Rapid conversion of autoimmune hypophysitis to an empty sella with immediate lowering of the serum IgG4 level. Case Report.
An 87-year-old man was admitted with fatigue, anorexia, vomiting, urinary incontinence, and a depressive state. His consciousness was evaluated as a 13 on the Glasgow Coma Scale (E4V3M6), and he had a body temperature of 36.4°C, a blood pressure of 91/60 mmHg, and a heart rate of 88 beats/min. General laboratory data were unremarkable except for a mildly elevated serum creatinine level. The plasma levels of growth hormone, luteinizing hormone, and follicle stimulating hormone were depressed. On the other hand, the prolactin level was elevated, and the corticotropin, cortisol, and thyrotropin levels were within the reference ranges. Cranial magnetic resonance imaging (MRI) revealed the marked swelling of the pituitary gland and the infundibular stalk, and the serum immunoglobulin G4 (IgG4) level was elevated (2.85 g/L; reference range, 0.048-1.05 g/L). Accordingly, a diagnosis of IgG4-related autoimmune hypophysitis (AH) was made. The patient responded well to glucocorticoid therapy, but the presence of diabetes insipidus was revealed and was subsequently controlled using desamino-D-arginine vasopressin (DDAVP). To our surprise, an empty sella was apparent on an MRI examination performed on Day 12. The patient's serum IgG4 level had decreased in a log-linear manner with a half-life of 30 days, which was comparable to the half-life of IgG4 in control subjects (21 days). At a 16-month follow-up examination, no substantial changes in the morphology or function of the pituitary gland were noted. In conclusion, an empty sella developed within 12 days after the clinical onset of AH in the present case, suggesting that an empty sella may be the direct outcome of AH. The conversion of AH to an empty sella was associated with an immediate shutdown of IgG4 overproduction.


Short telomeres in hatchling snakes: erythrocyte telomere dynamics and longevity in tropical pythons.
Telomere length (TL) has been found to be associated with life span in birds and humans. However, other studies have demonstrated that TL does not affect survival among old humans. Furthermore, replicative senescence has been shown to be induced by changes in the protected status of the telomeres rather than the loss of TL. In the present study we explore whether age- and sex-specific telomere dynamics affect life span in a long-lived snake, the water python (Liasis fuscus). Erythrocyte TL was measured using the Telo TAGGG TL Assay Kit (Roche). In contrast to other vertebrates, TL of hatchling pythons was significantly shorter than that of older snakes. However, during their first year of life hatchling TL increased substantially. While TL of older snakes decreased with age, we did not observe any correlation between TL and age in cross-sectional sampling. In older snakes, female TL was longer than that of males. When using recapture as a proxy for survival, our results do not support that longer telomeres resulted in an increased water python survival/longevity. In fish high telomerase activity has been observed in somatic cells exhibiting high proliferation rates. Hatchling pythons show similar high somatic cell proliferation rates. Thus, the increase in TL of this group may have been caused by increased telomerase activity. In older humans female TL is longer than that of males. This has been suggested to be caused by high estrogen levels that stimulate increased telomerase activity. Thus, high estrogen levels may also have caused the longer telomeres in female pythons. The lack of correlation between TL and age among old snakes and the fact that longer telomeres did not appear to affect python survival do not support that erythrocyte telomere dynamics has a major impact on water python longevity.


A de novo 11q23 deletion in a patient presenting with severe ophthalmologic findings, psychomotor retardation and facial dysmorphism.
Distal 11q deletion, previously known as Jacobsen syndrome, is caused by segmental aneusomy for the distal end of the long arm of chromosome 11. Typical clinical features include facial dysmorphism, mild-to-moderate psychomotor retardation, trigonocephaly, cardiac defects, and thrombocytopenia. There is a significant variability in the range of clinical features. We report herein a five-year-old girl with severe ophthalmological findings, facial dysmorphism, and psychomotor retardation with normal platelet function, in whom a de novo 11q23 deletion was detected, suggesting that distal 11q monosomy should be kept in mind in patients presenting with dysmorphic facial features and psychomotor retardation even in the absence of hematological findings.


Role of the stromal-derived factor-1 (SDF-1)-CXCR4 axis in the interaction between hepatic stellate cells and cholangiocarcinoma.
Cholangiocarcinoma (CCA) is highly fatal because of early invasion, widespread metastasis, and lack of an effective therapy. Migration, invasion, and metastasis of CCA cells are modulated by signals received from stromal cells. The SDF-1-CXCR4 axis emerges as a pivotal regulator of migration and survival of different tumor cells. The aim of the present study was to characterize the interaction between CCA cells and human hepatic stellate cells (hHSC) focusing on the role of SDF-1. The intrahepatic CCA cell line HuCCT-1 and primary hHSC were used for this study. RNA expression was examined by RTQ-PCR and protein expression by Western blotting. Immunofluorescence microscopy and immunohistochemistry were also employed. Migration of CCA cells was assessed using modified Boyden chambers. CXCR4 was clearly expressed in CCA cells of human CCA liver specimens. SDF-1 and hHSC conditioned medium (CM) promoted HuCCT-1 cell migration, which was abrogated by pre-incubation with AMD3100, a non-peptide antagonist of the CXCR4 receptor. In addition, HuCCT-1 cells silenced for CXCR4 did not migrate in presence of SDF-1. Both P-ERK and p-AKT were implicated in HuCCT-1 migration and showed a biphasic trend under stimulation of SDF-1. Finally, SDF-1 induced apoptotic rescue of HuCCT-1 cells by binding to CXCR4. Our study demonstrates that CCA cells migration and survival are modulated by the crosstalk between SDF-1, released by hHSC, and HuCCT-1 cells bearing CXCR4.


In vivo osteoinductive effect and in vitro isolation and cultivation bone marrow mesenchymal stem cells.
Bone marrow contains cell type termed mesenchymal stem cells (MSC), first recognized in bone marrow by a German pathologist, Julius Cohnheim in 1867. That MSCs have potential to differentiate in vitro in to the various cells lines as osteoblast, chondroblast, myoblast and adipoblast cells lines. Aims of our study were to show in vivo capacity of bone marrow MSC to produce bone in surgically created non critical size mandible defects New Zeland Rabbits, and then in second part of study to isolate in vitro MSC from bone marrow, as potential cell transplantation model in bone regeneration. In vivo study showed new bone detected on 3D CT reconstruction day 30, on all 3 animals non critical size defects, treated with bone marrow MSC exposed to the human Bone Morphogenetic Protein 7 (rhBMP-7). Average values of bone mineral density (BMD), was 530 mg/cm3, on MSC treated animals, and 553 mg/cm3 on control group of 3 animals where non critical size defects were treated with iliac crest autologue bone graft. Activity of the Alkaline Phosphatase enzyme were measurement on 0.5, 14, 21, 30 day and increased activity were detected day 14 on animals treated with bone marrow MSCs compared with day 30 on iliac crest treated animals. That results indicates strong osteoinduction activity of the experimental bone marrow MSCs models exposed to the rhBMP-7 factor Comparing ALP activity, that model showed superiorly results than control group. That result initiates us in opinion that MSCs alone should be alternative for the autolologue bone transplantation and in vitro study we isolated singles MSCs from the bone marrow of rat's tibia and femora and cultivated according to the method of Maniatopoulos et all. The small initial colonies of fibroblast like cells were photo-documented after 2 days of primary culture. Such isolated and cultivated MSCs in future studies will be exposed to the growth factors to differentiate in osteoblast and indicate their clinically potential as alternative for conventional medicine and autologue bone transplantation. That new horizons have potential to minimize surgery and patient donor morbidity, with more success treatment in bone regenerative and metabolism diseases.


Asymmetrical hip loading correlates with metal ion levels in patients with metal-on-metal hip resurfacing during sit-to-stand.
The occurrence of pseudotumours following metal-on-metal hip resurfacing arthroplasty (MoMHRA) has been associated with high serum metal ion levels and consequently higher than normal bearing wear. Measuring ground reaction force is a simple method of collecting information on joint loading during a sit-to-stand (STS). We investigated vertical ground reaction force (VGRF) asymmetry during sit-to-stand for 12 MoMHRA patients with known serum metal ion levels. Asymmetry was assessed using two methods: a ratio of VGRF for implanted/unimplanted side and an absolute symmetry index (ASI). It was found that subjects with high serum metal ion levels preferentially loaded their implanted sides. The difference between the two groups was most apparent during the first 22% of STS. VGRF ratio showed significant and strong correlation with serum metal ion levels (Spearman's rho = 0.8, p = 0.003). These results suggest that individual activity patterns play a role in the wear of MoMHRA and preferential loading of an implanted limb during the initiation of motion may increase the wear of metal-on-metal hip replacements.


Intramembrane changes in retinal pigment epithelial cell junctions of the dystrophic rat retina.
A progressive failure in phagocytosis by the retinal pigment epithelium (RPE) occurs in the Royal College of Surgeons rat with inherited retinal degeneration. Another change that can be attributed to a defect in the RPE is a breakdown in the blood-retinal barrier. RPE cell junctions, which form a part of this barrier, become permeable to extracellular tracer during the dystrophic process. We have used the freeze-fracture technique to study the structure of RPE cell junctions in normal and dystrophic retinas. In normal retinas, tight junctions between RPE cells consisted of 8 to 16 anastomosing strands on the cytoplasmic membrane leaflet (P-face) and a complementary pattern of grooves on the external membrane leaflet (E-face). Gap-junctional aggregates of hexagonally packed P-face particles and complementary E-face pits were enclosed within the tight junctional strands. In dystrophic retinas changes were first seen at postnatal day 21. Subtle breaks in P-face tight-junctional strands became more pronounced with time. Eventually the tight junctions appeared to unravel from the gap junctional aggregates, which became isolated and appeared to break off into patches of particle aggregates. The increased density of background particles in the membranes adjacent to disassembling junctions suggested that junctional elements were being removed by dispersal. Endocytosis of junctional elements was observed in both dystrophic and control retinas but may be accelerated in the dystrophic retina. In the late stages of the dystrophy some RPE cell junctions appeared to have proliferated and occupied extensive areas of the RPE membrane.


[The user's evaluation of the ophthalmologic service offered by SUS (Brazilian Unified Health System) in an urban center in the south of Brazil].
The user's evaluation is an important component of the quality of the health services. The goal of the present study was to evaluate the user's satisfaction with the ophthalmologic service provided by an accredited service of the Brazilian Unified Health System (SUS) in an urban center of the South of Brazil. It is a transversal study in which a questionnaire was applied about the user's socioeconomic and demographic profile, questions related to the quality of the service and suggestions and/or complaints regarding to the attendance and the service. The sample was calculated in the program EPINFO version 6.0, it had as its base 1200 monthly services, in a total of 355 users. The results showed that 77.1% of the users were satisfied with the medical service. However, 75.4% pointed deficiencies related to the time they had to wait in the place, to the duration of the consultation and at the interpersonal relationship with doctor and other attendants. It was verified that 36.3% considered the ophthalmologic service offered by SUS as terrible and regular. This way, it is fundamental to rethink the professional practices and to intervene on the form of organization of this service, seeking its improvement and the user's satisfaction.


Plaster of Paris-Short History of Casting and Injured Limb Immobilzation.
Various materials have been used since ancient times to help immobilise fractures. In this review, we discuss the history and developments of these materials as well as plaster of Paris. There has been a recent trend away from non-operative management of fractures, and skills in the use of plaster of Paris are declining. For the successful treatment of patients, it is important to appreciate how plaster works, how it should be used, and what can go wrong. In this review, we also discuss principles of applications and complications of plaster of Paris.


The HIV antibody test: why gay and bisexual men want or do not want to know their results.
Beginning in the latter part of 1985, 2,047 gay and bisexual men who were enrolled in the Pitt Men's Study, the Pittsburgh cohort of the Multicenter AIDS Cohort Study (MACS), were invited by mail to learn the results of their antibody test for HIV infection--human immunodeficiency virus infection. Participants were asked to complete and return a questionnaire designed to assess the factors influencing their (a) decision about learning the results, (b) recent sexual behavior, (c) knowledge about acquired immunodeficiency syndrome (AIDS), and (d) attitudes toward AIDS risk reduction. Of those men, 1,251 (61 percent) accepted the invitation, 188 (9 percent) declined, and 608 (30 percent) failed to respond. Fifty-four percent of the cohort subsequently learned their results. There were no significant differences in demographic, behavioral, and attitudinal characteristics or HIV seroprevalence between the men who accepted and those who declined. However, significant demographic differences were noted between the men who responded to the invitation versus those who did not; the latter group was composed of a greater proportion of men who were younger, nonwhite, and less educated. The most frequently cited reason (90 percent) why men wanted their test results was to determine if they had been infected with HIV. Of those who declined, 30 percent cited concerns about the psychological impact of learning about a positive result as being the most important factor for their decision. The two most frequently selected reasons for declining were the belief that the test is not predictive of the development of AIDS (48 percent) and concern about the worry that a positive result would produce (48 percent). These findings are discussed in the context of a nationwide, voluntary HIV screening program for gay and bisexual men.


Syntheses and Biological Activities of the LMNO, ent-LMNO, and NOPQR(S) Ring Systems of Maitotoxin.
Structure-activity relationship studies of maitotoxin (MTX), a marine natural product produced by an epiphytic dinoflagellate, were conducted using chemically synthesized model compounds corresponding to the partial structures of MTX. Both enantiomers of the LMNO ring system were synthesized via aldol reaction of the LM ring aldehyde and the NO ring ketone. These fragments were derived from a common cis-fused pyranopyran intermediate prepared through a sequence involving Nozaki-Hiyama-Kishi reaction, intramolecular oxa-Michael addition, and Pummerer rearrangement. The NOPQR(S) ring system, in which the original seven-membered S ring was substituted with a six-membered ring, was also synthesized through the coupling of the QR(S) ring alkyne and the NO ring aldehyde and the construction of the P ring via 1,4-reduction, dehydration, and hydroboration. The inhibitory activities of the synthetic specimens against MTX-induced Ca2+ influx were evaluated. The LMNO ring system and its enantiomer induced 36 and 18% inhibition, respectively, at 300 μM, whereas the NOPQR(S) ring system elicited no inhibitory activity.


Van Maldergem syndrome: further characterisation and evidence for neuronal migration abnormalities and autosomal recessive inheritance.
We present six patients from five unrelated families with a condition originally described by Van Maldergem et al and provide follow-up studies of the original patient. The phenotype comprises a distinctive facial appearance that includes blepharophimosis, maxillary hypoplasia, telecanthus, microtia and atresia of the external auditory meatus, intellectual disability, digital contractures and skeletal anomalies together with subependymal and subcortical neuronal heterotopia. Affected patients typically have neonatal hypotonia, chronic feeding difficulties and respiratory problems. In our cohort, we have observed one instance of sibling recurrence and parental consanguinity in three of the families, indicating that autosomal recessive inheritance is likely.


A Step-by-Step Procedure for Bone Regeneration Using Calcium Phosphate Scaffolds: From Site Preparation to Graft Placement.
Hydroxyapatite (HA) is the main mineral component of bone and teeth. HA is often used as a bone substitute and especially in its granular form it is osteoconductive and osseointegrating as shown by many investigations in animals and humans. HA granules are used for filling bone defects, but they have poor handling qualities and retention at the surgical site, leading to graft voids between the granules and bone tissue and resulting in mechanical instability. Furthermore HA implantation is not constantly followed by favorable results, especially when it is carried out for augmentation of the alveolar ridge. This article offers a protocol for bone reconstruction and predictable implant treatment outcomes. We provide a step-by-step description of both the recipient site and composite graft preparations using coralline HA granules (CHAG) and homologous fibrin glue (HFG). In the present study, 20 randomly selected patients (12 women and 8 men) underwent bone regeneration using CHAG-HFG before a dental implant procedure. Radiographic imaging, physical examination, and histological analysis were performed during a 2-year period. Biopsies were obtained at second-stage surgery before implant insertion using a 2.8-mm trephine bur. A morphological study of 20 bioptic human specimens was performed. Our results demonstrate that this surgical protocol for the preparation of the recipient site associated with a mixture of coralline HA granules with homologous fibrin glue provides reliable bone regeneration, thus reducing failures and minimizing risks of postoperative morbidity.


Hyperglycemia, recognized and unrecognized, as a risk factor for stroke and transient ischemic attacks.
Glycosylated hemoglobin concentration (GHb), which is considered an indication of glycemia over the preceding several months, was examined in 50 patients hospitalized for recent stroke or transient ischemic attacks (TIA), and compared to that in several reference populations. Patients with stroke or TIA had GHb (mean %A1 +/- SD, 10.2 +/- 2.3) higher than in hospital controls without cerebrovascular disease (8.3 +/- 0.9, p less than 0.005), and equivalent to values for ambulatory diabetic patients treated with diet or diet plus oral agents (9.5 +/- 2.4) or with insulin (10.7 +/- 2.9). Twenty percent (10/50) of the stroke/TIA group were previously known to have abnormal glucose tolerance or diabetes; when this subgroup was excluded, there remained 42% of the original group (21/50) with abnormal GHb (greater than 10% A1) not previously known to have hyperglycemia, and the difference between GHb values for the stroke/TIA patients not known to have glycemic abnormality and for the hospital control group remained significant (p less than 0.005). Sixty-two percent of stroke/TIA patients (31/50) were under treatment for glycemic abnormality, or had high GHb, or both. The high prevalence of elevated GHb in this population could not be attributed to a relationship to age, sex, smoking history, hypercholesterolemia, or hypertension. We conclude that hyperglycemia commonly precedes stroke and TIA, is usually unrecognized, and has been under-appreciated as a risk factor for cerebrovascular disease.


Deep down on a Caribbean reef: lower mesophotic depths harbor a specialized coral-endosymbiont community.
The composition, ecology and environmental conditions of mesophotic coral ecosystems near the lower limits of their bathymetric distributions remain poorly understood. Here we provide the first in-depth assessment of a lower mesophotic coral community (60-100 m) in the Southern Caribbean through visual submersible surveys, genotyping of coral host-endosymbiont assemblages, temperature monitoring and a growth experiment. The lower mesophotic zone harbored a specialized coral community consisting of predominantly Agaricia grahamae, Agaricia undata and a "deep-water" lineage of Madracis pharensis, with large colonies of these species observed close to their lower distribution limit of ~90 m depth. All three species associated with "deep-specialist" photosynthetic endosymbionts (Symbiodinium). Fragments of A. grahamae exhibited growth rates at 60 m similar to those observed for shallow Agaricia colonies (~2-3 cm yr(-1)), but showed bleaching and (partial) mortality when transplanted to 100 m. We propose that the strong reduction of temperature over depth (Δ5°C from 40 to 100 m depth) may play an important contributing role in determining lower depth limits of mesophotic coral communities in this region. Rather than a marginal extension of the reef slope, the lower mesophotic represents a specialized community, and as such warrants specific consideration from science and management.


Magnetic phase diagrams based on static and dynamic magnetic behaviour in Ru-based superconducting ferromagnets.
In this work, we present magnetic phase diagrams of a RuSr(2)Eu(1.5)Ce(0.5)Cu(2)O(10-δ) (Ru-1222) superconducting ferromagnet derived from its static and dynamic magnetic responses, measured by temperature and field dependences of dc magnetization and nonlinear ac susceptibility in both low and high magnetic fields. Comparison of magnetic phase diagrams of phase pure and impure samples singles out the intrinsic and extrinsic magnetic features, naturally proposing a unified model of Ru-1222 magnetic behaviour. The results considered within the proposed interpretation indicate full agreement between static and dynamic properties which, if measured in combination, effectively complement each other, uncovering existing ambiguities.


Reliability of mobile phone teledentistry in dental diagnosis and treatment planning in mixed dentition.
The aim of the study was to test the reliability of mobile phone teledentistry in the diagnosis and treatment planning of dental caries of children in mixed dentition. A total of 57 cases, aged 6-12 years, among six examiners were used yielding 342 comparisons. The patients were examined by a dentist who was calibrated in the recording of oral findings in children using the World Health Organization (WHO) oral health assessment form for children (version 2013), which served as the gold standard for diagnosis. Six paediatric dentists calibrated using the same WHO form served as the teledentistry group and made their diagnoses using only the images sent to them without the use of radiographs. The pictures obtained were stored on an online cloud platform (Google Drive). The sharing links for these pictures were forwarded to participating dentists using a social media application (WhatsApp Messenger, Facebook Corp., Mountain View, CA). This study showed greater sensitivity than specificity, and though both sensitivity and specificity were constantly above 80%, it can be stated that the current model has a higher chance of false positive results than false negative results. The reliability of teledentistry in this study is greater in primary teeth than in permanent teeth. Although the use of teledentistry without radiographs is not as accurate as clinical examination, mobile phone teledentistry offers acceptable reliability for the initial diagnosis of caries in children.


Chromosome assignment of the tumor-specific antigen of a 3-methylcholanthrene-induced mouse sarcoma.
Chemically induced sarcomas of inbred mice express antigens that are distinct and specific for each individual tumor. Chromosome assignment of tumor-specific antigens would help to elucidate the genetic basis of their diversity. Expression of the serologically defined Meth A sarcoma antigen is not suppressed in hybrid cells containing a complete foreign (Chinese hamster) genome. Chromosome and serologic analysis of microcell hybrids between Meth A sarcoma cells and Chinese hamster cells shows a clear correlation of Meth A antigen expression with the presence of the distal region of chromosome 12 (bands F1 and F2). Chromosome 16 may also be implicated. The significance of the close linkage of genes determining Meth A antigen expression with the immunoglobulin heavy chain gene cluster (on chromosome 12) and the lambda light chain genes (on chromosome 16) is discussed.


Detection of interictal epileptiform discharges using signal envelope distribution modelling: application to epileptic and non-epileptic intracranial recordings.
Interictal epileptiform discharges (spikes, IEDs) are electrographic markers of epileptic tissue and their quantification is utilized in planning of surgical resection. Visual analysis of long-term multi-channel intracranial recordings is extremely laborious and prone to bias. Development of new and reliable techniques of automatic spike detection represents a crucial step towards increasing the information yield of intracranial recordings and to improve surgical outcome. In this study, we designed a novel and robust detection algorithm that adaptively models statistical distributions of signal envelopes and enables discrimination of signals containing IEDs from signals with background activity. This detector demonstrates performance superior both to human readers and to an established detector. It is even capable of identifying low-amplitude IEDs which are often missed by experts and which may represent an important source of clinical information. Application of the detector to non-epileptic intracranial data from patients with intractable facial pain revealed the existence of sharp transients with waveforms reminiscent of interictal discharges that can represent biological sources of false positive detections. Identification of these transients enabled us to develop and propose secondary processing steps, which may exclude these transients, improving the detector's specificity and having important implications for future development of spike detectors in general.


Time from symptoms to definitive diagnosis of idiopathic pulmonary arterial hypertension: The delay study.
Survival rates for patients with idiopathic pulmonary arterial hypertension (IPAH) have improved with the introduction of PAH-specific therapies. However, the time between patient-reported onset of symptoms and a definitive diagnosis of IPAH is consistently delayed. We conducted a retrospective, multi-center, descriptive investigation in order to (a) understand what factors contribute to persistent diagnostic delays, and (b) examine the time from initial symptom onset to a definitive diagnosis of IPAH. Between January 2007 and December 2008, we enrolled consecutively diagnosed adults with IPAH from four tertiary referral centers in Australia. Screening of patient records and "one-on-one" interviews were used to determine the time from patient-described initial symptoms to a diagnosis of IPAH, confirmed by right heart catheterization (RHC). Thirty-two participants (69% female) were studied. Mean age at symptom onset was 56 ± 16.4 years and 96% reported exertional dyspnea. Mean time from symptom onset to diagnosis was 47 ± 34 months with patients subsequently aged 60 ± 17.3 years. Patients reported 5.3 ± 3.8 GP visits and 3.0 ± 2.1 specialist reviews before being seen at a pulmonary hypertension (PH) center. Advanced age, number of general practitioner (GP) visits, heart rate, and systolic blood pressure at the time of diagnosis were significantly associated with the observed delay. We found a significant delay of 3.9 years from symptom onset to a diagnosis of IPAH in Australia. Exertional dyspnea is the most common presenting symptom. Current practice within Australia does not appear to have the specific capacity for timely, multi-factorial evaluation of breathlessness and potential IPAH.


Longitudinal assessment of HIV-1 and HSV-2 shedding in the genital tract of West African women.
Forty-two HSV-2-seropositive women, of whom 22 were HIV-1 seropositive, were followed up weekly for 3 weeks for assessment of HIV-1 and HSV-2 genital shedding in Bobo Dioulasso, Burkina Faso. HSV-2 DNA and HIV-1 RNA were quantitated in cervicovaginal lavages enriched with a cervical swabbing (eCVL) using real-time polymerase chain reaction. HIV-1 RNA shedding was detected in eCVL from 19 of 22 (86.4%) of the HIV-1-seropositive women and HSV-2 DNA was detected in eCVL from 12 of 42 (28.6%) of the women. Compared with the HIV-1-seronegative women, the HIV-1-seropositive women showed more persistent HSV-2 genital shedding and higher HSV-2 DNA loads in eCVL. This study showed that eCVL is a reliable sampling method for detecting both viruses. Three measurement points (at weekly intervals in this study) seem to adequately allow detection of most HIV-1 or HSV-2 genital shedders.


Intracellular distribution of beta-actin mRNA is polarized in embryonic corneal epithelia.
The intracellular distribution of filamentous actin (F-actin), all actin isoforms and beta-actin mRNA were analyzed in whole-mount preparations of freshly isolated corneal epithelia. Filamentous actin distribution was analyzed with fluorescently tagged phalloidin. An antibody that recognizes an epitope on both globular (G-actin) and F-actin was used in an immunohistochemical analysis of actin protein distribution. Whole-mount epithelial tissues were examined with a confocal laser scanning microscope (CLSM). Biotinylated oligonucleotide probes specific for the beta-actin mRNA were used, and visualized with avidin-FITC. The intracellular localization of the beta-actin mRNA was similar to the F-actin protein distribution. In the most apical optical sections of embryonic cornea, actin staining delineated the cell borders and microvilli of the periderm cells. The actin is also detected as an organized network at the interface between the basal and periderm cells. At the level of the basal cell nucleus, F-actin is sparse, associating only with the lateral cell membranes. However, at the optical plane below the nuclei, the actin forms an elaborate actin cortical mat. Actin mRNA staining was visualized as discrete punctate areas. The beta-actin mRNA was positive at the optical plane just below the periderm cell apical membrane surface, similar to actin in microvilli. These cells also contained punctate staining near the cell membranes and in the periderm-basal cell junction area. At the level of the basal cell nucleus the actin mRNA was present in a punctate pattern along the cell membranes. Below the basal cell nuclei the actin mRNA staining increased at the level of the actin cortical mat. These experiments are the first demonstration that actin mRNA is polarized in embryonic corneal epithelia and co-localized with actin protein in an intact tissue.


Low-dose poly(ADP-ribose) polymerase inhibitor-containing combination therapies reverse early peripheral diabetic neuropathy.
Poly(ADP-ribose) polymerase (PARP) inhibition has recently been identified as a novel approach to treatment of experimental peripheral diabetic neuropathy (PDN). However, long-term inhibition of PARP, an enzyme involved in DNA repair, can potentially result in premature aging, loss of genome stability, and other side effects. This study evaluated potential synergistic interactions between low doses of the potent and specific PARP inhibitor 1,5-isoquinolinediol (ISO) and one of two vasodilators, the ACE inhibitor lisinopril (LIS) and the beta2-adrenoceptor agonist salbutamol (SAL) in the model of early PDN. Control and streptozotocin (STZ)-induced diabetic rats were treated with either ISO plus LIS or ISO plus SAL for 2 weeks after an initial 2 weeks without treatment. ISO (intraperitoneally) and LIS and SAL (both in the drinking water) were used in subtherapeutic doses, resulting in a minor correction of diabetes-associated sciatic motor and hind-limb digital sensory nerve conduction deficits when administered as monotherapies. Both combination treatments corrected endoneurial blood flow and vascular conductance deficits in STZ-induced diabetic rats. ISO plus SAL corrected all other changes of PDN, i.e., motor nerve conduction velocity (MNCV) and sensory nerve conduction velocity (SNCV) deficits as well as thermal and mechanical hyperalgesia. With ISO plus LIS, no significant correction of MNCV was observed, and the effect on thermal hyperalgesia was quite modest. SNCV and mechanical hyperalgesia were corrected. In vitro studies in human endothelial and Schwann cells showed early accumulation of poly(ADP-ribosyl)ated proteins (Western blot analysis) in response to high glucose, thus suggesting the importance of PARP activation in human PDN. In conclusion, low-dose PARP inhibitor-containing combination therapies may constitute a new approach for treatment of PDN.


Human brain dynamics accompanying use of egocentric and allocentric reference frames during navigation.
Maintaining spatial orientation while travelling requires integrating spatial information encountered from an egocentric viewpoint with accumulated information represented within egocentric and/or allocentric reference frames. Here, we report changes in high-density EEG activity during a virtual tunnel passage task in which subjects respond to a postnavigation homing challenge in distinctly different ways--either compatible with a continued experience of the virtual environment from a solely egocentric perspective or as if also maintaining their original entrance orientation, indicating use of a parallel allocentric reference frame. By spatially filtering the EEG data using independent component analysis, we found that these two equal subject subgroups exhibited differences in EEG power spectral modulation during tunnel passages in only a few cortical areas. During tunnel turns, stronger alpha blocking occurred only in or near right primary visual cortex of subjects whose homing responses were compatible with continued use of an egocentric reference frame. In contrast, approaching and during tunnel turns, subjects who responded in a way compatible with use of an allocentric reference frame exhibited stronger alpha blocking of occipito-temporal, bilateral inferior parietal, and retrosplenial cortical areas, all areas implicated by hemodynamic imaging and neuropsychological observation in construction and maintenance of an allocentric reference frame. We conclude that in these subjects, stronger activation of retrosplenial and related cortical areas during turns support a continuous translation of egocentrically experienced visual flow into an allocentric model of their virtual position and movement.


A complete series of uranium(iv) complexes with terminal hydrochalcogenido (EH) and chalcogenido (E) ligands E = O, S, Se, Te.
We here report the synthesis and characterization of a complete series of terminal hydrochalcogenido, U-EH, and chalcogenido uranium(iv) complexes, U≡E (with E = O, S, Se, Te), supported by the (Ad,MeArOH)3tacn (1,4,7-tris(3-(1-adamantyl)-5-methyl-2-hydroxybenzyl)-1,4,7-triazacyclononane) ligand system. Reaction of H2E with the trivalent precursor [((Ad,MeArO)3tacn)U] (1) yields the corresponding uranium(iv) hydrochalcogenido complexes [((Ad,MeArO)3tacn)U(EH)] (2). Subsequent deprotonation of the terminal hydrochalcogenido species with KN(SiMe3)2, in the presence of 2.2.2-cryptand, gives access to the uranium(iv) complexes with terminal chalcogenido ligands [K(2.2.2-crypt)][((Ad,MeArO)3tacn)U≡E] (3). In order to study the influence of the varying terminal chalogenido ligands on the overall molecular and electronic structure, all complexes were studied by single-crystal X-ray diffractometry, UV/vis/NIR, electronic absorption, and IR vibrational spectroscopy as well as SQUID magnetometry and computational analyses (DFT, MO, NBO).


Optimising the age, number of tests, and test interval for cervical screening in Canada.
Different approaches to screening for cancer of the cervix by cervical cytology have been evaluated using a computer simulation model developed by Knox and data on the natural history of carcinoma-in-situ (or worse) from a cohort study of women screened in British Columbia, 1949-69. The natural history input parameters and the output parameters without screening were modified to reflect the earlier onset of carcinoma-in-situ in younger cohorts now being experienced in British Columbia, resulting in simulated mortality from carcinoma of the cervix approximately 50% greater than that experienced in Canada in 1955. The simulations showed that the sensitivity of the test and the proportion of women in the population who accept invitations to attend for screening materially influence the extent to which programmes reduce mortality. Missed screens also have an important impact. With a 75% test sensitivity, and an 80% population acceptance, a programme designed to reduce mortality by 90% would commence at age 25, involve triennial screens to age 52, or triennial screens to age 40 and quinquennial screens to age 60, a total of 10 tests in a lifetime. A repeat test at age 26 contributes nothing to the mortality benefit. Nevertheless, additional modifications of the natural history specifications to accommodate high-risk younger women would require a more frequent schedule of examinations under the age of 35, though at a substantial 'cost' in terms of the total number of examinations required in a population.


QRS slopes for detection and characterization of myocardial ischemia.
In this study, the upward I(US) and downward I(DS) slopes of the QRS complex are proposed as indices for quantifying ischemia-induced electrocardiogram (ECG) changes. Using ECG recordings acquired before and during percutaneous transluminal coronary angioplasty (PTCA), it is found that the QRS slopes are considerably less steep during artery occlusion, in particular for I(DS). With respect to ischemia detection, the slope indices outperform the often used high-frequency index (defined as the root mean square (rms) of the bandpass-filtered QRS signal for the frequency band 150-250 Hz) as the mean relative factors of change are much larger for I(US) and I(DS) than for the high-frequency index (6.9 and 7.3 versus 3.7). The superior performance of the slope indices is equally valid when other frequency bands of the high-frequency index are investigated (the optimum one is found to be 125-175 Hz). Employing a simulation model in which the slopes of a template QRS are altered by different techniques, it is found that the slope changes observed during PTCA are mostly due to a widening of the QRS complex or a decrease of its amplitudes, but not a reduction of its high-frequency content or a combination of this and the previous effects. It is concluded that QRS slope information can be used as an adjunct to the conventional ST segment analysis in the monitoring of myocardial ischemia.


Up-regulation of small intestinal interleukin-17 immunity in untreated coeliac disease but not in potential coeliac disease or in type 1 diabetes.
Up-regulation of interleukin (IL)-17 in small intestinal mucosa has been reported in coeliac disease (CD) and in peripheral blood in type 1 diabetes (T1D). We explored mucosal IL-17 immunity in different stages of CD, including transglutaminase antibody (TGA)-positive children with potential CD, children with untreated and gluten-free diet-treated CD and in children with T1D. Immunohistochemistry was used for identification of IL-17 and forkhead box protein 3 (FoxP3)-positive cells and quantitative polymerase chain reaction (qPCR) for IL-17, FoxP3, retinoic acid-related orphan receptor (ROR)c and interferon (IFN)-γ transcripts. IL-1β, IL-6 and IL-17 were studied in supernatants from biopsy cultures. Expression of the apoptotic markers BAX and bcl-2 was evaluated in IL-17-stimulated CaCo-2 cells. The mucosal expression of IL-17 and FoxP3 transcripts were elevated in individuals with untreated CD when compared with the TGA-negative reference children, children with potential CD or gluten-free diet-treated children with CD (P < 0·005 for all IL-17 comparisons and P < 0·01 for all FoxP3 comparisons). The numbers of IL-17-positive cells were higher in lamina propria in children with CD than in children with T1D (P < 0·05). In biopsy specimens from patients with untreated CD, enhanced spontaneous secretion of IL-1β, IL-6 and IL-17 was seen. Activation of anti-apoptotic bcl-2 in IL-17-treated CaCo-2 epithelial cells suggests that IL-17 might be involved in mucosal protection. Up-regulation of IL-17 could, however, serve as a biomarker for the development of villous atrophy and active CD.


A simple core dataset and disease severity score for hereditary transthyretin (ATTRv) amyloidosis.
Hereditary transthyretin (ATTRv) amyloidosis is a progressive multisystemic disease of adult-onset that arises from an inherited mutation in the transthyretin gene. Currently available disease severity and progression evaluation tools only cover one single organ or system, impacting data collection uniformity and its use in clinical settings. The Jandhyala Method, including a systematic literature review and SMART interviews, was used to observe expert opinion from eight leaders in the treatment of ATTRv across Europe. The aim was to propose a multidisciplinary core dataset (CD) and disease severity scoring (DSS) tools. The multidisciplinary team of experts identified 140 indicators that form part of the standard diagnostic and monitoring practice (SDMP) and should be collected as the ATTRv CD. Thirty-one (22%) of these indicators informed disease severity and comprised the ATTRv DSS, whilst 25 (18%) were deemed to monitor disease progression. The resulting CD and DSS have different purposes. The ATTRv CD supports the collection of high-quality data for clinical research, whereas the ATTRv DSS can be rapidly conducted in a clinical setting and aid patient management.


Interrelationships between strength, anthropometrics, and strongman performance in novice strongman athletes.
The sport of strongman is relatively new; hence, specific research investigating this sport is currently very limited. The purpose of this study was to determine the relationships between anthropometric dimensions and maximal isoinertial strength to strongman performance in novice strongman athletes. Twenty-three semiprofessional rugby union players with considerable resistance training and some strongman training experience (age 22.0 ± 2.4 years, weight 102.6 ± 10.8 kg, height 184.6 ± 6.5 cm) were assessed for anthropometry (height, body composition, and girth measurements), maximal isoinertial performance (bench press, squat, deadlift, and power clean), and strongman performance (tire flip, log clean, and press, truck pull, and farmer's walk). The magnitudes of the relationships were determined using Pearson correlation coefficients, and interpreted qualitatively according to Hopkins (90% confidence limits ∼±0.37). The highest relationship observed was between system force (body mass + squat 1-repetition maximum) and overall strongman performance (r = 0.87). Clear moderate to very large relationships existed between performance in all strongman events and the squat (r = 0.61-0.85), indicating the importance of maximal squat strength for strongman competitors. Flexed arm girth and calf girth were the strongest anthropometric correlates of overall strongman performance (r = 0.79 and 0.70, respectively). The results of this study suggest that body structure and common gymnasium-based exercise strength are meaningfully related to strongman performance in novice strongman athletes. Future research should investigate these relationships using more experienced strongman athletes and determine the relationships between changes in anthropometry, isoinertial strength, and strongman performance to determine the role of anthropometry and isoinertial strength in the sport of strongman.


Mild phenotypes in a series of patients with Opitz GBBB syndrome with MID1 mutations.
Opitz syndrome (OS; MIM 145410 and MIM 300000) is a congenital midline malformation syndrome characterized by hypertelorism, hypospadias, cleft lip/palate, laryngotracheoesophageal (LTE) abnormalities, imperforate anus, developmental delay, and cardiac defects. The X-linked form (XLOS) is caused by mutations in the MID1 gene, which encodes a microtubule-associated RBCC protein. In this study, phenotypic manifestations of patients with and without MID1 mutations were compared to determine genotype-phenotype correlations. We detected 10 novel mutations, 5 in familial cases, 2 in sporadic cases, and 3 in families for whom it was not clear if they were familial or sporadic. The genotype and phenotype was compared for these 10 families, clinically diagnosed OS patients found not to have MID1 mutations, and 4 families in whom we have previously reported MID1 mutations. This combined data set includes clinical and mutation data on 70 patients. The XLOS patients with MID1 mutations were less severely affected than patients with MID1 mutations reported in previous studies, particularly in functionally significant neurologic, LTE, anal, and cardiac abnormalities. Minor anomalies were more prevalent in patients with MID1 mutations compared to those without mutations in this study. Female MID1 mutation carriers had milder phenotypes compared to male MID1 mutation carriers, with the most common manifestation being hypertelorism in both sexes. Most of the anomalies found in the patients of the present study do not correlate with the MID1 mutation type, with the possible exception of LTE malformations. This study demonstrates the wide spectrum of severity and manifestations of OS. It also shows that XLOS patients with MID1 mutations may be less severely affected than indicated in prior reports.


Large-sample-size assessment of socioeconomic predictors of community-level resource management occurrence.
Community-level resource management efforts are cornerstones in ensuring sustainable use of natural resources. Yet, understanding how community characteristics influence management practices remains contested. With a sample size of ≥725 communities, we assessed the effects of key community (i.e., socioeconomic) characteristics (human population size and density, market integration, and modernization) on the probability of occurrence of fisheries management practices, including gear, species, and spatial restrictions. The study was based in Solomon Islands, a Pacific Island country with a population that is highly dependent on coastal fisheries. People primarily dwell in small communities adjacent to the coastline dispersed across 6 island provinces and numerous smaller islands. We used nationally collected data in binomial logistic regression models to examine the likelihood of management occurrence, given socioeconomic context of communities. In contrast to prevailing views, we identified a positive and statistically significant association between both human population size and market integration and all 3 management practices. Human population density, however, had a statistically significant negative association and modernization a varied and limited association with occurrence of all management practices. Our method offers a way to remotely predict the occurrence of resource management practices based on key socioeconomic characteristics. It could be used to improve understanding of why some communities conduct natural resource management activities when statistical patterns suggest they are not likely to and thus improve understanding of how some communities of people beat the odds despite limited market access and high population density.


Fighting COVID-19 Misinformation on Social Media: Experimental Evidence for a Scalable Accuracy-Nudge Intervention.
Across two studies with more than 1,700 U.S. adults recruited online, we present evidence that people share false claims about COVID-19 partly because they simply fail to think sufficiently about whether or not the content is accurate when deciding what to share. In Study 1, participants were far worse at discerning between true and false content when deciding what they would share on social media relative to when they were asked directly about accuracy. Furthermore, greater cognitive reflection and science knowledge were associated with stronger discernment. In Study 2, we found that a simple accuracy reminder at the beginning of the study (i.e., judging the accuracy of a non-COVID-19-related headline) nearly tripled the level of truth discernment in participants' subsequent sharing intentions. Our results, which mirror those found previously for political fake news, suggest that nudging people to think about accuracy is a simple way to improve choices about what to share on social media.


Coordinate frame for pattern recognition in unilateral spatial neglect.
The present research examines the effect of spatial (object-centered) attentional constraints on pattern recognition. Four normal subjects and two right-hemisphere-damaged patients with left visual neglect participated in the study. Small, letterlike, prelearned patterns served as stimuli. Short exposure time prevented overt scanpaths during stimulus presentation. Attention was attracted to a central (midsagittal) hation point by precuing this location prior to each stimulus presentation. Minute (up to 1.5° of visual angle) rightward and leftward stimulus shifts caused attention to be allocated each time to a different location on the object space, while remaining in a fixed central position in viewercentered coordinates. The task was to decide which of several prelearned patterns was presented in each trial. In the normal subjects, best performance was achieved when the luminance centroid (LC; derived from the analysis of low-spatial frequencies in the object space) of each pattern coincided with the spatial position of the precue. In contrast, the patients with neglect showed optimal recognition performance when precuing attracted attention to locations within the object space, to the left of the LC. The normal performance suggests that the LC may serve as a center of gravity for attention allocation during pattern recognition. This point seems to be the target location where focal attention is normally directed, following a primary global analysis based on the low spatial frequencies. Thus, the LC of a simple pattern may serve as the origin point for an object-centered-coordiate-frame (OCCF), dividing it into right and left. This, in turn, serves to create a prototype description of the pattern, in its own coordinates, in memory, to be addressed during subsequent recognition tasks. The best match of the percept with the stored description may explain the observed advantage of allocating attention to the LC. The performance of the brain- damaged patients can be explained in terms of neglect operating in the OCCE.


Design of the PReferences for Open Versus Endovascular Repair of Abdominal Aortic Aneurysm (PROVE-AAA) Trial.
For patients with abdominal aortic aneurysm (AAA), randomized trials have found endovascular AAA repair (EVAR) is associated with lower perioperative morbidity and mortality than open surgical repair (OSR). However, OSR has fewer long-term aneurysm-related complications, such as endoleak or late rupture. Patients treated with EVAR and OSR have similar survival rates within two years after surgery, and OSR does not require intensive surveillance. Few have examined if patient preferences are aligned with the type of treatment they receive for their AAA. Although many assume that patients may universally prefer the less-invasive nature of EVAR, our preliminary work suggests that patients who value the lower risk of late complications may prefer OSR. In this study, called The PReferences for Open Versus Endovascular Repair of Abdominal Aortic Aneurysm (PROVE-AAA) trial, we describe a cluster-randomized trial to test if a decision aid can better align patients' preferences and their treatment type for AAA. Patients enrolled in the study are candidates for either endovascular or open repair and are followed up at VA hospitals by vascular surgery teams who regularly perform both types of repair. In Aim 1, we will determine patients' preferences for endovascular or open repair and identify domains associated with each repair type. In Aim 2, we will assess alignment between patients' preferences and the repair type elected and then compare the impact of a decision aid on this alignment between the intervention and control groups. This study will help us to accomplish two goals. First, we will better understand the factors that affect patient preference when choosing between EVAR and OSR. Second, we will better understand if a decision aid can help patients be more likely to receive the treatment strategy they prefer for their AAA. Study enrollment began on June 1, 2017. Between June 1, 2017 and November 1, 2018, we have enrolled 178 of a total goal of 240 veterans from 20 VA medical centers and their vascular surgery teams across the country. We anticipate completing enrollment in PROVE-AAA in June 2019, and study analyses will be performed thereafter.


Transcriptomic alterations in Trypanosoma cruzi-infected cardiac myocytes.
Trypanosoma cruzi infection is a major cause of cardiomyopathy. Previous gene profiling studies of infected mouse hearts have revealed prominent changes in gene expression within many functional pathways. This variety of transcriptomic changes in infected mice raises the question of whether gene expression alterations in whole hearts are due to changes in infected cardiac myocytes or other cells or even to systemic effects of the infection on the heart. We employed microarrays to examine infected cardiac myocyte cultures 48 h post-infection. Statistical comparison of gene expression levels of 7624 well annotated unigenes in four independent cultures of infected and uninfected myocytes detected substantial (>or=1.5 absolute fold changes) in 420 (5.5%) of the sampled genes. Major categories of affected genes included those involved in immune response, extracellular matrix and cell adhesion. These findings on infected cardiac myocytes in culture reveal that alterations in cardiac gene expression described in Chagas disease are the consequence of both direct infection of the myocytes themselves as well as resulting from the presence of other cell types in the myocardium and systemic effects of infection.


Optical fiber probe for biomedical Raman spectroscopy.
In vitro experiments have demonstrated the ability of Raman spectroscopy to diagnose a wide variety of diseases. Recent in vivo investigations performed with optical fiber probes were promising but generally limited to easily accessible organs, often requiring relatively long collection times. We have implemented an optical design strategy to utilize system throughput fully by characterizing the Raman distribution from tissue. This scheme optimizes collection efficiency, minimizes noise, and has resulted in small-diameter, highly efficient Raman probes that are capable of collecting high-quality data in 1 s. Performance has been tested through simulations and experiments with tissue models and several in vitro tissue types, demonstrating that this new design can advance Raman spectroscopy as a clinically practical technique.


Disparities in lung cancer staging with positron emission tomography in the Cancer Care Outcomes Research and Surveillance (CanCORS) study.
Disparities in treatment exist for nonwhite and Hispanic patients with non-small cell lung cancer, but little is known about disparities in the use of staging tests or their underlying causes. Prospective, observational cohort study of 3638 patients with newly diagnosed non-small cell lung cancer from 4 large, geographically defined regions, 5 integrated health care systems, and 13 VA health care facilities. Median age was 69 years, 62% were men, 26% were Hispanic or nonwhite, 68% graduated high school, 50% had private insurance, and 41% received care in the VA or another integrated health care system. After adjustment, positron emission tomography (PET) use was 13% lower among nonwhites and Hispanics than non-Hispanic whites (risk ratio [RR] 0.87, 95% confidence interval [CI] 0.77-0.97), 13% lower among those with Medicare than those with private insurance (RR 0.87, 95% CI 0.76-0.99), and 24% lower among those with an elementary school education than those with a graduate degree (RR 0.76, 95% CI 0.57-0.98). Disparate use of PET was not observed among patients who received care in an integrated health care setting, but the association between race/ethnicity and PET use was similar in magnitude across all other subgroups. Further analysis showed that income, education, insurance, and health care setting do not explain the association between race/ethnicity and PET use. Hispanics and nonwhites with non-small cell lung cancer are less likely to receive PET imaging. This finding is consistent across subgroups and not explained by differences in income, education, or insurance coverage.


Properties of a renin inhibitor isolated from the pig kidney cortex.
1. A renin-inhibitory material has been partially purified from soluble extracts of the pig kidney cortex by ammonium sulphate precipitation and diethylaminoethylcellulose (DEAE) chromatography and its properties studied. 2. It displayed competitive type kinetics. It did not inhibit cathepsin D, carboxypeptidase A, pancreatic kallikrein or trypsin. 3. Renins from dogs, rabbit and rat were inhibited, but not those from sheep or man, when assayed with pig angiotensinogen. 4. The material was inactivated by treatment with trypsin, N-ethylmaleimide or p-chloromercuribenzoate. 5. Renin-inhibitory activity was not found in plasma from peripheral blood of pigs. 6. It is concluded that the function of the renin inhibitor in the renal cortex of the pig may be restricted to the intrarenal environment.


Molecular cloning of the cDNA and chromosomal localization of the gene for a putative seven-transmembrane segment (7-TMS) receptor isolated from human spleen.
A family of proinflammatory cytokines sharing several structural features has been described and includes, for example, interleukin-8, monocyte chemoattractant protein-1, and melanocyte growth stimulatory activity. Recently, the receptors for interleukin-8 have been isolated and found to belong to the seven-transmembrane domain class of G protein-coupled receptors. As other members of this cytokine family likely interact with similar receptors, the polymerase chain reaction was employed to isolate related receptors from human peripheral blood adherent cells. Degenerate oligonucleotide primers based on the rabbit interleukin-8 receptor sequence were used. The corresponding full-length cDNA was isolated from a human spleen cDNA library. The predicted protein sequence of this clone, designated pBE1.3, was 93% identical to that of a cDNA isolated from bovine locus coeruleus, which apparently encodes a neuropeptide Y receptor, and also shows similarity with the interleukin-8 receptor and the human cytomegalovirus US28 sequences. The gene, designated D2S201E was localized to human chromosome 2q21. By Northern blotting, transcripts hybridizing to this cDNA were present in a variety of tissues and cells, including those of hemopoietic origin.


Structural diversity in a human antibody germline library.
To support antibody therapeutic development, the crystal structures of a set of 16 germline variants composed of 4 different kappa light chains paired with 4 different heavy chains have been determined. All four heavy chains of the antigen-binding fragments (Fabs) have the same complementarity-determining region (CDR) H3 that was reported in an earlier Fab structure. The structure analyses include comparisons of the overall structures, canonical structures of the CDRs and the VH:VL packing interactions. The CDR conformations for the most part are tightly clustered, especially for the ones with shorter lengths. The longer CDRs with tandem glycines or serines have more conformational diversity than the others. CDR H3, despite having the same amino acid sequence, exhibits the largest conformational diversity. About half of the structures have CDR H3 conformations similar to that of the parent; the others diverge significantly. One conclusion is that the CDR H3 conformations are influenced by both their amino acid sequence and their structural environment determined by the heavy and light chain pairing. The stem regions of 14 of the variant pairs are in the 'kinked' conformation, and only 2 are in the extended conformation. The packing of the VH and VL domains is consistent with our knowledge of antibody structure, and the tilt angles between these domains cover a range of 11 degrees. Two of 16 structures showed particularly large variations in the tilt angles when compared with the other pairings. The structures and their analyses provide a rich foundation for future antibody modeling and engineering efforts.


HIV risk and associations of HIV infection among men who have sex with men in peri-urban Cape Town, South Africa.
The HIV epidemic in Sub Saharan Africa has been traditionally assumed to be driven by high risk heterosexual and vertical transmission. However, there is an increasing body of data highlighting the disproportionate burden of HIV infection among MSM in the generalized HIV epidemics across of Southern Africa. In South Africa specifically, there has been an increase in attention focused on the risk status and preventive needs of MSM both in urban centers and peri-urban townships. The study presented here represents the first evaluation of HIV prevalence and associations of HIV infection among MSM in the peri-urban townships of Cape Town. The study consisted of an anonymous probe of 200 men, reporting ever having had sex with another man, recruited through venue-base sampling from January to February, 2009. Overall, HIV prevalence was 25.5% (n = 51/200). Of these prevalent HIV infections, only 6% of HIV-1 infected MSM were aware of their HIV status (3/50). 0% of men reported always having safe sex as defined by always wearing condoms during sex and using water-based lubricants. Independent associations with HIV infection included inconsistent condom use with male partners (aOR 2.3, 95% CI 1.0-5.4), having been blackmailed (aOR 4.4, 95% CI 1.6-20.2), age over 26 years (aOR 4.2, 95% CI 1.6-10.6), being unemployed (aOR 3.7, 95% CI 1.5-9.3), and rural origin (aOR 6.0, 95% CI 2.2-16.7). Bisexual activity was reported by 17.1% (34/199), and a total of 8% (16/200) reported having a regular female partner. Human rights violations were common with 10.5% (n = 21/200) reporting having been blackmailed and 21.0% (n = 42/200) reporting being afraid to seek health care. The conclusions from this study include that a there is a high risk and underserved population of MSM in the townships surrounding Cape Town. The high HIV prevalence and high risk sexual practices suggest that prevalence will continue to increase among these men in the context of an otherwise slowing epidemic. These data further highlight the need to better characterize risk factors for HIV prevention and appropriate targeted combination packages of HIV interventions including biomedical, behavioural, and structural approaches to mitigate HIV risk among these men.


Food web structure and interaction strength pave the way for vulnerability to extinction.
This paper focuses on how food web structure and interactions among species affects the vulnerability, due to environmental variability, to extinction of species at different positions in model food webs. Vulnerability is here not measured by a traditional extinction threshold but is instead inspired by the IUCN criteria for endangered species: an observed rapid decline in population abundance. Using model webs influenced by stochasticity with zero autocorrelation, we investigate the ecological determinants of species vulnerability, i.e. the trophic interactions between species and food web structure and how these interact with the risk of sudden drops in abundance of species. We find that (i) producers fulfil the criterion of vulnerable species more frequently than other species, (ii) food web structure is related to vulnerability, and (iii) the vulnerability of species is greater when involved in a strong trophic interaction than when not. We note that our result on the relationship between extinction risk and trophic position of species contradict previous suggestions and argue that the main reason for the discrepancy probably is due to the fact that we study the vulnerability to environmental stochasticity and not extinction risk due to overexploitation, habitat destruction or interactions with introduced species. Thus, we suggest that the vulnerability of species to environmental stochasticity may be differently related to trophic position than the vulnerability of species to other factors. Earlier research on species extinctions has looked for intrinsic traits of species that correlate with increased vulnerability to extinction. However, to fully understand the extinction process we must also consider that species interactions may affect vulnerability and that not all extinctions are the result of long, gradual reductions in species abundances. Under environmental stochasticity (which importance frequently is assumed to increase as a result of climate change) and direct and indirect interactions with other species some extinctions may occur rapidly and apparently unexpectedly. To identify the first declines of population abundances that may escalate and lead to extinctions as early as possible, we need to recognize which species are at greatest risk of entering such dangerous routes and under what circumstances. This new perspective may contribute to our understanding of the processes leading to extinction of populations and eventually species. This is especially urgent in the light of the current biodiversity crisis where a large fraction of the world's biodiversity is threatened.


Use of optical scattering to discriminate particle types in coastal waters.
The particulate scattering characteristics of coastal waters were examined at nine locations around the United States, including near-shore sites in the Gulf of Mexico and the Atlantic and Pacific oceans. The scattering measurements were used in conjunction with inversion models to estimate particle size distributions and bulk refractive indices of the suspended particles. The relationships between various scattering properties and chlorophyll concentration were also investigated and compared with previous relationships described for case I waters. Although the general patterns of scattering and particle characteristics in coastal waters were fairly consistent, fine-scale variability within the water column was substantial. Combining optical measurements with inversion techniques provided a more informative view of the environment and a better understanding of the nature of particle populations in the coastal ocean.


Health related quality of life in Middle Eastern children with beta-thalassemia.
Thalassemia is a common disorder worldwide with a predominant incidence in Mediterranean countries, North Africa, the Middle East, India, Central Asia, and Southeast Asia. Whilst substantial progress has been made towards the improvement of Health related quality of life (HRQoL) in western countries, scarce evidence-based data exists on HRQol of thalassemia children and adolescents living in developing countries. We studied 60 thalassemia children from Middle Eastern countries with a median age of 10 years (range 5 to 17 years). HRQoL was assessed with the Pediatric Quality of Life Inventory (PedsQL) 4.0. The Questionnaire was completed at baseline by all patients and their parents. The agreement between child-self and parent-proxy HRQoL reports and the relationship between HRQoL profiles and socio-demographic and clinical factors were investigated. The scores of parents were generally lower than those of their children for Emotional Functioning (mean 75 vs 85; p = 0.002), Psychosocial Health Summary (mean 70.3 vs 79.1; p = 0.015) and the Total Summary Score (mean 74.3 vs 77.7 p = 0.047). HRQoL was not associated with ferritin levels, hepatomegaly or frequency of transfusions or iron chelation therapy. Multivariate analysis showed that a delayed start of iron chelation had a negative impact on total PedsQL scores of both children (p = 0.046) and their parents (p = 0.007). The PedsQL 4.0 is a useful tool for the measurement of HRQoL in pediatric thalassemia patients. This study shows that delayed start of iron chelation has a negative impact on children's HRQoL.


The histone demethylase Jmjd3 sequentially associates with the transcription factors Tbx3 and Eomes to drive endoderm differentiation.
Stem cell differentiation depends on transcriptional activation driven by lineage-specific regulators as well as changes in chromatin organization. However, the coordination of these events is poorly understood. Here, we show that T-box proteins team up with chromatin modifying enzymes to drive the expression of the key lineage regulator, Eomes during endodermal differentiation of embryonic stem (ES) cells. The Eomes locus is maintained in a transcriptionally poised configuration in ES cells. During early differentiation steps, the ES cell factor Tbx3 associates with the histone demethylase Jmjd3 at the enhancer element of the Eomes locus to allow enhancer-promoter interactions. This spatial reorganization of the chromatin primes the cells to respond to Activin signalling, which promotes the binding of Jmjd3 and Eomes to its own bivalent promoter region to further stimulate Eomes expression in a positive feedback loop. In addition, Eomes activates a transcriptional network of core regulators of endodermal differentiation. Our results demonstrate that Jmjd3 sequentially associates with two T-box factors, Tbx3 and Eomes to drive stem cell differentiation towards the definitive endoderm lineage.


[Evaluation of performance of health systems: a model for analysis].
This paper presents a review of the Dimension Matrix for Evaluation of the Brazilian Health System that was initially developed in 2003, as well as a conceptual update of some of the sub-dimensions for the evaluation of health service performance, namely effectiveness, access, efficiency and appropriateness of health care. It also describes the indicator selection process as well as the results obtained in each performance dimension. The behavior of the indicators used to assess the performance of health services in Brazil, with respect to each sub-dimension, was not uniform. Areas of marked improvement were found in indicators that are influenced by activities in the field of primary care. The most significant improvements were seen in the sub-dimensions of Effectiveness and Access. With respect to the Efficiency of health services, situations of high efficiency coexist with others with substandard performance. The performance of health services in the sub-dimension of Appropriateness of Health Care was the lowest of all indicators.


Cryptic female preference for genetically unrelated males is mediated by ovarian fluid in the guppy.
As inbreeding is costly, it has been suggested that polyandry may evolve as a means to reduce the negative fitness consequences of mating with genetically related males. While several studies provide support for this hypothesis, evidence of pure post-copulatory mechanisms capable of biasing paternity towards genetically unrelated males is still lacking; yet these are necessary to support inbreeding avoidance models of polyandry evolution. Here we showed, by artificially inseminating a group of female guppies with an equal number of sperm from related (full-sib) and unrelated males, that sperm competition success of the former was 10 per cent lower, on average, than that of the unrelated male. The paternity bias towards unrelated males was not due to differential embryo survival, as the size of the brood produced by control females, which were artificially inseminated with the sperm of a single male, was not influenced by their relatedness with the male. Finally, we collected ovarian fluid (OF) from virgin females. Using computer-assisted sperm analysis, we found that sperm velocity, a predictor of sperm competition success in the guppy, was significantly lower when measured in a solution containing the OF from a sister as compared with that from an unrelated female. Our results suggest that sperm-OF interaction mediates sperm competition bias towards unrelated mates and highlight the role of post-copulatory mechanisms in reducing the cost of mating with relatives in polyandrous females.


The nature of shape constancy mechanisms as revealed by shape priming.
Five shape priming experiments are reported in which the target was either a five- or six-sided line-drawn figure and participants made a speeded two-alternative forced-choice judgment about the target's number of sides. On priming trials, the target was preceded by a briefly presented smaller line figure (the prime) and performance on these trials was gauged relative to a no-prime condition. In the first two experiments, primes were rendered invisible by the presentation of a backwards visual noise mask, respectively for a short (∼40 ms) or long duration (∼93 ms). No reliable priming effects arose under masked conditions. When these experiments were repeated without the mask, participants were speeded when the prime and target were related by a rigid through-the-plane rotation but not when the prime was a nonrigid, stretched version of the target. The same pattern of priming effects arose when, in a final experiment, novel irregular shapes were used. Collectively, the data reveal the operation of shape constancy mechanisms that are particularly sensitive to shape rigidity. The findings suggest that the visual system attempts to secure a correspondence between the rapid and successive presentations of the prime and the target by matching shapes according to a rigidity constraint.


A deletion linked to a poly(ADP-ribose) polymerase gene on chromosome 13q33-qter occurs frequently in the normal black population as well as in multiple tumor DNA.
The nuclear enzyme poly(ADP-ribose) polymerase (PADPRP) is thought to play a role in DNA recombination, replication, and repair. In view of the implication of these processes in tumorigenesis, and based on preliminary evidence which indicated the presence of an extraneous polymorphic restriction fragment for murine PADPRP loci in strains of mice susceptible to plasmacytomas, we investigated correlations between the restriction fragment length polymorphism of the PADPRP gene(s) and human Burkitt lymphoma. No increase in the frequency of polymorphisms on chromosome 1 (containing the active gene) or on chromosome 14 (a pseudogene) was observed. However, restriction fragment length polymorphism analysis of PADPRP sequences on chromosome 13 (either a processed pseudogene or a gene with extensive identity to PADPRP) revealed that of 19 DNA samples derived from endemic Burkitt lymphoma all contained at least one copy of a rare allele (B). Simple two-allele (A/B) polymorphisms in this PADPRP-like locus were identified by digestion with a number of restriction enzymes including HindIII, PstI, KpnI, and MspI. These restriction fragment length polymorphisms always segregated together, suggesting that they identify a deletion within or close to the PADPRP sequences on chromosome 13, which we mapped precisely to 13q33-qter. Based upon family studies the A and B alleles were shown to be transferred in a Mendelian codominant fashion. Subsequently, this probe was used as a linkage marker to study the frequency of this deletion in various tumors including B-cell follicular lymphomas, small cell lung carcinomas, breast carcinomas, and colorectal carcinomas. In noncancer control populations, the frequency of this deletion was 3-fold higher among Blacks as compared to Caucasians. When DNA from various tumors was compared to normal DNA from racially appropriate noncancer controls, the frequency of this deletion was still 2- to 3-fold higher in the tumor DNA. Matched samples provided instances of tumor-specific loss of heterozygosity but also revealed that the predominant source of this deletion is the germ line, suggesting that the chromosome 13 region neighboring the PADPRP locus may harbor a gene whose loss may predispose individuals to malignancy.


An ERP study of the interaction between verbal information and conditioning pathways to fear.
Two experiments are described that explore the effects of verbal information and direct conditioning in the acquisition and extinction of fear responses. Participants were given verbal threat information about novel animals before conditioning trials in which the animals were presented alongside an aversive outcome (Experiment 1), or positive information about the animals before extinction trials (Experiment 2). Fear was measured using self-reported fear beliefs, expectancy of the unconditioned stimulus (US) and event-related brain potential (ERP). The results showed a direct effect of verbal information on acquisition (Experiment 1) and extinction (Experiment 2). There was a P2 peak latency shift at acquisition (Experiment 1) and P1 mean amplitude response at extinction (Experiment 2) based on the interaction between verbal information and US-contingency. However, the P2 response showed little evidence for an enhanced conditioned response (CR) when verbal threat information and direct conditioning combined: earlier P2 responses were found for all animals that had been associated with either threat information or the aversive US. Additionally, increase in P1 mean amplitude response (Experiment 2) seemed to stem from the conflict between verbal information and contingency information, rather than the predicted decrease in response where positive information and extinction training were combined. Future studies are suggested that might explore whether attention/arousal modulate the P1 response as a result of such expectation violations.


Temporal profile of amygdala γ oscillations in response to faces.
Neuroimaging studies have reported greater activation of the human amygdala in response to faces than to nonfacial stimuli, yet little is known about the temporal profile of this activation. We investigated this issue by recording the intracranial field potentials of the amygdala in participants undergoing preneurosurgical assessment (n = 6). Participants observed faces, mosaics, and houses in upright and inverted orientations using a dummy target detection task. Time-frequency statistical parametric mapping analyses revealed that the amygdala showed greater gamma-band activity in response to faces than to mosaics at 200-300 msec, with a peak at 255 msec. Gamma-band activation with a similar temporal profile was also found in response to faces versus houses. Activation patterns did not differ between upright and inverted presentations of stimuli. These results suggest that the human amygdala is involved in the early stages of face processing, including the modulation of subjective perception of faces.


Isolation and phylogeny of new endogenous retroviral sequences belonging to the HERV-F family.
A new human endogenous retroviral family (HERV-F) has been identified from human chromosome 7q31.1-q31.3 that was identical to the XA34 cDNA clone isolated from a human glioma cDNA library with an ERV-9 env probe. We investigated pol gene sequences of the HERV-F family from a human monochromosomal DNA panel and analyzed these with HERV-F. The pol gene sequences of the HERV-F family were detected on chromosomes 3, 6, 7, 10, 11, 14, 19, 20, X, and Y as examined by PCR. Thirty-six pol gene sequences identified from the human chromosomes have a high degree of sequence similarity (80-99%) with that of the HERV-F. Phylogenetic analysis of pol gene sequences distinctively showed four groups, indicating that the HERV-F family could be amplified at least four times after the original integration into the human genome or represent integration events separately during hominid evolution. One clone (HFY-3) on chromosome Y shared 100% sequence identity with a clone (HF19-2) on chromosome 19, and a clone (HF20-6) on chromosome 20 suggests either a recent retrotransposition or a chromosomal translocation. The history of endogenous retroviral sequences may contribute to an understanding of evolutionary change in human genomes.


Methodology Perspective of Computing Thermal Transport in Low-Dimensional Materials and Nanostructures: The Old and the New.
Demands for engineering thermal transport properties are ever increasing for a wide range of modern micro- and nanodevices and materials-based energy technologies. In particular, there is a severe situation due to the rapid progress in the synthesis and processing of materials and devices with structural characteristic length on the nanometer scales, which are comparable or even smaller than the intrinsic length scales (such as mean free path and wavelength) of basic energy carriers (such as phonons, electrons, and photons). Although advanced approaches for controlling the electronic and photonic transport have been proposed in the past decades, progress on controlling lattice vibrations (i.e., the phonons) is still far behind. Gaps between the fundamental understandings of the behavior of the basic energy carriers at small scales and the technological demands still remain, particularly from a computer modeling point of view. Herewith, we give a perspective of the computational approaches for predicting the thermal transport properties of low-dimensional materials and nanostructures, which are mainly sorted into three categories: empirical molecular dynamics, anharmonic lattice dynamics based Boltzmann transport equation, and Landauer theory. The advantage and disadvantage of each method are discussed and some possible solutions are suggested. The discussion is focused on fully and accurately characterizing the mode-level phonon behavior, possible all-order phonon scattering process, and incorporation of realistic nanostructures. Moreover, emerging challenges of phonon coupling effects, such as electron-phonon, phonon-photon, and phonon-magnon coupling, are also discussed. We expect that this perspective will stimulate future research in computer modeling of micro-/nanoscale heat transfer beyond traditional phonons.


Perspectives on the methods of a large systematic mapping of maternal health interventions.
Mapping studies describe a broad body of literature, and differ from classical systematic reviews, which assess more narrowly-defined questions and evaluate the quality of the studies included in the review. While the steps involved in mapping studies have been described previously, a detailed qualitative account of the methodology could inform the design of future mapping studies. Describe the perspectives of a large research team on the methods used and collaborative experiences in a study that mapped the literature published on maternal health interventions in low- and middle-income countries (2292 full text articles included, after screening 35,048 titles and abstracts in duplicate). Fifteen members of the mapping team, drawn from eight countries, provided their experiences and perspectives of the study in response to a list of questions and probes. The responses were collated and analysed thematically following a grounded theory approach. The objectives of the mapping evolved over time, posing difficulties in ensuring a uniform understanding of the purpose of the mapping among the team members. Ambiguity of some study variables and modifications in data extraction codes were the main threats to the quality of data extraction. The desire for obtaining detailed information on a few topics needed to be weighed against the benefits of collecting more superficial data on a wider range of topics. Team members acquired skills in systematic review methodology and software, and a broad knowledge of maternal health literature. Participation in analysis and dissemination was lower than during the screening of articles for eligibility and data coding. Though all respondents believed the workload involved was high, study outputs were viewed as novel and important contributions to evidence. Overall, most believed there was a favourable balance between the amount of work done and the project's outputs. A large mapping of literature is feasible with a committed team aiming to build their research capacity, and with a limited, simplified set of data extraction codes. In the team's view, the balance between the time spent on the review, and the outputs and skills acquired was favourable. Assessments of the value of a mapping need, however, to take into account the limitations inherent in such exercises, especially the exclusion of grey literature and of assessments of the quality of the studies identified.


Fabrication and detection of tissue-engineered bones with bio-derived scaffolds in a rotating bioreactor.
In order to explore the methods for commercialized bone tissue engineering, engineered bones should be cultivated in bioreactors to realize three-dimensional culture under well-defined culture conditions. In the present paper, osteoblasts isolated from the cranium of 1-month-old Zelanian rabbits were inoculated on to the BDBS (bio-derived bone scaffolds) to investigate the three-dimensional fabrication of engineered bone in an RWVB (rotating-wall vessel bioreactor). The osteoblasts, after being transfected with green fluorescent protein, were respectively seeded at 2 x 10(6) and 1 x 10(6) cells x ml(-1) on to the BDBS and then cultured in a T-flask and an RWVB for 1 week. The morphologies and structure of the fabricated bone were investigated by using an inverted microscope, a scanning electron microscope and a laser confocal microscope using the stains haematoxylin/eosin and Toluidine Blue. After being digested from the scaffolds, the cells were assayed with ALP (alkaline phosphatase) stain, von-Kossa staining on mineralized nodules, type I collagen and bone morphogenetic protein-2 expression, and the cell expansion and growth curves using different culture methods were quantitatively determined with MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide). Furthermore, cell cycle and apoptosis were detected by using a flow cytometer, and total DNA was also assayed. For a comparative study, cell-seeded constructs were also cultured under static conditions. The results show that the cell number cultured in the RWVB was five times that in the T-flask. Bone tissues cultured in the RWVB with two different densities grew well, and the osteoblasts maintained their normal cycle and DNA content. The result demonstrates that, with the stress stimulation in the fluid in the RWVB, the active expression of ALP can be increased, rapid proliferation and differentiation of osteoblasts are possible and the three-dimensional fabrication of engineered bone could be realized.


Estimating the Fitness Effect of Deleterious Mutations During the Two Phases of the Life Cycle: A New Method Applied to the Root-Rot Fungus Heterobasidion parviporum.
Many eukaryote species, including taxa such as fungi or algae, have a lifecycle with substantial haploid and diploid phases. A recent theoretical model predicts that such haploid-diploid lifecycles are stable over long evolutionary time scales when segregating deleterious mutations have stronger effects in homozygous diploids than in haploids and when they are partially recessive in heterozygous diploids. The model predicts that effective dominance-a measure that accounts for these two effects-should be close to 0.5 in these species. It also predicts that diploids should have higher fitness than haploids on average. However, an appropriate statistical framework to conjointly investigate these predictions is currently lacking. In this study, we derive a new quantitative genetic model to test these predictions using fitness data of two haploid parents and their diploid offspring, and genome-wide genetic distance between haploid parents. We apply this model to the root-rot basidiomycete fungus Heterobasidion parviporum-a species where the heterokaryotic (equivalent to the diploid) phase is longer than the homokaryotic (haploid) phase. We measured two fitness-related traits (mycelium growth rate and the ability to degrade wood) in both homokaryons and heterokaryons, and we used whole-genome sequencing to estimate nuclear genetic distance between parents. Possibly due to a lack of power, we did not find that deleterious mutations were recessive or more deleterious when expressed during the heterokaryotic phase. Using this model to compare effective dominance among haploid-diploid species where the relative importance of the two phases varies should help better understand the evolution of haploid-diploid life cycles.


External validation of a 3-step falls prediction model in mild Parkinson's disease.
The 3-step falls prediction model (3-step model) that include history of falls, history of freezing of gait and comfortable gait speed <1.1 m/s was suggested as a clinical fall prediction tool in Parkinson's disease (PD). We aimed to externally validate this model as well as to explore the value of additional predictors in 138 individuals with relatively mild PD. We found the discriminative ability of the 3-step model in identifying fallers to be comparable to previously studies [area under curve (AUC), 0.74; 95 % CI 0.65-0.84] and to be better than that of single predictors (AUC, 0.61-0.69). Extended analyses generated a new model for prediction of falls and near falls (AUC, 0.82; 95 % CI 0.75-0.89) including history of near falls, retropulsion according to the Nutt Retropulsion test (NRT) and tandem gait (TG). This study confirms the value of the 3-step model as a clinical falls prediction tool in relatively mild PD and illustrates that it outperforms the use of single predictors. However, to improve future outcomes, further studies are needed to firmly establish a scoring system and risk categories based on this model. The influence of methodological aspects of data collection also needs to be scrutinized. A new model for prediction of falls and near falls, including history of near falls, TG and retropulsion (NRT) may be considered as an alternative to the 3-step model, but needs to be tested in additional samples before being recommended. Taken together, our observations provide important additions to the evidence base for clinical fall prediction in PD.


Key adhesion molecules are present on long podia extended by hematopoietic cells.
We recently reported that CD34(+) hematopoietic cells and the KG1a cell line extend long, thin podia. These podia can dynamically extend and retract, often adhere to the substrate, and appear to connect cells up to 300 microm apart. The surface receptors found on these podia have not been described. By using time-lapse fluorescent microscoscopy and immunostaining techniques, we describe a method for detecting surface receptors on these podia. This includes an in situ antibody staining procedure without fixing cells. We demonstrate, using CD34 selected mobilized peripheral blood cells and KG1a cells, that adhesion molecules known to play important roles in blood-cell migration and adhesion are present on these podia. These include: CD11a, CD18, CD29, CD34, CD45, CD49d, CD49e, and CD62L. Additionally, CD54 and CD44 were present on the podia extended by KG1a cells, but were not detectable on the primary CD34(+) cells. The integrin CD49d localized at the base of these podia in a time-dependent manner in KG1a cells. The frequency and morphology of these long podia on three myeloid leukemia-cell lines (KG1a, MV4-11, and AML-193) and a CD34-negative T-cell line (CEM) are also compared. KG1a and CEM cell lines extend long, dynamic podia that are similar to the podia on primary CD34(+) cells in morphology and adhesion molecule expression. The AML-193 and MV4-11 cell lines, however, did not extend these long podia. We describe a technique that provides a method of detecting surface receptors on thin cell membrane projections. These results support the likely role of these podia in cell migration and cell-cell communication.


Structural features of mammalian gonadotropins.
There are two species for which both pituitary and placental gonadotropins are readily available, humans and horses. The human gonadotropins are better characterized than equine gonadotropins. Nevertheless, the latter are very interesting because they provide exceptions to some of the general structure-function principles derived from studies on human and other mammalian gonadotropins. For example, separate genes encode the hLH beta and hCG beta subunits while a single gene encodes eLH beta and eCG beta. Thus, eCG and eLH differ only in their oligosaccharide moieties and eLH is the only LH that possesses the O-glycosylated C-terminal extension previously believed to be restricted to chorionic gonadotropins. Truncation experiments involving eLH beta and hCG beta have suggested the C-terminal extension has no effect on receptor binding. However, the largest of three eCG forms which differ only in the extent of O-glycosylation possessed reduced affinity for LH and FSH receptors. This result suggested that effects of O-glycosylation need to be considered when examining the glycosylation differences between eLH and eCG responsible for the 10-fold lower eCG receptor binding affinity compared with that of eLH. Contribution of alpha Asn56 N-linked oligosaccharides to the different biological activities of eLH and eCG has been evaluated following selective removal using peptide-N-glycanase digestion of native equine alpha-subunit preparations. Hormones-specific patterns of glycosylation were observed on alpha Asn56 of eLH, eFSH, and eCG. Removal of alpha Asn56 oligosaccharides increased the rate of subunit association, the extent of association, and receptor binding activity. Some unassociated alpha-subunit oligosaccharides were identified which may interfere with subunit association because they were more abundant in unassociated subunit oligosaccharide maps than in a total oligosaccharide map. This was most striking in the case of eCG alpha in which two minor peaks became the major oligosaccharide peaks detectable in the unassociated eCG alpha fraction following association with eLH beta and eFSH beta. The biological activities exhibited by hybrid hormones, eLH alpha reassociated with oLH beta and pLH beta, found to be greater than those of oLH and pLH provided an interesting exception to the general rule that the beta-subunit determines the potency of the heterodimer. LH receptor binding activities of eLH beta-chimeric ovine/equine alpha-subunits suggested that the equine alpha-subunit N-terminal domain may be responsible for this effect. Equine FSH has higher FSH receptor binding activity than human, ovine, and porcine FSH preparations. This probably results from two factors. First, the presence of the equine alpha-subunit promotes receptor binding as noted above. Second, the overall -2 charge of the eFSH beta determinant loop, which is less negative that the -3 observed in other species, results from the presence of an Asn residue at position 88 instead of Asp. This apparently facilitates binding to the FSH receptor.


A follow-up study of maternal expressed emotion toward children with Attention-Deficit/Hyperactivity Disorder (ADHD): relation with severity and persistence of ADHD and comorbidity.
Attention-deficit/hyperactivity disorder (ADHD) is associated with conflicted parent-child relationships. The underlying mechanisms of this association are not yet fully understood. We investigated the cross-sectional and longitudinal relationships between externalizing psychopathology in children with ADHD, and expressed emotion (EE; warmth and criticism) and psychopathology in mothers. In this 6-year follow-up study, 385 children with an ADHD combined subtype were included at baseline (mean, 11.5 years, 83.4% male), of which 285 children (74%) were available at follow-up (mean, 17.5 years, 83.5% male). At both time points, measures of child psychopathology (i.e., ADHD severity, oppositional, and conduct problems), maternal EE, and maternal psychopathology (i.e., ADHD and affective problems) were obtained. EE was not significantly correlated over time. At baseline, we found a nominally negative association (p ≤ .05) between maternal warmth and child ADHD severity. At follow-up, maternal criticism was significantly associated with child oppositional problems, and nominally with child conduct problems. Maternal warmth was nominally associated with child oppositional and conduct problems. These associations were independent of maternal psychopathology. No longitudinal associations were found between EE at baseline and child psychopathology at follow-up, or child psychopathology at baseline and EE at follow-up. The results support previous findings of cross-sectional associations between parental EE and child psychopathology. This, together with the finding that EE was not stable over 6 years, suggests that EE is a momentary state measure varying with contextual and developmental factors. EE does not appear to be a risk factor for later externalizing behavior in children with ADHD.