Behavior and therapeutic efficacy of beta-glucuronidase-positive mononuclear phagocytes in a murine model of mucopolysaccharidosis type VII.
Bone marrow transplantation (BMT) is relatively effective for the treatment of lysosomal storage diseases. To better understand the contribution of specific hematopoietic lineages to the efficacy of BMT, we transplanted beta-glucuronidase-positive mononuclear phagocytes derived from either the peritoneum or from bone marrow in vitro into syngeneic recipients with mucopolysaccharidosis type VII (MPS VII). Cell surface marking studies indicate that the bone marrow-derived cells are less mature than the peritoneal macrophages. However, both cell types retain the ability to home to tissues rich in cells of the reticuloendothelial system after intravenous injection into MPS VII mice. The half-life of both types of donor macrophages is approximately 7 days, and some cells persist for at least 30 days. In several tissues, therapeutic levels of beta-glucuronidase are present, and histopathologic analysis demonstrates that lysosomal storage is dramatically reduced in the liver and spleen. Macrophages intravenously injected into newborn MPS VII mice localize to the same tissues as adult mice but are also observed in the meninges and parenchyma of the brain. These data suggest that macrophages play a significant role in the therapeutic efficacy of BMT for lysosomal storage diseases and may have implications for treatments such as gene therapy.


Pharmacodynamics of PEG-IFN-[alpha]-2a and HCV response as a function of IL28B polymorphism in HIV/HCV-coinfected patients.
We examined the association between IL28B single-nucleotide polymorphism rs12979860, hepatitis C virus (HCV) kinetic, and pegylated interferon alpha-2a pharmacodynamic parameters in HIV/HCV-coinfected patients from South America. Twenty-six subjects received pegylated interferon alpha-2a + ribavirin. Serum HCV-RNA and interferon concentrations were measured frequently during the first 12 weeks of therapy and analyzed using mathematical models. African Americans and whites had a similar distribution of IL28B genotypes (P = 0.5). The IL28B CC genotype was overrepresented (P = 0.015) in patients infected with HCV genotype-3 compared with genotype-1. In both genotype-1 and genotype-3, the first-phase viral decline and the average pegylated interferon-alpha-2a effectiveness during the first week of therapy were larger (trend P <= 0.12) in genotype-CC compared with genotypes-TC/TT. In genotype-1 patients, the second slower phase of viral decline (days 2-29) and infected cells loss rate, [delta], were larger (P = 0.02 and 0.11, respectively) in genotype-CC than in genotypes-TC/TT. These associations were not observed in genotype-3 patients.


Cancer-associated adipocytes release FUCA2 to promote aggressiveness in TNBC.
Cancer-associated adipocytes (CAAs) have been suggested to promote tumor progression. Yet, the role of CAAs in triple-negative breast cancer (TNBC) is poorly investigated. We compared the expression of secretory protein-encoding genes in CAAs and control adipocytes. The effect of key secretory protein(s) on TNBC cell behaviors was explored. CAAs expressed and secreted FUCA2 at greater levels than control adipocytes. When FUCA2 activity was blocked with a neutralizing antibody, TNBC cell proliferation and migration induced by CAA-conditioned medium was impaired. In contrast, supplement of exogenous FUCA2 protein reinforced the proliferation, colony formation, and migration of TNBC cells. In vivo studies confirmed that FUCA2 exposure enhanced tumorigenesis and metastasis of TNBC cells. Mechanistic investigation revealed that FUCA2 induced TNBC aggressiveness through TM9SF3-dependent signaling. Depletion of TM9SF3 blocked CAA- and FUCA2-induced TNBC cell proliferation and migration. Compared to adjacent breast tissues, TNBC tissues had increased expression of TM9SF3. Moreover, high TM9SF3 expression was associated with advanced TNM stage, lymph node metastasis, and shorter overall survival of TNBC patients. Altogether, CAAs secrete FUCA2 to promote TNBC growth and metastasis through interaction with TM9SF3. Inhibition of TM9SF3 may represent a potential therapeutic strategy in the treatment of TNBC.


Molecular Docking analysis of the TNIK Receptor protein with a potential Inhibitor from the NPACT databas.
It is of interest to design and develop efficient inhibitors to the TNIK protein target in Wnt signaling pathways in the context of colorectal cancer (CRC) using molecular docking models. We show data to support that a compound named aglafoline (methyl (1R,2R,3S,3aR,8bS)-1,8b-dihydroxy-6,8-dimethoxy-3a-(4-methoxyphenyl)-3-phenyl-2,3-dihydro-1H cyclopenta [b] [1] from the NPACT (Naturally Occurring Plant-based Anti-cancer Compound-Activity-Target database) database have optimal binding features with the TNIK receptor for further consideration in this context.


Arsenic metabolism and cancer risk: A meta-analysis.
To describe the studies that have reported association measures between risk of cancer and the percentage distribution of urinary inorganic arsenic (iAs) metabolites by anatomical site, in non-ecological epidemiological studies. Studies were identified in the PubMed database in the period from 1990 to 2015. Inclusion criteria were: non-ecological epidemiological study, with histologically confirmed cancer cases, reporting the percentage distribution of inorganic arsenic (iAs), monomethylated (MMA) and dimethylated (DMA) metabolites, as well as association measures with confidence intervals (CI) between cancer and %iAs and/or %MMA and/or %DMA. A descriptive meta-analysis was performed by the method of the inverse of the variance for the fixed effects model and the DerSimonian and Laird's method for the random effects model. Heterogeneity was tested using the Q statistic and stratifying for epidemiological design and total As in urine. The possibility of publication bias was assessed through Begg's test. A total of 13 eligible studies were found, most of them were performed in Taiwan and focused on skin and bladder cancer. The positive association between %MMA and various types of cancer was consistent, in contrast to the negative relationship between %DMA and cancer that was inconsistent. The summary risk of bladder (OR=1.79; 95% CI: 1.42, 2.26, n=4 studies) and lung (OR=2.44; 95% CI: 1.57, 3.80, n=2 studies) cancer increased significantly with increasing %MMA, without statistical heterogeneity. In contrast, lung cancer risk was inversely related to %DMA (OR=0.58; 95% CI: 0.36, 0.93, n=2 studies), also without significant heterogeneity. These results were similar after stratifying by epidemiological design and total As in urine. No evidence of publication bias was found. These findings provide additional support that methylation needs to be taken into account when assessing the potential iAs carcinogenicity risk.


Safety and tolerability of HIV-1 multiantigen pDNA vaccine given with IL-12 plasmid DNA via electroporation, boosted with a recombinant vesicular stomatitis virus HIV Gag vaccine in healthy volunteers in a randomized, controlled clinical trial.
The addition of plasmid cytokine adjuvants, electroporation, and live attenuated viral vectors may further optimize immune responses to DNA vaccines in heterologous prime-boost combinations. The objective of this study was to test the safety and tolerability of a novel prime-boost vaccine regimen incorporating these strategies with different doses of IL-12 plasmid DNA adjuvant. In a phase 1 study, 88 participants received an HIV-1 multiantigen (gag/pol, env, nef/tat/vif) DNA vaccine (HIV-MAG, 3000 μg) co-administered with IL-12 plasmid DNA adjuvant at 0, 250, 1000, or 1500 μg (N = 22/group) given intramuscularly with electroporation (Ichor TriGrid™ Delivery System device) at 0, 1 and 3 months; followed by attenuated recombinant vesicular stomatitis virus, serotype Indiana, expressing HIV-1 Gag (VSV-Gag), 3.4 ⊆ 107 plaque-forming units (PFU), at 6 months; 12 others received placebo. Injections were in both deltoids at each timepoint. Participants were monitored for safety and tolerability for 15 months. The dose of IL-12 pDNA did not increase pain scores, reactogenicity, or adverse events with the co-administered DNA vaccine, or following the VSV-Gag boost. Injection site pain and reactogenicity were common with intramuscular injections with electroporation, but acceptable to most participants. VSV-Gag vaccine often caused systemic reactogenicity symptoms, including a viral syndrome (in 41%) of fever, chills, malaise/fatigue, myalgia, and headache; and decreased lymphocyte counts 1 day after vaccination. HIV-MAG DNA vaccine given by intramuscular injection with electroporation was safe at all doses of IL-12 pDNA. The VSV-Gag vaccine at this dose was associated with fever and viral symptoms in some participants, but the vaccine regimens were safe and generally well-tolerated. Clinical Trials.gov NCT01578889.


Epidermal and fibroblast growth factors enhance fibrovascular integration of porous polyethylene implants.
Porous implants used in functional and aesthetic reconstruction of the orbit, face, and cranium are less likely to develop complications after they become biointegrated. We investigated whether the administration of exogenous growth factors could increase the rate of implant integration. High-density porous polyethylene cubes were placed in dorsal paraspinal muscles of rabbits, and daily transcutaneous injections of saline, epidermal growth factor, or basic fibroblast growth factor were administered directly over the cubes for 10 days. At serial time points up to 10 weeks, cubes were explanted and the fibroblasts present at the center of the cubes were counted. Injections of epidermal growth factor and basic fibroblast growth factor increased the rate at which fibroblasts accumulated in porous polyethylene implants and decreased the time required to achieve a maximal rate of cellular accumulation within the cubes. At 4 weeks, when all cell populations had attained a linear rate of accumulation, cubes previously injected with saline, epidermal growth factor, or basic fibroblast growth factor contained an average of 10, 40, and 80 cells per 0.0156 mm2, at their centers, respectively. Enhancement of the rate of biointegration of porous polyethylene cubes in rabbits is achievable by repeated, transcutaneous administration of exogenous growth factors.


Inbreeding depresses sperm competitiveness, but not fertilization or mating success in male Tribolium castaneum.
As populations decline to levels where reproduction among close genetic relatives becomes more probable, subsequent increases in homozygous recessive deleterious expression and/or loss of heterozygote advantage can lead to inbreeding depression. Here, we measure how inbreeding across replicate lines of the flour beetle Tribolium castaneum impacts on male reproductive fitness in the absence or presence of male-male competition. Effects on male evolution from mating pattern were removed by enforcing monogamous mating throughout. After inbreeding across eight generations, we found that male fertility in the absence of competition was unaffected. However, we found significant inbreeding depression of sperm competitiveness: non-inbred males won 57 per cent of fertilizations in competition, while inbred equivalents only sired 42 per cent. We also found that the P(2) 'offence' role in sperm competition was significantly more depressed under inbreeding than sperm 'defence' (P(1)). Mating behaviour did not explain these differences, and there was no difference in the viability of offspring sired by inbred or non-inbred males. Sperm length variation was significantly greater in the ejaculates of inbred males. Our results show that male ability to achieve normal fertilization success was not depressed under strong inbreeding, but that inbreeding depression in these traits occurred when conditions of sperm competition were generated.


Photoconductivity Response at Cyclotron-Resonance Harmonics in a Nondegenerate Two-Dimensional Electron Gas on Liquid Helium.
We report the first observation of an oscillatory photoconductivity response at the cyclotron-resonance harmonics in a nondegenerate 2D electron system formed on the free surface of liquid helium. The dc conductivity oscillations are detected for electrons occupying the ground surface subband. Their period is governed by the ratio of the microwave frequency to the cyclotron frequency. Theoretical analysis of the photoresponse in a strongly interacting electron system indicates that the observation can be explained by an oscillatory correction to the electron distribution function that appears for a large inelastic relaxation time because of photon-assisted scattering.


Investigation on the Status and Determinants of Caregiver Burden on Caring for Patients with Chronic Wound.
Objective: To investigate the caring burden and its determinants of primary informal caregiver of patients with chronic wound. Approach: A prospective cross-sectional study of 132 pairs of chronic wound care recipients and their informal caregivers was included. The characteristics of patients and their informal caregivers as well as caregiver burden assessment by the caregiver burden inventory (CBI) were measured. Single factor analysis and multiple regression analysis were carried out to explore the independent determinants of caregiver burden on caring for patients with chronic wound. Results: Most of the caregivers were female with mean age of 54.57 ± 13.35 years, and 58.3% of the caregivers were adult children. The mean CBI score was 34.21 ± 9.69 at a medium level. The following variables increased the CBI scores of caregivers: long caring time per day for patients, powerlessness status of patients, insufficient self-efficacy, and social support of caregivers, the model was able to explain 67.5% of variance in caregiver burden (F = 47.167, p = 0.000, R 2 = 0.675, adjusted R 2 = 0.660). Innovation: Caring burden of patients with chronic wound as a key consideration of patient-centered wound care has received relatively little attention. In this study we report the status of caring burden and reveal its determinants of primary informal caregiver of patients with chronic wound. Conclusion: Wound professionals are suggested to pay attention to the caregiver burden of patients with chronic wound and develop family-centered intervention support service system according to the determinants of caregiver burden to alleviate the caregiver burden.


Predictors of Human Papillomavirus Vaccination in Female University Students.
The purpose of this study was to identify predictors of HPV (Human papillomavirus) vaccination among female university students. The participants in this study were 1,486 female university students. Data were collected using self-administered questionnaire from september 1, 2010 to June 30, 2011. Data were analysed using descriptive statistics, t-test, chi2-test and logistic regression. The rate of HPV vaccination was 7.2%. Factors that influence HPV vaccination included residential region, smoking experience, family history of cervical cancer, experience having heard about the HPV vaccination, awareness that HPV is a sexual disease, HPV knowledge, perceived seriousness, perceived benefit and perceived barrier in health beliefs. The results suggest a need to increase HPV vaccination campaigns to increase awareness of HPV vaccination. University and health care facilities should provide health education emphasizing perceived seriousness and perceived benefits. Also, the national health insurance corporation should consider addressing cost issues as a barrier to HPV vaccination.


Multimodality treatment of cerebral AVMs in children: a single-centre 20 years experience.
The purpose of this study was to review our experience with a multimodality treatment approach in the management of cerebral arteriovenous malformation (AVM) in children. We retrospectively analysed a consecutive series of 56 children who harboured a cerebral AVM and were treated at our institution between 1988 and 2008. During the whole treatment period, a combined treatment strategy, including microsurgery, endovascular treatment and gamma knife radiosurgery, was used. Of the 56 patients (median age, 14.0; range, 3 months-18 years) reported, 36 (64.3%) were admitted after AVM rupture; of these, only one AVM (1.8%) was considered untreatable. In 30.9% (17/55) of the treated patients, a single treatment measure was sufficient to attain angiographic cure of the AVM. Among these, six patients (10.9%) had microsurgical AVM resection, four patients (7.3%) underwent endovascular treatment, and another seven patients (12.7%) underwent radiosurgical management of the AVM. The majority of the population (38/55; 69.1%) underwent combined treatment: 21 patients (38.2%) received embolisation followed by radiosurgery of the remnant nidus, ten patients (18.2%) underwent embolisation with subsequent surgical resection of the residual AVM, three patients (5.5%) had radiosurgery after incomplete surgical AVM nidus resection and another four patients (7.3%) required a combination of all three treatment modalities to achieve permanent angiographic cure of the AVM. We observed good clinical outcomes (Glasgow Outcome Scale 5 and 4) in 94.6% of the children. Complete angiographic obliteration was achieved in 93.3% of the patients treated. A multimodality treatment approach in children harbouring cerebral AVMs leads to excellent angiographic and clinical outcomes.


Antioxidant Capacities of Plant-Derived Foods Commonly Consumed in Japan.
To provide reliable data for high quality epidemiological studies examining the relationship between health and antioxidant intake from daily foods, 107 plant-derived food items (12 rice, bread and noodles, 5 potatoes and starches, 9 pulses, 6 nuts/seeds, 29 vegetables, 22 fruits, 5 mushrooms, 7 algae, and 12 beverages) were selected as commonly consumed foods in Japan based on dietary records, and their antioxidant capacities were evaluated by validated hydrophilic- and lipophilic-oxygen radical absorbance capacity (H-ORAC and L-ORAC) methods. The food items covered more than 60% of total food intake for each category on a weight basis. The H-ORAC and L-ORAC values were widely distributed at 0-210 and 0-30 μmol-Trolox equivalent/g, respectively. The foods possessing potent antioxidant capacities were found in vegetables and fruits as well as other plant-derived foods. In most foods measured, the H-ORAC values were much larger than the L-ORAC values, except for certain kinds of pulses, nuts/seeds, mushrooms, and algae. The ORAC data shown here is sufficient to accurately estimate the antioxidant intake from plant-derived foods in Japan, and should be useful in future epidemiological studies aiming to clarify the biological significance of ORAC values.


Multicenter evaluation of BD Veritor System and RSV K-SeT for rapid detection of respiratory syncytial virus in a diagnostic laboratory setting.
The recently introduced BD Veritor System RSV laboratory kit (Becton Dickinson, Sparks, MD, USA) with automatic reading was evaluated and compared with the RSV K-SeT (Coris BioConcept, Gembloux, Belgium) for the detection of respiratory syncytial virus (RSV) using 248 nasopharyngeal aspirates of children younger than 6 years old with respiratory tract infection. Compared to reverse transcriptase polymerase chain reaction as gold standard, both tests had an identical sensitivity of 78.1% and a specificity of 96.8% and 95.8% for the BD Veritor System and RSV K-SeT, respectively. Both antigen tests can be used to reliably confirm RSV in young children. However, a negative result does not definitively exclude the presence of RSV.


Using Optimal Transport to Improve Spherical Harmonic Quantification of Complex Biological Shapes.
The knowledge of the anatomical shape of both gross and microscopic structures is the key to understanding the effects of disease processes on cellular structure. Geometric morphometric methods, such as Procrustes superimposition, and Spherical Harmonics (SPHARM), have been used to capture the biological shape variation and group differences in morphology. Previous SPHARM-MAT techniques use the CALD algorithm to parameterize the mesh surface. It starts from initial mapping and performs local and global smoothing methods alternately to control the area and length distortions simultaneously. However, this parameterization may not be sufficient in complex morphological cases. To bridge this gap, we propose SPHARM-OT, an enhanced SPHARM surface modeling method using optimal transport (OT) for spherical parameterization. First, the genus 0 3D objects are conformally mapped onto a sphere. Then the optimal transport theory via spherical power diagram is introduced to minimize the area distortion. This new algorithm can effectively reduce the area distortion and lead to a better reconstruction result. We demonstrate the effectiveness of the method by applying it to the human sphenoidal paranasal sinuses.


SARS-CoV-2-Specific Memory T Lymphocytes From COVID-19 Convalescent Donors: Identification, Biobanking, and Large-Scale Production for Adoptive Cell Therapy.
Syndrome coronavirus 2 (SARS-CoV-2) pandemic is causing a second outbreak significantly delaying the hope for the virus' complete eradication. In the absence of effective vaccines, we need effective treatments with low adverse effects that can treat hospitalized patients with COVID-19 disease. In this study, we determined the existence of SARS-CoV-2-specific T cells within CD45RA- memory T cells in the blood of convalescent donors. Memory T cells can respond quickly to infection and provide long-term immune protection to reduce the severity of COVID-19 symptoms. Also, CD45RA- memory T cells confer protection from other pathogens encountered by the donors throughout their life. It is of vital importance to resolve other secondary infections that usually develop in patients hospitalized with COVID-19. We found SARS-CoV-2-specific memory T cells in all of the CD45RA- subsets (CD3+, CD4+, and CD8+) and in the central memory and effector memory subpopulations. The procedure for obtaining these cells is feasible, easy to implement for small-scale manufacture, quick and cost-effective, involves minimal manipulation, and has no GMP requirements. This biobank of specific SARS-CoV-2 memory T cells would be immediately available "off-the-shelf" to treat moderate/severe cases of COVID-19, thereby increasing the therapeutic options available for these patients.


Uterine prolapse associated with bladder exstrophy: surgical management and subsequent pregnancy.
Congenital bladder exstrophy affects 1 in 125,000 to 250,000 females. Consisting of absence of the anterior abdominal wall with exposure of the ureteral orifices, failure of pubic symphysis fusion, and deficient anterior pelvic diaphragm musculature, bladder exstrophy is frequently associated with genital prolapse. Pregnancy may be complicated by recurrent urinary tract infections, preterm labor, mild procidentia, and malpresentation. Due to the rarity of the condition, there is a corresponding scarcity of obstetric literature regarding management during pregnancy. We report the case of a young woman with surgically repaired bladder exstrophy who developed genital prolapse. The uterus was suspended using a sacral colpopexy utilizing a Gore-Tex graft. Subsequently, the patient became pregnant and delivered a healthy male infant at 35 weeks' gestation via cesarean section (without recurrence of the genital prolapse postpartum). Sacral colpopexy to correct genital prolapse associated with bladder exstrophy may preserve fertility in young patients.


Presence of the variant mannose-binding lectin alleles associated with slower progression to AIDS. Amsterdam Cohort Study.
To examine the association between mannose-binding lectin (MBL) polymorphism and progression to AIDS and death in HIV-1 infection. In 131 HIV-1-infected homosexual seroconverters, survival analyses were performed to determine both the association between MBL genotype and time from HIV-1 seroconversion to AIDS and death, and time from AIDS to death. Of the 131 seroconverters, of whom 61 developed AIDS, 76 were typed as homozygous wild-type and 55 as carriers of variant alleles (52 heterozygous and three homozygous variant alleles). A Survival analyses suggested that HIV-1-infected men with the variant alleles progressed somewhat slower to AIDS [relative hazard (RH), 0.62; 95% confidence interval (CI), 0.36-1.10] and death (RH, 0.73; 95% CI, 0.42-1.25). Interestingly, CD4+ T-cell count determined at the moment of AIDS was found to be significantly lower among persons with the mutation (97 x 10(6)/l versus 204 x 10(6)/l; P=0.03). Furthermore, when AIDS-free times before the diagnosis of an opportunistic infection were compared with those preceding a diagnosis of Kaposi's sarcoma, Kaposi's sarcoma diagnosis was more postponed than that of an opportunistic infection (RH, 0.21; 95% CI, 0.05-0.95; versus RH, 0.67; 95% CI, 0.35-1.27). Indications for a weak pre-AIDS protective effect of variant MBL alleles were demonstrated.


Advances in multimodality imaging through a hybrid PET/MRI system.
The development of integrated imaging systems for magnetic resonance imaging (MRI) and positron emission tomography (PET) is currently being explored in a number of laboratories and industrial settings. PET/MRI scanners for both preclinical and human research applications are being developed. PET/MRI overcomes many limitations of PET/computed tomography (CT), such as limited tissue contrast and high radiation doses delivered to the patient or the animal being studied. In addition, recent PET/MRI designs allow for simultaneous rather than sequential acquisition of PET and MRI data, which could not have been achieved through a combination of PET and CT scanners. In a combined PET/CT scanner, while both scanners share a common patient bed, they are hard-wired back-to-back and therefore do not allow simultaneous data acquisition. While PET/MRI offers the possibility of novel imaging strategies, it also creates considerable challenges for acquiring artifact-free images from both modalities. In this review, we discuss motivations, challenges, and potential research applications of developing PET/MRI technology. A brief overview of both MRI and PET is presented and preclinical and clinical applications of PET/MRI are identified. Finally, issues and concerns about image quality, clinical practice, and economic feasibility are discussed.


Mutations of two acidic residues at the cytoplasmic end of segment IIIS6 of an insect sodium channel have distinct effects on pyrethroid resistance.
Mutations in sodium channels are known to confer knockdown resistance to pyrethroid insecticides, such as permethrin and cypermethrin, in various agricultural pests and disease vectors. Double mutations, D3i28V and E3i32G, were detected in cypermethrin-resistant Helicoverpa armigera and Heliothis virescens populations. However, the role of the two mutations in pyrethroid resistance remains unclear. In this study, we introduced the mutations into the cockroach sodium channel, BgNav1-1a, and examined their effects on channel gating and pyrethroid sensitivity in Xenopus oocytes. D3i28V alone and the double mutation, D3i28V/E3i32G, shifted the voltage dependence of activation in the depolarizing direction by 15 mV and 20 mV, respectively, whereas E3i32G had no significant effect. D3i28V reduced the amplitude of tail currents induced by permethrin and NRDC 157 (Type I pyrethroids) and deltamethrin and cypermethrin (Type II pyrethroids), whereas E3i32G alone had no effect. Intriguingly, the amplitude of Type II pyrethroid-induced tail current from D3i28V/E3i32G channels was similar to that of BgNav1-1a channels, but the decay of the tail currents was accelerated. Such effects were not observed for Type I pyrethroid-induced tail currents. Computational analysis based on the model of dual pyrethroid receptors on insect sodium channels predicted D3i28V and E3i32G exert their effects on channel gating and pyrethroid action via allosteric mechanisms. Our results not only illustrate the distinct effect of the D3i28V/E3i32G double mutations on Type I vs. Type II pyrethroids, but also reinforce the concept that accelerated decay of tail currents can be an effective mechanism of pyrethroid resistance to Type II pyrethroids.


High resolution crystal structure of BRD4-BD1 in complex with a novel inhibitor precursor.
The inhibition of BRD4 bromodomain is an effective therapeutic strategy for a variety of diseases in which BRD4 are implicated. Herein, we identified a small-molecule BRD4 inhibitor hit named compound 3 using high-throughput screening. The 1.6 Å resolution co-crystal structure confirmed that the compound occupies the KAc recognition pockets of BRD4 by forming key hydrogen bonds with Asn140 and engaging in hydrophobic interactions, thus impedes the binding of acetylated lysine to BRD4. These findings suggest compound 3 can be a lead compound to develop a structurally novel BRD4 inhibitors.


Evaluating the role of strain ratio elastography in determining malignancy potential and calculating objective BIRADS US scores using ultrasonography and elastography features.
The aims of this study were, firstly, to evaluate solid breast masses based on their malignancy potential and to determine whether the strain elastography ratio (SER) can contribute to classical grey-scale ultrasonography findings, and secondly, to define objective BIRADS US scores using ultrasound (US) and SER findings. A total of 280 patients and 297 solid breast masses were evaluated using sonographic and elastographic data. The SER was measured for each lesion. The positive predictive values (PPV) for each criterion was calculated to be between 35% and 83.3%. The lowest PPV was obtained from hypoechogenicity (35%) and the highest PPV was obtained for anti-parallel features (83.3%). The difference between the mean SER of benign and malignant lesions was statistically significant. After ROC analysis, the SER cut-off value was calculated to be 3.1 for determining if the mass was benign or malignant. Mass scores were calculated for each solid breast mass based on positive predictive values, and BIRADS US score was defined as the sum of mass scores. SER findings can be used as malignancy criteria in evaluating solid breast masses. BIRADS US score can be objectively determined based on US and elastography features instead of doing subjective scoring. As an additional result, all solid breast masses have the possibility to be malignant, even though US and elastography findings indicate the opposite.


Novel COL4A1 mutations associated with HANAC syndrome: a role for the triple helical CB3[IV] domain.
The COL4A1 gene encodes the α1-chain of type IV collagen, which is ubiquitously expressed in basement membranes. Mutations in COL4A1 have been reported in autosomal-dominant porencephaly and in patients with symptomatic small vessel brain disease, inconstantly associated with eye defects. We have previously reported three COL4A1 mutations associated with a systemic phenotype that we called HANAC (Hereditary Angiopathy, Nephropathy, Aneurysms, and Cramps). We carried out a clinical and genetic study of three families presenting with characteristic features of HANAC syndrome. Common systemic signs included arterial retinal tortuosity and muscle cramps, with a variable combination of small vessel brain disease, Raynaud phenomena, and kidney defects. Three novel COL4A1 missense substitutions are described, which affect highly conserved glycine residues within the collagenous domain of the protein. All six known mutations associated with the HANAC phenotype are localized within the CB3[IV] fragment of COL4A1, which encompasses major integrin-binding sites. Our results confirm that HANAC syndrome is a distinct clinical entity within the COL4A1-related disorders, which is characterized by systemic involvement and usually asymptomatic brain disease. The restricted distribution of COL4A1 mutations within the CB3[IV] region is a characteristic of the reports of patients with HANAC, which suggests that abnormal cell-type IV collagen interactions may underlie the systemic defects observed in this syndrome.


Functional mapping of the orientation for TCR recognition of an H2-Kb-restricted ovalbumin peptide suggests that the beta-chain subunit can dominate the determination of peptide side chain specificity.
T cells recognize a complex of antigenic peptide bound to the class I or class II products of the MHC. Crystallographic analysis of the interaction between MHC class I-bound peptide fragments and specific TCR have recently been described and highlight the importance of the CDR3 in determining peptide specificity. The results presented here show functional data for TCR recognition of the H2-Kb class-I restricted determinant derived from OVA (SIINFEKL) that are consistent with the TCR orientation defined by these crystal structures. In addition, we also found that the beta-chain CDR3 dominates side chain specificity for the most exposed regions within this peptide. The data also suggest that this orientation and pattern of beta-chain dominance may extend to the recognition of a second H2-Kb-restricted peptide from the herpes simplex virus type 1 glycoprotein B (SSIEFARL), which shares a common alpha-chain contact with the OVA peptide. These results are discussed in terms of a common orientation for TCR-ligand interaction and the greater potential for TCR beta-chain CDR3 diversity in determining peptide side chain specificity.


Patterns of chemotherapy use for women with ovarian cancer: a population-based study.
To evaluate adherence to published recommendations for chemotherapy for ovarian cancer patients in the general community and to identify factors associated with its use. The study population consisted of 2,150 women residing in Northern California with a first diagnosis of primary epithelial ovarian cancer between January 1994 and December 1996. Patients were identified through the California Cancer Registry and their physicians were surveyed to supplement registry treatment information. Almost 89% of women younger than 75 years with International Federation of Gynecology and Obstetrics stage III or IV tumors received chemotherapy, with levels of treatment highest for women diagnosed at stage III. Patients 75 years of age and older were significantly less likely than younger women to receive chemotherapy (58.2% v 86.1%; P =.001) regardless of stage at diagnosis. Approximately 20% of patients younger than 55 years with early-stage (stage IC and II) cancer received no chemotherapy. Treatment in an American College of Surgeons hospital and treatment by a gynecologic oncologist increased the likelihood of receiving chemotherapy. Hospitalization for comorbid illness, race/ethnicity, census-based measures of socioeconomic status, and size or teaching status of hospital were all unrelated to probability of treatment after adjustment for other factors. Reasons reported most frequently by physicians for no treatment were lack of clinical indication and patient refusal. The results of this study suggest that, despite scientific evidence and published guidelines that advocate chemotherapy for most women with ovarian cancer, some groups of women did not receive optimum treatment.


Ube3a imprinting impairs circadian robustness in Angelman syndrome models.
The paternal allele of Ube3a is silenced by imprinting in neurons, and Angelman syndrome (AS) is a disorder arising from a deletion or mutation of the maternal Ube3a allele, which thereby eliminates Ube3a neuronal expression. Sleep disorders such as short sleep duration and increased sleep onset latency are very common in AS. We found a unique link between neuronal imprinting of Ube3a and circadian rhythms in two mouse models of AS, including enfeebled circadian activity behavior and slowed molecular rhythms in ex vivo brain tissues. As a consequence of compromised circadian behavior, metabolic homeostasis is also disrupted in AS mice. Unsilencing the paternal Ube3a allele restores functional circadian periodicity in neurons deficient in maternal Ube3a but does not affect periodicity in peripheral tissues that are not imprinted for uniparental Ube3a expression. The ubiquitin ligase encoded by Ube3a interacts with the central clock components BMAL1 and BMAL2. Moreover, inactivation of Ube3a expression elevates BMAL1 levels in brain regions that control circadian behavior of AS-model mice, indicating an important role for Ube3a in modulating BMAL1 turnover. Ube3a expression constitutes a direct mechanistic connection between symptoms of a human neurological disorder and the central circadian clock mechanism. The lengthened circadian period leads to delayed phase, which could explain the short sleep duration and increased sleep onset latency of AS subjects. Moreover, we report the pharmacological rescue of an AS phenotype, in this case, altered circadian period. These findings reveal potential treatments for sleep disorders in AS patients.


Circular-dichroic properties and secondary structure of Pseudomonas aeruginosa soluble cytochrome c oxidase.
The c.d. spectra of Pseudomonas aeruginosa cytochrome c oxidase in the oxidized state and the reduced state are reported in the visible- and u.v. absorption regions. In the visible region the comparison between the spectra of reduced cytochrome c oxidase and ferrocytochrome c-551 allows the identification of the c.d. bands mainly due to the d1 haem chromophore in cytochrome c oxidase. In the near-u.v. region the assignment of some of the observed peaks to the haem groups and to the aromatic amino acid residues is proposed. A careful analysis of the data in the far-u.v. region leads to the determination of the relative amounts of alpha-helix and beta-sheet in the enzyme, giving for the first time a picture of its secondary structure. A significant difference in this respect between the reduced and the oxidized species is observed and discussed in the light of similar conclusions reported by other workers.


The effect of direct referral for fast CT scan in early lung cancer detection in general practice. A clinical, cluster-randomised trial.
This PhD thesis is based on the project "The effect of direct referral for fast CT scan in early lung cancer detection in general practice. A clinical, cluster-randomised trial", performed in Denmark in 2010-2013. The thesis includes four papers and focuses on early lung cancer diagnostics in general practice. A total of 4200 new cases of lung cancer are diagnosed in Denmark annually. The stage of the disease is an important prognostic factor; thus, the opportunity for curative treatment declines with more advanced tumour stage. Lung cancer patients in Denmark (like in the UK) have a poorer prognosis than lung cancer patients in other European countries. One explanation could be delayed diagnosis. A fast-track pathway was therefore introduced in an attempt to expedite the diagnosis of cancer. However, it seems that not all patients can be diagnosed through this pathway. In order to ensure fast and early lung cancer diagnosis, it is crucial to examine the initial diagnostic process in general and the role general practice plays in lung cancer diagnostics in particular. The specific areas of investigation include the pathways to diagnosis, the characteristics of patients who are at special risk of delayed diagnosis and the level of prediagnostic activity in general practice. A chest radiograph is often the first choice in the investigation of lung cancer. Unfortunately, radiographs are less suitable for central and small tumours. Low-dose computer tomography (LDCT), however, has a high sensitivity for lung cancer which implies that it can be used to detect patients with localised, potentially curable disease. The aim of this thesis was to increase our knowledge of the initial stages of lung cancer diagnostics in general practice. The thesis also examined the effect of a direct referral from general practice to an additional diagnostic test, the LDCT. The aims of this thesis were: 1) To describe Danish patients' pathways to the diagnosis of lung cancer in general and the prediagnostic activity leading up to diagnosis in particular. An additional aim was to explore the diagnostic intervals for specific patient groups (Paper I). 2) In a randomised, controlled trial including all patients referred for the existing fast-track scheme to either direct chest and upper abdomen CE-MDCT or to evaluation by the chest physician, (i) to test: Fast-track performance measured by the number of CE-MDCT scans and chest physician specialist time per diagnosis (Paper II) 3) In a two-arm, clinical, controlled, cluster-randomised trial where direct referral to CT together with a lung cancer update is compared with usual practice, (i) to test how CT is used in this group of patients and the outcome of CT (Paper III); and (ii) to test the effect of either modality on the time to lung cancer diagnosis, the TNM stage and the use of the fast-track pathway for lung cancer (Paper IV). Study I was a national registry-based cohort study of 971 consecutive, incident lung cancer patients in 2010 Data were derived from national registries and questionnaires filled in by general practitioners (GPs). Study II was a randomised, controlled trial enrolling 493 patients referred from general practice to a fast-track evaluation. Half of the patients were randomly assigned to the intervention and went straight to a chest CT before a chest physician evaluation. Studies III and IV were a cluster-randomised, controlled trial (IV) and a cohort study nested in the trial (III). A total of 199 general practices with 266 GPs were randomised into two groups. Intervention GPs were offered direct access to a low-dose chest CT combined with a meeting on early lung cancer detection. Study III concerned the intervention arm solely and reported uses and outcomes of the scans. Study IV evaluated the effect of direct low-dose CT on the time to diagnosis and stage at diagnoses for patients from intervention and control GPs. In Study I, we found that GPs were involved in 2/3 of all lung cancer diagnostic pathways. One quarter of the patients followed the obvious pathway from general practice to fast-track detection. At least one radiograph was performed in 85.6% of patients, whereas 1/3 of all patients had two or more radiograph performed during the 90 days preceding diagnosis. Patients with co-morbidity or unspecific symptoms more often had two or more X-rays performed than patients without these characteristics. In Study II, there was no difference between the groups in the number of CTs performed. In the intervention group, chest physicians spent mean 13.3 minutes less per referred patient than in the control group. In Study III, we found that 648 patients were referred to low-dose CT during a 19-month period. Half of the referred patients needed further work-up, and 15 (2.3%) of the patients had lung cancer, 60% in a localised stage. For all patients, 6.8% were diagnosed with a severe lung disease. In all, 2/3 of the GPs used the CT opportunity; and the referral rate was 61% higher for GPs participating in the lung cancer meeting than for GPs who did not participate in such meetings. In Study IV, we found that direct, low-dose CT from primary care did not significantly influence stage at diagnosis and had only a limited impact on time to diagnosis. This thesis contributes to the knowledge of the early diagnosis of lung cancer in Denmark. General practice was found to play an important role, but only a small part of Danish lung cancer patients were diagnosed from general practice through the fast-track pathway. This together with the fact that a high proportion of patients had two or more radiographs within the 90 days preceding the diagnosis indicate that other diagnostic strategies should be tested in an attempt to provide GPs with the best opportunity for early diagnosis. This thesis provides evidence that GPs are, indeed, able to refer patients straight-to-test in the fast-track pathway. This knowledge may be used when organising other fast tracks. Furthermore, GPs participating in education about early lung cancer diagnosis were willing to refer patients direct to low-dose CT (LDCT) from primary care. Half of the patients needed further diagnostic work-up, and 2.3% of all patients referred were diagnosed with lung cancer. In addition, many lung diseases were diagnosed by LDCT. No effect on time to diagnosis or stage at diagnosis was found when patients from intervention GPs were compared with patients from control GPs. The effect of combining direct access to LDCT with referral to the existing fast-track pathway should be analysed as it may ensure earlier and faster lung cancer detection in primary care. Direct access to LDCT scan may also be an alternative to lung cancer screening. Furthermore, if a LDCT screening program is going to be implemented, it should be considered to supplement the program with access to CT directly from primary care for the symptomatic, not-screened patients.


Diabetic retinopathy and pancreas transplantation: a 3-year follow-up.
The effect of simultaneous pancreas and kidney transplantation on diabetic retinopathy was studied in a prospective study with 30 patients (57 eyes) and 15 control subjects (26 eyes), patients who lost the pancreas, but preserved kidney function. There was no significant difference between the groups after a mean observation time of more than 35 months (a range of 12 to 96 months). Both populations had a stable retinopathy during follow-up. This seems to be a consequence of the far advanced retinopathy (mean duration of type 1 diabetes was 22 years) and the high percentage of coagulated eyes (81% and 85%, respectively), but is not related to the organ transplantation. A closer look at the few patients who did not receive laser coagulation (14 patient and 6 control eyes), produced a different result. Four control eyes experienced a significant deterioration of the retinopathy which had been stable before rejection. It is the most important and so far never mentioned aspect of this study, that periods of destabilisation are a definite threat for the retinopathy. Nevertheless, it seems questionable whether we will ever be able to make a definite statement on the pancreas-eye relation, as long as the transplantation must be restricted to carefully selected late-stage diabetic subjects.


Ultrafast optical pulse convertor caused by oscillations of the energy level structure in the conjugated polymer poly(p-phenylenevinylene).
For a conjugated polymer irradiated by two optical pulses, the whole process of excitation, involving lattice oscillations, oscillations of the energy level structure, and evolution of the electron cloud, is investigated. Localization of the electron cloud appears in the first 100 fs of irradiation, which in turn induces vibrations of lattice of the polymer chain as well as oscillations of the band gap. These oscillations filter the absorption of the external optical field inversely and convert the original optical field to an ultrafast light field whose intensity varies with a certain period. Based on the mechanism, oscillations of the energy level structure, induced by the external excitation, can be designed as an ultrafast response optical convertor that is able to change the external optical pulse into a new effective light field with a certain oscillation period. This helps provide new insight into designing nanostructures for polymeric optoelectronics.


Neglecting the call of the wild: Captive frogs like the sound of their own voice.
Acoustic communication is highly influential in the expression of social behavior by anuran amphibians, transmitting information about the individual's physical condition and motivation. We studied the phonotactic (approach movements) responses of wild and captive male golden mantella frogs to conspecific wild and captive playback calls to determine the impact of captivity on social behaviour mediated by vocalisations. Calls were recorded from one wild and two captive populations. Phonotaxis experiments were then conducted by attracting M. aurantiaca males across a PVC grid on the forest floor or enclosure floor to a speaker. For each playback, the following parameters were recorded to define the accuracy of phonotaxis: (1) number of jumps; (2) jump angles; (3) jump distances; (4) path straightness. During this experiment we observed that wild frogs had a similar behavioural (phonotaxis) response to calls independent of their source while frogs from Chester Zoo had a significantly stronger response to calls of other conspecifics held separately at Chester Zoo. The lack of appropriate phonotaxis response by captive bred frogs to the calls of wild conspecifics could have serious negative conservation implications, if the captive bred individuals were released back to the wild.


[Endovascular operations on leg arteries].
The paper presents the results color duplex scanning (CDS) diagnosis during 42 endovascular operations (40 balloon angioplasties and 2 stentings) on leg arteries with stenoocclusive changes. The operations were successful in 95.5% of cases, as evidenced by angiography and in 83.3% as shown by CDS; there were 7 (16.7%) cases of hemodynamic events, such as angioplastic site thrombosis, intimal dissection, and residual stenosis. Late follow-ups revealed 6 (16%) cases of hemodynamic events, such as angioplastic area thrombosis and reocclusion. CDS permitted evaluation of the patency of leg arterial endovascular intervention areas, which was 44.96+/-19.77% at a follow-up of up to 60 months (mean follow-up period 18.76+/-2.58). Two stents were patent within 24 months.


Dynamics of the reaction CH2I + O2 probed via infrared emission of CO, CO2, OH and H2CO.
The reaction CH2I + O2 has been widely employed recently for the production of the simplest Criegee intermediate CH2OO in laboratories, but the detailed dynamics of this reaction have been little explored. Infrared emission of several products of this reaction, initiated on irradiation of CH2I2 and O2 (∼8 Torr) in a flowing mixture at 308 or 248 nm, was recorded with a step-scan Fourier-transform spectrometer; possible routes of formation were identified according to the observed vibrational distribution of products and published theoretical potential-energy schemes. Upon irradiation at 308 nm, Boltzmann distributions of CO (v ≤ 5, J ≤ 19) with an average vibrational energy of 32 ± 3 kJ mol-1 and OH (v ≤ 3, J ≤ 5.5) with an average vibrational energy of 29 ± 4 kJ mol-1 were observed and assigned to the decomposition of HCOOH* to form CO + H2O and OH + HCO, respectively. The broadband emission of CO2 was simulated with two vibrational distributions of average energies (91 ± 4) and (147 ± 8) kJ mol-1 and assigned to be produced from the decomposition of HCOOH* and methylene bis(oxy), respectively. Upon irradiation of samples at 248 nm, the emission of OH and CO2 showed similar distributions with slightly greater energies, but the distribution of CO (v ≤ 11, J ≤ 19) became bimodal with average vibrational energies of (23 ± 4) and (107 ± 29) kJ mol-1, and branching (56 ± 5) : (44 ± 5). The additional large-v component is assigned to be produced from a secondary reaction HCO + O2 to form CO + HO2; HCO is a coproduct of OH. The branching between CO and OH is (50 ± 5) : (50 ± 5) at 308 nm and (64 ± 5) : (36 ± 4) at 248 nm, consistent with the mechanism according to which an additional channel to produce CO opens at 248 nm. Highly internally excited H2CO was also observed. With O2 at 16 Torr, the extrapolated nascent internal distributions are similar to those with O2 at 8 Torr except for a slight quenching effect.


Quantifying diffusion MRI tractography of the corticospinal tract in brain tumors with deterministic and probabilistic methods.
Diffusion MRI tractography has been increasingly used to delineate white matter pathways in vivo for which the leading clinical application is presurgical mapping of eloquent regions. However, there is rare opportunity to quantify the accuracy or sensitivity of these approaches to delineate white matter fiber pathways in vivo due to the lack of a gold standard. Intraoperative electrical stimulation (IES) provides a gold standard for the location and existence of functional motor pathways that can be used to determine the accuracy and sensitivity of fiber tracking algorithms. In this study we used intraoperative stimulation from brain tumor patients as a gold standard to estimate the sensitivity and accuracy of diffusion tensor MRI (DTI) and q-ball models of diffusion with deterministic and probabilistic fiber tracking algorithms for delineation of motor pathways. We used preoperative high angular resolution diffusion MRI (HARDI) data (55 directions, b = 2000 s/mm(2)) acquired in a clinically feasible time frame from 12 patients who underwent a craniotomy for resection of a cerebral glioma. The corticospinal fiber tracts were delineated with DTI and q-ball models using deterministic and probabilistic algorithms. We used cortical and white matter IES sites as a gold standard for the presence and location of functional motor pathways. Sensitivity was defined as the true positive rate of delineating fiber pathways based on cortical IES stimulation sites. For accuracy and precision of the course of the fiber tracts, we measured the distance between the subcortical stimulation sites and the tractography result. Positive predictive rate of the delineated tracts was assessed by comparison of subcortical IES motor function (upper extremity, lower extremity, face) with the connection of the tractography pathway in the motor cortex. We obtained 21 cortical and 8 subcortical IES sites from intraoperative mapping of motor pathways. Probabilistic q-ball had the best sensitivity (79%) as determined from cortical IES compared to deterministic q-ball (50%), probabilistic DTI (36%), and deterministic DTI (10%). The sensitivity using the q-ball algorithm (65%) was significantly higher than using DTI (23%) (p < 0.001) and the probabilistic algorithms (58%) were more sensitive than deterministic approaches (30%) (p = 0.003). Probabilistic q-ball fiber tracks had the smallest offset to the subcortical stimulation sites. The offsets between diffusion fiber tracks and subcortical IES sites were increased significantly for those cases where the diffusion fiber tracks were visibly thinner than expected. There was perfect concordance between the subcortical IES function (e.g. hand stimulation) and the cortical connection of the nearest diffusion fiber track (e.g. upper extremity cortex). This study highlights the tremendous utility of intraoperative stimulation sites to provide a gold standard from which to evaluate diffusion MRI fiber tracking methods and has provided an object standard for evaluation of different diffusion models and approaches to fiber tracking. The probabilistic q-ball fiber tractography was significantly better than DTI methods in terms of sensitivity and accuracy of the course through the white matter. The commonly used DTI fiber tracking approach was shown to have very poor sensitivity (as low as 10% for deterministic DTI fiber tracking) for delineation of the lateral aspects of the corticospinal tract in our study. Effects of the tumor/edema resulted in significantly larger offsets between the subcortical IES and the preoperative fiber tracks. The provided data show that probabilistic HARDI tractography is the most objective and reproducible analysis but given the small sample and number of stimulation points a generalization about our results should be given with caution. Indeed our results inform the capabilities of preoperative diffusion fiber tracking and indicate that such data should be used carefully when making pre-surgical and intra-operative management decisions.


Sensitive spectrophotometric assay for 3-hydroxy-substituted flavonoids, based on their binding with molybdenum, antimony, or bismuth.
A sensitive spectrophotometric assay has been developed for flavonoids based on their binding with molybdenum, antimony, or bismuth. Acetylation of the hydroxyl group of flavonoids abolished metal binding, thus suggesting a direct role of the hydroxyl groups. From a comparison of several related flavonoids differing in the position of hydroxyl substitutions, the hydroxyl group at position 3 was found to be an important requirement for the formation of a yellow complex. This flavonoid metal complex showed that a specific and significant bathochromic shift in the visible spectrum of the native flavonoid and the corresponding lambda(max) value was used for the colorimetric assays with different metal salts. The molybdenum complex was found to yield higher absorbance compared to antimony and bismuth complexes of various flavonoids. The present method offers a sensitive assay in the 5-25 nM range for these flavonoids and gave comparable results with HPLC quantitative determination.


Use of negative-pressure wound therapy and split-thickness skin autograft to cover an exposed renal transplant.
Exposure of a renal transplant through the abdominal wall is a rare event. A search of the literature reveals only six documented cases which used skin autograft for coverage, with none reported since 1981, and none which used negative-pressure wound therapy (NPWT) to prepare the recipient bed. This case report demonstrates that NPWT followed by split thickness skin graft is a reconstructive option which is feasible in patients who are at high risk for surgical complications in prolonged flap surgery.


STRE- and cAMP-independent transcriptional induction of Saccharomyces cerevisiae GSY2 encoding glycogen synthase during diauxic growth on glucose.
It has been shown that the so-called stationary phase GSY2 gene encoding glycogen synthase was induced as the cells left the exponential phase of growth, while glucose and all other nutrients were still plentiful in the medium (Parrou et al., 1999). Since this effect was essentially controlled at the transcriptional level, we looked for the cis- and trans-acting elements required for this specific growth-related genetic event. We demonstrated that mutations of the HAP2/3/4 binding site and of the two STress-Responsive cis-Elements (STRE) did not abolish the early induction of GSY2, although the latter mutation led to a 20-fold drop in the transcriptional activity of the promoter, as determined from lacZ gene fusions. Insertion of a DNA fragment (from -390 to -167 bp, relative to the ATG) of the promoter lacking the two STREs, upstream to the TATA box of a CYC1-lacZ fusion gene, allowed this reporter gene to be induced with a kinetic similar to that of GSY2-lacZ. Mutations in BCY1, which results in a hyperactive protein kinase A, did not alleviate the early induction, while causing a five- to 10-fold reduction in the transcriptional activity of GSY2. In addition, the repressive effect of protein kinase A was quantitatively conserved when both STREs were mutated in GSY2 promoter, indicating that the negative control of gene expression by the RAS-cAMP signalling pathway does not act solely through STREs. Taken together, these results are indicative of an active process that couples growth control to dynamic glucose consumption.


Natural transformation of Campylobacter jejuni requires components of a type II secretion system.
The human pathogen Campylobacter jejuni is one of more than 40 naturally competent bacterial species able to import macromolecular DNA from the environment and incorporate it into their genomes. However, in C. jejuni little is known about the genes involved in this process. We used random transposon mutagenesis to identify genes that are required for the transformation of this organism. We isolated mutants with insertions in 11 different genes; most of the mutants are affected in the DNA uptake stage of transformation, whereas two mutants are affected in steps subsequent to DNA uptake, such as recombination into the chromosome or in DNA transport across the inner membrane. Several of these genes encode proteins homologous to those involved in type II secretion systems, biogenesis of type IV pili, and competence for natural transformation in gram-positive and gram-negative species. Other genes identified in our screen encode proteins unique to C. jejuni or are homologous to proteins that have not been shown to play a role in the transformation in other bacteria.


Abdominal tuberculosis. A report of 3 cases.
Three patients admitted to the surgical wards of Johannesburg Hospital in whom abdominal tuberculosis was suspected are presented. In every case difficulties were encountered in diagnosis and treatment. Each case presentation is followed by a short commentary on relevant aspects of abdominal tuberculosis.


Epigenetic Aspects of Engineered Nanomaterials: Is the Collateral Damage Inevitable?
The extensive application of engineered nanomaterial (ENM) in various fields increases the possibilities of human exposure, thus imposing a huge risk of nanotoxicity. Hence, there is an urgent need for a detailed risk assessment of these ENMs in response to their toxicological profiling, predominantly in biomedical and biosensor settings. Numerous "toxico-omics" studies have been conducted on ENMs, however, a specific "risk assessment paradigm" dealing with the epigenetic modulations in humans owing to the exposure of these modern-day toxicants has not been defined yet. This review aims to address the critical aspects that are currently preventing the formation of a suitable risk assessment approach for/against ENM exposure and pointing out those researches, which may help to develop and implement effective guidance for nano-risk assessment. Literature relating to physicochemical characterization and toxicological behavior of ENMs were analyzed, and exposure assessment strategies were explored in order to extrapolate opportunities, challenges, and criticisms in the establishment of a baseline for the risk assessment paradigm of ENMs exposure. Various challenges, such as uncertainty in the relation of the physicochemical properties and ENM toxicity, the complexity of the dose-response relationships resulting in difficulty in its extrapolation and measurement of ENM exposure levels emerged as issues in the establishment of a traditional risk assessment. Such an appropriate risk assessment approach will provide adequate estimates of ENM exposure risks and will serve as a guideline for appropriate risk communication and management strategies aiming for the protection and the safety of humans.


Stabilities of avian sarcoma virus RNAs: comparison of subgenomic and genomic species with cellular mRNAs.
The stabilities of B77 avian sarcoma virus intracellular RNAs were compared to the stability of the total cellular poly(A)-containing RNA by labelling infected chicken embryo fibroblasts with [3H]uridine for 15 h, adding actinomycin D (1 microgram per ml) to block further transcription of viral RNA, and selecting virus-specific RNA from the total cellular poly(A)-containing RNA at 3 hourly intervals. The three virus-specific RNA species (9.3, 3.3 and 5.4 kilobases) decayed with half-lives of 7.5, 10, and 15 h, respectively, whereas the bulk of the cellular mRNA decayed with a half-life of 13 h. To correlate these decay rates with the disappearance of mRNA activities, the actinomycin D-treated cells were pulse-labelled with [3H]leucine at 3 hourly intervals after the addition of the drug and virus-specific protein synthesis was assayed by immunoprecipitation. The mRNA activity for the precursor to the non-glycosylated viral structural proteins (Pr76gag) decayed with a half-life of approximately 6 h, whereas the mRNA activity coding for the precursor to the envelope proteins (gPr92env) decayed with a half-life of 14 h. Thus, the rate of decay of the individual mRNA species corresponded reasonably well with the decay rate for the synthesis of two of the corresponding gene products. The results indicated that the 5.4 kb env mRNA is more stable under these conditions than the 9.3 kb gag mRNA but was not significantly more stable than the bulk of the cellular mRNA. Virus particle production following the addition of actinomycin D was determined by the reverse transcriptase assay and by the incorporation of viral genomic 70S RNA into extracellular virions. Both assays yielded similar results and indicated that particle production was inhibited at a rate (t 1/2 = 4 h) somewhat faster than the decay of Pr76gag synthesis or the disappearance of 9.3 kb RNA. It was established by two independent methods (pulse and chase, and approach to isotope equilibrium), however, that the intracellular half-life of the RNA that is packaged into virions is 6 to 7 h. Thus, these results suggest that a single metabolic pool of 9.3 kb RNA exists in avian sarcoma virus-infected cells and is used both as mRNA and as genome RNA.


Ulnar lengthening and shortening.
Alteration in length of the distal ulna may provide an attractive alternative to more destructive procedures commonly used for the treatment of mechanical and degenerative problems at the distal radioulnar joint. Ulnar recession has shown effectiveness in ulnolunate impingement, triangular fibrocartilage tears, and symptomatic ulnar plus variance. It has also been effective in chondromalacia of the ulnar head in the sigmoid notch and in unstable distal radioulnar joints where recession alters the bearing surface and tightens the ulnocarpal ligamentous complex. Ulnar lengthening has been efficacious in relieving the symptoms of Kienböck's disease by decompressing the involved portion of the lunate and distributing joint compressive force on the triquetrum and medial lunate articular surfaces. It may also be of value in some instances of NDCI associated with an ulnar minus variant.


Double-blind, randomized, placebo-controlled clinical trial on the efficacy and tolerability of a physostigmine patch in patients with senile dementia of the Alzheimer type.
Owing to the pharmacokinetic properties of physostigmine when administered by conventional routes, long-term cholinergic treatment of Alzheimer's disease is difficult to manage. In order to overcome the problems associated with the oral and intravenous application of physostigmine, and to improve patients' compliance, a transdermal therapeutic system was developed. The efficacy and tolerability of this system were evaluated in a double-blind, randomized, multicenter study comparing patches containing 30 mg and 60 mg physostigmine with a placebo patch. The clinical trial followed the basic principles of the various guidelines on the evaluation of anti-dementia drugs, and included patients with mild to moderate probable Alzheimer's disease. A total of 204 patients with probable Alzheimer's disease were included in the study. Of these, 136 patients were eligible for the according-to-protocol analysis of efficacy, 167 subjects for the intention-to-treat analysis of efficacy, and 181 patients were included in the safety analysis. In contrast to the hypothesis to be tested, the efficacy of physostigmine was not superior to that of placebo after a treatment period of 24 weeks. On the contrary, there was even a slight, but not statistically significant, trend toward a better outcome in the placebo group. Median physostigmine plasma concentrations of approximately 100 pg/ml were measured, showing a high degree of interindividual variability and no linear dose relationship between the 30 mg and 60 mg dosages. Plasma cholinesterase activity was not significantly affected by physostigmine. The physostigmine patch application in doses of 30 mg and 60 mg apparently did not lead to physostigmine plasma concentrations that were sufficient to compensate for cholinergic deficiencies in affected brain areas and produce clinical benefits. Both the drug and the transdermal system were generally well tolerated under the study conditions. Modifications of the patch system may perhaps make it possible to achieve higher physostigmine plasma concentrations, which seem to be required to induce the expected beneficial effects during long-term treatment of Alzheimer's disease.


Ginsenoside Rc Is a New Selective UGT1A9 Inhibitor in Human Liver Microsomes and Recombinant Human UGT Isoforms.
Ginseng is known to have inhibitory effects on UGT1A9 activity. However, little is known about the inhibitory effects of ginsenosides, the major active compounds in ginseng, on UGT1A9 activity. In vitro investigation of UGT1A9 inhibition by ginsenosides was carried out using human liver microsomes (HLMs). Among 10 ginsenosides, ginsenoside Rc was the strongest inhibitor of UGT1A9-mediated mycophenolic acid glucuronidase activity. Further inhibition kinetic studies using HLMs suggested that ginsenoside Rc competitively and noncompetitively inhibited UGT1A9-mediated propofol and mycophenolic acid glucuronidation activities, with K i values of 2.83 and 3.31 μM, respectively. Next, to investigate whether the inhibitory effect of ginsenoside Rc is specific to the UGT1A9 isoform, we studied the inhibitory potency of ginsenoside Rc on nine human uridine diphospho-glucuronosyltransferase (UGT) activities using recombinant human UGT isoforms. Ginsenoside Rc exhibited a 12.9-fold selectivity (which was similar to niflumic acid at 12.5-fold) for UGT1A9 inhibition. Ginsenoside Rc at 50 μM also inhibited none of the other UGT isoform-specific activities above 12.0%, except for UGT1A9 (>91.5%) in HLMs, indicating that ginsenoside Rc might be used as a selective UGT1A9 inhibitor in reaction phenotyping studies of new chemical entities. Considering lower plasma concentrations (0.01 μM) of ginsenoside Rc in healthy subjects and no induction potential on UGT isoforms, ginsenoside Rc does not cause pharmacokinetic drug interactions with other coadministered drugs metabolized by UGT1A9. SIGNIFICANCE STATEMENT: Ginsenoside Rc selectively inhibited UGT1A9-mediated propofol and mycophenolic acid glucuronidation activities in human liver microsomes and recombinant uridine diphospho-glucuronosyltransferase (UGT) isoforms. It exhibited a 12.9-fold selectivity for UGT1A9 inhibition. Therefore, ginsenoside Rc might be used as a selective UGT1A9 inhibitor in reaction phenotyping studies of new chemical entities, such as niflumic acid.


Effects of intense pulsed light on tissue vascularity and wound healing: a study with mouse island skin flap model.
Intense pulsed light (IPL) has been used extensively in aesthetic and cosmetic dermatology. To test whether IPL could change the tissue vascularity and improve wound healing, mice were separated into 4 groups. Mice in Group I were not treated with IPL, whereas, dorsal skins of mice in Groups II, III, and IV were treated with 35 J/cm(2), 25 J/cm(2), and 15 J/cm(2) IPL, respectively. After 2 weeks, dorsal island skin flaps were raised, based on the left deep circumflex iliac vessels as pedicles; then, survival rate was assessed. Flaps in Group IV (treated with lowest dose of IPL) have a survival rate significantly higher than other groups. Counting blood vessels did not demonstrate any significant differences; however, vessel dilation was found in this group. The results show that IPL at the therapeutic doses which are usually applied to humans is harmful to mouse dorsal skin and did not enhance wound healing, whereas, IPL at much lower dose could improve wound healing. The possible mechanism is the dilation of tissue vasculature thanks to the electromagnetic character of IPL. Another mechanism could be the heat-shock protein production.


The diagnosis of fetal hydrocephalus before viability.
A diagnosis of fetal hydrocephalus was made before viability. The diagnosis was indicated by a discrepancy between the sonographic fetal head size, the uterine size by both clinical and sonographic examination, and x-ray films. An abortion was performed by hysterotomy.


[A proposed open formulary for a managed care program in Mexico. The Therapeutic and Pharmacological Committee of Mexico].
To integrate an open formulary with drug use recommendations in which a physician could find a wide range of therapeutic options for his private practice. An interdisciplinary medical committee with a high scientific level, reviewed each one of all the drugs available to be sell in Mexico, as part of a Pharmacy Benefit Management & Manage Care program implemented in our country. Each drug was classified according to its active ingredient under one of the following categories: 1) essentials, 2) excluded, and 3) not essentials-not excluded. The decision in each case was based on pure scientific grounds and using an Evidence-Based Medicine analysis. The proposed formulary contains a total of 1106 active ingredients-either unique or in combination-, from which 429 (38.8%) were classified as essentials. About one-out-of-ten drugs (8.7%) currently in use within the private medical practice in Mexico, some of them highly prescribed, were regarded by the committee as excluded due to concerns about their efficacy and/or safety. On this report a detailed description of the process followed to conform the committee, its objectives an its performing criteria is presented, as well as the bases under which the final list of drugs of the formulary were set.


Vibration-resistant phase-shifting interferometry.
A method to reduce the sensitivity of phase-shifting interferometry to external vibrations is described. The returning interferogram is amplitude split to form two series of interferograms, taken simultaneously and with complementary properties, one with high temporal and low spatial resolution and the other with low temporal and high spatial resolution. The high-temporal-resolution data set is used to calculate the true phase increment between interferograms in the high-spatial-resolution data set, and a generalized phase-extraction algorithm then includes these phase increments when the topographical phases in the high-spatial-resolution data set are calculated. The measured topography thereby benefits from the best qualities of both data sets, providing increased vibration immunity without sacrificing high spatial resolution.


GRS1, a yeast tRNA synthetase with a role in mRNA 3' end formation.
Transcription termination and 3' end formation are essential processes necessary for gene expression. However, the specific mechanisms responsible for these events remain elusive. A screen designed to identify trans-acting factors involved in these mechanisms in Saccharomyces cerevisiae identified a temperature-sensitive mutant that displayed phenotypes consistent with a role in transcription termination. The complementing gene was identified as GRS1, which encodes the S. cerevisiae glycyl-tRNA synthetase. This result, although unusual, is not unprecedented given that the involvement of tRNA synthetases in a variety of cellular processes other than translation has been well established. A direct role for the synthetase in transcription termination was determined through several in vitro assays using purified wild type and mutant enzyme. First, binding to two well characterized yeast mRNA 3' ends was demonstrated by cross-linking studies. In addition, it was found that all three substrates compete with each other for binding to GlyRS enzyme. Next, the affinity of the synthetase for the two mRNA 3' ends was found to be similar to that of its "natural" substrate, glycine tRNA in a nitrocellulose filter binding assay. The effect of the grs1-1 mutation was also examined and found to significantly reduce the affinity of the enzyme for the three RNA substrates. Taken together, these data indicate that not only does this synthetase interact with several different RNA substrates, but also that these substrates bind to the same site. These results establish a direct role for GRS1 in mRNA 3' end formation.


Ex vivo and in vivo adenovirus-mediated gene therapy strategies induce a systemic anti-tumor immune defence in the B16 melanoma model.
The efficacy of adenovirus-mediated gene therapy for treatment of metastatic B16 melanomas, established in syngeneic C57BL/6 mice, was assessed via an ex vivo cytokine vaccine approach or via an in vivo strategy utilizing combination cytokine/herpes simplex virus-thymidine kinase (HSV-tk) suicide gene delivery and treatment with ganciclovir (GCV). In the ex vivo tumor vaccine approach, B16 melanoma cells, transduced in vitro by adenovirus containing either interleukin (IL)-2, granulocyte-macrophage colony stimulating factor (GM-CSF), or tumor necrosis factor-alpha cytokine genes and gamma irradiated, were subcutaneously injected into the flank and a distant subcutaneous challenge injection of unmodified B16 melanoma cells was performed 15 d later. Significant reductions in challenge tumor volume were observed in the IL-2 group (75% reduction; p = 0.02) and in the GM-CSF group (88% reduction; p = 0.0006), whereas the effect for tumor necrosis factor-alpha was not statistically significant. In the in vivo treatment of established melanomas, this cytokine approach was combined with a suicide gene therapy and subcutaneous B16 melanomas were directly injected with (i) IL-2/recombinant, replication-deficient adenovirus (adv) and thymidine kinase (tk)/adv, (ii) GM-CSF/adv, IL-2/adv, and tk/adv, or (iii) control beta-galactosidase (beta-gal)/adv and tk/adv. After intraperitoneal application of GCV (10 mg per kg) for 6 d, the residual tumor masses were excised and the animals challenged with unmodified B16 cells. Challenge tumor growth was reduced by 56% for the IL-2/tk/adv/GCV treatment (p = 0.041) and by 77% for the GM-CSF/IL-2/tk/adv/GCV treatment p (p = 0.037), in comparison with the beta-gal/tk/GCV control group. These data may hold significant promise for the development of effective ex vivo and in vivo gene therapy modalities to counter the highly metastatic nature of human melanoma.


Prosthesis infections after orthopedic joint replacement: the possible role of bacterial biofilms.
Prosthesis-related infection is a serious complication for patients after orthopedic joint replacement, which is currently difficult to treat with antibiotic therapy. Consequently, in most cases, removal of the infected prosthesis is the only solution to cure the infection. It is, therefore, important to understand the comprehensive interaction between the microbiological situation and the host immune responses that lead to prosthesis infections. Evidence indicates that prosthesis infections are actually biofilm-correlated infections that are highly resistant to antibiotic treatment and the host immune responses. The authors reviewed the related literature in the context of their clinical experience, and discussed the possible etiology and mechanism leading to the infections, especially problems related to bacterial biofilm, and prophylaxis and treatment of infection, including both microbiological and surgical measures. Recent progress in research into bacterial biofilm and possible future treatment options of prosthesis-related infections are discussed.


Mouse models of DNA mismatch repair in cancer research.
Germline mutations in DNA mismatch repair (MMR) genes are the cause of hereditary non-polyposis colorectal cancer/Lynch syndrome (HNPCC/LS) one of the most common cancer predisposition syndromes, and defects in MMR are also prevalent in sporadic colorectal cancers. In the past, the generation and analysis of mouse lines with knockout mutations in all of the known MMR genes has provided insight into how loss of individual MMR genes affects genome stability and contributes to cancer susceptibility. These studies also revealed essential functions for some of the MMR genes in B cell maturation and fertility. In this review, we will provide a brief overview of the cancer predisposition phenotypes of recently developed mouse models with targeted mutations in MutS and MutL homologs (Msh and Mlh, respectively) and their utility as preclinical models. The focus will be on mouse lines with conditional MMR mutations that have allowed more accurate modeling of human cancer syndromes in mice and that together with new technologies in gene targeting, hold great promise for the analysis of MMR-deficient intestinal tumors and other cancers which will drive the development of preventive and therapeutic treatment strategies.


Introducing novel electronic and magnetic properties in C3N nanosheets by defect engineering and atom substitution.
Using first-principles calculations the effect of topological defects, vacancies, Stone-Wales and anti-site and substitution of atoms, on the structure and electronic properties of monolayer C3N are investigated. Vacancy defects introduce localized states near the Fermi level and a local magnetic moment. While pristine C3N is an indirect semiconductor with a 0.4 eV band gap, with substitution of O, S and Si atoms for C, it remains a semiconductor with a band gap in the range 0.25-0.75 eV, while it turns into a metal with H, Cl, B, P, Li, Na, K, Be and Mg substitution. With F substitution, it becomes a dilute-magnetic semiconductor, while with Ca substitution it is a ferromagnetic-metal. When replacing the N host atom, C3N turns into: a metal (H, O, S, C, Si, P, Li and Be), ferromagnetic-metal (Mg), half-metal (Ca) and spin-glass semiconductor (Na and K). Moreover, the effects of charging and strain on the electronic properties of Na atom substitution in C3N are investigated. We found that the magnetic moment decreases or increases depending on the type and size of strain (tensile or compression). Our study shows how the band gap and magnetism in monolayer C3N can be tuned by introducing defects and atom substitution. The so engineered C3N can be a good candidate for future low dimensional devices.


Periodontal microsurgery: report of 16 cases consecutively treated by the free rotated papilla autograft technique combined with the coronally advanced flap.
The coronally advanced flap combined with a free connective tissue graft is a predictable method for achieving root coverage in buccal gingival recession. Nevertheless, this procedure conventionally requires involvement of a second surgical site; the latter is avoided by the proposed technique. Sixteen isolated gingival recessions (2.5 to 4.0 mm deep) were surgically treated with a coronally advanced flap associated with a connective tissue graft harvested from one adjacent papilla whose dimensions matched those of the exposed root area. Procedures were performed with the aid of a surgical microscope. Recession depth, probing depth, periodontal attachment level, and keratinized tissue width were recorded at baseline and 12 months after surgery. Mean recession moved from 3.38 +/- 0.72 mm at baseline to 0.13 +/- 0.29 mm at 12 months, a gain of 97.03%. In 13 of the 16 cases 12 months after surgery, the gingival margin was located at the CEJ or coronal to it, while in two cases the residual recession was less than 1.0 mm and in another case it was 1.0 mm. Mean periodontal attachment level was 4.72 +/- 1.00 mm at baseline and 1.03 +/- 0.59 mm at follow-up. Mean keratinized tissue increased from 1.25 +/- 0.75 mm to 3.47 +/- 0.87 mm. All differences between 12 months and baseline were statistically significant. No pockets were present at baseline, and this situation remained stable during the observation period. All 16 isolated recessions treated showed an excellent gain in root coverage without requiring a second surgical site and thus reducing patient morbidity.


Usefulness of sleep apnea monitoring by pacemaker sensor in elderly patients with diastolic dysfunction.
Automated detection of sleep apnea (SA) by pacemaker (PM) has been proposed and exhibited good agreement with polysomnography to detect severe SA. We aimed to evaluate the usefulness of SA monitoring algorithm in elderly patients with diastolic dysfunction. Consecutive patients referred to the Caen University Hospital for PM implantation between May 2016 and December 2018 presenting isolated diastolic dysfunction were eligible for the study. The respiratory disturbance index (RDI) measured by the PM, and the mean monthly RDI (RDIm), were compared to the apnea hypopnea index (AHI) assessed with portable monitor for severe SA diagnosis. During the study period, 68 patients were recruited, aged of 80.4 ± 8.2 years. 63 patients underwent polygraphy with a portable monitor: 57 presented SA (83.8%), including 16 with severe SA (23.5%). Eight were treated with continuous positive airway pressure (CPAP). We found the RDI cutoff value of 22 events/h to predict severe SA, with 71.4% sensitivity and 65.2%, specificity. The RDIm cutoff value to detect severe SA was 19 events/h, with a sensitivity of 60% and a specificity of 66%. There was a significant reduction in RDI (p = 0.041), RDIm (p = 0.039) and AHI (p = 0.002) after CPAP. Supraventricular arrhythmias were frequent in all patients, regardless of SA severity, considering either episodes occurrence or total burden. In a population of elderly patients with PM and diastolic dysfunction, the SA monitoring algorithm was able to detect severe SA, with good diagnostic performance values, but also to provide follow-up data for the patients treated with CPAP.


Amphotericin B colloidal dispersion.
Amphotericin B colloidal dispersion (ABCD) is a colloidal dispersion of a stable complex of amphotericin B with cholesteryl sulphate in a 1:1 proportion, forming uniform disk-shaped particles. ABCD is associated with less nephrotoxicity than conventional amphotericin B deoxycholate. Infusion-related adverse events are more frequent in patients receiving ABCD than in patients receiving liposomal amphotericin B or amphotericin B deoxycholate. ABCD has been shown in a randomised, double-blind study, to be an effective alternative to amphotericin B deoxycholate for empirical treatment of patients with fever and neutropenia. ABCD is active in the treatment of invasive Candida spp. and Aspergillus spp. infections in immunocompromised hosts, however most of the data supporting its use for these types of infections is derived from non-comparative open-label clinical trials of patient refractory to or intolerant of conventional antifungal therapy. ABCD is approved by the US FDA for the treatment of invasive aspergillosis in patients where renal impairment of unacceptable toxicity precludes the use of amphotericin B deoxycholate in effective doses, and in patients with invasive aspergillosis where prior amphotericin B deoxycholate therapy has failed. Two other lipid formulations of amphotericin B, amphotericin B lipid complex and liposomal amphotericin B, are available and, like ABCD, are associated with reduced nephrotoxicity as compared to amphotericin B deoxycholate. The role of ABCD in comparison with these other lipid formulations of amphotericin B is discussed herein. High cost remains an issue with all lipid formulations of amphotericin B.


Comparison of the lower confidence limit to the fixed-percentage method for assessing airway obstruction in routine clinical practice.
Although the statistically derived lower limit of normal (LLN) for the ratio of FEV(1) to FVC is considered superior to a fixed cutoff value (such as 0.70) for diagnosing airway obstruction, the fixed-cutoff method continues to be used and advocated. To evaluate the misclassification of spirometrically determined airway obstruction arising from the use of the fixed-percent method, in comparison to the LLN method for FEV(1)/FVC. We reviewed 27,307 spirometry records from adult men, and diagnosed airway obstruction based on the LLN (predicted value minus 1.645 times the standard error of estimate from a north Indian reference equation for FEV(1)/FVC) and based on a fixed cutoff of 0.70. We computed agreement and discordance between the two methods, and determined the sensitivity, specificity, and predictive values of the fixed-percent method in identifying true obstruction. The results were discordant in 1,622 subjects (6%). Overall agreement between the two methods was good (kappa estimate 0.869), but worsened considerably with advancing age. 1,290 subjects (5%) who were deemed normal with the LLN method were diagnosed as having airway obstruction with the fixed-percentage method. Overall the sensitivity, specificity, and positive predictive value of the fixed-percentage method were 0.963, 0.929, and 0.871, respectively. Specificity and positive predictive value decreased sharply with advancing age. The negative age-dependence of FEV(1)/FVC results in over-diagnosis of airway obstruction in middle-aged and elderly men, and under-diagnosis in young men, with the fixed-percentage method. Airway obstruction should be assessed with the LLN of FEV(1)/FVC, with the LLN derived from appropriate reference equations.


Soft tissue calcification in the Ossabaw miniature pig: experimental and kinetic modeling studies.
Calcium (Ca) deposition into vascular tissue was measured in Ossabaw miniature pigs with and without metabolic syndrome (MetS) using Ca tracer kinetics and coronary atherosclerosis measured with intravascular ultrasound. Pigs with MetS had higher Ca uptake into coronary arteries than lean pigs. Ca deposition into arteries is a common disease in humans. The Ossabaw pig develops MetS when fed an atherogenic diet. The aim of this study was to measure Ca deposition into arteries of lean vs. MetS pigs. Male pigs were fed for 5 months with chow diet (healthy, lean; n = 7) or atherogenic diet (n = 8) consisting of chow supplemented with 2 % cholesterol, 43 % kcal from fat, and 20 % kcal from fructose. Pigs were verified to have MetS by obesity, insulin resistance, impaired glucose tolerance, dyslipidemia, and hypertension. Two pigs received 50 nCi of (41)Ca i.v. and blood was drawn frequently for 24 h, and 2, 3, 6, 8, 10, 15, 20, and at sacrifice at 28 days after injection. Peripheral arteries were biopsied four times per pig over the 28th day and coronary artery sampled at sacrifice. Tissues were analyzed for (41)Ca:Ca. A compartmental model was used to estimate rates of Ca deposition into the arteries. The MetS swine had higher (41)Ca and atherosclerosis in coronary arteries than lean pigs. This pig model is a suitable model for studying vascular calcification in humans.


Efficacy of a single oral dose of 200,000 IU of oil-soluble vitamin A in measles-associated morbidity.
The authors assessed the efficacy of the World Health Organization (WHO) recommendation of 200,000 IU of vitamin A in oil to treat acute non-xerophthalmic measles patients. Acute measles patients who did not require hospitalization were enrolled in a randomized, double-masked, clinical trial of vitamin A (n=90) versus placebo (n=110) carried out in Ndola, Zambia, in 1991. Measles-associated morbidity was defined by the presence of signs and symptoms of acute respiratory infection. Daily evaluations for the first 3 days were followed by weekly visits for a month at urban health centers. Baseline demographic, clinical, and biochemical characteristics were similar in both groups. Cross-sectional analysis of morbidity status, by group, at each weekly evaluation showed no significant differences until week 4, when more placebo-treated patients had cough or pneumonia (p=0.005). However, longitudinal analysis, which looked at changes among individuals and controlled for initial health status, showed more equivocal results. The odds ratio for the development of pneumonia in patients with measles cough in vitamin A-treated subjects was 0.73 (95% confidence interval (Cl) 0.30-1.80). The odds ratio for the development of measles-associated cough or pneumonia in asymptomatic measles patients was 0.52 (95% Cl 0.24-1.13), in favor of vitamin A, but the odds ratio for failing to improve from pneumonia in vitamin A-treated subjects was 1.23 (95% Cl 0.68-2.3), a result in favor of placebo. These results suggest that the evidence for the efficacy of one dose of vitamin A in oil to prevent measles complications is not as strong as that previously shown for two 200,000 IU doses of water-miscible vitamin A, and that the WHO recommendation may need to be reexamined.


Multiple doublesex-related genes specify critical cell fates in a C. elegans male neural circuit.
In most animal species, males and females exhibit differences in behavior and morphology that relate to their respective roles in reproduction. DM (Doublesex/MAB-3) domain transcription factors are phylogenetically conserved regulators of sexual development. They are thought to establish sexual traits by sex-specifically modifying the activity of general developmental programs. However, there are few examples where the details of these interactions are known, particularly in the nervous system. In this study, we show that two C. elegans DM domain genes, dmd-3 and mab-23, regulate sensory and muscle cell development in a male neural circuit required for mating. Using genetic approaches, we show that in the circuit sensory neurons, dmd-3 and mab-23 establish the correct pattern of dopaminergic (DA) and cholinergic (ACh) fate. We find that the ETS-domain transcription factor gene ast-1, a non-sex-specific, phylogenetically conserved activator of dopamine biosynthesis gene transcription, is broadly expressed in the circuit sensory neuron population. However, dmd-3 and mab-23 repress its activity in most cells, promoting ACh fate instead. A subset of neurons, preferentially exposed to a TGF-beta ligand, escape this repression because signal transduction pathway activity in these cells blocks dmd-3/mab-23 function, allowing DA fate to be established. Through optogenetic and pharmacological approaches, we show that the sensory and muscle cell characteristics controlled by dmd-3 and mab-23 are crucial for circuit function. In the C. elegans male, DM domain genes dmd-3 and mab-23 regulate expression of cell sub-type characteristics that are critical for mating success. In particular, these factors limit the number of DA neurons in the male nervous system by sex-specifically regulating a phylogenetically conserved dopamine biosynthesis gene transcription factor. Homologous interactions between vertebrate counterparts could regulate sex differences in neuron sub-type populations in the brain.


Hepatitis C: a changing epidemic.
Approximately 3% of the world population is estimated to have a chronic hepatitis C virus (HCV) infection and 500,000 individuals die from its consequences yearly. Persons who inject drugs (PWID) bear the majority of the disease burden in high-income countries. Drug substitution programmes have helped reduce HCV transmissions among PWID. However, recent epidemics of sexually transmitted HCV infections in HIV-infected men who have sex with men demonstrated the changing nature of the HCV epidemic. HCV therapy is undergoing a revolution, as new interferon-free, oral treatments eradicate HCV infections in almost all treated patients. As a consequence, the eradication of HCV has become a matter of debate and is becoming an important future public health target. However, for this to be achieved, many challenges need to be addressed, including the poor uptake of HCV testing, the high cost of the new antiviral combinations and the high frequency of re-infections after treatment in some populations.


Hospital teaching status and patient outcomes in intestinal obstruction surgery: A comparative analysis.
Surgery at large teaching hospitals is reportedly associated with more favourable outcomes. However, these results are not uniformly consistent across all surgical patients. This study aimed to assess potential disparities in clinical outcomes by hospital type for patients with intestinal obstruction. 2018 NIS was queried for all adult non-elective admissions for intestinal obstruction. Hospitals were classified as either smallmedium non-teaching hospitals or large teaching hospitals. Multivariate regression analyses were used to assess the association between hospital type and inpatient mortality, access to surgery, admission duration, non-home discharges, hospital costs, and postoperative complications. After adjustments, admission to large teaching hospitals was not associated with a reduction in inpatient mortality (AOR= 0.73; 95% CI= 0.41- 1.31; p= 0.29), lower likelihood of surgery (AOR= 0.93; 95% CI= 0.58-1.48; p= 0.76) or increased chance of early surgery (p= 0.97). Patients admitted to large teaching hospitals had shorter hospital stays (p= 0.002) and were less likely to be discharged to other acute care hospitals (AOR= 0.94; 95% CI= 0.80-0.94; p= 0.04). Admission to large teaching hospitals was not associated with a reduction in perioperative complications (AOR= 1.04; 95% CI= 0.80- 1.28; p= 0.91) or significantly higher hospital costs (mean increase= 1518; 95% CI= 1891-4927; p= 0.38). Admission to large teaching hospitals does not necessarily result in better patient outcomes. Merely considering the teaching status of the hospital in isolation cannot explain the diverse outcomes observed for this condition.