Cryopreservation of Plasmodium Sporozoites.
Malaria is a deadly disease caused by the parasite, Plasmodium, and impacts the lives of millions of people around the world. Following inoculation into mammalian hosts by infected mosquitoes, the sporozoite stage of Plasmodium undergoes obligate development in the liver before infecting erythrocytes and causing clinical malaria. The most promising vaccine candidates for malaria rely on the use of attenuated live sporozoites to induce protective immune responses. The scope of widespread testing or clinical use of such vaccines is limited by the absence of efficient, reliable, or transparent strategies for the long-term preservation of live sporozoites. Here we outline a method to cryopreserve the sporozoites of various human and murine Plasmodium species. We found that the structural integrity, viability, and in vivo or in vitro infectiousness were conserved in the recovered cryopreserved sporozoites. Cryopreservation using our approach also retained the transgenic properties of sporozoites and immunization with cryopreserved radiation attenuated sporozoites (RAS) elicited strong immune responses. Our work offers a reliable protocol for the long-term storage and recovery of human and murine Plasmodium sporozoites and lays the groundwork for the widespread use of live sporozoites for research and clinical applications.


Measuring Impatience in Intertemporal Choice.
In general terms, decreasing impatience means decreasing discount rates. This property has been usually referred to as hyperbolic discounting, although there are other discount functions which also exhibit decreasing discount rates. This paper focuses on the measurement of the impatience associated with a discount function with the aim of establishing a methodology to compare this characteristic for two different discount functions. In this way, first we define the patience associated with a discount function in an interval as its corresponding discount factor and consequently we deduce that the impatience at a given moment is the corresponding instantaneous discount rate. Second we compare the degree of impatience of discount functions belonging to the same or different families, by considering the cases in which the functions do or do not intersect.


CDKL5 mutations as a cause of severe epilepsy in infancy: clinical and electroencephalographic long-term course in 4 patients.
CDKL5 mutations cause severe epilepsy in infancy with subsequent epileptic encephalopathy. As yet, few studies report on long-term observations in patients with CDKL5-related epileptic encephalopathy. In this study, we describe the evolution of the epilepsy phenotype and the electroencephalographic (EEG) features in 4 patients during a maximum observation period of 22 years. All 4 patients had epilepsy starting with focal seizures in the first 3 months of life, evolving to epileptic spasms between the ages of 2 and 6 years and later on to tonic seizures. In 3 patients, epilepsy was resistant to antiepileptic therapy. Although there was no common EEG pattern in all patients, late hypsarrhythmia until the age of 9 years was observed in 2 patients. CDKL5-related epileptic encephalopathies are a group of refractory seizure disorders starting in early infancy. The phenomenon of late hypsarrhythmia may help define a subgroup of patients with severe and adverse outcomes.


Big Is Beautiful: Enhanced saRNA Delivery and Immunogenicity by a Higher Molecular Weight, Bioreducible, Cationic Polymer.
Self-amplifying RNA (saRNA) vaccines are highly advantageous, as they result in enhanced protein expression compared to mRNA (mRNA), thus minimizing the required dose. However, previous delivery strategies were optimized for siRNA or mRNA and do not necessarily deliver saRNA efficiently due to structural differences of these RNAs, thus motivating the development of saRNA delivery platforms. Here, we engineer a bioreducible, linear, cationic polymer called "pABOL" for saRNA delivery and show that increasing its molecular weight enhances delivery both in vitro and in vivo. We demonstrate that pABOL enhances protein expression and cellular uptake via both intramuscular and intradermal injection compared to commercially available polymers in vivo and that intramuscular injection confers complete protection against influenza challenge. Due to the scalability of polymer synthesis and ease of formulation preparation, we anticipate that this polymer is highly clinically translatable as a delivery vehicle for saRNA for both vaccines and therapeutics.


Marketing of breast-milk substitutes in Zambia: evaluation of compliance to the international regulatory code.
We sought to assess the level of non-compliance with the International Code of Marketing breast-milk substitutes (BMS) and/or Statutory Instrument (SI) Number 48 of 2006 of the Laws of Zambia in two suburbs, Kalingalinga and Chelstone, in Zambia. This was a cross sectional survey. Shop owners (80), health workers (8) and mothers (214) were interviewed. BMS labels and advertisements (62) were observed. The primary outcome was mean non-compliance defined as the number of article violations divided by the total 'obtainable' violations. The score ranges from 0 to 1 with 0 representing no violations in all the articles and one representing violations in all the articles. A total of 62 BMS were assessed. The mean non-compliance score by manufacturers in terms of violations in labelling of BMS was 0.33 (SD = 0.28; 95% CI: 0.26, 0.40). These violations were mainly due to labels containing pictures or graphics representing an infant. 80 shops were also assessed with mean non-compliance score in respect of violations in tie-in-sales, special display, and contact with mothers at the shop estimated as 0.14 (SD = 0.14; 95% CI: 0.11, 0.18). Non-compliance with the Code and/or the local SI is high after 10 years of domesticating the Code.


Cooking Methods for a Soft Diet Using Chicken Based on Food Texture Analysis.
Undernutrition caused by difficulties in masticating is of growing concern among the elderly. Soft diets are often served at nursing homes; however, the styles differ with nursing homes. Improperly modified food texture and consistency may lead to further loss of nutritive value. Therefore, we developed a method to produce a soft diet using chicken. The texture-modified chicken was prepared by boiling a mixture of minced chicken and additive foodstuff that softened the meat. The best food additive was determined through testing cooking process, size after modification and texture. The optimum proportions of each component in the mixture were determined measuring food texture using a creep meter. Teriyaki chicken was cooked using the texture-modified chicken, and provided to a nursing home. The amount of food intake by elderly residents was subsequently surveyed. This study involved 22 residents (1 man and 21 women; mean age 91.4±5.3 y). Consequently, yakifu, which was made from wheat gluten, was the most suitable additive foodstuff. The hardness of the texture-modified chicken, with proportions of minced chicken, yakifu, and water being 50%, 10%, and 40% respectively, was under 40,000 N/m2. The intake amount of the texture-modified chicken of subjects whose intake amount of conventional chicken using chicken thigh was not 100% was significantly higher. These findings suggest that properly modified food textures could contribute to improve the quality of meals by preventing undernutrition among the elderly with mastication difficulties.


Evolution of Validated Biomarkers and Intraoperative Parameters in the Development of Postoperative ARDS.
Patients who develop ARDS from medical or traumatic causes typically present after the inciting event has already occurred. Postoperative ARDS is unique in that the inciting insult potentially responsible for ARDS is known ahead of time, which provides an opportunity to study the early pathophysiology of ARDS. The objective of this study was to better understand the early pathophysiology of postoperative ARDS through a temporal analysis of key biomarkers of interest. We performed a case-control study of adults undergoing elective thoracic, aortic vascular, or cardiac surgery, which placed them at increased risk of developing postoperative ARDS. Biomarkers were measured at baseline, 2 h, and 6 h after the key intraoperative event believed to be responsible for ARDS. Of the 467 subjects enrolled, 26 developed ARDS and were matched to non-ARDS controls 1:2 based on age, sex, surgical procedure, and surgical lung injury prediction score. Patients with ARDS were more likely to have lower preoperative albumin (P = .029), longer surgery (P = .007), larger amounts of intraoperative fluid (P = .036), and higher intraoperative peak inspiratory pressures (P = .006). Baseline plasminogen activator inhibitor-1 levels were higher in the ARDS group (P = .03). Changes in postoperative biomarker levels from baseline were greater in the ARDS group for interleukin-8 (baseline to 6 h, P = .02) and surfactant protein-D (baseline to 2 h, P = .009). Our study supported the hypothesis that dysregulated coagulation, inflammation, and epithelial injury are pathophysiologic features of early postoperative ARDS. Interleukin-8, plasminogen activator-1, and surfactant protein-D may help predict development of postoperative ARDS.


Trail Making Test A improves performance characteristics of the International HIV Dementia Scale to identify symptomatic HAND.
Although HIV-associated dementia (HAD) occurs in less than 5 % of individuals with access to combination antiretroviral therapy, rates of milder forms of HIV-associated neurocognitive disorder (HAND) are much higher. We sought to define an optimal cut point for the International HIV Dementia Scale (IHDS) in Thailand for the identification of symptomatic HAND, defined as both HAD and mild neurocognitive disorder. We then sought to determine if adding a simple test from a larger neuropsychological battery could improve the performance characteristics for identifying symptomatic HAND. In this study, subjects comprising 75 seropositive adults in Bangkok, Thailand, completed neuropsychological tests and underwent a full neurological assessment. HAND diagnoses were determined by consensus conference using the 2007 Frascati criteria, blinded to the IHDS results. The optimal IHDS cut point was determined by receiver operating characteristic analysis with cross-validation. Individual neuropsychological tests were then evaluated and combined with the IHDS to test performance characteristics. The IHDS was poor at detecting symptomatic HAND at the optimized cut point of ≤ 10 (sensitivity, 53.3 %; specificity, 89.8 %). Trail Making Test A was most effective in improving performance characteristics when combined with the IHDS, with net sensitivity of 86 % and specificity of 79 %. In this setting, the IHDS performed poorly in identifying symptomatic HAND, but was substantially improved by the addition of Trail Making Test A, which typically requires less than 2 min to complete. This combination should be validated in a larger setting since it may address the critical need for HAND screening instruments in international settings.


Regeneration of rete ridges in Lanyu pig (Sus scrofa): Insights for human skin wound healing.
Rete ridges are important to the mechanical function of skin in animals with minimal hair, including humans. As mice do not exhibit rete ridges, the need for a quality animal model is pertinent. Here, we develop a Lanyu pig (Sus scrofa) full-thickness wound model to explore tissue regeneration because the architecture and function are similar to humans and inbred genetic variants are available. Full- and partial-thickness wounds were generated on the dorsum. Full-thickness wounds at post-wound day 57 exhibit severe scar with no signs of wound-induced hair follicle neogenesis. Wound contraction is greater in the anterior/posterior relative to the medial/lateral axis. In wound beds, K14+ cells increased while K10+ , p63+ and PCNA+ cells decreased compared to unwounded tissue. Epithelial β-catenin is unchanged. The wound bed expresses more ColI, less ColIII and no elastin. Rete ridges do not form after full-thickness wounding, but incompletely regenerate after partial-thickness wounding. An alkaline phosphatase (ALP)+ cell population, not associated with hair follicles, is present at the bottom of the rete ridge basal layer in pig and human unwounded skin. These K5+ /K10- /PCNA- /ALP+ epithelial cells are absent after full-thickness wounding but reappear after partial-thickness wounding, before invagination of new rete ridges. In summary, full-thickness wounding on the dorsum of Lanyu pigs results in scar formation and perturbed molecular expression while partial-thickness wounding permits limited rete ridge and papillary dermis regeneration. Future functional studies and further characterization will help contribute knowledge for the regenerative medicine field.


It's not easy being green: the viridans group streptococci, with a focus on pediatric clinical manifestations.
The viridans group streptococci (VGS) are a heterogeneous group of organisms that can be human commensals, colonizing the gastrointestinal and genitourinary tracts in addition to the oral mucosa. VGS are generally considered to be of low pathogenic potential in immunocompetent individuals. However, in certain patient populations, VGS can cause invasive disease, such as endocarditis, intra-abdominal infection, and shock. Within the VGS, the rates and patterns of antimicrobial resistance vary greatly depending upon the species identification and the patient population. In general, Streptococcus mitis group organisms are resistant to more antimicrobial agents than the other VGS species. This review addresses current VGS taxonomy, in addition to the current methodologies being used in clinical microbiology laboratories for identification of VGS. Automated systems struggle overall with species level identification and susceptibility testing for VGS. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) identification is emerging as a potential alternative for organism identification. A review of recent pediatric-specific data regarding the clinical manifestations of VGS revealed that the Streptococcus anginosus group (SAG) organisms may be important pathogens in pediatric patients and that the VGS may contribute to disease in patients with cystic fibrosis. It also appears that rates of antimicrobial resistance in VGS in pediatric patients are surpassing those of the adult population.


Hereditary pituitary dwarfism in mice affects skeletal and cardiac myosin isozyme transitions differently.
The dwarf mutation in mice interferes with the development of those anterior pituitary cells responsible for production of thyroid stimulating hormone, growth hormone, and prolactin. Myosin isozyme transitions in both cardiac and skeletal muscle were also found to be affected in this mutant. Electrophoresis of native myosins demonstrated that the fetal (V3) to adult (V1) ventricular cardiac isozyme transition was completely blocked in dwarf mice; in contrast, the neonatal to adult fast myosin transition in hind limb skeletal muscle was slowed but not totally inhibited. The persistence of neonatal myosin heavy chain for up to 55-75 d after birth in dwarf mice, as compared with 16 d in normal mice, was directly demonstrated by polypeptide and immunopolypeptide mapping. Morphological examination of 18-36-d-old dwarf skeletal muscles by optical and electron microscopy revealed a relative immaturity, but no signs of gross pathology were evident. Immunocytochemical analysis showed that the abnormal persistence of neonatal myosin occurs in most of the fibers. Multiple injections of thyroxine restored a normal isozyme complement to both cardiac and skeletal muscles within 11-15 d. Therefore, the effects of the dwarf mutation on myosin isozymes can be explained by the lack of thyroid hormone in these animals. Because the synthesis of growth hormone is not stimulated by thyroid hormone in dwarf mice as it would be in normal animals, these results demonstrate that thyroid hormone promotes myosin isozyme transitions independent of growth hormone production.


Study on the mechanism of interference between Friend leukemia and Sindbis viruses in tissue culture.
Some mechanisms of interference developing in BALB/c mouse embryo fibroblast culture (MEC) infected with Friend leukemia virus (FLV) and 48 hours later superinfected with Sindbis virus (SV) was studied. In FLV-infected cells the amount of SV antigen formed was 2-3 times lower than in SV monoinfection, as indicated by immunofluorescence and cytofluorimetry. Electron microscopic examination showed that in mixed infection the number of newly formed SV particles decreased markedly (by 90%) despite the presence of compact aggregates of viral nucleocapsids in the cytoplasm. When the cells were initially infected with arbovirus and then superinfected with FLV, formation of virus antigen and virions of both viruses was not disturbed. Pre-treatment of cell monolayer with dactinomycin (0.2 mug/ml) blocked interferon production in MEC culture and inhibited interference between FLV and SV. It is assumed that interference between FLV and SV is associated with known mechanisms of interferon action as well as with disturbance of the stage of SV particles assembly and their release from the cell. Due to incomplete cycle of SV reproduction interrupted at the stage of ribonucleoprotein formation, productive type of its interaction with MEC cells is disturbed.


Stable Small Composite Microbubbles Decorated with Magnetite Nanoparticles - A Synergistic Effect between Surfactant Molecules and Nanoparticles.
Three approaches to preparing iron oxide nanoparticle-decorated microbubbles (NP-decoMBs) have been investigated. The size and stability characteristics of these microbubbles (MBs) were investigated by optical microscopy, laser light scattering and an acoustical method, and compared with those of non-decorated MBs. First, magnetite nanoparticles (Fe3O4NPs) grafted with dimyristoylphosphatidylcholine (DMPC) were synthesized and used to prepare MBs by brief sonication under an atmosphere of air saturated with perfluorohexane. These MBs had a rather large mean radius (r ~ 12 µm), and a moderate volume of encapsulated gas. Remarkably, a second approach that consisted of dispersing unbound DMPC molecules in the aqueous phase along with DMPC-grafted Fe3O4NPs prior to sonication was found to drastically change the situation, allowing the obtaining of monomodal populations of much smaller (r ~ 0.6 µm) NP-decoMBs. The latter were echogenic and stable for at least 10 days at room temperature, without significant variation of their size characteristics. In a third approach, NP-decoMBs were directly prepared from dispersions of naked Fe3O4NPs in the presence of DMPC. The resulting NP-decoMBs suspensions consisted of broadly distributed bubble populations mostly containing two populations (with r ~ 5 and ~ 15 µm). Control microbubbles made of DMPC only were small (r ~ 1.3 µm), although not as small as those formed from DMPC-grafted Fe3O4NPs in the presence of free DMPC, and were less stable, with a room temperature half-life of only ~1 day. These observations imply that there is a synergy between the Fe3O4NPs and the DMPC molecules in the air/water interfacial film stabilization process.


Hepatorenal syndrome: a critical update.
Acute kidney injury (AKI) is common in cirrhotic patients with ascites. Although not the primary etiology of AKI in cirrhotic patients, hepatorenal syndrome (HRS) is a unique form of AKI that develops only in cirrhotic patients. Intense renal vasoconstriction is the hallmark of HRS. Different mechanisms contribute to renal vasoconstriction, with splanchnic vasodilatation and reduced effective blood volume playing a central role. Diagnostic criteria for HRS have been developed and were recently modified, but diagnosing HRS and differentiating it from other causes of AKI in cirrhotic patients continues to be a difficult task in some patients. Given its overall dismal prognosis, strategies to prevent HRS have been developed and proved to be effective in reducing HRS prevalence among cirrhotic patients. Liver transplantation is the ultimate treatment, but more than one treatment modality can be utilized as a bridge to transplantation. This review provides an update on our current understanding of HRS with emphasis on the underlying pathophysiological mechanisms involved, difficulties in diagnosis, and different treatment modalities.


Growth of Campylobacter pylori in liquid media.
Until recently, broth cultivation techniques for Campylobacter pylori were unavailable. We developed a method to cultivate bacterial cells within 24 h in liquid media. Cultivation in broth depended on the adequate dispersion of appropriate gases. A static broth at 37 degrees C in a GasPak jar (BBL Microbiology Systems, Cockeysville, Md.) with a CampyPak (BBL) envelope did not support growth after 5 days of incubation. A broth placed in a flask on a Gyrotory water bath shaker (150 rpm; New Brunswick Scientific Co., Inc., Edison, N.J.) fitted with a gassing hood connected to a gas mixture of 10% CO2, 5% O2, and 85% N2 supported good growth. An initial inoculum of 10(5), 10(3) to 10(4), or 10(2) CFU/ml resulted in greater than or equal to 10(8) CFU/ml after incubation for 24, 48, or 72 h, respectively. Under these conditions, the bacteria grew as motile, spiral bacilli rather than the oval and coccal bacilli occasionally reported. Several bases supported good growth when supplemented with serum. For the determination of basal growth conditions, brucella broth base was used. Fetal calf serum (1%) provided maximum growth. Vitox was not necessary for growth and did not augment growth. C. pylori grew over a wide optimal pH range of 5.5 to 8.5.


The response of zinc transporter gene expression of selected tissues in a pig model of subclinical zinc deficiency.
This study compared the relative mRNA expression of all mammal zinc (Zn) transporter genes in selected tissues of weaned piglets challenged with short-term subclinical Zn deficiency (SZD). The dietary model involved restrictive feeding (450 g/animal*day-1) of a high-phytate diet (9 g/kg) supplemented with varying amounts of zinc from ZnSO4*7H2O ranging from deficient to sufficient supply levels (total diet Zn: 28.1, 33.6, 38.8, 42.7, 47.5, 58.2, 67.8, 88.0 mg Zn/kg). Total RNA preparations comprised jejunal and colonic mucosa as well as hepatic and nephric tissue. Statistical modelling involved broken-line regression (P≤.05). ZIP10 and ZIP12 mRNAs were not detected in any tissue and ZnT3 mRNA was only identified in the kidney. All other genes were expressed in all tissues but only a few gene expression patterns allowed a significant (P<.0001) fitting of broken-line regression models, indicating homeostatic regulation under the present experimental conditions. Interestingly, these genes could be subcategorized by showing significant turnarounds in their response patterns, either at ~40 or ~60 mg Zn/kg diet (P<.0001). In conclusion, the present study showed clear differences in Zn transporter gene expression in response to SZD compared to the present literature on clinical models. We recognized that certain Zn transporter genes were regulated under the present experimental conditions by two distinct homeostatic networks. For the best of our knowledge, this represents the first comprehensive screening of Zn transporter gene expression in a highly translational model to human physiology.


Personalised Profiling of Innate Immune Memory Induced by Nano-Imaging Particles in Human Monocytes.
Engineered nanoparticles used for medical purposes must meet stringent safety criteria, which include immunosafety, i.e., the inability to activate possibly detrimental immune/inflammatory effects. Even medical nanomaterials devoid of direct immunotoxic or inflammatory effects may have an impact on human health if able to modify innate memory, which is the ability to "prime" future immune responses towards a different, possibly more detrimental reactivity. Although innate memory is usually protective, anomalous innate memory responses may be at the basis of immune pathologies. In this study, we have examined the ability of two nanomaterials commonly used for diagnostic imaging purposes, gold and iron oxide nanoparticles, to induce or modulate innate memory, using an in vitro model based on human primary monocytes. Monocytes were exposed in culture to nanoparticles alone or together with the bacterial agent LPS (priming phase/primary response), then rested for six days (extinction phase), and eventually challenged with LPS (memory/secondary response). The memory response to the LPS challenge was measured as changes in the production of inflammatory (TNFα, IL-6) and anti-inflammatory cytokines (IL-10, IL-1Ra), as compared to unprimed monocytes. The results show that both types of nanoparticles can have an effect in the induction of memory, with changes observed in the cytokine production. By comparing nanomaterials of different shapes (spherical vs. rod-shaped gold particles) and different size (17 vs. 22 nm diameter spherical iron oxide particles), it was evident that innate memory could be differentially induced and modulated depending on size, shape and chemical composition. However, the main finding was that the innate memory effect of the particles was strongly donor-dependent, with monocytes from each donor showing a distinct memory profile upon priming with the same particles, thereby making impossible to draw general conclusions on the particle effects. Thus, in order to predict the effect of imaging nanoparticles on the innate memory of patients, a personalised profiling would be required, able to take in consideration the peculiarities of the individual innate immune reactivity.


Milk and meat fatty acids from sheep fed a plantain-chicory mixture or a grass-based permanent sward.
Plantain and chicory are interesting forage species since they present good nutritional quality and are more resistant to drought than many temperate grasses. The fatty acid (FA) profile in milk and meat is related to a growing concern for the consumption of healthy foods, that is, with a lower content of saturated FA, higher polyunsaturated FA (PUFA) and a favourable n-6 : n-3 FAs ratio. Our objective was to evaluate the FA content in ewe's milk and lamb's meat fed a plantain-chicory mixture (PCH) or a grass-based permanent sward (GBS) dominated by perennial ryegrass. Eighteen Austral ewes in mid-lactation were allocated to PCH and GBS treatments. Milk samples were obtained during September (spring). Thirty weaned lambs were finished on both treatments from November to December (7 weeks), slaughtered and their meat sampled. Fat from milk and meat samples was extracted and stored until analysed by gas chromatography. Milk fat from GBS was higher than from PCH (P < 0.05) in C18:0 (11 385 v. 5874 mg/100 g FA), 9c-18:1 (15 750 v. 8565 mg/100 g FA), 11 t-18:1 (4576 v. 2703 mg/100 g FA) and 9c,11 t-18:2 (1405 v. 921 mg/100 g FA) and lower in 18:2n-6 (827 v. 1529 mg/100 g FA) and 18:3n-3 (943 v. 1318 mg/100 g FA) FA. Total mono-unsaturated FA was higher in GBS than PCH (P < 0.05). Meat fat from PCH swards presented a higher (P < 0.05) content than GBS for 18:2n-6 (46.8 v. 28.2 mg/100 g FA), linolenic (24.6 v. 14.2 mg/100 g FA), polyunsaturated FA (119.7 v. 73.4 mg/100 g FA), n-6 (65.9 v. 40.8 mg/100 g FA) and n-3 (53.8 v. 32.5 mg/100 g FA), respectively. No effect of treatment (P > 0.05) was detected for 9c-18:1 (283.9 v. 205.8 mg/100 g FA), 11 t-18:1 (26.2 v. 19.3 mg/100 g FA) and 9c,11 t-18:2 (10.1 v. 7.6 mg/100 g FA), for PCH and GBS. These results suggest that grazing a PCH mixture results in a higher concentration of PUFA in ewes' milk and in lambs' fat, as compared to a GBS sward.


Host-directed therapies for bacterial and viral infections.
Despite the recent increase in the development of antivirals and antibiotics, antimicrobial resistance and the lack of broad-spectrum virus-targeting drugs are still important issues and additional alternative approaches to treat infectious diseases are urgently needed. Host-directed therapy (HDT) is an emerging approach in the field of anti-infectives. The strategy behind HDT is to interfere with host cell factors that are required by a pathogen for replication or persistence, to enhance protective immune responses against a pathogen, to reduce exacerbated inflammation and to balance immune reactivity at sites of pathology. Although HDTs encompassing interferons are well established for the treatment of chronic viral hepatitis, novel strategies aimed at the functional cure of persistent viral infections and the development of broad-spectrum antivirals against emerging viruses seem to be crucial. In chronic bacterial infections, such as tuberculosis, HDT strategies aim to enhance the antimicrobial activities of phagocytes and to curtail inflammation through interference with soluble factors (such as eicosanoids and cytokines) or cellular factors (such as co-stimulatory molecules). This Review describes current progress in the development of HDTs for viral and bacterial infections, including sepsis, and the challenges in bringing these new approaches to the clinic.


Thiazolidinedione effects on blood pressure in diabetic patients with metabolic syndrome treated with glimepiride.
The aim of our study was to compare the long-term effect of pioglitazone and rosiglitazone on blood pressure control of diabetic patients with metabolic syndrome treated with glimepiride. We evaluated 91 type 2 diabetic patients with metabolic syndrome. All were required to have been diagnosed as diabetic for at least 6 months, and to have failed to achieve glycemic control by dietary changes and the maximum tolerated dose of the oral hypoglycemic agents sulfonylureas or metformin. All patients took a fixed dose of 4 mg/day glimepiride. We administered pioglitazone (15 mg/day) or rosiglitazone (4 mg/day) for 12 months in a randomized, double-blind fashion, and evaluated body mass index (BMI), glycemic control, blood pressure and heart rate (HR) throughout the treatment period. A total of 87 patients completed the study and were randomized to receive double-blind treatment with pioglitazone or rosiglitazone. An increase in BMI was observed after 12 months (p < 0.05) in both groups. After 9 and 12 months, there were significant decreases in glycated hemoglobin (HbA(1c)), mean fasting plasma glucose (FPG), postprandial plasma glucose (PPG), fasting plasma insulin (FPI), and postprandial plasma insulin (PPI) in both treatment groups (p < 0.05 at 9 months and p < 0.01 at 12 months for all parameters). Furthermore, homeostasis model assessment index (HOMA index) improvement was obtained at 9 and 12 months (p < 0.05 and p < 0.01, respectively) in both groups. Significant systolic blood pressure (SBP) and diastolic blood pressure (DBP) improvement (p < 0.05, respectively) was observed in both groups after 12 months. There were no significant changes in transaminases at any point during the study. We can conclude that the association of a thiazolinedione to the glimepiride treatment of type 2 diabetic subjects with metabolic syndrome is associated to a significant improvement in the long-term blood pressure control, related to a reduction in insulin-resistance.


Cancer risk after ABO-incompatible living-donor kidney transplantation.
Recipients of ABO-incompatible (ABOi) living-donor kidney transplants often undergo more intense immunosuppression than their ABO-compatible counterparts. It is unknown if this difference leads to higher cancer risk after transplantation. Single-center studies are too small and lack adequate duration of follow-up to answer this question. We identified 318 ABOi recipients in the Transplant Cancer Match Study, a national linkage between the Scientific Registry of Transplant Recipients and population-based U.S. cancer registries. Seven cancers (non-Hodgkin lymphoma, Merkel cell carcinoma, gastric adenocarcinoma, hepatocellular carcinoma, thyroid cancer, pancreatic cancer, and testicular cancer) were identified among ABOi recipients. We then matched ABOi recipients to ABO-compatible controls by age, gender, race, human leukocyte antigen mismatch, retransplantation, and transplant year. There was no demonstrable association between ABOi and cancer in unadjusted (incidence rate ratio, 0.83; 95% confidence interval, 0.33-1.71; P=0.3) or matched control (incidence rate ratio, 0.99; 95% confidence interval, 0.38-2.23; P=0.5) analyses. To the extent that could be determined in this registry study, current desensitization protocols are not associated with increased risk of cancer after transplantation.


Predictors of death including quality of positive pressure ventilation during newborn resuscitation and the relationship to outcome at seven days in a rural Tanzanian hospital.
Effective positive pressure ventilation (PPV) of non-breathing newborns is crucial in facilitating cardio-respiratory adaptation at birth. Identifying predictors of death in newborns receiving PPV is important in order to facilitate preventative strategies. The objective of this study was to determine the perinatal predictors of death including the quality of PPV administered among admitted newborns. An observational study of admitted newborns who received PPV after birth was conducted. Research assistants observed all deliveries and recorded perinatal events on data collection forms. Measured heart rate (HR) and ventilation parameters were then compared between newborns who died and survivors. Newborns (n = 232) were studied between October 2014 and November 2016. Newborns who died (n = 53) compared to survivors (n = 179) had more fetal heart rate (FHRT) abnormalities (12/53 vs 19/179; p = 0.03); lower initial HR (<100 beats/minute) at start of PPV (44/48 vs 77/139; p<0.001); and a longer time for HR to increase >100 beats/minute from birth (180 vs 149 seconds; p = 0.07). Newborns who died compared to survivors took longer time (14 vs 4 seconds; p = 0.008) and more inflations (7 vs 3; p = 0.006) to achieve an expired volume (Vt) of 6 ml/kg, respectively. Median delivered Vt during the first 60 seconds of PPV was less in newborns who died compared to survivors (5 vs 6 ml/kg; p = 0.12). Newborns who died proceeded to severe encephalopathy (15/31 vs 1/59; p<0.001) compared to survivors. Depressed newborns who proceeded to death compared to survivors, exhibited delayed HR response to PPV which may partly reflect FHRT abnormalities related to interruption of placental blood flow, and/or a timely delay in establishing adequate Vt. Depressed newborns progressed to moderate/severe encephalopathy. Improving FHRT monitoring to identify fetuses at risk for expedited delivery, coupled with optimizing delivery room PPV might decrease mortality in this setting.


Topical Fluoride Applications Related Posts Analysis on Twitter Using Natural Language Processing.
Social media is today a comprehensive source of data that can serve as a guide to professionals in issues related to public health. The purpose of this paper is to investigate the content of topical fluoride-related Twitter posts made over a 3-year period in order to improve our understanding of Twitter users' perceptions and treatment experiences. A continuous cross-sectional sample of Tweets on the subject of 'approaches to the topical fluoride treatment of tooth decay' was collected from the Twitter social networking platform between 1 January 2017 and 1 January 2020 using a software application developed for this research that makes use of the Twitter advanced search API. The words and phrases used for the identification of related Tweets were determined through a screening of the topical fluoride keywords of previous studies, and a search was conducted in the English language. To better arrange the collected Tweets and to make the data more meaningful, firstly one of the natural language process techniques - Tokenization - was applied, after which the Tweets were converted into a set of meaningful words and regular expressions. The Tweets were then compared with each other, word-by-word, with the help of a word-based Levenshtein distance algorithm, after which two experts in the computational social science domain labelled each Tweet. A total of 132,358 Tweeter posts referencing topical fluoride applications were collected, of which 110,847 were eliminated through the use of a word-based Levenshtein distance algorithm, and the remaining corpus of 21,511 posts was analysed and evaluated for specific content. Within the garnered data, 48.5% (n = 10,428) of the Tweeter posts concerned topical fluoride treatments, and 7% (n = 1,507) reported experiences with topical fluoride treatment. Negative Tweeter posts about topical fluoride treatment (5,679, 26.4%) vastly outnumbered those that were positive (3,897, 18.1%). The current study achieved its main objectives of analysing topical fluoride application-related posts made on social media. From the garnered Twitter data, it can be understood that Twitter users regularly share their concerns and negative sentiments about the side effects of topical fluoride treatments on the platform. Future explorations of social media may aid public health and dental professionals in the development of strategies to educate the public and to raise awareness of the importance of topical fluoride applications.


PROVENGE (Sipuleucel-T) in prostate cancer: the first FDA-approved therapeutic cancer vaccine.
Sipuleucel-T (PROVENGE; Dendreon) is the first therapeutic cancer vaccine to be approved by the U.S. Food and Drug Administration. In men who have metastatic castration-resistant prostate cancer with no or minimal symptoms, sipuleucel-T prolongs median survival by 4.1 months compared with results in those treated with placebo. At 3 years, the proportion of patients in the vaccine group who were alive was 50% higher than that in the control group (31.7% versus 21.7%, respectively). Sipuleucel-T, which is designed to elicit an immune response to prostatic acid phosphatase, uses the patient's own immune system to recognize and combat his cancer. Currently, no other agents are available that offer a survival benefit for this population of asymptomatic patients who have not been treated with chemotherapy, except for docetaxel (whose inherent toxicities often lead patients and physicians to delay administration until symptoms develop). Straightforward strategies to increase the efficacy of sipuleucel-T are likely to provide even greater benefit. The preclinical and clinical development of sipuleucel-T is reviewed, and approaches to enhance efficacy are considered herein.


Cognitive aging and the hippocampus: how old rats represent new environments.
Spatial learning impairment in aged rats is associated with changes in hippocampal connectivity and plasticity. Several studies have explored the age-related deficit in spatial information processing by recording the location-specific activity of hippocampal neurons (place cells). However, these studies have generated disparate characterizations of place cells in aged rats as unstable (Barnes et al., 1997), resistant to change (Tanila et al., 1997b; Oler and Markus, 2000; Wilson et al., 2003), or delayed in using external cues (Rosenzweig et al., 2003). To reconcile these findings, we recorded place cells from aged and young rats as they repeatedly explored both a highly familiar environment and an initially novel environment, and we repeatedly tested whether the place fields formed in the novel environment were anchored by external cues. Initially, spatial representations in aged rats were abnormally maintained between the familiar and novel environments. Then, new representations were formed but were also delayed in becoming anchored to the external landmarks. Finally, even when the new spatial representations became bound to the landmarks, they were multi-stable across repetitive exposures to the formerly novel environment. These observations help to reconcile previously divergent characterizations of spatial representation in aged rats and suggest a model of cognitive aging and hippocampal function.


Myelosuppression of thrombocytes and monocytes is associated with a lack of synergy between chemotherapy and anti-VEGF treatment.
Chemotherapeutic agents that have shown improved patient outcome when combined with anti-vascular endothelial growth factor (VEGF) therapy were recently identified to induce the mobilization of proangiogenic Tie-2-expressing monocytes (TEMs) and endothelial progenitor cells (EPCs) by platelet release of stromal cell-derived factor 1α (SDF-1α). VEGF blockade was found to counteract cell mobilization. We aimed to determine why agents like gemcitabine do not elicit TEM and EPC recruitment and may therefore lack synergy with anti-VEGF therapy. Locally advanced pancreatic cancer patients (n = 20) were monitored during 16 weeks of neoadjuvant therapy. Treatment was based on gemcitabine with or without the addition of bevacizumab. Blood levels of proangiogenic cell populations and angiogenesis factors were determined in 2-week intervals. The lack of EPC mobilization during gemcitabine therapy was associated with severe thrombocytopenia and reduced SDF-1α blood concentrations. Furthermore, myelosuppression by gemcitabine correlated significantly with loss of TEMs. With respect to angiogenic factors stored and released by platelets, plasma levels of the angiogenesis inhibitor thrombospondin 1 (TSP-1) were selectively decreased and correlated significantly with thrombocytopenia in response to gemcitabine therapy. A thorough literature screen identified thrombocytopenia as a common feature of chemotherapeutic agents that lack synergy with anti-VEGF treatment. Our results on gemcitabine therapy indicate that myelosuppression (in particular, with respect to thrombocytes and monocytes) interferes with the mobilization of proangiogenic cell types targeted by bevacizumab and may further counteract antiangiogenic therapy by substantially reducing the angiogenesis inhibitor TSP-1.


Update on biliary and pancreatic sphincterotomy.
To summarize the indications, success rates and complications associated with endoscopic sphincterotomy and endoscopic balloon dilation (EBD). Pancreatic and/or biliary sphincterotomies are essential components of most current therapeutic endoscopic retrograde cholangiopancreatography (ERCP). A current large body of evidence has established biliary sphincterotomy as effective in extraction of bile duct stones. The most common complications of biliary sphincterotomy are post-ERCP pancreatitis, as well as acute or delayed hemorrhage, the risks for which can be stratified according to well described patient and procedure related factors. Evidence is accumulating that pancreatic sphincterotomy is useful in at least some settings for treatment of sphincter of Oddi dysfunction, chronic pancreatitis, and pancreas divisum. EBD provides an adjunct or an alternative to biliary sphincterotomy for extraction of stones from the bile duct when routine biliary sphincterotomy is inadequate or risk excessive. Sphincterotomy and EBD are useful in managing a variety of pancreatobiliary conditions. Attention to risks of these procedures is essential for their efficacy and safety.


Effects of molting on the visual acuity of the blue crab, Callinectes sapidus.
In crustaceans with compound eyes, the corneal lens of each facet is part of the exoskeleton and thus shed during molting. Here we used an optomotor assay to evaluate the impact of molting on visual acuity (as measured by the minimum resolvable angle, α(min)) in the female blue crab, Callinectes sapidus. We found that visual acuity decreases substantially in the days prior to molting and is gradually recovered after molting. Four days prior to molting, α(min) was 1.8 deg (N=5), a value approximating the best possible acuity in this species. In the 24 h before ecdysis occurred, α(min) increased to 15.0 deg (N=12), corresponding to an eightfold drop in visual acuity. Within 6 days after molting, α(min) returned to the pre-molting value. Micrographs of C. sapidus eyes showed that a gap between the corneal lens and the crystalline cone first appeared approximately 5 days prior to shedding and increased in width as the process progressed. This separation was likely responsible for the loss of visual acuity observed in behavioral tests. In blue crabs, mating is limited to the period of the female's pubertal molt, and a reduction in acuity during this time may have an effect on the sensory cues used in female mate choice. The results described here may be broadly applicable to all arthropods that molt and have particular importance for crustaceans that molt multiple times in their lifetime or have mating cycles that are paired with molting.


An Evaluation and Ranking of Children's Hospital Websites in the United States.
Children's hospitals are faced with the rising need for technological innovation. Their prospective health care consumers, who increasingly depend on the Web and social media for communication and consumer engagement, drive this need. As patients and family members navigate the Web presence of hospitals, it is important for these specialized organizations to present themselves and their services efficiently. The purpose of this study was to evaluate the website content of children's hospitals in order to identify opportunities to improve website design and create benchmarks to judge improvement. All websites associated with a children's hospital were identified using a census list of all children's hospitals in the United States. In March of 2014, each website and its social media were evaluated using a Web crawler that provided a 5-dimensional assessment that included website accessibility, marketing, content, technology, and usability. The 5-dimensional assessment was scored on a scale ranging from 0 to 10 with positive findings rated higher on the scale. Websites were ranked by individual dimensions as well as according to their average ranking across all dimensions. Mean scores of 153 websites ranged from 5.05 to 8.23 across all 5 dimensions. Results revealed that no website scored a perfect 10 on any dimension and that room exists for meaningful improvement. Study findings allow for the establishment of baseline benchmarks for tracking future website and social media improvements and display the need for enhanced Web-based consumer engagement for children's hospitals.


Men classified as hypo- or hyperresponders to dietary cholesterol feeding exhibit differences in lipoprotein metabolism.
The purpose of this study was to evaluate the differences that occur within the plasma compartment of normolipidemic men, classified on the basis of their response to prolonged consumption of additional dietary cholesterol. Using a crossover design, 40 men aged 18-57 y were randomly allocated to an egg (640 mg/d additional dietary cholesterol) or placebo group (0 mg/d additional dietary cholesterol), for two 30-d periods, which were separated by a 3-wk washout period. Subjects were classified as hypo- [increase in plasma total cholesterol (TC) of <0.05 mmol/L for each additional 100 mg of dietary cholesterol consumed] or hyperresponders (increase in TC of > or =0.06 mmol/L for each additional 100 mg of dietary cholesterol consumed) on the basis of their plasma reaction to the additional dietary cholesterol provided. Male hyporesponders did not experience an increase in LDL cholesterol (LDL-C) or HDL cholesterol (HDL-C) during the egg period, whereas both lipoproteins were significantly (P < 0.0001 and P < 0.05, respectively) elevated in hyperresponders. Although the LDL/HDL ratio was increased in male hyperresponders after the high cholesterol period, the mean increase experienced by this population was still within National Cholesterol Education Program guidelines. Furthermore, male hyperresponders had higher lecithin cholesterol acyltransferase (P < 0.05) and cholesteryl ester transfer protein (P < 0.05) activities during the egg period, which suggests an increase in reverse cholesterol transport. These data suggest that additional dietary cholesterol does not increase the risk of developing an atherogenic lipoprotein profile in healthy men, regardless of their response classification.


Neuroprotection afforded by targeting G protein-coupled receptors in heteromers and by heteromer-selective drugs.
G protein-coupled receptors (GPCRs) are the target of hundreds of approved drugs. Although these drugs were designed to target individual receptors, it is becoming increasingly apparent that GPCRs interact with each other to form heteromers. Approved drug targets are often part of a GPCR heteromer, and therefore new drugs can be developed with heteromers in mind. This review presents several strategies to selectively target GPCRs in heteromeric contexts, namely, taking advantage of i) heteromer-mediated biased agonism/signalling, ii) discovery of drugs with higher affinity for the receptor if it is part of a heteromer (heteromer selective drugs), iii) allosteric compounds directed against the interacting transmembrane domains and, eventually, iv) antagonists that block both GPCRs in a heteromer. Heteromers provide unique allosteric sites that should help designing a new type of drug that by definition would be a heteromer selective drug. The review also provides examples of rhodopsin-like class A receptors in heteromers that could be targeted to neuroprotect and/or delay the progression of diseases such as Parkinson's and Alzheimer's. GPCRs in heteromers (GriH) with the potential to address dyskinesias, a common complication of dopaminergic replacement therapy in parkinsonian patients, are also described.


The enzymatic activity of fungal xylanase is not necessary for its elicitor activity.
Fungal xylanases from Trichoderma spp. are potent elicitors of defense responses in various plants. To determine whether enzymatic activity is necessary for elicitor activity, we used site-directed mutagenesis to reduce the catalytic activity of xylanase II from Trichoderma reesei. For this, the glutamic acid residue at position 210, which is part of the active center in this family of enzymes, was changed to either aspartic acid (E210D) or serine (E210S). Wild-type and mutated forms of xylanase II were expressed in yeast cells and purified to homogeneity. Compared with the wild-type form of xylanase II, E210D had >100-fold and E210S 1,000-fold lower enzymatic activity. In contrast, these mutated forms showed no comparable drop in elicitor activity. They fully stimulated medium alkalinization and ethylene biosynthesis in suspension-cultured tomato (Lycopersicon esculentum) cells, as well as hypersensitive necrosis in leaves of tomato and tobacco (Nicotiana tabacum) plants. These results provide direct evidence that enzyme activity is not necessary for elicitor activity of fungal xylanase.


Affinity-based microarrays for proteomic analysis of cancer tissues.
Based on about a decade of technical developments in analysing the human proteome with antibody microarrays and experience in performing such analyses, now there are the means at hand for detailed and simultaneously global investigations of this kind. Many technical aspects have been dealt with of both the microarray format itself - such as overcoming kinetic and mass transport limitations and thus achieving accurate measurements - and ancillary processes - such as extraction procedures that provide good protein solubilisation, produce reproducible yields and preserve the native protein conformation as much as possible. The overall analysis process is robust and reproducible, highly sensitive down to the level of single-molecule detection and permits an analysis of several parameters on many molecules at a time. While the study of body liquids is widely applied, analyses of tissue proteomes are still scarce. However, conditions do exist to perform the latter at a quality level that meets the standards for clinical applications. This review highlights methodological aspects relevant for a biomedically useful analysis of cellular samples and discusses the potential of such studies, in particular, in view of personalised medicine approaches.


Groin pain associated with ultrasound finding of inguinal canal posterior wall deficiency in Australian Rules footballers.
To investigate the prevalence of inguinal canal posterior wall deficiency (sports hernia) in professional Australian Rules footballers using an ultrasound technique and correlate the results with the clinical symptom of groin pain. Thirty five professional Australian footballers with and without groin pain were investigated blind with a dynamic high resolution ultrasound technique for presence of posterior wall deficiency. Fourteen players had a history of significant recent groin pain and ten of these were found to have bilateral inguinal canal posterior wall deficiency (p < 0.01). The relative risk for a history of groin pain with bilateral deficiency was 8.0 (95% confidence interval 1.73 to 37.1). Groin pain was also found to be associated with increasing age (p < 0.01) which was an independent risk factor. Surgical, clinical, and ultrasound follow up for players who underwent hernia repair confirmed the validity of ultrasound as a diagnostic tool. Dynamic ultrasound examination is able to detect inguinal canal posterior wall deficiency in young males with no clinical signs of hernia. This condition is very prevalent in professional Australian Rules footballers, including some who are asymptomatic. There was a correlation between bilateral deficiency and groin pain, although the temporal relationship between the clinical and ultrasound findings is not established by the current study. Ultrasound shows promise as a diagnostic tool in athletes with chronic groin pain who are considered possible candidates for hernia repair.


Phase-slope and group-dispersion calculations in the frequency domain by simple optical low-coherence reflectometry.
We report a new method by which phase slope and group dispersion can be calculated with a simple optical low-coherence reflectometer to quantify physiological conditions. A discrete-time signal processing algorithm based on the first and second derivatives of the phase with respect to wave number was developed from discrete-time Fourier properties. The algorithm avoids the 2pi ambiguity associated with most phase unwrapping. Experimental data collected by use of well-characterized optical materials validated the algorithm, which was minimally sensitive to phase noise. The group dispersion of bovine cornea was measured at various hydrations and was significantly dependent on hydration. The results suggest that group dispersion is an indicator of corneal alterations.


Seasonal major affective disorder.
A seasonal affective disorder characterized by mild affective symptoms has recently been described. The author presents four cases of severe major affective disorder with a seasonal pattern and discusses the clinical relevance of seasonality.


One-year safety and effectiveness of the Agent paclitaxel-coated balloon for the treatment of small vessel disease and in-stent restenosis.
The Agent device consists of a semi-compliant balloon catheter, which is coated with a therapeutic low-dose formulation of paclitaxel (2 µg/mm2) blended with an inactive excipient acetyl-tri-n-butyl citrate (ATBC). AGENT Japan SV is a randomized controlled study that enrolled 150 patients from 14 Japanese sites treated with Agent or SeQuent Please paclitaxel-coated balloon. This study also includes a single-arm substudy evaluating the safety and effectiveness of Agent in patients with in-stent restenosis (ISR). Patients with a single de novo native lesion (lesion length ≤ 28 mm and reference diameter ≥ 2.00 to < 3.00 mm) were randomized 2:1 to receive either Agent (n = 101) or SeQuent Please (n = 49). The ISR substudy enrolled 30 patients with lesion length ≤ 28 mm and reference diameter ≥ 2.00 to ≤ 4.00 mm. In the SV RCT, target lesion failure (TLF) at 1 year occurred in four patients treated with Agent (4.0%) versus one patient with SeQuent Please (2.0%; P = 1.00). None of the patients in either treatment arm died. There were no significant differences in the rates of myocardial infarction, target lesion revascularization and target lesion thrombosis through 1 year. In the ISR substudy, the 1-year rates of TLF and target lesion thrombosis were 6.7% and 0.0%, respectively. These data support the safety and effectiveness of the Agent paclitaxel-coated balloon in patients with small vessels and ISR.


Continuous low-dose therapy with vinblastine and VEGF receptor-2 antibody induces sustained tumor regression without overt toxicity.
Various conventional chemotherapeutic drugs can block angiogenesis or even kill activated, dividing endothelial cells. Such effects may contribute to the antitumor efficacy of chemotherapy in vivo and may delay or prevent the acquisition of drug-resistance by cancer cells. We have implemented a treatment regimen that augments the potential antivascular effects of chemotherapy, that is devoid of obvious toxic side effects, and that obstructs the development of drug resistance by tumor cells. Xenografts of 2 independent neuroblastoma cell lines were subjected to either continuous treatment with low doses of vinblastine, a monoclonal neutralizing antibody (DC101) targeting the flk-1/KDR (type 2) receptor for VEGF, or both agents together. The rationale for this combination was that any antivascular effects of the low-dose chemotherapy would be selectively enhanced in cells of newly formed vessels when survival signals mediated by VEGF are blocked. Both DC101 and low-dose vinblastine treatment individually resulted in significant but transient xenograft regression, diminished tumor vascularity, and direct inhibition of angiogenesis. Remarkably, the combination therapy resulted in full and sustained regressions of large established tumors, without an ensuing increase in host toxicity or any signs of acquired drug resistance during the course of treatment, which lasted for >6 months. This article may have been published online in advance of the print edition. The date of publication is available from the JCI website, http://www.jci.org.


[Reflection microscopy of interchromosomal filaments].
After Giemsa staining of metaphases, reflection analysis reveals interchromosomal filaments. Either simple or in bundles, these filaments connect two chromosomes or sometimes several of them in a chain-like fashion. The anchoring points are most often localized in telomeric regions (64%). The form and the location of the chromosomes can be modified by the traction forces at the level of the anchoring sites. For those located outside the telomeric regions, the distribution of the sites shows a good correlation with the size of the chromosomal arms, except for the group D which is overrepresented.


Vitamin D receptor genotypes and intestinal calcium absorption in postmenopausal women.
Several studies have shown that bone mass and bone turnover are genetically determined. This genetic component is thought to be mediated in part by polymorphisms at the vitamin D receptor (VDR) locus, even though the underlying molecular mechanisms are still unknown. To evaluate a possible site of differential action of the VDR gene alleles we examined their correlation with intestinal calcium absorption in 120 Caucasian postmenopausal women (aged 61 +/- 0.6 years). VDR gene polymorphisms for Apa I, Bsm I, and Taq I restriction endonucleases were assessed by Southern blotting analysis. The most common genotypes observed in our population were AaBbTt (37%), AABBtt (20%), aabbTT (15%), AabbTT (15%), and AABbTt (9%). Although there was some evidence of 13% higher lumbar BMD values in aabbTT genotype with respect to AABBtt genotype, this difference of approximately 0.1 g/cm2 did not reach statistical significance, possibly because of the limited number of observations. On the contrary, no relationship was found between genotypes and femoral neck BMD values. Intestinal calcium absorption was significantly lower in BB and tt genotypes than, in bb and TT genotypes, respectively, and in AABBtt genotype than in either aabbTT or AaBbTt genotypes (P = 0.0015 ANOVA). No significant differences in intact PTH, alkaline phosphatase, 25OHD3, and 1, 25(OH)2D3 were found among subjects with different VDR genotypes. These results are consistent with a possible role of VDR alleles on intestinal calcium absorption.


What does body size mean, from the "plant's eye view"?
Alternative metrics exist for representing variation in plant body size, but the vast majority of previous research for herbaceous plants has focused on dry mass. Dry mass provides a reasonably accurate and easily measured estimate for comparing relative capacity to convert solar energy into stored carbon. However, from a "plant's eye view", its experience of its local biotic environment of immediate neighbors (especially when crowded) may be more accurately represented by measures of "space occupancy" (S-O) recorded in situ-rather than dry mass measured after storage in a drying oven. This study investigated relationships between dry mass and alternative metrics of S-O body size for resident plants sampled from natural populations of herbaceous species found in Eastern Ontario. Plant height, maximum lateral canopy extent, and estimated canopy area and volume were recorded in situ (in the field)-and both fresh and dry mass were recorded in the laboratory-for 138 species ranging widely in body size and for 20 plants ranging widely in body size within each of 10 focal species. Dry mass and fresh mass were highly correlated (r2 > .95) and isometric, suggesting that for some studies, between-species (or between-plant) variation in water content may be unimportant and fresh mass can therefore substitute for dry mass. However, several relationships between dry mass and other S-O body size metrics showed allometry-that is, plants with smaller S-O body size had disproportionately less dry mass. In other words, they have higher "body mass density" (BMD) - more dry mass per unit S-O body size. These results have practical importance for experimental design and methodology as well as implications for the interpretation of "reproductive economy"-the capacity to produce offspring at small body sizes-because fecundity and dry mass (produced in the same growing season) typically have a positive, isometric relationship. Accordingly, the allometry between dry mass and S-O body size reported here suggests that plants with smaller S-O body size-because of higher BMD-may produce fewer offspring, but less than proportionately so; in other words, they may produce more offspring per unit of body size space occupancy.


Application of the International Classification of Functioning, Disability and Health in children with neurofibromatosis type 1: a review.
The World Health Organization's International Classification of Functioning, Disability and Health adapted for children and young people (ICF-CY) is a framework for describing and classifying health and health-related states. The aim of the present study was to review literature on neurofibromatosis type 1 (NF1) using ICF-CY guidelines and to highlight findings about the quality of life of children with NF1. Electronic databases were searched to identify studies involving children with NF1. Eligible studies were classified according to ICF-CY categories. Children with NF1 have a variety of cognitive and other deficits. However, very little information is available on the impact of these deficits on their daily life. Despite the broad range of functional and structural deficits in children with NF1, the functional assessment of these children remains largely unexplored. Future studies should aim at evaluating the participation of children with NF1 in various situations, using tools with high real-world validity.


Multifunctional magnetic-optical nanoparticle probes for simultaneous detection, separation, and thermal ablation of multiple pathogens.
Multifunctional nanoparticles possessing magnetization and near-infrared (NIR) absorption have warranted interest due to their significant applications in magnetic resonance imaging, diagnosis, bioseparation, target delivery, and NIR photothermal ablation. Herein, the site-selective assembly of magnetic nanoparticles onto the ends or ends and sides of gold nanorods with different aspect ratios (ARs) to create multifunctional nanorods decorated with varying numbers of magnetic particles is described for the first time. The resulting hybrid nanoparticles are designated as Fe(3)O(4)-Au(rod)-Fe(3)O(4) nanodumbbells and Fe(3)O(4)-Au(rod) necklacelike constructs with tunable optical and magnetic properties, respectively. These hybrid nanomaterials can be used for multiplex detection and separation because of their tunable magnetic and plasmonic functionality. More specifically, Fe(3)O(4)-Au(rod) necklacelike probes of different ARs are utilized for simultaneous optical detection based on their plasmon properties, magnetic separation, and photokilling of multiple pathogens from a single sample at one time. The combined functionalities of the synthesized probes will open up many exciting opportunities in dual imaging for targeted delivery and photothermal therapy.


Synthesis, ADMET Properties, and Biological Evaluation of Benzothiazole Compounds Targeting Chemokine Receptor 2 (CXCR2).
Herein we describe the synthesis and biological evaluation of a series of novel benzothiazoles based on a diaryl urea scaffold previously reported in some allosteric chemokine receptor 2 (CXCR2) inhibitors. From a library of 41 new compounds, 17 showed significant inhibition of CXCR2, with IC50 values less than 10 μm and selectivity over CXCR4. Our ADMET simulations suggest favorable drug-like properties for the active compounds. Importantly, we developed a predictive model that can distinguish active from inactive compounds; this will serve as a valuable tool to guide the design of optimized compounds to be evaluated in preclinical models.


Pathological stage does not alter the prognosis for renal lesions determined to be stage T1 by computerized tomography.
Pathological stage has been the most widely used prognosticator for evaluating surgically managed cases of renal cell carcinoma. Minimally invasive surgical approaches are being increasingly used to treat small masses for which traditionally pathological information is lacking (morcellation) or absent (radio frequency ablation or cryoablation). Preoperative cross-sectional imaging by computerized tomography (CT) or magnetic resonance imaging has been used to stage renal tumors clinically but it can lead to variances with traditional pathological staging systems, particularly with respect to microscopic invasion beyond the renal capsule. In this study we assessed whether radiographically staged clinical T1 lesions that were pathological T1 behave differently than those that were clinical stage T1 and up staged to pT3a. The records of 296 patients who underwent surgical treatment for renal cell carcinoma at The Johns Hopkins Hospital between 1990 and 1999 were retrospectively reviewed. All patients had undergone preoperative CT or magnetic resonance imaging, which was used to assign a clinical stage and size (largest diameter) to each tumor in accordance with the 1997 TNM staging system. Following surgical resection pathological stage, size and tumor grade were determined. Only the 186 patients with clinical T1 tumors were included in this analysis. Of the 186 patients who were clinically found to have T1 lesions 125 (67%) had pathological T1 and 57 (31%) had pathological T3a lesions. All surgical margins and lymph nodes were negative at surgical resection. Mean tumor size +/- SD was 3.9 +/- 1.5 cm for pT1 lesions and 3.8 +/- 1.5 cm for pT3a lesions. When comparing these pathological groups using Kaplan-Meier analysis, 5-year recurrence-free survival was not statistically different in patients with pT1 and pT3a lesions (90.6 and 97.5%, respectively). Patients in whom the initial classification of T1 renal cell carcinoma by CT was up graded to T3a on pathological analysis (invasion of fat within Gerota's fascia) showed the same recurrence-free survival rate as patients with pathologically confirmed T1 lesions. Thus, smaller tumors (less than 7 cm) that are up graded to T3a based on capsule invasion behave much like T1 tumors and exact pathological T staging does not appear to impact overall survival.


Determinants of neural tube defects among women who gave birth in hospitals in Eastern Ethiopia: evidence from a matched case control study.
Neural tube defects (NTDs) are severe birth defects caused by nutritional, genetic or environmental factors. Because NTDs continue to have a significant health and economic impact on children and community at large, it is crucial to investigate potential risk factors in order to develop novel approaches to NTDs prevention. Determinants for the development of NTDs differ by country, region as well as within the country. The objective of this study was to identify the determinants of NTDs among newborns delivered in three hospitals found in eastern Ethiopia. A hospital-based matched case-control study was conducted among 138 cases and 138 control women who delivered in three teaching hospitals in Eastern Ethiopia in 2021. Data were collected using a structured and pre-tested interviewer-administered questionnaire. Cases were mothers who delivered a neonate with any type of NTDs regardless of gestational age or fetal viability, whereas controls were mothers who delivered an apparently healthy newborn. Chi-square was used to assess the significant difference between the two groups. Conditional logistic regression model was used to generate adjusted odds ratio with its corresponding 95% confidence intervals and compare the two groups. Anencephaly (51.4%) and spinal bifida (34.1%) were the most frequently observed NTDs. None of study participants took preconception folic acid supplementation. Being a non-formal mothers (AOR = 0.34, 95% CI: 0.12-0.92, P = 0.034), rural residence, (AOR = 3.4, 95% CI: 1.18-9.78, P = 0.023), history of spontaneous abortion (AOR = 2.95, 95% CI: 1.15-7.55, P = 0.023), having severe anemia (AOR = 3.4, 95% CI: 1.17-9.87, P = 0.024), history of fever or cold (AOR = 2.75; 95% CI: 1.05-7.15, P = 0.038), and an exposure to various agro-chemicals (AOR = 3.39, 95% CI: 1.11-10.3, P = 0.032) were independent determinants of NTDs. In this study, NTDs were associated to several determinant factors in the area, including residential area, history of spontaneous abortion, severe anemia, fever/cold, antibiotic use before or during early pregnancy, and exposure to agrochemicals. Addressing the identified determinants is critical in averting the incidence of NTDs in the study area. Moreover, more research is needed to investigate women's dietary practices as well as the practice of preconception folic acid supplementation for pregnant women in Ethiopia's current health care system.


[Determination by ultrasound of chorionicity in twin pregnancy].
Twins, specially those of monochorionic and monoamniotic pregnancies, are exposed to many perinatal risks and complications. The objective of this study was to evaluate the usefulness of ultrasonographic determination of the chorionicity of human placental by counting the number of layers in amniotic membranes. Thirty-eight patients with twin pregnancies were studied prospectively. The ultrasonographic evaluation of membranes layers was made only once, between the weeks and 16 and 30 of pregnancy. When two layers were identified, the placentation was determined as monochorionic, and when four layers were seen, the diagnosis of bichorionic placentation was made. The type of chorionicity was confirmed by histologic study of the placenta. With ultrasound, the chorionicity was correctly determined in 36 out of 38 cases, for a total predictive value of 94.6%. The capacity for diagnosing bichorionic (4 layers) placentation was 100% (22/22) and 87.6% (14/16) for monochorionic (2 layers) placentation. The ultrasonographic evaluation of the amniotic membranes number is an efficient method to recognize the chorionicity of placenta and constitutes an useful and simple method giving important information in perinatal prognosis.


Some pathogenic features of viral hepatitis.
Some pathogenic features of viral hepatitis (VH) are analysed on the ground of literature data and personal results. Correlations concerning blood group distribution in the case of apparently asymptomatic HBsAg carriage and of acute and chronic VH, as well as correlations with individual characteristics such as age and sex a.o., are presented. The possibility that other genetic and epigenetic factors may help to a better knowledge of VH pathogeny is discussed.


Mechanism of cellular oxidation stress induced by asymmetric dimethylarginine.
The mechanism by which asymmetric dimethylarginine (ADMA) induces vascular oxidative stress is not well understood. In this study, we utilized human umbilical vein endothelial cells (HUVEC) to examine the roles of ADMA cellular transport and the uncoupling of endothelial nitric oxide synthase (eNOS) in contributing to this phenomenon. Dihydroethidium (DHE) fluorescence was used as an index of oxidative stress. Whole cells and their isolated membrane fractions exhibited measureable increased DHE fluorescence at ADMA concentrations greater than 10 μM. ADMA-induced DHE fluorescence was inhibited by co-incubation with L-lysine, tetrahydrobiopterin (BH(4)), or L-nitroarginine methyl ester (L-NAME). Oxidative stress induced in these cells by angiotensin II (Ang II) were unaffected by the same concentrations of L-lysine, L-NAME and BH(4). ADMA-induced reduction in cellular nitrite or nitrite/nitrate production was reversed in the presence of increasing concentrations of BH(4). These results suggest that ADMA-induced DHE fluorescence involves the participation of both the cationic transport system in the cellular membrane and eNOS instead of the Ang II-NADPH oxidase pathway.


Estrogen protection against coronary heart disease: are the relevant effects of estrogen mediated through its effects on uterus--such as the induction of menstruation, increased bleeding, and the facilitation of pregnancy?
Women in the age group of 20-50 are shown to have much less susceptibility to Coronary Heart Disease (CHD) and other atherosclerotic diseases as compared to men. Exact cause of which is not precisely known and estrogen is constantly shown to be associated with this phenomenon. Improvement of serum HDL concentration and improvement of endothelial functions are some of the proposed mechanisms through which estrogen is believed to mediate this effect. Estrogen therapy however has failed to protect women with bilateral oophorectomy and hysterectomy. Similarly inability of endogenous estrogen to protect women, who have undergone hysterectomy with functioning ovaries from CHD, questions the currently accepted mechanisms through which estrogen brings about these protective effects. Ineffectiveness of estrogen therapy as prophylaxis against CHD in men further questions the credibility of the currently accepted protective mechanisms of estrogen. Estrogen has variety of effects of on uterus, to induce menstruation, to induce bleeding, facilitative role in pregnancy, fetal growth and development. As these physiological effects either directly or indirectly result in loss of cholesterol from cardiovascular compartment, it is proposed that cholesterol-losing effects of estrogen are more important than its presently believed beneficial effects. The small amount that is lost causes movement of cholesterol from atheroma towards plasma and thereby retards the progress of atherosclerosis. These cholesterol-losing effects of estrogen enable women to enjoy freedom from CHD during their reproductive age, as compared to men of comparable age group. Statistical data obtained from blood donors indirectly support the proposed hypothesis.


Sequence analysis of the Chlamydomonas reinhardtii flagellar alpha dynein gene.
Flagellar outer row dynein ATPases have been used extensively as model systems for studies of microtubule-based motility. Previously full-length sequences were only available for two of the three catalytic heavy-chain subunits (DHCs) of this enzyme. We have completed the sequence of an 18-kb genomic region encoding the Chlamydomonas reinhardtii flagellar outer row dynein alpha heavy chain. Unlike the beta- and gamma-subunits, DHC alpha is not required for assembly of other outer row dynein proteins, except for a tightly associated light chain, and thus occupies a unique position within this enzyme complex. The predicted 4,499 residue protein retains sequence homology to other dynein heavy chains throughout its central and C-terminal regions but lacks homology to any other dyneins in the first 1,000 amino acids, which may account for its unusual assembly properties. This N-terminal domain of DHC alpha contains a repetitive sequence rich in alanines, prolines, and glutamic acids. Within the more homologous C-terminal region, which includes the catalytic domain, three short sequences unique to DHC alpha may account for its specific catalytic properties and in vivo phosphorylation pattern.


Effects of Localized Cold Therapy on Pain in Postoperative Spinal Fusion Patients: A Randomized Control Trial.
Cold therapy used in the sports medicine settings has been found to be effective in reducing postoperative pain; however, there are limited studies that examine the effect of cold therapy on postoperative pain in patients with posterior lumbar spinal fusion. The purpose of this study was to determine the effects of cold on postoperative spine pain and add to the body of knowledge specific to practical application of cold therapy in the spine surgery setting. Researchers used a two-group randomized control design to evaluate the effects of local cold therapy on postoperative pain and analgesia use after lumbar spinal fusion surgery. The primary outcome was postoperative pain. Secondary outcomes included analgesia use and perceived benefit of cold therapy. The intervention (cold) group had a marginally greater reduction in mean Numerical Rating Scale score across all 12 pain checks (M ± SD = -1.1 ± 0.8 points reduction vs. -1.0 ± 0.8 points reduction, p = .589). On average, the intervention group used less morphine equivalents (M ± SD = 12.6 ± 31.5 vs. 23.7 ± 40.0) than the control group across pain checks seven to 12 (p = .042). This study provides additional evidence to support the use of cold therapy as an adjuvant pain management strategy to optimize pain control and reduce opioid consumption following spine fusion surgical procedures.


Electric field controlled transport of water in graphene nano-channels.
Motivated by electrowetting-based flow control in nano-systems, water transport in graphene nano-channels is investigated as a function of the applied electric field. Molecular dynamics simulations are performed for deionized water confined in graphene nano-channels subjected to opposing surface charges, creating an electric field across the channel. Water molecules respond to the electric field by reorientation of their dipoles. Oxygen and hydrogen atoms in water face the anode and cathode, respectively, and hydrogen atoms get closer to the cathode compared to the oxygen atoms near the anode. These effects create asymmetric density distributions that increase with the applied electric field. Force-driven water flows under electric fields exhibit asymmetric velocity profiles and unequal slip lengths. Apparent viscosity of water increases and the slip length decreases with increased electric field, reducing the flow rate. Increasing the electric field above a threshold value freezes water at room temperature.


Vulnerability and security in seriously ill patients in intensive care.
The purpose of this research was to gain an understanding of the experience of being a seriously ill patient in an intensive care unit (ICU). Fourteen former patients, aged 17-71 years old, who had been in ICU 3-53 days, participated in focus group interviews 3-6 months after discharge. The focus groups met 3 times each for 1.5 hours, resulting in 13-14 hours of audiotaped discussions. The transcribed data were qualitatively analysed to identify themes representing participants' experiences. Vulnerability emerged as a central concept that captured the identified themes. The data reveal that patient vulnerability while in ICU was related to extreme physical and emotional dependency. Lack of information and depersonalizing care were associated with fear, anxiety and increased vulnerability. Lack of sleep and rest also contributed to patient fear and anxiety. Vulnerability decreased when patients were kept informed of what was occurring while in ICU, received care that was personalized to their individual needs, and when their families were present. The results of this study suggest that ICU patients' vulnerability may be decreased by the security that they experience when they are adequately informed about what is happening, and when nursing and medical care is personalized to their individual needs.


Liposome division by a simple bacterial division machinery.
We previously reconstituted Z rings in tubular multilamellar liposomes with FtsZ-YFP-mts, where mts is a membrane-targeting amphiphilic helix. These reconstituted Z rings generated a constriction force but did not divide the thick-walled liposomes. Here we developed a unique system to observe Z rings in unilamellar liposomes. FtsZ-YFP-mts incorporated inside large, unilamellar liposomes formed patches that produced concave distortions when viewed at the equator of the liposome. When viewed en face at the top of the liposome, many of the patches were seen to be small Z rings, which still maintained the concave depressions. We also succeeded in reconstituting the more natural, two-protein system, with FtsA and FtsZ-YFP (having the FtsA-binding peptide instead of the mts). Unilamellar liposomes incorporating FtsA and FtsZ-YFP showed a variety of distributions, including foci and linear arrays. A small fraction of liposomes had obvious Z rings. These Z rings could constrict the liposomes and in some cases appeared to complete the division, leaving a clear septum between the two daughter liposomes. Because complete liposome divisions were not seen with FtsZ-mts, FtsA may be critical for the final membrane scission event. We demonstrate that reconstituted cell division machinery apparently divides the liposome in vitro.


Indicators as a tool for the evaluation of effective national implementation of the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).
This paper briefly outlines the United Nations Globally Harmonized System of Classification and Labelling of Chemicals (GHS) and what is meant by GHS implementation in terms of policy-related information for decision-makers plus a framework of actions to enable such policies to be effectively undertaken. The paper discusses the need for simple indicators to measure progress with GHS implementation. Two groups of indicators have been described that comprehensively reflect basic components of GHS implementation. The first group was presented as aggregated indicators or indices, for national capability evaluation. Indicators to evaluate management performance linked to stated objectives were presented as the second group. Information from an Organisation for Economic Co-operation and Development (OECD) published questionnaire on GHS implementation and a questionnaire by the Intergovernmental Forum on Chemical Safety (IFCS) on chemical safety that included GHS have provided the basic information for applying the management capability indicators approach. Through the use of information in these two questionnaires, country-relevant and regional progress with GHS implementation was illustrated. Despite the GHS implementation target of 2008 set by the Johannesburg Plan of Implementation, very few countries have reported they have achieved this target. The simple management indicators proposed in this paper are based on adoption of a questionnaire and a numerical scoring system that could be used to chart progress towards GHS implementation. It was concluded, therefore, that the use of indicators as described in this paper does provide a quick survey of the overall situation in a country and facilitates comparison between countries across regions in terms of GHS implementation. Furthermore such results may indicate which countries require additional assistance with GHS implementation thus strengthening policies and actions across all sectors and involving governments, business, trade, and civil society.


mTOR modulates resistance to gemcitabine in lung cancer in an MTORC2 dependent mechanism.
Lung cancer has a poor prognosis partly due to a lack of response to treatments such as the chemotherapy drug gemcitabine. Combinations of chemotherapy drugs with signal transduction inhibitors may be more effective treatments. In this study we have investigated the impact of targeting the mTOR signalling pathway on the efficacy of gemcitabine in different cancer cell lines. Time-lapse microscopy, immuno-staining, and western blot techniques were used to evaluate the efficacy of applied treatments either in measuring phosphorylation levels of mTOR down-stream targets or in tracking down the fate of targeted cells. Reactive oxygen species and relative levels of protein phosphorylation were also quantified. For comparison between treated groups t-test and analysis of variance test were applied. Our data showed that mTORC1 has no role in sensitising A549 lung cancer cells to gemcitabine. However, targeting mTORC1/2 with the pharmacological inhibitor torin1 or by over-expressing Deptor, the negative regulator of mTOR signalling, sensitised these cells to gemcitabine. Silencing mTORC2, but not mTORC1, induced apoptosis and significantly improved the apoptosis-inducing effects of gemcitabine. Results also suggest that Rictor is required to maintain cell survival through modulating p38α, ERK1/2, RSK1/2/3 and the transcription factor STAT3. Multiple cell line comparisons revealed that PANC-1 pancreatic cancer cells were also sensitive to mTOR inhibition, but MCF7 breast cancer, MCF10A breast epithelial and H727 lung cancer cell lines were more resistant to the treatment. Inhibition of mTORC2 may have benefits in the treatment of gemcitabine resistant cancers, and the genetic background of the cell line may determine its response to mTOR inhibition.


Broncho-vaxom alleviates persistent allergic rhinitis in patients by improving Th1/Th2 cytokine balance of nasal mucosa.
Probiotics are mainly distributed in the mucosal system and have the ability to enhance mucosal barrier function and regulate immune responses. Broncho-Vaxom (BV), as a probiotic, has been applied to patients suffering from respiratory tract infections, but its potential effectiveness in allergic rhinitis (AR) has not been evaluated in human. This study aimed to investigate the clinical efficacy of BV in patients with persistent AR and to elucidate the underlying cellular mechanisms. Sixty patients with AR were enrolled to this study and were randomly assigned to the BV group (n=30) and the placebo group (n=30). Changes of clinical symptoms and laboratory parameters of allergic inflammation were measured at baseline visit, immediately after BV treatment, four weeks, and eight weeks after the BV treatment. After BV treatment, medication score in the BV group was significantly decreased compared with placebo group, along with a significant drop of the total nasal symptom score and the individual nasal symptom scores (itching score: 23.72±5.32%; nasal rhinorrhea score: 18.59±4.83%; sneezing score: 23.08±4.98%). The levels of IL-4 and IL-13 in nasal lavage were diminished remarkably while the level of INF-γ was markedly increased in the BV group. This rendered a significant reduction of the ratio of IL-4/INF-γ. Moreover, a decrease of eosinophils in nasal smear was observed after BV treatment. The BV-induced favorable changes sustained for at least four to eight weeks post BV treatment. Oral administration of BV offers remarkable and sustained efficacy in alleviating AR symptoms and may be considered as an alternative therapeutic strategy for patients with persistent AR. BV acts by improving the overall mucosal immunity via restoring and maintaining the normal Th1/Th2 cytokine balance as an underlying cellular/signaling mechanism.


Physical Activity, Weight, and Outcomes in Patients Receiving Chemotherapy for Metastatic Breast Cancer (C40502/Alliance).
Obesity and inactivity are associated with increased risk of cancer-related and overall mortality in breast cancer, but there are few data in metastatic disease. Cancer and Leukemia Group B 40502 was a randomized trial of first-line taxane-based chemotherapy for patients with metastatic breast cancer. Height and weight were collected at enrollment. After 299 patients enrolled, the study was amended to assess recreational physical activity (PA) at enrollment using the Nurses' Health Study Exercise Questionnaire. Associations with progression-free survival (PFS) and overall survival (OS) were evaluated using stratified Cox modeling (strata included hormone receptor status, prior taxane, bevacizumab use, and treatment arm). All statistical tests were 2-sided. A total of 799 patients were enrolled, and at the time of data lock, median follow-up was 60 months. At enrollment, median age was 56.7 years, 73.1% of participants had hormone receptor-positive cancers, 42.6% had obesity, and 47.6% engaged in less than 3 metabolic equivalents of task (MET) hours of PA per week (<1 hour of moderate PA). Neither baseline body mass index nor PA was statistically significantly associated with PFS or OS, although there was a marginally statistically significant increase in PFS (hazard ratio = 0.83, 95% confidence interval = 0.79 to 1.02; P = .08) and OS (hazard ratio = 0.81, 95% confidence interval = 0.65 to 1.02; P = .07) in patients who reported PA greater than 9 MET hours per week vs 0-9 MET hours per week. In a trial of first-line chemotherapy for metastatic breast cancer, rates of obesity and inactivity were high. There was no statistically significant relationship between body mass index and outcomes. More information is needed regarding the relationship between PA and outcomes.


Polymorphisms in xenobiotic metabolizing genes, intakes of heterocyclic amines and red meat, and postmenopausal breast cancer.
Heterocyclic amines (HCAs) are mutagenic compounds generated when meats are cooked at high temperature and for long duration. The findings from previous studies on the relation between HCAs and breast cancer are inconsistent, possibly because of genetic variations in the enzymes metabolizing HCAs. To evaluate whether the associations of intakes of estimated HCAs, meat-derived mutagenicity (MDM), and red meat with risk of postmenopausal breast cancer were modified by N-acetyltransferase 2 (NAT2) acetylator genotype or cytochrome P450 1A2-164 A/C (CYP1A2) polymorphism, we conducted a nested case-control study with 579 cases and 981 controls within a prospective cohort, the Nurses' Health Study. HCAs and MDM intakes were derived using a cooking method questionnaire administered in 1996. NAT2acetylator genotype, the CYP1A2 polymorphism, and intakes of HCAs, MDM, and red meat were not associated with risk of postmenopausal breast cancer. There was also no interaction between NAT2 acetylator genotype or CYP1A2 polymorphism and HCAs and MDM and red meat intake in relation to breast cancer. These results do not support the hypothesis that genetic polymorphisms of xenobiotic enzymes involved in the metabolism of HCAs may modify the associations between intakes of red meat or meat-related mutagens and breast cancer risk.


Influence of the COMT val(108/158)met polymorphism on continuous performance task indices.
The Continuous Performance Task (CPT) is a widely-used measure of sustained attention and impulsivity. Deficits in CPT performance have been found in several psychiatric disorders, such as Attention-Deficit/Hyperactivity Disorder (ADHD) and schizophrenia. Molecular genetic studies of CPT performance are currently limited and have generally revealed inconsistent findings. The current study tested the associations of the COMT val(108/158)met polymorphism with AX-CPT indices (i.e., omission and commission errors, d׳, and lnβ), as well as the variability of these indices across blocks, in a sample of clinic-referred and non-referred children (N=380). We found significant associations between COMT and variability in the Signal Detection Theory (SDT) indices d׳ and lnβ across blocks, as well as a statistical trend for association between COMT and commission errors. Higher externalizing psychopathology was associated with general impairment on AX-CPT performance, and for some indices (i.e., d׳ variability and lnβ variability) the effect of COMT was stronger at higher levels of psychopathology. Our findings support the role of COMT in components of CPT performance and highlight the potential utility of using SDT indices, particularly in relation to variability in performance. Moreover, our results suggest that for some indices the effect of COMT is stronger at higher levels of externalizing psychopathology. Our study yields some preliminary insights regarding the neurobiology of CPT performance, which may elucidate the mechanisms by which specific genes confer risk for various cognitive deficits, as well as relevant disorders characterized by these deficits.


Rewarding effects of AMPA administration into the supramammillary or posterior hypothalamic nuclei but not the ventral tegmental area.
We examined whether injections of the excitatory amino acid AMPA are rewarding when injected into the posterior hypothalamus and ventral tegmental area. Rats quickly learned to lever-press for infusions of AMPA into the supramammillary or posterior hypothalamic nuclei but failed to learn to lever-press for similar injections into the ventral tegmental areas. AMPA injections into the supramammillary nucleus, but not the ventral tegmental area, induced conditioned place preference. The rewarding effects of AMPA appear to be mediated by AMPA receptors, because coadministration of the AMPA antagonist CNQX blocked the rewarding effects of AMPA, and administration of the enantiomer R-AMPA did not mimic the rewarding effects. AMPA injections into the supramammillary nucleus, but not the ventral tegmental area, also increased extracellular dopamine concentrations in the nucleus accumbens. Pretreatment with the D1 dopamine antagonist SCH 23390 [R-(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine] led to extinction of AMPA self-administration. These findings implicate posterior hypothalamic regions in reward function and suggest that reward mechanisms localized around the ventral tegmental area are more complex than has been assumed recently.


Seriously depressed preschoolers.
In a prospective and comprehensive evaluation (including DSM-III diagnoses) of 1,000 preschoolers in a child development unit, nine children were found to meet the criteria for major depressive disorder. The symptoms of these children are described and compared with those of a control group of children from the same sample. The diagnosis of major depressive disorder was associated with physical symptoms, including somatic complaints, and the likelihood of parental abuse or neglect.


Does race affect outcomes in triple negative breast cancer?
There is discordance among studies assessing the impact of race on outcome of patients with Triple Negative Breast Cancer (TNBC). We assessed survival outcomes for African American (AA) versus Caucasian (CA) women with TNBC treated at an urban cancer center in Memphis, TN with a predominant AA patient population. Patients with Stage I-III TNBC were identified from our breast database. Event free survival (EFS) and Breast cancer specific survival (BCSS) were the primary outcome measures. Cox proportional hazards models were fitted for EFS and BCSS. Of the 124 patients, 71% were AA. No significant association between race and stage (P = 0.21) or menopausal status (P = 0.15) was observed. Median age at diagnosis was significantly lower for AA versus CA women (49.5 vs. 55 years, P = 0.024). 92% of the patients received standard neo/adjuvant chemotherapy, with no significant difference in duration and type of chemotherapy between the races. With a median follow up of 23 months, 28% of AA vs. 19% of CA women had an event (P = 0.37). 3 year EFS and BCSS trended favorably towards CA race (77% vs. 64%, log rank P = 0.20 and 92% vs. 76%, P = 0.13 respectively) with a similar trend noted on multiple variable modeling (EFS: HR 0.62, P = 0.29; BCSS: HR 0.36, P = 0.18). AA women >/=50 years at diagnosis had a significantly worse BCSS than the CA women in that age group (P = 0.012). Older AA women with TNBC have a significantly worse breast cancer specific survival than their CA counterparts. Overall, there is a trend towards lower survival for AA women compared to Caucasians despite uniformity of tumor phenotype and treatment. The high early event rate, irrespective of race, underscores the need for effective therapies for women with TNBC.