Comprehensive clinical characterization of patients with BRCA1: c.5017_5019del germline variant.
We provide evidence for the reclassification of the BRCA1:c.5017_5019del variant by presenting the clinicopathological characteristics, clinical outcomes, and family history of breast or ovarian cancer in 17 patients with this variant. This study included breast or ovarian cancer patients tested for BRCA1/2 genes between January 2008 and June 2020 at 10 medical centers in Korea. We retrospectively reviewed 17 probands from 15 families who had the BRCA1:c.5017_5019del variant according to the electronic medical records. We present 10 breast cancer patients and 7 ovarian cancer patients from 15 families identified as having BRCA1:c.5017_5019del and a total of 19 cases of breast cancer and 14 cases of ovarian cancer in these families. The ratio of breast-to-ovarian cancer was 1.3:1. Breast cancer patients with this variant showed a rich family history of breast or ovarian cancer, 8 patients (80.0%). The mean age at diagnosis was 45.4 years and 6 patients (60.0%) were categorized into hormone-receptor-negative breast cancer. Also, the ovarian cancer patients with this variant showed strong family histories of breast and/or ovarian cancer in 4 patients (57.1%). We presented clinical evidence for the reclassification of BRCA1:c.5017_5019del as a likely pathogenic variant (LPV). Reclassification as LPV could result in the prophylactic treatment and medical surveillance of probands, family testing recommendations, and appropriate genetic counseling of their families.


Selection of primers of optimal sensitivity for the detection of HIV-1 from Africa and Europe by polymerase chain reaction.
In order to facilitate the detection of integrated HIV-1 proviral DNA from African as well as European patients, four new primer pairs for use in the polymerase chain reaction (PCR), localised in the gag, pol, vif and env genes of HIV-1, were constructed. The primer pairs were compared to all accessible HIV-1 sequences from African and European isolates and to some of the earlier published and most commonly used primer pairs. HIV-1 DNA was detected in blood drawn from 13 out of 13 individuals infected in Africa, in three out of three Tanzanian HIV-1 isolates and in three out of three asymptomatic Swedes infected in Europe. The new selection of primer pairs can be used as an alternative to enhance the detection of HIV-1 of different origins.


Cadaver-donor renal retransplants.
1. There was a dramatic improvement in survival rates for retransplanted patients between 1989 and 1991. The proportion of cadaveric kidneys used to retransplant patients has decreased from 18.4% in 1984 to 14.3% in 1992. In 1992, the 81% one-year survival rate for second transplants nearly reached that of first transplants. 2. Despite recent improvements in regraft survival, the PGST remained the strongest predictor of second graft outcome. One-year regraft survival rates were 54% when the PGST was one to 3 months and 75% when the PGST was more than one year. The steady improvement in second graft survival rates may be influenced by retransplanting more patients with a longer PGST. 3. Patients with broadly reactive anti-HLA antibodies (PRA > 50%) had 10% lower regraft survival rates than nonsensitized patients (p < 0.001). 4. A repeated HLA-DR antigen mismatch resulted in 6% and 13% lower regraft survival at one and 3 years, respectively, compared with patients who had at least one HLA-DR mismatch, but not for the same antigen mismatched previously (p < 0.05). 5. Recipients expressing HLA-DR1 had an 81% one-year second graft survival rate compared with 75% for patients lacking DR1 (p < 0.001). 6. Matching for HLA-B,DR and HLA-A,B,DR antigen combinations resulted in 80-82% one-year regraft survival rates for second and multiple transplant recipients, compared with 57-74% for completely mismatched transplants. 7. Transplants from donors under age 6 or over age 55 had significantly poorer outcomes than those involving kidneys from donors in the intermediate age range. The results of transplants at the extremes of donor age were very poor when the recipient was retransplanted. 8. Among recipients of second and multiple transplants, prophylactic OKT3 yielded 82% and 72% one-year regraft survival, respectively, compared with 75% and 66% with no antibody induction (p < 0.001 for second transplants). 9. Regraft survival rates comparable with those for primary cadaveric transplants reported to the UNOS Registry since 1991 justify the use of cadaver-donor kidneys for low-risk patients seeking second renal transplants.


Evaluation of a simulation-based program for medic cognitive skills training.
Simulation-based training is a promising instructional approach for training military and civilian first responders. In addition to training in relevant taskwork skills, there is increasing need for first responder training in cognitively-based skills such as situation assessment and decision making. The First Responder Simulation Training (FIRST) program trains cognitive skills using complex and degraded situations. The program is comprised of five detailed scenarios, evaluation instruments, debriefing guidelines for each scenario, a multimedia tutorial that explains how to use the evaluation and debriefing instruments, and a detailed scenario guide for administering the scenarios. We conducted an evaluation of the FIRST program to assess its training utility and usability. The program was well-received by both instructors and participants. Instructors noted the importance of training cognitive skills and found the instructor materials valuable for teaching them how to administer a simulation-based training program. Participants found the scenarios realistic and challenging, and noted that such simulation-based training would be a valuable supplement to medic curricula.


Heat shock proteins as vaccine candidates.
Parasite heat shock proteins are phylogenetically well-conserved antigenic mosaics that nonetheless stimulate both humoral and cellular immune responses during the course of infection. These range from highly specific to broadly cross-reactive, with intermediate degrees also documented. The latter may prove beneficial, if they can be appropriately channelled to protect against multiple pathogens, or potentially harmful, if they cross-react with host components. Assessment of heat shock proteins as vaccine candidates has thus proceeded cautiously, with efforts being made to map the specificity of host immune responses to individual epitopes of the molecules. Ultimately, to serve as vaccines, the molecules must be recognized by the immune system within the context of a living pathogen, and be capable of inducing appropriate cellular and/or humoral immune responses that are effective at preventing establishment of individual pathogens. Reviewed are studies relevant to the use of parasite hsps as vaccine components, with emphasis on those from schistosomes, malaria, chlamydial, and mycobacterial parasites.


Disparities in defining disparities: statistical conceptual frameworks.
Motivated by the need to meaningfully implement the Institute of Medicine's (IOM's) definition of health care disparity, this paper proposes statistical frameworks that lay out explicitly the needed causal assumptions for defining disparity measures. Our key emphasis is that a scientifically defensible disparity measure must take into account the direction of the causal relationship between allowable covariates that are not considered to be contributors to disparity and non-allowable covariates that are considered to be contributors to disparity, to avoid flawed disparity measures based on implausible populations that are not relevant for clinical or policy decisions. However, these causal relationships are usually unknown and undetectable from observed data. Consequently, we must make strong causal assumptions in order to proceed. Two frameworks are proposed in this paper, one is the conditional disparity framework under the assumption that allowable covariates impact non-allowable covariates but not vice versa. The other is the marginal disparity framework under the assumption that non-allowable covariates impact allowable ones but not vice versa. We establish theoretical conditions under which the two disparity measures are the same and present a theoretical example showing that the difference between the two disparity measures can be arbitrarily large. Using data from the Collaborative Psychiatric Epidemiology Survey, we also provide an example where the conditional disparity is misled by Simpson's paradox, whereas the marginal disparity approach handles it correctly.


Transcriptional regulation by thyroid hormone of an mRNA homologous to a protease inhibitor.
We have previously cloned a cDNA of a rat liver mRNA, designated 4-12B, markedly induced by triiodothyronine (T3) at a pretranslational level [Magnuson, M.A., Dozin, B., & Nikodem, V.M. (1985) J. Biol. Chem. 260, 5906-5912]. Here we show that this hormonal effect is due in part to an increase of the rate of transcription of the 4-12B gene. In addition, the nucleotide sequence of 4-12B cDNA has been determined, revealing significant similarity with the sequences of the superfamily of serine protease inhibitors and a very high homology with contrapsin, a mouse serum trypsin inhibitor, at the level of nucleotide and amino acid sequence (77.9 and 66.8%, respectively). The optimized alignment of the putative reactive center region of 4-12B with four related members of this superfamily revealed that lysine-serine residues are located at the reactive site or adjacent to it, thus suggesting that the triiodothyronine-regulated rat 4-12B mRNA might code for a protease inhibitor with trypsin-like specificity. Although not enough data are presently available to assign definitively antitryptic activity to this protein, the high degree of similarity with members of the superfamily of serine protease inhibitors leaves no doubt that 4-12B is a member of this superfamily.


Exchange of surface proteins impacts on viral vector cellular specificity and transduction characteristics: the retina as a model.
Recombinant vectors based on adeno-associated virus (AAV) or human immunodeficiency 1 (lentivirus) are promising tools for long term in vivo gene delivery. Their design allows the exchange of capsids or envelopes, respectively, theoretically providing the opportunity to transduce a range of cell types. We constructed AAV vectors encoding enhanced green fluorescent protein (EGFP) within an AAV serotype 2 (AAV2) genome contained in an AAV2, five or one capsid (called AAV2/2, AAV2/5 and AAV2/1, respectively). Similarly we produced lentiviral vectors, encoding the same expression cassette present in the AAV vectors, pseudotyped with proteins from vesicular stomatitis virus glycoprotein (VSVG) or Mokola envelopes. Transduction characteristics of these vectors were evaluated in the murine retina following subretinal or intravitreal administration. The time of onset of transgene expression and the targeted cell types differed between the various recombinants. Onset of transgene expression was 3-4 days for lentiviral vectors and AAV2/1. In contrast, onset was at 2-4 weeks for AAV2/5 and AAV2/2, respectively. After subretinal injection, both lenti-VSVG and AAV2/5 transduced the retinal pigment epithelium (RPE) and photoreceptors efficiently whereas transgene expression was restricted to RPE cells using lenti with the Mokola envelope or AAV2/1. After intravitreal administration, only AAV2/2 and lenti-VSVG transduced the inner retina. Vector-mediated fluorescence was detected in the retina for over 12 weeks for all of the vectors. We conclude that pseudotyping provides a useful means to manipulate viral vector cell targeting specificity as well as retinal transduction characteristics of vectors containing the same genome.


Psoriasis and physical activity: a review.
Psoriasis is a common, chronic inflammatory skin disease that can cause significant discomfort and impairment to quality of life. Recent research indicates that individuals with moderate-to-severe psoriasis are likely at greater risk for chronic cardiometabolic co-morbidities such as cardiovascular disease, type 2 diabetes, obesity and metabolic syndrome. Physical activity can be an effective primary and adjunctive treatment for these maladies in other populations. Unfortunately, only a limited number of studies have examined physical activity in psoriasis, which are limited by poor design and lack of validated physical activity assessment methodologies. A variety of data suggest shared physiologic pathways between physical activity, psoriasis, and psoriasis cardiometabolic co-morbidities. Increased adiposity, inflammation, oxidative stress, adhesion molecules and lipids are physiologically linked to psoriasis, the risk of psoriasis cardiometabolic co-morbidities, and low levels of physical activity. In addition, epigenetic pathways are involved in psoriasis and could be influenced by physical activity. The physical and psychosocial impairments common in psoriasis may make it difficult to participate in regular physical activity, and future studies should aim to determine if physical activity interventions improve functioning and reduce co-morbidities in psoriasis.


First Detection of Ligustrum necrotic ringspot virus, Cucumber mosaic virus, and Alternanthera mosaic virus in Mazus reptans in the United States.
Mazus reptans N.E. Br (creeping mazus; Phrymaceae) is a perennial flowering groundcover plant. A plant of M. reptans 'Alba' with mild mosaic symptoms was obtained from a Maryland nursery in 2010. Electron microscopy (EM) revealed slightly flexuous particles of 595 to 674 nm in length and smaller fragments, typical of carlaviruses. This sample was analyzed using a recently-developed Universal Plant Virus Microarray (UPVM [4]), and UPVM results confirmed by RT-PCR and sequencing. For UPVM analysis, complementary DNA (cDNA) was prepared from total nucleic acid extracts using a combination of oligo(dT) and random (6- to 9-mer) primers and high copy sequences (primarily ribosomal) were reduced using duplex-specific nuclease. Treated cDNA was labeled by incorporation of amino-allyl dUTP, followed by coupling of Cy3 dye and hybridization to a UPVM slide (4). Analysis of UPVM hybridization results using associated Uchip and T-Predict software (4) identified Ligustrum necrotic ringspot virus (LNRV; Carlavirus) and Cucumber mosaic virus subgroup I (CMV sgI; Cucumovirus). To confirm the UPVM results, we used NSNC-odT (3) primed cDNA, and LNRV-specific primer Lig1 (GTTGATCCTTTAGGTTTACAGGT) paired with NSNC-odT to amplify the 3' region of the LNRV genome. We used random-primed cDNA with generic cucumovirus coat protein (CP) primers CPTALL-5/CPTALL-3 (2), and CMV subgroup (sg)-specific primers CMV I(F)/CMV I(R) and CMV II(F)/CMV II(R) (1) to amplify the full CMV CP gene or internal portions. A ~1.35 kb PCR product from the LNRV-specific amplification was cloned, sequenced (GenBank Accession No. KJ187250), and found to have 84.6% nt identity to the LNRV-type (EU074853), with 97.0% CP amino acid (AA) identity and 94.7% nucleic acid binding protein (NABP) AA identity to LNRV-Impatiens (GQ411367) excluding an additional 14 N-terminal AA present in the NABP of both the type and impatiens isolates. CMV sgI-specific primers yielded a product of ~600 bp, and generic primers CPTALL-5/CPTALL-3 a ~940 bp product; no product was obtained with sgII-specific primers. The full CP gene product was cloned and sequenced (KJ486271), and had 99% nt identity to CMV-Fny (U20668), a subgroup I isolate, and <75% to characterized sgII isolates (5); CMV-Mazus CP had 100% AA identity to CMV-Fny, and <82.6% to the sgII isolates. One plant of purple M. reptans obtained in 2012, and four purple-flowered and three 'Alba' in 2014 from three separate sources, also showed mild mosaic. LNRV was detected by EM of carlavirus-like particles (2012 sample), and in all eight plants by LNRV-specific PCR and sequencing (KJ187247 for 2012 sample). Alternanthera mosaic virus (AltMV; Potexvirus) was also detected from two plants of 'Alba' by PCR, sequencing, bioassay (Nicotiana benthamiana, Chenopodium quinoa), and ELISA (3). To our knowledge, this is the first report of LNRV, CMV, or AltMV in M. reptans, a commonly grown groundcover plant. While CMV and AltMV are known to have wide host ranges, LNRV has previously been reported only from Ligustrum and Impatiens sp. The mild symptoms hinder symptom-based detection, and M. reptans may thus serve as a conduit for LNRV, CMV, and AltMV infection of other ornamentals. References: (1) S. Chen et al. Acta Biochim. Biophys. Sin. 43:465, 2011. (2) S. K. Choi et al. J. Virol. Meth. 83:67, 1999. (3) J. Hammond et al. Arch. Virol. 151:477, 2006. (4) J. Hammond et al. Phytopathology 102(S4):49, 2012. (5) J. Thompson and M. Tepfer. J. Gen. Virol. 90:2293, 2009.


COVID-19 vaccination-related autoimmune hepatitis-a perspective.
Autoimmune hepatitis (AIH) is the inflammation of the liver with clear-cut interface hepatitis and piecemeal necrosis located at the boundary between portal areas and periportal hepatocytes, and characterized by autoimmunity to hepatocytes with an increase in the antinuclear antibody. After the disastrous SARS-CoV-2 pandemic flagellated several countries, several vaccines have been commercialized and have become a ground for social responsibility. The mRNA vaccines, issued by Pfizer-BioNTech (BNT162b2) and Moderna (mRNA-1273), do not use prebuilt viruses to supply the antigen in the subject's body and are not perfect but have been useful in tackling the pandemic. Nevertheless, both myocarditis and AIH have been reported as side effects of the vaccination programs in addition to thromboembolic events. Here, we explore this topic and give a data-based perspective, gathering a comparison between the titin protein of the sarcomere and myocarditis. The isolation of a Drosophila gene using the serum from a patient with autoimmune scleroderma recognized an epitope on chromosomes (condensed mitotic form) in both human cultured cells and early Drosophila embryos. It revealed that this gene encodes a Drosophila homolog of the vertebrate titin (D-Titin). Moreover, anti-titin antibodies have been found in a subset of patients with myasthenia gravis, a neuromuscular junction disease that is mostly associated with autoimmune antibodies, such as the anti-acetylcholine receptor antibody. The co-existence of myasthenia gravis and autoimmune hepatitis is rare, and a cohort of patients with myasthenia gravis anti-titin antibodies seems to be highly relevant. In consideration of these data and the number of patients who may not be symptomatic, we postulated that autoimmune phenomena may not be exceedingly rare, following the administration of mRNA technology-based vaccines, and a balance between pros and cons in administrating boosters is critical.


A cascade of care for people with epilepsy: learning from "HIV/AIDS 90-90-90".
Epilepsy is now more prevalent in many countries than HIV/AIDS. Building upon the advances of global policymaking for HIV/AIDS and creating a framework for countries and organizations to monitor progress in epilepsy care will help direct and justify much-needed novel programming. Given the clarity of the HIV/AIDS care continuum model and the UNAIDS 90-90-90 targets, I propose this same approach to the cascade of care could be used as a viable framework for people with epilepsy. In this model, the targets of success include (1) ensuring 90% of all people with epilepsy are aware of their diagnosis as a brain disorder, (2) starting 90% of people with epilepsy on quality controlled, appropriately chosen and well stocked antiepileptic drugs, and (3) achieving seizure freedom in 70% of those treated. At least 90% of all people with epilepsy must also be linked to and retained in appropriate care. Although the precise numbers may be debated, this cascade of care approach will assist in deconstructing the barriers to epilepsy care in populations better than the more familiar concept of the epilepsy treatment gap. These reflect concrete goals for health systems for epilepsy care that, if achieved, could lead to seizure freedom for the many people in lower income countries living with poorly controlled epilepsy.


Seizure-related hospital admissions, readmissions and costs: Comparisons with asthma and diabetes in South Australia.
Seizures are listed as an Ambulatory Care Sensitive Condition (ACSC), where, in some cases, hospitalisation may be avoided with appropriate preventative and early management in primary care. We examined the frequencies, trends and financial costs of first and subsequent seizure-related hospital admissions in the adult and paediatric populations, with comparisons to bronchitis/asthma and diabetes admissions in South Australia between 2012 and 2014. De-identified hospital separation data from five major public hospitals in metropolitan South Australia were analysed to determine the number of children and adults admitted for the following Australian Refined Diagnosis Related Groups: seizure related conditions; bronchitis/asthma; and diabetes. Additional data included length of hospital stay and type of admission. Demographic data were analysed to identify whether social determinants influence admission, and a macro costing approach was then applied to calculate the financial costs to the Health Care System. The rate of total seizure hospitalizations was 649 per 100,000; lower than bronchitis/asthma (751/100,000), yet higher than diabetes (500/100,000). The highest proportions of subsequent separations were recorded by children with seizures regardless of complexity (47% +CSCC; 17% -CSCC) compared with asthma (11% +CSCC; 14% -CSCC) or diabetes (14% +CSCC; 13% -CSCC), and by adults with seizures with catastrophic or severe complications/comorbidity (25%), compared with diabetes (22%) or asthma (14%). The mean cost per separation in both children and adults was highest for diabetes (AU$4438/$7656), followed by seizures (AU$2408/$5691) and asthma (AU$2084/$3295). Following the lead of well-developed and resourced health promotion initiatives in asthma and diabetes, appropriate primary care, community education and seizure management services (including seizure clinics) should be targeted in an effort to reduce seizure related hospitalisations which may be avoidable, minimise costs to the health budget, and maximise health care quality.


Antimicrobial resistance surveillance of Clostridioides difficile in Australia, 2015-18.
Clostridioides difficile was listed as an urgent antimicrobial resistance (AMR) threat in a report by the CDC in 2019. AMR drives the evolution of C. difficile and facilitates its emergence and spread. The C. difficile Antimicrobial Resistance Surveillance (CDARS) study is nationwide longitudinal surveillance of C. difficile infection (CDI) in Australia. To determine the antimicrobial susceptibility of C. difficile isolated in Australia between 2015 and 2018. A total of 1091 strains of C. difficile were collected over a 3 year period by a network of 10 diagnostic microbiology laboratories in five Australian states. These strains were tested for their susceptibility to nine antimicrobials using the CLSI agar incorporation method. All strains were susceptible to metronidazole, fidaxomicin, rifaximin and amoxicillin/clavulanate and low numbers of resistant strains were observed for meropenem (0.1%; 1/1091), moxifloxacin (3.5%; 38/1091) and vancomycin (5.7%; 62/1091). Resistance to clindamycin was common (85.2%; 929/1091), followed by resistance to ceftriaxone (18.8%; 205/1091). The in vitro activity of fidaxomicin [geometric mean MIC (GM) = 0.101 mg/L] was superior to that of vancomycin (1.700 mg/L) and metronidazole (0.229 mg/L). The prevalence of MDR C. difficile, as defined by resistance to ≥3 antimicrobial classes, was low (1.7%; 19/1091). The majority of C. difficile isolated in Australia did not show reduced susceptibility to antimicrobials recommended for treatment of CDI (vancomycin, metronidazole and fidaxomicin). Resistance to carbapenems and fluoroquinolones was low and MDR was uncommon; however, clindamycin resistance was frequent. One fluoroquinolone-resistant ribotype 027 strain was detected.


Longitudinal Associations of Neighborhood Crime and Perceived Safety With Blood Pressure: The Multi-Ethnic Study of Atherosclerosis (MESA).
High neighborhood crime and low perceptions of safety may influence blood pressure (BP) through chronic stress. Few studies have examined these associations using longitudinal data. We used longitudinal data from 528 participants of the Multi-Ethnic Study of Atherosclerosis (aged 45-84, nonhypertensive at baseline) who lived in Chicago, Illinois. We examined associations of changes in individual-level perceived safety, aggregated neighborhood-level perceived safety, and past-year rates of police-recorded crime in a 1, ½, or ¼ mile buffer per 1,000 population with changes in systolic and diastolic BPs using fixed-effects linear regression. BP was measured five times between 2000 and 2012 and was adjusted for antihypertensive medication use (+10 mm Hg added to systolic and +5 mm Hg added to diastolic BP for participants on medication). Models were adjusted for time-varying sociodemographic and health-related characteristics and neighborhood socioeconomic status. We assessed differences by sex. A standard deviation increase in individual-level perceived safety was associated with a 1.54 mm Hg reduction in systolic BP overall (95% confidence interval [CI]: 0.25, 2.83), and with a 1.24 mm Hg reduction in diastolic BP among women only (95% CI: 0.37, 2.12) in adjusted models. Increased neighborhood-level safety was not associated with BP change. An increase in police-recorded crime was associated with a reduction in systolic and diastolic BPs among women only, but results were sensitive to neighborhood buffer size. Results suggest individual perception of neighborhood safety may be particularly salient for systolic BP reduction relative to more objective neighborhood exposures.


Chiral Plasmonic Nanowaves by Tilted Assembly of Unidirectionally Aligned Block Copolymers with Buckling-Induced Microwrinkles.
Chiral-structured nanoscale materials exhibit chiroptical properties with preferential absorptions of circularly polarized light. The distinctive optical responses of chiral materials have great potential for advanced optical and biomedical applications. However, the fabrication of three-dimensional structures with mirrored nanoscale geometry is still challenging. This study introduces chiral plasmonic nanopatterns in wavy shapes based on the unidirectional alignment of block copolymer thin films and their tilted transfer, combined with buckling processes. The cylindrical nanodomains of polystyrene-block-poly(2-vinylpyridine) thin films were unidirectionally aligned over a large area by the shear-rolling process. The aligned block copolymer thin films were transferred onto uniaxially prestrained polydimethylsiloxane films at certain angles relative to the stretching directions. The strain was then released to induce buckling. The aligned nanopatterns across the axis of the formed microwrinkles were selectively infiltrated with gold ions. After reduction by plasma treatment, chiral plasmonic nanowave patterns were fabricated with the presence of mirror-reflected circular dichroism spectra. This fabrication method does not require any lithography processing or innately chiral biomaterials, which can be advantageous over other conventional fabrication methods for artificial nanoscale chiral materials.


Scanogram for sagittal imbalance of the spine: low dose alternative for a safer diagnosis.
The diagnosis of Fixed Sagittal Imbalance (FSI), previously known as Flat Back Syndrome, requires the measurement of spinal curvatures on a lateral radiograph in the standing position (C7-S1). It can be difficult to position a spastic patient, sometimes repeated exposure are required, at separate thoracic and lumbar levels, increasing the radiation dosage. CT Scanography is suggested as an alternative radiological diagnostic method since it is rapid to perform. The patient is comfortably positioned (horizontal) and it combines both prone and supine positions, therefore acting as a functional examination. This test was performed on 34 consecutive patients with fractured vertebrae (lumbar, dorsal) and with back pain persisting beyond the bone healing period. The functional scanogram was found to be accurate in diagnosing sagittal imbalances, but more importantly it offered reduction in radiation: in Entrance dose; in Effective dose and Absorption dose. Scanogram is therefore proposed as an alternative method for the diagnosis of FSI.


COD fractionation and denitrification potential of sonicated waste activated sludge liquids.
This study characterized sonicated waste activated sludge (WAS) liquids as a possible carbon source for nitrogen removal. In this context, the effect of sonication density on chemical oxygen demand (COD) and nitrogen release was determined by particle size distribution (PSD) analysis and anoxic batch experiments. The increase in ultrasonic density from 0.8 W/mL to 1.6 W/mL had a slight impact on the soluble COD/total COD ratio. The high ultrasonic energy input increased the solubilization of nitrogenous organic substances and resulted in a low COD/TKN (total Kjeldahl nitrogen) ratio, which is not appropriate for nutrient removal systems. The change in ultrasonic power had a significant effect on COD fractionation of sonicated WAS liquid. The COD fraction at the size ranges higher than 1600 nm decreased from 44% to 3% as the energy input increased. The increase in specific energy raised the COD fraction, at the size ranges of <2 nm, from 11% to 23%.The PSD-based COD fractionation showed that increasing the sonication density markedly changed the size distribution. The anoxic batch tests indicated that the specific denitrification rate of sonicated WAS liquid was in the range of that reported for the slowly biodegradable fraction of the domestic wastewater and higher than those reported for agro-food wastewater.


Evaluation of Functional Outcome Following Hybrid External Fixation in The Management of Schatzkers Type V and VI Tibial Plateau Fractures - A Prospective Study of 30 Patients.
Fractures of the proximal tibia are high velocity injuries often associated with soft tissue compromise especially in the type V and VI fracture patterns. Dual plating is the preferred treatment option for these injuries but not in a setting where there is extensive soft tissue injury, as this can lead to problems with wound healing. The aim of this study was to evaluate the functional outcome following hybrid external fixation in the management of Schatzkers type V and VI fractures. A total of 30 patients with type V and VI proximal tibial fractures who presented between January 2012 to January 2015 were managed with hybrid external fixation and were followed-up for a period of 3 years. The mean age of the patients was 42.26 years with the left knee being more commonly affected. Schatzkers type V was the more common fracture type seen. The mean time to union was 12.06 weeks and the average range of motion achieved was 0 to 100°. The mean Rasmussens functional score was 25.4 at last follow-up and we had excellent results in 5 patients and good results in 22 patients. Through this study, we conclude that the hybrid external fixation is an excellent option in the type V and VI fractures with extensive soft tissue compromise. It is easy to apply, facilitates early mobilisation of the joint and gives good functional results.


Characterization of brominated, chlorinated, and phosphate flame retardants in San Francisco Bay, an urban estuary.
Flame retardant chemical additives are incorporated into consumer goods to meet flammability standards, and many have been detected in environmental matrices. A uniquely wide-ranging characterization of flame retardants was conducted, including polybrominated diphenyl ethers (PBDEs) and 52 additional brominated, chlorinated, or phosphate analytes, in water, sediment, bivalves, and harbor seal blubber of San Francisco Bay, a highly urbanized estuary once considered a hot spot for PBDE contamination. Among brominated flame retardants, PBDEs remained the dominant contaminants in all matrices, though declines have been observed over the last decade following their phase-out. Hexabromocyclododecane (HBCD) and other hydrophobic, brominated flame retardants were commonly detected at lower levels than PBDEs in sediment and tissue matrices. Dechlorane Plus (DP) and related chlorinated compounds were also detected at lower levels or not at all across all matrices. In contrast, phosphate flame retardants were widely detected in Bay water samples, with highest median concentrations in the order TCPP > TPhP > TBEP > TDCPP > TCEP. Concentrations in Bay water were often higher than in other estuarine and marine environments. Phosphate flame retardants were also widely detected in sediment, in the order TEHP > TCrP > TPhP > TDCPP > TBEP. Several were present in bivalves, with levels of TDCPP comparable to PBDEs. Only four phosphate flame retardants were detected in harbor seal blubber: TCPP, TDCPP, TCEP, and TPhP. Periodic, multi-matrix screening is recommended to track contaminant trends impacted by changes to flammability standards and manufacturing practices, with a particular focus on contaminants like TDCPP and TPhP that were found at levels comparable to thresholds for aquatic toxicity.


Parent and Adolescent Thoughts About Suicide Risk Screening in Pediatric Outpatient Settings.
Suicide rates among adolescents in the United States continue to climb and many at-risk youths are undetected. Screening for suicidal thoughts has become the primary approach to identify those at risk, but no studies have assessed reactions to its deployment in pediatric outpatient settings. This mixed-method study assessed parents' and adolescents' thoughts about suicide risk screening in non-psychiatric, pediatric outpatient specialty settings.As part of a multi-site measurement validation study, adolescents (n = 269; ages 10-21) and parents (n = 246) at pediatric specialty clinics in the Midwest completed a survey regarding thoughts about suicide risk screening. Data were collected on tablet computers and transcribed verbatim. Three study team members independently coded transcripts of open-ended responses to identify major themes, and frequency data were analyzed using StataSE 15.1. Inter-rater agreement was substantial (Fleiss' Kappa ranged 75-86%).Parents (55% 41-50 years of age, 20% male, 80% White) and adolescents (Mean age = 14.3, 50% male, 77% White) agreed medical providers should screen adolescents for suicide risk (93% and 88%, respectively). Majority of parents indicated that the pediatric outpatient setting is appropriate for suicide risk screening. Major themes included the important role of providers in identifying at-risk youth, the potential for screening to prevent suicides, and concerns about iatrogenic risk and misdiagnosis.Most parents and adolescents support screening for suicide risk in pediatric outpatient settings. Nevertheless, some have concerns about the screening process and implications. As suicide risk screening becomes standard practice in adolescent care, it's critical to develop screening processes that maximize comfort and address concerns.


Pneumococcal disease seasonality: incidence, severity and the role of influenza activity.
We tested whether the effect of influenza activity on invasive pneumococcal disease incidence and severity varies between age and comorbidity groups. Weekly rates of invasive pneumococcal disease were obtained from the Danish National Laboratory Surveillance System (1977-2007). Influenza-like illness data were collected from a sentinel surveillance system at the Statens Serum Institut (Copenhagen, Denmark). We fitted Poisson regression models for invasive pneumococcal disease, with predictors of seasonality, trends and influenza activity, and allowed the influenza activity variable to vary by comorbidity level and clinical presentation. Influenza activity accounted for 8.4% (95% CI 4.8-11.9%) and 6.9% (95% CI 5.4-10.2%) of all invasive pneumococcal disease cases among those aged 15-39 and ≥40 years, respectively, but had no measurable impact among children aged <15 years. Influenza activity was associated with significant increases in the incidence of invasive pneumococcal pneumonia in both children and adults. The association was more pronounced among younger adults without comorbidities. Case fatality also varied seasonally among the elderly, and this variation might be associated with influenza activity. Pneumococcal incidence and the severity of disease varied seasonally and between age groups. The effect of influenza activity on pneumococcal disease varied between children and adults, and this difference was largely due to differences in disease presentation.


The anti-inflammatory effects of heparin and related compounds.
Heparin is a glycosaminoglycan well known for its anticoagulant properties. In addition, heparin possesses anti-inflammatory effects. Although the mechanisms responsible for the anticoagulant effects of heparin are well understood, those underlying its anti-inflammatory effects are not. This review presents some of the evidence from clinical and animal studies supporting an anti-inflammatory role for heparin and heparin-related derivatives. Potential mechanisms by which heparin can exert its anti-inflammatory effects are discussed. The clinical use of heparin as an anti-inflammatory agent has been held back by the fear of bleeding. Development of nonanticoagulant heparins or heparin derivatives should mitigate this concern.


Combined linkage and association mapping reveals CYCD5;1 as a quantitative trait gene for endoreduplication in Arabidopsis.
Endoreduplication is the process where a cell replicates its genome without mitosis and cytokinesis, often followed by cell differentiation. This alternative cell cycle results in various levels of endoploidy, reaching 4× or higher one haploid set of chromosomes. Endoreduplication is found in animals and is widespread in plants, where it plays a major role in cellular differentiation and plant development. Here, we show that variation in endoreduplication between Arabidopsis thaliana accessions Columbia-0 and Kashmir is controlled by two major quantitative trait loci, ENDO-1 and ENDO-2. A local candidate gene association analysis in a set of 87 accessions, combined with expression analysis, identified CYCD5;1 as the most likely candidate gene underlying ENDO-2, operating as a rate-determining factor of endoreduplication. In accordance, both the overexpression and silencing of CYCD5;1 were effective in changing DNA ploidy levels, confirming CYCD5;1 to be a previously undescribed quantitative trait gene underlying endoreduplication in Arabidopsis.


Comparative study of apoptosis induced by H(2)O(2) and NO: limitation of apoptosis induced by NO due to slower recovery of activity of NO-modified caspase.
Both H(2)O(2) and NO can act as apoptogens, triggering apoptosis in many cells. They are also well known inhibitors of caspases, essential enzymes in apoptosis. The differences between these two agents as apoptosis inducers and how caspases mediate apoptosis with these inhibitory agents is still unclear. Consistent with the previous reports, these two agents induced apoptosis accompanied by caspase activation with limitation of all apoptotic events for NO. It was found that NO-modified caspase-3 showed a slower recovery of its activity in the presence of the reducing agents compared to that of H(2)O(2) modification. This is one possible cause of the limited apoptosis in the case of NO.


The status of women in academic anesthesiology: a progress report.
The number of women in medicine has increased steadily in the last half century. In this study, we reassessed the status of women in academic anesthesiology departments in the United States in 2006. Medical student, resident, and faculty rank gender data were obtained from the Association of American Medical Colleges. Data regarding the make-up and gender of anesthesia subspecialty society leadership, the editorial boards of Anesthesia & Analgesia and Anesthesiology, the awardees of anesthesia research grants, American Board of Anesthesiology examiners, and department chairs were obtained from websites, organization management personnel, and the Wood Library-Museum of Anesthesiology. Anesthesiology data were compared with composite data from medical school departments in other clinical specialties and to data from previous years, beginning in 1985. The percentage of medical school graduates, anesthesiology residents, and anesthesiology faculty members who are women has increased since 1985; however, the rate of increase in the percentage of women is significantly faster for medical school graduates compared with anesthesiology residents (P < 0.001) and faculty (P < 0.05). The percentage of women anesthesiology faculty members who were full professors in 2006 was 6.5% compared with 17.7% of men faculty (P < 0.001) and is not significantly different than in 1986 (P = 0.27). Fourteen percent of full anesthesiology professors were women and this does not differ from all clinical specialties combined (15%). Women comprised 12.7% of academic anesthesiology chairs and 10% of all medical school department chairs in 2006, significantly higher compared with 1993 (P < 0.05). Currently, 8% and 11% of editors and associate editors of Anesthesiology and Anesthesia & Analgesia are women, respectively. Eighteen percent of American Board of Anesthesiology oral board examiners in 2007 were women compared with 8% in 1985 (P < 0.05). The percentage of time in which women have served as anesthesiology society leaders was significantly greater during 1997-2006 compared with 1987-1996 (P < 0.001). The proportion of competitive research grants awarded to women has not changed over several decades. The status of women in academic anesthesiology in the first decade of the millennium has, by some measures, advanced compared with 20 yr ago. However, by other measures, there has been no change. The task ahead is to identify factors that discourage qualified women medical students, residents, and junior faculty members from pursuing careers in academic anesthesiology and advancing in academic rank.


Molecular origin of time-dependent fluorescence shifts in proteins.
Time-resolved fluorescence spectroscopy is used increasingly to probe molecular motions at the aqueous interfaces of biological macromolecules and membranes. By recording the time variation of the fluorescence frequency, thermal atomic fluctuations in the vicinity of the chromophore can be probed. From such fluorescence Stokes shift (FSS) experiments, it has been inferred that water motions in the hydration layer are slowed down by 1-3 orders of magnitude. To provide a more secure foundation for the interpretation of FSS data, we use molecular dynamics simulations to examine the molecular origin of the FSS from a tryptophan residue in a protein. By using linear response theory to decompose the FSS into its water and protein components, we find that the water component dominates the static FSS but decays rapidly. Thus, after a few picoseconds, the FSS essentially reflects protein dynamics, including the self-motion of the chromophore. Because of its collective nature, the FSS response is insensitive to the motion of individual water molecules. Collective water displacement by slowly fluctuating protein groups introduces a long-time tail in the water autocorrelation function, but this dynamic coupling is hardly manifested in the observed FSS. Our analysis reconciles FSS data with the picture of a highly dynamic hydration layer, derived mainly from magnetic relaxation dispersion and simulation studies, and calls for a revision of previous interpretations of FSS decays in terms of slow hydration dynamics at biomolecular and other interfaces.


Impact of drug price adjustments on utilization of and expenditures on angiotensin-converting enzyme inhibitors and angiotensin receptor blockers in Taiwan.
A previous study has suggested that drug price adjustments allow physicians in Taiwan to gain greater profit by prescribing generic drugs. To better understand the effect of price adjustments on physician choice, this study used renin-angiotensin drugs (including angiotensin-converting enzyme inhibitors [ACEIs] and angiotensin receptor blockers [ARBs]) to examine the impact of price adjustments on utilization of and expenditures on patented and off-patent drugs with the same therapeutic indication. Using the Taiwan's Longitudinal Health Insurance Database (2005), we identified 147,157 patients received ACEIs and/or ARBs between 1997 and 2008. The annual incident and prevalent users of ACEIs, ARBs and overall renin-angiotensin drugs were examined. Box-Tiao intervention analysis was applied to assess the impact of price adjustments on monthly utilization of and expenditures on these drugs. ACEIs were divided into patented and off-patent drugs, off-patent ACEIs were further divided into original brands and generics, and subgroup analyses were performed. The number of incident renin-angiotensin drug users decreased over the study period. The number of prevalent ARB users increased and exceeded the cumulative number of first-time renin-angiotensin drug users starting on ARBs, implying that some patients switched from ACEIs to ARBs. After price adjustments, long term trend increases in utilization were observed for patented ACEIs and ARBs; a long-term trend decrease was observed for off-patent ACEIs; long-term trend change was not significant for overall renin-angiotensin drugs. Significant long-term trend increases in expenditures were observed for patented ACEIs after price adjustment in 2007 (200.9%, p = 0.0088) and in ARBs after price adjustments in 2001 (173.4%, p < 0.0001) and 2007 (146.3%, p < 0.0001). A significant long-term trend decrease in expenditures was observed for off-patent ACEIs after 2004 price adjustment (-156.9%, p < 0.0001). Expenditures on overall renin-angiotensin drugs showed long-term trend increases after price adjustments in 2001 (72.2%, p < 0.0001) and 2007 (133.4%, p < 0.0001). Price adjustments did not achieve long-term cost savings for overall renin-angiotensin drugs. Possible switching from ACEIs to ARBs within individuals is evident. Policy makers should reconsider the appropriateness of the current adjustment strategies applied to patented and off-patent drugs.


Role of imaging in cardiac stem cell therapy.
Stem cell therapy has emerged as a potential therapeutic option for cell death-related heart diseases. Preclinical and a number of early phase human studies suggested that cell therapy may augment perfusion and increase myocardial contractility. The rapid translation into clinical trials has left many issues unresolved, and emphasizes the need for specific techniques to visualize the mechanisms involved. Furthermore, the clinical efficacy of cell therapy remains to be proven. Imaging allows for in vivo tracking of cells and can provide a better understanding in the evaluation of the functional effects of cell-based therapies. In this review, a summary of the most promising imaging techniques for cell tracking is provided. Among these are direct labeling of cells with super-paramagnetic agents, radionuclides, and the use of reporter genes for imaging of transplanted cells. In addition, a comprehensive summary is provided of the currently available studies investigating a cell therapy-related effect on left ventricular function, myocardial perfusion, scar tissue, and myocardial viability.


An Update on Protein Kinases as Therapeutic Targets-Part II: Peptides as Allosteric Protein Kinase C Modulators Targeting Protein-Protein Interactions.
Human protein kinases are highly-sought-after drug targets, historically harnessed for treating cancer, cardiovascular disease, and an increasing number of autoimmune and inflammatory conditions. Most current treatments involve small molecule protein kinase inhibitors that interact orthosterically with the protein kinase ATP-binding pocket. As a result, these compounds are often poorly selective and highly toxic. Part I of this series reviews the role of PKC isoforms in various human diseases, featuring cancer and cardiovascular disease, as well as translational examples of PKC modulation applied to human health and disease. In the present Part II, we discuss alternative allosteric binding mechanisms for targeting PKC, as well as novel drug platforms, such as modified peptides. A major goal is to design protein kinase modulators with enhanced selectivity and improved pharmacological properties. To this end, we use molecular docking analysis to predict the mechanisms of action for inhibitor-kinase interactions that can facilitate the development of next-generation PKC modulators.


Five-year outcomes in kidney transplant patients converted from cyclosporine to everolimus: the randomized ZEUS study.
ZEUS study was an open-label, 12-month, multicenter study in which 300 de novo kidney transplant recipients were randomized to continue receiving cyclosporine (CsA) or convert to everolimus at 4.5 months posttransplant. Five-year follow-up data were available for 245/269 patients (91.1%) who completed the core 12-month study (123 everolimus, 109 CsA). At 5 years, adjusted estimated GFR was 66.2 mL/min/1.73 m(2) with everolimus versus 60.9 mL/min/1.73 m(2) with CsA; the mean difference was 5.3 mL/min/1.73 m(2) in favor of everolimus (95% CI 2.4, 8.3; p < 0.001 [intent-to-treat population]). In a post hoc analysis of patients remaining on study drug at 5 years (everolimus 77, CsA 86), mean difference was 8.2 mL/min/1.73 m(2) (95% CI 4.3, 12.1; p < 0.001) in favor of everolimus. The cumulative incidence of biopsy-proven acute rejection postrandomization was 13.6% with everolimus versus 7.5% with CsA (p = 0.095), largely accounted for by grade I rejection (16/21 patients and 7/11 patients, respectively). Postrandomization, graft loss, mortality, serious adverse events and neoplasms were similar in both arms. In conclusion, conversion of kidney transplant patients to everolimus at 4.5 months posttransplant is associated with a significant improvement in renal function that is maintained to at least 5 years. The increase in early mild acute rejection did not affect long-term graft function.


[Breast cancer mortality trend in Portugal (1955-2002)].
In the last decade there were significant changes in breast cancer mortality that is declining since the 90 s in several countries. The aim of this study was to quantify the variation of the female breast cancer mortality rates, in Portugal and in different regions of the country, considering the changes in the trends. Breast cancer mortality rates and the number of deaths were obtained from the World Health Organization, for the years from 1955 to 2002, and the number of deaths and population in each region were obtained from the publication Risco de Morrer em Portugal. Age-standardized rates were computed by the direct method, using the European standard population. Joinpoint regression analyses were performed to identify the years in which changes in trends have occurred, and to estimate the annual percent changes in each period, for the age groups 35-74, 35-44, 45-54, 55-64 and 65-74 years. The annual variation was computed for each Portuguese region from 1990 to 2002. Breast cancer mortality rates increased 1.55%/year (95% Confidence Interval (CI): 1.43 to 1.68) between 1955 and 1992 and changed -2.02%/year (95%CI: -2.81 to -1.31) from 1992 to 2002 in women aged 35 to 74 years. No significant differences were observed in the joinpoints or the estimated annual percent changes across age strata. At a regional level, the mortality trends between 1990 and 2002 were inversely associated with the rates at the beginning of this period (r= -0.74, p <0.001), the larger decline being observed in Lisbon, with rates varying -3.58%/ year (95% CI: -5.30 to -1.48). A decrease in breast cancer mortality has been observed in Portugal in the early 90 s, with an estimated annual percent change of -2%/year in the last decade. The decline in mortality correlated positively with the rates at the beginning of the period.


Nephrology training programs and applicants: a very good match.
A critical mission of academic nephrology programs is training the next generation of nephrologists. Attracting high-quality candidates into nephrology training programs is the first step in this process. The training program directors (TPDs) in nephrology launched an initiative several years ago to standardize the application and recruitment process for applicants to their programs. This has recently culminated in the completion of a formal matching process using the Medical Specialties Matching Program (MSMP). This review will outline the history of this process and present the results of the recent matching process in nephrology for applicants who applied to training programs with a start date of July 1, 2009. The authors report that 89% of participating programs and 95% of applicant positions were filled. On the basis of a survey of nephrology TPDs, the match process was successful, with almost all respondents planning on participating in the match with a start date of July 1, 2010. Certain issues remain, including determining the correct number of interviews, how many applicants to rank, and whether four tracks are necessary. It is concluded that the nephrology match is of benefit to both training programs and applicants.


The National Acoustic Laboratories' procedure for selecting the saturation sound pressure level of hearing aids: theoretical derivation.
This paper presents the derivation of a procedure for prescribing the saturation sound pressure level (SSPL) of hearing aids. The procedure is designed to be used with either measured values of loudness discomfort level (LDL) or with hearing threshold values alone. SSPL needs to be low enough to prevent the hearing aid from causing loudness discomfort to the aid wearer but high enough to prevent the hearing aid from being excessively saturated by speech. The maximum SSPL likely to be acceptable can be predicted by measuring LDL or by estimating LDL from hearing thresholds. The minimum SSPL likely to be acceptable can be predicted by calculating, for any particular hearing loss, the amount of gain likely to be needed and hence the SSPL needed if the speech input signal is continuous discourse at an overall level of 75 dB SPL. The midpoint between the minimum and maximum acceptable SSPL values is defined as the optimal or prescribed SSPL, and the three frequency average (3FA; 500, 1000, and 2000 Hz) value of this can be predicted from the 3FA hearing thresholds. Alternatively, the SSPL prescription at each frequency can be prescribed on the basis of the hearing aid gain at each frequency. For either method, the SSPL prescription needs to be increased for people with a conductive component to their hearing losses. The SSPL prescription, when referred to a 2-cc coupler, needs to be decreased for infants, for deeply inserted hearing aids, for multichannel hearing aids that limit SSPL separately in each band, and possibly for nonlinear hearing aids. The 3FA SSPL prescribed for persons with a sensorineural hearing loss increases linearly from 89 dB SPL for normal hearing to 107 dB SPL for a person with a 60 dB HL 3FA loss, and then linearly again to 139 dB SPL for a person with a 120 dB HL 3FA loss. The procedure predicts that the acceptable range of SSPLs is very wide for people with mild losses but is vanishingly small for people with profound losses.


Effects of a multi-strain probiotic supplement for 12 weeks in circulating endotoxin levels and cardiometabolic profiles of medication naïve T2DM patients: a randomized clinical trial.
The present randomized clinical trial characterized the beneficial effects of a multi-strain probiotics supplementation on improving circulating endotoxin levels (primary endpoint) and other cardiometabolic biomarkers (secondary endpoint) in patients with T2DM. A total of 78 adult Saudi T2DM patients (naïve and without co-morbidities) participated in this clinical trial and were randomized to receive twice daily placebo or probiotics [(2.5 × 109 cfu/g) containing the following bacterial strains: Bifidobacterium bifidum W23, Bifidobacterium lactis W52, Lactobacillus acidophilus W37, Lactobacillus brevis W63, Lactobacillus casei W56, Lactobacillus salivarius W24, Lactococcus lactis W19 and Lactococcus lactis W58 (Ecologic®Barrier)] in a double-blind manner for 12 weeks. Anthropometrics and cardiometabolic profiles were obtained at baseline and after 12/13 weeks of treatment. After 12/13 weeks of intervention and using intention-to-treat analysis, no difference was noted in endotoxin levels between groups [Placebo - 9.5% vs. Probiotics - 52.2%; (CI - 0.05 to 0.36; p = 0.15)]. Compared with the placebo group however, participants in the probiotics groups had a significant but modest improvement in WHR [Placebo 0.0% vs. Probiotics 1.11%; (CI - 0.12 to - 0.01; p = 0.02)] as well as a clinically significant improvement in HOMA-IR [Placebo - 12.2% vs. Probiotics - 60.4%; (CI - 0.34 to - 0.01; p = 0.04)]. Using a multi-strain probiotic supplement daily for 12/13 weeks significantly improved HOMA-IR and modestly reduced abdominal adiposity among medication naïve T2DM patients. ClinicalTrials.gov Identifier: NCT01765517 , Registered January 10, 2013.


[Studies on the glucocorticoid receptor in human peripheral lymphocytes. III. Correlation between receptor number and clinical response to glucocorticoid].
Although the clinical response to glucocorticoid (GC) therapy varied considerably from patient to patient, it is not known how to predict the responsiveness of each patient. In the present study, it was evaluated whether the responsiveness to GC in an individual patient was predictable by the GC receptor number. GC receptor number in human peripheral lymphocytes was measured by whole cell binding assay using tritiated dexamethasone as a ligand. There was a significant negative correlation between receptor number and dose of GC being given to the patient (down-regulation). The relative receptor number was expressed as a percentage of the receptor number to the mean of the receptor number in the group of patients stratified by every 10mg/day of the dosage of prednisolone. This relative receptor number was constant in an individual subject regardless of GC dosage. In 12 patients with lupus nephritis who had a high receptor number (more than 100%), 9 patients (83%) showed improvement in their urinary findings after GC treatment. On the other hand, only 3 of 11 lupus nephritis patients with a low receptor number (less than 100%) showed improvement after GC therapy. In patients with various collagen diseases treated with 40-60 mg/day of prednisolone, there were significant correlations between receptor number and each of % creatinuria, total cholesterol in serum, and fasting urinary calcium excretion, which were known to be affected by GC administration. Thus, the relative receptor number in lymphocytes was well correlated with both therapeutic and metabolic effects of GC in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)


Salt-induced stabilization of apoflavodoxin at neutral pH is mediated through cation-specific effects.
Electrostatic contributions to the conformational stability of apoflavodoxin were studied by measurement of the proton and salt-linked stability of this highly acidic protein with urea and temperature denaturation. Structure-based calculations of electrostatic Gibbs free energy were performed in parallel over a range of pH values and salt concentrations with an empirical continuum method. The stability of apoflavodoxin was higher near the isoelectric point (pH 4) than at neutral pH. This behavior was captured quantitatively by the structure-based calculations. In addition, the calculations showed that increasing salt concentration in the range of 0 to 500 mM stabilized the protein, which was confirmed experimentally. The effects of salts on stability were strongly dependent on cationic species: K(+), Na(+), Ca(2+), and Mg(2+) exerted similar effects, much different from the effect measured in the presence of the bulky choline cation. Thus cations bind weakly to the negatively charged surface of apoflavodoxin. The similar magnitude of the effects exerted by different cations indicates that their hydration shells are not disrupted significantly by interactions with the protein. Site-directed mutagenesis of selected residues and the analysis of truncation variants indicate that cation binding is not site-specific and that the cation-binding regions are located in the central region of the protein sequence. Three-state analysis of the thermal denaturation indicates that the equilibrium intermediate populated during thermal unfolding is competent to bind cations. The unusual increase in the stability of apoflavodoxin at neutral pH affected by salts is likely to be a common property among highly acidic proteins.


Functional analysis of Hes-1 in preadipocytes.
Notch signaling blocks differentiation of 3T3-L1 preadipocytes, and this can be mimicked by constitutive expression of the Notch target gene Hes-1. Although considered initially to function only as a repressor, recent evidence indicates that Hes-1 can also activate transcription. We show here that the domains of Hes-1 needed to block adipogenesis coincide with those necessary for transcriptional repression. HRT1, another basic-helix-loop-helix protein and potential Hes-1 partner, was also induced by Notch in 3T3-L1 cells but did not block adipogenesis, suggesting that Hes-1 functions primarily as a homodimer or possibly as a heterodimer with an unknown partner. Purification of Hes-1 identified the Groucho/transducin-like enhancer of split family of corepressors as the only significant Hes-1 interacting proteins in vivo. An evaluation of global gene expression in preadipocytes identified approximately 200 Hes-1-responsive genes comprising roughly equal numbers of up-regulated and down-regulated genes. However, promoter analyses indicated that the down-regulated genes were significantly more likely to contain Hes-1 binding sites, indicating that Hes-1 is more likely to repress transcription of its direct targets. We conclude that Notch most likely blocks adipogenesis through the induction of Hes-1 homodimers, which repress transcription of key target genes.


Effects of body size and gender on the population pharmacokinetics of artesunate and its active metabolite dihydroartemisinin in pediatric malaria patients.
Despite the important role of the antimalarial artesunate and its active metabolite dihydroartemisinin (DHA) in malaria treatment efforts, there are limited data on the pharmacokinetics of these agents in pediatric patients. This study evaluated the effects of body size and gender on the pharmacokinetics of artesunate-DHA using data from pediatric and adult malaria patients. Nonlinear mixed-effects modeling was used to obtain a base model consisting of first-order artesunate absorption and one-compartment models for artesunate and for DHA. Various methods of incorporating effects of body size descriptors on clearance and volume parameters were tested. An allometric scaling model for weight and a linear body surface area (BSA) model were deemed optimal. The apparent clearance and volume of distribution of DHA obtained with the allometric scaling model, normalized to a 38-kg patient, were 63.5 liters/h and 65.1 liters, respectively. Estimates for the linear BSA model were similar. The 95% confidence intervals for the estimated gender effects on clearance and volume parameters for artesunate fell outside the predefined no-relevant-clinical-effect interval of 0.75 to 1.25. However, the effect of gender on apparent DHA clearance was almost entirely contained within this interval, suggesting a lack of an influence of gender on this parameter. Overall, the pharmacokinetics of artesunate and DHA following oral artesunate administration can be described for pediatric patients using either an allometric scaling or linear BSA model. Both models predict that, for a given artesunate dose in mg/kg of body weight, younger children are expected to have lower DHA exposure than older children or adults.


Hemodynamic support in the early phase of septic shock: a review of challenges and unanswered questions.
Improving sepsis support is one of the three pillars of a 2017 resolution according to the World Health Organization (WHO). Septic shock is indeed a burden issue in the intensive care units. Hemodynamic stabilization is a cornerstone element in the bundle of supportive treatments recommended in the Surviving Sepsis Campaign (SSC) consecutive biannual reports. The "Pandera's box" of septic shock hemodynamics is an eternal debate, however, with permanent contentious issues. Fluid resuscitation is a prerequisite intervention for sepsis rescue, but selection, modalities, dosage as well as duration are subject to discussion while too much fluid is associated with worsen outcome, vasopressors often need to be early introduced in addition, and catecholamines have long been recommended first in the management of septic shock. However, not all patients respond positively and controversy surrounding the efficacy-to-safety profile of catecholamines has come out. Preservation of the macrocirculation through a "best" mean arterial pressure target is the actual priority but is still contentious. Microcirculation recruitment is a novel goal to be achieved but is claiming more knowledge and monitoring standardization. Protection of the cardio-renal axis, which is prevalently injured during septic shock, is also an unavoidable objective. Several promising alternative or additive drug supporting avenues are emerging, trending toward catecholamine's sparing or even "decatecholaminization." Topics to be specifically addressed in this review are: (1) mean arterial pressure targeting, (2) fluid resuscitation, and (3) hemodynamic drug support. Improving assessment and means for rescuing hemodynamics in early septic shock is still a work in progress. Indeed, the bigger the unresolved questions, the lower the quality of evidence.


Association of polycystic ovary syndrome and a non-dipping blood pressure pattern in young women.
The association between polycystic ovarian syndrome and increased cardiovascular disease risk is still a controversial issue. In light of data documenting some common pathways or common end-points, the present study was undertaken to determine whether there is a relationship between sleep blood pressure pattern disturbances and polycystic ovarian syndrome in young women. The daytime and nighttime ambulatory blood pressures (BPs) were determined for each subject, according to the actual waking and sleeping times recorded in their individual diaries, in this cross-sectional study. The study group comprised 168 women (mean age: 25.7+/-5.5) diagnosed with polycystic ovarian syndrome, while the control group included 52 age- and BMI-matched healthy subjects (mean age: 26.1+/-5.4). When nocturnal BP declines very little or not at all, with the BP falling less than 10% during sleep compared with waking values, this pattern is classified as a non-dipping BP pattern. However, the non-dipping pattern of BP changes was significantly more common in polycystic ovarian syndrome patients compared to the control group (p<0.01). The prevalence of a non-dipping BP pattern was 43.4% (73 patients) in polycystic ovarian syndrome patients and 3.9% (2 patients) in the control group. Our cross-sectional study revealed that a non-dipping BP pattern is highly prevalent in polycystic ovarian syndrome patients, even if they are young and non-obese.


Leishmania and the Model of Predominant Clonal Evolution.
As it is the case for other pathogenic microorganisms, the respective impact of clonality and genetic exchange on Leishmania natural populations has been the object of lively debates since the early 1980s. The predominant clonal evolution (PCE) model states that genetic exchange in these parasites' natural populations may have a high relevance on an evolutionary scale, but is not sufficient to erase a persistent phylogenetic signal and the existence of bifurcating trees. Recent data based on high-resolution markers and genomic polymorphisms fully confirm the PCE model down to a microevolutionary level.


B lymphocytes, the gastrointestinal tract and autoimmunity.
Under homeostatic conditions, bidirectional interactions between the gastrointestinal and the immune system allow production of both inflammatory and anti-inflammatory responses designed to prevent undesirable inflammation and to respond efficiently to potential insults. This balanced regulation can be disrupted in disorders that affect tissues remote to the gastrointestinal tract, as seen in autoimmune diseases. Recent reports have described a variety of B lymphocyte-mediated functions that likely contribute to gastrointestinal homeostasis to a greater extent than previously thought. Studies have shown that early B cell development takes place within the intestine, and that self-reactive B cells are rendered tolerant using mechanisms known to occur in the bone marrow, indicating that the gastrointestinal tract contributes to maintaining immune tolerance to self. Relatedly, continuous bacterial stimulation is essential for maintaining regulatory B cell functions and for mediating mucosal homeostasis. In studies of neuro-inflammation, intestinal IgA+ B cells, which constitute a prominent source of lymphocytes in the organism, can migrate to inflamed tissues and exert regulatory functions that attenuate inflammation in the central nervous system, indicating that, in addition to its local effects in the intestin, gut microbiota-B cell crosstalk can exert long-range beneficial effects. At the translational level, metabolites produced by gut microbiota can act as B cell-intrinsic epigenetic modulators, reducing inflammation in the skin and kidneys of mice suffering from experimental lupus. Given the significant impact of B cell-intestinal microbiota interactions, there is a momentum for improving our understanding of these pathways in autoinflammatory diseases and for designing novel therapeutic strategies for systemic autoimmune diseases where B cells play key roles.


Wernicke's encephalopathy manifested as Korsakoff's syndrome in a patient with promyelocytic leukemia.
A case of Wernicke's encephalopathy associated with promyelocytic leukemia found at autopsy is reported. The patient was 30 years old and was undergoing chemotherapeutic treatment when she had a memory deficit for recent events (Korsakoff's syndrome) which persisted for 6 months, until death. The neuropathologic examination showed typical, old lesions that characterize Wernicke's encephalopathy, but only in the mamillary bodies. This case is compared with three other cases of Wernicke's encephalopathy associated with leukemia previously described in the literature. Comment is made on the cliniconeuropathologic picture and risk factors in leukemic patients that may favor the appearance of Wernicke's encephalopathy.


Negative role of surface plasmons in the transmission of metallic gratings with very narrow slits.
It is generally admitted that the extraordinary transmission of metallic grating with very narrow slits is mainly due to the excitation of surface plasmons on the upper and lower interfaces of the grating. We show that the surface plasmon contribution is not the prime effect and that waveguide mode resonance and diffraction are responsible for the extraordinary transmission. Additionally and surprisingly, we reveal that the transmittance of subwavelength metallic gratings is always nearly zero for frequencies corresponding to surface plasmon excitation. This finding implies that surface plasmons play a negative role in the transmission.


Learning how we learn: an ethnographic study in a neonatal intensive care unit.
This paper is a report of a study to identify how nurse clinicians learn with and from each other in the workplace. Clinicians' everyday practices and interactions with each other have recently been targeted as areas of research, because it is there that quality of care and patient safety are achieved. Orientation of new nurses and doctors into a specialty unit often results in stress. An ethnographic approach was used, including a 12-month period of fieldwork observations involving participation and in-depth interviews with nurse, doctor and allied health clinicians in their workplace. The data were collected in 2005-2006 in a paediatric teaching hospital in Australia. The findings were grouped into four dimensions: orientation of nurses, orientation of medical registrars, preceptoring and decision-making. The orientation of new staff (nursing and medical) is a complex and multi-layered process which accommodates multiple kinds of learning, in addition to formal learning. Workplace learning also can be informal, incidental, interpersonal and interactive. Interactive and interpersonal learning and the transfer of knowledge include codified and tacit knowledge as well as intuitive understandings of 'how we do things here'. Research into how nurses learn is crucial for illuminating learning that is non-formal and less recognized than more formal kinds. To provide a safe practice environment built on a foundation of knowledge and best practice, there needs to be an allocation of time in the busy workday for learning and reflection.


Cytokine Adsorption Therapy during Extracorporeal Membrane Oxygenation in Adult Patients with COVID-19.
Respiratory failure and systemic inflammation are paramount features of severe SARS-CoV-2 disease (COVID-19). Extracorporeal membrane oxygenation (ECMO) therapy has a potential role in patients with refractory disease. An inflammatory response due to blood contact with hemofilters, functioning as a synergic inflammatory stimulus, can lead to a hyperinflammatory state, relatable to cytokine release syndromes. After the first patient succumbed to a refractory vasodilatory shock believed to be due to hyperinflammatory state, a strategy of blood purification through cytokine adsorption therapy (CAT) with CytoSorb® was designed. In this case series, the authors describe the initial experience with such strategy. CAT was employed with no direct complications and helped controlling the inflammatory state, with all patients halting vasopressor support in 72 h and biomarker levels (C-reactive protein, ferritin, and interleukin-6) showing negative trends in most patients. Analysis of inflammatory biomarkers evolution highlighted 2 biomarker profiles related to the presence or absence of superinfection at the time of CAT implementation. In this case series of severe COVID-19 patients, 3 patients died - irreversible lung fibrosis, complications of critical hypoxemia before ECMO induction and complications of systemic anticoagulation were the causes. This case series aimed to contribute to the body of evidence substantiating CAT utilization in hyperinflammatory patients, namely, COVID-19 patients requiring ECMO rescue.


Male reproductive senescence: the price of immune-induced oxidative damage on sexual attractiveness in the blue-footed booby.
In animals, male reproduction is commonly a function of sexual attractiveness, based on the expression of sexually dimorphic traits that advertise genuinely the male's quality. Male performance may decline with age because physiological functions underlying sexual attractiveness may be affected by senescence. Here we show that a sexual signal (foot colour) declines with age, due probably to the deleterious effects of oxidative damage. We found that in the blue-footed booby Sula nebouxii foot colour during courtship was less attractive in senescent than in middle-aged males. In addition, we increased reactive oxygen species experimentally by immunizing males with lipopolysaccharide, a bacterial cell wall component that induces marked oxidative stress in animals. The immune system activation induced greater lipid peroxidation and invoked changes on colour expression (less attractive), particularly in senescent males. These results support the idea that oxidative stress affects reproductive senescence, and suggest that oxidative damage might be a proximal mechanism underlying age-reproductive patterns in long-lived animals.


Attempt to develop live attenuated bacterial vaccines by selecting resistance to gossypol, proflavine hemisulfate, novobiocin, or ciprofloxacin.
In an attempt to develop attenuated bacteria as potential live vaccines, four chemicals (gossypol, proflavine hemisulfate, novobiocin, and ciprofloxacin) were used to modify the following four genera of bacteria through chemical-resistance strategy: (1) Aeromonas hydrophila (9 isolates); (2) Edwardsiella tarda (9 isolates); (3) Streptococcus iniae (9 isolates); and (4) S. agalactiae (11 isolates). All bacteria used in this study were able to develop high resistance to gossypol. However, only some bacteria were able to develop resistance to proflavine hemisulfate, novobiocin, or ciprofloxacin. When the virulence of resistant bacteria was tested in tilapia or catfish, none of the gossypol-resistant isolate was attenuated, whereas majority of the proflavine hemisulfate-resistant isolates were attenuated. However, all proflavine hemisulfate-attenuated bacteria failed to provide significant protection to fish. Eight novobiocin- or ciprofloxacin-resistant Gram-positive bacteria (S. agalactiae and S. inaie) were found to be attenuated. However, none of them offered protection higher than 70%. Of seven attenuated novobiocin- or ciprofloxacin-resistant Gram-negative isolates (A. hydrophila and E. tarda), only one (novobiocin-resistant E. tarda 30305) was found to safe and highly efficacious. When E. tarda 30305-novo vaccinated Nile tilapia were challenged by its virulent E. tarda 30305, relative percent of survival of vaccinated fish at 14- and 28-days post vaccination (dpv) was 100% and 92%, respectively. Similarly, E. tarda 30305-novo offered 100% protection to channel catfish against challenges with virulent parent isolate E. tarda 30305 at both 14- and 28-dpv. Our results suggest that the development of live attenuated bacterial vaccines that are safe and efficacious is challenging, although it is feasible.


IgA responses in xid mice: oral antigen primes Peyer's patch cells for in vitro immune responses and secretory antibody production.
Because the gut-associated lymphoreticular tissue (GALT), e.g., Peyer's patches (PP), of X-linked immunodeficient (xid) mice possesses a subpopulation of mature B cells, we have characterized the ability of xid mice to respond to the thymic-dependent antigen sheep erythrocytes (SRBC) given by the oral route. Gastric intubation of SRBC to xid (CBA/N X DBA/2) F1 male or CBA/N mice, followed by the in vitro culture of dissociated PP cells with SRBC, resulted in IgM, IgG1, IgG2, and high IgA anti-SRBC plaque-forming cell (PFC) responses. The addition of unprimed PP but not splenic T cells to splenic xid B cell cultures resulted in IgM anti-SRBC PFC responses, suggesting the importance of GALT T cells for support of the immune responses to SRBC by splenic B cells from xid mice. Furthermore, purified PP T cells from SRBC orally primed xid mice supported in vitro IgA anti-SRBC PFC responses in B cell cultures from either the PP or the spleens of nonprimed xid mice. Higher IgA responses, however, occurred in PP, when compared with splenic B cell cultures. Additional evidence that the GALT of xid mice contains functional IgA precursor cells was provided by the finding that cloned H-2k PP T helper cells (PP Th A) supported IgA responses in PP B cell cultures derived from (CBA/N X C3H/HeN) F1 male (xid) mice. On the other hand, splenic B cells from these xid mice, in the presence of PP Th A cells, did not support in vitro responses. These results suggest that unique subpopulations of T cells occur in the GALT of xid and normal mice; one T cell subpopulation may induce immature B cells to become precursor IgA cells in the PP. A separate GALT T cell subpopulation, e.g., isotype-specific helper T cells, effectively collaborates with mature IgA B cells for the induction of IgA responses to orally administered antigen. When xid mice were gastric intubated with SRBC, followed by i.p. injection of SRBC, good splenic IgA anti-SRBC PFC responses were seen. Salivary and serum IgA antibodies were also detected in these xid mice. Nevertheless, the magnitude of the anti-SRBC response in xid mice was lower than that seen in similarly treated normal mice. These studies indicate that the GALT of both xid and normal mice possess unique populations of T cells that support in vitro responses in xid B cell cultures from either the spleen or the PP, which direct the mature B cell populations present toward IgA isotype-specific responses.(ABSTRACT TRUNCATED AT 400 WORDS)


Triggering Mechanisms for Motor Actions: The Effects of Expectation on Reaction Times to Intense Acoustic Stimuli.
Motor actions can be released much sooner than normal when the go-signal is of very high intensity (>100 dBa). Although statistical evidence from individual studies has been mixed, it has been assumed that sternocleidomastoid (SCM) muscle activity could be used to distinguish between two neural circuits involved in movement triggering. We summarized meta-analytically the available evidence for this hypothesis, comparing the difference in premotor reaction time (RT) of actions where SCM activity was elicited (SCM+ trials) by loud acoustic stimuli against trials in which it was absent (SCM- trials). We found ten studies, all reporting comparisons between SCM+ and SCM- trials. Our mini meta-analysis showed that premotor RTs are faster in SCM+ than in SCM- trials, but the effect can be confounded by the variability of the foreperiods employed. We present experimental data showing that foreperiod predictability can induce differences in RT that would be of similar size to those attributed to the activation of different neurophysiological pathways to trigger prepared actions. We discuss plausible physiological mechanisms that would explain differences in premotor RTs between SCM+ and SCM- trials.


Complete atrioventricular block after self-ingestion of Nerium oleander for relief of hemorrhoidal complaints.
Nerium oleander is a plant native only in the Mediterranean region, but it can also be cultivated worldwide, particularly in warm areas. Biologically active oleander compounds may be used for therapeutic purposes. However, when used for self-medication, it may cause serious problems including death. We present a 30-year-old otherwise healthy man who developed complete atrioventricular block after taking a syrup of N. oleander leaves for self-medication to relive hemorrhoidal complaints. The patient was treated by oral administration of charcoal combined with sodium sulfate as well as electrolyte solutions and transient use of an external cardiac pacemaker. The atrioventricular block reverted to sinus rhythm in 30 hours and he was discharged in good hemodynamic status and general condition.


The risk of foot ulceration in people with diabetes screened in community settings: findings from a cohort study.
Annual foot checks are recommended in patients with diabetes mellitus (DM) to identify those at risk of foot ulceration. Systematic reviews have found few studies evaluating the predictive value of tests in community-based diabetic populations. To quantify the predictive value of clinical risk factors in relation to foot ulceration in a community population. A cohort of 1192 people with diabetes receiving care in community settings was recruited and a screening procedure, covering symptoms, signs and diagnostic tests was conducted at baseline. At an average 1-year follow-up patients who developed a foot ulcer were identified by an independent blind assessor. Multivariable analysis was performed to identify clinical predictors of foot ulceration. The incidence of foot ulceration was 1.93% [95% confidence interval (CI) 1.27-2.89). Three time-independent clinical predictors with five factors were selected: previous amputation [odds ratio (OR) 14.7, 95% CI 3.1-69.5), use of insulin before 3 months with inability to distinguish between cool and cold temperatures (OR 2.97, 95% CI 1.9-4.5) and failure to obtain at least one blood pressure reading for the calculation of ankle-brachial index with the failure to feel touch with a 10-g monofilament (OR 1.7, 95% CI 1.3-2.2). Recommendations for annual diabetic foot check in low-risk, community-based patients should be reviewed as absolute events of ulceration are low. The accuracy of foot risk assessment tools to predict ulceration requires evaluation in randomized controlled trials with concurrent economic evaluations.


Neonatal chlordecone exposure impairs early learning and retention of active avoidance in the rat.
The effect of neonatal exposure of rats to chlordecone on the acquisition and retention of active avoidance was investigated. Pups were trained and tested on one-way (preweaning) and/or two-way (post-weaning) active avoidance tasks. Offspring of Fischer-344 rats were administered 1 mg/pup of chlordecone (SC) dissolved in DMSO on postnatal day 4. Body weights were slightly, but significantly, depressed for chlordecone-exposed males (10-11%) and females (7-8%) during preweaning development. Post-weaning body weights were also slightly depressed by the chlordecone treatment (8% for the males, 7% for the females). For pups trained (day 18) on one-way (small to large compartment) active avoidance (OWA), chlordecone treatment increased the number of trials needed to attain the acquisition criterion; the effect was most pronounced in the males. A 72-hr retention test revealed a sex-dependent effect of chlordecone on response latency during the initial test trials. Acquisition of two-way avoidance (TWA) (days 28-30) was superior in female pups relative to males; chlordecone treatment significantly reduced this sex difference in pups which had prior or no prior OWA training. Perhaps most importantly, however, following prior OWA training, vehicle control pups demonstrated a directional bias to make an avoidance response from a small to a large compartment, whereas chlordecone-treated pups executed their avoidance responses in both directions at comparable rates. Similar evidence indicative of a selective retention deficit also characterized TWA performance when a "reversal" procedure was used. A final retention (extinction) session indicated that the chlordecone-treated pups made fewer responses than vehicle-treated controls during the test trials.(ABSTRACT TRUNCATED AT 250 WORDS)


Sclerostin inhibition prevents spinal cord injury-induced cancellous bone loss.
Spinal cord injury (SCI) results in rapid and extensive sublesional bone loss. Sclerostin, an osteocyte-derived glycoprotein that negatively regulates intraskeletal Wnt signaling, is elevated after SCI and may represent a mechanism underlying this excessive bone loss. However, it remains unknown whether pharmacologic sclerostin inhibition ameliorates bone loss subsequent to SCI. Our primary purposes were to determine whether a sclerostin antibody (Scl-Ab) prevents hindlimb cancellous bone loss in a rodent SCI model and to compare the effects of a Scl-Ab to that of testosterone-enanthate (TE), an agent that we have previously shown prevents SCI-induced bone loss. Fifty-five (n = 11-19/group) skeletally mature male Sprague-Dawley rats were randomized to receive: (A) SHAM surgery (T8 laminectomy), (B) moderate-severe (250 kilodyne) SCI, (C) 250 kilodyne SCI + TE (7.0 mg/wk, im), or (D) 250 kilodyne SCI + Scl-Ab (25 mg/kg, twice weekly, sc) for 3 weeks. Twenty-one days post-injury, SCI animals exhibited reduced hindlimb cancellous bone volume at the proximal tibia (via μCT and histomorphometry) and distal femur (via μCT), characterized by reduced trabecular number and thickness. SCI also reduced trabecular connectivity and platelike trabecular structures, indicating diminished structural integrity of the remaining cancellous network, and produced deficits in cortical bone (femoral diaphysis) strength. Scl-Ab and TE both prevented SCI-induced cancellous bone loss, albeit via differing mechanisms. Specifically, Scl-Ab increased osteoblast surface and bone formation, indicating direct bone anabolic effects, whereas TE reduced osteoclast surface with minimal effect on bone formation, indicating antiresorptive effects. The deleterious microarchitectural alterations in the trabecular network were also prevented in SCI + Scl-Ab and SCI + TE animals, whereas only Scl-Ab completely prevented the reduction in cortical bone strength. Our findings provide the first evidence indicating that sclerostin inhibition represents a viable treatment to prevent SCI-induced cancellous and cortical bone deficits and provides preliminary rationale for future clinical trials focused on evaluating whether Scl-Ab prevents osteoporosis in the SCI population.


Epigenetic switching by the metabolism-sensing factors in the generation of orexin neurons from mouse embryonic stem cells.
The orexin system plays a central role in the integration of sleep/wake and feeding behaviors in a broad spectrum of neural-metabolic physiology. Orexin-A and orexin-B are produced by the cleavage of prepro-orexin, which is encoded on the Hcrt gene. To date, methods for generating other peptide neurons could not induce orexin neurons from pluripotent stem cells. Considering that the metabolic status affects orexin expression, we supplemented the culture medium with a nutrient factor, ManNAc, and succeeded in generating functional orexin neurons from mouse ES cells. Because DNA methylation inhibitors and histone deacetylase inhibitors could induce Hcrt expression in mouse ES cells, the epigenetic mechanism may be involved in this orexin neurogenesis. DNA methylation analysis showed the presence of a tissue-dependent differentially methylated region (T-DMR) around the transcription start site of the Hcrt gene. In the orexin neurons induced by supplementation of ManNAc, the T-DMR of the Hcrt gene was hypomethylated in association with higher H3/H4 acetylation. Concomitantly, the histone acetyltransferases p300, CREB-binding protein (CBP), and Mgea5 (also called O-GlcNAcase) were localized to the T-DMR in the orexin neurons. In non-orexin-expressing cells, H3/H4 hypoacetylation and hyper-O-GlcNAc modification were observed at the T-DMRs occupied by O-GlcNAc transferase and Sirt1. Therefore, the results of the present study suggest that the glucose metabolite, ManNAc, induces switching from the inactive state by Ogt-Sirt1 to the active state by Mgea5, p300, and CBP at the Hcrt gene locus.


Ether-Oxygen Containing Electrospun Microfibrous and Sub-Microfibrous Scaffolds Based on Poly(butylene 1,4-cyclohexanedicarboxylate) for Skeletal Muscle Tissue Engineering.
We report the study of novel biodegradable electrospun scaffolds from poly(butylene 1,4-cyclohexandicarboxylate-co-triethylene cyclohexanedicarboxylate) (P(BCE-co-TECE)) as support for in vitro and in vivo muscle tissue regeneration. We demonstrate that chemical composition, i.e., the amount of TECE co-units (constituted of polyethylene glycol-like moieties), and fibre morphology, i.e., aligned microfibrous or sub-microfibrous scaffolds, are crucial in determining the material biocompatibility. Indeed, the presence of ether linkages influences surface wettability, mechanical properties, hydrolytic degradation rate, and density of cell anchoring points of the studied materials. On the other hand, electrospun scaffolds improve cell adhesion, proliferation, and differentiation by favouring cell alignment along fibre direction (fibre morphology), also allowing for better cell infiltration and oxygen and nutrient diffusion (fibre size). Overall, C2C12 myogenic cells highly differentiated into mature myotubes when cultured on microfibres realised with the copolymer richest in TECE co-units (micro-P73 mat). Lastly, when transplanted in the tibialis anterior muscles of healthy, injured, or dystrophic mice, micro-P73 mat appeared highly vascularised, colonised by murine cells and perfectly integrated with host muscles, thus confirming the suitability of P(BCE-co-TECE) scaffolds as substrates for skeletal muscle tissue engineering.


Structural transformation in supercooled water controls the crystallization rate of ice.
One of water's unsolved puzzles is the question of what determines the lowest temperature to which it can be cooled before freezing to ice. The supercooled liquid has been probed experimentally to near the homogeneous nucleation temperature, T(H) ≈ 232 K, yet the mechanism of ice crystallization-including the size and structure of critical nuclei-has not yet been resolved. The heat capacity and compressibility of liquid water anomalously increase on moving into the supercooled region, according to power laws that would diverge (that is, approach infinity) at ~225 K (refs 1, 2), so there may be a link between water's thermodynamic anomalies and the crystallization rate of ice. But probing this link is challenging because fast crystallization prevents experimental studies of the liquid below T(H). And although atomistic studies have captured water crystallization, high computational costs have so far prevented an assessment of the rates and mechanism involved. Here we report coarse-grained molecular simulations with the mW water model in the supercooled regime around T(H) which reveal that a sharp increase in the fraction of four-coordinated molecules in supercooled liquid water explains its anomalous thermodynamics and also controls the rate and mechanisms of ice formation. The results of the simulations and classical nucleation theory using experimental data suggest that the crystallization rate of water reaches a maximum around 225 K, below which ice nuclei form faster than liquid water can equilibrate. This implies a lower limit of metastability of liquid water just below T(H) and well above its glass transition temperature, 136 K. By establishing a relationship between the structural transformation in liquid water and its anomalous thermodynamics and crystallization rate, our findings also provide mechanistic insight into the observed dependence of homogeneous ice nucleation rates on the thermodynamics of water.


The functional annotation of mammalian genomes: the challenge of phenotyping.
The mouse is central to the goal of establishing a comprehensive functional annotation of the mammalian genome that will help elucidate various human disease genes and pathways. The mouse offers a unique combination of attributes, including an extensive genetic toolkit that underpins the creation and analysis of models of human disease. An international effort to generate mutations for every gene in the mouse genome is a first and essential step in this endeavor. However, the greater challenge will be the determination of the phenotype of every mutant. Large-scale phenotyping for genome-wide functional annotation presents numerous scientific, infrastructural, logistical, and informatics challenges. These include the use of standardized approaches to phenotyping procedures for the population of unified databases with comparable data sets. The ultimate goal is a comprehensive database of molecular interventions that allows us to create a framework for biological systems analysis in the mouse on which human biology and disease networks can be revealed.


Mesh tube--constricted varicose veins used as bypass grafts for infrainguinal arterial reconstruction.
Varicose veins are generally deemed inappropriate graft material for arterial reconstructions despite their physiologic flow surface because of their large and irregular caliber. Size reduction by threading such veins in constricting tubes may create bypass grafts of suitable caliber as long as redundant wall material does not cause stenoses. Sixteen human varicose veins (mean +/- SD, 13 +/- 3 mm diameter) obtained after stripping operations were inserted into Dacron mesh tubes of 6 mm internal diameter. Paraffin casts of the distended veins showed a size reduction of 6.9 +/- 2.6 mm. Wall material formed folds in only two veins. In both cases, diameter reduction was more than 10 mm. However, the folds did not result in significant stenoses. Mesh-constricted varicose veins were used as bypass material in 11 infrainguinal arterial reconstructions. All externally supported segments showed satisfactory size reduction without stenoses or folds. One graft occluded 2 months after surgery. Two patients had to undergo reoperation after 2 and 16 months, respectively. None of the complications could be attributed to the constriction of veins. The remaining grafts are patent and functional after a mean of 17 months (range, 6 to 42 months). Considerable size reduction by external wrapping of varicose veins is possible without adverse side effects. Such constricted veins were used successfully as bypass grafts for infrainguinal arterial reconstructions.


Pitfalls in cerebrospinal fluid test for the diagnosis of neurosyphilis.
To determine the usefulness of cerebrospinal fluid tests in the diagnosis of neurosyphilis. Two hundred and seven cerebrospinal fluid-Venereal Disease Research Laboratories tests were performed at King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia between 1992 and 1997. The records of 14 cases with progressive neurological disease and reactive serum fluorescent treponemal absorbent antibodies or treponemal pallidum hemagglutination test were reviewed for clinical presentation, cerebrospinal fluid analysis and Venereal Disease Research Laboratories, neuro-imaging abnormalities and compatibility with the diagnosis of neurosyphilis. The diagnosis of neurosyphilis was made if the patient had reactive serum fluorescent treponemal absorbent antibodies or treponemal pallidum hemagglutination, history of progressive neurological disease and increased cerebrospinal fluid cells or protein. None of the 207 cerebrospinal fluid-Venereal Disease Research Laboratories tests were reactive. The diagnosis of neurosyphilis was made in 10 out of 14 cases with progressive neurological disease and reactive serum rapid plasma reagin, luorescent treponemal absorbent-absorbent antibodies and treponemal pallidum hemagglutination. We conclude that if reactive cerebrospinal fluid-Venereal Disease Research Laboratories is required to confirm or diagnose neurosyphilis, most cases will be overlooked.