between zirconium cyclosilicate and placebo, although edema was more common in the 15-g group (edema incidence: 2/85 [2%], 1/45 [2%], 3/51 [6%], and 8/56 [14%] patients in the placebo, 5-g, 10-g, and 15-g groups). Hypokalemia developed in 5/51 (10%) and 6/56 patients (11%) in the 10-g and 15-g zirconium cyclosilicate groups, vs none in the 5-g or placebo groups. Among outpatients with hyperkalemia, open-label sodium zirconium cyclosilicate reduced serum potassium to normal levels within 48 hours; compared with placebo, all 3 doses of zirconium cyclosilicate resulted in lower potassium levels and a higher proportion of patients with normal potassium levels for up to 28 days. Further studies are needed to evaluate the efficacy and safety of zirconium cyclosilicate beyond 4 weeks and to assess long-term clinical outcomes. clinicaltrials.gov Identifier: NCT02088073.


Cardiac-specific overexpression of mouse cardiac calsequestrin is associated with depressed cardiovascular function and hypertrophy in transgenic mice.
Calsequestrin is a high capacity Ca2+-binding protein in the sarcoplasmic reticulum (SR) lumen. To elucidate the functional role of calsequestrin in vivo, transgenic mice were generated that overexpressed mouse cardiac calsequestrin in the heart. Overexpression (20-fold) of calsequestrin was associated with cardiac hypertrophy and induction of a fetal gene expression program. Isolated transgenic cardiomyocytes exhibited diminished shortening fraction (46%), shortening rate (60%), and relengthening rate (60%). The Ca2+ transient amplitude was also depressed (45%), although the SR Ca2+ storage capacity was augmented, as suggested by caffeine application studies. These alterations were associated with a decrease in L-type Ca2+ current density and prolongation of this channel's inactivation kinetics without changes in Na+-Ca2+ exchanger current density. Furthermore, there were increases in protein levels of SR Ca2+-ATPase, phospholamban, and calreticulin and decreases in FKBP12, without alterations in ryanodine receptor, junctin, and triadin levels in transgenic hearts. Left ventricular function analysis in Langendorff perfused hearts and closed-chest anesthetized mice also indicated depressed rates of contraction and relaxation of transgenic hearts. These findings suggest that calsequestrin overexpression is associated with increases in SR Ca2+ capacity, but decreases in Ca2+-induced SR Ca2+ release, leading to depressed contractility in the mammalian heart.


Evaluating the association between keratoconus and the corneal thickness genes in an independent Australian population.
A recent genome-wide association study (GWAS) identified six loci associated with central corneal thickness that also conferred associated risk of keratoconus (KC). We aimed to assess whether genetic associations existed for these loci with KC or corneal curvature in an independent cohort of European ancestry. In total, 157 patients with KC were recruited from public and private clinics in Melbourne, Australia, and 673 individuals without KC were identified through the Genes in Myopia study from Australia. The following six single-nucleotide polymorphisms (SNPs) that showed a statistically significant association with KC in a recent GWAS study were selected for genotyping in our cohort: rs4894535 (FNDC3B), rs1324183 (MPDZ-NF1B), rs1536482 (RXRA-COL5A1), rs7044529 (COL5A), rs2721051 (FOXO1), and rs9938149 (BANP-ZNF469). The SNPs were assessed for their association with KC or corneal curvature using logistic or linear regression methods, with age and sex included as covariates. Bonferroni corrections were applied to account for multiple testing. Genotyping data were available for five of the SNPs. Statistically significant associations with KC were found for the SNPs rs1324183 (P = 0.001; odds ratio [OR], 1.68) and rs9938149 (P = 0.010; OR, 1.47). Meta-analysis of previous studies yielded genome-wide significant evidence of an association for rs1324183, firmly establishing it as a KC risk variant. None of the SNPs were significantly associated with corneal curvature. The SNPs rs1324183 in the MPDZ-NF1B gene and rs9938149 (between BANP and ZNF4659) were associated with KC in this independent cohort, but their association was via a non-corneal curvature route.


Protein coingestion with alcohol following strenuous exercise attenuates alcohol-induced intramyocellular apoptosis and inhibition of autophagy.
Alcohol ingestion decreases postexercise rates of muscle protein synthesis, but the mechanism(s) (e.g., increased protein breakdown) underlying this observation is unknown. Autophagy is an intracellular "recycling" system required for homeostatic substrate and organelle turnover; its dysregulation may provoke apoptosis and lead to muscle atrophy. We investigated the acute effects of alcohol ingestion on autophagic cell signaling responses to a bout of concurrent (combined resistance- and endurance-based) exercise. In a randomized crossover design, eight physically active males completed three experimental trials of concurrent exercise with either postexercise ingestion of alcohol and carbohydrate (12 ± 2 standard drinks; ALC-CHO), energy-matched alcohol and protein (ALC-PRO), or protein (PRO) only. Muscle biopsies were taken at rest and 2 and 8 h postexercise. Select autophagy-related gene (Atg) proteins decreased compared with rest with ALC-CHO (P < 0.05) but not ALC-PRO. There were parallel increases (P < 0.05) in p62 and PINK1 commensurate with a reduction in BNIP3 content, indicating a diminished capacity for mitochondria-specific autophagy (mitophagy) when alcohol and carbohydrate were coingested. DNA fragmentation increased in both alcohol conditions (P < 0.05); however, nuclear AIF accumulation preceded this apoptotic response with ALC-CHO only (P < 0.05). In contrast, increases in the nuclear content of p53, TFEB, and PGC-1α in ALC-PRO were accompanied by markers of mitochondrial biogenesis at the transcriptional (Tfam, SCO2, and NRF-1) and translational (COX-IV, ATPAF1, and VDAC1) level (P < 0.05). We conclude that alcohol ingestion following exercise triggers apoptosis, whereas the anabolic properties of protein coingestion may stimulate mitochondrial biogenesis to protect cellular homeostasis.


Do open-access dermatology articles have higher citation counts than those with subscription-based access?
Open-access (OA) publishing is increasingly prevalent in dermatology, and many journals now offer hybrid options, including conventional (subscription-based access [SA]) publishing or OA (with an author publishing charge) in a subscription journal. OA publishing has been noted in many disciplines, but this has been rarely studied in dermatology. Using the Clarivate Journal Citation Report, we compiled a list of English-language dermatology hybrid OA journals containing more than 5% OA articles. We sampled any OA review or original research article in 4 issues from 2018 to 2019 and matched an equal number of SA articles. Citation count, citation count excluding self-citations and view counts found using Scopus and Altmetrics score were recorded for each article. Statistical analyses were performed using logistic and negative binomial models using R software. Twenty-seven hybrid dermatology journals were found, and 538 articles were sampled (269 OA, 269 SA). For both original research and review articles, OA articles had significantly higher mean citation counts (mean 13.2, standard deviation [SD] 17.0) compared to SA articles (mean 7.9, SD 8.8) (odds ratio [OR] 1.04; 95% CI 1.02-1.05; P < .001) including when adjusted for time from publication. Original research OA articles had significantly higher citation counts than original research SA articles (excluding self-citations; OR, 1.03; 95% CI, 1.01-1.05; P = .003), and review articles also had OA citation advantage than review SA articles (OR, 1.06; 95% CI, 1.02-1.11; P = .008). There was, however, no significant difference in citation counts between review articles and original research articles (OR, 1.00; 95% CI, 0.19-5.31; P = 1.000). There was no significant difference seen in view counts (OA: mean±SD 17.7±10.8; SA: mean±SD 17.1±12.4) and Altmetric score (OA: mean±SD 13.2±47.8; SA: mean±SD 6.3±25.0) between OA and SA articles. Potential confounders included the fact that more OA articles were published in Europe than in Asia, and pharmaceutical-funded articles were more likely to be published OA. We noted a higher citation count for OA articles than SA articles in dermatology hybrid journals. However, dermatology researchers should take into account confounding factors when deciding whether to increase the impact of their work by selecting OA over SA publishing.


Biodegradable nanoparticles for direct or two-step tumor immunotargeting.
In this study, selective cancer cell targeting of biodegradable poly(lactic acid) (PLA) nanoparticles (NPs) has been investigated in vitro. SKOV-3 (HER2 positive) ovarian cancer and Daudi (CD20 positive) lymphoma cell targeting was mediated by anti-HER2 (trastuzumab, Herceptin) and anti-CD20 (rituximab, Mabthera) monoclonal antibodies (mAbs), respectively. The mAb against nonexpressed antigen serving on each cell as isotype matched irrelevant control. Two different targeting approaches have been studied, a direct method using antibody-labeled NPs (mAb-NPs) and a pretargeting method using the avidin-biotin technology. For the direct protocol, fluorescent PLA-NPs were prepared including 10% 1-pyrenebutanol (PB)-labeled PLA in the NP-preparation (PB-NP). Thiol groups were covalently bound to the PB-NP, and the resulting thiolated PB-NP were coupled with the two mAbs using a bifunctional cross-linker. The effective targeting of cells by mAb-PB-NP was shown by flow cytometry analysis. Clearly anti-HER2-PB-NP specifically bound to the SKOV-3 cells and not to the Daudi cells, while anti-CD20-PB-NPs bound to Daudi cells but not to SKOV-3 cells. Specific mAb-PB-NP binding to tumor cells produced a mean 10-fold or higher signal increase compared to irrelevant IgG-PB-NPs. For the pretargeting protocol, plain PLA-NPs were also thiolated and NeutrAvidin-Rhodamine Red-X (NAR) coupled to the functionalized PLA-NPs with sulfo-MBS. The two-step method was evaluated in vitro by incubating SKOV-3 cells first with biotinylated mAbs followed by NAR-NPs. The relative fluorescence associated to the specific binding of NPs produced a 6-fold increase in flow cytometry signal compared to nonspecific binding. In conclusion, these experiments have shown that NPs covalently coupled with antibodies or NAR can specifically and efficiently bind to cancer cells in both a pretargeting and a direct approach, suggesting that functionalized NPs may be a useful drug carrier for tumor targeting.


Photochemical strategies for the green synthesis of ultrathin Au nanosheets using photoinduced free radical generation and their catalytic properties.
Two-dimensional gold nanosheets represent a class of materials with excellent chemical and structural properties, which are often prepared using a template or toxic CO in organic solvents. Here, we report methylene blue (MB) radicals as a reducing agent to grow freestanding hexagonal ultrathin Au nanosheets with well-tuned thicknesses in water. This is the first time that carbon organic radicals have been used as a reducing agent in metal nanosheet synthesis. Notably, no template is used throughout the synthesis process, and the yield of Au nanosheets is very high. It is found that MB is decisive in the growth of Au nanosheets because no Au nanosheets are obtained in the absence of MB with the same reaction parameters. The resulting nanosheets exhibit excellent catalytic activity during H2O2 decomposition to generate nontoxic O2. Thus, folic acid-conjugated oxygen generating nanosheets could detect cancer cells in serum samples with high sensitivity through pressure signals. Furthermore, the nanosheets exhibit highly efficient activity and selectivity toward the hydrogenation of α,β-unsaturated aldehydes. We anticipate that using MB radicals for the high-yield synthesis of 2D materials in this unique system has demonstrated their effectiveness and provides a green alternative route for producing other 2D nanomaterials.


Relative hopane content confirming the mineral origin of hydrocarbons contaminating foods and human milk.
Hopanes, triterpenoid hydrocarbons formed under geological conditions, were analysed to confirm the mineral origin of the unresolved complex mixtures of hydrocarbons observed in the gas chromatography with flame ionization detection chromatograms of human milk and certain foodstuffs. The 'relative hopane content' (RHC) is introduced, i.e. it is the area ratio of the sum of the hopanes and the paraffins in the same segment of the chromatogram. The RHC in various mineral oil products (motor oils, hydraulic oils, lubricating oils, Vaseline) was 3.4%, with a relative standard deviation of 19%. The RHC determined in samples of vegetable oils, mussels and clams as well as of human milk containing an unresolved complex mixture of hydrocarbons was in the same range, confirming that these samples were contaminated by mineral oil material.


Total Number of Sets as a Training Volume Quantification Method for Muscle Hypertrophy: A Systematic Review.
Baz-Valle, E, Fontes-Villalba, M, and Santos-Concejero, J. Total number of sets as a training volume quantification method for muscle hypertrophy: A systematic review. J Strength Cond Res 35(3): 870-878, 2021-This review aimed to determine whether assessing the total number of sets is a valid method to quantify training volume in the context of hypertrophy training. A literature search on 2 databases (PubMed and Scopus) was conducted on May 18, 2018. After analyzing 2,585 resultant articles, studies were included if they met the following criteria: (a) studies were randomized controlled trials, (b) studies compared the total number of sets, repetition range, or training frequency, (c) interventions lasted at least 6 weeks, (d) subjects had a minimum of 1 year of resistance training experience, (e) subjects' age ranged from 18 to 35 years, (f) studies reported morphologic changes through direct or indirect assessment methods, (g) studies involved subjects with no known medical conditions, and (h) studies were published in peer-reviewed journals. Fourteen studies met the inclusion criteria. According to the results of this review, the total number of sets to failure, or near to, seems to be an adequate method to quantify training volume when the repetition range lies between 6 and 20+ if all the other variables are kept constant. This approach requires further development to assess whether specific numbers of sets are key to inducing optimal muscle gains.


PAQR3 modulates cholesterol homeostasis by anchoring Scap/SREBP complex to the Golgi apparatus.
Cholesterol biosynthesis is regulated by transcription factors SREBPs and their escort protein Scap. On sterol depletion, Scap/SREBP complex is transported from endoplasmic reticulum (ER) to the Golgi apparatus where SREBP is activated. Under cholesterol sufficient condition, Insigs act as anchor proteins to retain Scap/SREBP in the ER. However, the anchor protein of Scap/SREBP in the Golgi is unknown. Here we report that a Golgi-localized membrane protein progestin and adipoQ receptors 3 (PAQR3) interacts with Scap and SREBP and tethers them to the Golgi. PAQR3 promotes Scap/SREBP complex formation, potentiates SREBP processing and enhances lipid synthesis. The mutually exclusive interaction between Scap and PAQR3 or Insig-1 is regulated by cholesterol level. PAQR3 knockdown in liver blunts SREBP pathway and decreases hepatic cholesterol content. Disrupting the interaction of PAQR3 with Scap/SREBP by a synthetic peptide inhibits SREBP processing and activation. Thus, PAQR3 regulates cholesterol homeostasis by anchoring Scap/SREBP to the Golgi and disruption of such function reduces cholesterol biosynthesis.


Postoperative duplex scan surveillance of axillofemoral bypass grafts.
Duplex scan surveillance (DS) for axillofemoral bypass grafts (AxFBGs) has not been extensively studied. The intent of this study was twofold: 1, to characterize the flow velocities within AxFBGs; and 2, to determine whether postoperative DS is useful in assessment of future patency of AxFBGs. We identified all patients who underwent AxFBG procedures between January 1996 and January 2001 at our combined university and Veterans Affairs hospital vascular surgical service. All grafts were performed with ringed 8-mm polytetrafluoroethylene with the distal limb of the axillofemoral component anastomosed to the hood of the femoral-femoral graft. DS was every 3 months for 1 year and every 6 months thereafter. Duplex scan results were compared in primarily patent grafts with grafts that thrombosed. Graft failures from infection were excluded. Influences of ankle-brachial index, blood pressure, outflow patency, operative indication, and comorbidities on graft patency were analyzed. One hundred twenty patients underwent AxFBG procedures. Twenty-eight were excluded because of infection or death before surveillance examination. Fourteen were lost to follow-up, 23 had failed grafts from occlusion, and 55 had grafts that remained patent. In the 78 patients evaluated during long-term follow-up period, the mean peak systolic velocities (PSVs) at the proximal (axillary) anastomosis during the first postoperative year ranged from 153 to 194 cm/s. Mean PSVs at the mid portion of the axillofemoral graft during the first postoperative year ranged from 100 to 125 cm/s, whereas those for the distal axillofemoral anastomosis ranged from 93 to 129 cm/s. Mean midgraft and distal anastomotic velocities obtained before thrombosis were significantly lower in the thrombosed grafts compared with the last recorded velocities at the same sites in the patent grafts (mean PSV, 84 versus 112 cm/s; P =.015; mean PSV, 89 versus 127 cm/s; P =.024, respectively). Forty-eight percent of occluded grafts had a mean midgraft PSV at last observation of less than 80 cm/s. Blood pressure correlated with midgraft velocity (r = 0.415; P <.05). With multivariate logistic regression analysis, a mean midgraft velocity less than 80 cm/s was the sole independent factor associated with graft failure (P <.01). No patients with midgraft velocities greater than 155 cm/s had occlusion. Flow velocity varies widely within and among AxFBGs. Patency of AxFBGs is associated with higher midgraft PSV, and thrombosis with midgraft velocities less than 80 cm/s.