USP15 Participates in Hepatitis C Virus Propagation through Regulation of Viral RNA Translation and Lipid Droplet Formation.
Hepatitis C virus (HCV) utilizes cellular factors for efficient propagation. Ubiquitin is covalently conjugated to the substrate to alter its stability or to modulate signal transduction. In this study, we examined the importance of ubiquitination for HCV propagation. We found that inhibition of deubiquitinating enzymes (DUBs) or overexpression of nonspecific DUBs impaired HCV replication, suggesting that ubiquitination regulates HCV replication. To identify specific DUBs involved in HCV propagation, we set up RNA interference (RNAi) screening against DUBs and successfully identified ubiquitin-specific protease 15 (USP15) as a novel host factor for HCV propagation. Our studies showed that USP15 is involved in translation of HCV RNA and production of infectious HCV particles. In addition, deficiency of USP15 in human hepatic cell lines (Huh7 and Hep3B/miR-122 cells) but not in a nonhepatic cell line (293T cells) impaired HCV propagation, suggesting that USP15 participates in HCV propagation through the regulation of hepatocyte-specific functions. Moreover, we showed that loss of USP15 had no effect on innate immune responses in vitro and in vivo We also found that USP15-deficient Huh7 cells showed reductions in the amounts of lipid droplets (LDs), and the addition of palmitic acids restored the production of infectious HCV particles. Taken together, these data suggest that USP15 participates in HCV propagation by regulating the translation of HCV RNA and the formation of LDs.IMPORTANCE Although ubiquitination has been shown to play important roles in the HCV life cycle, the roles of deubiquitinating enzymes (DUBs), which cleave ubiquitin chains from their substrates, in HCV propagation have not been investigated. Here, we identified USP15 as a DUB regulating HCV propagation. USP15 showed no interaction with viral proteins and no participation in innate immune responses. Deficiency of USP15 in Huh7 cells resulted in suppression of the translation of HCV RNA and reduction in the amounts of lipid droplets, and the addition of fatty acids partially restored the production of infectious HCV particles. These data suggest that USP15 participates in HCV propagation in hepatic cells through the regulation of viral RNA translation and lipid metabolism.


The influence of feminist abortion accompaniment on emotions related to abortion: A longitudinal observational study in Mexico.
Emotions can reflect how individuals internalize identities, social roles, and broader power structures, including abortion stigma. Abortion accompaniment, in the form of logistical, informational, and emotional support offered by individuals and organizations, takes a person-centered, feminist, and rights-based approach. We tested the extent to which abortion accompaniment may decrease negative and increase positive feelings an individual holds related to their abortion. Using observational longitudinal data collected between January 2017 and mid-2018, we compared negative and positive emotional responses to a personal abortion experience one month and six months following the abortion to emotions immediately prior to the abortion ("baseline"), among women travelling from outside of Mexico City to abortion clinics in Mexico City with and without support of the accompaniment organization, Fondo MARIA. We used doubly robust longitudinal mixed effects models with inverse probability weighting methods. At baseline, accompanied and unaccompanied participants experienced an average of 4.9 and 4.4 negative emotions out of eight respectively (i.e., anguish, nervousness, scared, anxious, sadness, guilt, anger, shame) and 1.7 and 1.9 positive emotions of out 4 respectively (happiness, calmness, decidedness, and relief). From our model results, women accompanied (n = 77) had larger decreases in negative feelings (p < .05) and larger increases in positive feelings (p < .01) toward their abortion compared to those who were not accompanied (n = 119) at six months. These changes led the majority of accompanied respondents to have primarily positive feelings about their abortion by endline. Abortion accompaniment through Fondo MARIA in Mexico City was associated with a larger decrease in negative feelings, particularly those related to stigma, and a larger increase in positive feelings six months after abortion. Accompaniment's focus on person-centered support, self-determination, and autonomy may enable people seeking abortion to view their decision as one that is valid and legitimate, and resist the predominant stigmatizing narratives framing abortion as something that is transgressive.


Histidine nutrition and genotype affect cataract development in Atlantic salmon, Salmo salar L.
The aim of this study was to investigate effects of dietary levels of histidine (His) and iron (Fe) on cataract development in two strains of Atlantic salmon monitored through parr-smolt transformation. Three experimental diets were fed: (i) a control diet (CD) with 110 mg kg(-1) Fe and 11.7 g kg(-1) His; (ii) CD supplemented with crystalline His to a level of 18 g kg(-1) (HD); and (iii) HD with added iron up to 220 mg kg(-1) (HID). A cross-over design, with two feeding periods was used. A 6-week freshwater (FW) period was followed by a 20-week period, of which the first three were in FW and the following 17 weeks in sea water (SW). Fish were sampled for weighing, cataract assessment and tissue analysis at five time points. Cataracts developed in all groups in SW, but scores were lower in those fed high His diets (P < 0.05). This effect was most pronounced when HD or HID was given in SW, but was also observed when these diets were given in FW only. Histidine supplementation had a positive effect on growth performance and feed conversion ratio (P < 0.05), whereas this did not occur when iron was added. Groups fed HD or HID had higher lens levels of His and N-acetyl histidine (NAH), the latter showing a marked increase post-smoltification (P < 0.05). The HD or HID groups also showed higher muscle concentrations of the His dipeptide anserine (P < 0.05). There was a strong genetic influence on cataract development in the CD groups (P < 0.001), not associated with tissue levels of His or NAH. The role of His and His-related compounds in cataractogenesis is discussed in relation to tissue buffering, osmoregulation and antioxidation.


Disease-related blood-based differential methylation in cystic fibrosis and its representation in lung cancer revealed a regulatory locus in PKP3 in lung epithelial cells.
Cystic fibrosis (CF) is a monogenic disease, characterized by massive chronic lung inflammation. The observed variability in clinical phenotypes in monozygotic CF twins is likely associated with the extent of inflammation. This study sought to investigate inflammation-related aberrant DNA methylation in CF twins and to determine to what extent acquired methylation changes may be associated with lung cancer.Blood-based genome-wide DNA methylation analysis was performed to compare the DNA methylomes of monozygotic twins, from the European CF Twin and Sibling Study with various degrees of disease severity. Putatively inflammation-related and differentially methylated positions were selected from a large lung cancer case-control study and investigated in blood by targeted bisulphite next-generation-sequencing. An inflammation-related locus located in the Plakophilin-3 (PKP3) gene was functionally analysed regarding promoter and enhancer activity in presence and absence of methylation using luciferase reporter assays.We confirmed in a unique cohort that monozygotic twins, even if clinically discordant, have only minor differences in global DNA methylation patterns and blood cell composition. Further, we determined the most differentially methylated positions, a high proportion of which are blood cell-type-specific, whereas others may be acquired and thus have potential relevance in the context of inflammation as lung cancer risk factors. We identified a sequence in the gene body of PKP3 which is hypermethylated in blood from CF twins with severe phenotype and highly variably methylated in lung cancer patients and controls, independent of known clinical parameters, and showed that this region exhibits methylation-dependent promoter activity in lung epithelial cells.


Hyperacute cerebral fat embolism in a patient with femoral shaft fracture.
Fat embolism syndrome is a potentially fatal complication and occurs most commonly after long bone fracture. In patients who sustained severe trauma, both cerebral fat embolism(CFE) and diffuse axonal injury (DAI) could be the cause of altered consciousness in the absence of marked intracranial lesions in cranial computed tomography. However, distinguishing CFE and DAI can be difficult clinically. Generally, DAI develops immediately after the insult, whereas CFE occurs 48 to 72 hours after the trauma and even after internal fixation for the fractures. Fat embolism syndrome develops within an average of 48.5 hours after long bone fracture [1] but has never been reported to occur in less than 2 hours. Here, we present a patient who developed hyperacute CFE and eventually had poor neurological outcome, in contrast to previous reports stating that CFE usually has a long latent period and favorable outcomes.


Self-admission in psychiatry: The ethics.
Self-admission to inpatient treatment is a novel approach that aims to increase agency and autonomy for patients with severe psychiatric illness and a history of high utilization of inpatient care. By focusing on brief, preventive hospital admissions in times of increased risk of relapse, self-admission seeks to reduce the need for prolonged episodes of inpatient treatment. Participants are generally satisfied with the model, which is not surprising given that self-admission programs allocate a scarce resource-hospital beds-to a select group. However, the patients targeted by these programs are not necessarily those in most need of hospital admission, which may compromise the commonly accepted 'principle of need' in allocation of public healthcare resources. In light of the current lack of consistent evidence of the usefulness and cost-effectiveness of the model, several aspects need to be further studied in order to guide any large-scale implementation of self-admission in psychiatry.


Intraoperative sentinel node identification with Technetium-99m-labeled nanocolloid in patients with cancer of the uterine cervix: a feasibility study.
The objective of this study is to determine the feasibility of intraoperative lymphatic mapping in patients with cervical carcinoma. Patients with early-stage cervical cancer, scheduled to undergo a Wertheims radical hysterectomy and pelvic lymphadenectomy, were eligible for the study. Technetium-99-m-labeled nanocolloid was injected intracervically at two locations around the tumor 3-6 hours before the operation. Images were recorded immediately and 2.5 hours later using a gamma camera. During the operation sentinel nodes (SLN) were identified using a handheld or laparoscopic gamma-detection probe (Navigator, Auto-Suture). After resection of the SLNs a standard pelvic (and para-aortic) lymphadenectomy was performed. The results of the histopathology of SLNs and non-SLNs were compared. The procedure was well tolerated by 24 of 25 patients. One or more SLN could be identified in 21 out of 25 patients. In one patient who was preoperatively irradiated, in two patients who had had a cone biopsy, and in one patient without previous interventions, no SLN could be detected. The mean time for identification was 5 minutes. In 16 patients, pathologic examination showed no metastatic disease in both SLNs and non-SLNs, whereas metastases were found in the SLN in five patients (in one case only SLN involved, four cases SLN and respectively 3, 11, 22 and 31 other positive nodes). This study shows that identification of SLNs in cancer of the uterine cervix is feasible with preoperatively administered 99mTc-labeled nanocolloid. To date, no false negative SLNs have been found, but expansion of the study is necessary to determine possible clinical application of this new technique.


A complex mechanism of facilitation in pituitary ACTH cells: recent single-cell studies.
The transfer of information by chemical signals during complex biological processes can, with increasing frequency, be described in terms of interacting signal pairs. External signalling is rarely monolithic; rather, signal pairs are utilized in processes such as hormone secretion, neurotransmission, cell growth and differentiation. The dualism of external signalling often results in the occurrence of synergy. One signal appears to turn the cell on or off, and its synergistic partner increases cell responsiveness, providing gain control of the cellular response. ACTH release provoked by certain stressors arises from a synergistic interaction between two hypothalamic hormones: corticotropin releasing hormone (CRH) and a modulator such as vasopressin (AVP). The pituitary ACTH cell has been used to unravel the intracellular messenger equivalents of an external signal interaction that generates synergy. Research emphasizes the single cell approach. Direct measurements of intracellular free Ca2+ were performed using the Ca2+-sensitive fluorescent probe fura-2/AM and instrumentation for digital image processing. A reverse haemolytic plaque assay was used to measure cumulative ACTH release from single pituitary cells in culture. What is the physiological role of intracellular Ca2+ as a messenger? What are the feedforward and feedback relationships between major second messengers [cyclic AMP, diacylglycerol (DAG), inositol 1,4,5-trisphosphate (Ins(1,4,5)P3)] and intracellular Ca2+? That is, how are individual messenger circuits "wired' within ACTH cells. Intracellular Ca2+ may act as a common signal into which interacting second messenger signals [cyclic AMP, Ins(1,4,5)P3, DAG] are transduced and integrated to govern ACTH release. A novel circuit of messenger pathways linked by Ca2+ is proposed as the intracellular basis for the synergistic interaction of CRH- and AVP-regulated ACTH release.


Diffusion and binding measurements within oral biofilms using fluorescence photobleaching recovery methods.
Numerous studies have postulated that bacteria which reside in a biofilm differ from planktonic bacteria. These differences are thought to affect biofilm permeability and, indirectly, the susceptibility of biofilm bacteria to antibacterial agents. In this study fluorescence recovery after photobleaching (FRAP) was used to monitor the diffusion and binding characteristics of a set of size fractionated fluorescein isothiocyanate (FTTC)-conjugated dextrans over small areas (ca. 10 micron) in bacterial biofilms. From these measurements it was straightforward to calculate apparent diffusion rates. Initial studies on the concentration dependence of dextran interaction with planktonic bacteria showed that no irreversible interaction was occurring, however, anomalous faster than free solution diffusion rates were obtained. This phenomenon was modelled using novel analytical and numerical methods which incorporate reversible binding with associated fluorescence changes. Apparent diffusion rates measured in biofilms were highly dependent on biofilm preparation. Sucrose starved biofilms produced an apparent slow-down of two- to fivefold depending on dextran molecular mass and location within the biofilm, indicating that diffusion within the biofilm is hindered. Sucrose supplemented biofilms produced apparent diffusion rates close to those in free solution, suggesting less hindered diffusion. Ex vivo plaque showed diffusion and binding similar to the sucrose supplemented biofilms. The FRAP approach provides a fast and convenient method for determining diffusion rates over small areas within bacterial biofilms. This study reinforces the importance of considering the influence of reversible binding and associated fluorescence changes, as these may have a marked effect on the measured apparent diffusion rate.


[Rupture of the uterine artery after spontaneous delivery with unusual course in childbed].
Haemorrhage from ruptured uterine vessels is a rare but life-threatening complication during pregnancy. The high rate of mortality associated with this condition is correlated to the rapidity of haematoma formation. The dynamics of this event become particularly evident at the time of birth when the uterus requires one-fifth of the cardiac output. This high rate of flow emphasizes the possible fatal consequences of delayed therapeutical intervention. Information available on the aetiology and pathogenesis of such events is poor, especially if an involvement of cervical vessels or extraperitoneal (incomplete) rupture of the uterus can be excluded. Diagnosis and therapy are based on the classical clinical symptoms of acute abdominal pain-and-shock symptomatology. We report on a 31-year old patient after spontaneous delivery and initially uneventful puerperium. She had to be laparotomised on the third day post partum because of pain and haemoglobin deficiency. We found a large retroperitoneal (supralevatorial) haematoma caused by ruptured lateral branches of the left arteria uterina. The uterus was preserved after haematoma removal and revision and suture of the bleeding vessels. The case reported on shows that peripartal rupture of arterial uterine vessels may turn out to be clinically relevant only after a relatively long time (> 72 hours). Only four comparable cases have been reported in the literature, the course being different from that in all the other cases. In all the documented cases arterial uterine haemorrhages resulted in short-term acute aggravation of the symptomatology with severe abdominal pain, haemodynamic collapse and simultaneous haemoglobin deficit. The case under report in the present paper was at first clinically occult and eventually developed a pathology that had not been described before. Its specific significance lies in illustrating the need for re-evaluating such initially hidden cases while paying special attention to ruptured uterine vessels.


Mutational studies with Atp12p, a protein required for assembly of the mitochondrial F1-ATPase in yeast. Identification of domains important for Atp12p function and oligomerization.
The Atp12p protein of Saccharomyces cerevisiae is required for assembly of the F1 moiety of the mitochondrial ATP synthase. The current work has used mutant forms of Atp12p in an effort to learn about amino acids and/or domains that are important for the action of the protein. In one set of studies, the mutant atp12 genes were cloned and sequenced from 13 independent isolates of chemically mutagenized yeast. Of the 10 different mutant alleles that were identified, 9 (8 nonsense and 1 frameshift) lead to the early termination of the protein. A single missense mutation that substitutes lysine for Glu-289 was identified in two of the atp12 strains. Analysis of several Atp12p variants, each with different substitutions at Glu-289, showed that the functional activity of Atp12p is compromised when non-acidic residues are introduced at position 289 in the sequence. In other work, deletion analysis led to the assignment of two domains in Atp12p; the functional domain of the protein was mapped to the sequence between Gln-181 and Val-306, and a structural domain (Asp-307 through Gln-325) was identified that confers Atp12p the ability to oligomerize with other proteins in mitochondria.


Two-year outcome with Nobel Direct implants: a retrospective radiographic and microbiologic study in 10 patients.
The Nobel Direct implant (Nobel Biocare AB, Göteborg, Sweden) was developed to minimize marginal bone resorption and to result in "soft tissue integration" for an optimized aesthetic outcome. However, conflicting results have been presented in the literature. The aim of this present study was to evaluate the clinical and microbiologic outcomes of Nobel Direct implants. Ten partially edentulous subjects without evidence of active periodontitis (mean age 55 years) received 12 Nobel Direct implants. Implants were loaded with single crowns after a healing period of 3 to 6 months. Treatment outcomes were assessed at month 24. Routine clinical assessments, intraoral radiographs, and microbiologic samplings were made. Histologic analysis of one failing implant and chemical spectroscopy around three unused implants was performed. Paired Wilcoxon signed-rank test was used for the evaluation of bone loss; otherwise, descriptive analysis was performed. Implants were functionally loaded after 3 to 6 months. At 2 years, the mean bone loss of remaining implants was 2.0 mm (SD +/- 1.1 mm; range: 0.0-3.4 mm). Three out of 12 implants with an early mean bone loss >3 mm were lost. The surviving implants showed increasing bone loss between 6 and 24 months (p = .028). Only 3 out of the 12 implants were considered successful and showed bone loss of <1.7 mm after 2 years. High rates of pathogens, including Aggregatibacter actinomycetemcomitans, Fusobacterium spp., Porphyromonas gingivalis, Pseudomonas aeruginosa, and Tanerella forsythia, were found. Chemical spectroscopy revealed, despite the normal signals from Ti, O, and C, also peaks of P, F, S, N, and Ca. A normal histologic image of osseointegration was observed in the apical part of the retrieved implant. Radiographic evidence and 25% implant failures are indications of a low success rate. High counts and prevalence of significant pathogens were found at surviving implants. Although extensive bone loss had occurred in the coronal part, the apical portion of the implant showed some bone to implant integration.


The bone morphogenetic protein 15 gene is X-linked and expressed in oocytes.
We have taken advantage of the sequence relationships among the bone morphogenetic proteins (BMPs) to identify the mouse Bmp15 and human BMP15 genes. The 392-amino acid prepropeptides encoded by these BMP genes exhibit significant homology to each other, although the 70% identity observed between the 125-amino acid mature peptides is considerably lower than that seen in comparisons of other mouse and human orthologs. Both genes share a common structural organization and encode mature peptides that lack the cysteine residue normally involved in the formation of a covalent dimer. In addition, mouse Bmp15 and human BMP15 map to conserved syntenic regions on the X chromosome. We demonstrate, through a combination of Northern blot and in situ hybridization analyses, that mouse Bmp15 is expressed specifically in the oocyte beginning at the one-layer primary follicle stage and continuing through ovulation. Interestingly, BMP-15 is most closely related to and shares a coincident expression pattern with the mouse growth/differentiation factor 9 (GDF-9) gene that is essential for female fertility. Our findings will be important for defining the role of BMP-15 in follicular development.


The effects of domain deletion, glycosylation, and long IgG3 hinge on the biodistribution and serum stability properties of a humanized IgG1 immunoglobulin, hLL2, and its fragments.
Antibody (Ab) fragments are preferred agents for imaging applications because of their rapid clearance from the blood, thereby providing high tumor:blood ratios within a few hours. Several preclinical studies have also suggested that Ab fragments might be preferred for therapeutic applications over an intact IgG. The purpose of this project was to develop engineered Ab fragments using a humanized anti-carcinoembryonic antigen and anti-CD22 Ab as the parent. Three types of variants were prepared: a deltaCH2 (deletion mutant missing the CH2), a gamma3 F(ab')2 containing the human IgG3 hinge, and three glycosylated variants. The gamma3 F(ab')2 and glycosylated variants were developed because of the potential for site-specific linkage to the Ab in its divalent or monovalent fragment. The gamma3 F(ab')2 variant contains 10 cysteine residues that could be used for direct coupling using thiol chemistry, whereas the glycosylated variants have N-linked glycosylation sites engineered in the CH1 domain (two variants) as well as the VK domain (one variant). All of these variants were successfully prepared and shown to react with the target antigen. All Abs could be purified to a single peak by size-exclusion HPLC, but the deltaCH2 variant showed two distinct peaks, which were believed to be both the divalent and monovalent forms of this fragment. The two CH1 glycosylated variants showed differences in the extent of glycosylation. Modeling studies suggest that one variant would be better suited for site-specific coupling than the other because the carbohydrate chain is extended further away from the antigen-binding site. The Abs were radioiodinated to determine their pharmacokinetic behavior in mice. All of the humanized Ab divalent fragments cleared nearly 20 times faster from the blood than the murine parent F(ab')2 over a 24-h period. The glycosylated fragments showed some added stability compared to the other fragments over 4 h, but by 24 h, they had cleared to the same extent. Size-exclusion high-performance liquid chromatography of blood samples indicated that the humanized Ab fragments were quickly degraded in the blood. Thus, there is an inherent instability of the divalent fragments from these humanized IgG1 constructs that may affect their utility in imaging or therapy applications.


Distinct lobes of Limulus ventral photoreceptors. II. Structure and ultrastructure.
The structure of Limulus ventral photoreceptors fixed in situ has been investigated using light and electron microscopy and computer-assisted reconstruction and planimetry. Photoreceptors occur singly and in clusters. All photoreceptors have two types of lobes. The rhabdomeral lobe (R lobe) appears to be specialized for light sensitivity, containing the rhabdomere, which completely covers its external surface and forms infoldings into the lobe. The structure of the external rhabdom differs from that within infoldings. The other main structures of the R lobe are the palisades along the rhabdom, multivesicular bodies, lamellar bodies, and mitochondria. The arhabdomeral lobe (A lobe) bears the axon and contains the nucleus, clusters of residual bodies, lamellar arrays of endoplasmic reticulum, masses of glycogen, lipid droplets, and Golgi complexes. The R lobe and A lobe are analogous to the outer and inner segments of vertebrae photoreceptors. In single photoreceptors A and R lobes are separated by an indentation filled with glial processes. Computer reconstructions of cell clusters reveal that each cell has both types of lobes and an axon. Most of the rhabdom is formed from abutting arrays of external rhabdom from the R lobes of different members of the cluster. Efferent fibers containing characteristic angular granules penetrate single cells and clusters in glial invaginations. The main, if not exclusive, target of the efferent fibers is the internal rhabdom.


Colorectal cancer in patients under age 50: a five-year experience.
to evaluate the clinical characteristics of patients with colorectal cancer under the age of 50 treated at a public hospital in Brasilia over 5 years. we conducted a longitudinal, retrospective study, with 184 patients undergoing surgical procedures at the Asa Norte Regional Hospital (HRAN), including those who underwent only biopsy, between January 2013 and January 2018. We divided the patients into two groups: under the age of 50 (n=39) and age equal to or greater than 50 years (n=145). We compared the groups as to age, sex, symptoms, time between symptom onset and diagnosis, family and personal history, tumor location, histopathological characteristics, applied surgical management, staging and mortality. the group of patients under the age of 50 had more individuals with stage III and IV (p=0.041), more frequent poorly differentiated tumors (10.25% versus 3.52%; p=0.153), and higher incidences of compromised surgical margins (p=0.368), angiolymphatic (p=0.07) and perineural (p=0.007) invasion, which denotes more advanced disease in this group of patients. the study showed the low effectiveness of population screening methods for colorectal cancer currently used in this population, given the high incidence of the disease and late diagnosis in both groups.


Stress vulnerability in male youth with Internet Gaming Disorder.
Internet Gaming Disorder [IGD] was introduced as new behavioral addiction in DSM-5 Section 3. Vulnerability to stress is a potential predisposing factor for IGD. Given a lack of preexisting empirical data, the study investigated differences in the psychological and neurobiological response to acute stress in patients with IGD. 24 young men (mean age 18.38 years; range 13-25 years) fulfilling DSM-5 criteria for IGD and 25 matched controls underwent the Trier Social Stress Test [TSST]. Participants provided hair samples for the analysis of basal hypothalamic-pituitary-adrenal [HPA] axis activity and clinical interviews were conducted to assess psychopathology. During the experiment, participants reported on their subjective stress experience and momentary affect, provided samples of salivary cortisol and their heart rate was continuously recorded. Patients with IGD reported greater everyday and chronic stress, as well as psychopathological comorbidity. No differences were found on measures of hair cortisol. Compared to controls, IGD patients showed an attenuated cortisol response (χ2(7)=25.75, p<0.001) and greater negative affect (χ2(7)=17.25, p=0.016) in response to acute stress. Heart rate (χ2(1)=5.49, p=0.019), negative affect (χ2(1)=5.60, p=0.018) and subjective stress (χ2(1)=5.55, p=0.019) were transiently increased in IGD patients. After adjusting for sportive activities, IGD patients showed transiently decreased cortisol (χ2(1)=5.20, p=0.022), potentially indicating general HPA-axis dysfunction beyond altered reactivity. Stress reactivity showed correlations with IGD symptom severity. Findings illustrate differences in acute psychological and neurobiological stress reactivity in patients with IGD. Alterations of the stress response system may be involved in the development and maintenance of IGD.


Understanding Stakeholders' Perspectives to Increase COVID-19 Vaccine and Booster Uptake Among Black Individuals With Rheumatic Conditions.
Disparities in COVID-19 vaccine and booster uptake persist. This study aimed to obtain perspectives from community and physician stakeholders on COVID-19 vaccine and booster hesitancy and strategies to promote vaccine uptake among Black individuals with rheumatic and musculoskeletal conditions. We invited community leaders and physicians in greater Boston and Chicago to participate in semi-structured interviews using a moderator guide developed a priori. Participants were queried about how to best address vaccine hesitancy, strategies to target high-risk populations, and factors to identify future community leaders. Interviews were audio recorded, transcribed verbatim, and analyzed thematically using Dedoose. A total of 8 physicians and 12 community leaders participated in this study between November 2021 and October 2022. Qualitative analyses revealed misinformation/mixed messaging and mistrust, with subthemes including conspiracy theories, concerns regarding vaccine development and function, racism and historical injustices, and general mistrust of health care systems as the top cited reasons for COVID-19 vaccine hesitancy. Participants also shared demographic-specific differences, such as race, ethnicity, age, and gender that influenced the identified themes, with emphasis on COVID-19 vaccine access and apathy. Strategies for community-based vaccine-related information dissemination included personal storytelling with an iterative and empathetic approach, while recognizing the importance of protecting community leader well-being. To increase vaccine uptake among Black individuals with rheumatic conditions, strategies should acknowledge and respond to racial/ethnic and socioeconomic injustices that engender vaccine hesitancy. Messaging should be compassionate, individually tailored, and recognize heterogeneity in experiences and opinions. Results from these analyses will inform a planned community-based intervention in Boston and Chicago.


Fifteen Years of Airborne Particulates in Vitro Toxicology in Milano: Lessons and Perspectives Learned.
Air pollution is one of the world's leading environmental causes of death. The epidemiological relationship between outdoor air pollution and the onset of health diseases associated with death is now well established. Relevant toxicological proofs are now dissecting the molecular processes that cause inflammation, reactive species generation, and DNA damage. In addition, new data are pointing out the role of airborne particulates in the modulation of genes and microRNAs potentially involved in the onset of human diseases. In the present review we collect the relevant findings on airborne particulates of one of the biggest hot spots of air pollution in Europe (i.e., the Po Valley), in the largest urban area of this region, Milan. The different aerodynamic fractions are discussed separately with a specific focus on fine and ultrafine particles that are now the main focus of several studies. Results are compared with more recent international findings. Possible future perspectives of research are proposed to create a new discussion among scientists working on the toxicological effects of airborne particles.


Evidence on nutritional assessment techniques and parameters used to determine the nutritional status of children and adolescents: systematic review.
This article aims to review systematically the evidence on nutritional assessment techniques and parameters used to determine the nutritional status of children and adolescents. The literature review and the selection of publications were performed using the Medline, Lilacs, SciELO, Embase, personal files. 17 studies were identified, 7 addressed the anthropometric indices as the main outcome, 7 analyzed the growth and development of children and adolescents through growth curves, and the remainder surveyed body composition. In general, all met the quality criteria, unless 6 of the articles who did not discuss the limitations. The literature review suggests several techniques and parameters that can be applied to determine the nutritional status of children and adolescents from different countries. Growth graphs are essential to assess the health of children, but depend greatly of the growth tables used. Although BMI can be practical, it does not distinguish body fat from lean mass. The best interpretation of anthropometry will depend of valid reference values for age range of the study population. BIA is a quick feasible method, but the measurement has some various nationalities.


The insulin receptor substrate (IRS) proteins: at the intersection of metabolism and cancer.
Increasing evidence supports a connection between cancer and metabolism and emphasizes the need to understand how tumors respond to the metabolic microenvironment and how tumor cell metabolism is regulated. The insulin receptor (IR) and its close family member the insulin-like growth factor-1 receptor (IGF-1R) mediate the cellular response to insulin in normal cells and their function is tightly regulated to maintain metabolic homeostasis. These receptors are also expressed on tumor cells and their expression correlates with tumor progression and poor prognosis. Understanding how the IR/IGF-1R pathway functions in tumors is increasing in importance as the efficacy of drugs that target metabolic pathways, such as metformin, are investigated in prospective clinical trials. This review will focus on key signaling intermediates of the IR and IGF-1R, the Insulin Receptor Substrate (IRS) proteins, with an emphasis on IRS-2, and discuss how these adaptor proteins play a pivotal role at the intersection of metabolism and cancer.


Critical evaluation of in situ analyses for the characterisation of red pigments in rock paintings: A case study from El Castillo, Spain.
Paint technology, namely paint preparation and application procedures, is an important aspect of painting traditions. With the expansion of archaeometric studies and in situ non-destructive analytical methods, a renewal of technological studies is being observed in rock art. In situ analyses have several limitations that are widely discussed in the literature, however. It is not yet clear whether they provide accurate information on paint technology, except under certain conditions. Here, we evaluated digital microscopic and pXRF in situ analyses for the characterisation of a large set of red and yellow paintings from the El Castillo cave, Cantabria, Spain. We have set experiments and used statistical methods to identify differences between paint components and determine factors impacting pXRF measurements. We found that the compositional heterogeneity of the paintings' environment, especially variations in secondary deposits, was responsible for most of the differences observed between the pXRF signals recorded on the paintings. We concluded that the El Castillo cave environment is not suitable for non-destructive technological studies, but that more favourable contexts might exist. Following previous works and our own results, we advocate a combination of both in situ and laboratory invasive analyses for the study of paint composition and paint technology. Our research protocol, based on the comparison of rock paintings, their substrate, experimental paintings and Fe-normalisation of the signals can improve the reliability of pXRF results. We also propose to include more systematic characterisation of rock wall heterogeneity and the use of microscopic analyses in non-destructive approaches.


A fluorescence-based method to directly quantify antibodies immobilized on gold nanoparticles.
The ability to evaluate antibody immobilization onto gold nanoparticles is critical for assessing coupling chemistry and optimizing the sensitivity of nanoparticle-enabled biosensors. Herein, we developed a fluorescence-based method for directly quantifying antibodies bound onto gold nanoparticles. Antibody-modified gold nanoparticles were treated with KI/I2 etchant to dissolve the gold nanoparticles. A desalting spin column was used to recover the antibody released from the nanoparticles, and NanoOrange, a fluorescent dye, was used to quantify the antibody. We determined 309 ± 93 antibodies adsorb onto a 60 nm gold nanoparticles (2.6 × 10(10) NP mL(-1)), which is consistent with a fully adsorbed monolayer based on the footprint of an IgG molecule. Moreover, the increase in hydrodynamic diameter of the conjugated nanoparticle (76 nm) compared to that of the unconjugated nanoparticle (62 nm) confirmed that multilayers did not form. A more conventional method of indirectly quantifying the adsorbed antibody by analysis of the supernatant overestimated the antibody surface coverage (660 ± 87 antibodies per nanoparticle); thus we propose the method described herein as a more accurate alternative to the conventional approach.


Clinicopathologic Factors Predictive of Occult Lymph Node Involvement in Cutaneous Head and Neck Melanoma.
Objectives Evaluate clinicopathologic factors contributing to regional lymph node (LN) metastases in cutaneous head and neck melanoma (HNM). Study Design Retrospective review of a national cancer database. Setting National Cancer Database (2004-2012). Methods National Cancer Database (NCDB) cases diagnosed between 2004 and 2012 were reviewed. Inclusion criteria were head and neck cutaneous site codes, melanoma histology codes, and clinically node-negative status. Independent, clinicopathologic factors associated with pathologic positive LN status were identified by multivariable logistic regression. Subset analysis was performed on thin melanoma cases of 0.75 to 1 mm tumor depth to determine factors predictive of LN involvement and survival. Results Of 66,495 cases meeting inclusion criteria, 18,882 had nodes examined pathologically with 9.7% presenting with occult positive LN involvement. Mean (SD) age was 62.9 (16.13) years, and primary sites were scalp and neck (42.2%), face (40.2%), and external ear, lip, or eyelid (7.6%). Multivariable analysis found younger age; primary site of cutaneous scalp, neck, or face; increasing thickness; mitoses; ulceration; and vertical growth phase presence were independently associated with LN positivity ( P < .001). Only 2% of 0.75- to 1.0-mm Breslow thickness cases had regional LN involvement, and age >70 was negatively associated with LN involvement ( P = .002) in this subset. Conclusion This is the largest study examining factors predictive of occult LN involvement in patients with clinical node-negative cutaneous HNM who may benefit from sentinel LN biopsy. Primary sites of external ear, lip, or eyelid are less likely to present with occult nodal involvement, and older patients with T1a melanoma ≥0.76 mm are unlikely to present with regional node metastasis and may not require sentinel node guided management.


Resveratrol Partially Suppresses Inflammatory Events but Does not Affect Stroke Onset in Stroke-Prone Spontaneously Hypertensive Rats.
Resveratrol has been shown to mimic the beneficial effects of dietary restriction (DR). We previously reported that DR delays stroke onset and extends the lifespan in Stroke-Prone Spontaneously Hypertensive rats (SHRSP). Therefore, we examined whether resveratrol mimics DR and delays stroke onset in SHRSP. Cerebrovascular endothelial cells (CVECs) from SHRSP were treated with resveratrol, and the inflammatory gene expression levels and NFκB protein levels were measured. In order to address the effects of resveratrol in vivo, SHRSP (male, 10 weeks of age) were fed an experimental diet containing several doses of resveratrol (0 - 0.05% (w/w)), after which we measured the plasma cytokine levels and examined the stroke onset and lifespan. Treatment with resveratrol (100 μM, 24 hours) in CVECs from SHRSP significantly decreased the interleukin (IL)-1β-induced monocyte chemoattractant protein-1 (MCP-1) mRNA expression levels and p50 and p65 protein levels in the nuclear fraction. When the SHRSP were fed a diet containing resveratrol for one week, the resveratrol treatment did not affect the plasma lipid and glucose levels, body weight or weight of each tissue. Resveratrol slightly, but not significantly, decreased the plasma levels of IL-1β and MCP-1 compared with that observed in the control group. In addition, resveratrol decreased the IL-1β and MCP-1 mRNA expression levels in the brain versus the control animals. However, no doses of resveratrol delayed stroke onset or extended the lifespan in SHRSP. In this study, resveratrol did not delay stroke onset in SHRSP, although it partially suppressed systemic and cerebral inflammation. These results suggest that resveratrol does not mimic the beneficial effects of DR on stroke in vivo.


Increased oxidative stress in the mouse adriamycin model of glomerulosclerosis is accompanied by deposition of ferric iron and altered GGT activity in renal cortex.
Chronic renal failure evolves inevitable towards glomerular and tubulo-interstitial sclerosis. This pathological process involves a disturbed redox status of the kidney tissue, leading to irreversible damage. In this study we investigate in an adriamycin model of chronic renal failure in mice the evolution of in vivo hydrogen peroxide production, and the possible role of gamma-glutamyl transpeptidase and ferric iron in the process. Histological changes and ferric iron deposits are evaluated by histochemical staining. To evaluate oxidative stress residual catalase activity, TBARS formation and gamma-glutamyl transpeptidase activity are measured spectrophotometrically. While catalase activity remains the same, a decreased residual catalase activity indicates an increased formation of hydrogen peroxide. Both the activity of gamma-glutamyl transpeptidase and TBARS formation is increased at early stages of the disease. Ferric iron is clearly present in the proximal tubule. Twenty days after adriamycin injection all parameters decrease, probably due to the destruction of the tissue. Our data show the involvement of oxidative stress in the progression of adriamycin induced renal failure in mice. Both radical production and oxidative damage are measurable, while the altered activity of gamma-glutamyl transpeptidase and the deposition of ferric iron suggest the involvement of these factors in the development of a disturbed redox status in the kidney cortex.


A review of forty-five years study of Hiroshima and Nagasaki atomic bomb survivors. Tumor registries and cancer incidence studies.
A tumor registry is essential in the systematic collection and analysis of tumor data in a defined population. Population-based tumor registries have been in operation for more than 30 years in Hiroshima and Nagasaki. While the tumor registry data have previously been used for many site-specific cancer studies in atomic bomb survivors, overall analysis of tumor registry based cancer incidence data among the atomic bomb survivors has not been undertaken for some time. However, a recent improvement in the tumor registries has made it possible to analyze most recent cancer incidence data among the Life Span Study population. Preliminary analysis under way indicates a potential power of the data. There are a few methodological problems associated with the use of the incidence data, especially relating to consistency of data and migration. Several measures taken to resolve such problems are discussed.


Survivor But Not Fully Recovered: The Lived Experience After 1 Year of Surviving Sepsis.
Hospital mortality rates related to sepsis have decreased over the last decade, increasing the number of survivors of sepsis who may experience long-term consequences. Yet, little is known about how they perceive their illness experience and its aftermath. This study explores the experiences of sepsis survivors after 1 year of their index intensive care unit (ICU) stay. This phenomenological study was guided by Merleau-Ponty's philosophy. Participants were adult patients admitted directly to the medical ICU with sepsis who had an ICU stay of at least 48 hours and had been discharged from their index ICU stay for at least 1 year. Participants were invited by letter to participate in face-to-face or telephone interviews and a brief survey. Interviews were audio recorded, transcribed verbatim, and verified. An interpretive group read transcripts aloud to increase rigor of identifying meaning units, existential grounds, and thematic structure. Eight participants were recruited over 20 months. Participants were primarily white (87.5%) females (75%) with an age range of 37 to 74 years who were interviewed between 1 and 2 years following an index sepsis ICU stay. Five major themes emerged from the transcripts: (1) how they survived, (2) blurring of time versus counting time by events, (3) helpful help versus unhelpful help, (4) powerless versus striving for control, and (5) survivor but not fully recovered. Findings reflected a long trajectory of chronic illness in which the index episode of sepsis was not necessarily what stood out as figural to participants. The meaning of participant experiences must be considered in the context of a health care system that offers little to no systematic follow-up care after sepsis and little prevention. Participants expressed a need for advocacy and follow-up support.


HIV/AIDS in the small cities: a Brazilian epidemiology study.
Analyzing social, behavior and morbidity data of serum-positive individuals to the test anti-HIV in a Brazilian town city, they were detected: diffusion of the virus of the human immunedeficiency by via heterosexual, in epidemic profile similar to the HIV/AIDS appearance time in the world economically underdeveloped; change of sexual behavior from people when they notice they were involved with HIV; importance of the access to the attendance in the reduction of the morbidity of infected people with HIV and for the transmissibility of this disease in the community. The affected population was between 20 and 49 years and lower socioeconomic level, where the illicit use of drugs was not a risk event, the use of alcoholic drink was an important prominence. The morbidity owed HIV was from low seriousness to moderate one and the answer to the treatment was excellent, with just few collateral effects. Pregnant women had access to the treatment when they were still in good health conditions, but with levels of significant viral load for the vertical HIV transmission.


Renal Tubular CD24 Upregulation Aggravates Folic Acid Induced Acute Kidney Injury: A Possible Role for T Regulatory Cells Inhibition in Mice.
Acute kidney injury (AKI) is characterized by cell death and inflammation. CD24 is a protein induced during tissue damage and is not expressed in mature renal tissue. We explored the role of CD24 in the pathogenesis of folic acid-induced AKI (FA-AKI) in mice. A single Intraperitoneal (IP) injection of folic acid induced AKI in WT and CD24-/- mice. Renal function tests, histological analysis, immunohistochemistry, Western blot analysis, and ELISA were performed to assess the severity of renal damage and the intensity of the inflammatory response. FA-AKI induced CD24 in the distal tubular epithelial cells. Compared to WT mice, FA-AKI CD24-/- mice exhibited an attenuated reduction in renal function and histological injury, lower serum IL-10 and interferon γ, and decreased expression of renal TNFα. In contrast, renal and systemic IL-33 upregulation were augmented. CD24-/- FA-AKI animals exhibited increased splenic margination and renal infiltration of regulatory T cells (Tregs). At day 7, FA-AKI CD24-/- mice exhibited increased expression of tubular pro-apoptotic and decreased anti-apoptotic proteins compared to WT animals. Anti-CD24 antibody administration to FA-AKI mice attenuated the decrease in renal function as well as the histological injury. Renal biopsies from patients with ATN stained strongly for CD24 in the distal tubules. In conclusion, during AKI, upregulation of CD24 promotes renal inflammation through inhibition of Treg infiltration and diversion of cell death towards necrosis rather than apoptosis. Neutralization of CD24 may prove a target for future therapies in AKI.


Reward alters the perception of time.
Recent findings indicate that monetary rewards have a powerful effect on cognitive performance. In order to maximize overall gain, the prospect of earning reward biases visual attention to specific locations or stimulus features improving perceptual sensitivity and processing. The question we addressed in this study is whether the prospect of reward also affects the subjective perception of time. Here, participants performed a prospective timing task using temporal oddballs. The results show that temporal oddballs, displayed for varying durations, presented in a sequence of standard stimuli were perceived to last longer when they signaled a relatively high reward compared to when they signaled no or low reward. When instead of the oddball the standards signaled reward, the perception of the temporal oddball remained unaffected. We argue that by signaling reward, a stimulus becomes subjectively more salient thereby modulating its attentional deployment and distorting how it is perceived in time.


New drugs in psychiatry: focus on new pharmacological targets.
The approval of psychotropic drugs with novel mechanisms of action has been rare in recent years. To address this issue, further analysis of the pathophysiology of neuropsychiatric disorders is essential for identifying new pharmacological targets for psychotropic medications. In this report, we detail drug candidates being examined as treatments for psychiatric disorders. Particular emphasis is placed on agents with novel mechanisms of action that are being tested as therapies for depression, schizophrenia, or Alzheimer's disease. All of the compounds considered were recently approved for human use or are in advanced clinical trials. Drugs included here are new antipsychotic medications endowed with a preferential affinity at dopamine D3 receptor (cariprazine) or at glutamatergic or cannabinoid receptors, as well as vortioxetine, a drug approved for managing the cognitive deficits associated with major depression. New mechanistic approaches for the treatment of depression include intravenous ketamine or esketamine or intranasal esketamine. As for Alzheimer's disease, the possible value of passive immunotherapy with agents such as aducanumab is considered to be a potential disease-modifying approach that could slow or halt the progressive decline associated with this devastating disorder.


Recombinant Human Serum Albumin Containing 3 Copies of Domain I, Has Significant in Vitro Antioxidative Capacity Compared to the Wild-Type.
Human serum albumin (HSA), the most abundant protein in serum, functions as carrier of drugs and contributes to maintaining serum colloid osmotic pressure. We report herein on the preparation of a genetic recombinant HSA, in which domains II and III were changed to domain I (triple domain I; TDI). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results indicated that the purity of the TDI was equivalent to that of the wild type (WT). Both far- and near-UV circular dichroism (CD) spectra of the TDI showed that its structural characteristics were similar to the WT. Ligand binding capacity was examined by an ultrafiltration method using 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF) and ketoprofen as markers for site I and site II, respectively. The binding capacity of TDI for both ligands was lower than that for the wild type. TDI significantly suppressed the oxidation of dihydrorhodamine 123 (DRD) by H2O2 compared to the WT. Our current results suggest that TDI has great potential for further development as HSA a product having antioxidative functions.


Identification of a mammalian target of kappaM-conotoxin RIIIK.
Despite the great variability of the conus peptides characterized until now only relatively few have been identified that interact with K+ channels. kappaM-conotoxin RIIIK (kappaM-RIIIK) is a 24 amino acid peptide from Conus radiatus, which is structurally similar to micro-conotoxin GIIIA, a peptide known to block specifically skeletal muscle Na+ channels. Recently, it has been shown that kappaM-RIIIK does not interact with Na) channels, but inhibits Shaker potassium channels expressed in Xenopus oocytes. It was demonstrated that kappaM-RIIIK binds to the pore region of Shaker channels and a teleost homologue of the Shaker channel TSha1 was identified as a high affinity target of the toxin. In contrast the mammalian Shaker-homologues Kv1.1, Kv1.3, Kv1.4 are not affected by the toxin. In this study the activity of kappaM-RIIIK on other mammalian Kv1 K+ channels expressed in Xenopus oocytes was investigated. We demonstrate that kappaM-conotoxin RIIIK up to 5 microM exhibits no significant effect on Kv1.5 and Kv1.6 mediated currents, but the human Kv1.2 K+ channel is blocked by this peptide. The binding of kappaM-RIIIK to Kv1.2 channels is state dependent with an IC50 for the closed state of about 200 nM and for the open state of about 400 nM at a test potential of 0 mV. kappaM-conotoxin RIIIK is the first conotoxin described to block human Kv1.2 potassium channels.


Overexpression of amyloid beta precursor protein enhances expression and secretion of ST6Gal1 in C2C12 myogenic cell line.
The Abeta (amyloid-beta) peptide is derived from the sequential cleavage of AbetaPP (amyloid-beta precursor protein) by two enzymes, the β- and γ-secretases. The major β-secretase, identified as the novel transmembrane aspartic protease BACE1 (beta site APP-cleaving enzyme 1), mediates the primary amyloidogenic cleavage of AbetaPP and initiates the production of Abeta. It has been implicated in the proteolytic processing of another substrate, namely ST6Gal1 (β galactoside α2,6-sialyltransferase 1), which is the major α2,6-sialyltransferase responsible for the broad synthesis of glycoproteins and glycolipids. The present study investigated the effect of overexpression of AbetaPP on expression and secretion of ST6Gal1 in skeletal muscle cells by inducing overexpression of wild-type full-length 751-AbetaPP in the mouse myogenic cell line C2C12. Expression and secretion of the ST6Gal1 enzyme were analysed by Western blot and/or immunofluorescence staining. The results of our study demonstrated that AbetaPP overexpression in C2C12 cells increased the expression and the secretion of ST6Gal1 enzyme in vitro.


Daily school peer victimization experiences among Mexican-American adolescents: associations with psychosocial, physical and school adjustment.
School bullying incidents, particularly experiences with victimization, are a significant social and health concern among adolescents. The current study extended past research by examining the daily peer victimization experiences of Mexican-American adolescents and examining how chronic (mean-level) and episodic (daily-level) victimization incidents at school are associated with psychosocial, physical and school adjustment. Across a two-week span, 428 ninth and tenth grade Mexican-American students (51 % female) completed brief checklists every night before going to bed. Hierarchical linear model analyses revealed that, at the individual level, Mexican-American adolescents' who reported more chronic peer victimization incidents across the two-weeks also reported heightened distress and academic problems. After accounting for adolescent's mean levels of peer victimization, daily victimization incidents were associated with more school adjustment problems (i.e., academic problems, perceived role fulfillment as a good student). Additionally, support was found for the mediation model in which distress accounts for the mean-level association between peer victimization and academic problems. The results from the current study revealed that everyday peer victimization experiences among Mexican-American high school students have negative implications for adolescents' adjustment, across multiple domains.


Clinical pharmacology of calcium antagonists.
Verapamil, nifedipine, and diltiazem are effective in the treatment of stable effort angina and angiospastic (variant) angina. In addition, there is evidence that the agents are beneficial in patients diagnosed as having unstable angina. The efficacy of calcium antagonists for the treatment of effort angina appears to be augmented by combining them with beta-adrenergic blockers. Intravenous verapamil, but not nifedipine, is very effective in terminating paroxysmal supraventricular tachycardias caused by re-entrant mechanisms with or without involvement of accessory pathways (WPW syndrome). Verapamil is also effective in slowing the ventricular response to atrial fibrillation or flutter. The use of verapamil for the treatment of ectopic atrial or ventricular dysrhythmias is less well established and will require further evaluation.


PD-1 regulates T cell proliferation in a tissue and subset-specific manner during normal mouse pregnancy.
The regulation of T cell homeostasis during pregnancy has important implications for maternal tolerance and immunity. Evidence suggests that Programmed Death-1 (PD-1) participates in regulation of T cell homeostasis and peripheral tolerance. To examine the contribution of PD-1 signaling on T cell homeostasis during normal mouse pregnancy, we examined T cell number or proportion, PD-1 expression, proliferation, and apoptosis by flow cytometry, BrdU incorporation, and TUNEL assay in pregnant mice given anti-PD-1 blocking antibody or control on days 10, 12, and 14 of gestation. We observed tissue, treatment, and T cell-specific differences in PD-1 expression. Both pregnancy and PD-1 blockade increased T cell proliferation in the spleen, yet this effect was limited to CD4 T cells in the uterine- draining nodes. In the uterus, PD-1 blockade markedly altered the composition of the T cell pool. These studies support the idea that pregnancy is a state of dynamic T cell homeostasis and suggest that this state is partially supported by PD-1 signaling.


More epigenetic hits than meets the eye: microRNAs and genes associated with the tumorigenesis of retinoblastoma.
Retinoblastoma (RB), a childhood neoplasia of the retinoblasts, can occur unilaterally or bilaterally, with one or multiple foci per eye. RB is associated with somatic loss of function of both alleles of the tumor suppressor gene RB1. Hereditary forms emerge due to germline loss of function mutations in RB1 alleles. RB has long been the prototypic "model" cancer ever since Knudson's "two-hit" hypothesis. However, a simple two-hit model for RB is challenged by an increasing number of studies documenting additional hits that contribute to RB development. Here we review the genetics and epigenetics of RB with a focus on the role of small non-coding RNAs (microRNAs) and on novel findings indicating the relevance of DNA methylation in the development and prognosis of this neoplasia. Studies point to an elaborated landscape of genetic and epigenetic complexity, in which a number of events and pahtways play crucial roles in the origin and prognosis of RB. These include roles for microRNAs, inprinted loci, and parent-of-origin contributions to RB1 regulation and RB progression. This complexity is also manifested in the structure of the RB1 locus itself: it includes numerous repetitive DNA segments and retrotransposon insertion elements, some of which are actively transcribed from the RB1 locus. Altogether, we conclude that RB1 loss of function represents the tip of an iceberg of events that determine RB development, progression, severity, and disease risk. Comprehensive assessment of personalized RB risk will require genetic and epigenetic evaluations beyond RB1 protein coding sequences.


The mechanism of flecainide action in CPVT does not involve a direct effect on RyR2.
Flecainide, a class 1c antiarrhythmic, has emerged as an effective therapy in preventing arrhythmias in patients with catecholaminergic polymorphic ventricular tachycardia (CPVT) refractory to β-adrenergic receptor blockade. It has been proposed that the clinical efficacy of flecainide in CPVT is because of the combined actions of direct blockade of ryanodine receptors (RyR2) and Na(+) channel inhibition. However, there is presently no direct evidence to support the notion that flecainide blocks RyR2 Ca(2+) flux in the physiologically relevant (luminal-to-cytoplasmic) direction. The mechanism of flecainide action remains controversial. To examine, in detail, the effect of flecainide on the human RyR2 channel and to establish whether the direct blockade of physiologically relevant RyR2 ion flow by the drug contributes to its therapeutic efficacy in the clinical management of CPVT. Using single-channel analysis, we show that, even at supraphysiological concentrations, flecainide did not inhibit the physiologically relevant, luminal-to-cytosolic flux of cations through the channel. Moreover, flecainide did not alter RyR2 channel gating and had negligible effect on the mechanisms responsible for the sarcoplasmic reticulum charge-compensating counter current. Using permeabilized cardiac myocytes to eliminate any contribution of plasmalemmal Na(+) channels to the observed actions of the drug at the cellular level, flecainide did not inhibit RyR2-dependent sarcoplasmic reticulum Ca(2+) release. The principal action of flecainide in CPVT is not via a direct interaction with RyR2. Our data support a model of flecainide action in which Na(+)-dependent modulation of intracellular Ca(2+) handling attenuates RyR2 dysfunction in CPVT.


Rituximab monotherapy for remission induction of proliferative lupus nephritis flares: description of 3 cases.
The efficacy and safety of rituximab when not associated with other immunosuppressive therapy in induction of remission of proliferative lupus nephritis (PLN) has not until now been proven. We report on 3 patients with PLN (class IV), 1 with a nephritic flare and 2 with a proteinuric flare (all with nephrotic syndrome (NS)) treated with 3 intravenous methylprednisolone pulses for 3 consecutive days and with rituximab at day 3 and day 18 associated with oral prednisone. At the beginning of the fourth month of therapy, mycophenolate mofetil was combined with prednisone. Three months after the beginning of therapy, renal function improved in the patient with the nephritic flare, and proteinuria fell to within the non-nephrotic range in all 3 patients. At the end of a follow-up of 24 months, the patient with the nephritic flare had normal renal function (serum creatinine from 1.7 to 1 mg/dL) and mild proteinuria (from 6 to 0.7 g/24 hours). The second patient was in complete remission (proteinuria from 5 to 0.127 g/24 hours) 27 months after the beginning of therapy. In the last patient, followed for 10 months, mild proteinuria persisted (from 6.6 to 0.7 g/24 hours). The therapy was well tolerated by all patients. No adverse effects occurred during the follow-up. Although our results must be confirmed by larger prospective studies, rituximab associated with methylprednisolone pulses without any other concomitant immunosuppressive drug seems to be effective and safe for induction therapy of severe flares of PLN.


Sonographic screening for urinary tract abnormalities in patients with Schistosoma haematobium infection: pitfalls in examining pregnant women.
In areas where Schistosoma haematobium is endemic, urinary schistosomiasis and pregnancy are frequently concomitant; however, both these conditions may produce similar urinary tract changes in ultrasound scans and hence their differential diagnosis may be difficult. In patients with urinary schistosomiasis, focal and/or diffuse urinary bladder wall changes are frequently detected ultrasonically. Dilatation of one or both ureters and progressive hydronephrosis may be observed in more severe cases. Satisfactory ultrasound examination of the urinary bladders of pregnant women is generally not feasible because mechanical compression by the fetus or transitory lower urinary tract infection hampers adequate filling of the bladder. Pregnancy itself is frequently associated with dilatation of one or both ureters and/or hydronephrosis; this is due to hormonal factors, infection, or compression of one or both ureters by the enlarged uterus and growing fetus. Hence, when sonography of the urinary bladder is not feasible such pregnancy-associated changes are virtually indistinguishable from those caused by S. haematobium, and may be incorrectly attributed to the latter. Pregnant women, therefore, should be excluded from ultrasonic surveys of urinary schistosomiasis. In contrast, ultrasound scans of adolescents and of women with positive parasitological findings and/or pathological alterations in the urinary tract should include examination of the uterus in order to assess whether the woman is pregnant; thereby, misinterpretation of sonographic findings can be avoided. Pregnant women with significant hydronephrosis must be closely followed up by an obstetrician since this condition may indicate a complication of the pregnancy; in some cases only a postpartum examination will permit definitive diagnosis.


Arterial baroreflex regulation of regional vascular conductance at rest and during exercise.
We tested the hypothesis that dynamic exercise resets the operating point and attenuates the spontaneous gain of the arterial baroreflex regulation of mesenteric and hindlimb vascular conductance in hypertensive rats. Eleven adult male spontaneously hypertensive rats were chronically instrumented with left carotid arterial catheters and Doppler ultrasonic flow probes around the superior mesenteric and left common iliac arteries. After the rats recovered, arterial baroreflex function was examined by recording reflex changes in conductance in response to spontaneous changes in mean arterial pressure before exercise and during steady-state treadmill running at 6 and 18 m/min. Dynamic exercise reduced the spontaneous baroreflex gain of mesenteric conductance (by 51 and 36%) and maximum mesenteric conductance (by 24 and 32%) at 6 and 18 m/min, respectively. In sharp contrast, dynamic exercise increased the spontaneous maximum iliac conductance (by 32 and 47%) without changing the spontaneous gain. Sinoaortic denervation eliminated the relationship between mean arterial pressure and conductance by reducing the mesenteric (92%) and iliac (68%) vascular conductance gain. These results demonstrate that dynamic exercise has differential effects on the regulation of mesenteric and iliac vascular conductance in hypertensive rats.


Bioinformatics analysis of bacterial annexins--putative ancestral relatives of eukaryotic annexins.
Annexins are Ca(2+)-binding, membrane-interacting proteins, widespread among eukaryotes, consisting usually of four structurally similar repeated domains. It is accepted that vertebrate annexins derive from a double genome duplication event. It has been postulated that a single domain annexin, if found, might represent a molecule related to the hypothetical ancestral annexin. The recent discovery of a single-domain annexin in a bacterium, Cytophaga hutchinsonii, apparently confirmed this hypothesis. Here, we present a more complex picture. Using remote sequence similarity detection tools, a survey of bacterial genomes was performed in search of annexin-like proteins. In total, we identified about thirty annexin homologues, including single-domain and multi-domain annexins, in seventeen bacterial species. The thorough search yielded, besides the known annexin homologue from C. hutchinsonii, homologues from the Bacteroidetes/Chlorobi phylum, from Gemmatimonadetes, from beta- and delta-Proteobacteria, and from Actinobacteria. The sequences of bacterial annexins exhibited remote but statistically significant similarity to sequence profiles built of the eukaryotic ones. Some bacterial annexins are equipped with additional, different domains, for example those characteristic for toxins. The variation in bacterial annexin sequences, much wider than that observed in eukaryotes, and different domain architectures suggest that annexins found in bacteria may actually descend from an ancestral bacterial annexin, from which eukaryotic annexins also originate. The hypothesis of an ancient origin of bacterial annexins has to be reconciled with the fact that remarkably few bacterial strains possess annexin genes compared to the thousands of known bacterial genomes and with the patchy, anomalous phylogenetic distribution of bacterial annexins. Thus, a massive annexin gene loss in several bacterial lineages or very divergent evolution would appear a likely explanation. Alternative evolutionary scenarios, involving horizontal gene transfer between bacteria and protozoan eukaryotes, in either direction, appear much less likely. Altogether, current evidence does not allow unequivocal judgement as to the origin of bacterial annexins.


Unidirectional complexation of pillar[4]arene[1]benzoquinoneoxime with alkyl alcohols.
Despite progress in the unidirectional complexation of cyclodextrins and calixarenes with nonsymmetric guests, unidirectional complexation using pillararenes as hosts remains scarcely explored. In this report, we describe the formation of unidirectional [2]pseudorotaxane-like complexes which were realized using n-alkyl alcohol guests and pillar[4]arene[1]benzoquinoneoxime made from pillar[4]arene[1]quinone in a selective monofunctionalizing manner.


Involvement of 1,2-diacylglycerol in improvement of heart function by etomoxir in diabetic rats.
Abnormal lipid metabolism has been proposed to be involved in the pathogenesis of diabetic cardiomyopathy. In this study, we measured myocardial lipid levels, including 1,2-diacylglycerol (1,2-DAG) and ceramide (CM), and myocardial function in diabetic rats. We also evaluated the effects of etomoxir (ETM), a carnitine palmitoyl transferase I inhibitor, on diabetic rat hearts from the viewpoints of alterations in lipid second messengers and myocardial function. Rats were injected with streptozotocin (60 mg/kg) to induce diabetes and were treated 5 weeks later with ETM (18 mg/kg) for 8 days. In diabetic rats, heart rate, systolic blood pressure, and fractional shortening were significantly reduced compared with those in controls. Treatment of diabetic rats with ETM ameliorated myocardial dysfunction other than heart rate. Myocardial 1,2-DAG levels in diabetic rats were significantly elevated compared with those in controls, while myocardial CM levels were not. ETM treatment caused an additional increase in myocardial 1,2-DAG levels in diabetic rats, but the CM levels did not change. There was a marked difference in fatty acid pattern of 1,2-DAG between diabetic and ETM-treated diabetic rat hearts. The fatty acids 18:1 and 18:2 were significantly increased and the fatty acids 16:0, 18:0, 20:4, and 22:6 were significantly reduced in ETM-treated diabetic rat hearts. These data suggest 1,2-DAG is involved in ameliorating myocardial dysfunction in diabetic rats and that its source is different between diabetic and ETM-treated diabetic rats. CM is unlikely to be involved in the pathogenesis of diabetic cardiomyopathy or the improvement of cardiac contractility in diabetic rats by ETM.


A molecular imprinted polymer based sensor for measuring phosphate in wastewater samples.
Phosphate detection in water samples is still completed using colorimetric standard methods, which have a number of disadvantages, to such as being time consuming, requiring filtration, a number of different reagents, frequent calibration and proper disposal of waste chemicals generated. Hence, a simple cost effective analytical method and instrumentation is highly desirable to aid the optimisation of treatment processes and assist the water industry in their efforts to comply with stringent regulations such as the EU's Water Framework Directive. A sensor based on molecular imprinted polymer (MIP) and a conductance transducer was developed for direct and label-free detection of phosphate in water. The sensor was able to measure the presence of phosphate in wastewater samples with good reproducibility, a linear range of 0.66-8 mg P L(-1) and a lower detection limit of 0.16 mg P L(-1). The sensor was further tested to measure phosphate concentrations in unfiltered field samples such as domestic wastewater treatment influent and river water and demonstrated a close correlation with reference measurements. The phosphate MIP sensor offers a way forward as either a handheld sensor for use in the field, or as a potential solution for remote, continuous monitoring of phosphate.


Precision medicine in asthma: linking phenotypes to targeted treatments.
Asthma is a heterogeneous disease consisting of different phenotypes that are driven by different mechanistic pathways. The purpose of this review is to emphasize the important role of precision medicine in asthma management. Despite asthma heterogeneity, the approach to management has been on the basis of disease severity, with the most severe patients reserved for the maximum treatments with corticosteroids and bronchodilators. At the severe end, the recent availability of biologic therapies in the form of anti-IgE (omalizumab) and anti-IL5 therapies (mepolizumab and reslizumab) has driven the adaptation of precision medicine. These therapies are reserved for severe asthma with defined either allergic or eosinophilic background, respectively. Unbiased definition of phenotypes or endotypes (which are phenotypes defined by mechanisms) is an important step towards the use of precision medicine in asthma. Although T2-high asthma has been defined with targets becoming available for treating allergic or eosinophilic asthma, the definition of non-T2 phenotypes remains a priority. Precision medicine is also dependent on the definition of biomarkers that can help differentiate between these phenotypes and pinpoint patients suitable for specific-targeted therapies. Thus, precision medicine links phenotypes (endotypes) to targeted treatments for better outcomes.


Interaction of lubricin with type II collagen surfaces: adsorption, friction, and normal forces.
One of the major constituents of the synovial fluid that is thought to be responsible for chondroprotection and boundary lubrication is the glycoprotein lubricin (PRG4); however, the molecular mechanisms by which lubricin carries out its critical functions still remain largely unknown. We hypothesized that the interaction of lubricin with type II collagen, the main component of the cartilage extracellular matrix, results in enhanced tribological and wear properties. In this study, we examined: (i) the molecular details by which lubricin interacts with type II collagen and how binding is related to boundary lubrication and adhesive interactions; and (ii) whether collagen structure can affect lubricin adsorption and its chondroprotective properties. We found that lubricin adsorbs strongly onto denatured, amorphous, and fibrillar collagen surfaces. Furthermore, we found large repulsive interactions between the collagen surfaces in presence of lubricin, which increased with increasing lubricin concentration. Lubricin attenuated the large friction and also the long-range adhesion between fibrillar collagen surfaces. Interestingly, lubricin adsorbed onto and mediated the frictional response between the denatured and native amorphous collagen surfaces equally and showed no preference on the supramolecular architecture of collagen. However, the coefficient of friction was lowest on fibrillar collagen in the presence of lubricin. We speculate that an important role of lubricin in mediating interactions at the cartilage surface is to attach to the cartilage surface and provide a protective coating that maintains the contacting surfaces in a sterically repulsive state.


Targeting PES1 for restoring the ERα/ERβ ratio in breast cancer.
Alteration of the ERα/ERβ balance is a critical step in breast cancer development and progression, and selective restoration of the activity of estrogen receptors has been proposed as one of the major therapeutic approaches for breast cancer. In this issue of JCI, Cheng et al. show that, by differentially modulating the stability of ERα and ERβ, PES1 increases the ERα/ERβ ratio and triggers breast tumor growth. These findings highlight PES1 as a potential target for the treatment of breast cancer.


Protein ligands to HuR modulate its interaction with target mRNAs in vivo.
AU-rich elements (AREs) present in the 3' untranslated regions of many protooncogene, cytokine, and lymphokine messages target them for rapid degradation. HuR, a ubiquitously expressed member of the ELAV (embryonic lethal abnormal vision) family of RNA binding proteins, selectively binds AREs and stabilizes ARE-containing mRNAs in transiently transfected cells. Here, we identify four mammalian proteins that bind regions of HuR known to be essential for its ability to shuttle between the nucleus and the cytoplasm and to stabilize mRNA: SETalpha, SETbeta, pp32, and acidic protein rich in leucine (APRIL). Three have been reported to be protein phosphatase 2A inhibitors. All four ligands contain long, acidic COOH-terminal tails, while pp32 and APRIL share a second motif: rev-like leucine-rich repeats in their NH(2)-terminal regions. We show that pp32 and APRIL are nucleocytoplasmic shuttling proteins that interact with the nuclear export factor CRM1 (chromosomal region maintenance protein 1). The inhibition of CRM1 by leptomycin B leads to the nuclear retention of pp32 and APRIL, their increased association with HuR, and an increase in HuR's association with nuclear poly(A)+ RNA. Furthermore, transcripts from the ARE-containing c-fos gene are selectively retained in the nucleus, while the cytoplasmic distribution of total poly(A)+ RNA is not altered. These data provide evidence that interaction of its ligands with HuR modulate HuR's ability to bind its target mRNAs in vivo and suggest that CRM1 is instrumental in the export of at least some cellular mRNAs under certain conditions. We discuss the possible role of these ligands upstream of HuR in pathways that govern the stability of ARE-containing mRNAs.


First evidence for a covalent linkage between enterobacterial common antigen and lipopolysaccharide in Shigella sonnei phase II ECALPS.
Enterobacterial common antigen (ECA) is expressed by Gram-negative bacteria belonging to Enterobacteriaceae, including emerging drug-resistant pathogens such as Escherichia coli, Klebsiella pneumoniae, and Proteus spp. Recent studies have indicated the importance of ECA for cell envelope integrity, flagellum expression, and resistance of enteric bacteria to acetic acid and bile salts. ECA, a heteropolysaccharide built from the trisaccharide repeating unit, →3)-α-D-Fucp4NAc-(1→4)-β-D-ManpNAcA-(1→4)-α-D-GlcpNAc-(1→, occurs as a cyclic form (ECA(CYC)), a phosphatidylglycerol (PG)-linked form (ECA(PG)), and an endotoxin/lipopolysaccharide (LPS)-associated form (ECA(LPS)). Since the discovery of ECA in 1962, the structures of ECA(PG) and ECA(CYC) have been completely elucidated. However, no direct evidence has been presented to support a covalent linkage between ECA and LPS; only serological indications of co-association have been reported. This is paradoxical, given that ECA was first identified based on the capacity of immunogenic ECA(LPS) to elicit antibodies cross-reactive with enterobacteria. Using a simple isolation protocol supported by serological tracking of ECA epitopes and NMR spectroscopy and mass spectrometry, we have succeeded in the first detection, isolation, and complete structural analysis of poly- and oligosaccharides of Shigella sonnei phase II ECA(LPS). ECA(LPS) consists of the core oligosaccharide substituted with one to four repeating units of ECA at the position occupied by the O-antigen in the case of smooth S. sonnei phase I. These data represent the first structural evidence for the existence of ECA(LPS) in the half-century since it was first discovered and provide insights that could prove helpful in further structural analyses and screening of ECA(LPS) among Enterobacteriaceae species.


Human 6-sulfo LacNAc (slan) dendritic cells are a major population of dermal dendritic cells in steady state and inflammation.
The immune system of the skin has a network of resident dendritic cells (DCs) consisting of epidermal Langerhans cells and various subsets of dermal DCs. We recently reported on a new population of dermal DCs, called slan (6-sulfoLacNAc+) DCs, which have a potent capacity to stimulate Th17/Th1 T-cell responses. To understand the characteristics of slanDCs as a new population of dermal DCs in the context of other DC populations in healthy and psoriatic skin. We immunofluorescently stained skin samples from healthy controls and from patients with psoriasis. Staining healthy skin for DCs showed that slanDCs (CD1a- CD1c- CD11c- CD14- CD163-) were present at a similar frequency to that of CD1c+ CD11c+ CD1a+ CD14- CD163- dermal DCs, which have previously been regarded as the major population of resident DCs. In psoriatic skin, the frequency of slanDCs and CD1c+ DCs was doubled, and the slanDCs expressed CD11c. In-depth analysis of DCs in psoriatic skin by four-colour immunofluorescence analysis showed that the pool of CD11c+ cells could be further subdivided into CD11c+ CD14+ CD163- DCs and CD11c+ CD163+ CD14+ macrophages. SlanDCs, initially described as large population of proinflammatory DCs in blood, are a novel and major part of the resident dermal myeloid DC system in both healthy and inflamed skin.


Risk of violence by psychiatric patients: beyond the "actuarial versus clinical" assessment debate.
Recently adopted statistical approaches improve researchers' ability to describe what is, and what is not, possible in the prediction of violence by psychiatric patients. At the base rates of violence routinely encountered in outpatient settings, current assessment methods would require hospital admission of large numbers of patients who are potential offenders in order to prevent the actual offending of a few. Suggestions that substantially greater accuracy is possible for short-term predictions, for particular symptom clusters, and for particular offenses have yet to be tested and confirmed. Further research may improve this state of affairs, for instance, by concentrating on particular patient groups. There are reasons to suspect that any improvement will be limited. Clinical practice, however, is likely to continue to require the assessment of a patient's potential for acting violently. Future research should aid such assessments by clarifying the mechanisms by which risk factors correlate with violence and by establishing the clinical usefulness of actuarial scales.


In vitro and in vivo antimalarial evaluations of myrtle extract, a plant traditionally used for treatment of parasitic disorders.
Based on the collected ethnobotanical data from the Traditional Medicine and Materia Medica Research Center (TMRC), Iran, Myrtus communis L. (myrtle) was selected for the assessment of in vitro and in vivo antimalarial and cytotoxic activities. Methanolic extract of myrtle was prepared from the aerial parts and assessed for antiplasmodial activity, using the parasite lactate dehydrogenase (pLDH) assay against chloroquine-resistant (K1) and chloroquine-sensitive (3D7) strains of Plasmodium falciparum. The 4-day suppressive test was employed to determine the parasitemia suppression of the myrtle extract against P. berghei in vivo. The IC50 values of myrtle extract were 35.44 µg/ml against K1 and 0.87 µg/ml against 3D7. Myrtle extract showed a significant suppression of parasitaemia (84.8 ± 1.1% at 10 mg/kg/day) in mice infected with P. berghei after 4 days of treatment. Cytotoxic activity was carried out against mammalian cell lines using methyl thiazol tetrazolium (MTT) assay. No cytotoxic effect on mammalian cell lines up to 100 µg/mL was shown. The results support the traditional use of myrtle in malaria. Phytochemical investigation and understanding the mechanism of action would be in our upcoming project.


Duration-dependent changes in calcium responsiveness in the alloxan-diabetic rat intestine.
Disturbances of the gastrointestinal tract that are common in diabetes mellitus seem to be related to intestinal motility. In experimental models of diabetes, decreased calcium sensitivity has been demonstrated in various smooth muscles including those in the gastrointestinal tract. The main purpose of the present study was to examine further the calcium sensitivity in diabetic rat intestine and to understand if changes in the calcium sensitivity occur at an earlier stage of the disease. For this purpose, the effects of potassium and calcium were evaluated on nondepolarized and depolarized duodenum from rats with alloxan diabetes for 1 and 8 weeks and their age-matched controls. To evaluate the calcium sensitivity in rat duodenum, apparent affinity constants (pD2 values) and intrinsic activities (alpha E values) were calculated for every experimental conditions examined in this study. Both values (pD2 and alpha E) for the effects of potassium and calcium on the nondepolarized and depolarized duodenum, respectively, were not changed in 1-week diabetic rats. In contrast, intrinsic activities for the effects of potassium and calcium were found to be significantly decreased (p < 0.001) in the nondepolarized and depolarized duodenum from rats with alloxan diabetes for 8 weeks, whereas apparent affinity constants were not altered in this case. Taking into consideration all these experimental findings, the decreased calcium sensitivity in gastrointestinal tract seems to be closely related to decreased calmodulin levels and may occur at a later stage of diabetes as a linkage to long-term gastrointestinal complications.


Southern Ocean in-situ temperature trends over 25 years emerge from interannual variability.
Despite playing a major role in global ocean heat storage, the Southern Ocean remains the most sparsely measured region of the global ocean. Here, a unique 25-year temperature time-series of the upper 800 m, repeated several times a year across the Southern Ocean, allows us to document the long-term change within water-masses and how it compares to the interannual variability. Three regions stand out as having strong trends that dominate over interannual variability: warming of the subantarctic waters (0.29 ± 0.09 °C per decade); cooling of the near-surface subpolar waters (-0.07 ± 0.04 °C per decade); and warming of the subsurface subpolar deep waters (0.04 ± 0.01 °C per decade). Although this subsurface warming of subpolar deep waters is small, it is the most robust long-term trend of our section, being in a region with weak interannual variability. This robust warming is associated with a large shoaling of the maximum temperature core in the subpolar deep water (39 ± 09 m per decade), which has been significantly underestimated by a factor of 3 to 10 in past studies. We find temperature changes of comparable magnitude to those reported in Amundsen-Bellingshausen Seas, which calls for a reconsideration of current ocean changes with important consequences for our understanding of future Antarctic ice-sheet mass loss.


Precise segmentation of multimodal images.
We propose new techniques for unsupervised segmentation of multimodal grayscale images such that each region-of-interest relates to a single dominant mode of the empirical marginal probability distribution of grey levels. We follow the most conventional approaches in that initial images and desired maps of regions are described by a joint Markov-Gibbs random field (MGRF) model of independent image signals and interdependent region labels. However, our focus is on more accurate model identification. To better specify region borders, each empirical distribution of image signals is precisely approximated by a linear combination of Gaussians (LCG) with positive and negative components. We modify an expectation-maximization (EM) algorithm to deal with the LCGs and also propose a novel EM-based sequential technique to get a close initial LCG approximation with which the modified EM algorithm should start. The proposed technique identifies individual LCG models in a mixed empirical distribution, including the number of positive and negative Gaussians. Initial segmentation based on the LCG models is then iteratively refined by using the MGRF with analytically estimated potentials. The convergence of the overall segmentation algorithm at each stage is discussed. Experiments show that the developed techniques segment different types of complex multimodal medical images more accurately than other known algorithms.


Pharmacokinetics and Antifungal Activity of Echinocandins in Ascites Fluid of Critically Ill Patients.
The pharmacokinetics and antifungal activity of the echinocandins anidulafungin (AFG), micafungin (MFG), and caspofungin (CAS) were assessed in ascites fluid and plasma of critically ill adults treated for suspected or proven invasive candidiasis. Ascites fluid was obtained from ascites drains or during paracentesis. The antifungal activity of the echinocandins in ascites fluid was assessed by incubation of Candida albicans and Candida glabrata at concentrations of 0.03 to 16.00 μg/ml. In addition, ascites fluid samples obtained from our study patients were inoculated with the same isolates and evaluated for fungal growth. These patient samples had to be spiked with echinocandins to restore the original concentrations because echinocandins had been lost during sterile filtration. In ascites fluid specimens of 29 patients, echinocandin concentrations were below the simultaneous plasma levels. Serial sampling in 20 patients revealed a slower rise and decline of echinocandin concentrations in ascites fluid than in plasma. Proliferation of C. albicans in ascites fluid was slower than in culture medium and growth of C. glabrata was lacking, even in the absence of antifungals. In CAS-spiked ascites fluid samples, fungal CFU counts moderately declined, whereas spiking with AFG or MFG had no relevant effect. In ascites fluid of our study patients, echinocandin concentrations achieved by therapeutic doses did not result in a consistent eradication of C. albicans or C. glabrata. Thus, therapeutic doses of AFG, MFG, or CAS may result in ascites fluid concentrations preventing relevant proliferation of C. albicans and C. glabrata, but do not warrant reliable eradication.


Do Social Protection Programs Foster Short-term and Long-term Migration Adaptation Strategies?
We examine how migration is influenced by temperature and precipitation variability, and the extent to which the receipt of a cash transfer affects the use of migration as an adaptation strategy. Climate data is merged with georeferenced panel data (2010-2014) on individual migration collected from the Zambian Child Grant Program (CGP) sites. We use the person-year dataset to identify the direct and heterogeneous causal effects of the CGP on mobility. Having access to cash transfers doubles the rate of male, short-distance moves during cool periods irrespective of wealth. Receipt of cash transfers (among wealthier households) during extreme heat causes an additional retention of males. Cash transfers positively spur long-distance migration under normal climate conditions in the long term. They also facilitate short-distance responses to climate, but not long-distance responses that might be demanded by future climate change.


An allele-sharing, moment-based estimator of global, population-specific and population-pair FST under a general model of population structure.
Being able to properly quantify genetic differentiation is key to understanding the evolutionary potential of a species. One central parameter in this context is FST, the mean coancestry within populations relative to the mean coancestry between populations. Researchers have been estimating FST globally or between pairs of populations for a long time. More recently, it has been proposed to estimate population-specific FST values, and population-pair mean relative coancestry. Here, we review the several definitions and estimation methods of FST, and stress that they provide values relative to a reference population. We show the good statistical properties of an allele-sharing, method of moments based estimator of FST (global, population-specific and population-pair) under a very general model of population structure. We point to the limitation of existing likelihood and Bayesian estimators when the populations are not independent. Last, we show that recent attempts to estimate absolute, rather than relative, mean coancestry fail to do so.


Cervical radiculopathy: the diagnosis and treatment of pathomechanics in the cervical spine.
Lateral flexion anteroposterior roentgenograms of the cervical spine were obtained of patients suffering from cervical radiculopathy. Specific attention was given to the normal mechanical attitudes of the cervical motion segments at their end range. Deviations from the normal coupling mechanism in the cervical spine were directly related to the level of clinical radiculopathy. Aberrancies in the coupling mechanism of the cervical motion segments were reduced by manipulation, and post manipulative reduction examination revealed improvements in sensation, motor power, and deep tendon reflexes, as well as subjective patient improvement. Normal mechanical attitudes could be restored by manipulative reduction of the cervical motion segments.


The tumor suppressor gene FBXW7 is disrupted by a constitutional t(3;4)(q21;q31) in a patient with renal cell cancer.
FBXW7 (alias CDC4) is a p53-dependent tumor suppressor gene that exhibits mutations or deletions in a variety of human tumors. Mutation or deletion of the FBXW7 gene has been associated with an increase in chromosomal instability and cell cycle progression. In addition, the FBXW7 protein has been found to act as a component of the ubiquitin proteasome system and to degrade several oncogenic proteins that function in cellular growth regulatory pathways. By using a rapid breakpoint cloning procedure in a case of renal cell cancer (RCC), we found that the FBXW7 gene was disrupted by a constitutional t(3;4)(q21;q31). Subsequent analysis of the tumor tissue revealed the presence of several anomalies, including loss of the derivative chromosome 3. Upon screening of a cohort of 29 independent primary RCCs, we identified one novel pathogenic mutation, suggesting that the FBXW7 gene may also play a role in the development of sporadic RCCs. In addition, we screened a cohort of 48 unrelated familial RCC cases with unknown etiology. Except for several known or benign sequence variants such as single nucleotide polymorphisms (SNPs), no additional pathogenic variants were found. Previous mouse models have suggested that the FBXW7 gene may play a role in the predisposition to tumor development. Here we report that disruption of this gene may predispose to the development of human RCC.