Does timing of axon outgrowth influence initial retinotectal topography in Xenopus?
The question of whether timing is involved in generating the topographic organization of the earliest embryonic projection from the retina to the tectum has been examined in Xenopus laevis. First, the normal schedule of axonal outgrowth from the retina to the tectum was characterized. Groups of axons originating from either dorsal or ventral extremes of the retina were labeled by in vitro incubation of sectors (one-quarter to one-third) of eye primordia in [3H]proline and their time courses of outgrowth were determined using light microscope autoradiography. Comparisons of the growth of dorsal and ventral nerve fiber populations showed that those from the dorsal retina leave the eye first, grow along the optic pathway, and reach the tectum roughly 6 hr ahead of those from the ventral retina. This stereotyped sequence of outgrowth schedules the development of the initial retinotectal map: first the ventrolateral tectum receives input from the dorsal retina (stage 37/38), and then the dorsomedial part receives input from ventral retina (stage 40). Second, to test whether the accurate timing of axon outgrowth and target invasion defines the spatial ordering of the earliest connections, the normal schedule of retinal fiber outgrowth was altered by substituting dorsal halves of young stage 21 eye primordia, labeled with [3H]proline, for those in older stage 27 embryos. These heterochronic transplants resulted in retarded outgrowth from the dorsal retina such that the original pioneer fibers reached the tectum at least 9 hr later than normal, arriving after ventral retinal fibers, thereby reversing the normal sequence of tectal invasion. Despite this, the initial pattern of tectal innervation remained spatially normal. It is concluded that the retinotectal map is generated not by the temporal sequence of retinal axon outgrowth but, rather, by selective means of neuronal recognition.


Developmental sequence of periventricular leukomalacia. Correlation of ultrasound, clinical, and nuclear magnetic resonance functions.
The evolution of severe periventricular leukomalacia was followed by ultrasonography in three newborn infants, and the subsequent myelination of the brain was assessed by nuclear magnetic resonance imaging. Four stages of periventricular leukomalacia could be identified by ultrasonography; (1) initial congestion, followed by (2) relative normalisation, (3) development of cysts, and (4) resolution of cysts but development of ventricular enlargement. All infants exhibited abnormal neurological signs from 36 weeks conceptual age and had unequivocal signs of cerebral palsy by 6 to 9 months of age. One infant became cortically blind but the other two seemed to have normal vision. Nuclear magnetic resonance imaging showed some abnormality of the ventricular system and delayed myelination in all three infants. The delay was most noticeable in the opticothalamic region, which was also the site of the most extensive lesions observed on ultrasonography. Progress in myelination was observed in the infants where a repeat scan was performed.


Prostate Cancer-Associated Kallikrein-Related Peptidase 4 Activates Matrix Metalloproteinase-1 and Thrombospondin-1.
Prostate cancer metastasis to bone is terminal; thus, novel therapies are required to prevent end-stage disease. Kallikrein-related peptidase 4 (KLK4) is a serine protease that is overproduced in localized prostate cancer and is abundant in prostate cancer bone metastases. In vitro, KLK4 induces tumor-promoting phenotypes; however, the underlying proteolytic mechanism is undefined. The protein topography and migration analysis platform (PROTOMAP) was used for high-depth identification of KLK4 substrates secreted by prostate cancer bone metastasis-derived PC-3 cells to delineate the mechanism of KLK4 action in advanced prostate cancer. Thirty-six putative novel substrates were determined from the PROTOMAP analysis. In addition, KLK4 cleaved the established substrate, urokinase-type plasminogen activator, thus validating the approach. KLK4 activated matrix metalloproteinase-1 (MMP1), a protease that promotes prostate tumor growth and metastasis. MMP1 was produced in the tumor compartment of prostate cancer bone metastases, highlighting its accessibility to KLK4 at this site. KLK4 further liberated an N-terminal product, with purported angiogenic activity, from thrombospondin-1 (TSP1) and cleaved TSP1 in an osteoblast-derived matrix. This is the most comprehensive analysis of the proteolytic action of KLK4 in an advanced prostate cancer model to date, highlighting KLK4 as a potential multifunctional regulator of prostate cancer progression.


Rural-Urban Differences in Dietary Behavior and Obesity: Results of the Riskesdas Study in 10-18-Year-Old Indonesian Children and Adolescents.
Obesity has become a significant problem for developing countries, including Indonesia. High duration of sedentary activity and high intake of unhealthy foods were associated with high risk of overweight and obesity. The objective of this study was to compare the distributions of sedentary activity and dietary behavior with overweight/obesity risks between urban and rural areas among children and adolescents aged 10-18 years in Indonesia. This is a cross-sectional study. Data from a national survey in 33 Indonesian provinces (Basic Health Research /Riskesdas 2013) were analyzed. Multiple logistic regression models were used to calculate the odds ratio (OR) adjusted with all variables, such as age, gender, residency, education level, physical activity, and food intake. An urban-rural residence difference was found in the factors related to obesity. Daily caffeinated soft drinks and energy drinks consumption (OR = 1.12, 95% CI: 1.01-1.23) were related to risk of overweight and obesity in urban areas. Daily grilled foods (OR = 1.32, 95% CI: 1.22-1.42) and salty food (OR = 1.09, 95% CI: 1.04-1.15) consumption were significantly associated with obesity in rural areas but not in urban areas. Furthermore, sedentary activity was correlated with overweight and obesity among those who lived in urban and rural areas. Our findings suggest that education, environmental, and policy interventions may need to specifically target urban settings, where access is high to a wide range of processed and traditional high-sugar, high-fat snack foods and beverages.


Comparative evaluation of various total antioxidant capacity assays applied to phenolic compounds with the CUPRAC assay.
It would be desirable to establish and standardize methods that can measure the total antioxidant capacity level directly from vegetable extracts containing phenolics. Antioxidant capacity assays may be broadly classified as electron transfer (ET)- and hydrogen atom transfer (HAT)-based assays. The majority of HAT assays are kinetics-based, and involve a competitive reaction scheme in which antioxidant and substrate compete for peroxyl radicals thermally generated through the decomposition of azo compounds. ET-based assays measure the capacity of an antioxidant in the reduction of an oxidant, which changes colour when reduced. ET assays include the ABTS/TEAC, CUPRAC, DPPH, Folin-Ciocalteu and FRAP methods, each using different chromogenic redox reagents with different standard potentials. This review intends to offer a critical evaluation of existing antioxidant assays applied to phenolics, and reports the development by our research group of a simple and low-cost antioxidant capacity assay for dietary polyphenols, vitamins C and E, and human serum antioxidants, utilizing the copper(II)-neocuproine reagent as the chromogenic oxidizing agent, which we haved named the CUPRAC (cupric ion reducing antioxidant capacity) method. This method offers distinct advantages over other ET-based assays, namely the selection of working pH at physiological pH (as opposed to the Folin and FRAP methods, which work at alkaline and acidic pHs, respectively), applicability to both hydrophilic and lipophilic antioxidants (unlike Folin and DPPH), completion of the redox reactions for most common flavonoids (unlike FRAP), selective oxidation of antioxidant compounds without affecting sugars and citric acid commonly contained in foodstuffs and the capability to assay -SH bearing antioxidants (unlike FRAP). Other similar ET-based antioxidant assays that we have developed or modified for phenolics are the Fe(III)- and Ce(IV)-reducing capacity methods.


Getting the Message Right: Evidence-Based Insights to Improve Organizational Return-to-Work Communication Practices.
Purpose There is an absence of evidence-based guidance to support workplace stakeholders in the effective delivery of return-to-work (RTW) messages. Our study examines the specific RTW communication practices and their impact on the management of work disability. Methods Within two large and complex healthcare organizations, semi-structured interviews were conducted with workplace stakeholders (e.g., supervisors, union representatives, disability management professionals and workers' compensation representatives) and workers who had previously experienced sickness absence related to an occupational injury or illness. For workplace stakeholders interview questions asked about their roles and responsibilities in the RTW process, and specific communication strategies and messages that were used at different phases of the RTW process. For worker participants, interview questions explored RTW experiences and the impact of communication on work re-integration. An interpretative descriptive approach was used to inductively examine themes from interviews to create ways of understanding phenomena that yielded applied findings. Results Forty participants were interviewed including workplace stakeholders and workers. Participants frequently described effective RTW communication as messages that were delivered by a workplace stakeholder that included the content required by an injured worker to navigate the organizational disability management process and utilized specific strategies to address the perceived attitudes and perceptions held by an injured worker regarding work re-integration. Workplace stakeholders described five specific communication strategies including relaying messages of support, optimizing the timing of communication, careful word choice, framing messages, and tailoring communication to the injured worker. Conclusion RTW communication is an active process that requires a strategic approach. Effective communication practices represent an important strategy for workplace stakeholders to address the barriers held by injured workers and foster early and sustained RTW.


Predictive value of the pattern electroretinogram in high-risk ocular hypertension.
The pattern electroretinogram (PERG) is reduced in early glaucoma and in some eyes with ocular hypertension (OHT). This study was designed to test the ability of the PERG to predict the development of visual field loss in OHT eyes. In 29 eyes of 18 patients with high-risk OHT, the visual fields were tested, and the PERG was recorded using counterphasing checkerboard patterns at check sizes of 0.8 degrees and 15 degrees at 16 reversals/sec. The visual fields were retested 11-31 mo later. Initially, 12 eyes had pathologic PERG recordings. During the observation period, visual field loss developed in five eyes. All these eyes had a pathologic PERG recorded during the first visit. Of the eyes that had a normal PERG, none had field changes. Within the reservations of the limited sample size and follow-up interval, these findings suggest that the PERG can be used to discriminate between patients with OHT who will develop visual field loss and those who will not.


Effect of synergistic wrist motion on adhesion formation after repair of partial flexor digitorum profundus tendon lacerations in a canine model in vivo.
Therapy employing passive finger flexion and active finger extension with the wrist fixed in flexion is commonly used after flexor tendon repair. However, this method of rehabilitation may not produce full tendon excursion because of buckling of the tendon within its sheath with passive flexion. Studies of cadavera suggest that the use of synergistic wrist and finger motion may improve tendon gliding. The purpose of this study was to assess the effects of passive digital motion, performed with either wrist fixation or synergistic wrist motion, on adhesion and gap formation after flexor tendon repair. Sixty-six dogs were randomly allocated to two groups. In each group, two flexor digitorum profundus tendons of one forepaw were partially (80%) lacerated and then repaired with a modified Kessler suture. In each group, a different postoperative therapy (wrist fixation or synergistic motion) was performed twice daily. The dogs were killed at one week, three weeks, or six weeks after surgery, and the repaired tendons were evaluated to determine the adhesion grade and adhesion breaking strength. The synergistic motion group had a significantly lower adhesion grade and significantly less adhesion breaking strength than the wrist fixation group at three and six weeks (p < 0.05). At one week, there was no significant difference between the two therapy groups (p > 0.05). Passive digital flexion and extension with synergistic wrist motion was an effective therapy after repair of partial zone-2 lacerations in a canine model.


Nuclear gene targeting in Chlamydomonas using engineered zinc-finger nucleases.
The unicellular green alga Chlamydomonas reinhardtii is a versatile model for fundamental and biotechnological research. A wide range of tools for genetic manipulation have been developed for this alga, but specific modification of nuclear genes is still not routinely possible. Here, we present a nuclear gene targeting strategy for Chlamydomonas that is based on the application of zinc-finger nucleases (ZFNs). Our approach includes (i) design of gene-specific ZFNs using available online tools, (ii) evaluation of the designed ZFNs in a Chlamydomonas in situ model system, (iii) optimization of ZFN activity by modification of the nuclease domain, and (iv) application of the most suitable enzymes for mutagenesis of an endogenous gene. Initially, we designed a set of ZFNs to target the COP3 gene that encodes the light-activated ion channel channelrhodopsin-1. To evaluate the designed ZFNs, we constructed a model strain by inserting a non-functional aminoglycoside 3'-phosphotransferase VIII (aphVIII) selection marker interspaced with a short COP3 target sequence into the nuclear genome. Upon co-transformation of this recipient strain with the engineered ZFNs and an aphVIII DNA template, we were able to restore marker activity and select paromomycin-resistant (Pm-R) clones with expressing nucleases. Of these Pm-R clones, 1% also contained a modified COP3 locus. In cases where cells were co-transformed with a modified COP3 template, the COP3 locus was specifically modified by homologous recombination between COP3 and the supplied template DNA. We anticipate that this ZFN technology will be useful for studying the functions of individual genes in Chlamydomonas.


Identification of a bacterial pathogen associated with Porites white patch syndrome in the Western Indian Ocean.
Porites white patch syndrome (PWPS) is a coral disease recently described in the Western Indian Ocean. This study aimed to isolate and identify potential pathogens associated with PWPS utilizing both culture and nonculture screening techniques and inoculation trials. A total of 14 bacterial strains (those dominant in disease lesions, absent or rare in healthy tissues and considered potential pathogens in a previous study) were cultured and used to experimentally inoculate otherwise healthy individuals in an attempt to fulfil Henle-Koch's postulates. However, only one (P180R), identified as closely related (99-100% sequence identity based on 1.4 kb 16S RNA sequence) to Vibrio tubiashii, elicited signs of disease in tank experiments. Following experimental infection (which resulted in a 90% infection rate), the pathogen was also successfully re-isolated from the diseased tissues and re-inoculated in healthy corals colonies, therefore fulfilling the final stages of Henle-Koch's postulates. Finally, we report that PWPS appears to be a temperature-dependent disease, with significantly higher tissue loss (anova: d.f. = 2, F = 39.77, P < 0.01) occurring at 30 °C [1.45 ± 0.85 cm(2) per day (mean ± SE)] compared to ambient temperatures of 28 and 26 °C (0.73 ± 0.80 cm(2) per day (mean ± SE) and 0.51 ± 0.50 cm(2) per day (mean ± SE), respectively).


Divergent Isoprenoid Biosynthesis Pathways in Staphylococcus Species Constitute a Drug Target for Treating Infections in Companion Animals.
Staphylococcus species are a leading cause of skin and soft tissue infections in humans and animals, and the antibiotics used to treat these infections are often the same. Methicillin- and multidrug-resistant staphylococcal infections are becoming more common in human and veterinary medicine. From a "One Health" perspective, this overlap in antibiotic use and resistance raises concerns over the potential spread of antibiotic resistance genes. Whole-genome sequencing and comparative genomics analysis revealed that Staphylococcus species use divergent pathways to synthesize isoprenoids. Species frequently associated with skin and soft tissue infections in companion animals, including S. schleiferi and S. pseudintermedius, use the nonmevalonate pathway. In contrast, S. aureus, S. epidermidis, and S. lugdunensis use the mevalonate pathway. The antibiotic fosmidomycin, an inhibitor of the nonmevalonate pathway, was effective in killing canine clinical staphylococcal isolates but had no effect on the growth or survival of S. aureus and S. epidermidis. These data identify an essential metabolic pathway in Staphylococcus that differs among members of this genus and suggest that drugs such as fosmidomycin, which targets enzymes in the nonmevalonate pathway, may be an effective treatment for certain staphylococcal infections. IMPORTANCE Drug-resistant Staphylococcus species are a major concern in human and veterinary medicine. There is a need for new antibiotics that exhibit a selective effect in treating infections in companion and livestock animals and that would not be used to treat human bacterial infections. We have identified fosmidomycin as an antibiotic that selectively targets certain Staphylococcus species that are often encountered in skin infections in cats and dogs. These findings expand our understanding of Staphylococcus evolution and may have direct implications for treating staphylococcal infections in veterinary medicine.


Drug therapy increases bone density in osteonecrosis of the femoral head in canines.
Although many different pathogenetic mechanisms have been proposed for osteonecrosis of the femoral head, the repair process leads to structural collapse when bone resorption exceeds production. The purpose of the current study was to determine the effects of two agents with known bone-altering qualities, alendronate and simvastatin, on the healing response of a cryosurgically induced necrotic lesion of the femoral head in canines. Eighteen beagles had cryosurgical necrosis of the right femoral head. After 2 weeks, in a blinded, placebo-controlled, randomized fashion, a 10-mg dose of oral alendronate (n = 6), a 40 mg dose of simvastatin (n = 6), or a placebo (n = 6) was administered daily for 12 weeks. At sacrifice, bone densitometry and histomorphometry quantified bone in the femoral head. In the alendronate-treated animals, a 16% increase in bone mineral density of the femoral head with induced osteonecrosis was found compared with the placebo group. Increases in bone volume and trabecular thickness also were detected in the alendronate and simvastatin groups, with alendronate having the greatest effect. Clinically, increasing the amount of bone in the femoral head may forestall mechanisms leading to joint collapse characteristic of advanced osteonecrosis of the femoral head.


Purification and characterization of a novel antifungal protein with antiproliferation and anti-HIV-1 reverse transcriptase activities from Peganum harmala seeds.
A novel antifungal protein, designated as PHP, was isolated from the seeds of Peganum harmala, by cationic exchange chromatography on Resource S column and gel filtration on Sephadex 75 10/300 GL column. PHP was found to form a homodimer of about 16 kDa. Isoelectric focusing polyacrylamide gel electrophoresis analysis showed that the isoelectric point of PHP was ∼8.4. The N-terminal 20-amino acid sequence of PHP, ITCPQVTQSLAPCVPYLISG, resembles the non-specific lipid transfer proteins in certain plants. PHP exhibited lipid-binding activity. Furthermore, PHP exerted antifungal activity against Alternaria alternate, Penicillium degitatum, Rhizopus stuolonifer, and Magnaporthe grisea, and its antifungal activity was stable in the temperature range 4-60°C, and in the pH range 4-10. It inhibited the mycelial growth in A. alternate, P. degitatum, R. stuolonifer, and M. grisea with 50% inhibitory concentration (IC(50)) of 1.5, 37.5, 8.44, and 12.19 μM, respectively. PHP was also able to inhibit the proliferation of esophagus carcinoma (Eca-109), cervical carcinoma (HeLa), gastric carcinoma (MGC-7), and melanoma (B16) cells with IC(50) of 0.7, 2.74, 3.13, and 1.47 μM, respectively. Moreover, PHP significantly inhibited HIV-1 reverse transcriptase (RT) with an IC(50) of 1.26 μM. It did not have hemagglutinating and antibacterial activities. In conclusion, a novel antifungal protein with antiproliferation and anti-HIV-1 RT activities was obtained from P. harmala seeds.


Investigation of factors influencing the immunogenicity of hCG as a potential cancer vaccine.
Human chorionic gonadotrophin (hCG) and its β-subunit (hCGβ) are tumour autocrine growth factors whose presence in the serum of cancer patients has been linked to poorer prognosis. Previous studies have shown that vaccines which target these molecules and/or the 37 amino acid C-terminal hCGβ peptide (hCGβCTP) induce antibody responses in a majority of human recipients. Here we explored whether the immunogenicity of vaccines containing an hCGβ mutant (hCGβR68E, designed to eliminate cross-reactivity with luteinizing hormone) or hCGβCTP could be enhanced by coupling the immunogen to different carriers [keyhole limpet haemocyanin (KLH) or heat shock protein 70 (Hsp70)] using different cross-linkers [1-ethyl-3(3-dimethylaminopropyl)carboiimide (EDC) or glutaraldehyde (GAD)] and formulated with different adjuvants (RIBI or Montanide ISA720). While there was little to choose between KLH and Hsp70 as carriers, their influence on the effectiveness of a vaccine containing the BAChCGβR68E mutant was less marked, presumably because, being a foreign species, this mutant protein itself might provide T helper epitopes. The mutant provided a significantly better vaccine than the hCGβCTP peptide irrespective of the carrier used, how it was cross-linked to the carrier or which adjuvant was used when hCG was the target. Nonetheless, for use in humans where hCG is a tolerated self-protein, the need for a carrier is of fundamental importance. Highest antibody titres were obtained by linking the BAChCGβR68E to Hsp70 as a carrier by GAD and using RIBI as the adjuvant, which also resulted in antibodies with significantly higher affinity than those elicited by hCGβCTP peptide vaccine. This makes this mutant vaccine a promising candidate for therapeutic studies in hCGβ-positive cancer patients.


Costly but worthless gifts facilitate courtship.
What are the characteristics of a good courtship gift? We address this question by modelling courtship as a sequential game. This is structured as follows: the male offers a gift to a female; after observing the gift, the female decides whether or not to accept it; she then chooses whether or not to mate with the male. In one version of the game, based on human courtship, the female is uncertain about whether the male intends to stay or desert after mating. In a second version, there is no paternal care but the female is uncertain about the male's quality. The two versions of the game are shown to be mathematically equivalent. We find robust equilibrium solutions in which mating is predominantly facilitated by an "extravagant" gift which is costly to the male but intrinsically worthless to the female. By being costly to the male, the gift acts as a credible signal of his intentions or quality. At the same time, its lack of intrinsic value to the female serves to deter a "gold-digger", who has no intention of mating with the male, from accepting the gift. In this way, an economically inefficient gift enables mutually suitable partners to be matched.


Language-association cortex asymmetry in autism and specific language impairment.
Language deficits are among the core impairments of autism. We previously reported asymmetry reversal of frontal language cortex in boys with autism. Specific language impairment (SLI) and autism share similar language deficits and may share genetic links. This study evaluated asymmetry of frontal language cortex in a new, independent sample of right-handed boys, including a new sample of boys with autism and a group of boys with SLI. The boys with autism were divided into those with language impairment (ALI) and those with normal language ability (ALN). Subjects (right-handed, aged 6.2-13.4 years) included 22 boys with autism (16 ALI and 6 ALN), 9 boys with a history of or present SLI, and 11 normal controls. MRI brain scans were segmented into grey and white matter; then the cerebral cortex was parcellated into 48 gyral-based divisions per hemisphere. Group differences in volumetric asymmetry were predicted a priori in language-related regions in inferior lateral frontal (Broca's area) and posterior superior temporal cortex. Language impaired boys with autism and SLI both had significant reversal of asymmetry in frontal language-related cortex; larger on the right side in both groups of language impaired boys and larger on the left in both unimpaired language groups, strengthening a phenotypic link between ALI and SLI. Thus, we replicated the observation of reversed asymmetry in frontal language cortex reported previously in an independent autism sample, and observed similar reversal in boys with SLI, further strengthening a phenotypic link between SLI and a subgroup of autism. Linguistically unimpaired boys with autism had similar asymmetry compared with the control group, suggesting that Broca's area asymmetry reversal is related more to language impairment than specifically to autism diagnosis.


New World Cactaceae Plants Harbor Diverse Geminiviruses.
The family Cactaceae comprises a diverse group of typically succulent plants that are native to the American continent but have been introduced to nearly all other continents, predominantly for ornamental purposes. Despite their economic, cultural, and ecological importance, very little research has been conducted on the viral community that infects them. We previously identified a highly divergent geminivirus that is the first known to infect cacti. Recent research efforts in non-cultivated and asymptomatic plants have shown that the diversity of this viral family has been under-sampled. As a consequence, little is known about the effects and interactions of geminiviruses in many plants, such as cacti. With the objective to expand knowledge on the diversity of geminiviruses infecting cacti, we used previously acquired high-throughput sequencing results to search for viral sequences using BLASTx against a viral RefSeq protein database. We identified two additional sequences with similarity to geminiviruses, for which we designed abutting primers and recovered full-length genomes. From 42 cacti and five scale insects, we derived 42 complete genome sequences of a novel geminivirus species that we have tentatively named Opuntia virus 2 (OpV2) and 32 genomes of an Opuntia-infecting becurtovirus (which is a new strain of the spinach curly top Arizona virus species). Interspecies recombination analysis of the OpV2 group revealed several recombinant regions, in some cases spanning half of the genome. Phylogenetic analysis demonstrated that OpV2 is a novel geminivirus more closely related to viruses of the genus Curtovirus, which was further supported by the detection of three recombination events between curtoviruses and OpV2. Both OpV2 and Opuntia becurtoviruses were identified in mixed infections, which also included the previously characterized Opuntia virus 1. Viral quantification of the co-infected cactus plants compared with single infections did not show any clear trend in viral dynamics that might be associated with the mixed infections. Using experimental Rhizobium-mediated inoculations, we found that the initial accumulation of OpV2 is facilitated by co-infection with OpV1. This study shows that the diversity of geminiviruses that infect cacti is under-sampled and that cacti harbor diverse geminiviruses. The detection of the Opuntia becurtoviruses suggests spill-over events between viruses of cultivated species and native vegetation. The threat this poses to cacti needs to be further investigated.


Trajectories of Suicidal Ideation from Adolescence to Adulthood: Does the History of Same-Sex Experience Matter?
The fluidity of same-sex experience (SSE) can be conceptualized as both a risk and a resource that impacts suicidal ideation over time. Considering whether SSE occurs in adolescence, adulthood, or both, this study suggests that SSE in both adolescence and adulthood is associated with depression and low self-esteem throughout the life course, resulting in chronic susceptibility to suicidal ideation. Yet due to variation in both accumulation of risk and resources over time, trajectories of suicidal ideation during the transition to adulthood likely vary by timing of SSE. To test these hypotheses, we fit latent growth curve models to a gender-stratified sample taken from the National Longitudinal Study of Adolescent to Adult Health (7608 men, 8070 women). We created four groups of SSE: no SSE (6322 men, 5981 women), SSE in adolescence only (634 men, 480 women), SSE in adulthood only (372 men, 1081 women), and SSE in adolescence and adulthood (280 men, 528 women). Men and women with SSE in both life stages had the greatest risk of suicidal ideation in adolescence and in adulthood. Yet women with first SSE in adulthood had less of a decline in suicidal ideation over time, relative to those with no SSE and those with SSE in adolescence only, and this was partially due to higher depression and lower self-esteem. Results suggest greater support is needed for adolescents expressing non-normative sexualities and for those with first SSE in adulthood, a group that is more difficult to identify in schools.


The role of evidence in the decline of antibiotic use for common respiratory infections in primary care.
Antibiotic prescribing in primary care for common respiratory infections increased steadily until the mid 1990s, when the trend reversed noticeably. During the subsequent decade, antibiotic prescribing reduced by up to one-third in some countries. Explanations for this reduction have focused on a decline in the incidence and severity of common respiratory infections, and on the resulting decrease in the number of patients seeking consultation. We argue that evidence from primary-care research had a central role in changing the practice of antibiotic prescribing, and discuss the concern that has arisen among some physicians around this issue. Targeted reductions in antibiotic prescribing constitute a balancing act between individual and societal concerns, pitting the expected gains in preserving the usefulness of an antibiotic against any given reduction in use. There may be unintended consequences for decreasing antibiotic use beyond a certain point without adequate supporting evidence. A new approach to antibiotic prescribing requires comprehensive research to answer why change is necessary, and how that change can be safely implemented. Future policies must move beyond a "one size fits all" mindset if public and provider behaviours are expected to become more congruent with the growing research evidence.


Randomized soil survey of the distribution of Burkholderia pseudomallei in rice fields in Laos.
Melioidosis is a major cause of morbidity and mortality in Southeast Asia, where the causative organism (Burkholderia pseudomallei) is present in the soil. In the Lao People's Democratic Republic (Laos), B. pseudomallei is a significant cause of sepsis around the capital, Vientiane, and has been isolated in soil near the city, adjacent to the Mekong River. We explored whether B. pseudomallei occurs in Lao soil distant from the Mekong River, drawing three axes across northwest, northeast, and southern Laos to create nine sampling areas in six provinces. Within each sampling area, a random rice field site containing a grid of 100 sampling points each 5 m apart was selected. Soil was obtained from a depth of 30 cm and cultured for B. pseudomallei. Four of nine sites (44%) were positive for B. pseudomallei, including all three sites in Saravane Province, southern Laos. The highest isolation frequency was in east Saravane, where 94% of soil samples were B. pseudomallei positive with a geometric mean concentration of 464 CFU/g soil (95% confidence interval, 372 to 579 CFU/g soil; range, 25 to 10,850 CFU/g soil). At one site in northwest Laos (Luangnamtha), only one sample (1%) was positive for B. pseudomallei, at a concentration of 80 CFU/g soil. Therefore, B. pseudomallei occurs in Lao soils beyond the immediate vicinity of the Mekong River, alerting physicians to the likelihood of melioidosis in these areas. Further studies are needed to investigate potential climatic, soil, and biological determinants of this heterogeneity.


Activation loop targeting strategy for design of receptor-interacting protein kinase 2 (RIPK2) inhibitors.
Development of selective kinase inhibitors remains a challenge due to considerable amino acid sequence similarity among family members particularly in the ATP binding site. Targeting the activation loop might offer improved inhibitor selectivity since this region of kinases is less conserved. However, the strategy presents difficulties due to activation loop flexibility. Herein, we report the design of receptor-interacting protein kinase 2 (RIPK2) inhibitors based on pan-kinase inhibitor regorafenib that aim to engage basic activation loop residues Lys169 or Arg171. We report development of CSR35 that displayed >10-fold selective inhibition of RIPK2 versus VEGFR2, the target of regorafenib. A co-crystal structure of CSR35 with RIPK2 revealed a resolved activation loop with an ionic interaction between the carboxylic acid installed in the inhibitor and the side-chain of Lys169. Our data provides principle feasibility of developing activation loop targeting type II inhibitors as a complementary strategy for achieving improved selectivity.


Hyperbaric oxygen therapy in orthopedic conditions: an evaluation of safety.
Because of the documented cellular and biochemical benefits of hyperbaric oxygen (HBO), HBO therapy is applied now with increasing frequency to various orthopedic conditions. Many traumatologists and orthopedic surgeons might refer their patients for adjuvant HBO therapy. However, the potential risks and risk-benefit ratio have often been underemphasized in therapeutic trials. From October 2002 to September 2004, 240 patients with a total of 4,638 treatments received HBO therapy at the hyperbaric medicine center of our institution on an identical treatment protocol. HBO therapy patient treatment logs were reviewed to analyze the incidence of complications during HBO treatment. The overall incidence of complications was 1.83%. Over 94% of treatment complications were mild to moderate and designated as minor complications; fewer than 6% were severe or life threatening and designated as major complications. The incidence of major complications (central nervous system [CNS] oxygen toxicity in this series) was 0.109%. There was no mortality. Two patients with unusual presentation of CNS oxygen toxicity were observed during the study period. HBO therapy in orthopedic conditions is considered as a safe treatment because of a very low complication rate; however, analysis of patients with CNS oxygen toxicity revealed its unpredictability and inevitability. Although it is common sense that patients who develop a seizure in the hospital need help from the medical staff, it cannot be done in a monoplace hyperbaric chamber because of pressure unequalization. Therefore, a multiplace chamber equipped with an antechamber for medical contingency is possibly the better facility in consideration of safety.


Coffee consumption and risk of hypertension in the Polish arm of the HAPIEE cohort study.
Coffee consumption has been hypothesized to be associated with blood pressure (BP), but previous findings are not homogeneous. The aim of this study was to evaluate the association between coffee consumption and the risk of developing hypertension. Data on coffee consumption, BP and use of anti-hypertensive medicament were derived from 2725 participants of the Polish arm of the HAPIEE project (Health, Alcohol and Psychosocial factors In Eastern Europe) who were free of hypertension at baseline and followed up for an average of 5 years. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated by multivariate logistic regression analyses and stratified for potential confounding factors. Coffee consumption was related to decreased age, smoking status and total energy intake. Compared with persons who drink <1 cup coffee per day, systolic BP was significantly associated with coffee consumption and the risk of hypertension was lower for individuals consuming 3-4 cups per day. Despite the analysis stratified by gender showed that the protective effect of coffee consumption on hypertension was significant only in women, the analysis after stratification by smoking status revealed a decreased risk of hypertension in non-smokers drinking 3-4 cups of coffee per day in both sexes (OR 0.41, 95% CI: 0.21, 0.79 for men and OR 0.54, 95% CI: 0.29, 0.99 for women). Upper category coffee consumption (>4 cups per day) was not related to significant increased risk of hypertension. Relation between coffee consumption and incidence of hypertension was related to smoking status. Consumption of 3-4 cups of coffee per day decreased the risk of hypertension in non-smoking men and women only.


Selectively Inhibiting the Median Preoptic Nucleus Attenuates Angiotensin II and Hyperosmotic-Induced Drinking Behavior and Vasopressin Release in Adult Male Rats.
The median preoptic nucleus (MnPO) is a putative integrative region that contributes to body fluid balance. Activation of the MnPO can influence thirst, but it is not clear how these responses are linked to body fluid homeostasis. We used designer receptors exclusively activated by designer drugs (DREADDs) to determine the role of the MnPO in drinking behavior and vasopressin release in response to peripheral angiotensin II (ANG II) or 3% hypertonic saline (3% HTN) in adult male Sprague Dawley rats (250-300 g). Rats were anesthetized with isoflurane and stereotaxically injected with an inhibitory DREADD (rAAV5-CaMKIIa-hM4D(Gi)-mCherry) or control (rAAV5-CaMKIIa-mCherry) virus in the MnPO. After two weeks' recovery, a subset of rats was used for extracellular recordings to verify functional effects of ANG II or hyperosmotic challenges in MnPO slice preparations. Remaining rats were used in drinking behavior studies. Each rat was administered either 10 mg/kg of exogenous clozapine-N-oxide (CNO) to inhibit DREADD-expressing cells or vehicle intraperitoneal followed by a test treatment with either 2-mg/kg ANG II or 3% HTN (1 ml/100-g bw, s.c.), twice per week for two separate treatment weeks. CNO-induced inhibition during either test treatment significantly attenuated drinking responses compared to vehicle treatments and controls. Brain tissue processed for cFos immunohistochemistry showed decreased expression with CNO-induced inhibition during either test treatment in the MnPO and downstream nuclei compared to controls. CNO-mediated inhibition significantly attenuated treatment-induced increases in plasma vasopressin compared to controls. The results indicate inhibition of CaMKIIa-expressing MnPO neurons significantly reduces drinking and vasopressin release in response to ANG II or hyperosmotic challenge.


Is pricing of dolutegravir equitable? A comparative analysis of price and country income level in 52 countries.
Differences in marketed prices of antiretrovirals raises questions about the fairness of pricing medicines of significant public health importance such as dolutegravir (DTG). In view of the reduced prices of generically available efavirenz (EFV), there is a need to determine if previous conclusions on DTG's cost-effectiveness need to be re-assessed. Lowest list prices of DTG were extracted from national drug price or reimbursement databases for 52 countries. Price was recorded as US$ per person-year (ppy). We compared the price of DTG to minimum costs of production and reduced prices of EFV, as well as assessed the correlation with gross domestic product (GDP) per capita and HIV epidemic size in three income classification groups ('high', 'upper middle-income', 'lower middle or low-income'). Annual prices of DTG ranged from $27 per person-year in Georgia to $20,130 in the USA. Within each income group, there was no observable relationship between DTG prices, GDP per capita and HIV epidemic size. Median price in countries excluded from voluntary licensing agreements ($8718) was >140 times higher than countries included ($60). Price of DTG was >500% higher than EFV in many countries. Three full economic evaluations from high-income settings that compared DTG against EFV all used branded drug prices of EFV-based regimens as cost inputs to evaluate DTG's cost-effectiveness. This study highlights the wide disparity in prices of DTG across countries, even when segregated by similar income levels. The cost-effectiveness of DTG versus EFV should be re-evaluated now that low-cost generic EFV has become widely available.


An anthropomorphic abdominal phantom for deformable image registration accuracy validation in adaptive radiation therapy.
To design and construct a three-dimensional (3D) anthropomorphic abdominal phantom for geometric accuracy and dose summation accuracy evaluations of deformable image registration (DIR) algorithms for adaptive radiation therapy (ART). Organ molds, including liver, kidney, spleen, stomach, vertebra, and two metastasis tumors, were 3D printed using contours from an ovarian cancer patient. The organ molds were molded with deformable gels made of different mixtures of polyvinyl chloride (PVC) and the softener dioctyl terephthalate. Gels with different densities were obtained by a polynomial fitting curve that described the relation between the Hounsfield unit (HU) and PVC-softener blending ratio. The rigid vertebras were constructed by molding of white cement and cellulose pulp. The final abdominal phantom was assembled by arranging all the fabricated organs inside a hollow dummy according to their anatomies, and sealed by deformable gel with averaged HU of muscle and fat. Fiducial landmarks were embedded inside the phantom for spatial accuracy and dose accumulation accuracy studies. Two channels were excavated to facilitate ionization chamber insertion for dosimetric measurements. Phantom properties such as deformable gel elasticity and HU stability were studied. The dosimetric measurement accuracy in the phantom was performed, and the DIR accuracies of three DIR algorithms available in the open source DIR toolkit-DIRART were also validated. The constructed deformable gel showed elastic behavior and was stable in HU values over times, proving to be a practical material for the deformable phantom. The constructed abdominal phantom consisted of realistic anatomies in terms of both anatomical shapes and densities when compared with its reference patient. The dosimetric measurements showed a good agreement with the calculated doses from the treatment planning system. Fiducial-based accuracy analysis conducted on the constructed phantom demonstrated the feasibility of applying the phantom for organ-wise DIR accuracy assessment. We have designed and constructed an anthropomorphic abdominal deformable phantom with satisfactory elastic property, realistic organ density, and anatomy. This physical phantom can be used for routine validations of DIR geometric accuracy and dose accumulation accuracy in ART.


Regulated CPEB phosphorylation during meiotic progression suggests a mechanism for temporal control of maternal mRNA translation.
CPEB is an mRNA-binding protein that stimulates polyadenylation-induced translation of maternal mRNA once it is phosphorylated on Ser 174 or Thr 171 (species-dependent). Disruption of the CPEB gene in mice causes an arrest of oogenesis at embryonic day 16.5 (E16.5), when most oocytes are in pachytene of prophase I. Here, we show that CPEB undergoes Thr 171 phosphorylation at E16.5, but dephosphorylation at the E18.5, when most oocytes are entering diplotene. Although phosphorylation is mediated by the kinase aurora, the dephosphorylation is due to the phosphatase PP1. The temporal control of CPEB phosphorylation suggests a mechanism in which CPE-containing mRNA translation is stimulated at pachytene and metaphase I.


Estrogen sulfotransferase and sulfatase: Roles in the regulation of estrogen activity in human uterine endometrial carcinomas.
The regulation of estrogen activity through the formation and cleavage of sulfoconjugates of estrogens is known to be related to the progression and metastasis of estrogen-dependent breast carcinomas, but the involvement of sulfoconjugates in the steroid stimulation of endometrial functions and the progression of endometrial adenocarcinomas is not clearly understood yet. Estrogen sulfotransferase (EST) in the uterine endometria during the follicular phase was more active than during the luteal phase, but estrogen sulfate (ES) sulfatase exhibited lower activity during the follicular phase than during the luteal phase. However, ES sulfatase activities in cancerous tissues were lower than those in normal endometria and endometrial adenocarcinoma-derived cells, among which the activity was exceedingly high in Ishikawa cells, suggesting that ES sulfatase in Ishikawa cells contributes to the estrogen-dependent growth of these cells. EST activities higher than that in Ishikawa cells were found in only 3 of 24 cancerous tissues. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of the EST and ES sulfatase genes in carcinoma-derived cells demonstrated the extensive expression of both genes in Ishikawa cells. The isolated EST gene was transfected into Ishikawa cells with a mammalian expression vector to establish cell clones with enhanced EST activity, and the estrogen-dependent cell growth of the resultant cell clones was found to be abolished, due to the enhanced sulfoconjugation of estrogen. Since ES sulfatase activity in cancerous tissues was significantly lower than that in Ishikawa cells, it might be not involved in the enhancement of estrogen activity associated with the pathogenesis of endometrial adenocarcinoma tissues.


Single-cell phenotyping within transparent intact tissue through whole-body clearing.
Understanding the structure-function relationships at cellular, circuit, and organ-wide scale requires 3D anatomical and phenotypical maps, currently unavailable for many organs across species. At the root of this knowledge gap is the absence of a method that enables whole-organ imaging. Herein, we present techniques for tissue clearing in which whole organs and bodies are rendered macromolecule-permeable and optically transparent, thereby exposing their cellular structure with intact connectivity. We describe PACT (passive clarity technique), a protocol for passive tissue clearing and immunostaining of intact organs; RIMS (refractive index matching solution), a mounting media for imaging thick tissue; and PARS (perfusion-assisted agent release in situ), a method for whole-body clearing and immunolabeling. We show that in rodents PACT, RIMS, and PARS are compatible with endogenous-fluorescence, immunohistochemistry, RNA single-molecule FISH, long-term storage, and microscopy with cellular and subcellular resolution. These methods are applicable for high-resolution, high-content mapping and phenotyping of normal and pathological elements within intact organs and bodies.


BINOL-3,3'-triflone N,N-dimethyl phosphoramidites: through-space 19F,31P spin-spin coupling with a remarkable dependency on temperature and solvent internal pressure.
A combined computational and experimental study of the effects of solvent, temperature and stereochemistry on the magnitude of the through-space spin-spin coupling between 31P and 19F nuclei which are six-bonds apart is described. The reaction of 3-trifluoromethylsulfonyl-2,'2-dihydroxy-1,1'-binaphthalene (3-SO2CF3-BINOL) with hexamethylphosphorous triamide (P(NMe2)3) generates a pair of N,N-dimethylphosphoramidites which are diastereomeric due to their differing relative configurations at the stereogenic phosphorous centre and the axially chiral (atropisomeric) BINOL unit. Through-space NMR coupling of the 31P and 19F nuclei of the phosphoramidite and sulfone is detected in one diastereomer only. In the analogous N,N-dimethylphosphoramidite generated from 3,3'-(SO2CF3)2-BINOL only one of the diastereotopic trifluoromethylsulfone moieties couples with the 31P of the phosphoramidite. In both cases, the magnitude of the coupling is strongly modulated (up to 400 %) by solvent and temperature. A detailed DFT analysis of the response of the coupling to the orientation of the CF3 moiety with respect to the P-lone pair facilitates a confident assignment of the stereochemical identity of the pair of diastereomers. The analysis shows that the intriguing effects of environment on the magnitude of the coupling can be rationalised by a complex interplay of solvent internal pressure, molecular volume and thermal access to a wider conformational space. These phenomena suggest the possibility for the design of sensitive molecular probes for local environment that can be addressed via through-space NMR coupling.


The occurrence of priapism as a result of the use of a single dose of quetiapine.
Priapism is a prolonged pathologic erection situation that occurs after sexual stimulation or without sexual stimulation. This condition is divided into two types, ischemic (low-flow, veno-occlusive) and non-ischemic (high-flow, arterial). A 68-year-old male patient applied to our clinic with the complaints of hardness and pain in the penis. Three days before he applied to our clinic, he was prescribed a single dose of 200 mg quetiapine by a psychiatry polyclinic for the complication of insomnia. Nearly 6 hours after the first dose of quetiapine, an involuntary erection occurred with accompanying pain in the penis. The patient waited for spontaneous detumescence without consulting a psychiatrist. After the patient had waited for 48 hours without any change, he applied to our clinic. Other etiologic factors of priapism were excluded (such as malignancy, blood dyscrasia, leukemia, and trauma). In blood gas samples obtained from the corpus cavernosum, hypoxia, hypercarbia, and acidosis were diagnosed. Ischemic priapism was supposed. We conclude that priapism can be viewed as a new possible side effect of quetiapine and that patients should be warned about this side effect.


Periodic Variation of Mutation Rates in Bacterial Genomes Associated with Replication Timing.
The causes and consequences of spatiotemporal variation in mutation rates remain to be explored in nearly all organisms. Here we examine relationships between local mutation rates and replication timing in three bacterial species whose genomes have multiple chromosomes: Vibrio fischeri, Vibrio cholerae, and Burkholderia cenocepacia Following five mutation accumulation experiments with these bacteria conducted in the near absence of natural selection, the genomes of clones from each lineage were sequenced and analyzed to identify variation in mutation rates and spectra. In lineages lacking mismatch repair, base substitution mutation rates vary in a mirrored wave-like pattern on opposing replichores of the large chromosomes of V. fischeri and V. cholerae, where concurrently replicated regions experience similar base substitution mutation rates. The base substitution mutation rates on the small chromosome are less variable in both species but occur at similar rates to those in the concurrently replicated regions of the large chromosome. Neither nucleotide composition nor frequency of nucleotide motifs differed among regions experiencing high and low base substitution rates, which along with the inferred ~800-kb wave period suggests that the source of the periodicity is not sequence specific but rather a systematic process related to the cell cycle. These results support the notion that base substitution mutation rates are likely to vary systematically across many bacterial genomes, which exposes certain genes to elevated deleterious mutational load.IMPORTANCE That mutation rates vary within bacterial genomes is well known, but the detailed study of these biases has been made possible only recently with contemporary sequencing methods. We applied these methods to understand how bacterial genomes with multiple chromosomes, like those of Vibrio and Burkholderia, might experience heterogeneous mutation rates because of their unusual replication and the greater genetic diversity found on smaller chromosomes. This study captured thousands of mutations and revealed wave-like rate variation that is synchronized with replication timing and not explained by sequence context. The scale of this rate variation over hundreds of kilobases of DNA strongly suggests that a temporally regulated cellular process may generate wave-like variation in mutation risk. These findings add to our understanding of how mutation risk is distributed across bacterial and likely also eukaryotic genomes, owing to their highly conserved replication and repair machinery.


Arbitrarily high focal depth with finite apertures.
We modify the concept of superresolution for achieving arbitrarily extended focal depth with a finite aperture. The Legendre polynomials are used for designing a novel apodizer that produces arbitrarily high depth of focus. This kind of apodizer can also arbitrarily reduce the sensitivity to spherical aberration. Since these benefits are achieved at the expense of light throughput, we report on a formula for evaluating light throughput of this kind of filter.


Paradoxical myeloid-derived suppressor cell reduction in the bone marrow of SIV chronically infected macaques.
Myeloid derived suppressor cells (MDSCs), which suppress anti-tumor or anti-viral immune responses, are expanded in the peripheral blood and tissues of patients/animals with cancer or viral infectious diseases. We here show that in chronic SIV infection of Indian rhesus macaques, the frequency of MDSCs in the bone marrow (BM) was paradoxically and unexpectedly decreased, but increased in peripheral blood. Reduction of BM MDSCs was found in both CD14+MDSC and Lin-CD15+MDSC subsets. The reduction of MDSCs correlated with high plasma viral loads and low CD4+ T cell counts, suggesting that depletion of BM MDSCs was associated with SIV/AIDS disease progression. Of note, in SHIVSF162P4-infected macaques, which naturally control viral replication within a few months of infection, the frequency of MDSCs in the bone marrow was unchanged. To investigate the mechanisms by which BM MDSCs were reduced during chronic SIV infection, we tested several hypotheses: depletion due to viral infection, alterations in MDSC trafficking, and/or poor MDSC replenishment. We found that the possible mobilization of MDSCs from BM to peripheral tissues and the slow self-replenishment of MDSCs in the BM, along with the viral infection-induced depletion, all contribute to the observed BM MDSC reduction. We first demonstrate MDSC SIV infection in vivo. Correlation between BM CD14+MDSC reduction and CD8+ T cell activation in tissues is consistent with decreased immune suppression by MDSCs. Thus, depletion of BM MDSCs may contribute to the pathologic immune activation during chronic SIV infection and by extension HIV infection.


Psychotherapy as a treatment modality for psychiatric disorders: Perceptions of general public of Karachi, Pakistan.
Psychiatric disorders affect about 450 million individuals worldwide. A number of treatment modalities such as psychotropic medications, psychotherapy and electroconvulsive therapy can be used to treat these disorders. Attitudes of general public play a pivotal role in effective utilization of mental health services. We explored the perceptions of general public of Karachi, Pakistan regarding psychotherapy. A cross-sectional study was conducted in Karachi, Pakistan during July-August, 2008. A three-step sampling strategy and a structured questionnaire were employed to survey knowledge and perceptions of adult general public about psychotherapy. Descriptive statistics were used for baseline characteristics. Logistic regression models were used to investigate any significant associations between baseline characteristics of the participants and their perceptions. The study sample comprised of 985 individuals (536 males; 531 financially independent) with an average age of 36.7 years (SD 13.54 years) and 12.5 years (SD 3.09 years) of education were included. Majority (59.4%; n = 585) claimed to be aware of psychotherapy as a treatment option for psychiatric disorders but 47.5% of these (n = 278/585) failed to identify its correct definition. Concerns voiced by the participants about psychotherapy included stigma (48.7%) and breech in confidentiality (39.5%); 60.7% opined it cost effective and 86.5% favored its use as an adjuvant modality. A preference for psychotherapy as the treatment strategy for psychiatric disorders was demonstrated by 46.6% (n = 459/985). Younger, more educated, financially independent and female participants were more likely to prefer psychotherapy as were those who deemed it cost effective. Positive attitudes regarding the acceptability, clinical utility and cost-effectiveness of psychotherapy were observed in a sample representative of general public of Karachi, Pakistan. These findings highlight its potential utility for devising pragmatic mental health strategies in the face of limited resources.


Unsupervised Domain Adaptation on Person Reidentification via Dual-Level Asymmetric Mutual Learning.
Unsupervised domain adaptation (UDA) person reidentification (Re-ID) aims to identify pedestrian images within an unlabeled target domain with an auxiliary labeled source-domain dataset. Many existing works attempt to recover reliable identity information by considering multiple homogeneous networks. And take these generated labels to train the model in the target domain. However, these homogeneous networks identify people in approximate subspaces and equally exchange their knowledge with others or their mean net to improve their ability, inevitably limiting the scope of available knowledge and putting them into the same mistake. This article proposes a dual-level asymmetric mutual learning (DAML) method to learn discriminative representations from a broader knowledge scope with diverse embedding spaces. Specifically, two heterogeneous networks mutually learn knowledge from asymmetric subspaces through the pseudo label generation in a hard distillation manner. The knowledge transfer between two networks is based on an asymmetric mutual learning (AML) manner. The teacher network learns to identify both the target and source domain while adapting to the target domain distribution based on the knowledge of the student. Meanwhile, the student network is trained on the target dataset and employs the ground-truth label through the knowledge of the teacher. Extensive experiments in Market-1501, CUHK-SYSU, and MSMT17 public datasets verified the superiority of DAML over state-of-the-arts (SOTA).


Sexual dimorphism in insulin sensitivity and susceptibility to develop diabetes in rats.
The goal of this study was to evaluate gender-related differences of some metabolic determinants of insulin sensitivity and of susceptibility to the effects of diabetes. Changes in body weight, blood glucose, and serum insulin concentrations were compared between female and male Wistar rats in prepubertal, pubertal, and adult stages of life. A diabetic model was induced by streptozotocin (STZ) under nicotinamide protection in both sexes and metabolic patterns were evaluated during the next 4 weeks. Finally, the pancreases were processed for morphometric analysis. In the three age groups, at similar blood glucose levels, higher fasting serum insulin levels were found in female as compared with age matched male rats. After STZ treatment, female rats show lower insulin and higher glucose levels, and a worse survival rate as compared with male rats. The more severe disease phenotype observed in female animals is associated with a more dramatic perturbation of pancreatic islet morphology. Significant differences exist in insulin sensitivity between sexes, females being less sensitive to insulin than males at all age groups and more susceptible to the rapid development of a more severe form of diabetes than males.


Association between childhood maltreatment and normal adult personality traits: exploration of an understudied field.
We assessed normal personality traits and childhood trauma in approximately 1170 subjects from a general population-based community sample. In bivariate analyses emotional abuse was most pervasively related to personality, showing significant detrimental associations with neuroticism, extraversion, openness, conscientiousness, and agreeableness. Neuroticism was significantly related to emotional abuse and neglect, physical abuse and neglect, and sexual abuse. Emotional abuse was related to neuroticism in men more profoundly than in women (β = 0.095). Adjusting for the covariance between childhood maltreatment variables, neuroticism was mainly related to emotional abuse (β = 0.193), extraversion to emotional neglect (β = -0.259), openness to emotional abuse (β = 0.175), conscientiousness to emotional abuse (β = -0.110), and agreeableness to emotional neglect (β = -0.153). The proportion of variance explained was highest in neuroticism (5.6%) and lowest in openness (1.9%) and conscientiousness (1.8%). These findings help to understand the complex association between childhood maltreatment and both normal and pathological personality.


Supramolecular high-aspect ratio assemblies with strong antifungal activity.
Efficient and pathogen-specific antifungal agents are required to mitigate drug resistance problems. Here we present cationic small molecules that exhibit excellent microbial selectivity with minimal host toxicity. Unlike typical cationic polymers possessing molecular weight distributions, these compounds have an absolute molecular weight aiding in isolation and characterization. However, their specific molecular recognition motif (terephthalamide-bisurea) facilitates spontaneous supramolecular self-assembly manifesting in several polymer-like properties. Computational modelling of the terephthalamide-bisurea structures predicts zig-zag or bent arrangements where distal benzyl urea groups stabilize the high-aspect ratio aqueous supramolecular assemblies. These nanostructures are confirmed by transmission electron microscopy and atomic force microscopy. Antifungal activity against drug-sensitive and drug-resistant strains with in vitro and in vivo biocompatibility is observed. Additionally, despite repeated sub-lethal exposures, drug resistance is not induced. Comparison with clinically used amphotericin B shows similar antifungal behaviour without any significant toxicity in a C. albicans biofilm-induced mouse keratitis model.


High CO2 concentration as an inductor agent to drive production of recombinant phytotoxic antimicrobial peptides in plant biofactories.
Cationic α-helical antimicrobial peptides such as BP100 are of increasing interest for developing novel phytosanitary or therapeutic agents and products with industrial applications. Biotechnological production of these peptides in plants can be severely impaired due to the toxicity exerted on the host by high-level expression. This can be overcome by using inducible promoters with extremely low activity throughout plant development, although the yields are limited. We examined the use of modified atmospheres using the increased levels of [CO2], commonly used in the food industry, as the inductor agent to biotechnologically produce phytotoxic compounds with higher yields. Here we show that 30% [CO2] triggered a profound transcriptional response in rice leaves, including a change in the energy provision from photosynthesis to glycolysis, and the activation of stress defense mechanisms. Five genes with central roles in up-regulated pathways were initially selected and their promoters successfully used to drive the expression of phytotoxic BP100 in genetically modified (GM) rice. GM plants had a normal phenotype on development and seed production in non-induction conditions. Treatment with 30 % [CO2] led to recombinant peptide accumulation of up to 1 % total soluble protein when the Os.hb2 promoter was used. This is within the range of biotechnological production of other peptides in plants. Using BP100 as a proof-of-concept we demonstrate that very high [CO2] can be considered an economically viable strategy to drive production of recombinant phytotoxic antimicrobial peptides in plant biofactories.


Quinidine enhances and suppresses Kv1.2 from outside and inside the cell, respectively.
We studied the effects of quinidine on two K channel clones expressed in Xenopus oocytes, Kv1.2 and Kv1.4. Quinidine had an agonist (enhancement) and a blocker (suppression) action on Kv1.2, whereas only the blocker action was seen with Kv1.4. For both, the blocker action was exerted from the intracellular side of the membrane due to quinidine binding in its protonated form to a site in the membrane electrical field. The agonist action on Kv1.2 was exerted from the extracellular side of the membrane. A quaternary analog of quinidine, Q+1C, that had a low membrane permeability allowed the agonist action to be separated from the blocker action. Experiments with Q+1C showed that the agonist action was caused by a parallel shift of the voltage dependence of Kv1.2 activation in the negative direction. There was no increase in channel conductance when channel activation reached a plateau. Q+1C had little or no effect on the rate of Kv1.2 activation, but significantly slowed Kv1.2 deactivation at voltages as negative as - 160 mV. These changes in channel gating could be largely reproduced by a simple kinetic model, assuming that the agonist action was caused by a decrease in the rate constants for transitions from the activated to the rested states. In oocytes coexpressing Kv1.2 and Kv1.4 at approximately a 1:1 ratio, the response to Q+1C was similar to that of oocytes expressing Kv1.2 alone, suggesting that heteromultimer channels of Kv1.2 and Kv1.4 subunits could respond to the agonist action of quinidine.


Gadolinium(III) complexes of 1,4,7-triazacyclononane based picolinate ligands: simultaneous optimization of water exchange kinetics and electronic relaxation.
The two new tripodal picolinate H(3)ebpatcn (1-carboxyethyl-4,7-bis((6-carboxypyridin-2-yl)methyl)-1,4,7-triazacyclononane) and H(4)pbpatcn (1-methylphosphonic-acid-4,7-bis((6-carboxypyridin-2-yl)methyl)-1,4,7-triazacyclononane) ligands based on the 1,4,7-triazacyclononane anchor were prepared and their lanthanide complexes were characterized by NMR, fluorescence and potentiometric studies. The [Gd(ebpatcn)(H(2)O)] complex displays a relaxivity of r(1) = 4.68 mM(-1) s(-1) at 45 MHz and 298 K, whereas r(1) = 4.55 mM(-1) s(-1) was measured for [Gd(Hpbpatcn)(H(2)O)] under the same conditions. The modified scaffold of the ligands with respect to the previously reported H(3)bpatcn (1-(carboxymethyl)-4,7-bis[(6-carboxypyridin-2-yl)methyl]-1,4,7-triazacyclononane) leads to an optimization of the properties of these gadolinium complexes. The replacement of an acetate binding group of the H(3)bpatcn ligand with a propionate group (H(3)ebpatcn) or a phosphonate group (H(4)pbpatcn) leads to a faster exchange rate of the coordinated water molecule in both mono-aquo gadolinium complexes. The resulting water exchange rate is optimized for the future design of high relaxivity macromolecular gadolinium based contrast agents with a value measured by O(17) NMRD of k(ex) = 34 x 10(6) s(-1) for [Gd(Hpbpatcn)(H(2)O)] falling in the range of optimum values of (30 to 50) x 10(6) s(-1) predicted by the SBM theory. The water exchange rate k(ex)(298) = 86 x 10(6) s(-1) of the complex [Gd(ebpatcn)(H(2)O)] is the fastest reported in the literature for a neutral complex with only one inner-sphere water molecule. The relatively high stability of these modified gadolinium complexes (pGd = 14.1 for Gd(pbpatcn) and 13.1 for Gd(ebpatcn)) is similar to that of the [Gd(bpatcn)(H(2)O)] complex (pGd = 13.6). The high luminescence efficiency is also retained for the terbium complex. However, whereas the longitudinal electronic spin relaxation time keeps a value for [Gd(ebpatcn)(H(2)O)], which is long enough not to affect the relaxivity in macromolecular complexes (transient ZFS amplitude Delta(2) [10(20) rad(2) s(-2)] = 0.39), the O(17) relaxation and the (1)H NMRD indicate a rather fast electron spin relaxation for the phosphonate containing complex (Delta(2) [10(20) rad(2) s(-2)] = 1.3).


Mechanotransduction in cardiac hypertrophy and failure.
Cardiac muscle cells have an intrinsic ability to sense and respond to mechanical load through a process known as mechanotransduction. In the heart, this process involves the conversion of mechanical stimuli into biochemical events that induce changes in myocardial structure and function. Mechanotransduction and its downstream effects function initially as adaptive responses that serve as compensatory mechanisms during adaptation to the initial load. However, under prolonged and abnormal loading conditions, the remodeling processes can become maladaptive, leading to altered physiological function and the development of pathological cardiac hypertrophy and heart failure. Although the mechanisms underlying mechanotransduction are far from being fully elucidated, human and mouse genetic studies have highlighted various cytoskeletal and sarcolemmal structures in cardiac myocytes as the likely candidates for load transducers, based on their link to signaling molecules and architectural components important in disease pathogenesis. In this review, we summarize recent developments that have uncovered specific protein complexes linked to mechanotransduction and mechanotransmission within the sarcomere, the intercalated disc, and at the sarcolemma. The protein structures acting as mechanotransducers are the first step in the process that drives physiological and pathological cardiac hypertrophy and remodeling, as well as the transition to heart failure, and may provide better insights into mechanisms driving mechanotransduction-based diseases.


Structure of the aflatoxin B(1) dialdehyde adduct formed from reaction with methylamine.
Amine conjugates of activated aflatoxin (AF) B(1) are formed in various systems, e.g., buffer amines and with protein lysine groups. Structures have been published in the literature, but the evidence is indirect in that (i) halogenated AFB(1) was usually used as the precursor and (ii) the assignment of the structure of the five-membered ring formed by cyclization is based on NMR chemical shifts. To better define these adducts and distinguish among several possibilities, we synthesized AFB(1) dialdehyde and reacted this with the surrogate methylamine at neutral pH, to simplify the system. The isolated product had the expected molecular ion (mass spectrometry) and showed pH-dependent UV spectra similar to those published for a lysine conjugate. Nuclear Overhauser enhanced spectroscopy (two-dimensional NMR, 800 MHz) of the sample (2H(2)O) showed proximity of the N-CH(3) protons only with a singlet at delta 4.10, assigned to the methylene of the added five-membered ring, but not to a delta 6.53 singlet assigned as the vinylic proton of that ring. All protons in the coumarin-furanone portion of the system were correlated to each other but not to those in the added five-membered ring. These experiments establish the structure as 2,3-dihydro-2-oxo-4-(1,2,3,4-tetrahydro-7-hydroxy-9-methoxy-3,4-dioxocyclopenta[c][1]benzopyran-6-yl)-1H-pyrrole-1-methane. The similarity of the reaction to that occurring in the reaction of AFB(1) dialdehyde with lysine and the agreement of the UV spectra suggest that this structure is applicable for the lysine analogue. The NMR results support the possible structure B of Sabbioni et al. [Sabbioni, G., Skipper, P. L., Buchi, G., and Tannenbaum, S. R. (1987) Carcinogenesis 8, 819-824] and the proposed structure 8 of Sabbioni [Sabbioni, G. (1990) Chem.-Biol. Interact. 75, 1-15] but not alternative proposals. Kinetic and mechanistic considerations of the reaction of lysine with AFB(1) dialdehyde are presented in the previous article in this issue [Guengerich, F. P., Arneson, K. O., Williams, K. M., Deng, Z., and Harris, T. M. (2002) Chem. Res. Toxicol. 15, 780-793].


Modification of properties of yttria stabilized zirconia epitaxial thin films by excimer laser annealing.
This study focuses on the ultrafast improvement of surface wettability, electrical, and room temperature magnetic characteristics of cubic zirconia single crystalline thin films after laser annealing. The point defects generated by the laser treatment are envisaged to play a critical role in altering the above properties. Yttria stabilized zirconia (YSZ) thin films were epitaxially grown on Si(100) substrates by pulsed laser deposition technique and subsequently annealed by a KrF excimer laser beam (τ = 25 ns) using low-energy laser pulses. An atomically sharp interface, parallel to the film free surface, between laser annealed layer and the pristine region was observed. The single crystalline nature of thin films was preserved following the laser treatment. The laser-solid interaction with YSZ led to the introduction of point defects, i.e., oxygen vacancies, resulting in a strained structure which, in turn, resulted in the formation of a tetragonal-like zirconia. With the increase of number of laser pulses the laser treated films got highly disordered due to the high concentration of the point defects, while maintaining their crystalline nature. Although the surface of the pristine sample showed weak hydrophilic characteristics (contact angle ∼ 73°), the laser annealed samples exhibited significantly improved hydrophilic characteristics. It was found that there is an optimum number of laser pulses where the maximum hydrophilicity (contact angle ∼ 22°) is obtained. The carrier concentration in the sample with the highest hydrophilicity was determined to be higher by about 5 orders of magnitude compared to the pristine sample. This sample possessed the lowest electrical resistivity. The laser annealed YSZ epilayers showed a superior room-temperature ferromagnetic behavior, compared to the pristine samples. A 2-fold enhancement in the magnetization of the samples was observed following the laser treatment which is a clear demonstration of the key role of defects and their transient distribution throughout the lattice. All these observations were correlated with the formation of point defects due to the photon interaction with YSZ and absorption of energy of the KrF laser photons to produce defects.


Candy consumption in childhood is not predictive of weight, adiposity measures or cardiovascular risk factors in young adults: the Bogalusa Heart Study.
There are limited data available on the longitudinal relationship between candy consumption by children on weight and other cardiovascular risk factors (CVRF) in young adults. The present study investigated whether candy consumption in children was predictive of weight and CVRF in young adults. A longitudinal sample of children 10 years (n = 355; 61% females; 71% European-Americans, 29% African-Americans) who participated in cross-sectional surveys from 1973 to 1984 (baseline) and in one of two surveys (follow-ups) as young adults [19-38 years; mean (SD) = 23.6 (2.6) years] in Bogalusa, LA, were studied. Dietary data were collected using 24-h dietary recalls at baseline and at one follow-up survey; a food frequency questionnaire was used in the other follow-up survey. Candy consumers were those consuming any amount of candy. Candy consumption was calculated (g day(-1) ) from baseline 24-h dietary recalls, and was used as a covariate in the adjusted linear mixed models. Dependent variables included body mass index (BMI) and CVRF measured in young adults. At baseline, 92% of children reported consuming candy [46 (45) g day(-1)]; the percentage decreased to 67% [20 (30) g day(-1)] at follow-up. No longitudinal relationship was shown between baseline candy consumption and BMI or CVRF in young adults, suggesting that candy consumption was not predictive of health risks later in life. The consumption of nutrient rich foods consistent with dietary recommendations is important, although modest amounts of candy can be added to the diet without potential adverse long-term consequences to weight or CVRF. Additional studies are needed to confirm these results.


The key hypoxia regulated gene CAIX is upregulated in basal-like breast tumours and is associated with resistance to chemotherapy.
Basal-like tumours account for 15% of invasive breast carcinomas and are associated with a poorer prognosis and resistance to therapy. We hypothesised that this aggressive phenotype is because of an intrinsically elevated hypoxic response. Microarrayed tumours from 188 patients were stained for hypoxia-inducible factor (HIF)-1alpha, prolyl hydroxylase (PHD)1, PHD2, PHD3 and factor inhibiting HIF (FIH)-1, and carbonic anhydrase (CA) IX stained in 456 breast tumours. Tumour subtypes were correlated with standard clincopathological parameters as well as hypoxic markers. Out of 456 tumours 62 (14%) tumours were basal-like. These tumours were positively correlated with high tumour grade (P<0.001) and were associated with a significantly worse disease-free survival compared with luminal tumours (P<0.001). Fifty percent of basal-like tumours expressed HIF-1alpha, and more than half expressed at least one of the PHD enzymes and FIH-1. Basal-like tumours were nine times more likely to be associated with CAIX expression (P<0.001) in a multivariate analysis. Carbonic anhydrase IX expression was positively correlated with tumour size (P=0.005), tumour grade (P<0.001) and oestrogen receptor (ER) negativity (P<0.001). Patients with any CAIX-positive breast tumour phenotype and in the basal tumour group had a significantly worse prognosis than CAIX-negative tumours when treated with chemotherapy (P<0.001 and P=0.03, respectively). The association between basal phenotype and CAIX suggests that the more aggressive behaviour of these tumours is partly due to an enhanced hypoxic response. Further, the association with chemoresistance in CAIX-positive breast tumours and basal-like tumours in particular raises the possibility that targeted therapy against HIF pathway or downstream genes such as CAs may be an approach to investigate for these patients.


Bending nematic liquid crystal membranes with phospholipids.
The interactions of phospholipids with liquid crystals have formed the basis for attractive biosensor technologies, but many questions remain concerning the basic physics and chemistry of these interactions. Phospholipids such as 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC), at sufficiently high (∼1 μM) concentrations and/or sufficiently long times, turn the liquid crystal director perpendicular to the LC/water interface. If the other side of the LC film is in contact with a surface that prefers perpendicular alignment, the LC film appears completely dark between crossed polarizers. Recently, however, Popov et al. (J. Mater. Chem. B, 2017, 5, 5061) noted that at even higher (∼10 μM) DLPC concentrations, the liquid crystal texture brightens again between crossed polarizers. To explain this surprising observation, it was suggested that the LC interface might bend. In this paper we show by optical surface profiler measurements that indeed the interface of the LC film of 4-cyano-4'-octylbiphenyl (8CB) suspended in a transmission electron microscopy (TEM) grid with openings of ∼0.5 mm in diameter bends towards the lipid-coated interface. We demonstrate that where the bending occurs, the bent interface exhibits extreme sensitivity to air pressure variations, producing an optical response with acoustic stimulation. Finally, we suggest a physical mechanism for this astonishing result.


Disease progression in young patients with COPD: rethinking the Fletcher and Peto model.
Chronic obstructive pulmonary disease (COPD), although frequent in older individuals, can also occur at younger age; this latter population has not been well described. We reviewed the functional progression of 1708 patients with COPD attending pulmonary clinics. Those with three or more annual spirometries were divided into those who, at enrolment, were ≤ 55 (n = 103) or ≥ 65 (n = 463) years of age (younger and older COPD, respectively). Baseline and annual changes in lung function (forced expiratory volume in 1 s (FEV1)) and BODE (body mass index, airflow obstruction, dyspnoea, exercise capacity) score were recorded and compared between both groups. Severity distribution by Global Initiative for Chronic Obstructive Lung Disease and BODE scores were similar in both groups, except for mild obstruction, which was higher in the younger group. Mean FEV1 decline was 38.8 and 40.6 mL · year(-1), while BODE scores increased 0.19 and 0.23 units per year, for younger and older COPD, respectively. Both groups had similar proportion of FEV1 rapid decliners (42% and 46%, respectively). The severity distribution and progression of disease in younger patients with COPD is similar to that of patients of older age. This observation suggests that younger individuals presenting with COPD develop the disease from an already compromised pulmonary and systemic status, complementing the model of steeper decline of lung function proposed by Fletcher and Peto.


Evaluation of the feeding value of Dichrostachys cinerea pods for fattening pigs in Cuba.
Dichrostachys cinerea (L.) Wight & Arn. is a tropical leguminous shrub widely regarded as an invasive species in Cuba, after having invaded a significant proportion of its arable land during the past decades. Concurrently, smallholder pig producers are highly constrained by the scarcity of protein feeds. This study aimed to assess the feeding value of D. cinerea pod meal (DCPM) as an alternative protein supplement for pigs in Cuban smallholder production systems. An on-farm feeding trial was carried out with three groups (N = 10) of growing-fattening pigs over 60 days, where DCPM replaced 0, 15, and 30% in DM of a dietary commercial concentrate. Then, in an in vivo digestibility trial with eight growing pigs, apparent digestibilities of DCPM were determined for dry matter (DM), organic matter (OM) and crude protein (CP). Finally, in vitro digestibilities for OM (fecal and ileal) and CP (ileal) were determined. In the feeding trial, pig body weight gains were not affected by increased dietary substitution levels of concentrate for DCPM. Blood parameters, with a few exceptions, did not show significant differences among groups. Values for in vivo OM and CP digestibilities were 40.81 and 50.26%, and substantially higher than in vitro values. In conclusion, our results showed that at least 30% of DM in commercial concentrate could be substituted by DCPM without affecting pig growth performances under Cuban smallholder conditions. The low digestibility of DCPM is, however, not acceptable for intensive pig production systems. In vitro enzyme digestibility methods developed for commercial pig feeds are not suitable for DCPM without further calibration.


Evolutionary time-scale of the begomoviruses: evidence from integrated sequences in the Nicotiana genome.
Despite having single stranded DNA genomes that are replicated by host DNA polymerases, viruses in the family Geminiviridae are apparently evolving as rapidly as some RNA viruses. The observed substitution rates of geminiviruses in the genera Begomovirus and Mastrevirus are so high that the entire family could conceivably have originated less than a million years ago (MYA). However, the existence of geminivirus related DNA (GRD) integrated within the genomes of various Nicotiana species suggests that the geminiviruses probably originated >10 MYA. Some have even suggested that a distinct New-World (NW) lineage of begomoviruses may have arisen following the separation by continental drift of African and American proto-begomoviruses ∼110 MYA. We evaluate these various geminivirus origin hypotheses using Bayesian coalescent-based approaches to date firstly the Nicotiana GRD integration events, and then the divergence of the NW and Old-World (OW) begomoviruses. Besides rejecting the possibility of a<2 MYA OW-NW begomovirus split, we could also discount that it may have occurred concomitantly with the breakup of Gondwanaland 110 MYA. Although we could only confidently narrow the date of the split down to between 2 and 80 MYA, the most plausible (and best supported) date for the split is between 20 and 30 MYA--a time when global cooling ended the dispersal of temperate species between Asia and North America via the Beringian land bridge.


Securinine, a GABAA receptor antagonist, enhances macrophage clearance of phase II C. burnetii: comparison with TLR agonists.
Innate immune cell stimulation represents a complementary approach to vaccines and antimicrobial drugs to counter infectious disease. We have used assays of macrophage activation and in vitro and in vivo phase II Coxiella burnetii infection models to compare and contrast the activity of a novel innate immune cell agonist, securinine, with known TLR agonists. As expected, TLR agonists, such as LPS (TLR4) and fibroblast-stimulating lipopeptide-1 (FSL-1; TLR2), induced macrophage activation and increased macrophage killing of phase II C. burnetii in vitro. FSL-1 also induced accelerated killing of C. burnetii in vivo. Securinine, a gamma-aminobutyric acid type A receptor antagonist, was found to induce TLR-independent macrophage activation in vitro, leading to IL-8 secretion, L-selectin down-regulation, and CD11b and MHC Class II antigen up-regulation. As seen with the TLR agonists, securinine also induced accelerated macrophage killing of C. burnetii in vitro and in vivo. In summary, as predicted by the literature, TLR agonists enhance macrophage killing of phase II C. burnetii in vitro, and at least for TLR2 agonists, this activity occurs in vivo as well. Securinine represents a novel macrophage agonist, which has similar effects as TLR agonists in this model yet apparently, does not act through known TLRs. Securinine has minimal toxicity in vivo, suggesting it or structurally similar compounds may represent novel, therapeutic adjuvants, which increase resistance to intracellular pathogens.


Water permeation across artificial I-quartet membrane channels: from structure to disorder.
Artificial water channels (AWCs) have been designed for water transport across membranes with the aim to mimic the high water permeability observed for biological systems such as aquaporins (∼108-109 water molecules per s per channel), as well as their selectivity to reject ion permeation at the same time. Recent works on designed self-assembling alkylureido-ethylimidazole compounds forming imidazole-quartet channels (I-quartets), have shown both high water permeability and total ionic-rejection. I-quartets are thus promising candidates for further development of AWCs. However, the molecular mechanism of water permeation as well as I-quartet organization and stability in a membrane environment need to be fully understood to guide their optimal design. Here, we use a wide range of all-atom molecular dynamics (MD) simulations and their analysis to understand the structure/activity relationships of the I-quartet channels. Four different types with varying alkyl chain length or chirality have been studied in a complex fully hydrated lipid bilayer environment at both microsecond and nanosecond scale. Microsecond simulations show two distinct behaviors; (i) two out of four systems maintain chiral dipolar oriented water wires, but also undergo a strong reorganization of the crystal shape, (ii) the two other I-quartet channels completely lose the initial organization, nonetheless keeping a water transport activity. Short MD simulations with higher time resolution were conducted to characterize the dynamic properties of water molecules in these model channels and provided a detailed hypothesis on the molecular mechanism of water permeation. The ordered confined water was characterized with quantitative measures of hydrogen-bond life-time and single particle dynamics, showing variability among I-quartet channels. We will further discuss the underlying assumptions, currently based on self-aggregation simulations and crystal patches embedded in lipid bilayer simulations and attempt to describe possible alternative approaches to computationally capture the water permeation mechanism and the self-assembly process of these AWCs.


Effect of environmental enrichment on escalation of cocaine self-administration in rats.
Previous studies found that environmental enrichment protects against the initiation of stimulant self-administration in rats, but it is unclear if enrichment also protects against the escalation of stimulant use with long-term exposure. The current study examined the effects of environmental enrichment on escalation of cocaine self-administration using an extended access procedure. Rats were raised from 21 days in an enriched condition (EC) with social cohorts and novel objects, a social condition with only social cohorts (SC), a novelty condition (NC) with novel objects in isolated cages, or an isolated condition (IC) without social cohorts or novel objects. In young adulthood, EC, SC, NC, and IC rats were separated into short access (ShA) or long access (LgA) groups that received either 1 or 6 h, respectively, of daily cocaine self-administration (0.1 mg/kg/infusion) for 14 days. In a second experiment, EC and IC rats were used to assess differences in acquisition and escalation of cocaine self-administration at a 0.5 mg/kg/infusion unit dose. With ShA sessions, EC rats acquired cocaine self-administration at a slower rate than IC rats at both unit doses; however, with extended training, both groups eventually reached similar rates. At the 0.1 mg/kg/infusion dose, only NC and IC rats escalated in amount of intake when switched to the LgA sessions. At the 0.5 mg/kg/infusion dose, rates of cocaine self-administration escalated in LgA groups over 14 days regardless of EC or IC rearing condition; however, EC rats escalated at a faster rate, eventually reaching the same level of intake observed in IC rats. Although environmental enrichment protects against escalation of a low unit dose of cocaine, it may not protect against escalation with a higher unit dose. In addition, at a lower unit dose, this protective mechanism appears to be due to the presence of social cohorts rather than novel objects.


Synergistic effects of piperlongumine and gemcitabine against KRAS mutant lung cancer.
To determine the combined efficacy of piperlongumine and gemcitabine for treatment of KRAS mutant lung cancer. The cell growth inhibition of piperlongumine, gemcitabine, and piperlongumine plus gemcitabine was measured by Cell Counting Kit‑8 assay and the combination index was calculated. In addition, the combined effects of piperlongumine and gemcitabine on cell apoptosis, reactive oxygen species (ROS) contents, and microtubule-associated protein 1 light chain 3B (LC3B) expression were examined. Piperlongumine increased ROS contents and LC3B-II expression. Following the combined treatment with piperlongumine and 10 mM N-acetyl-L-cysteine (NAC), intracellular ROS and cell viability returned to normal levels, and the expression of LC3B-II decreased to the predose level. Gemcitabine also induced cell apoptosis, increased ROS contents, and LC3B-II expression. The combination of piperlongumine with gemcitabine exhibited a synergetic anticancer activity with the combination index <1. The combined application of gemcitabine and piperlongumine yielded synergistic effects on cell apoptosis, but failed to synergistically increase ROS levels and LC3B-II expression. Combination therapy with piperlongumine and gemcitabine is a promising treatment option for KRAS mutant lung cancer.


Sequential design of phase II-III cancer trials.
Although traditional phase II cancer trials are usually single arm, with tumor response as endpoint, and phase III trials are randomized and incorporate interim analyses with progression-free survival or other failure time as endpoint, this paper proposes a new approach that seamlessly expands a randomized phase II study of response rate into a randomized phase III study of time to failure. This approach is based on advances in group sequential designs and joint modeling of the response rate and time to event. The joint modeling is reflected in the primary and secondary objectives of the trial, and the sequential design allows the trial to adapt to increase in information on response and survival patterns during the course of the trial and to stop early either for conclusive evidence on efficacy of the experimental treatment or for the futility in continuing the trial to demonstrate it, on the basis of the data collected so far.


Design and Implementation of a Visual Analytics Electronic Antibiogram within an Electronic Health Record System at a Tertiary Pediatric Hospital.
Hospitals use antibiograms to guide optimal empiric antibiotic therapy, reduce inappropriate antibiotic usage, and identify areas requiring intervention by antimicrobial stewardship programs. Creating a hospital antibiogram is a time-consuming manual process that is typically performed annually. We aimed to apply visual analytics software to electronic health record (EHR) data to build an automated, electronic antibiogram ("e-antibiogram") that adheres to national guidelines and contains filters for patient characteristics, thereby providing access to detailed, clinically relevant, and up-to-date antibiotic susceptibility data. We used visual analytics software to develop a secure, EHR-linked, condition- and patient-specific e-antibiogram that supplies susceptibility maps for organisms and antibiotics in a comprehensive report that is updated on a monthly basis. Antimicrobial susceptibility data were grouped into nine clinical scenarios according to the specimen source, hospital unit, and infection type. We implemented the e-antibiogram within the EHR system at Children's Hospital of Philadelphia, a tertiary pediatric hospital and analyzed e-antibiogram access sessions from March 2016 to March 2017. The e-antibiogram was implemented in the EHR with over 6,000 inpatient, 4,500 outpatient, and 3,900 emergency department isolates. The e-antibiogram provides access to rolling 12-month pathogen and susceptibility data that is updated on a monthly basis. E-antibiogram access sessions increased from an average of 261 sessions per month during the first 3 months of the study to 345 sessions per month during the final 3 months. An e-antibiogram that was built and is updated using EHR data and adheres to national guidelines is a feasible replacement for an annual, static, manually compiled antibiogram. Future research will examine the impact of the e-antibiogram on antibiotic prescribing patterns.


Genotyping and Identification of Antigen B Gene Polymorphism of Echinococcus granulosus in Edirne, Thrace, and the First Report of Genotype G2 (Tasmanian Sheep Strain) in Turkey
Echinococcus granulosus is the causative agent of cystic echinococcosis in humans and livestock. It is common worldwide. Cystic echinococcosis is still an important public health problem in Turkey, which is an endemic region. To genotype Echinococcus granulosus isolates and investigate antigen B gene polymorphism in Thrace, Turkey. A cross-sectional study. Seventy-five hydatid cyst materials obtained between June 2020 and May 2021 were included in the study. Hydatid cyst materials were collected from 12 humans from various hospitals in Edirne and 63 from slaughterhouse animals during the same period. Cyst materials were localized in 8 livers and 4 lungs in humans, 23 livers and 17 lungs in cattle, and 13 livers and 10 lungs in sheep. In the first step, the 12S ribosomal RNA gene was amplified by polymerase chain reaction for all samples and run on an agarose gel. Band patterns were used for strain typing. Then, the selected samples that represented each of the band patterns obtained by single-strand conformation polymorphism analysis were sequenced for AgB1, AgB2, mt-CO1, and mt-ND1 genes. Three different genotypes in Edirne, Thrace, Turkey, were observed for Echinococcus granulosus: G1 (domestic sheep strain), G2 (Tasmanian sheep strain), and G3 (buffalo strain). G1 was the dominant genotype in Edirne, and G3 was the second most common. Additionally, polymorphism in AgB1 and AgB2 gene regions was found. This study is the first to report on Echinococcus granulosus G2 (Tasmania sheep strain) in Turkey and G3 (buffalo strain) and antigen B polymorphism in Thrace. The study results will contribute to the prevention and control programs for cystic echinococcosis in Turkey and worldwide.