Advance telephone calls ahead of reminder questionnaires increase response rate in non-responders compared to questionnaire reminders only: The RECORD phone trial.
Postal questionnaires are simple and economical for collecting outcome data for randomised controlled trials (RCTs) but are prone to non-response. In the RECORD trial (a large pragmatic publicly funded RCT in UK) non-responders were sent a reminder and another questionnaire at 1 year, of which 40% were returned. In subsequent years we investigated the effect of an advance telephone call to non-responders on responses rate to reminder questionnaires and the next questionnaire 4 months later. Non-responders to annual questionnaires were randomised to receive a telephone call from the trial office ahead of the reminder questionnaire in addition to the usual reminder schedule (n=390) or to a control group that received the usual reminder schedule only (n=363). The primary outcome was response to the reminder questionnaire within 21 days; secondary outcomes were response to a questionnaire 4 months later; completeness of quality of life instruments; and the number of participants declining further follow-up. Results are presented as odds ratios from a logistic regression intention-to-treat (ITT) analysis and then percentage difference and 95% confidence intervals (CI) for both ITT and average treatment effect on the treated (ATT) analyses. The proportions that responded were 67.8% (265/390) in the intervention group compared to 62.5% (227/363) in the control group. The ITT estimate was a 5.4% increase (95% CI -1.4 to 12.2). Four months later percentages responding were 51.8% (202) and 42.7% (155). The ITT estimate was a 9.1% increase (95% CI 2.0 to 16.2). In the intervention group 12.3% (48/390) of participants were not telephoned because questionnaires were returned before the scheduled telephone call. ATT estimates adjusting for this were 6.2% (95% CI -1.6 to 14.0) and 10.4% (95% CI 2.2 to 18.5), respectively. The telephone call resulted in a slight increase in response to the reminder questionnaire, however at 4 months later the proportion in the telephoned group responding was greater. This study suggests that pre-notification telephone calls may only be worthwhile if further questionnaires are to be sent out soon after reminder questionnaires. Current Clinical Trials ISRCTN51647438.


Phosphotungstic acid functionalized silica nanocomposites with tunable bicontinuous mesoporous structure and superior proton conductivity and stability for fuel cells.
A novel proton exchange membrane using phosphotungstic acid (HPW) as proton carrier and cubic bicontinuous Ia3d mesoporous silica (meso-silica) as framework material is successfully developed as proton exchange membranes for fuel cells. Meso-silica is functionalized by 80wt% HPW using a vacuum impregnation method. The HPW-functionalized meso-silica (HPW-meso-silica) nanocomposites are characterized by transmission electron microscopy (TEM), small angle X-ray scattering (SAXS), N(2) adsorption/desorption isotherms, thermogravimetric analysis (TGA), water uptake and four-probe conductivity. The results show that the mesoporous structure of silica hosts can be altered by the hydrothermal temperature. Conductivity measurements indicate that meso-silica host with pore diameter of 5.0 nm has the highest proton conductivity of 0.11 S cm(-1) at 80 °C and 100% relative humidity (RH) with an activation energy of ∼14 kJ mol(-1) and better stability as compared to that with large mesopores. The proton conductivity and performance of HPW-meso-silica nanocomposites also increase with the RH, but it is far less sensitive to RH changes as compared to conventional perfluorosulfonic acid (PFSA) polymers such as Nafion. The maximum power density of the cell with HPW-meso-silcia nanocomposite membranes is 221 mW cm(-2) at 80 °C and 100% RH and decreases to 171 mW cm(-2) when RH is reduced to 20%, a 20% decrease in power output. In the case of a cell with Nafion 115 membranes, the decrease in power density is 95% under identical test conditions. The results demonstrate that the HPW-meso-silica nanocomposite has an exceptionally high water retention capability and is a promising proton exchange membrane material for fuel cells operating at reduced humidity and elevated temperatures.


Predicting the required number of training samples.
In this paper a criterion which measures the quality of the estimate of the covariance matrix of a multivariate normal distribution is developed. Based on this criterion, the necessary number of training samples is predicted. Experimental results which are used as a guide for determining the number of training samples are included.


Long-term remission with rituximab in a patient with severe hepatitis C virus-induced mixed cryoglobulinemia.
Mixed cryoglobulinemia (MC) is a small-vessel systemic vasculitis characterized by the presence of cryoglobulins, immunoglobulin complexes that precipitate at low temperatures ( < 37 ºC) inducing the inflammatory process. The most frequent etiology is hepatitis C infection (HCV) (1). Rituximab (RTX), an anti-CD20 monoclonal antibody, has recently emerged as the treatment of choice for severe MC (2). We present a case of severe hepatitis C virus-induced MC that was controlled and maintained in remission with RTX for 26 months, a remarkable prolonged period of time.


Physical polymer surface modification methods and applications in food packaging polymers.
Continued innovations in the polymer industry have made polymer surface modification methods a subject of intense research. The importance and necessity of surface modification of plastics are explained, and the advantages of physical surface treatments over the less-sophisticated chemical methods are outlined. Currently available physical surface modification methods for food packaging polymers are reviewed from the food packaging perspective. These physical surface modification methods include flame, corona discharge, UV, gamma-ray, electron beam, ion beam, plasma, and laser treatments. The principle of operation of each method is briefly described, and the advantages and disadvantages of each technique are cited. The extent to which each of these methods can produce the specific modifications desired is discussed. Furthermore, the effects of each treatment on barrier, mechanical, and adhesion properties of food packaging polymers are also examined. Finally, an overview of economic aspects of sophisticated surface modification techniques, including ion beam, plasma, and laser treatments, is presented.


Estimation of pedestrian level UV exposure under trees.
Trees influence the amount of solar UV radiation that reaches pedestrians. A three-dimensional model was developed to predict the ultraviolet-B (UV-B) irradiance fields in open-tree canopies where the spacing between trees is equal to or greater than the width of individual tree crowns. The model predicted the relative irradiance (fraction of above-canopy irradiance) under both sunlit and shaded conditions under clear skies with a mean bias error of less than 0.01 and a root mean square error of 0.07. Both model and measurements showed that the locations people typically perceive as shady, low-irradiance locations in the environment can actually have significant UV-B exposure (40-60% of that under direct sunlight). The relationship of tree cover in residential neighborhoods to erythemal UV-B exposure for children and adults was modeled for the 4 h around noon in June and July. Results showed that human exposures (on the horizontal) in cities located at 15 and 30 degrees latitudes are nearly identical. For latitudes between 15 and 60 degrees, ultraviolet protection factors (UPF) were less than 2 for less than 50% tree cover. A UPF of 10 was possible at all latitudes for tree cover of 90%.


[3 linkage groups of the genes of riboflavin biosynthesis in Escherichia coli].
Using transposon Tn5 inactivation technology a collection of Escherichia coli mutants defective in riboflavine biosynthesis was obtained. All mutations were distributed within three linkage groups. With the help of P1-transduction mapping, group I mutations (ribA locus) were localized near cysB locus (28 min of the standard 100 min E. coli map) and mutations of group II (ribB locus) were mapped near tolC locus (66 min). The location of group III mutations was approximately determined by the F' complementation analysis: this linkage group lies in the region of 56-60 min of the E. coli map.


Expanding dental education partnerships beyond the four walls.
The increasing complexities of health care that dental graduates must be equipped to handle require schools to develop new models of education in order to address these intricacies. To meet these challenges, it is the school's responsibility to provide an environment that fosters discovery and scholarly activity, embraces evidence-based philosophies, encourages partnerships with other units on campus and the community, including the global community, and recognizes the richness of diversity in both our human resources and our thinking. Beyond new curriculum initiatives within our school, we recognized the need to build strong partnerships outside our four walls in order to respond to the challenges confronting us. Four such notable recent initiatives at the University of Washington School of Dentistry discussed in this article are Regional Initiatives in Dental Education, the Center for Pediatric Dentistry: Program in Early Childhood Oral Health, Northwest PRECEDENT (Practice-based REsearch Collaborative in Evidence-based DENTistry), and Alaska Native Dental Health Aide Therapist Program. These partnerships focus on new models to improve access to care and to enhance the impact of research on evidence-based practice. These are examples of the many opportunities for us to act collectively in creating new models that ensure our graduates have the knowledge, attitudes, and skills to be competent oral health care professionals.


Development of a novel squalene/α-tocopherol-based self-emulsified nanoemulsion incorporating Leishmania peptides for induction of antigen-specific immune responses.
Vaccination has emerged as the most effective strategy to confront infectious diseases, among which is leishmaniasis, that threat public health. Despite laborious efforts there is still no vaccine for humans to confront leishmaniasis. Multi-epitope protein/peptide vaccines present a number of advantages, however their use along with appropriate adjuvants that may also act as antigen carriers is considered essential to overcome subunit vaccines' low immunogenicity. In the present study, a stable self-emulsified nanoemulsion was developed and double-adjuvanted with squalene and α-tocopherol. The prepared nanoemulsion droplets exhibited low cytotoxicity in a certain range of concentrations, while they were efficiently taken up by macrophages and dendritic cells in vitro as well as in vivo in secondary lymphoid organs. To further characterize nanoformulation's potent antigen delivery capability, three multi-epitope Leishmania peptides were incorporated into the nanoemulsion. Peptide encapsulation resulted in dendritic cells' functional differentiation characterized by elevated levels of maturation markers and intracellular cytokine production. Intramuscular administration of the nanoemulsion incorporating Leishmania peptides induced antigen-specific spleen cell proliferation as well as elicitation of CD4+ central memory cells, supporting the potential of the developed nanoformulation to successfully act also as an antigen delivery vehicle and thus encouraging further preclinical studies on its vaccine candidate potency.


[Melatonin in the combined treatment of cardiovascular diseases].
The aim of the work was to assess effectiveness of melatonin monotherapy (MT) and combined treatment (CT) in aged patients with arterial hypertension (AH) and coronary heart disease (CHD). A group of 170 patients (mean age 64 years) included 61 ones with CHD and 100 with AH. ECG, AP, and echoCG monitoring was performed before and 21 days after treatment along with clinical examination and evaluation of oxidative and antioxidative activities in erythrocytes. Treatment efficiency was compared in the study and randomized control groups. The data obtained suggest antihypertensive action of MT. CT with melatonin and antihypertensive drugs had better outcome than standard therapy in control groups. Inclusion of melatonin in CT of CHD produced marked anti-ischemic and anti-anginal effects and normalized oxidant/antioxidant balance. It is concluded that melatonin should be an important component of MT and CT of cardiovascular diseases.


A Platform Approach to Protein Encapsulates with Controllable Surface Chemistry.
The encapsulation of proteins into core-shell structures is a widely utilised strategy for controlling protein stability, delivery and release. Despite the recognised utility of these microstructures, however, core-shell fabrication routes are often too costly or poorly scalable to allow for industrial translation. Furthermore, many scalable routes rely upon emulsion-techniques implicating denaturing or environmentally harmful organic solvents. Herein, we investigate core-shell protein encapsulation through single-feed, aqueous spray drying: a cheap, industrially ubiquitous particle-formation technology in the absence of organic solvents. We show that an excipient's preference for the surface of the spray dried particle is well-predicted by its hydrodynamic diameter (Dh) under relevant feed buffer conditions (pH and ionic strength) and that the predictive power of Dh is improved when measured at the spray dryer outlet temperature compared to room temperature (R2 = 0.64 vs. 0.59). Lastly, we leverage these findings to propose an adaptable design framework for fabricating core-shell protein encapsulates by single-feed aqueous spray drying.


Negative Temperature Coefficient of Resistance in Bismuth I.
Measurements of the electrical resistance have been made on bismuth 1 between 15 and 35 kilobars at temperatures between 77.4 degrees and 120 degrees K. Above about 150 degrees K, the temperature coefficient of resistance is positive, as in a metal; below 150 degrees K, the coefficient becomes negative, as is characteristic of semiconductors. On the basis that bismuth is a semiconductor, the energy gap, calculated by the exponential resistance formula, is 0.006 ev at 15 kb with a steady rise to 0.018 ev at 35 kb. At higher pressures, bismuth I is transformed into a metallic modification with the normal temperature dependence of the resistance. The energy gap in bismuth I is not visible at room temperature because thermal excitation populates the conduction band and metallic behavior is the result. From available evidence the observed behavior is due to an energy gap rather than to a decrease in carrier mobility.


An improved algorithm for determining HIV type 1 subtypes in a primary laboratory in Uganda.
A pilot study was undertaken with the objective of developing a simple, economical, and efficient algorithm through which to subtype HIV-1 in a large epidemiological cohort study in Uganda. A peptide enzyme immunoassay (PEIA) employing both V3 and gp41 regions and a heteroduplex mobility assay (HMA) were evaluated in comparison with DNA sequencing. Of 146 samples selected, 115 (79%) were successfully sequenced. Taking sequence data as the "gold standard," other assays were compared with these data. The HMA correctly identified 95 (83%) of the samples, and only 1 sample was wrongly identified. The V3 PEIA alone and in combination with gp41 peptides correctly identified 76 and 78% of the samples, respectively; however, the number of wrongly identified samples was four times less with the combination compared with V3 peptides alone (4 versus 16%). The sensitivity, specificity, and positive and negative predictive values for serotype A and D samples were greater for the combination than V3 peptides alone. We have described a new algorithm to segregate subtypes A and D. This algorithm uses the two peptide assays followed by HMA and then DNA sequencing for untypable samples, giving an accuracy of 95% at a cost of 37 and 21% for consumables compared with subtyping all the samples by HMA or DNA sequencing, respectively. This proposed approach is suitable for epidemiological studies in Uganda and other regions with a predominance of A and D subtypes.


Development, Diversity, and Function of Dendritic Cells in Mouse and Human.
The study of murine dendritic cell (DC) development has been integral to the identification of specialized DC subsets that have unique requirements for their form and function. Advances in the field have also provided a framework for the identification of human DC counterparts, which appear to have conserved mechanisms of development and function. Multiple transcription factors are expressed in unique combinations that direct the development of classical DCs (cDCs), which include two major subsets known as cDC1s and cDC2s, and plasmacytoid DCs (pDCs). pDCs are potent producers of type I interferons and thus these cells are implicated in immune responses that depend on this cytokine. Mouse models deficient in the cDC1 lineage have revealed their importance in directing immune responses to intracellular bacteria, viruses, and cancer through the cross-presentation of cell-associated antigen. Models of transcription factor deficiency have been used to identify subsets of cDC2 that are required for T helper (Th)2 and Th17 responses to certain pathogens; however, no single factor is known to be absolutely required for the development of the complete cDC2 lineage. In this review, we will discuss the current state of knowledge of mouse and human DC development and function and highlight areas in the field that remain unresolved.


Sertoli cell tumour in a pet rabbit (Oryctolagus cuniculus): histological and immunohistochemical characterization.
The present study describes a case of a spontaneous, unilateral Sertoli cell tumour (SCT) in a 6-year-old pet rabbit. The rabbit was presented with a palpable, unilateral, subcutaneous left inguinal mass, consistent with the suspected clinical diagnosis of neoplasia developing within the retained testis in the inguinal canal. The intrascrotal contralateral testis was palpable, but reduced in volume. The rabbit underwent orchiectomy and both the testes were collected, formalin-fixed, and submitted for histopathological examination. Microscopically, the enlarged testis was effaced by an intratubular SCT in which numerous intratubular microliths were evident. The contralateral testis was severely atrophied. Immunohistochemical stains showed neoplastic Sertoli cells that were diffusely positive for vimentin and anti-Müllerian-Hormone and multifocally positive for cytokeratins and desmin. Eighteen months after the surgery, the rabbit showed no clinical signs of disease. This is the first report of a spontaneously occurring rabbit SCT histologically described and immunohistochemically investigated.


Determination of initiation of DNA replication before and after nuclear formation in Xenopus egg cell free extracts.
Xenopus egg extracts prepared before and after egg activation retain M- and S-phase specific activity, respectively. Staurosporine, a potent inhibitor of protein kinase, converted M-phase extracts into interphase-like extracts that were capable of forming nuclei upon the addition of sperm DNA. The nuclei formed in the staurosporine treated M-phase extract were incapable of replicating DNA, and they were unable to initiate replication upon the addition of S-phase extracts. Furthermore, replication was inhibited when the staurosporine-treated M-phase extract was added in excess to the staurosporine-treated S-phase extract before the addition of DNA. The membrane-depleted S-phase extract supported neither nuclear formation nor replication; however, preincubation of sperm DNA with these extracts allowed them to form replication-competent nuclei upon the addition of excess staurosporine-treated M-phase extract. These results demonstrate that positive factors in the S-phase extracts determined the initiation of DNA replication before nuclear formation, although these factors were unable to initiate replication after nuclear formation.


The new WHO classification for essential thrombocythemia calls for revision of available evidences.
In the 2016 revised classification of myeloproliferative neoplasms pre-fibrotic primary myelofibrosis (pre-PMF) was recognized as a separate entity, distinct from essential thrombocythemia (ET). Owing that the majority of cases falling in the pre-PMF category were previously diagnosed as ET, one may question about the need to re-evaluate the results of epidemiologic, clinical, and molecular studies, and the results of clinical trials in the two entities. Based on a critical review of recently published studies, pre-PMF usually presents with a distinct clinical and hematological presentation and higher frequency of constitutional symptoms. JAK2V617F and CALR mutations in pre-PMF patients are superimposable to ET, whereas non-driver high-risk mutations are enriched in pre-PMF compared with ET. Thrombosis is not significantly different, whereas bleeding is more frequent in pre-PMF. Median survival is significantly shorter in pre-PMF and 10-year cumulative rates progression to overt myelofibrosis is 0-1% vs. 10-12%, and leukemic transformation is 1-2% vs. 2-6%, in ET and pre-fibrotic-PMF, respectively. Most patients fall in the lower prognostic IPSS group in which observation alone can be recommended. Patients at intermediate risk may require a symptom-driven treatment for anemia, splenomegaly or constitutional symptoms while cytoreductive drugs are indicated in the high-risk category.


Oral steroids in initial treatment of acute sciatica.
Many physicians use prednisone to treat acute sciatica with the hope of speeding recovery. There is little clinical evidence to support this practice. Our objective was to determine whether early administration of oral prednisone affects parameters related to recovery from acute sciatica. In this double-blind, controlled clinical trial, 27 patients were sequentially assigned to receive either a 9-day tapering course of prednisone (n = 13) or placebo (n = 14) within 1 week of developing sciatic symptoms. Patients and investigators were blinded to the drug administered. Follow-up assessment was done weekly for 1 month and then monthly for 5 months. Prednisone and control groups showed no statistically significant differences in physical findings, use of nonsteroidal anti-inflammatory drugs or narcotic medications, or rates of patients returning to work at any time interval studied. Compared with controls, patients who received prednisone had more rapid rates of improvement from baseline in pain, mental well-being, and disability scores. These changes were subtle but statistically significant. Patients who received prednisone tended to receive fewer epidural injections for pain. Early administration of oral steroid medication in patients with acute sciatica had no significant effect on most parameters studied. It did, however, lead to slightly more rapid rates of improvement in pain, mental well-being, and disability scores. The impact of oral steroids on other outcomes is suggested by this study, but its small sample size limited its statistical power.


OneFlorida Clinical Research Consortium: Linking a Clinical and Translational Science Institute With a Community-Based Distributive Medical Education Model.
Developing a national pragmatic clinical trial infrastructure is central to understanding the effectiveness of interventions applied under usual conditions and where people receive health care. To address this challenge, three Florida universities-the University of Florida Clinical and Translational Science Institute, Florida State University (with its community-based distributive medical education model), and the University of Miami-created (2010-2013) a statewide consortium, the OneFlorida Clinical Research Consortium, to support the conduct of pragmatic clinical trials and provide mentored research experiences for medical and graduate students in real-world practice settings. OneFlorida has four programs, which report to a steering committee with membership from each partner, community members, and the state Medicaid agency and Department of Health to ensure shared governance. The Clinical Research Program provides support to conduct research in the network and uses champions to engage community clinicians. The Citizen Scientist Program has community members who provide input on health topics of importance to them, study design, recruitment and retention strategies, and the interpretation of findings. The Data Trust Program contains electronic health record and health care claims data for 10.6 million Floridians. The Minority Education Program, in collaboration with three historically black colleges and universities, offers minority junior faculty mentoring in pragmatic clinical trials and implementation science. OneFlorida has implemented 27 studies with diverse patient populations and in diverse community practice settings. To identify evidence-based best practices from the clinical trials conducted in the network, foster their implementation, and expand research training opportunities.


Economic burden of blindness in India.
Economic analysis is one way to determine the allocation of scarce resources for health-care programs. The initial step in this process is to estimate in economic terms the burden of diseases and the benefit from interventions for prevention and treatment of these diseases. In this paper, the direct and indirect economic loss due to blindness in India is calculated on the basis of certain assumptions. The cost of treating cataract blindness in India is estimated at current prices. The economic burden of blindness in India for the year 1997 based on our assumptions is Rs. 159 billion (US$ 4.4 billion), and the cumulative loss over lifetime of the blind is Rs. 2,787 billion (US$ 77.4 billion). Childhood blindness accounts for 28.7% of this lifetime loss. The cost of treating all cases of cataract blindness in India is Rs. 5.3 billion (US$ 0.15 billion). Similar estimates for causes of blindness other than cataract have to be made in order to develop a comprehensive approach to deal with blindness in India.


Effects of 'Five freedoms' environmental enrichment on the welfare of calves reared indoors.
We provided comfortable resting areas in pens and tools for self-grooming and suppressing aggressive behavior for environmental enrichment in Japanese black and Japanese Shorthorn calves. At the start of the experiment, the black calves (n = 10) were 164 days old and 138 kg in weight, and the Japanese Shorthorn calves (n = 10) were 176 days old and 164 kg in weight. Calves of each breed were divided into two groups (the enrichment group and the non-treatment group), and each group was housed in a 4 m x 7.2 m pen. The effect of enrichment on calf welfare was estimated on the basis of daily weight gain and behavior. The brush stimulated the grooming behavior in calves although the mean time of using the brush decreased during the experiment. The partition of the feed trough and the wooden wall in the pen decreased the agonistic behavior in Japanese Shorthorn calves (P < 0.01) and increased the affiliative behavior in Japanese black calves (P = 0.08). The cleanliness of the bedding increased sternum lying and lying with the head touching the flank or ground in Japanese black calves and decreased the standing resting behavior in both breeds (P < 0.05 in all the cases). However, the treatments did not have an effect on daily weight gain. We concluded that the treatments would improve the welfare of calves and function as environmental enrichments.


Soliciting strategies for developing cell-based reference materials to advance mesenchymal stromal cell research and clinical translation.
The mesenchymal stromal cell (MSC) field continues to rapidly progress with a number of clinical trials initiated and completed, with some reported successes in multiple clinical indications, and a growing number of companies established. The field, nevertheless, faces several challenges. Persistent issues include the definition of a MSC and comparability between MSC preparations. This is because of inherent cell heterogeneity, the absence of markers that are unique to MSCs, and the difficulty in precisely defining them by developmental origin. Differences in the properties of MSCs also depend on the site of tissue harvest, phenotypic and genotypic characteristics of the donor and the isolation, and storage and expansion methods used. These differences may be sufficient to ensure that attributes of the final MSC product could differ in potentially significant ways. Since there are currently no gold standards, we propose using a reference material to establish methods of comparability among MSC preparations. We suggest four possible "ruler scenarios" and a method for global distribution. We further suggest that critical to establishing a reference material is the need to define protocols for comparing cells. The main purpose of this article is to solicit input in establishing a consensus-based comparison. A comparative approach will be critical to all stages of translation to better clarify mechanisms of MSC actions, define an optimal cell manufacturing process, ensure best practice clinical investigations, extend the use of an MSC product for new indications, protect an MSC product from imitators, and develop uniform reimbursement policies. Importantly, a reference material may enable a consensus on a practical definition of MSCs.


Severe arterial injury heals with a complex clonal structure involving a large fraction of surviving smooth muscle cells.
Smooth muscle cell (SMC) lineage cells in atherosclerosis and flow cessation-induced neointima are oligoclonal, being recruited from a tiny fraction of medial SMCs that modulate and proliferate. The present study aimed to investigate the clonal structure of SMC lineage cells healing more severe arterial injury. Arterial injury (wire, stretch, and partial ligation) was inflicted on the right carotid artery in mice with homozygous, SMC-restricted, stochastically recombining reporter transgenes that produced mosaic expression of 10 distinguishable fluorescent phenotypes for clonal tracking. Healed arteries and contra-lateral controls were analyzed after 3 weeks. Additional analysis of cell death and proliferation after injury was performed in wildtype mice. The total number of SMC lineage cells in healed arteries was comparable to normal arteries but comprised significantly fewer fluorescent phenotypes. The population had a complex, intermixed, clonal structure. By statistical analysis of expected versus observed fractions of fluorescent phenotypes and visual inspection of coherent groups of same-colored cells, we concluded that >98% of SMC lineage cells in healed arteries belonged to a detectable clone, indicating that nearly all surviving SMCs after severe injury at some point undergo proliferation. This was consistent with serial observations in the first week after injury, which showed severe loss of medial cells followed by widespread proliferation. After severe arterial injury, many surviving SMCs proliferate to repair the media and form a neointima. This indicates that the fraction of medial SMCs that are mobilized to repair arteries increases with the level of injury.


Starting patients with type 2 diabetes on insulin therapy using once-daily injections of biphasic insulin aspart 70/30, biphasic human insulin 70/30, or NPH insulin in combination with metformin.
Transitioning safely to insulin therapy when oral antidiabetic agents fail to provide adequate glycemic control is a critical aspect of care for the patient with type 2 diabetes mellitus (T2DM). We evaluated the clinical effectiveness of starting patients on a relatively simple regimen of once-daily injections of either biphasic insulin aspart 70/30 (10 min before dinner), NPH insulin (at 10 p.m.), or biphasic human insulin 70/30 (30 min before dinner) in combination with metformin. Enrolled patients had T2DM and inadequate glycemic control (AlC>/=7.5%) on a previous regimen of metformin as monotherapy or in combination with a sulphonylurea. One hundred and forty (140) patients received metformin monotherapy for 4 weeks followed by 12 weeks of combination treatment with metformin and once-daily insulin injections. AlC levels decreased from baseline by 1.1-1.3% for patients in each of the three treatment groups. Overall, FPG values decreased from baseline by 31% (biphasic insulin aspart), 37% (NPH insulin), and 28% (biphasic human insulin). Subjects whose final FPG level was <126 mg/dl experienced the largest decreases in AlC values (-2.3%, -1.9%, -1.8%, respectively). All three treatment regimens were well tolerated. The results indicate that patients with T2DM can safely and effectively begin insulin therapy using once-daily injections of biphasic insulin aspart 70/30, biphasic human insulin 70/30, or NPH insulin in combination with metformin.


Hb Aalesund (HBA2: c.400_406delAGCACCG), an Unstable α-Globin Variant Found in a Norwegian Patient Causing Moderate Hemolytic Anemia and Falsely High Hb A1c Using Ion Exchange High Performance Liquid Chromatography.
A new unstable hemoglobin (Hb) variant, named Hb Aalesund, was detected during Hb A1c measurement in a patient with a nearly compensated hemolytic anemia. Sequencing of the α-globin genes revealed a 7 bp deletion in exon 3 of the HBA2 gene (HBA2: c.400_406delAGCACCG) (NM_000517.4) causing a frameshift and a premature termination codon (PTC) two positions downstream. Apparently, the transcript bypassed nonsense-mediated decay (NMD), and a truncated protein was translated. The unstable Hb variant presumably underwent rapid denaturation, as heterozygosity of Hb Aalesund was associated with mild hemolytic anemia. In addition, the Hb variant interfered with Hb A1c measurement by cation exchange high performance liquid chromatography (HPLC), causing a falsely high Hb A1c result when using the Bio-Rad D10™ Hemoglobin Analyzer fast Hb A1c Program.


Microsatellite marker development in the crop wild relative Linum bienne using genome skimming.
Nuclear microsatellite markers were developed for Linum bienne, the sister species of the crop L. usitatissimum, to provide molecular genetic tools for the investigation of L. bienne genetic diversity and structure. Fifty microsatellite loci were identified in L. bienne by means of genome skimming, and 44 loci successfully amplified. Of these, 16 loci evenly spread across the L. usitatissimum reference nuclear genome were used for genotyping six L. bienne populations. Excluding one monomorphic locus, the number of alleles per locus ranged from two to 12. Four out of six populations harbored private alleles. The levels of expected and observed heterozygosity were 0.076 to 0.667 and 0.000 to 1.000, respectively. All 16 loci successfully cross-amplified in L. usitatissimum. The 16 microsatellite loci developed here can be used for population genetic studies in L. bienne, and 28 additional loci that successfully amplified are available for further testing.


Cornea organoids from human induced pluripotent stem cells.
The cornea is the transparent outermost surface of the eye, consisting of a stratified epithelium, a collagenous stroma and an innermost single-cell layered endothelium and providing 2/3 of the refractive power of the eye. Multiple diseases of the cornea arise from genetic defects where the ultimate phenotype can be influenced by cross talk between the cell types and the extracellular matrix. Cell culture modeling of diseases can benefit from cornea organoids that include multiple corneal cell types and extracellular matrices. Here we present human iPS cell-derived organoids through sequential rounds of differentiation programs. These organoids share features of the developing cornea, harboring three distinct cell types with expression of key epithelial, stromal and endothelial cell markers. Cornea organoid cultures provide a powerful 3D model system for investigating corneal developmental processes and their disruptions in diseased conditions.


Bromoperoxidase activity of vanadate-substituted acid phosphatases from Shigella flexneri and Salmonella enterica ser. typhimurium.
Vanadium haloperoxidases and the bacterial class A nonspecific acid phosphatases have a conserved active site. It is shown that vanadate-substituted recombinant acid phosphatase from Shigella flexneri (PhoN-Sf) and Salmonella enterica ser. typhimurium (PhoN-Se) in the presence of H2O2 are able to oxidize bromide to hypobromous acid. Vanadate is essential for this activity. The kinetic parameters for the artificial bromoperoxidases have been determined. The Km value for H2O2 is about the same as that for the vanadium bromoperoxidases from the seaweed Ascophyllum nodosum. However, the Km value for Br- is about 10-20 times higher, and the turnover values of about 3.4 min-1 and 33 min-1 for PhoN-Sf and PhoN-Se, respectively, are much slower, than those of the native bromoperoxidase. Thus, despite the striking similarity in the active-site structures of the vanadium haloperoxidases and the acid phosphatase, the turnover frequency is low, and clearly the active site of acid phosphatases is not optimized for haloperoxidase activity. Like the native vanadium bromoperoxidase, the vanadate-substituted PhoN-Sf and PhoN-Se catalyse the enantioselective sulfoxidation of thioanisole.


Psychosexual therapy for delayed ejaculation based on the Sexual Tipping Point model.
The Sexual Tipping Point(®) (STP) model is an integrated approach to the etiology, diagnosis and treatment of men with delayed ejaculation (DE), including all subtypes manifesting ejaculatory delay or absence [registered trademark owned by the MAP Educational Fund, a 501(c)(3) public charity]. A single pathogenetic pathway does not exist for sexual disorders generally and that is also true for DE specifically. Men with DE have various bio-psychosocial-behavioral & cultural predisposing, precipitating, maintaining, and contextual factors which trigger, reinforce, or worsen the probability of DE occurring. Regardless of the degree of organic etiology present, DE is exacerbated by insufficient stimulation: an inadequate combination of "friction and fantasy". High frequency negative thoughts may neutralize erotic cognitions (fantasy) and subsequently delay, ameliorate, or inhibit ejaculation, while partner stimulation (friction) may prove unsatisfying. Assessment requires a thorough sexual history including inquiry into masturbatory methods. Many men with DE engage in an idiosyncratic masturbatory style, defined as a masturbation technique not easily duplicated by the partner's hand, mouth, or vagina. The clinician's most valuable diagnostic tool is a focused sex history (sex status). Differentiate DE from other sexual problems and review the conditions under which the man can ejaculate. Perceived partner attractiveness, the use of fantasy during sex, anxiety-surrounding coitus and masturbatory patterns require meticulous exploration. Identify important DE causes by juxtaposing an awareness of his cognitions and the sexual stimulation experienced during masturbation, versus a partnered experience. Assist the man in identifying behaviors that enhance immersion in excitation and minimize inhibiting thoughts, in order to reach ejaculation in his preferred manner. Discontinuing, reducing or altering masturbation is often required, which evokes patient resistance. Coaching tips are offered on how to ensure adherence to this suspension, manage resistance and facilitate success. Depending on motivation level, masturbation interruption may be compromised and negotiated. Encourage a man who continues to masturbate to alter style ("switch hands") and to approximate the stimulation likely to be experienced with his partner. Success will require most men to be taught to learn bodily movements and fantasies that approximate the thoughts and sensations experienced in masturbation. Fertility issues, as well as patient/partner anger are important causational factors, which often require individual and/or conjoint consultation. Drug treatment would benefit men particularly with severe DE, regardless of concomitant psychosocial-behavioral and cultural complications. When and if a safe effective medication for DE becomes available, this author's transdisciplinary perspective supports appropriate medication use when integrated with counseling. This approach emphasizes the utility of a biopsychosocial-cultural perspective combined with special attention to the patient's narrative. Treatment is patient-centered, holistic and integrates a variety of therapies as needed.


Effect of combined heat, ozonation and ultraviolet irradiation (VasoCare) on heat shock protein expression by peripheral blood leukocyte populations.
The re-administration of whole blood subjected to heat, ozonation and ultraviolet irradiation (VasoCare therapy) has been shown to elicit clinical benefits in individuals with vascular disease. Given that these stressors induce heat shock protein (Hsp) expression and that heat shock protein reactivity is implicated in the pathogenesis of vascular disease, this study assessed the effect of VasoCare on intracellular expression of Hsp60 and Hsp70 by treated peripheral blood leukocytes. Contrary to expectations, VasoCare induced a significant reduction (approximately 40%) in the proportion of peripheral blood mononuclear cells expressing intracellular Hsp60 and Hsp70, whereas it had no effect on heat shock protein expression by peripheral blood neutrophils. Cell surface heat shock protein expression was not detectable. The reduced expression of Hsp60 by mononuclear cells was concomitant with an increase in the levels of Hsp60 in treated plasma. Although the mechanism underlying the clinical effectiveness of VasoCare therapy has yet to be established, it may be that re-administration of treated blood or soluble factors derived therefrom modifies in vivo immune responsiveness to heat shock proteins or associated molecules.


Structural organization of signalling to and from IP3 receptors.
In the 30 years since IP3 (inositol 1,4,5-trisphosphate) was first shown to release Ca2+ from intracellular stores, the importance of spatially organized interactions within IP3-regulated signalling pathways has been universally recognized. Recent evidence that addresses three different levels of the structural determinants of IP3-evoked Ca2+ signalling is described in the present review. High-resolution structures of the N-terminal region of the IP3R (IP3 receptor) have established that the two essential phosphate groups of IP3 bind to opposite sides of the IP3-binding site, pulling its two domains together. This conformational change is proposed to disrupt an interaction between adjacent subunits within the tetrameric IP3R that normally holds the channel in a closed state. Similar structural changes are thought to allow gating of ryanodine receptors. cAMP increases the sensitivity of IP3Rs and thereby potentiates the Ca2+ signals evoked by receptors that stimulate IP3 formation. We speculate that both IP3 and cAMP are delivered to IP3Rs within signalling junctions, wherein the associated IP3Rs are exposed to a saturating concentration of either messenger. The concentration-dependent effects of extracellular stimuli come from recruitment of junctions rather than from a graded increase in the activity of individual junctions. IP3Rs within 'IP3 junctions' respond directly to receptors that stimulate phospholipase C, whereas extra-junctional IP3Rs are exposed to suboptimal concentrations of IP3 and open only when they are sensitized by cAMP. These results highlight the importance of selective delivery of diffusible messengers to IP3Rs. The spatial organization of IP3Rs also allows them to direct Ca2+ to specific intracellular targets that include other IP3Rs, mitochondria and Ca2+-regulated channels and enzymes. IP3Rs also interact functionally with lysosomes because Ca2+ released by IP3Rs, but not that entering cells via store-operated Ca2+ entry pathways, is selectively accumulated by lysosomes. This Ca2+ uptake shapes the Ca2+ signals evoked by IP3 and it may regulate lysosomal behaviour.


Whole exome sequencing identifies a novel mutation in ASPM and ultra-rare mutation in CDK5RAP2 causing Primary microcephaly in consanguineous Pakistani families.
Primary Microcephaly (MCPH) is a rare neurogenetic disease, manifesting congenitally reduced head circumference and non-progressive intellectual disability (ID). To date, twenty-eight genes with biallelic mutations have been reported for this disorder. The study aimed for molecular genetic characterization of Pakistani families segregating MCPH. We studied two unrelated consanguineous families (family A and B) presenting >2 patients with diagnostic symptoms of MCPH, born to asymptomatic parents. We employed whole-exome sequencing (WES) of probands to find putative causal mutations. The candidate variants were further confirmed and analyzed for co-segregation by Sanger sequencing of all available members of each family. This study was conducted at Government College University, Faisalabad, Pakistan, and Cologne Center for Genomics (CCG), University of Cologne, Germany; during 2017-2020. We identified a novel homozygous variant c.10097_10098delGA, p.(Gly3366Glufs*19) in exon 26 of ASPM gene in family A which presents with moderate intellectual disability, speech impairment, visual abnormalities, seizures, and ptyalism. Family B was found to segregate nonsense, homozygous variant c.448C>T p.(Arg150*) in CDK5RAP2. The patients also exhibited mild to severe seizures without ptyalism that has not been previously reported in patients with mutations in the CDK5RAP2 gene. We report a novel mutation in ASPM and ultra-rare mutation in the CDK5RAP2 gene, both causing primary microcephaly. The study expands the mutational spectrum of the ASPM gene to 212, and also adds to the clinical spectrum of CDK5RAP2 mutations. It also demonstrated the utility of WES in the investigation and genetic diagnosis of genetically heterogeneous disorders like MCPH. These findings would aid in diagnostic and preventive strategies including carrier screening, cascade testing, and genetic counselling.


Abnormal lacuno-canalicular network and negative correlation between serum osteocalcin and Cobb angle indicate abnormal osteocyte function in adolescent idiopathic scoliosis.
Adolescent idiopathic scoliosis (AIS) is a prevalent spinal deformity occurring during peripubertal growth period that affects 1-4% of adolescents globally without clear etiopathogenetic mechanism. Low bone mineral density is an independent and significant prognostic factor for curve progression. Currently, the cause underlying low bone mass in AIS remains elusive. Osteocytes play an important role in bone metabolism and mineral homeostasis, but its role in AIS has not been studied. In the present study, iliac bone tissues were harvested from 21 patients with AIS (mean age of 14.3 ± 2.20 yr old) with a mean Cobb angle of 55.6 ± 10.61° and 13 non-AIS controls (mean age of 16.5 ± 4.79 yr old) intraoperatively. Acid-etched scanning electron microscopy (SEM) images of AIS demonstrated abnormal osteocytes that were more rounded and cobblestone-like in shape and were aligned in irregular clusters with shorter and disorganized canaliculi. Further quantitative analysis with FITC-Imaris technique showed a significant reduction in the canalicular number and length as well as an increase in lacunar volume and area in AIS. SEM with energy-dispersive X-ray spectroscopy analysis demonstrated a lower calcium-to-phosphorus ratio at the perilacunar/canalicular region. Moreover, microindentaion results revealed lower values of Vickers hardness and elastic modulus in AIS when compared with controls. In addition, in the parallel study of 99 AIS (27 with severe Cobb angle of 65.8 ± 14.1° and 72 with mild Cobb angle of 26.6 ± 9.1°) with different curve severity, the serum osteocalcin level was found to be significantly and negatively associated with the Cobb angle. In summary, the findings in this series of studies demonstrated the potential link of abnormal osteocyte lacuno-canalicular network structure and function to the observed abnormal bone mineralization in AIS, which may shed light on etiopathogenesis of AIS.-Chen, H., Zhang, J., Wang, Y., Cheuk, K.-Y., Hung, A. L. H., Lam, T.-P., Qiu, Y., Feng, J. Q., Lee, W. Y. W., Cheng, J. C. Y. Abnormal lacuno-canalicular network and negative correlation between serum osteocalcin and Cobb angle indicate abnormal osteocyte function in adolescent idiopathic scoliosis.


A population of CD4hiCD38hi T cells correlates with disease severity in patients with acute malaria.
CD4+ T cells are critical mediators of immunity to Plasmodium spp. infection, but their characteristics during malarial episodes and immunopathology in naturally infected adults are poorly defined. Flow cytometric analysis of PBMCs from patients with either P. falciparum or P. knowlesi malaria revealed a pronounced population of CD4+ T cells co-expressing very high levels of CD4 and CD38 we have termed CD4hiCD38hi T cells. We set out to gain insight into the function of these novel cells. CD4+ T cells from 18 patients with P. falciparum or P. knowlesi malaria were assessed by flow cytometry and sorted into populations of CD4hiCD38hi or CD4norm T cells. Gene expression in the sorted populations was assessed by qPCR and NanoString. CD4hiCD38hi T cells expressed high levels of CD4 mRNA and canonical type 1 regulatory T-cell (TR1) genes including IL10, IFNG, LAG3 and HAVCR2 (TIM3), and other genes with relevance to cell migration and immunomodulation. These cells increased in proportion to malaria disease severity and were absent after parasite clearance with antimalarials. In naturally infected adults with acute malaria, a prominent population of type 1 regulatory T cells arises that can be defined by high co-expression of CD4 and CD38 (CD4hiCD38hi) and that correlates with disease severity in patients with falciparum malaria. This study provides fundamental insights into T-cell biology, including the first evidence that CD4 expression is modulated at the mRNA level. These findings have important implications for understanding the balance between immunity and immunopathology during malaria.


Effect of lactation counselling on subclinical mastitis among Bangladeshi women.
Subclinical mastitis, defined as raised milk sodium/potassium (Na/K) in the absence of clinical symptoms, is associated with poor infant weight gain and increased risk of mother-to-child HIV transmission. Similarly to clinical mastitis, subclinical mastitis appears to have multiple causes, one being poor lactation practice. We investigated whether a single session of lactation counselling around the time of delivery could decrease subclinical mastitis among women in rural Bangladesh. The counselling messages included the importance of giving colostrum, exclusive breastfeeding, feeding on demand and how to achieve good positioning and attachment. Women who had delivered prior to our instituting the counselling intervention (mean 1 month) constituted the uncounselled group. At 1 month postpartum, sodium and potassium were measured in spot milk samples from each breast by flame photometry and interleukin-8 (IL8) by ELISA. Geometric mean Na/K ratios and the proportion of milk samples with raised Na/K ratio were significantly (p < 0.05) lower in the counselled group [mean 0.42, 95% confidence interval (CI) 0.39-0.45, n = 116 samples; 10% Na/K ratio > 0.6] than in the non-counselled group [mean 0.49, 95% CI 0.45-0.53, n = 127; 25% Na/K ratio > 0.6]. Milk IL8 concentration did not differ between groups. The results show that among Bangladeshi women a simple lactation counselling intervention could improve lactation practice in ways likely to improve infant health.


Using historical biogeography to test for community saturation.
Saturation is the idea that a community is effectively filled with species, such that no more can be added without extinctions. This concept has important implications for many areas of ecology, such as species richness, community assembly, invasive species and climate change. Here, we illustrate how biogeography can be used to test for community saturation, when combined with data on local species richness, phylogeny and climate. We focus on a clade of frogs (Terrarana) and the impact of the Great American Biotic Interchange on patterns of local richness in Lower Middle America and adjacent regions. We analyse data on species richness at 83 sites and a time-calibrated phylogeny for 363 species. We find no evidence for saturation, and show instead that biotic interchange dramatically increased local richness in the region. We suggest that historical biogeography offers thousands of similar long-term natural experiments that can be used to test for saturation.


Legislative provisions for standardised tobacco packaging and changes in public support in 27 European countries.
Global adoption of standardised packaging requirements for tobacco products is a victory for public health, but their proliferation and impacts rely partly on public support. How this is related to legislation remains underassessed. This study explored change over time in public support for standardised packaging in countries with varying degrees of legislative provisions. We used data from 27 European countries, collected from 2017 (n=28, 300) and 2020 (n=27, 901) waves of the Eurobarometer survey, to assess self-reported support for standardised packaging regulations among both smokers and non-smokers. Countries were grouped into three categories of policy adoption (policy implemented; policy legislated; no legislation) and changes in support were assessed using multilevel Poisson regression models. In 2020, public support for standardised packaging was 71% (95% CI 68% to 74%) in countries that implemented standardised packaging legislation, 57% (55% to 60%) in countries that had legislated but not yet implemented legislation and 41% (40%to 42%) in countries with no legislation. Compared with 2017, this represented a relative change of +8% (1% to 15%), +12% (5% to 21%) and -5% (95% CI -2% to -8%), respectively, in the three country categories. Among smokers, there was no indication of change in support across the three groups. Among non-smokers, support increased in countries with existing legislation (adjusted prevalence ratio [aPR]=1.14, 95% CI 1.06 to 1.23) and decreased in countries with no legislation (aPR=0.93, 0.90 to 0.97). Public support for standardised packaging regulations increased in countries implementing and legislating for these measures, particularly among non-smokers. An overall increase in support provides reassurance for policymakers defending policy action on tobacco packaging, as well as for those seeking to implement standardised packaging in their own countries. .


The importance of imprinting in the human placenta.
As a field of study, genomic imprinting has grown rapidly in the last 20 years, with a growing figure of around 100 imprinted genes known in the mouse and approximately 50 in the human. The imprinted expression of genes may be transient and highly tissue-specific, and there are potentially hundreds of other, as yet undiscovered, imprinted transcripts. The placenta is notable amongst mammalian organs for its high and prolific expression of imprinted genes. This review discusses the development of the human placenta and focuses on the function of imprinting in this organ. Imprinting is potentially a mechanism to balance parental resource allocation and it plays an important role in growth. The placenta, as the interface between mother and fetus, is central to prenatal growth control. The expression of genes subject to parental allelic expression bias has, over the years, been shown to be essential for the normal development and physiology of the placenta. In this review we also discuss the significance of genes that lack conservation of imprinting between mice and humans, genes whose imprinted expression is often placental-specific. Finally, we illustrate the importance of imprinting in the postnatal human in terms of several human imprinting disorders, with consideration of the brain as a key organ for imprinted gene expression after birth.


Human immunity against EBV-lessons from the clinic.
The mammalian immune system has evolved over many millennia to be best equipped to protect the host from pathogen infection. In many cases, host and pathogen have coevolved, each acquiring sophisticated ways of inducing or protecting from disease. Epstein-Barr virus (EBV) is a human herpes virus that infects >90% of individuals. Despite its ubiquity, infection by EBV is often subclinical; this invariably reflects the necessity of the virus to preserve its host, balanced with sophisticated host immune mechanisms that maintain viral latency. However, EBV infection can result in various, and often fatal, clinical sequelae, including fulminant infectious mononucleosis, hemophagocytic lymphohistiocytosis, lymphoproliferative disease, organomegaly, and/or malignancy. Such clinical outcomes are typically observed in immunosuppressed individuals, with the most extreme cases being Mendelian primary immunodeficiencies (PIDs). Although these conditions are rare, they have provided critical insight into the cellular, biochemical, and molecular requirements for robust and long-lasting immunity against EBV infection. Here, we review the virology of EBV, mechanisms underlying disease pathogenesis in PIDs, and developments in immune cell-mediated therapy to treat disorders associated with or induced by EBV infection.


Thiamine outflow from the enterocyte: a study using basolateral membrane vesicles from rat small intestine.
1. Rat small intestinal basolateral membrane vesicles (BLMVs) were prepared and found to be 31% non-vesiculated and 69% vesiculated, 4.9% right side out and 63.8% inside out. 2. Thiamine uptake by BLMVs followed a hyperbolic time course reaching equilibrium after 60-90 min incubation. Uptake was not affected by the transmembrane potential or by the presence or absence of Na+ or K+ in the incubation medium. 3. At concentrations below 1.25 microM, [3H]thiamine was taken up mainly by a saturable mechanism with an apparent Michaelis-Menten constant (Km) = 1.32 microM and maximal flux (Jmax) = 1.93 pmol (mg protein)-1 (4 s)-1. At higher concentrations, a non-saturable mechanism prevailed. 4. Only 29% of [3H]thiamine taken up by the vesicles was membrane bound, the remaining being translocated into the vesicular space. No thiamine phosphoesters could be detected inside the vesicles. 5. In the absence of ATP, the Na(+)-K(+)-ATPase inhibitors ouabain, frusemide and vanadate reduced thiamine uptake by 35, 30 and 15% respectively. 6. In experiments conducted with K+ inside the vesicles and Na+, Mg2+ and ATP outside, the time course of thiamine uptake by BLMVs displayed an overshoot (80-90% increment) at 30 s incubation as compared to controls. When ATP was replaced with phosphocreatine, or when NaCl was replaced with isosmotic amounts of KCl, the overshoot disappeared. 7. The thiamine analogues pyrithiamine, amprolium and 4'-oxythiamine decreased the ATPase-dependent transport of [3H]thiamine by 100, 86 and 31% respectively. 8. These results provide evidence that the transport of thiamine by BLMVs is coupled directly to the hydrolysis of ATP (primary active transport).


Divergent impact of progesterone and medroxyprogesterone acetate (Provera) on nuclear mitogen-activated protein kinase signaling.
The impact of progestins on estrogen-inducible mechanisms of neuroprotection was investigated. Previously, we showed that estrogen and progesterone are neuroprotective against excitotoxicity, whereas the synthetic progestin medroxyprogesterone acetate (MPA; Provera) is not. Here, we demonstrate that 17beta-estradiol (E2) and progesterone (P4) treatment of hippocampal neurons attenuated the excitotoxic glutamate-induced rise in intracellular calcium concentration. Although MPA had no effect alone, MPA completely antagonized E2-induced attenuation of intracellular calcium concentration. Activation of extracellular receptor kinase (ERK) is required for estrogen-induced neuroprotection and calcium regulation. Paradoxically, E2, P4, and MPA all elicited similar rapid and transient activation of ERK, presenting a contradiction between the dependence on ERK for gonadal hormone-induced neuroprotection and the lack of neuroprotection induced by MPA. Subcellular analysis of ERK demonstrated that the phospho-ERK signal is transduced to the nucleus only by E2 and P4, not by MPA. These results indicate that the profile of nuclear translocation of ERK is consistent with the neuroprotective profile. Further, the E2-induced nuclear translocation of ERK was blocked by coadministration of MPA. Results of this study reveal that nuclear ERK induction by ovarian steroids is predictive of the neuroprotective effects of estrogen and progestin treatments, revealing a hitherto unrecognized divergence of progestin signaling through the src/MAPK pathway. These results have much broader implications encompassing the impact of progestins on estrogen-mediated effects in multiple tissues. The recent results from the Women's Health Initiative trial, which used MPA as the progestinal agent, indicate that differences between progestin formulations are crucial to health outcomes in women.


Outlier detection and handling for robust 3-D active shape models search.
This paper presents a new outlier handling method for volumetric segmentation with three-dimensional (3-D) active shape models. The method is based on a shape metric that is invariant to scaling, rotation and translation by using the ratio of interlandmark distances as a local shape dissimilarity measure. Tolerance intervals for the descriptors are calculated from the training samples and used as a statistical tolerance model to infer the validity of the feature points. A replacement point is then suggested for each outlier based on the tolerance model and the position of the valid points. A geometrically weighted fitness measure is introduced for feature point detection, which limits the presence of outliers and improves the convergence of the proposed segmentation framework. The algorithm is immune to the extremity of the outliers and can handle a highly significant presence of erroneous feature points. The practical value of the technique is validated with 3-D magnetic resonance (MR) segmentation tasks of the carotid artery and myocardial borders of the left ventricle.


New iron pyridylamino-bis(phenolate) catalyst for converting CO2 into cyclic carbonates and cross-linked polycarbonates.
The atom-efficient reaction of CO2 with a variety of epoxides has been efficiently achieved employing iron pyridylamino-bis(phenolate) complexes as bifunctional catalysts. The addition of a Lewis base co-catalyst allowed significant reduction in the amount of iron complex needed to achieve high epoxide conversions. The possibility of controlling the selectivity of the reaction towards either cyclic carbonate or polycarbonate was evaluated. An efficient switch in selectivity could be achieved when cyclic epoxides such as cyclohexene oxide and the seldom explored 1,2-epoxy-4-vinylcyclohexane were used as substrates. The obtained poly(vinylcyclohexene carbonate) presents pending vinyl groups, which allowed post-synthetic cross-linking by reaction with 1,3-propanedithiol. The cross-linked polycarbonate displayed a substantial increase in the glass transition temperature and chemical resistance, thus opening new opportunities for the application of these green polymers.


Synthetic studies towards putative yuremamine using an iterative C(sp(3))-H arylation strategy.
An overview of an iterative, 8-aminoquinoline (AQ)-directed C(sp(3))-H arylation strategy towards the pyrroloindole structure initially assigned to the alkaloid yuremamine is described. During initial efforts using a model indane system, it was discovered that the iodoresorcinol unit was not a viable C(sp(3))-H arylation partner when masked as its dimethyl ether but upon switching to a MOM group, the ether oxygen served to stabilise the high valent Pd intermediate during the reaction, thus promoting reductive elimination and leading to acceptable yields of the C(sp(3))-H arylation product. The second C(sp(3))-H arylation with an iodopyrogallol gave a 1,3-diarylated model yuremamine system possessing the desired 1,3-cis relationship. When the successful model studies were applied to a pyrroloindole system in pursuit of yuremamine, it became apparent that C9 underwent competing C(sp(2))-H arylation if left vacant, but installing a tryptamine side chain at this site prevented the desired C(sp(3))-H arylation from occurring altogether. However, a C9-methyl pyrroloindole underwent iterative C(sp(3))-H arylation at C1 with an iodoresorcinol followed by C3 with an iodopyrogallol to give a diarylated product with the aryl groups in the undesired 1,3-trans-relationship, arising from epimerisation at C1 during the second C(sp(3))-H arylation event. Although the synthesis of putative yuremamine was not accomplished, several findings are disclosed that will serve as useful additions to the burgeoning field of directed C(sp(3))-H arylations and related C-H functionalization reactions.


Clay improvement with burned olive waste ash.
Olive oil is concentrated in the Mediterranean basin countries. Since the olive oil industries are incriminated for a high quantity of pollution, it has become imperative to solve this problem by developing optimized systems for the treatment of olive oil wastes. This study proposes a solution to the problem. Burned olive waste ash is evaluated for using it as clay stabilizer. In a laboratory, bentonite clay is used to improve olive waste ash. Before the laboratory, the olive waste is burned at 550°C in the high temperature oven. The burned olive waste ash was added to bentonite clay with increasing 1% by weight from 1% to 10%. The study consisted of the following tests on samples treated with burned olive waste ash: Atterberg Limits, Standard Proctor Density, and Unconfined Compressive Strength Tests. The test results show promise for this material to be used as stabilizer and to solve many of the problems associated with its accumulation.


Application of the Goutallier/Fuchs Rotator Cuff Classification to the Evaluation of Hip Abductor Tendon Tears and the Clinical Correlation With Outcome After Repair.
To assess the reliability and reproducibility of the Goutallier/Fuchs classification for the evaluation of abductor tendon tears of the hip, as well as to identify the relation between preoperative tear size, abductor muscle quality, and the success of endoscopic tendon repair. This is a retrospective review of 30 consecutive endoscopic abductor tendon repairs performed by a single surgeon over a 2-year period. Preoperative magnetic resonance imaging scans were reviewed, and the muscle was assigned a grade according to the Goutallier/Fuchs classification. Patient-rated outcome scores--visual analog scale score, Hip Outcome Score (HOS), and modified Harris Hip Score (mHHS)--were collected preoperatively and at a minimum of 2 years postoperatively. Intraobserver and interobserver reliability for muscle grading was calculated. Postoperative outcome measures were compared with preoperative tear size, muscle grade, and repair type to assess for correlations. Of the 30 hips included in the study, over 75% were classified as grade 1 (n = 15) or grade 2 (n = 8). The intraobserver reliability and interobserver reliability of the classification system averaged 0.872 and 0.916, respectively. Two patients (grades 3 and 4) had repair failure and underwent muscle transfer. In the remaining 28 hips, improvement was seen in the visual analog scale score (6.0 v 1.7, P < .0001), HOS-Activities of Daily Living subscale score (58.8 v 83.4, P < .0001), HOS-Sport-Specific subscale score (40.0 v 75.0, P < .0001), and mHHS (55.6 points v 81.1 points, P < .0001) postoperatively. Increasing preoperative fatty infiltration and atrophy correlated with increased postoperative pain levels (regression coefficient, 0.93; P < .001) and decreased postoperative HOS-Activities of Daily Living subscale scores (regression coefficient, -3.36; P = .011), HOS-Sport-Specific subscale scores (regression coefficient, -5.63; P = .016), mHHS values (regression coefficient, -3.50; P = .0008), and patient satisfaction (regression coefficient, -1.04; P < .0001). Patient age, tear size, or repair type (double v single row) did not affect postoperative outcomes. The Goutallier/Fuchs classification system can be reliably and reproducibly applied to the evaluation of abductor tendon tears of the hip and appears to correlate with patient-rated outcomes after repair. Increasing preoperative muscle fatty atrophy correlates with increased patient pain and decreased patient satisfaction and functional outcomes after repair. Level IV, prognostic case series.


The imagework method in health and social science research.
Existing alongside the traditional forms of qualitative social science research, there is a set of potential research methods that derive from experiential groupwork and the humanistic human potential movement and are only slightly used by researchers. Social science research has barely begun to use these powerful strategies that were developed originally for personal and group change but that are potentially applicable to the research domain. This article will locate these methods within the qualitative research domain and propose a novel view of their value. The study of the actual and potential use of one of these methods, imagework, will be the particular focus of this article. References to the use of artwork, sculpting, psychodrama, gestalt, and dreamwork will also be made. The hypothesis underpinning the author's approach is that experiential research methods such as imagework can elicit implicit knowledge and self-identifies of respondents in a way that other methods cannot.


Metal doped nanosized titania used for the photocatalytic degradation of rhodamine B dye under visible-light.
Metal-doped anatase nanosized titania photocatalysts were successfully synthesized using a sol-gel process. Different amounts of the dopants (0.2, 0.4, 0.6, 0.8 and 1.0%) of the metals (Ag, Ni, Co and Pd) were utilized. The UV-Vis spectra (solid state diffuse reflectance spectra) of the doped nanoparticles exhibited a red shift in the absorption edge as a result of metal doping. The metal-doped nanoparticles were investigated for their photocatalytic activity under visible-light irradiation using Rhodamine B (Rh B) as a control pollutant. The results obtained indicate that the metal-doped titania had the highest activity at 0.4% metal loading. The kinetic models revealed that the photodegradation of Rh B followed a pseudo first order reaction. From ion chromatography (IC) analysis the degradation by-products Rhodamine B fragments were found to be acetate, chloride, nitrite, carbonate and nitrate ions.


Inhibition of glioma progression by a newly discovered CD38 inhibitor.
Glioma, the most common cancer of the central nervous system, has very poor prognosis and no effective treatment. It has been shown that activated microglia/macrophages in the glioma tumor microenvironment support progression. Hence, inhibition of the supporting effect of these cells may constitute a useful therapeutic approach. Recently, using a syngeneic mouse glioma progression model, we showed that the ectoenzyme CD38 regulated microglia activation and, in addition, that the loss of CD38 from the tumor microenvironment attenuated glioma progression and prolonged the life span of the tumor-bearing mice. These studies, which employed wild-type (WT) and Cd38(-/-) C57BL/6J mice, suggest that inhibition of CD38 in glioma microenvironment may be used as a new therapeutic approach to treat glioma. Our study tested this hypothesis. Initially, we found that the natural anthranoid, 4,5-dihydroxyanthraquinone-2-carboxylic acid (rhein), and its highly water-soluble tri-potassium salt form (K-rhein) are inhibitors of CD38 enzymatic (nicotinamide adenine dinucleotide glycohydrolase) activity (IC50 = 1.24 and 0.84 μM, respectively, for recombinant mouse CD38). Treatment of WT, but not Cd38(-/-) microglia with rhein and K-rhein inhibited microglia activation features known to be regulated by CD38 (lipopolysaccharide/IFN-γ-induced activation, induced cell death and NO production). Furthermore, nasal administration of K-rhein into WT, but not Cd38(-/-) C57BL/6J, mice intracranially injected with GL261 cells substantially and significantly inhibited glioma progression. Hence, these results serve as a proof of concept, demonstrating that targeting CD38 at the tumor microenvironment by small-molecule inhibitors of CD38, for example, K-rhein, may serve as a useful therapeutic approach to treat glioma.


Electrochemical and theoretical investigations of the role of the appended base on the reduction of protons by [Fe2(CO)4(κ2-PNP(R)(μ-S(CH2)3S] (PNP(R) ={Ph2PCH2}2NR, R=Me, Ph).
The behavior of [Fe(2)(CO)(4)(κ(2)-PNP(R))(μ-pdt)] (PNP(R) =(Ph(2)PCH(2))(2)NR, R=Me (1), Ph (2); pdt=S(CH(2))(3)S) in the presence of acids is investigated experimentally and theoretically (using density functional theory) in order to determine the mechanisms of the proton reduction steps supported by these complexes, and to assess the role of the PNP(R) appended base in these processes for different redox states of the metal centers. The nature of the R substituent of the nitrogen base does not substantially affect the course of the protonation of the neutral complex by CF(3)SO(3)H or CH(3)SO(3)H; the cation with a bridging hydride ligand, 1 μH(+) (R=Me) or 2 μH(+) (R=Ph) is obtained rapidly. Only 1 μH(+) can be protonated at the nitrogen atom of the PNP chelate by HBF(4)·Et(2)O or CF(3)SO(3)H, which results in a positive shift of the proton reduction by approximately 0.15 V. The theoretical study demonstrates that in this process, dihydrogen can be released from a η(2)-H(2) species in the Fe(I)Fe(II) state. When R=Ph, the bridging hydride cation 2 μH(+) cannot be protonated at the amine function by HBF(4)·Et(2)O or CF(3)SO(3)H, and protonation at the N atom of the one-electron reduced analogue is also less favored than that of a S atom of the partially de-coordinated dithiolate bridge. In this situation, proton reduction occurs at the potential of the bridging hydride cation, 2 μH(+). The rate constants of the overall proton reduction processes are small for both complexes 1 and 2 (k(obs) ≈4-7 s(-1)) because of the slow intramolecular proton migration and H(2) release steps identified by the theoretical study.


Prevalence of Bladder and Bowel Dysfunction in Duchenne Muscular Dystrophy Using the Childhood Bladder and Bowel Dysfunction Questionnaire.
Lower urinary tract symptoms (LUTS) and gastrointestinal (GI) problems are common in Duchenne muscular dystrophy (DMD), but not systematically assessed in regular care. We aimed to determine the prevalence of bladder and bowel dysfunction (BBD) in DMD patients compared with healthy controls (HC). The Childhood Bladder and Bowel Dysfunction Questionnaire (CBBDQ) based on the International Rome III criteria and the International Children's Continence Society was filled out by 57 DMD patients and 56 HC. Additionally, possible associations of BBD with, for example, medication use or quality of life were evaluated in an additional questionnaire developed by experts. In 74% of patients versus 56% of HC ≥ 1 LUTS (n.s.) were reported, 68% of patients versus 39% of HC reported ≥1 bowel symptom (p = 0.002) and 53% of patients versus 30% of HC reported combined LUTS and bowel symptoms (p = 0.019). A negative impact of BBD on daily life functioning was reported by 42% of patients. These data underscore that standard screening for BBD is needed and that the CBBDQ could be of added value to optimize DMD care.


Potential impact of increased use of biocides in consumer products on prevalence of antibiotic resistance.
There has recently been much controversy surrounding the increased use of antibacterial substances in a wide range of consumer products and the possibility that, as with antibiotics, indiscriminate use of biocides might contribute to the overall pattern of susceptibility in the general environment and in the clinic. Such speculation, based on the isolation of resistant mutants from in vitro monoculture experiments, is not reflected by an emergence of biocide-resistant strains in vivo. This review provides a broad coverage of the biocide and resistance literature and evaluates the potential risks, perceived from such laboratory monoculture experiments, against evidence gathered over 50 years of field studies. An explanation for the continued effectiveness of broad-spectrum biocidal agents against the decline in efficacy of therapeutic agents is provided based on the fitness costs of resistance and the ubiquity of naturally occurring substances that possess antibacterial effect. While we conclude from this review of the literature that the incorporation of antibacterial agents into a widening sphere of personal products has had little or no impact on the patterns of microbial susceptibility observed in the environment, the associated risks remain finite. The use of such products should therefore be associated with a clear demonstration of added value either to consumer health or to the product life. Hygienic products should therefore be targeted to applications for which the risks have been established.


Variants of BALB/c 3T3 cells lacking complex gangliosides retain a fibronectin matrix and spread normally on fibronectin-coated substrates.
Evidence has accumulated that di- and trisialogangliosides are involved in the interaction of cells with fibronectin. We have therefore tested the ability of variants of BALB/c 3T3 deficient in such gangliosides to organize a fibronectin matrix and to spread on fibronectin-coated substrates. Whereas BALB/c 3T3 cells contained gangliosides GM3, GM1, and GD1a, direct chemical analysis showed that five out of six variants isolated contained no detectable GD1a. By the overlaying of thin layer chromatograms of cellular gangliosides with 125I-cholera toxin, these variants were also found to lack ganglioside GM1. In contrast, the sialogalactoprotein profile of these cells, analyzed using an 125I-ricin/SDS polyacrylamide gel overlay technique, was similar to that of the parent cell line. All variants organized an extensive fibronectin matrix comparable to that of BALB/c 3T3, as shown using either immunofluorescence or lactoperoxidase-catalyzed iodination. The variants could also spread on fibronectin-coated substrates and adopt a morphology similar to that of BALB/c 3T3 cells, with little or no difference in the concentration of fibronectin required for 50% cell spreading. Cell spreading of the variants was accompanied by the formation of focal contacts and microfilament bundles, in a manner closely resembling that seen with BALB/c 3T3 cells. Treatment of BALB/c 3T3 cells with neuraminidase, which converts much of the cellular GD1a to GM1, did not affect cell spreading on fibronectin. The results clearly demonstrate that complex gangliosides are not essential for retention of a fibronectin matrix or for spreading on fibronectin-coated substrates.


A prognostic model for advanced stage nonsmall cell lung cancer. Pooled analysis of North Central Cancer Treatment Group trials.
A pooled analysis was performed to examine the impact of pretreatment factors on overall survival (OS) and time to progression (TTP) in patients with advanced-stage nonsmall cell lung cancer (NSCLC) and to construct a prediction equation for OS using pretreatment factors. A pooled data set of 1053 patients from 9 North Central Cancer Treatment Group trials was used. Age, gender, Eastern Cooperative Oncology Group performance status (PS), tumor stage (Stage IIIB vs. Stage IV), body mass index (BMI), creatinine level, hemoglobin (Hgb) level, white blood cell (WBC) count, and platelet count were evaluated for their prognostic significance in both univariate and multivariate analyses by using a Cox proportional-hazards model. Patients who had high WBC counts, low Hgb levels, PS >0, BMI < 18.5 kg/m2, and TNM Stage IV disease had significantly worse TTP and OS. Patients who had Stage IV disease with a high WBC count had a particularly poor prognosis. An equation to predict the OS of patients with Stage IV NSCLC based on pretreatment PS, BMI, Hgb level, and WBC count was constructed. In addition to the widely accepted prognostic factors of PS, BMI, and disease stage, both of the readily available laboratory parameters of Hgb level and WBC count were found to be significant prognostic factors for OS and TTP in patients with advanced-stage NSCLC. The authors' prediction equation can be used to evaluate the benefit of a treatment in Phase II trials by comparing the observed survival of a cohort with its expected survival by using the patients' own prognostic factors in place of comparisons with historic data that may have substantially different baseline patient characteristics.


Adjusting hemodialysis dose for protein catabolic rate.
Patients dialyzed with equal eKt/V may have huge variations in their urea concentrations. Urea generation rate, distribution volume and renal clearance were determined in 205 hemodialysis sessions of 33 patients with double pool urea kinetic modeling using dialyzer clearance from online monitoring. From these data, optimized prescriptions were computed. In simulated dialysis sessions, the HEMO standard-dose equivalent clearance was not sufficient to keep time-averaged concentration (TAC) and average predialysis concentration (PAC) of urea below the defined upper limits (20 and 30 mmol/l), if normalized protein catabolic rate (nPCR) was greater than 1.3 g/kg/day. Protein catabolic rate was taken into account in the optimized prescription by cutting high urea concentrations. If patients having high urea concentrations with conventional clearance will benefit from higher dialysis dose - an unconfirmed hypothesis - this approach helps in identifying those who need more than three sessions per week.


Optimization of Transesterification Reactions with CLEA-Immobilized Feruloyl Esterases from Thermothelomyces thermophila and Talaromyces wortmannii.
Feruloyl esterases (FAEs, E.C. 126.96.36.199) are biotechnologically important enzymes with several applications in ferulic acid production from biomass, but also in synthesis of hydroxycinnamic acid derivatives. The use of such biocatalysts in commercial processes can become feasible by their immobilization, providing the advantages of isolation and recycling. In this work, eight feruloyl esterases, immobilized in cross-linked enzyme aggregates (CLEAs) were tested in regard to their transesterification performance, towards the production of prenyl ferulate (PFA) and arabinose ferulate (AFA). After solvent screening, comparison with the activity of respective soluble enzymes, and operational stability tests, FAE125 was selected as the most promising biocatalyst. A central composite design revealed the optimum conditions for each transesterification product, in terms of water content, time, and substrate ratio for both products, and temperature and enzyme load additionally for prenyl ferulate. The optimum product yields obtained were 83.7% for PFA and 58.1% for AFA. FAE125 CLEAs are stable in the optimum conditions of transesterification reactions, maintaining 70% residual activity after five consecutive reactions. Overall, FAE125 CLEAs seem to be able to perform as a robust biocatalyst, offering satisfactory yields and stability, and thus showing significant potential for industrial applications.


Male sexual function and pelvic floor surgery of their female partner: A one-year follow-up study.
The aim of this study was to examine if sexual function of male partners changed after surgery for pelvic floor disorders and to explore associated factors. This was an observational follow-up study at the Gynecological Department at the St Olavs University Hospital, Trondheim. The sample consisted of 35 male partners of women scheduled for pelvic organ prolapse or stress urinary incontinence surgery. Self-administered questionnaires, containing validated instruments as well as exploratory questions, were sent to women and their partners before and one year after pelvic floor surgery. Vaginal dimensions were measured in all women according to the Pelvic Organ Prolapse Quantification System, both before and after the surgery. The Brief Sexual Function Instrument and the presence of erectile dysfunction. One year after pelvic floor surgery, scores for sexual drive, erection and overall satisfaction from the Brief Sexual Function Instrument were unchanged; the ejaculation score (range 0-4) had mildly improved from a range of 4 (median 4) to a range of 3.5 (median 4), (p = 0.014). The proportion of men with erectile dysfunction was unchanged, while the proportion of men reporting vaginal wind had significantly decreased (p = 0.016). None of the baseline factors, subjective experiences or vaginal dimensions at baseline or follow-up were associated with the improved ejaculation score; only a reduction in the proportion of men reporting their partners with dyspareunia (ns) was significantly correlated (Spearman's rho 0.42, p = 0.019). Sexual function of male partners was unchanged or mildly improved after pelvic floor surgery.


The Suicidal Behaviors Questionnaire-Revised (SBQ-R) and its Chinese version (C-SBQ-R): Further validity testing using the culture, comprehension, and translation bias procedure.
The Suicidal Behaviors Questionnaire-Revised (SBQ-R), a 4-item instrument for assessing and screening suicide-related thoughts and behaviors, has been translated into several different languages. In this study, we examined the psychometric properties of the SBQ-R and a new Chinese version of the instrument (C-SBQ-R) for college student samples. We used the culture, comprehension, and translation bias (CCT) procedure to examine the responses of independent Chinese college student samples who completed the C-SBQ-R (N₁ = 298) and the SBQ-R (N₂ = 292) and a U.S. college student sample (N₃ = 320) who completed the SBQ-R. We found that the samples' C-SBQ-R and SBQ-R scale scores showed acceptable internal consistency reliability estimates, factor structures, and concurrent validity estimates. Measurement invariance was established across groups differing in (a) language but not culture, (b) culture but not language, and (c) both language and culture. Receiver operating characteristic curve analysis results showed that the cutoff score of 7 was similar for both the C-SBQ-R and the SBQ-R in screening for suicide-related thoughts and behaviors. Our findings provided empirical support that the psychometric properties of the C-SBQ-R and the SBQ-R are similar; both could be used to assess the suicide-related thoughts and behaviors constructs. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Quantitative assessment of p-glycoprotein expression and function using confocal image analysis.
P-glycoprotein is implicated in clinical drug resistance; thus, rapid quantitative analysis of its expression and activity is of paramout importance to the design and success of novel therapeutics. The scope for the application of quantitative imaging and image analysis tools in this field is reported here at "proof of concept" level. P-glycoprotein expression was utilized as a model for quantitative immunofluorescence and subsequent spatial intensity distribution analysis (SpIDA). Following expression studies, p-glycoprotein inhibition as a function of verapamil concentration was assessed in two cell lines using live cell imaging of intracellular Calcein retention and a routine monolayer fluorescence assay. Intercellular and sub-cellular distributions in the expression of the p-glycoprotein transporter between parent and MDR1-transfected Madin-Derby Canine Kidney cell lines were examined. We have demonstrated that quantitative imaging can provide dose-response parameters while permitting direct microscopic analysis of intracellular fluorophore distributions in live and fixed samples. Analysis with SpIDA offers the ability to detect heterogeniety in the distribution of labeled species, and in conjunction with live cell imaging and immunofluorescence staining may be applied to the determination of pharmacological parameters or analysis of biopsies providing a rapid prognostic tool.


The results of cholecystectomy at a district general hospital. A reappraisal of operative cholangiography.
A total of 487 cholecystectomies were performed at one hospital over a 44-month period by surgeons who differed considerably in the frequency with which they performed operative cholangiography. There were no differences in the frequency or severity of postoperative complications or recurrent symptoms between patients who did and those who did not undergo operative cholangiography; nor were any differences demonstrable between patients operated on by different surgeons. Although it is established that operative cholangiography will demonstrate otherwise unsuspected common bile duct stones in some patients, it has not yet been clearly proved that these stones would cause later complications if left undisturbed. The clinical advantages of operative cholangiography are not sufficiently clear to preclude their assessment by prospective controlled studies involving large numbers of patients.


Possible correlations between the deficient affective experience and negative symptoms of psychosis: an exploratory study in (forensic) psychiatric patients.
Deficient affective experience (DAE), the affective and perhaps fundamental component of psychopathy, has some resemblance to the core affective disturbances of schizophrenia. There has, however, been little attempt to test relationships between these concepts, but this could be useful because of the high prevalence of schizophrenia among people in forensic mental health services and evidence that the DAE may be a useful predictor of violent behaviour. Our primary aim was to explore possible correlations between DAE and negative symptoms of schizophrenia and to test the hypotheses that the DAE would differentiate people with and without personality disorder co-morbidity in a sample with psychosis and, separately, people with main diagnoses of personality disorder or schizophrenia. On the basis of an interview and a review of institutional files, DAE total and facet scores were calculated and compared with scores on the negative symptom scale of the Positive and Negative Syndrome Scale among forensic and general psychiatric patients. Partial correlation and rank order coefficients were calculated. We found no correlation between total DAE scores and total negative Positive and Negative Syndrome Scale scores among patients with psychosis, suggesting that the concepts probably do not overlap. Our hypotheses on diagnostic associations with the DAE were partially sustained. DAE scores differentiated psychosis and personality disorder groups but did not differentiate psychosis groups with and without personality pathology. This study, using typical clinical samples, suggested that one aspect of the affective disturbance of schizophrenia has little in common with the deficient emotional experience of psychopath. Measurement of DAE is unlikely to help detect co-morbid personality disorder among people with psychosis, but given our small sample size and the rarity of study in this field, further research would be warranted, perhaps also including a measure of incongruity of affect.


Sex differences in stress reactivity of hippocampal BDNF in mice are associated with the female preponderance of decreased locomotor activity in response to restraint stress.
The incidence and prevalence of depression is higher in women than in men, but the cause of this sex discrepancy remains unknown. Brain-derived neurotrophic factor (BDNF) is a key protein for maintaining neuronal integrity. The purpose of this study was to investigate the female preponderance in behavioral responsivity to restraint stress focusing on the stress reactivity of BDNF in the hippocampus. Male and female ICR mice were exposed to a 3-h session of restraint stress. Plasma corticosterone was measured by high-performance liquid chromatography. BDNF mRNA expression in the whole hippocampus was measured by quantitative real-time reverse transcription-polymerase chain reaction. Wheel-running activity was monitored during the dark period. In response to restraint stress, the increase in levels of serum corticosterone was higher in female than in male mice. Restraint stress resulted in decreased voluntary wheel-running behavior that was greater in female than male animals. In addition to these sex differences in stress reactivity, we found a significant sex difference in BDNF levels in the hippocampus of restraint-stressed mice; total BDNF levels significantly decreased in female mice, but not in male mice in response to the stress. Furthermore, BDNF exon I and IV mRNA expression also showed the same tendency. These data indicate that the reduction in levels of voluntary wheel-running activity in response to stress can be significantly influenced by sex. Moreover, our findings suggest a link between the sex differences in this behavioral response to stress and differential stress reactivity in the production of BDNF in the hippocampus.


Accounting for apparent "reverse" racial disparities in Department of Veterans Affairs (VA)-based medical care: influence of out-of-VA care.
Conclusions regarding racial differences in care following a newly elevated prostate-specific antigen (PSA) test at the Department of Veterans Affairs (VA) may differ depending on whether follow-up care outside the VA is considered. Consecutive Philadelphia, Pa, VA patients with newly elevated PSA tests (n = 183) were interviewed 1 year after baseline. Among exclusive VA users, Blacks had higher rates of urology referrals and prostate biopsies compared with Whites. However, these racial differences were attenuated when care obtained outside the VA also was considered.


Enhanced sensitivity for Os isotope ratios by magnetic sector ICP-MS with a capacitive decoupling Pt guard electrode.
A magnetic sector ICP-MS with enhanced sensitivity was used to measure Os isotope ratios in solutions of low Os concentration (approximately 1 ng g(-1) or less). Ratios with 192Os as the basis were determined, while the geologically useful 187Os/188Os ratio was also measured. Sample introduction was via the traditional nebuliser-spray chamber method. A capacitive decoupling Pt shield torch was developed "in-house" and was found to increase Os signals by approximately 5 x under "moderate" plasma conditions (1050 W) over that found during normal operation (1250 W). Sensitivity using the guard electrode for 192Os was approximately 250-350,000 counts s(-1) per ng g(-1) Os. For a I ng g(-1) Os solution with no guard electrode, precisions of the order of 0.2-0.3% (189Os/192Os and 190Os/192Os) to approximately 1% or greater (186Os/192Os, 187Os/192Os and 187Os/188Os) were found (values as 1 sigma for n = 10). With the guard electrode in use, ratio precisions were found to improve to 0.2 to 0.8%. The total amount of Os used in the acquisition of this data was approximately 2.5 ng per measurement per replicate. At the higher concentration of 10 ng g(-1), precisions of the order of 0.15-0.3% were measured (for all ratios), irrespective of whether the shield torch was used. Ratio accuracy was confirmed by comparison with independently obtained NTIMS data. For both Os concentrations considered, the improvement in precision offered by the guard electrode (if any) was small in comparison to calculated theoretical values based on Poisson counting statistics, suggesting noise contributions from other sources (such as the sample introduction system, plasma flicker etc). At lower Os concentrations (to 100 pg g(-1)) no appreciable loss of ratio accuracy was observed, although as expected based on counting statistics, poorer precisions of the order of 0.45-3% (1 sigma, n = 5) were noted. Re was found to have a detrimental effect on the precision of Os ratios involving 187Os, indicating that separation of Re and Os samples is a necessary pre-requisite for highly accurate and precise Os isotope ratio measurements.


Association of age and social class with suicide among men in Great Britain.
The aim was to investigate suicide and "undetermined" deaths by age, economic activity status, and social class in Great Britain among males of working age. The study was a cross sectional analysis of Registrar General's data for England and Wales around 1981, repeated for around 1971, and for Scotland around 1971 and 1981. For England and Wales around 1971, suicide and undetermined death rates showed a progressive increase with age and a markedly higher rate in the lower social classes. A significant interaction effect was identified in the central age groups of the lower occupational categories. This interaction was confirmed in the remaining three data sets, notwithstanding some differences in the profile of age specific mortality. Other findings included a higher standardised mortality ratio for the economically inactive, who also showed an earlier peak in age specific mortality, and a relative concentration of undetermined as compared to suicide deaths in the lower social classes, but not all these further results were fully replicated. There is a concentration of suicide and undetermined deaths in the middle age groups of the lower social classes. Plausible explanations include both the social drift and the social genesis hypotheses, the latter including the effects of long term unemployment.