Identification of chikungunya virus nsP2 protease inhibitors using structure-base approaches.
The nsP2 protease of chikungunya virus (CHIKV) is one of the essential components of viral replication and it plays a crucial role in the cleavage of polyprotein precursors for the viral replication process. Therefore, it is gaining attention as a potential drug design target against CHIKV. Based on the recently determined crystal structure of the nsP2 protease of CHIKV, this study identified potential inhibitors of the virus using structure-based approaches with a combination of molecular docking, virtual screening and molecular dynamics (MD) simulations. The top hit compounds from database searching, using the NCI Diversity Set II, with targeting at five potential binding sites of the nsP2 protease, were identified by blind dockings and focused dockings. These complexes were then subjected to MD simulations to investigate the stability and flexibility of the complexes and to gain a more detailed insight into the interactions between the compounds and the enzyme. The hydrogen bonds and hydrophobic contacts were characterized for the complexes. Through structural alignment, the catalytic residues Cys1013 and His1083 were identified in the N-terminal region of the nsP2 protease. The absolute binding free energies were estimated by the linear interaction energy approach and compared with the binding affinities predicted with docking. The results provide valuable information for the development of inhibitors for CHIKV.


5-(Prop-2-yn-1-yl)-5H-dibenzo[b,f]azepine: ortho-rhom-bic polymorph.
In the title ortho-rhom-bic polymorph (space group Iba2), C17H13N, the dihedral angle between the benzene rings is 55.99 (10)° and the azepine ring adopts a boat conformation. In the crystal, mol-ecules are linked by C-H⋯π contacts. The previously-reported polymorph [Yousuf et al. (2012 ▶). Acta Cryst. E68, o1101] crystallizes in the monoclinic system (space group P21/c) with two mol-ecules in the asymmetric unit.


An Evaluation of Ultrasound-Guided Adductor Canal Blockade for Postoperative Analgesia After Medial Unicondylar Knee Arthroplasty.
Unicondylar knee arthroplasty (UKA) is a commonly performed procedure with significant expected postoperative pain. Peripheral nerve blocks are 1 analgesic option, but some approaches may decrease quadriceps motor strength and interfere with early ambulation. In this study, we compared the analgesia provided by an adductor canal block (ACB) and a psoas compartment block (PCB) after UKA. We hypothesized that the ACB would provide equivalent analgesia, defined as a difference of <2 points on the pain scale (0-10 numeric rating scale [NRS]), at rest and with movement 6 hours after block placement. One hundred fifty patients undergoing medial UKA were randomly assigned to receive either an ACB or a PCB with 0.25% bupivacaine, 5 μg/mL epinephrine, and 1.67 μg/mL clonidine. All patients received multimodal analgesics, sham blockade at the alternate site, and a posterior capsule injection during surgery. Patients and observers were blinded to treatment groups. The primary end points were NRS pain scores with rest and movement at 6 hours. Secondary end points included quadriceps muscle strength at 6 hours (0-5 [5 being full strength]; Medical Research Council scale) as well as NRS pain scores, opioid consumption, and opioid-related side effects over 24 hours. One hundred forty-seven patients were analyzed. Pain scores were equivalent at 6 hours with rest (ACB 1.0 ± 2 vs PCB 1.1 ± 2.2 [mean NRS ± SD]; 95% confidence interval of mean difference, -0.8 to 0.6; P < 0.0001) and with movement (ACB 1.6 ± 2.6 vs PCB 1.5 ± 2.8; 95% confidence interval of mean difference, -0.8 to 0.9; P < 0.0001). In addition, pain scores at rest and with movement at 12, 18, and 24 hours were equivalent. Quadriceps motor strength was significantly increased in the ACB group (Medical Research Council scale score, 4.0 ± 1.1 vs 2.5 ± 1.3 [mean ± SD]; P < 0.0001). No significant differences were found between groups for time to first analgesic or for cumulative opioid consumption at 6, 12, 18, or 24 hours. Other than an increase in the incidence of pruritus in the ACB group at 6 hours, there were no differences in opioid-related side effects. An ACB provides equivalent analgesia after medial UKA when compared with a PCB. In addition, the ACB caused significantly less motor weakness. An ACB should be considered for postoperative analgesia after medial UKA.


Increasing childhood vaccination coverage of the refugee and migrant population in Greece through the European programme PHILOS, April 2017 to April 2018.
After the 2016 Balkan route border closures, vaccination of refugee children in Greece was mainly performed by non-governmental organisations. Activities varied between camps, resulting in heterogeneity of vaccination coverage (VC). In April 2017, the European programme 'PHILOS - Emergency health response to refugee crisis' took over vaccination coordination. Interventions were planned for the first time for refugee children in the community and unaccompanied minors at safe zones. From April 2017-April 2018, 57,615 vaccinations were performed against measles-mumps-rubella (MMR) (21,031), diphtheria-tetanus-pertussis (7,341), poliomyelitis (7,652), pneumococcal disease (5,938), Haemophilus influenzae type b (7,179) and hepatitis B (8,474). In April 2018, the vaccination status of children at camps (reception and identification centres and community facilities such as hostels/hotels were excluded) was recorded and VC for each disease, stratified by dose, nationality and camp size, was calculated. More than 80% of the children received the first MMR dose, with VC dropping to 45% for the second dose. For all other vaccines, VC was < 50% for the first dose in children aged 0-4 years and < 25% for the second dose. Despite challenges, PHILOS improved planning and monitoring of vaccination activities; however, further efforts towards improving VC in refugee children are needed.


Structure activity relationship of dendrimer microbicides with dual action antiviral activity.
Topical microbicides, used by women to prevent the transmission of HIV and other sexually transmitted infections are urgently required. Dendrimers are highly branched nanoparticles being developed as microbicides. However, the anti-HIV and HSV structure-activity relationship of dendrimers comprising benzyhydryl amide cores and lysine branches, and a comprehensive analysis of their broad-spectrum anti-HIV activity and mechanism of action have not been published. Dendrimers with optimized activity against HIV-1 and HSV-2 were identified with respect to the number of lysine branches (generations) and surface groups. Antiviral activity was determined in cell culture assays. Time-of-addition assays were performed to determine dendrimer mechanism of action. In vivo toxicity and HSV-2 inhibitory activity were evaluated in the mouse HSV-2 susceptibility model. Surface groups imparting the most potent inhibitory activity against HIV-1 and HSV-2 were naphthalene disulfonic acid (DNAA) and 3,5-disulfobenzoic acid exhibiting the greatest anionic charge and hydrophobicity of the seven surface groups tested. Their anti-HIV-1 activity did not appreciably increase beyond a second-generation dendrimer while dendrimers larger than two generations were required for potent anti-HSV-2 activity. Second (SPL7115) and fourth generation (SPL7013) DNAA dendrimers demonstrated broad-spectrum anti-HIV activity. However, SPL7013 was more active against HSV and blocking HIV-1 envelope mediated cell-to-cell fusion. SPL7013 and SPL7115 inhibited viral entry with similar potency against CXCR4-(X4) and CCR5-using (R5) HIV-1 strains. SPL7013 was not toxic and provided at least 12 h protection against HSV-2 in the mouse vagina. Dendrimers can be engineered with optimized potency against HIV and HSV representing a unique platform for the controlled synthesis of chemically defined multivalent agents as viral entry inhibitors. SPL7013 is formulated as VivaGel(R) and is currently in clinical development to provide protection against HIV and HSV. SPL7013 could also be combined with other microbicides.


Sex Specification and Heterogeneity of Primordial Germ Cells in Mice.
In mice, primordial germ cells migrate into the genital ridges by embryonic day 13.5 (E13.5), where they are then subjected to a sex-specific fate with female and male primordial germ cells undergoing mitotic arrest and meiosis, respectively. However, the sex-specific basis of primordial germ cell differentiation is poorly understood. The aim of this study was to investigate the sex-specific features of mouse primordial germ cells. We performed RNA-sequencing (seq) of E13.5 female and male mouse primordial germ cells using next-generation sequencing. We identified 651 and 428 differentially expressed transcripts (>2-fold, P < 0.05) in female and male primordial germ cells, respectively. Of these, many transcription factors were identified. Gene ontology and network analysis revealed differing functions of the identified female- and male-specific genes that were associated with primordial germ cell acquisition of sex-specific properties required for differentiation into germ cells. Furthermore, DNA methylation and ChIP-seq analysis of histone modifications showed that hypomethylated gene promoter regions were bound with H3K4me3 and H3K27me3. Our global transcriptome data showed that in mice, primordial germ cells are decisively assigned to a sex-specific differentiation program by E13.5, which is necessary for the development of vital germ cells.


High-risk prostate cancer: the role of radical prostatectomy for local therapy.
The management of high-risk prostate cancer can pose a unique challenge to the urologic oncologist. High-risk prostate cancer remains a real entity, especially in the inner-city urban population centers with high-risk ethnic groups. Although the role of radical prostatectomy is well defined for localized, low-to-intermediate-risk prostate cancer, its role in high-risk disease is more controversial. This is compounded by a lack of a universally accepted definition for 'high-risk' disease and the stage migration that has occurred in prostate cancer in the PSA era, rendering some historical perspectives less relevant. However, what has been accepted is the role of multimodal therapy in the management of this challenging group of patients. This article offers the reader an up-to-date detailed review of this topic, with specific emphasis on the role of radical prostatectomy in this clinical setting, including surgical considerations and outcomes. The advantages in terms of accurate pathologic staging with radical prostatectomy are presented. The role of robotic radical prostatectomy, which is increasingly utilized in the USA for the surgical treatment of prostate cancer in this clinical scenario, is discussed. In addition, we address the shortcomings of adequate clinical staging in this group of patients and discuss advances in imaging that might improve our capabilities in the future.


Vibrational spectroscopic determination of local solvent electric field, solute-solvent electrostatic interaction energy, and their fluctuation amplitudes.
IR probes have been extensively used to monitor local electrostatic and solvation dynamics. Particularly, their vibrational frequencies are highly sensitive to local solvent electric field around an IR probe. Here, we show that the experimentally measured vibrational frequency shifts can be inversely used to determine local electric potential distribution and solute-solvent electrostatic interaction energy. In addition, the upper limits of their fluctuation amplitudes are estimated by using the vibrational bandwidths. Applying this method to fully deuterated N-methylacetamide (NMA) in D(2)O and examining the solvatochromic effects on the amide I' and II' mode frequencies, we found that the solvent electric potential difference between O(═C) and D(-N) atoms of the peptide bond is about 5.4 V, and thus, the approximate solvent electric field produced by surrounding water molecules on the NMA is 172 MV/cm on average if the molecular geometry is taken into account. The solute-solvent electrostatic interaction energy is estimated to be -137 kJ/mol, by considering electric dipole-electric field interaction. Furthermore, their root-mean-square fluctuation amplitudes are as large as 1.6 V, 52 MV/cm, and 41 kJ/mol, respectively. We found that the water electric potential on a peptide bond is spatially nonhomogeneous and that the fluctuation in the electrostatic peptide-water interaction energy is about 10 times larger than the thermal energy at room temperature. This indicates that the peptide-solvent interactions are indeed important for the activation of chemical reactions in aqueous solution.


Functional Neurocircuits and Neuroimaging Biomarkers of Tobacco Use Disorder.
Drug abuse and addiction remain major public health issues, exemplified by the opioid epidemic currently devastating the United States. Treatment outcomes across substance use disorders remain unacceptably poor, wherein drug discovery/development for this multifaceted neuropsychiatric disorder focuses on single molecular-level targets. Rather, our opinion is that a systems-level neuroimaging perspective is crucial for identifying novel therapeutic targets, biomarkers to stratify patients, and individualized treatment strategies. Focusing on tobacco use disorder, we advocate a brain systems-level perspective linking two abuse-related facets (i.e., statelike withdrawal and traitlike addiction severity) with specific neurocircuitry (insula- and striatum-centered networks). To the extent that precise neurocircuits mediate distinct facets of abuse, treatment development must adopt not only a systems-level perspective, but also multi-intervention rather than mono-intervention practices.


Structure and validity of Family Harmony Scale: An instrument for measuring harmony.
Culture plays a role in mental health, partly by defining the characteristics that are indicative of positive adjustment. In Chinese cultures, positive family relationships are considered central to well-being. The culturally emphasized characteristic of family harmony may be an important factor associated with psychopathology. This article presents the development and psychometric examination of the Family Harmony Scale (FHS), an indigenously developed 24-item instrument tapping family harmony in 17,461 Hong Kong residents from 7,791 households. A higher-order model with 1 second-order factor and 5 first-order factors fit the data well and showed factorial invariance across sex and participants in different family roles. A 5-item short form (FHS-5) was also developed, with 1 item from each first-order factor. The short scale showed, as expected, a single-factor structure with good fit. Both scales demonstrated high internal consistency, acceptable test-retest reliability, and good convergent and discriminant validity. The 24-item FHS was negatively associated with depressive symptoms after accounting for individual risk factors and general family function. Family harmony moderated the relationship between life stress and depressive symptoms such that those individuals who reported low family harmony had stronger associations between life stress and depressive symptoms. This study adds to the literature a systematically developed, multidimensional measure of family harmony, which may be an important psychological protective factor, in a large urban Chinese sample. The FHS-5 minimizes operational and respondent burdens, making it an attractive tool for large-scale epidemiological studies with Chinese populations in urban settings, where over half of China's 1.4 billion people reside.


Management of treatment-related symptoms in patients with breast cancer.
Although the benefits of current treatment strategies are well established, many cancer survivors are at risk for developing physiologic and psychological late effects of cancer treatment that might lead to premature mortality and morbidity and compromise their quality of life. Psychological symptoms include anxiety, depression, fatigue, difficulty sleeping, and loss of self-esteem. Physiologic symptoms include pain, numbness, cognitive impairment, weight gain, loss of sexual interest, spontaneous menopause, and peripheral neuropathy. Both length and quality of survival are important end points. The goal of this review is to summarize the psychological and physiologic symptoms related to breast cancer treatment; the prevalence, contributing therapies, and inter-relatedness of these symptoms; current interventions to prevent, ameliorate, or treat these symptoms; and effectiveness and safety of these interventions. The results of this review will identify the gaps in knowledge and assist in the design of assessments and approaches to improve mortality and quality of life and provide the foundation for the development of evidence-based guidelines to standardize palliative care in cancer survivors.


A novel "signal-on" photoelectrochemical sensor for ultrasensitive detection of alkaline phosphatase activity based on a TiO2/g-C3N4 heterojunction.
The use of alkaline phosphatase (ALP) as a biomarker in some diseases including hepatitis, obstructive jaundice, osteoblastic bone cancer, and osteomalacia is important in clinical diagnosis. Furthermore, ALP activity detection is an essential hot topic in environmental monitoring, biomedical research, and other research fields. In this study, a novel "signal-on" photoelectrochemical (PEC) biosensor based on the ALP-catalyzed phosphorylation reaction was designed to sensitively detect ALP activity. In this design, ascorbic acid-an electron donor-was catalytically produced by ALP from l-ascorbic acid 2-phosphate trisodium salt in situ, which results in an increased photocurrent response signal. For immobilizing the ALP on the electrode surface, poly diallyl dimethyl ammonium chloride was used for the conjugation of ALP, and titanium dioxide (TiO2)-a photoactive material-and graphite-like carbon nitride (g-C3N4) nanocomposites were prepared and characterized. TiO2 attached on g-C3N4 plays an important role for the biosensing purpose due to their good biocompatibility and chemical/thermal stability, while g-C3N4 provides the PEC response signal. Furthermore, the prepared TiO2/g-C3N4 nanocomposites can effectively suppress electron-hole recombinations, improve the excitation conversion efficiency, and make the best use of solar energy. The PEC biosensor for ALP activity detection displays a detection limit of 0.03 U L-1 (S/N = 3), which offers a new route for the ALP activity assay in human serum samples.


Strabismus Recognition Using Eye-Tracking Data and Convolutional Neural Networks.
Strabismus is one of the most common vision diseases that would cause amblyopia and even permanent vision loss. Timely diagnosis is crucial for well treating strabismus. In contrast to manual diagnosis, automatic recognition can significantly reduce labor cost and increase diagnosis efficiency. In this paper, we propose to recognize strabismus using eye-tracking data and convolutional neural networks. In particular, an eye tracker is first exploited to record a subject's eye movements. A gaze deviation (GaDe) image is then proposed to characterize the subject's eye-tracking data according to the accuracies of gaze points. The GaDe image is fed to a convolutional neural network (CNN) that has been trained on a large image database called ImageNet. The outputs of the full connection layers of the CNN are used as the GaDe image's features for strabismus recognition. A dataset containing eye-tracking data of both strabismic subjects and normal subjects is established for experiments. Experimental results demonstrate that the natural image features can be well transferred to represent eye-tracking data, and strabismus can be effectively recognized by our proposed method.


Prefrontal cortical response to conflict during semantic and phonological tasks.
Debates about the function of the prefrontal cortex are as old as the field of neuropsychology--often dated to Paul Broca's seminal work. Theories of the functional organization of the prefrontal cortex can be roughly divided into those that describe organization by process and those that describe organization by material. Recent studies of the function of the posterior, left inferior frontal gyrus (pLIFG) have yielded two quite different interpretations: One hypothesis holds that the pLIFG plays a domain-specific role in phonological processing, whereas another hypothesis describes a more general function of the pLIFG in cognitive control. In the current study, we distinguish effects of increasing cognitive control demands from effects of phonological processing. The results support the hypothesized role for the pLIFG in cognitive control, and more task-specific roles for posterior areas in phonology and semantics. Thus, these results suggest an alternative explanation of previously reported phonology-specific effects in the pLIFG.


Antimicrobial activities of amine- and guanidine-functionalized cholic acid derivatives.
Compounds in a series of cholic acid derivatives, designed to mimic the activities of polymyxin B and its derivatives, act as both potent antibiotics and effective permeabilizers of the outer membranes of gram-negative bacteria. Some of these compounds rival polymyxin B in antibacterial activity against gram-negative bacteria and are also very active against gram-positive organisms. Other compounds interact synergistically with hydrophobic antibiotics to inhibit bacterial growth.


Establishment and characterization of a panel of human uveal melanoma xenografts derived from primary and/or metastatic tumors.
Uveal melanoma is the most common primary intraocular malignant tumor in adults and is defined by a poor natural outcome, as 50% of patients die from metastases. The aim of this study was to develop and characterize a panel of human uveal melanoma xenografts transplanted into immunodeficient mice. Ninety tumor specimens were grafted into severe combined immunodeficient mice, and 25 transplantable xenografts were then established (28%). Relationship between tumor graft and clinical, biological, and therapeutic features of the patients included were investigated. Characterization of 16 xenografts included histology, molecular analyses by immunohistochemistry, genetic alteration analysis (single-nucleotide polymorphism), and specific tumor antigen expression by quantitative reverse transcription-PCR. Pharmacologic characterization (chemosensitivity) was also done in four models using two drugs, temozolomide and fotemustine, currently used in the clinical management of uveal melanoma. Take rate of human uveal melanoma was 28% (25 of 90). Tumor take was independent of size, histologic parameters, or chromosome 3 monosomy but was significantly higher in metastatic tumors. Interestingly, in vivo tumor growth was prognostic for a lower metastasis-free survival in patients with primary tumors. A high concordance between the patients' tumors and their corresponding xenografts was found for all parameters tested (histology, genetic profile, and tumor antigen expression). Finally, the four xenografts studied displayed different response profiles to chemotherapeutic agents. Based on these results, this panel of 16 uveal melanoma xenografts represents a useful preclinical tool for both pharmacologic and biological assessments.


Effect of Soil Moisture Regimes on Growth and Seed Production of Two Australian Biotypes of Sisymbrium thellungii O. E. Schulz.
Sisymbrium thellungii O. E. Schulz is an emerging problematic weed in the northern grain region of Australia. Several different biotypes exist in this region but not all biotypes exhibit the same growth and reproduction behavior. This might be due to local adaptation to the different agro-ecosystems, however, information on this aspect is limited. To determine whether adaptation to water stress was a factor in biotype demographic growth and reproduction behavior, we evaluated the physiological and biochemical responses of two Australian S. thellungii biotypes, selected from high (Dalby) and medium (St. George) rainfall areas, to different pot soil moisture levels corresponding to 100, 75, 50, and 25% of soil water holding capacity (WHC). Averaged across moisture levels, the St. George biotype (medium rainfall area) had 89% greater biomass and produced 321% more seeds than the Dalby biotype. The St. George biotype was less affected by increased levels of water stress than the Dalby biotype. The Dalby biotype produced 4,787 seeds plant-1 at 100% WHC and only 28 seeds plant-1 at 25% WHC. On the other hand, the St. George biotype produced 4,061 seeds plant-1 at 25% WHC and its seed production at 100% WHC was 9,834 seeds plant-1. On a per leaf area basis and averaged across all moisture levels, the St. George had significantly lower net carbon assimilation compared with the Dalby biotype, accompanied by a trend for lower stomatal conductance, which might indicate an adaptation to water stress. Across the moisture levels, the St. George biotype had higher phenolics and total soluble sugar, but free proline content was higher in the Dalby biotype compared with the St. George biotype. Like total soluble sugar, proline content increased with water stress in both biotypes, but it increased to a greater extent in the Dalby biotype, particularly at the 25% of WHC. Branching, flowering and maturity occurred earlier in the St. George biotype compared with the Dalby biotype, indicating relatively faster growth of the St. George biotype, which again seems to be an adaptation to water-limited environments. In conclusion, the St. George biotype of S. thellungii had higher reproductive capacity than the Dalby biotype across all the moisture regimes, which suggests greater invasiveness. Overall, the large size and rapid growth of the S. thellungii population from the medium rainfall area, together with its physiological response to water stress and its ability to maintain seed production in dry conditions, may enable this biotype to become widespread in Australia.


First Report of Powdery Mildew (Oidium sp.) on Akebia quinata in Italy.
Akebia quinata Decne., an ornamental species belonging to the family Lardizabalaceae, is used as a climbing species in gardens to cover walls as well as supports and is very much appreciated because of its dark red flowers. During the summer of 2003, severe outbreaks of a previously unknown powdery mildew were observed on established plantings in several gardens near Biella (northern Italy). The upper surfaces of leaves were covered with white mycelium, and the corresponding abaxial surface of infected leaves were chlorotic. Young, green stems also affected showed extended chlorosis. As the disease progressed, infected leaves turned yellow and died. Foot cell was cylindric and appressorium lobed. Conidia formed singly were hyaline, ellipsoid, and measured 26.4 to 45.6 × 10.6 to 15.6 μm (average 35.1 × 12.7 μm). Fibrosin bodies were not present. The pathogen was identified as Oidium sp. subgenus Pseudoidium (1) partially because cleistothecia were not observed. Conidial measurements are close to those reported for Microsphaera akebiae Sawada. Pathogenicity was confirmed by gently pressing diseased leaves onto leaves of healthy A. quinata plants. Three plants of A. quinata were used as replicates. Noninoculated plants served as controls. Plants were maintained between 20 and 30°C in a garden located 5 km from where the disease was originally found. After 10 days, typical symptoms of powdery mildew developed on inoculated plants. Noninoculated plants did not show symptoms. To our knowledge, this is the first report of the presence of powdery mildew on A. quinata in Italy. The disease is currently restricted to the area of Biella. The presence of M. akebiae was recently reported in the Netherlands (2). Specimens of this disease are available at the DIVAPRA Collection at the University of Torino. References: (1) U. Braun and S. Takamatsu. Schlechtendalia, 4:1, 2000. (2) M. Scholler and W. Gams. Nova Hedwigia, 67:101, 1998.


Heterotopic ossification revisited: a 21-year surgical experience.
Heterotopic ossification (HO) is an infrequently encountered complication of a burn. A retrospective review was undertaken to evaluate our treatment and results. Forty-two patients were identified with HO during 21 yrs. Mean age was 38 yrs. Mean total body surface area and third-degree burn were 55% and 37%, respectively. The elbow was the most frequent site (>90%), and 44% were bilateral. The next most common sites were shoulder, hip, knee, and forearm. Greater than 90% of patients had ventilator support and intensive care unit length of stay 58 and 79 days, respectively. HO was first suspected by decreased range of motion, painful and/or swollen joint, or a nerve deficit. Conventional radiographs were used to confirm the clinical diagnosis. The majority of burns overlying joints with HO were associated with prolonged wound closure because of depth, wound infection, or graft loss. Mean day of diagnosis was 71 days (range, 21-134). Excision of HO was undertaken only when range of motion compromised activities of daily living. Surgery successfully improved range of motion in all cases. The mean elbow arc of motion before and after surgery was 52 degrees and 119 degrees (range, 30-180 degrees), respectively. Seventy percent of elbows were ankylosed. A continuous passive motion device was instituted immediately postoperatively. Local postoperative complications included hematoma, wound dehiscence, infection, and nerve deficit. Maintaining range of motion was difficult for 75% of patients. Symptomatic recurrence of HO occurred in four elbows and one forearm. Because the cause(s) are unknown, prevention is impossible; once diagnosed, medical treatment is problematic and spontaneous resolution is infrequent. Surgery continues to be the recommended treatment when activities of daily living or life style are affected.


Obesity, diabetes, and associated costs of exposure to endocrine-disrupting chemicals in the European Union.
Obesity and diabetes are epidemic in the European Union (EU). Exposure to endocrine-disrupting chemicals (EDCs) is increasingly recognized as a contributor, independent of diet and physical activity. The objective was to estimate obesity, diabetes, and associated costs that can be reasonably attributed to EDC exposures in the EU. An expert panel evaluated evidence for probability of causation using weight-of-evidence characterization adapted from that applied by the Intergovernmental Panel on Climate Change. Exposure-response relationships and reference levels were evaluated for relevant EDCs, and biomarker data were organized from peer-reviewed studies to represent European exposure and burden of disease. Cost estimation as of 2010 utilized published cost estimates for childhood obesity, adult obesity, and adult diabetes. Setting, Patients and Participants, and Intervention: Cost estimation was performed from the societal perspective. The panel identified a 40% to 69% probability of dichlorodiphenyldichloroethylene causing 1555 cases of overweight at age 10 (sensitivity analysis: 1555-5463) in 2010 with associated costs of €24.6 million (sensitivity analysis: €24.6-86.4 million). A 20% to 39% probability was identified for dichlorodiphenyldichloroethylene causing 28 200 cases of adult diabetes (sensitivity analysis: 28 200-56 400) with associated costs of €835 million (sensitivity analysis: €835 million-16.6 billion). The panel also identified a 40% to 69% probability of phthalate exposure causing 53 900 cases of obesity in older women and €15.6 billion in associated costs. Phthalate exposure was also found to have a 40% to 69% probability of causing 20 500 new-onset cases of diabetes in older women with €607 million in associated costs. Prenatal bisphenol A exposure was identified to have a 20% to 69% probability of causing 42 400 cases of childhood obesity, with associated lifetime costs of €1.54 billion. EDC exposures in the EU contribute substantially to obesity and diabetes, with a moderate probability of >€18 billion costs per year. This is a conservative estimate; the results emphasize the need to control EDC exposures.


Identification of the gene coding for rhesus cytomegalovirus glycoprotein B and immunological analysis of the protein.
The nucleotide sequence of the gene encoding glycoprotein B (gB) of rhesus cytomegalovirus (RhCMV) was determined and the protein characterized. The open reading frame of gB encoded a protein of 854 amino acids with 60% identity and 75% similarity at the amino acid level to human cytomegalovirus (HCMV) gB. Cysteine residues in the extraluminal part of the protein are perfectly conserved. Out of the 16 potential N-linked glycosylation sites present in HCMV gB, 15 are conserved in RhCMV gB. Immunoblot analyses with antisera detected three bands of 150 kDa, 90-110 kDa and 55 kDa representing the full-length gB as well as the proteolytic cleavage products. Cross-reactivity and cross-neutralization of a number of HCMV gB-specific monoclonal antibodies with RhCMV gB indicated sharing of immunogenic epitopes between the two molecules. The RhCMV gB regions corresponding to antigenic domains AD-1, 2 and 3 of HCMV gB were immunogenic during natural RhCMV infection with the AD-1 region being the immunodominant domain. The data indicate that RhCMV might represent a useful model to investigate pathogenesis and immune surveillance of cytomegaloviruses.


Enhanced muscle glucose metabolism after exercise in the rat: the two phases.
Thirty minutes after a treadmill run, glucose utilization and glycogen synthesis in perfused rat skeletal muscle are enhanced due to an increase in insulin sensitivity (Richter et al., J. Clin. Invest. 69: 785-793, 1982). The exercise used in these studies was of moderate intensity, and muscle glycogen was substantially repleted at the time (30 min postexercise) that glucose metabolism was examined. When rats were run at twice the previous rate (36 m/min), muscle glycogen was still substantially diminished 30 min after the run. At this time the previously noted increase in insulin sensitivity was still observed in perfused muscle; however, glucose utilization was also increased in the absence of added insulin (1.5 vs. 4.2 mumol X g-1 X h-1). In contrast 2.5 h after the run, muscle glycogen had returned to near preexercise values, and only the insulin-induced increase in glucose utilization was evident. The data suggest that the restoration of muscle glycogen after exercise occurs in two phases. In phase I, muscle glycogen is depleted and insulin-stimulated glucose utilization and glucose utilization in the absence of added insulin may both be enhanced. In phase II glycogen levels have returned to near base-line values and only the increase in insulin sensitivity persists. It is proposed that phase I corresponds to the period of rapid glycogen repletion that immediately follows exercise and phase II to the period of supercompensation.


Upregulation of KCa3.1 K(+) channel in mesenteric lymph node CD4(+) T lymphocytes from a mouse model of dextran sodium sulfate-induced inflammatory bowel disease.
The intermediate-conductance Ca(2+)-activated K(+) channel KCa3.1/KCNN4 plays an important role in the modulation of Ca(2+) signaling through the control of the membrane potential in T lymphocytes. Here, we study the involvement of KCa3.1 in the enlargement of the mesenteric lymph nodes (MLNs) in a mouse model of inflammatory bowel disease (IBD). The mouse model of IBD was prepared by exposing male C57BL/6J mice to 5% dextran sulfate sodium for 7 days. Inflammation-induced changes in KCa3.1 activity and the expressions of KCa3.1 and its regulators in MLN CD4(+) T lymphocytes were monitored by real-time PCR, Western blot, voltage-sensitive dye imaging, patch-clamp, and flow cytometric analyses. Concomitant with an upregulation of KCa3.1a and nucleoside diphosphate kinase B (NDPK-B), a positive KCa3.1 regulator, an increase in KCa3.1 activity was observed in MLN CD4(+) T lymphocytes in the IBD model. Pharmacological blockade of KCa3.1 elicited the following results: 1) a significant decrease in IBD disease severity, as assessed by diarrhea, visible fecal blood, inflammation, and crypt damage of the colon and MLN enlargement compared with control mice, and 2) the restoration of the expression levels of KCa3.1a, NDPK-B, and Th1 cytokines in IBD model MLN CD4(+) T lymphocytes. These findings suggest that the increase in KCa3.1 activity induced by the upregulation of KCa3.1a and NDPK-B may be involved in the pathogenesis of IBD by mediating the enhancement of the proliferative response in MLN CD4(+) T lymphocyte and, therefore, that the pharmacological blockade of KCa3.1 may decrease the risk of IBD.


Role of plant stomata in bacterial invasion.
Stomata are microscopic pores in the epidermis of the aerial parts of terrestrial plants. These pores are essential for photosynthesis, as they allow CO(2) to diffuse into the plant. The size of the stomatal pore changes in response to environmental conditions, such as light intensity, air humidity and CO(2) concentrations, as part of the plant's adaptation to maximize photosynthetic efficiency and, at the same time, to minimize water loss. Historically, stomata have been considered as passive portal of entry for plant pathogenic bacteria. However, recent studies suggest that stomata can play an active role in restricting bacterial invasion as part of the plant innate immune system. Some plant pathogens have evolved specific virulence factors to overcome stomata-based defence. Interestingly, many bacterial disease outbreaks require high humidity, rain, or frost damage, which could promote stomatal opening and/or bypass stomatal defence by creating wounds as alternative entry sites. Further studies on microbial and environmental regulation of stomata-based defence should fill gaps in our understanding of bacterial pathogenesis, disease epidemiology and phyllosphere microbiology.


Heterogeneity of clara cell glutathione. A possible basis for differences in cellular responses to pulmonary cytotoxicants.
Clara-cell populations show a high degree of variation in susceptibility to injury by bioactivated cytotoxicants. Because glutathione (GSH) is critical for detoxification of electrophilic metabolites, heterogeneity in Clara cell GSH levels may lead to a wide range of cytotoxic responses. This study was designed to define the distinct GSH pools within Clara cells, characterize heterogeneity within the population, and examine whether heterogeneity contributes to susceptibility. Using fluorescent imaging combined with high-performance liquid chromatography analysis, semiquantitative measurements were obtained by evaluation of GSH using monochlorobimane and monobromobimane. In steady-state conditions, the GSH measured in isolated cells was in the femtomole range, but varied 4-fold between individual cells. Clara cells analyzed in situ and in vitro confirmed this heterogeneity. The response of these cells to compounds that modulate GSH was also variable. Diethylmaleate depleted GSH, whereas GSH monoethylester augmented it. However, both acted nonuniformly in isolated Clara cells. The depletion of intracellular GSH caused a striking decrease in cell viability upon incubation with naphthalene (NA). The sulfhydryl-binding fluorochrome BODIPY, which colocalized with tetramethylrosamine, a mitochondrial dye, demonstrated by confocal microscopy that cellular sulfhydryls are highest in the mitochondria, next-highest in cytoplasm, and lowest in the nucleus. These pools responded differently to modulators of GSH. We concluded that the steady-state intracellular GSH of Clara cells exists in distinct pools and is highly heterogeneous within the population, and that the heterogeneity of GSH levels corresponds closely to the response of Clara cells to injury by NA.


Biochar Aging: Mechanisms, Physicochemical Changes, Assessment, And Implications for Field Applications.
Biochar has triggered a black gold rush in environmental studies as a carbon-rich material with well-developed porous structure and tunable functionality. While much attention has been placed on its apparent ability to store carbon in the ground, immobilize soil pollutants, and improve soil fertility, its temporally evolving in situ performance in these roles must not be overlooked. After field application, various environmental factors, such as temperature variations, precipitation events and microbial activities, can lead to its fragmentation, dissolution, and oxidation, thus causing drastic changes to the physicochemical properties. Direct monitoring of biochar-amended soils can provide good evidence of its temporal evolution, but this requires long-term field trials. Various artificial aging methods, such as chemical oxidation, wet-dry cycling and mineral modification, have therefore been designed to mimic natural aging mechanisms. Here we evaluate the science of biochar aging, critically summarize aging-induced changes to biochar properties, and offer a state-of-the-art for artificial aging simulation approaches. In addition, the implications of biochar aging are also considered regarding its potential development and deployment as a soil amendment. We suggest that for improved simulation and prediction, artificial aging methods must shift from qualitative to quantitative approaches. Furthermore, artificial preaging may serve to synthesize engineered biochars for green and sustainable environmental applications.


Effects of Extended Postmortem Interval on Microbial Communities in Organs of the Human Cadaver.
Human thanatomicrobiota studies have shown that microorganisms inhabit and proliferate externally and internally throughout the body and are the primary mediators of putrefaction after death. Yet little is known about the source and diversity of the thanatomicrobiome or the underlying factors leading to delayed decomposition exhibited by reproductive organs. The use of the V4 hypervariable region of bacterial 16S rRNA gene sequences for taxonomic classification ("barcoding") and phylogenetic analyses of human postmortem microbiota has recently emerged as a possible tool in forensic microbiology. The goal of this study was to apply a 16S rRNA barcoding approach to investigate variation among different organs, as well as the extent to which microbial associations among different body organs in human cadavers can be used to predict forensically important determinations, such as cause and time of death. We assessed microbiota of organ tissues including brain, heart, liver, spleen, prostate, and uterus collected at autopsy from criminal casework of 40 Italian cadavers with times of death ranging from 24 to 432 h. Both the uterus and prostate had a significantly higher alpha diversity compared to other anatomical sites, and exhibited a significantly different microbial community composition from non-reproductive organs, which we found to be dominated by the bacterial orders MLE1-12, Saprospirales, and Burkholderiales. In contrast, reproductive organs were dominated by Clostridiales, Lactobacillales, and showed a marked decrease in relative abundance of MLE1-12. These results provide insight into the observation that the uterus and prostate are the last internal organs to decay during human decomposition. We conclude that distinct community profiles of reproductive versus non-reproductive organs may help guide the application of forensic microbiology tools to investigations of human cadavers.


Intracellular transport of viruses and their components: utilizing the cytoskeleton and membrane highways.
Plant viruses are obligate organisms that require host components for movement within and between cells. A mechanistic understanding of virus movement will allow the identification of new methods to control virus systemic spread and serve as a model system for understanding host macromolecule intra- and intercellular transport. Recent studies have moved beyond the identification of virus proteins involved in virus movement and their effect on plasmodesmal size exclusion limits to the analysis of their interactions with host components to allow movement within and between cells. It is clear that individual virus proteins and replication complexes associate with and, in some cases, traffic along the host cytoskeleton and membranes. Here, we review these recent findings, highlighting the diverse associations observed between these components and their trafficking capacity. Plant viruses operate individually, sometimes within virus species, to utilize unique interactions between their proteins or complexes and individual host cytoskeletal or membrane elements over time or space for their movement. However, there is not sufficient information for any plant virus to create a complete model of its intracellular movement; thus, more research is needed to achieve that goal.


Effect of body mass index on in vitro fertilization outcomes in women.
Obesity has become a major health problem across the world. In women, it is known to cause anovulation, subfecundity, increased risk of fetal anomalies and miscarriage rates. However, in women going for assisted reproduction the effects of obesity on egg quality, embryo quality, clinical pregnancy, live birth rates are controversial. To assess the effect of women's body mass index (BMI) on the reproductive outcome of non donor In vitro fertilization (IVF)/Intracytoplasmic sperm injection (ICSI). The effects of BMI on their gonadotrophin levels (day 2 LH, FSH), gonadotrophin dose required for ovarian stimulation, endometrial thickness and oocyte/embryo quality were looked at, after correcting for age and poor ovarian reserve. Retrospective study of medical records of 308 women undergoing non donor IVF cycles in a University affiliated teaching hospital. They were classified into three groups: normal weight (BMI<25 kg/m(2)), overweight (BMI>25 <30 kg/m(2)) and obese (BMI>30 kg/m(2)). All women underwent controlled ovarian hyper stimulation using long agonist protocol. There were 88 (28.6%) in the normal weight group, 147 (47.7%) in the overweight and 73 (23.7%) in the obese group. All three groups were comparable with respect to age, duration of infertility, female and male causes of infertility. The three groups were similar with respect to day 2 LH/FSH levels, endometrial thickness and gonadotrophin requirements, oocyte quality, fertilization, cleavage rates, number of good quality embryos and clinical pregnancy rates. Increase in body mass index in women does not appear to have an adverse effect on IVF outcome. However, preconceptual counselling for obese women is a must as weight reduction helps in reducing pregnancy-related complications.


Forest work and its implications for malaria elimination: a qualitative study.
Over the last 20 years, malaria incidence has decreased across the Greater Mekong Sub-region (GMS) and the emergence of artemisinin resistance has stimulated efforts to accelerate regional elimination. In the GMS, the malaria transmission is focused increasingly in forested zones. This article describes forest-going activities and examines forest workers' attitudes to and experiences of malaria prevention and control in north-eastern Cambodia. In Stung Treng Province, Cambodia, 19 in-depth interviews were conducted in villages with participants recently diagnosed with uncomplicated falciparum malaria who reported working in forests. Two focus group discussions with respondents' forest-working peers were held. Interviews and focus groups were audio-recorded transcribed, and translated for thematic analysis. Forest work is an essential source of income for respondents. Many combine it with farming, which influences the timing and duration of forest visits. Forest activities include logging and collecting other forest products, particularly malva nuts. Men log year-round, whereas gathering forest products is seasonal and can involve entire families. Forest workers sleep chiefly in unimpregnated hammock nets in make-shift encampments. Respondents are concerned about symptomatic malaria, but unfamiliar with the concept of asymptomatic infection. They view the forest as an area of potential malaria infection and seek to protect themselves from mosquito bites through wearing long-sleeved clothes, using repellents, and lighting fires. Forest workers express a willingness to self-test and self-administer anti-malarials. Forest workers' behaviour and perceptions of risk indicate that improvements are needed to current control measures. There is potential to: better target distribution of impregnated hammock nets; offer curative or presumptive treatment while in forests; and expand access to screening. Establishing the efficacy and feasibility of prophylaxis for forest workers in the GMS is a priority.


Paralytic Shellfish Toxins Occurrence in Non-Traditional Invertebrate Vectors from North Atlantic Waters (Azores, Madeira, and Morocco).
Paralytic shellfish toxins (PSTs) are potent alkaloids of microalgal and cyanobacterial origin, with worldwide distribution. Over the last 20 years, the number of poisoning incidents has declined as a result of the implementation of legislation and monitoring programs based on bivalves. In the summer of 2012 and 2013, we collected a total of 98 samples from 23 different species belonging to benthic and subtidal organisms, such as echinoderms, crustaceans, bivalves, and gastropods. The sampling locations were Madeira, São Miguel Island (Azores archipelago), and the northwestern coast of Morocco. The samples were analyzed using post-column oxidation liquid chromatography with a fluorescence detection method. Our main goal was to detect new vectors for these biotoxins. After reporting a total of 59 positive results for PSTs with 14 new vectors identified, we verified that some of the amounts exceeded the limit value established in the EU. These results suggest that routine monitoring of saxitoxin and its analogs should be extended to more potential vectors other than bivalves, including other edible organisms, for a better protection of public health.


Electrostatic potential surface analysis of the transition state for AMP nucleosidase and for formycin 5'-phosphate, a transition-state inhibitor.
AMP nucleosidase hydrolyzes the N-glycosidic bond of AMP to yield adenine and ribose 5-phosphate. Kinetic isotope effects have been used to establish an experimentally based transition-state structure for the native enzyme and a Vmax mutant [Mentch, F., Parkin, D. W., & Schramm, V. L. (1987) Biochemistry 26, 921-930; Parkin, D. W., Mentch, F., Banks, G. A., Horenstein, B. A., & Schramm, V. L. (1991) Biochemistry 30, 4586-4594]. The transition states are characterized by weak reaction coordinate bonds to C1' and substantial carbocation character in the ribose ring. The N9-C1' bond to the leaving group is nearly broken and the adenine ring is protonated at the transition state. Formycin 5'-phosphate and other purine nucleoside 5'-phosphate analogues with syn-glycosyl torsion angles bind better than substrate, supporting a syn configuration in the enzyme-substrate complex and presumably in the transition state [Giranda, V. L., Berman, H. M., & Schramm, V. L. (1988) Biochemistry 27, 5813-5818]. Access to a geometric model of the transition state permits the analysis of its molecular electrostatic potential surface as enforced by the enzyme. Comparison of the molecular electrostatic potential surfaces for AMP, formycin 5'-phosphate, and the transition state reveals a striking similarity in the surface charges of formycin 5'-phosphate and the transition state. The enzyme-stabilized transition state for AMP hydrolysis is characterized by new positive electrostatic potential in the adenine ring as a result of protonation by the enzyme. This is closely matched by the protonated pyrazole ring of formycin 5'-phosphate. The molecular electrostatic potential surfaces of formycin 5'-phosphate and the transition state for AMP are similar and are likely to be a factor in the Km/Ki value of > 10(3) for formycin 5'-phosphate as a transition-state inhibitor of AMP nucleosidase.


Sense of coherence and 22-year all-cause mortality in adult men.
Sense of coherence (SOC) is a central construct in Antonovsky's salutogenic theory, which focuses on people's health-promoting and health-protecting characteristics. We examined prospectively the association of SOC with all-cause mortality during 22 years (1989-2011). The data of 585 men from the Israel longitudinal study of Glucose Intolerance, Obesity, and Hypertension (The Israel GOH) comprised the analytic sample. Participants were 48-67 years old at study entry (1989). Information on sociodemographic, medical history and health-related risk factors were obtained at baseline through a face-to-face interview. Participants completed Antonovsky's 29-item SOC scale. Information on all-cause mortality was obtained from the Israeli Mortality Register (1989 through 2011). We evaluated the effect of SOC on time-to-death using multiple Cox proportional hazard regression. Controlling for sociodemographic, smoking status and morbidities, there was strong evidence of an association between SOC and 22-year all-cause mortality [adjusted hazard ratio (aHR)=0.992, 95% CI 0.986-0.998 per unit]. Strong SOC was associated with a 35% reduction in all-cause mortality relative to weak SOC (aHR=0.653, 95% CI 0.454-0.939). There was no convincing evidence of a survival advantage for individuals with intermediate level of SOC relative to persons reporting weak SOC (aHR=0.821, 95% CI 0.595-1.134). Our study provides strong evidence of an association between SOC and mortality, above and beyond potential confounding factors and established risk factors. Considerable more research is needed on the role of SOC in health and survival and the potential pathways linking SOC and health.


Penicillium purpurogenum Produces a Set of Endoxylanases: Identification, Heterologous Expression, and Characterization of a Fourth Xylanase, XynD, a Novel Enzyme Belonging to Glycoside Hydrolase Family 10.
The fungus Penicillium purpurogenum grows on a variety of natural carbon sources and secretes a large number of enzymes which degrade the polysaccharides present in lignocellulose. In this work, the gene coding for a novel endoxylanase has been identified in the genome of the fungus. This gene (xynd) possesses four introns. The cDNA has been expressed in Pichia pastoris and characterized. The enzyme, XynD, belongs to family 10 of the glycoside hydrolases. Mature XynD has a calculated molecular weight of 40,997. It consists of 387 amino acid residues with an N-terminal catalytic module, a linker rich in ser and thr residues, and a C-terminal family 1 carbohydrate-binding module. XynD shows the highest identity (97%) to a putative endoxylanase from Penicillium subrubescens but its highest identity to a biochemically characterized xylanase (XYND from Penicillium funiculosum) is only 68%. The enzyme has a temperature optimum of 60 °C, and it is highly stable in its pH optimum range of 6.5-8.5. XynD is the fourth biochemically characterized endoxylanase from P. purpurogenum, confirming the rich potential of this fungus for lignocellulose biodegradation. XynD, due to its wide pH optimum and stability, may be a useful enzyme in biotechnological procedures related to this biodegradation process.


Identification of bovine sperm head for morphometry analysis in quantitative phase-contrast holographic microscopy.
An investigation is reported of the identification and measurement of region of interest (ROI) in quantitative phase-contrast maps of biological cells by digital holographic microscopy. In particular, two different methods have been developed for in vitro bull sperm head morphometry analysis. We show that semen analysis can be accomplished by means of the proposed techniques . Extraction and measurement of various parameters are performed. It is demonstrated that both proposed methods are efficient to skim the data set in a preselective analysis for discarding anomalous data.


Optical changes in unilamellar vesicles experiencing osmotic stress.
Membrane properties that vary as a result of isotropic and transmembrane osmolality variations (osmotic stress) are of considerable relevance to mechanisms such as osmoregulation, in which a biological system "senses" and responds to changes in the osmotic environment. In this paper the light-scattering behavior of a model system consisting of large unilamellar vesicles of dioleoyl phosphatidyl glycerol (DOPG) is examined as a function of their osmotic environment. Osmotic downshifts lead to marked reductions in the scattered intensity, whereas osmotic upshifts lead to strong intensity increases. It is shown that these changes in the scattering intensity involve changes in the refractive index of the membrane bilayer that result from an alteration in the extent of hydration and/or the phospholipid packing density. By considering the energetics of osmotically stressed vesicles, and from explicit analysis of the Rayleigh-Gans-Debye scattering factors for spherical and ellipsoidal shells, we quantitatively demonstrate that although changes in vesicle volume and shape can arise in response to the imposition of osmotic stress, these factors alone cannot account for the observed changes in scattered intensity.


Variants in ZNRD1 gene predict HIV-1/AIDS disease progression in a Han Chinese population in Taiwan.
Patients demonstrate notable variations in disease progression following human immunodeficiency virus (HIV) infection. We aimed to identify ZNRD1 and RNF39 genetic variants linked to AIDS progression. We conducted a genetic association study in HIV-1-infected Han Chinese patients residing in Taiwan. The clinical characteristics of 143 HIV-1-infected patients were measured, and patients were split into 2 groups: AIDS progression and AIDS non-progression. Genotyping of ZNRD1 and RNF39 was performed in all participants. We found that patients in the AIDS progression group had higher HIV-1 viral loads and lower CD4 cell counts than did patients in the AIDS non-progression group. The frequency of the AA genotype of ZNRD1 (rs16896970) was lower in the AIDS progression group than in the AIDS non-progression group. Patients with AA genotypes had lower levels of HIV-1 viral loads and higher levels of CD4 cell counts than did patients with AG+GG genotypes. AIDS progression in patients with the AA group is significantly different from that in patients with the AG and GG groups by using Kaplan-Meier survival analysis. The hazard ratio for progression was lower in the AA group than in the AG and GG groups. We identified a SNP that contributes to AIDS progression in HIV-1-infected patients in this population. This SNP had a significant protective influence on AIDS progression, and polymorphisms of the ZNRD1 gene may play a role in the pathogenesis of HIV-1 infection.


Taxa-specific activity loss and mortality patterns in freshwater trematode cercariae under subarctic conditions.
Cercarial activity and survival are crucial traits for the transmission of trematodes. Temperature is particularly important, as faster depletion of limited cercarial energy reserves occurs at high temperatures. Seasonal climate conditions in high latitude regions may be challenging to complete trematode life cycle during the 6-month ice-free period, but temperature effects on the activity and survival of freshwater cercariae have not been previously identified. After experimentally simulating natural subarctic conditions during warmer and colder months (13 and 6°C), a statistical approach identifying changes in the tendency of cercarial activity loss and mortality data was used to detect differences in three trematode genera, represented by four taxa (Diplostomum spp., Apatemon spp., small- and large-sized Plagiorchis spp.). A strong temperature-dependent response was identified in both activity loss and mortality in all taxa, with Diplostomum spp. cercariae showing the most gradual changes compared to other taxa. Furthermore, whilst activity loss and mortality dynamics could not be divided into 'fish- vs invertebrate-infecting cercariae' groups, the detected taxa-specific responses in relation to life-history traits indicate the swimming behaviour of cercariae and energy allocation among larvae individuals as the main drivers. Cercariae exploit the short transmission window that allows a stable continuance of trematodes' life cycles in high-latitude freshwater ecosystems.


[Treatment of the extensive cicatricial stenoses of the upper third of trachea].
Treatment tactics in patients with the extensive cicatricial stenoses of the upper third of tracheа were thoroughly analyzed. Circular resections of trachea was performed in 51 cases, 18 of those were supplemented with the distal laryngeal resection. Combination of the endoscopic treatment (tracheal bouginage and laser stenosis dissection or tracheal recanalization) and circular resection of the trachea was performed in 23 patients. Endoscopic treatment modality proved to be the temporary measure by the extensive cicatricial stenoses or supratracheostomic obliteration of the trachea, which, nevertheless, permit and facilitates further radical circular tracheal resection.


The voice of the public in public health policy and planning: the role of public judgment.
Community involvement with public health planning and implementation are vital to improving community health. There are a variety of community health models that are available. We describe these four models from the perspective of how they involve the broader community. These models are evaluated from a different perspective about linking the community and politics and agencies, that involves naming issues, framing options, public deliberation and public acting. We suggest ways that these models can be further refined to connect citizens to the processes that we use for community health improvement.


Regulation of GLI1 by cis DNA elements and epigenetic marks.
GLI1 is one of three transcription factors (GLI1, GLI2 and GLI3) that mediate the Hedgehog signal transduction pathway and play important roles in normal development. GLI1 and GLI2 form a positive-feedback loop and function as human oncogenes. The mouse and human GLI1 genes have untranslated 5' exons and large introns 5' of the translational start. Here we show that Sonic Hedgehog (SHH) stimulates occupancy in the introns by H3K27ac, H3K4me3 and the histone reader protein BRD4. H3K27ac and H3K4me3 occupancy is not significantly changed by removing BRD4 from the human intron and transcription start site (TSS) region. We identified six GLI binding sites (GBS) in the first intron of the human GLI1 gene that are in regions of high sequence conservation among mammals. GLI1 and GLI2 bind all of the GBS in vitro. Elimination of GBS1 and 4 attenuates transcriptional activation by GLI1. Elimination of GBS1, 2, and 4 attenuates transcriptional activation by GLI2. Eliminating all sites essentially eliminates reporter gene activation. Further, GLI1 binds the histone variant H2A.Z. These results suggest that GLI1 and GLI2 can regulate GLI1 expression through protein-protein interactions involving complexes of transcription factors, histone variants, and reader proteins in the regulatory intron of the GLI1 gene. GLI1 acting in trans on the GLI1 intron provides a mechanism for GLI1 positive feedback and auto-regulation. Understanding the combinatorial protein landscape in this locus will be important to interrupting the GLI positive feedback loop and providing new therapeutic approaches to cancers associated with GLI1 overexpression.


Diet adaptation in dog reflects spread of prehistoric agriculture.
Adaptations allowing dogs to thrive on a diet rich in starch, including a significant AMY2B copy number gain, constituted a crucial step in the evolution of the dog from the wolf. It is however not clear whether this change was associated with the initial domestication, or represents a secondary shift related to the subsequent development of agriculture. Previous efforts to study this process were based on geographically limited data sets and low-resolution methods, and it is therefore not known to what extent the diet adaptations are universal among dogs and whether there are regional differences associated with alternative human subsistence strategies. Here we use droplet PCR to investigate worldwide AMY2B copy number diversity among indigenous as well as breed dogs and wolves to elucidate how a change in dog diet was associated with the domestication process and subsequent shifts in human subsistence. We find that AMY2B copy numbers are bimodally distributed with high copy numbers (median 2nAMY2B=11) in a majority of dogs but no, or few, duplications (median 2nAMY2B=3) in a small group of dogs originating mostly in Australia and the Arctic. We show that this pattern correlates geographically to the spread of prehistoric agriculture and conclude that the diet change may not have been associated with initial domestication but rather the subsequent development and spread of agriculture to most, but not all regions of the globe.


Cannabidiol as potential anticancer drug.
Over the past years, several lines of evidence support an antitumourigenic effect of cannabinoids including Δ(9)-tetrahydrocannabinol (Δ(9)-THC), synthetic agonists, endocannabinoids and endocannabinoid transport or degradation inhibitors. Indeed, cannabinoids possess anti-proliferative and pro-apoptotic effects and they are known to interfere with tumour neovascularization, cancer cell migration, adhesion, invasion and metastasization. However, the clinical use of Δ(9)-THC and additional cannabinoid agonists is often limited by their unwanted psychoactive side effects, and for this reason interest in non-psychoactive cannabinoid compounds with structural affinity for Δ(9)-THC, such as cannabidiol (CBD), has substantially increased in recent years. The present review will focus on the efficacy of CBD in the modulation of different steps of tumourigenesis in several types of cancer and highlights the importance of exploring CBD/CBD analogues as alternative therapeutic agents.


Postoperative magnetic resonance imaging findings after transtemporal and translabyrinthine vestibular schwannoma resection.
Magnetic resonance imaging (MRI) has become the investigation of choice to follow up patients after vestibular schwannoma resection. Retrospective. Postoperative MRI findings of 70 patients after vestibular schwannoma resection through a transtemporal (n = 48) and a translabyrinthine (n = 22) approach were reviewed. Time-dependent changes in intensity, size, and shape of enhancement in the internal auditory canal before and after contrast administration, postoperative temporal lobe gliosis, and changes of fat grafts were evaluated. After vestibular schwannoma resection, all patients showed signal enhancements in the internal auditory canal ranging from a faint to high signal intensity in the first postoperative MRI, 3 to 6 months after surgery. In the next MRI at 12 to 24 months after surgery, 30 patients (43%) showed a decreased signal, 35 patients (50%) a stable enhancement, and 5 patients (7%) an increased enhancement in the internal auditory canal depicted as an intense nodular or mass-like pattern. In patients with decreased or stable enhancement, a residual tumor could be excluded in the following MRI scans, whereas in all patients with increased enhancements after 12 to 24 months, signal enhancement further increased and residual tumors were detected. Different degrees of temporal lobe gliosis were found in 15 of 48 cases (31%) after transtemporal tumor removal. Enhancement of fat grafts used in 22 cases decreased to different degrees in 14 cases (64%). Differentiation of residual tumor from scar tissue in the internal auditory canal after vestibular schwannoma resection requires close, long-term follow-up. Nodular and progressive enhancements in the internal auditory canal indicate residual tumor. Linear enhancement in the internal auditory canal has been found to be a common finding after vestibular schwannoma resection not associated with residual tumor.


A multicenter comparative acute myeloid leukemia study: can we explain the differences in the outcomes in resource-constrained settings?
Outcomes in acute myeloid leukemia (AML) are dependent on patient- and disease-characteristics, treatment, and socioeconomic factors. AML outcomes between resource-constrained and developed countries have not been compared directly. We analyzed two cohorts: from São Paulo state, Brazil (USP, n = 312) and Oxford, United Kingdom (OUH, n = 158). USP cohort had inferior 5-year overall survival compared with OUH (29% vs. 49%, adjusted-p=.027). USP patients have higher early-mortality (23% vs. 6% p<.001) primarily due to multi-resistant Gram-negative bacterial and fungal infections. USP had higher 5-year cumulative incidence of relapse (60% vs. 50%, p=.0022), were less likely to undergo hematopoietic stem cell transplant (HSCT) (28% vs. 75%, p<.001) and waited longer for HSCT (median, 23.8 vs. 7.2 months, p<.001). Three-year survival in relapsed patients was worse in USP than OUH (10% vs. 39%, p<.001). Our study indicates that efforts to improve AML outcomes in Brazil should focus on infection prevention and control, and access to HSCT.


Evidence of Epstein-Barr virus association with gastric cancer and non-atrophic gastritis.
Different lines of evidence support an association between Epstein-Barr virus (EBV) and gastric cancer (GC). The main understood risk factor to develop GC is infection by Helicobacter pylori (H. pylori), which triggers a local inflammatory response critical for progression from gastritis to GC. The role of EBV in early inflammatory gastric lesions has been poorly studied. A recent study proposed a cutoff value of 2000 EBV particles to identify patients with increased chances of infection of the gastric epithelium, which may favor the inflammatory process. To better understand the role of EBV in cancer progression, we analyzed 75 samples of GC, 147 control samples of non-tumor gastric tissue derived from GC patients and 75 biopsies from patients with non-atrophic gastritis (NAG). A first-round PCR was used for EBV detection in tumor and non-tumor controls and a more sensitive nested PCR for gastritis samples; both PCRs had lower detection limits above the proposed cutoff value. With this strategy 10.67% of GC, 1.3% of non-tumor controls and 8% of gastritis samples were found positive. An EBER1 in situ hybridization showed EBV infection of epithelial cells in GC and in a third of NAG samples, while in the other NAGs infection was restricted to the mononuclear cell infiltrate. EBV-positive GCs were enriched in lace and cribriform patterns, while these rare patterns were not observed in EBV negative samples. Our results support a role for EBV in GC and early precursor lesions, either as directly oncogenic infecting epithelial cells or indirectly as an inflammatory trigger.


From the bedside to the bench and back again: predicting and improving the outcomes of SLT glaucoma therapy.
To determine whether selective laser trabeculoplasty (SLT) and prostaglandin analogues (PGAs) have a common mechanism of action that involves increasing conductivity across Schlemm's canal endothelial cells (SCEs) and inducing a similar decrease in intraocular pressure (IOP) in a given patient. The intercellular junctions in SCEs were made visible by transfection of a plasmid containing a GFP-tagged gene for ZO-1 protein. Transfected SCEs were treated with media conditioned by lasered trabecular meshwork endothelial cells (TMEs), or with latanoprost, bimatoprost, or travoprost. Non-transfected SCEs were exposed to brimonidine, timolol, or brinzolamide. Confocal microscopy and conductivity measurements documented the in vitro treatment effects. Clinically, the IOP in the first SLT-treated eye of 24 patients was measured (1) while on PGA therapy, (2) at "baseline" several weeks after discontinuing PGA therapy, and (3) approximately 90 days after SLT treatment. Both the in vitro addition of any of the 3 PGAs and of media conditioned by lasered TMEs induced similar SCE effects involving junction disassembly, paracellular pathway widening, and increased conductivity. Clinically, PGAs decreased IOP by a mean of 5.58 mmHg and SLT decreased IOP by 6.60 mmHg from a baseline of 21.52 mmHg. Exposure to media conditioned by lasered TMEs, or the addition of PGAs, induces the disassembly of intercellular junctions opening up the SCE barrier. Clinically, a positive PGA response predicts both a successful SLT outcome and the magnitude of the decrease in IOP after SLT. We hypothesize that SLT and PGA therapies may share a common mechanism of action.


Monoclonal antibodies specific for an acetylated form of alpha-tubulin recognize the antigen in cilia and flagella from a variety of organisms.
Seven monoclonal antibodies raised against tubulin from the axonemes of sea urchin sperm flagella recognize an acetylated form of alpha-tubulin present in the axoneme of a variety of organisms. The antigen was not detected among soluble, cytoplasmic alpha-tubulin isoforms from a variety of cells. The specificity of the antibodies was determined by in vitro acetylation of sea urchin and Chlamydomonas cytoplasmic tubulins in crude extracts. Of all the acetylated polypeptides in the extracts, only alpha-tubulin became antigenic. Among Chlamydomonas tubulin isoforms, the antibodies recognize only the axonemal alpha-tubulin isoform acetylated in vivo on the epsilon-amino group of lysine(s) (L'Hernault, S.W., and J.L. Rosenbaum, 1985, Biochemistry, 24:473-478). The antibodies do not recognize unmodified axonemal alpha-tubulin, unassembled alpha-tubulin present in a flagellar matrix-plus-membrane fraction, or soluble, cytoplasmic alpha-tubulin from Chlamydomonas cell bodies. The antigen was found in protein fractions that contained axonemal microtubules from a variety of sources, including cilia from sea urchin blastulae and Tetrahymena, sperm and testis from Drosophila, and human sperm. In contrast, the antigen was not detected in preparations of soluble, cytoplasmic tubulin, which would not have contained tubulin from stable microtubule arrays such as centrioles, from unfertilized sea urchin eggs, Drosophila embryos, and HeLa cells. Although the acetylated alpha-tubulin recognized by the antibodies is present in axonemes from a variety of sources and may be necessary for axoneme formation, it is not found exclusively in any one subset of morphologically distinct axonemal microtubules. The antigen was found in similar proportions in fractions from sea urchin sperm axonemes enriched for central pair or outer doublet B or outer doublet A microtubules. Therefore the acetylation of alpha-tubulin does not provide the mechanism that specifies the structure of any one class of axonemal microtubules. Preliminary evidence indicates that acetylated alpha-tubulin is not restricted to the axoneme. The antibodies described in this report may allow us to deduce the role of tubulin acetylation in the structure and function of microtubules in vivo.


Nitroxide stable radical suppresses autoimmune uveitis in rats.
Free radicals have been implicated in the pathogenesis of experimental autoimmune uveoretinitis (EAU). Nitroxides are stable radicals with a superoxide-dismutase-mimicking activity, which exert an anti-inflammatory effect in various animal models of oxidative damage and inflammation, such as experimental colitis and head trauma. We examined the use of the SOD mimic nitroxide 4-hydroxy-2,2,6,6,-tetramethylpiperidine-1-N-oxyl (TPL) to suppress EAU. Adult male Lewis rats were immunized with 125 microg/rat synthetic human retinal S-Ag, emulsified with Freund's adjuvant. Intravenous pertussis toxin was simultaneously injected. Beginning on Day 6, rats were injected with a daily intraperitoneal dose of 35, 175 or 350 micromol/rat of the nitroxide TPL. Control rats received intraperitoneal normal saline. The animals were examined daily, and on the 19th day the eyes were enucleated. Aqueous protein concentrations and retinal lipid peroxidation product levels (ketodienes and conjugated dienes) were determined. Histological sections were stained and examined microscopically. TPL was found to penetrate the aqueous humor readily. Beginning on day 12, rats developed a severe pan-uveitis. Rats in the treatment group had a lower mean clinical and histological score than that of controls. Levels of aqueous humor protein, retinal conjugated diens and ketodiens were all significantly lower in the treatment group. This effect was more pronounced with the lower TPL concentration. We conclude that TPL reduces clinical, biochemical and histopathological severity of S-Ag induced EAU in Lewis rats. This effect is probably mediated by removal of superoxide radicals, but other mechanisms may also be involved.


Congenital malformation and maternal occupational exposure to glycol ethers. Occupational Exposure and Congenital Malformations Working Group.
Glycol ethers are found in a wide range of domestic and industrial products, many of which are used in women's work environments. Motivated by concern about their potential reproductive toxicity, we have evaluated the risk of congenital malformations related to glycol ether exposure during pregnancy as part of a multicenter case-control study, conducted in six regions in Europe. The study comprised 984 cases of major congenital malformations and 1,134 controls matched for place and date of birth. Interviews of the mothers provided information about occupation during pregnancy, sociodemographic variables, and other potential risk factors (medical history, tobacco, alcohol, drugs). A chemist specializing in glycol ethers evaluated exposure during pregnancy, using the job description given by the mother, without knowledge of case or control status. We classified malformations into 22 subgroups. The overall odds ratio (OR) of congenital malformation associated with glycol ether exposure was 1.44 [95% confidence interval (CI) = 1.10-1.90], after adjustment for several potential confounders. The association with exposure to glycol ethers appeared particularly strong in three subgroups: neural tube defects (OR = 1.94; 95% CI = 1.16-3.24), multiple anomalies (OR = 2.00; 95% CI = 1.24-3.23), and cleft lip (OR = 2.03; 95% CI = 1.11-3.73). In this last subgroup, risk, especially of an isolated defect, tended to increase with level of exposure.


Chirality and chiroptical effects in inorganic nanocrystal systems with plasmon and exciton resonances.
This paper reviews the recent advances in experiment and theory of the induction of chiroptical effects, primarily circular dichroism (CD), at the plasmonic and excitonic resonances of achiral inorganic nanocrystals (NCs) capped and/or formed with chiral molecules. It also addresses stronger chiroptical effects obtained in intrinsically chiral inorganic nanostructures obtained from growing enantiomeric excess of intrinsically chiral NCs or arranging achiral plasmonic particles in chiral configurations. The accumulated experimental data and theory on various CD induction mechanisms provide an extended set of tools to properly analyze and understand the electromagnetic influence of chiral molecules on inorganic particles and obtain new general insights into the interaction of capping molecules with inorganic NCs. Among the field-induced CD mechanisms developed recently one can name the Coulomb (near-field, dipolar) mechanism for nanostructures much smaller than the wavelength, and for larger nanostructures, the electromagnetic (effective chiral medium), and intrinsically chiral plasmonic mechanisms.


Ibuprofen interferes with the efficacy of antihypertensive drugs. A randomized, double-blind, placebo-controlled trial of ibuprofen compared with acetaminophen.
To assess the effects of ibuprofen on blood pressure control in patients being treated with antihypertensive drugs. Randomized, blinded, placebo-controlled, parallel trial of ibuprofen compared with acetaminophen and with placebo in 3-week treatment periods. A general internal medicine clinic at a university hospital. Forty-five patients with essential hypertension controlled by treatment with at least two antihypertensive drugs were enrolled. Of these, 41 completed the study; treatment was discontinued in 3 of the 15 patients in the ibuprofen group due to breakage of the drug capsules, and after randomization in 1 of the 14 patients in the placebo group due to unstable angina. All 15 patients in the acetaminophen group completed the study. All previous antihypertensive regimens were continued. During the 3-week treatment, ibuprofen, 400 mg, was administered orally every 8 hours; acetaminophen, 1 g, orally every 8 hours; or placebo, 2 capsules, orally every 8 hours. In the ibuprofen group, the mean increase from baseline after 3 weeks of treatment was significant in the average supine diastolic blood pressure (6.4 mm Hg; 95% confidence interval [CI], 1.05 to 11.75; p = 0.0239); supine mean arterial pressure (6.6 mm Hg; 95% CI, 1.25 to 11.95; p = 0.0205); and sitting mean arterial pressure (5.8 mm Hg; 95% CI, 1.57 to 10.04; p = 0.0123). The mean increase in blood pressure variables in the ibuprofen group was significantly different compared with the mean increase in the variables in the placebo group after 3 weeks of treatment: supine systolic blood pressure (7.1 mm Hg compared with -4.5 mm Hg; 95% CI for the difference in means, 2.5 to 20.6; p = 0.0133); supine diastolic pressure (6.4 mm Hg compared with 0.0; 95% CI for difference in means, 0.87 to 12.4; p = 0.0250); supine mean arterial pressure (6.6 mm Hg compared with -1.5; 95% CI for difference in means, 2.0 to 14.2; p = 0.0110); sitting systolic pressure (6.8 mm Hg compared with -3.7; 95% CI for difference in means, 2.0 to 19.0; p = 0.0169); sitting diastolic pressure (5.3 mm Hg compared with -1.1; 95% CI for difference in means, 0.76 to 12.1; p = 0.0273); and sitting mean arterial pressure (5.8 mm Hg compared with -2.0; 95% CI for difference in means, 1.5 to 14.1; p = 0.0169).(ABSTRACT TRUNCATED AT 400 WORDS)


[Sociological analysis of healthcare delivery in Russia regions with inclement natural conditions (in case of the Republic of Altai)].
The paper describes the results of a sociological study of delivery of healthcare in the Republic of Altai to search for ways to improve its quality and to provide access to the local population. Analysis was made with regard to an interdisciplinary, comprehensive approach to considering the range of problems associated with not only the health of local residents and identifying risk factors leading to diseases, but also by determining the possibilities that could promote the minimization of causes that have a considerable impact on the occurrence of diseases and also hamper healthcare delivery in the human settlements of Gornyi Altai, which are difficult of access. The investigation has used sociological and statistical methods. It has been ascertained that the available healthcare forces and means should be today employed at the regional level, by applying the principles of necessary sufficiency in conjunction with the local population's social motivation to be involved in the activity associated with the rendering of medical services. In addition, it is necessary to systemically use mobile medical units as a significant factor for the optimization of medical care to the population living in Russia's regions which are difficult of access.


Structural evidence for an in trans base selection mechanism involving Loop1 in polymerase μ at an NHEJ double-strand break junction.
Eukaryotic DNA polymerase (Pol) X family members such as Pol μ and terminal deoxynucleotidyl transferase (TdT) are important components for the nonhomologous DNA end-joining (NHEJ) pathway. TdT participates in a specialized version of NHEJ, V(D)J recombination. It has primarily nontemplated polymerase activity but can take instructions across strands from the downstream dsDNA, and both activities are highly dependent on a structural element called Loop1. However, it is unclear whether Pol μ follows the same mechanism, because the structure of its Loop1 is disordered in available structures. Here, we used a chimeric TdT harboring Loop1 of Pol μ that recapitulated the functional properties of Pol μ in ligation experiments. We solved three crystal structures of this TdT chimera bound to several DNA substrates at 1.96-2.55 Å resolutions, including a full DNA double-strand break (DSB) synapsis. We then modeled the full Pol μ sequence in the context of one these complexes. The atomic structure of an NHEJ junction with a Pol X construct that mimics Pol μ in a reconstituted system explained the distinctive properties of Pol μ compared with TdT. The structure suggested a mechanism of base selection relying on Loop1 and taking instructions via the in trans templating base independently of the primer strand. We conclude that our atomic-level structural observations represent a paradigm shift for the mechanism of base selection in the Pol X family of DNA polymerases.


Effect of binocular cortical misalignment on ocular dominance and orientation selectivity.
We model a two-eye visual environment composed of natural images and study its effect on single cell synaptic modification. In particular, we study the effect of binocular cortical misalignment on receptive field formation after eye opening. We show that binocular misalignment affects principal component analysis (PCA) and Bienenstock, Cooper, and Munro (BCM) learning in different ways. For the BCM learning rule this misalignment is sufficient to produce varying degrees of ocular dominance, whereas for PCA learning binocular neurons emerge in every case.


A reproducing kernel Hilbert space framework for spike train signal processing.
This letter presents a general framework based on reproducing kernel Hilbert spaces (RKHS) to mathematically describe and manipulate spike trains. The main idea is the definition of inner products to allow spike train signal processing from basic principles while incorporating their statistical description as point processes. Moreover, because many inner products can be formulated, a particular definition can be crafted to best fit an application. These ideas are illustrated by the definition of a number of spike train inner products. To further elicit the advantages of the RKHS framework, a family of these inner products, the cross-intensity (CI) kernels, is analyzed in detail. This inner product family encapsulates the statistical description from the conditional intensity functions of spike trains. The problem of their estimation is also addressed. The simplest of the spike train kernels in this family provide an interesting perspective to others' work, as will be demonstrated in terms of spike train distance measures. Finally, as an application example, the RKHS framework is used to derive a clustering algorithm for spike trains from simple principles.


[An autopsy case of cryptococcal meningoencephalitis with AIDS: correspondence between MRI and pathological findings of basal ganglia and cerebellum].
We reported a 71-year-old male who showed subacute progression in cognitive decline and gait disturbance. Cystic lesions in the basal ganglia and a high signal in the right cerebellar hemisphere were detected respectively on MRI fluid-attenuated inversion recovery (FLAIR) image and diffusion weighted image (DWI) taken at 7 days after admission to our hospital. Dilatation of the Virchow-Robin space and fresh cerebellar infarction were suspected. Since an examination of cerebrospinal fluid (CSF) did not reveal any significant features, diagnosis of this patient proved very difficult. His cellular immunodeficiency was detected by bone marrow aspiration, so he was diagnosed as suffering from acquired immunodeficiency syndrome (AIDS). Six months after onset, the patient died from sepsis and respiratory failure. Pathological findings revealed multiple cryptococcus in the cystic lesions of the basal ganglia and cerebellum. There was little infiltration of inflammatory cells while pathological findings did not demonstrate any AIDS encephalopathy or vascular disorders. We speculated that subacute progression in cognitive decline and gait disturbance had been caused by cryptococcal meningoencephalitis and secondary normal pressure hydrocephalus (NPH). Cryptococcal meningoencephalitis in patients with AIDS is often accompanied by normal CSF as a result of cellular immunodeficiency, therefore diagnosis needs to be very careful. Particular attention needs to be paid to the existence of cryptococcal meningoencephalitis when patients show subacute progression in cognitive decline and cystic lesions in the basal ganglia present themselves on MRI.


Serum corticosteroid-binding globulin (CBG) and hepatic CBG mRNA relationships during hamster pregnancy: contribution of decidualization.
We measured serum corticosteroid-binding globulin (CBG) and hepatic CBG mRNA from individual hamsters throughout pregnancy and during decidualization. Serum CBG levels were determined by 3H-cortisol binding assay, and hepatic CBG mRNA levels were measured by Northern blots and solution hybridization assays, using a 32P-labeled cRNA probe derived from a rat CBG cDNA. There was a positive relationship between hepatic CBG mRNA levels and serum CBG levels during pregnancy. Both parameters increased significantly from the time of mating (cycle Day 4) to pregnancy Day 4, showing that CBG synthesis and secretion increased prior to implantation (Day 4). After implantation, serum CBG and hepatic CBG mRNA rose further from pregnancy Day 4 to a peak on Day 14, then decreased before parturition on Day 16. The prepartum decline in hepatic CBG mRNA preceded the fall in serum CBG. Decidualization on pseudopregnancy Day 4 resulted in an increase in serum CBG and hepatic CBG mRNA. Hepatic CBG mRNA increased from Day 5 to Day 7, and serum CBG increased progressively from Day 5 through Day 9 after uterine decidualization in the hamster. The present results demonstrate that the pattern of serum CBG observed in the pregnant hamster follows closely that of hepatic CBG mRNA. A signal emanating from uterine decidual tissue appears to be important in the regulation of hepatic CBG synthesis and secretion during midpregnancy, but other unknown factors appear to be involved in controlling CBG during the early and late stages of pregnancy.


Accurate mass-driven analysis for the characterization of protein phosphorylation. Study of the human Chk2 protein kinase.
We describe the data-dependent analysis of protein phosphorylation using rapid-acquisition nano-LC-linear quadrupole ion trap Fourier transform ion cyclotron resonance mass spectrometry (nano-LC-FTMS). The accurate m/z values of singly, doubly, and triply charged species calculated from the theoretical protonated masses of peptides phosphorylated at all Ser, Thr, or Tyr residues of the human checkpoint 2 (Chk2) protein kinase were used for selected ion extraction and chromatographic analysis. Using a kinase-inactive Chk2 mutant as a control, accurate mass measurements from FTMS and collision-induced dissociation spectra, 11 novel Chk2 autophosphorylation sites were assigned. Additionally, the presence of additional Chk2 phosphorylation sites in two unique peptides was deduced from accurate mass measurements. Selected ion chromatograms of all Chk2 phosphopeptides gave single peaks except in three cases in which two closely eluting species were observed. These pairs of phosphopeptides were determined to be positional isomers from MS/MS analysis. In this study, it was also found that ions due to the neutral loss of phosphoric acid from the parent peptide ion were not prominent in 18 of 36 MS/MS spectra of O-linked Chk2 phosphopeptides. Thus, accurate mass-driven analysis and rapid parallel MS/MS acquisition is a useful method for the discovery of new phosphorylation sites that is independent of the signature losses from phosphorylated amino acid residues.


Livestock welfare product claims: the emerging social context.
An increasing number of product claims about food animal welfare or well-being have appeared in the global food industry and global market in recent years. These claims have significant consequences for producers, processors, transporters, retailers, consumers, and the animals themselves. Furthermore, recent restructuring of the global food industry has altered the power relationships of various actors. Regulation of the industry is moving toward greater private control, and the power of retailers has dramatically increased. The changing structure of the industry carries implications both in terms of how standards are created and in terms of the types of standards themselves. The purpose of this article is to provide a greater understanding of how these product claims are made, their implications, and the challenges they present.


Borrelia-specific interferon-gamma and interleukin-4 secretion in cerebrospinal fluid and blood during Lyme borreliosis in humans: association with clinical outcome.
The Borrelia-specific interferon (IFN)- gamma and interleukin (IL)-4 responses of 113 patients and control subjects were analyzed using the sensitive enzyme-linked immunospot method. Cerebrospinal fluid (CSF) and blood samples were obtained, during the course of disease, from patients with chronic or nonchronic neuroborreliosis (NB) and from control subjects without NB. Blood samples were obtained from patients with Lyme skin manifestations and from healthy blood donors. Early increased secretion of Borrelia-specific IFN- gamma (P<.05) and subsequent up-regulation of IL-4 (P<.05) were detected in the CSF cells of patients with nonchronic NB. In contrast, persistent Borrelia-specific IFN- gamma responses were observed in the CSF cells of patients with chronic NB (P<.05). In patients with erythema migrans, increased IFN- gamma (P<.001) was observed in blood samples obtained early during the course of disease, whereas increased IL-4 (P<.05) was observed after clearance. On the contrary, patients with acrodermatitis chronica atrophicans had Borrelia-specific IFN- gamma (P<.001), but not IL-4, detected in blood samples. The present data suggest that an initial IFN- gamma response, followed by up-regulation of IL-4, is associated with nonchronic manifestations, whereas a persistent IFN- gamma response may lead to chronic Lyme borreliosis.


Thirteen-week oral dose toxicity study of Oligonol containing oligomerized polyphenols extracted from lychee and green tea.
Oligonol is a functional food containing catechin-type monomers and proanthocyanidin oligomer converted from polymer forms via a novel manufacturing process. The catechin component of green tea extract has been associated with nasal toxicity in rats following subchronic exposure. To assess the potential for Oligonol to induce nasal toxicity a 13-week repeated oral dose toxicity study was conducted in rats using doses of 100, 300, and 1000 mg/kg/d. Clinical signs and mortality were not affected by Oligonol treatment. Compound-colored stools and an increase in food consumption were observed in some treated groups; however, there were no treatment-related differences in terminal body weights or with respect to the results of the gross postmortem examinations. Histopathological evaluation of the nasal cavity tissues revealed no treatment-related lesions. The results from this toxicity study indicate that Oligonol does not induce nasal toxicity and further supports the results of previous studies demonstrating the safety of Oligonol for human consumption.


CD4-independent infection of astrocytes by human immunodeficiency virus type 1: requirement for the human mannose receptor.
Human immunodeficiency virus type 1 (HIV-1) infection occurs in the central nervous system and causes a variety of neurobehavioral and neuropathological disorders. Both microglia, the residential macrophages in the brain, and astrocytes are susceptible to HIV-1 infection. Unlike microglia that express and utilize CD4 and chemokine coreceptors CCR5 and CCR3 for HIV-1 infection, astrocytes fail to express CD4. Astrocytes express several chemokine coreceptors; however, the involvement of these receptors in astrocyte HIV-1 infection appears to be insignificant. In the present study using an expression cloning strategy, the cDNA for the human mannose receptor (hMR) was found to be essential for CD4-independent HIV-1 infectivity. Ectopic expression of functional hMR rendered U87.MG astrocytic cells susceptible to HIV-1 infection, whereas anti-hMR serum and hMR-specific siRNA blocked HIV-1 infection in human primary astrocytes. In agreement with these findings, hMR bound to HIV-1 virions via the abundant and highly mannosylated sugar moieties of HIV-1 envelope glycoprotein gp120 in a Ca(2+)-dependent fashion. Moreover, hMR-mediated HIV-1 infection was dependent upon endocytic trafficking as assessed by transmission electron microscopy, as well as inhibition of viral entry by endosomo- and lysosomotropic drugs. Taken together, these results demonstrate the direct involvement of hMR in HIV-1 infection of astrocytes and suggest that HIV-1 interaction with hMR plays an important role in HIV-1 neuropathogenesis.