Tyrosine kinase and phosphotyrosine phosphatase activity in human promyelocytic leukemia cells and human polymorphonuclear leukocytes.
Although an increase in protein phosphorylation on tyrosine was first noted as a result of cell transformation or the application of growth factors to cells, recent reports have shown high levels of tyrosine kinases in nondividing tissues. For that reason, we have investigated whether normal human polymorphonuclear leukocytes (PMN) contain tyrosine kinase and phosphatase activity. Using a copolymer of glutamine: tyrosine as a substrate for the phosphotransferase reaction, we have demonstrated that PMN contain a cytosolic tyrosine kinase activity that elutes as a single peak from Sephacryl S-200 chromatography and has a molecular weight of 70 kilodaltons. Human promyelocytic leukemia cells (HL-60), contain a similar activity (as demonstrated by column chromatography), with only 25% of the activity found in PMN. This cytosolic tyrosine kinase can phosphorylate angiotensin II and a fragment of the src protein containing tyrosine 416, which suggests a similar substrate specificity to other tyrosine-phosphorylating protein kinases. In addition, we have demonstrated that PMN have double the amount of phosphotyrosine phosphatase (PTPase) activity of that found in HL-60 cells. This enzyme has a Km of 0.932 mmol/L and a Vmax of 0.355 mumol inorganic phosphate released/mg protein/min, which is similar to other cellular PTPase. Activation of PMN with f-Met-Leu-Phe and phorbol esters causes a slight but statistically significant drop in PMN PTPase activity. These results suggest that terminally differentiated myeloid cells have high tyrosine kinase and phosphatase activity, which may play a role in stimulus response coupling in the mature PMN.


Characterization of phiA, a gene essential for phialide development in Aspergillus nidulans.
We have previously identified genes and proteins involved in the fungal response to the Streptomyces-produced antibiotics, bafilomycin B1 and concanamycin A, known inhibitors of V-ATPases. Using mRNA differential display we identified an Aspergillus nidulans gene with 30-fold up-regulated expression in the presence of bafilomycin. This gene, here denoted phiA, and its gene product, were further characterized by targeted gene disruption and immunohistochemistry. Phenotypically, the phiA mutation resulted in reduced growth and severely reduced sporulation. The abnormality could be traced to the phialides, which divided several times instead of forming a single flask-shaped cell. The importance of phiA for phialide and conidium development was supported by immunohistochemistry experiments that showed the protein to be mainly present in these two cell types. Attempts to relate phiA to inhibition of V-ATPases did not result in unambiguous conclusions, but suggest the possibility that changed expression of phiA is correlated with growth arrest caused by inhibited V-ATPases.


Identification of Mycoplasmatales: procedures for both characterization and purification.
Procedures with potential value for identification and purification of Mycoplasma were applied to over 240 cultures representing 11 of the 12 known avian serotypes. Growth was tested at pH 5.5 and 9.5, at 25 and 42 C, in serum-free media, and in the presence of (i) 1% bile salts, (ii) 3% NaCl, and (iii) 0.02% methylene blue. One avian Mycoplasma serotype grew in broth containing 1% bile salts. Two of 11 avian serotypes were resistant to 0.02% methylene blue. A number of Mycoplasma strains grew at 42 C or in a pH 9.5 medium. Usefulness of these procedures for purifying cultures of Mycoplasma is discussed.


ATP-binding cassette transporters in liver.
The human ATP-binding cassette (ABC) superfamily consists of 48 members with 14 of them identified in normal human liver at the protein level. Most of the ABC members act as ATP dependent efflux transport systems. In the liver, ABC transporters are involved in diverse physiological processes including export of cholesterol, bile salts, and metabolic endproducts. Consequently, impaired ABC transporter function is involved in inherited diseases like sitosterolemia, hyperbilirubinemia, or cholestasis. Furthermore, altered expression of some of the hepatic ABCs have been associated with primary liver tumors. This review gives a short overview about the function of hepatic ABCs. Special focus is addressed on the localization and ontogenesis of ABC transporters in the human liver. In addition, their expression pattern in primary liver tumors is discussed.


Differential Placebo Responses for Pharmacotherapy and Neurostimulation in Late-Life Depression.
The magnitude of the placebo response depends on both the modality used as the "placebo" and the intervention with which it is compared, both of which can complicate the interpretation of randomized controlled trials (RCTs) for depression in late life. Given that neurostimulation and pharmacotherapy are among the most common interventions studied for late-life depression, comparing the relative placebo responses in studies of these interventions can aid interpretation of relative effect sizes. We analyzed data from two RCTs of adults aged ≥60 years in an episode of treatment-resistant major depression, one comparing aripiprazole and matching placebo pills and the other comparing deep repetitive transcranial magnetic stimulation (rTMS) and sham rTMS. In both RCTs, depression was assessed using the 17-item Hamilton Depression Rating Scale (HDRS-17). The primary comparison occurred after four weeks using analysis of covariance (ANCOVA) of HDRS-17 scores in participants who received placebo pills or sham rTMS. Relevant covariates included years of education, duration of depressive episode, and baseline HDRS-17 score. Accounting for covariates, there was a larger reduction of HDRS-17 after four weeks in the sham rTMS group (estimated marginal mean ± SE: -5.90 ± 1.45; 95% CI: [-8.82, 2.98]) than in the placebo pills group (-1.07 ± 1.45; [-3.98, 1.85]). There were no significant differences between these groups in the binary outcome analysis of response and remission rates at four weeks or any outcome at trial end point comparison. Sham rTMS may have a larger placebo response than placebo pills early in the treatment of older adults with treatment-resistant depression. Differential placebo responses should be considered in both the interpretation and design of RCTs.


Physics-driven CFD modeling of complex anatomical cardiovascular flows-a TCPC case study.
Recent developments in medical image acquisition combined with the latest advancements in numerical methods for solving the Navier-Stokes equations have created unprecedented opportunities for developing simple and reliable computational fluid dynamics (CFD) tools for meeting patient-specific surgical planning objectives. However, for CFD to reach its full potential and gain the trust and confidence of medical practitioners, physics-driven numerical modeling is required. This study reports on the experience gained from an ongoing integrated CFD modeling effort aimed at developing an advanced numerical simulation tool capable of accurately predicting flow characteristics in an anatomically correct total cavopulmonary connection (TCPC). An anatomical intra-atrial TCPC model is reconstructed from a stack of magnetic resonance (MR) images acquired in vivo. An exact replica of the computational geometry was built using transparent rapid prototyping. Following the same approach as in earlier studies on idealized models, flow structures, pressure drops, and energy losses were assessed both numerically and experimentally, then compared. Numerical studies were performed with both a first-order accurate commercial software and a recently developed, second-order accurate, in-house flow solver. The commercial CFD model could, with reasonable accuracy, capture global flow quantities of interest such as control volume power losses and pressure drops and time-averaged flow patterns. However, for steady inflow conditions, both flow visualization experiments and particle image velocimetry (PIV) measurements revealed unsteady, complex, and highly 3D flow structures, which could not be captured by this numerical model with the available computational resources and additional modeling efforts that are described. Preliminary time-accurate computations with the in-house flow solver were shown to capture for the first time these complex flow features and yielded solutions in good agreement with the experimental observations. Flow fields obtained were similar for the studied total cardiac output range (1-3 1/min); however hydrodynamic power loss increased dramatically with increasing cardiac output, suggesting significant energy demand at exercise conditions. The simulation of cardiovascular flows poses a formidable challenge to even the most advanced CFD tools currently available. A successful prediction requires a two-pronged, physics-based approach, which integrates high-resolution CFD tools and high-resolution laboratory measurements.


Establishment and characterization of a bank of cytotoxic T lymphocytes for immunotherapy of epstein-barr virus-associated diseases.
Adoptive immunotherapy using Epstein-Barr virus (EBV)-specific cytotoxic T lymphocytes (CTL) generated ex vivo can be an effective treatment of EBV-positive posttransplantation lymphoproliferative disease (PTLD). We describe the establishment of a cryopreserved repository of allogeneic virus-specific CTL lines, to our knowledge the first of its kind in the world. CTL lines were grown by weekly stimulation with autologous EBV immortalized lymphoblastoid cell lines (LCLs) from 96 EBV-seropositive blood donors. Analysis of 60 CTL lines grown continuously for 7 to 10 weeks showed an average proportional weekly increase in cell numbers of 1.4, with an overall increase ranging from 1.1 to 83.4. The greatest increase occurred during the early culture period. After four rounds of stimulation, killing of autologous LCLs was generally high (mean 48%); however, most lines required 9 or 10 stimulations to reduce the killing of nonspecific targets. Overall, 79% of CTLs generated showed acceptable levels of specific killing. Phenotypically, the CTL lines consisted of TCRalpha beta+, CD8+ T cells (medians 97% and 90% respectively) with a minority population of CD4+ T cells (median 2%). Most cells expressed the activation and differentiation markers, HLA-DR, CD26, CD45RO, CD69, and CD150. Favorable results have been obtained in an open trial using partially HLA-matched, allogeneic CTLs from this bank to treat PTLD patients. This now represents a single resource that can provide therapeutic CTLs rapidly on a countrywide basis, superseding the time-consuming, expensive practice of generating autologous CTLs from each patient requiring treatment. Additionally, other patient groups, such as those with EBV-positive Hodgkin disease, may benefit from CTL treatment.


Dynamics of repeat-associated plasticity in the aaap gene family in Anaplasma marginale.
Anaplasmosis, the most prevalent tick-transmitted disease of cattle, is caused by the rickettsial intracellular parasite Anaplasma marginale. The pathogen replicates within a parasitophorous vacuole formed from the invagination of the erythrocyte membrane. Several strains of A. marginale form "tails" or "appendages" which are attached to, and extend out from, the cytoplasmic side of the parasitophorous vacuole. Genomic analysis of the parasite antigen distributed along the appendage led to the discovery of the aaap (Anaplasma appendage associated protein) gene family located within a highly plastic region in the genome. The aaap gene family consists of aaap and several alps (for aaap-like proteins), depending on the strain. These genes/proteins are characterized by repeat sequences. To investigate locus plasticity, different versions of the locus were cloned from the same strain as well as from different strains, sequenced and aligned to identify changes. Our findings show that repeat sequences both within and between genes facilitated rearrangement events within the locus. Structural variation of the locus in the St. Maries strain was further investigated during infection of different cellular environments, i.e., bovine erythrocytes and tick cells, with a reduction in subpopulations of the aaap locus within the tick as compared to erythrocytes. Interestingly, subpopulations bearing alternative locus structures began to arise again when the pathogen was transferred from the tick environment into a naïve calf. Additionally, the Aaap protein expression profile between blood and tick samples showed a regulatory shift, indicating a host-specific response. Alignment of the protein sequences from different species of Anaplasma reveals six similar repeating motifs that appear to be unique to a few species of Anaplasma. The role the aaap locus may play in the pathogenesis of the bovine host or in tick infection/transmission remains unknown; however, the changes in aaap locus subpopulations, locus structure, and protein expression indicate that these genes have a role in strain diversification.


Inhaled fluticasone causes iatrogenic cushing's syndrome in patients treated with Ritonavir.
Ritonavir, a protease inhibitor (PI), is commonly used in the treatment of HIV-1 infection. It is a potent inhibitor of the hepatic cytochrome P450 superfamily. Therefore, its usage with other PI medications leads to significant increases in the levels of the latter PI, which allows a reduction in pill burden. Intranasal and inhaled corticosteroids are widely used for the treatment of allergic rhinitis and asthma. Inhaled steroids do not usually lead to systemic adverse events, since their plasma concentrations are quite low due to extensive first-pass metabolism and clearance by CYP3A4. However, the coadministration of Ritonavir with inhaled (or intranasal) corticosteroids may result in an increase in the plasma corticosteroid levels due to the potent CYP3A4 inhibition by Ritonavir. This may cause Cushing's syndrome (laboratory and clinical) with adrenal suppression. Plasma cortisol and urinary-free cortisol levels were determined using immunoassays. In the Synacthen test, plasma cortisol levels were measured at time 0 as well as at times 60, 120, and 150 minutes following an intramuscular injection of 0.25 mg Synacthen. We present here three HIV-1 female patients aged 12, 55 and 65 years who developed iatrogenic Cushing's syndrome with adrenal suppression following the coadministration of Ritonavir and inhaled Fluticasone, both at the standard recommended doses. The coadministration of Ritonavir and Fluticasone at the recommended doses caused, in our three patients, iatrogenic Cushing's syndrome with adrenal suppression. We suggest that this adverse event is underdiagnosed and high clinical suspicion is needed for early diagnosis and prenention of Addisonian crises. Thus, Fluticasone treatment should be avoided in patients who are treated with Ritonavir. Alternative therapeutic options for asthma control such as oral Montelukast or bronchodilators alone should be considered.


Electric Double Layer Composed of an Antagonistic Salt in an Aqueous Mixture: Local Charge Separation and Surface Phase Transition.
We examine an electric double layer containing an antagonistic salt in an aqueous mixture, where the cations are small and hydrophilic but the anions are large and hydrophobic. In this situation, a strong coupling arises between the charge density and the solvent composition. As a result, the anions are trapped in an oil-rich adsorption layer on a hydrophobic wall. We then vary the surface charge density σ on the wall. For σ>0 the anions remain accumulated, but for σ<0 the cations are attracted to the wall with increasing |σ|. Furthermore, the electric potential drop Ψ(σ) is nonmonotonic when the solvent interaction parameter χ(T) exceeds a critical value χ_{c} determined by the composition and the ion density in the bulk. This leads to a first-order phase transition between two kinds of electric double layers with different σ and common Ψ. In equilibrium such two-layer regions can coexist. The steric effect due to finite ion sizes is crucial in these phenomena.


The Diagnosis of Feline Leukaemia Virus (FeLV) Infection in Owned and Group-Housed Rescue Cats in Australia.
A field study was undertaken to (i) measure the prevalence of feline leukaemia virus (FeLV) exposure and FeLV infection in a cross-section of healthy Australian pet cats; and (ii) investigate the outcomes following natural FeLV exposure in two Australian rescue facilities. Group 1 (n = 440) consisted of healthy client-owned cats with outdoor access, predominantly from eastern Australia. Groups 2 (n = 38) and 3 (n = 51) consisted of a mixture of healthy and sick cats, group-housed in two separate rescue facilities in Sydney, Australia, tested following identification of index cases of FeLV infection in cats sourced from these facilities. Diagnostic testing for FeLV exposure/infection included p27 antigen testing using three different point-of-care FeLV kits and a laboratory-based ELISA, real-time polymerase chain reaction (qPCR) testing to detect FeLV proviral DNA in leukocytes, real-time reverse-transcription PCR (qRT-PCR) testing to detect FeLV RNA in plasma, and neutralising antibody (NAb) testing. Cats were classified as FeLV-uninfected (FeLV-unexposed and presumptively FeLV-abortive infections) or FeLV-infected (presumptively regressive and presumptively progressive infections). In Group 1, 370 FeLV-unexposed cats (370/440, 84%), 47 abortive infections (47/440, 11%), nine regressive infections (9/440, 2%), and two progressive infections (2/440, 0.5%) were identified, and 12 FeLV-uninfected cats (12/440, 3%) were unclassifiable as FeLV-unexposed or abortive infections due to insufficient samples available for NAb testing. In Groups 2 and 3, 31 FeLV-unexposed cats (31/89, 35%), eight abortive infections (8/89, 9%), 22 regressive infections (22/89; 25%), and 19 progressive infections (19/89; 21%) were discovered, and nine FeLV-uninfected cats (9/89; 10%) were unclassifiable due to insufficient samples available for NAb testing. One of the presumptively progressively-infected cats in Group 3 was likely a focal FeLV infection. Two other presumptively progressively-infected cats in Group 3 may have been classified as regressive infections with repeated testing, highlighting the difficulties associated with FeLV diagnosis when sampling cats at a single time point, even with results from a panel of FeLV tests. These results serve as a reminder to Australian veterinarians that the threat of FeLV to the general pet cat population remains high, thus vigilant FeLV testing, separate housing for FeLV-infected cats, and FeLV vaccination of at-risk cats is important, particularly in group-housed cats in shelters and rescue facilities, where outbreaks of FeLV infection can occur.


Treatment of a mandibular canine abutment with two canals.
Human mandibular canines do not present an internal anatomy as simple as could be expected; there are such canines with a single root and two canals, two roots or fused roots. The existence of mandibular canines with more than one root canal is a fact that clinicians ought to keep in mind, in order to avoid failure during endodontic treatment. In spite of the low incidence of lower canines with one root and two canals, this possibility cannot be forgotten, inasmuch as the presence of a second canal in these teeth leads to difficulties in endodontic treatment. The precise knowledge of the dental endocanalicular system's anatomy is essential in the success of the root canal therapy, because the failure to detect the accessories canals and the incomplete radicular obturation leads to the infection of the periapical space, which will ultimately result in the loss of the tooth. The long-term success of the prosthodontic treatment depends directly on the quality of the endodontic treatment realized on the abutment teeth. The mandibular canine is very important as abutment for any type of prosthetic restoration. This article presents a clinical case of a canine tooth which displays a radicular morphology with two canals, which leads us to conclude that such anatomical variations on human teeth can also occur in our country as much as described in the international literature and cannot be overlooked when treating the teeth.


Characterization of the insertion of Pseudomonas exotoxin A into membranes.
Pseudomonas aeruginosa exotoxin A (PTx) is an extremely potent inhibitor of protein synthesis, similar to diphtheria toxin in its mode of action. It is synthesized in precursor form and secreted as an Mr 66,583 protein lacking a 25-amino acid leader sequence. While the primary sequence and the nature of the enzyme activity that leads to inactivation of elongation factor 2 are known, the mechanism of PTx internalization remains obscure. To elucidate the entry pathway, we examined PTx-membrane interactions using vesicle targets of defined lipid composition. Insertion was monitored with an intramembranous photoreactive probe; pore formation was determined from liposomal swelling rates. Our results show that the efficiency of PTx binding to vesicles increases dramatically with decreasing pH. In general, the insertion efficiency correlated with the binding efficiency. At pH 4, we noted a slight decrease in binding below the melting point (23 degrees C) of the target vesicles. Not only was PTx able to insert into frozen bilayers, but the efficiency of penetration at 0 degrees C was actually somewhat higher than expected based on binding efficiency. Liposome swelling assays analyzed by the Renkin equations indicated that PTx-liposomes made at pH 4 were permeable to solutes up to 2.8 nm in diameter. Pores of a similar size were found when the liposomes were made at pH 7, but the efficiency of pore formation at this pH was very low. Chymotrypsin fragmentation profiles of PTx depended on incubation conditions, e.g., pH, presence of NAD, reducing agents, and membranes. Liposomes containing PTx cleaved at pH 4 displayed up to 40-fold more pore activity than liposomes containing uncleaved PTx or PTx cleaved at pH 7. Pore activity at pH 7 was negligible. Addition of reducing agents caused a 50 to 60% increase in pore activity. Cleaved toxin was active in target membrane insertion even at 0 degrees C, and all of the major fragments were photolabeled.


Effect of different levels of rapidly degradable carbohydrates calculated by a simple rumen model on performance of lactating dairy cows.
Aggregating rumen degradation characteristics of different carbohydrate components into the term modeled rapidly degradable carbohydrates (mRDC) can simplify diet formulation by accounting for differences in rate and extent of carbohydrate degradation within and between feedstuffs. This study sought to evaluate responses of lactating dairy cows to diets formulated with increasing levels of mRDC, keeping the supply of other nutrients as constant as possible. The mRDC content of feedstuffs was calculated based on a simple rumen model including soluble, washable, and nonwashable but potentially degradable fractions, as well as the fractional degradation and passage rates, of sugar, starch, neutral detergent fiber, and other carbohydrates. The mRDC term effectively represents the total amount of carbohydrates degraded in the rumen within 2 h after ingestion. Fifty-two lactating Holstein cows (of which 4 were rumen fistulated) were assigned to 4 treatments in a 4 × 4 Latin square design. Treatments were fed as a total mixed ration consisting of 25.4% corn silage, 23.1% grass silage, 11.6% grass hay, and 39.9% concentrate on a dry matter basis. Differences in mRDC were created by exchanging nonforage neutral detergent fiber-rich ingredients (mainly sugar beet pulp) with starch-rich ingredients (mainly wheat) and by exchanging corn (slowly degradable starch) with wheat (rapidly degradable starch) in the concentrate, resulting in 4 treatments that varied in dietary mRDC level of 167, 181, 194, or 208 g/kg of dry matter. Level of mRDC did not affect dry matter intake. Fat- and protein-corrected milk production and milk fat and lactose yield were greatest at 181 mRDC and decreased with further increases in mRDC. Milk protein yield and concentration increased with increasing mRDC level. Mean rumen pH and diurnal variation in ruminal pH did not differ between treatments. Total daily meal time and number of visits per meal were smaller at 181 and 194 mRDC. Despite milk production responses, increasing dietary mRDC levels, while maintaining net energy and intestinal digestible protein as well as other nutrients at similar levels, did not influence rumen pH parameter estimates and had minor effects on feeding behavior. These results indicate that aggregating rapidly degradable carbohydrate content into one term may be a simple way to further improve predictability of production responses in practical diet formulation for lactating dairy cows.


Neural circuits mediate electrosensory behavior in Caenorhabditis elegans.
The nematode Caenorhabditis elegans deliberately crawls toward the negative pole in an electric field. By quantifying the movements of individual worms navigating electric fields, we show that C. elegans prefers to crawl at specific angles to the direction of the electric field in persistent periods of forward movement and that the preferred angle is proportional to field strength. C. elegans reorients itself in response to time-varying electric fields by using sudden turns and reversals, standard reorientation maneuvers that C. elegans uses during other modes of motile behavior. Mutation or laser ablation that disrupts the structure and function of amphid sensory neurons also disrupts electrosensory behavior. By imaging intracellular calcium dynamics among the amphid sensory neurons of immobilized worms, we show that specific amphid sensory neurons are sensitive to the direction and strength of electric fields. We extend our analysis to the motor level by showing that specific interneurons affect the utilization of sudden turns and reversals during electrosensory steering. Thus, electrosensory behavior may be used as a model system for understanding how sensory inputs are transformed into motor outputs by the C. elegans nervous system.


Molecular epidemiology of hepatitis E virus in humans, pigs and wild boars in Sweden.
Hepatitis E infections in humans are usually acquired in endemic countries in Asia or Africa. In Sweden 17 cases infected in Europe, between 1993 and 2009, were identified. All had clinical hepatitis E with unknown source of infection. Hepatitis E virus (HEV) was identified in faecal samples from 63 piglets in 12 pig farms in Sweden. HEV was also identified in blood from 13 out of 159 investigated Swedish wild boars from nine counties. Partial HEV genomes from humans, pigs and wild boars were sequenced and compared by phylogeny. The results showed close relatedness between HEV strains from piglets from the same farm and from wild boars from the same county. HEV strains from humans showed relatedness with strains from pigs and wild boars from the same county. This study showed that HEV strains form geographical clusters in the phylogenetic tree. The methods used in this study may thus be used for tracing the origin of an infecting strain. Furthermore, this study indicated that there are endemic sources of human HEV infections in Sweden.


Hepatitis B virus infection and hepatocellular carcinoma among parous Taiwanese women: nationwide cohort study.
Few long-term studies of hepatitis B virus (HBV) infection and hepatocellular carcinoma have focused on women. We used a nationwide cohort of reproductive-aged Taiwanese women to study relationships of HBV infection and parity with hepatocellular carcinoma risk. Prenatal test results were available for hepatitis B surface antigen (HBsAg) and e antigen (HBeAg) in the National Hepatitis B Vaccination Registry from 1 782 401 pregnant women tested from October 1, 1983, through March 31, 2000. Data from the 306 women who were diagnosed with hepatocellular carcinoma were ascertained during 15 901 722 person-years of follow-up through linkage with National Cancer Registry and National Death Certification Registry. We used Cox proportional hazards models to investigate the association of age and reproductive and serological parameters with the risk of hepatocellular carcinoma. Incidence rates for hepatocellular carcinoma were 0.55, 7.91, and 8.76 per 100 000 person-years among women who were HBsAg negative (ie, noncarriers), HBsAg positive plus HBeAg negative, and HBsAg positive plus HBeAg positive, respectively (compared with noncarriers, for HBsAg-positive and HBeAg-positive women, age-adjusted hazard ratio [HR] for developing hepatocellular carcinoma = 17.31, 95% confidence interval [CI] = 12.08 to 24.81; and for HBsAg-negative plus HBeAg-positive women, HR = 13.94, 95% CI = 10.34 to 18.79). Incidence rates were 0.39, 3.10, and 9.01 per 100 000 person-years, respectively, among persistent noncarriers, HBsAg-serocleared carriers, and persistent HBsAg carriers (compared with noncarriers, for HBsAg-serocleared carriers, age-adjusted HR = 7.95, 95% CI = 3.50 to 18.04; and for HBsAg persistence, HR = 23.13, 95% CI = 14.23 to 37.61). Incidence rates were 2.04, 1.55, and 1.66 per 100 000 person-years for women who had one, two, or three or more children, respectively (compared with one child, for two children, age- and HBsAg-adjusted HR = 0.68, 95% CI = 0.50 to 0.93; and for three or more children, HR = 0.63, 95% CI = 0.42 to 0.92). The risk for hepatocellular carcinoma was statistically significantly higher among women with chronic or active HBV infections and among those with persistent HBV infection or who underwent HBsAg seroclearance during follow-up than among HBV-unexposed women. This risk decreased as parity increased, independent of HBsAg status and age.


[Effects of chronic irradiation on the adaptive potential of plants].
The content of anthocianins was determined in Oenothera biennis plants, grown from seeds, picked on the plots with different levels of radionuclide contamination in the 30-km Chernobyl zone. It was shown that the content of anthocianins was higher in plants from Yanov area (20-40 mR/h) than in plants from Chernobyl area (0.04 mR/h). An acute gamma-irradiation of seeds with a dose of 5-100 Gy or UV-irradiation of plants resulted in increasing of anthocianin content which was higher in plants grown from the seeds picked on plots with a low level of radionuclide contamination. The data obtained suggest that chronic irradiation of O. biennis populations induces accumulation of anthocianins. Apparently the adaptivity potential has been more completely realised in plants on the plots with a higher level of radionuclide contamination. The populations which were formed in the absence or at the low level of radionuclide contamination, on the contrary, have a significant adaptivity potential and, accordingly, higher radioresistance.


Should we use a direct regulation to implement the Healthy Prisons Agenda in England? A qualitative study among prison key policy makers.
The Healthy Prisons Agenda seeks to reduce prisoners' health risks, balance prisoners' rights with a security regime, ensure equivalent prison health service provisions to community health services, and facilitate the whole-prison approach. There is an established assumption that legislation will ensure better implementation of health promotion programmes. This study aimed to examine whether a legislative framework, via a direct regulation, could lead to enhanced implementation of the Healthy Prisons Agenda in England. A qualitative study design was conducted using semi-structured interviews with 30 key prison policy makers in England. Our findings contradict the established assumption that legislation improves the implementation of health promotion programmes. A direct regulation was perceived as restrictive, manifesting excessive compliance and encouraging a risk-averse culture, whilst preoccupation with security, order and discipline amongst prison governors and custody staff was deemed an internal institutional barrier to implementing the Healthy Prisons Agenda. External barriers included diminishing resources, lengthier or delayed sentencing, and an unsympathetic public and political stance towards prisoner rehabilitation. A direct regulation should not be used to operationalize the Healthy Prisons Agenda. Rather, self-regulation, along with proactive solutions for the identified barriers to implementing the Agenda, is the most appropriate path forward.


Deformable image registration for radiation therapy: principle, methods, applications and evaluation.
Background: Deformable image registration (DIR) is increasingly used in the field of radiation therapy (RT) to account for anatomical deformations. The aims of this paper are to describe the main applications of DIR in RT and discuss current DIR evaluation methods. Methods: Articles on DIR published from January 2000 to October 2018 were extracted from PubMed and Science Direct. Our search was restricted to articles that report data obtained from humans, were written in English, and address DIR methods for RT. A total of 207 articles were selected from among 2506 identified in the search process. Results: At planning, DIR is used for organ delineation using atlas-based segmentation, deformation-based planning target volume definition, functional planning and magnetic resonance imaging-based dose calculation. In image-guided RT, DIR is used for contour propagation and dose calculation on per-treatment imaging. DIR is also used to determine the accumulated dose from fraction to fraction in external beam RT and brachytherapy, both for dose reporting and adaptive RT. In the case of re-irradiation, DIR can be used to estimate the cumulated dose of the two irradiations. Finally, DIR can be used to predict toxicity in voxel-wise population analysis. However, the evaluation of DIR remains an open issue, especially when dealing with complex cases such as the disappearance of matter. To quantify DIR uncertainties, most evaluation methods are limited to geometry-based metrics. Software companies have now integrated DIR tools into treatment planning systems for clinical use, such as contour propagation and fraction dose accumulation. Conclusions: DIR is increasingly important in RT applications, from planning to toxicity prediction. DIR is routinely used to reduce the workload of contour propagation. However, its use for complex dosimetric applications must be carefully evaluated by combining quantitative and qualitative analyses.


Outbreak investigation and control case report of brucellosis: experience from livestock research centre, Mpwapwa, Tanzania.
Brucellosis screening was conducted between 2005 and 2010 at the National Livestock Research Institute headquarters, Mpwapwa, Tanzania, following an abortion storm in cattle. The initial screening targeted breeding herds; 483 cattle were screened using the Rose Bengal Plate Test (RBPT) followed by the Competitive Enzyme-linked Immunosorbent Assay (c-ELISA) as a confirmatory test. The seropositivity on c-ELISA was 28.95% in 2005; it subsequently declined to 6.72%, 1.17%, 0.16% and 0.00% in 2006, 2007, 2009 and 2010, respectively. Brucella seropositivity was not detected in goats. Seropositivity declined following institution of stringent control measures that included: gradual culling of seropositive animals through slaughter; isolation and confinement of pregnant cows close to calving; proper disposal of placentas and aborted foetuses; the use of the S19 vaccine; and restricted introduction of new animals. It was thought that the source of this outbreak was likely to have been from the introduction of infected animals from another farm. Furthermore, humans were found with brucellosis antibodies. Out of 120 people screened, 12 (10%) were confirmed seropositive to brucella antigen exposure by c-ELISA analysis. The majority of the seropositive individuals (80%) were milkers and animal handlers from the farm. Nine individuals had clinical signs suggestive of brucellosis. All cases received medical attention from the district hospital. This achievement in livestock and human health showed that it is possible to control brucellosis in dairy farms, compared to pastoral and agro-pastoral farms, thus providing evidence to adopt these strategies in dairy farms thought to be at risk.


Transillumination to evaluate spermatogenesis: effect of testosterone enanthate in adult male rats.
The present study has been designed to determine which stages of seminiferous cycle were modified by in vivo intramuscular administration of one or two doses of 25 mg of testosterone enanthate. Testicular weight was significantly reduced after treatment with two doses of testosterone enanthate (p < .05). Assessment of total lengths of stages per 100 mm of seminiferous tubules scored by transillumination showed that testosterone enanthate increased significantly the length of stages VI-VII, but it reduced total lengths of stages I, XII, and XIII-XIV. A dose-dependent reduction in the absolute and relative frequency of stage I was observed with testosterone enanthate treatment, whereas stages VI-VII were significantly increased. Analysis of relative frequencies of stages showed that stages XIII-XIV and stages I-V were significantly below the control value (1), whereas stages VI-VIII were over 1. In summary, results of the present study showed that transillumination is an adequate technique to assess the effect of drugs on spermatogenesis, and that testosterone enanthate modifies the pattern of the stages of the seminiferous epithelium cycle, before testicular weight is affected.


Development of a colorimetric microfluidic pH sensor for autonomous seawater measurements.
High quality carbonate chemistry measurements are required in order to fully understand the dynamics of the oceanic carbonate system. Seawater pH data with good spatial and temporal coverage are particularly critical to apprehend ocean acidification phenomena and their consequences. There is a growing need for autonomous in situ instruments that measure pH on remote platforms. Our aim is to develop an accurate and precise autonomous in situ pH sensor for long term deployment on remote platforms. The widely used spectrophotometric pH technique is capable of the required high-quality measurements. We report a key step towards the miniaturization of a colorimetric pH sensor with the successful implementation of a simple microfluidic design with low reagent consumption. The system is particularly adapted to shipboard deployment: high quality data was obtained over a period of more than a month during a shipboard deployment in northwest European shelf waters, and less than 30 mL of indicator was consumed. The system featured a short term precision of 0.001 pH (n=20) and an accuracy within the range of a certified Tris buffer (0.004 pH). The quality of the pH system measurements have been checked using various approaches: measurements of certified Tris buffer, measurement of certified seawater for DIC and TA, comparison of measured pH against calculated pH from pCO2, DIC and TA during the cruise in northwest European shelf waters. All showed that our measurements were of high quality. The measurements were made close to in situ temperature (+0.2°C) in a sampling chamber which had a continuous flow of the ship's underway seawater supply. The optical set up was robust and relatively small due to the use of an USB mini-spectrometer, a custom made polymeric flow cell and an LED light source. The use of a three wavelength LED with detection that integrated power across the whole of each LED output spectrum indicated that low wavelength resolution detectors can be used instead of the current USB mini spectrophotometer. Artefacts due to the polychromatic light source and inhomogeneity in the absorption cell are shown to have a negligible impact on the data quality. The next step in the miniaturization of the sensor will be the incorporation of a photodiode as detector to replace the spectrophotometer.


Study of recovery model implementation in psychiatric hospitals: Clients and multidisciplinary staff perspective.
The recovery model guides mental health services. However, the delivery of recovery-oriented services in inpatient settings is still a challenge. Factors affecting recovery model implementation can be classified into three types: the hospital environment, the inpatient and the service provider. This study aims to quantitatively evaluate the impact of environment, inpatient and service provider factors on recovery model implementation in hospitals. Forty-five service providers and 42 inpatients from three types of wards (acute locked, acute open and daycare) of two hospitals participated in this cross-sectional study. We assessed inpatient cognition, functional capacity and illness severity. In addition, we retrieved information on service providers' professional status and evaluated the recovery model knowledge and attitudes. Implementation of the recovery model was measured using the Recovery Self-Assessment, both the inpatient and service provider versions. Differences were found between the three types of wards in recovery orientation as reported by service providers ( χ 2 2 = 15.3, P < 0.001), but not by inpatients ( χ 2 2 = 2.34, P > 0.05). Providers' internalized knowledge and attitudes toward recovery, inpatients' functional capacity and age of illness onset were associated with recovery implementation (0.31 < r < 0.48, P < 0.05). The findings confirm quantitatively the multilevel nature of factors that affect the implementation of the recovery model in psychiatric hospitals. The inpatients' perspective should be incorporated into the service development process. Based on the study results, the reduction in the restrictive features of the wards' environment is recommended. Promotion of the recovery model implementation in the hospital setting requires the expansion of staff's internal positive attitudes toward recovery.


Controlling malaria using livestock-based interventions: a one health approach.
Where malaria is transmitted by zoophilic vectors, two types of malaria control strategies have been proposed based on animals: using livestock to divert vector biting from people (zooprophylaxis) or as baits to attract vectors to insecticide sources (insecticide-treated livestock). Opposing findings have been obtained on malaria zooprophylaxis, and despite the success of an insecticide-treated livestock trial in Pakistan, where malaria vectors are highly zoophilic, its effectiveness is yet to be formally tested in Africa where vectors are more anthropophilic. This study aims to clarify the different effects of livestock on malaria and to understand under what circumstances livestock-based interventions could play a role in malaria control programmes. This was explored by developing a mathematical model and combining it with data from Pakistan and Ethiopia. Consistent with previous work, a zooprophylactic effect of untreated livestock is predicted in two situations: if vector population density does not increase with livestock introduction, or if livestock numbers and availability to vectors are sufficiently high such that the increase in vector density is counteracted by the diversion of bites from humans to animals. Although, as expected, insecticide-treatment of livestock is predicted to be more beneficial in settings with highly zoophilic vectors, like South Asia, we find that the intervention could also considerably decrease malaria transmission in regions with more anthropophilic vectors, like Anopheles arabiensis in Africa, under specific circumstances: high treatment coverage of the livestock population, using a product with stronger or longer lasting insecticidal effect than in the Pakistan trial, and with small (ideally null) repellency effect, or if increasing the attractiveness of treated livestock to malaria vectors. The results suggest these are the most appropriate conditions for field testing insecticide-treated livestock in an Africa region with moderately zoophilic vectors, where this intervention could contribute to the integrated control of malaria and livestock diseases.


[Prevalence of Dental Fluorosis and Caries in Mexico City Schoolchildren].
This study was conducted to determine prevalence and severity of dental fluorosis and dental caries in school-children living in two zones of the Mexico City's East Area. Fluorosis and caries scores were recorded for 1,569 children according to the Dean Community Index, DMFT and DMFS indexes, and the relationship between fluorosis and fluoride sources was investigated. Dental fluorosis was found in 60.42% of children. Dean's Community score was 0.96 (SD-0.58). Quantity of toothpaste employed, toothbrushing frequency, and early initiation of toothbrushing correlated well with fluorosis (p = 0.03). Caries was present in 70.52% of children, DMFT and DMFS scores were 2.64 (SD 2.4) and 3.97 (SD 4.18) respectively. DMFT and DMFS scores were lower in children with mild and moderate fluorosis than in those fluorosis-free children. (p = 0.03).


Effects of L-carnitine and niacin supplied by drinking water on fattening performance, carcass quality and plasma L-carnitine concentration of broiler chicks.
The present study was initiated to determine whether dietary supplemental L-carnitine and niacin affect growth performance, carcass yield, abdominal fat and plasma L-carnitine concentration of broiler chicks. One-day-old broiler chicks (COB500) were used in the experiment. A two by two factorial arrangement was employed with two levels (0 and 50 mg/l) of supplemental L-carnitine and two levels (0 or 50 mg/l) of supplemental niacin in drinking water as main effects. Body weight gain was significantly improved by L-carnitine, or L-carnitine + niacin supplementation during the first 3 weeks. However, supplemental L-carnitine and niacin did not change body weight gain during the last 3 weeks of the experimental period. Supplemental L-carnitine significantly improved feed intake during the first 3 weeks. Supplemental L-carnitine or niacin did not influence carcass weight, carcass yield and abdominal fat weight. L-carnitine content in the plasma was significantly higher in the groups receiving supplemental L-carnitine and L-carnitine + niacin. It is concluded that dietary supplemental L-carnitine or L-carnitine + niacin could have positive effects on body weight gain and feed intake during the early stages of growing. However, supplemental L-carnitine or L-carnitine + niacin were not of benefit regarding the complete growth period.


Effect of silymarin (milk thistle) on liver disease in patients with chronic hepatitis C unsuccessfully treated with interferon therapy: a randomized controlled trial.
The botanical product silymarin, an extract of milk thistle, is commonly used by patients to treat chronic liver disease, despite scant and conflicting evidence of its efficacy. To determine the effect of silymarin on liver disease activity in patients with chronic hepatitis C virus (HCV) infection unsuccessfully treated with interferon-based therapy. Multicenter, double-blind, placebo-controlled trial conducted at 4 medical centers in the United States. Participants included 154 persons with chronic HCV infection and serum alanine aminotransferase (ALT) levels of 65 U/L or greater who were previously unsuccessfully treated with interferon-based therapy. Enrollment began in May 2008 and was completed in May 2010, with the last follow-up visit completed in March 2011. Participants were randomly assigned to receive 420-mg silymarin, 700-mg silymarin, or matching placebo administered 3 times per day for 24 weeks. The primary outcome measure was serum ALT level of 45 U/L or less (considered within the normal range) or less than 65 U/L, provided this was at least a 50% decline from baseline values. Secondary outcomes included changes in ALT levels, HCV RNA levels, and quality-of-life measures. After 24 weeks of treatment, only 2 participants in each treatment group (P ≥ .99) met the primary outcome measure (3.8% [95% CI, 0.5% to 13.2%] for placebo, 4.0% [95% CI, 0.5% to 13.7%] for 420-mg silymarin, and 3.8% [95% CI, 0.5% to 13.2%] for 700-mg silymarin). The mean decline in serum ALT activity at the end of treatment did not differ significantly (P = .75) across the 3 treatment groups (mean decline, -4.3 [95% CI, -17.3 to 8.7] U/L for placebo, -14.4 [95% CI, -41.6 to 12.7] U/L for 420-mg silymarin, -11.3 [95% CI, -27.9 to 5.4] U/L for 700-mg silymarin); there likewise were no significant differences in HCV RNA levels (mean change, 0.07 [95% CI, -0.05 to 0.18] log10 IU/mL for placebo, -0.03 [95% CI, -0.18 to 0.12] log10 IU/mL for 420-mg silymarin, 0.04 [95% CI, -0.08 to 0.16] log10 IU/mL for 700-mg silymarin; P = .54) or quality-of-life measures. The adverse event profile of silymarin was comparable with that of placebo. Higher than customary doses of silymarin did not significantly reduce serum ALT levels more than placebo in participants with chronic HCV infection unsuccessfully treated with interferon-based therapy. clinicaltrials.gov Identifier: NCT00680342.


Cultured blood dendritic cells retain HIV-1 antigen-presenting capacity for memory CTL during progressive HIV-1 infection.
Dendritic cells (DC) are potent APC that may be involved in the pathogenesis of HIV-1 infection. We studied the APC function of DC from HIV-1-infected subjects that were derived from monocyte-depleted PBMC by culture in human IL-4 and human granulocyte-macrophage CSF. The cultured cells from the HIV-1-infected subjects had similar morphology and phenotype of mature DC (CD80 = 41 +/- 8%, CD86 = 77 +/- 5%, CD40 = 87 +/- 6%, CD1a = 1 +/- 1%) to DC cultured from seronegative subjects. The yield of these DC was lower than from HIV-1-seronegative subjects (4 +/- 0% vs 11 +/- 2%, p < 0.01), and the lower DC yields correlated with lower numbers of blood CD4+ T cells (r = 0.60, p < 0.01) and higher plasma viral load (r = -0.49, p < 0.01). DC from HIV-1-infected subjects were infected with recombinant vaccinia virus vectors expressing Gag, Pol, and Env and were able to stimulate equal or higher levels of MHC class I-restricted, anti-HIV-1 memory CTL (CTLm) than were similarly treated, autologous B lymphocyte cell lines. DC pulsed with peptides representing HIV-1 CTL epitopes stimulated higher levels of anti-HIV-1 CTLm responses than did DC infected with the vaccinia virus-HIV-1 constructs. Allogeneic, MHC class I-matched DC also stimulated anti-HIV-1 CTLm activity in cells from HIV-1-infected subjects. DC from early and late stages of HIV-1 infection had a similar ability to activate CTLm specific for targets expressing either HIV-1 genes via vaccinia virus vectors or HIV-1 immunodominant synthetic peptides. However, DC from either early or late stages of HIV-1 infection could not overcome the defect in anti-HIV-1 CTLm response in advanced infection.


Leukocyte function-associated antigen 1-mediated adhesion stability is dynamically regulated through affinity and valency during bond formation with intercellular adhesion molecule-1.
Neutrophil rolling and transition to arrest on inflamed endothelium are dynamically regulated by the affinity of the beta(2) integrin CD11a/CD18 (leukocyte function associated antigen 1 (LFA-1)) for binding intercellular adhesion molecule (ICAM)-1. Conformational shifts are thought to regulate molecular affinity and adhesion stability. Also critical to adhesion efficiency is membrane redistribution of active LFA-1 into dense submicron clusters where multimeric interactions occur. We examined the influences of affinity and dimerization of LFA-1 on LFA-1/ICAM-1 binding by engineering a cell-free model in which two recombinant LFA-1 heterodimers are bound to respective Fab domains of an antibody attached to latex microspheres. Binding of monomeric and dimeric ICAM-1 to dimeric LFA-1 was measured in real time by fluorescence flow cytometry. ICAM-1 dissociation kinetics were measured while LFA-1 affinity was dynamically shifted by the addition of allosteric small molecules. High affinity LFA-1 dissociated 10-fold faster when bound to monomeric compared with dimeric ICAM-1, corresponding to bond lifetimes of 25 and 330 s, respectively. Downshifting LFA-1 into an intermediate affinity state with the small molecule I domain allosteric inhibitor IC487475 decreased the difference in dissociation rates between monomeric and dimeric ICAM-1 to 4-fold. When LFA-1 was shifted into the low affinity state by lovastatin, both monomeric and dimeric ICAM-1 dissociated in less than 1 s, and the dissociation rates were within 50% of each other. These data reveal the respective importance of LFA-1 affinity and proximity in tuning bond lifetime with ICAM-1 and demonstrate a nonlinear increase in the bond lifetime of the dimer versus the monomer at higher affinity.


A cloned frog vasoactive intestinal polypeptide/pituitary adenylate cyclase-activating polypeptide receptor exhibits pharmacological and tissue distribution characteristics of both VPAC1 and VPAC2 receptors in mammals.
Three receptor subtypes for the neuropeptides vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) have been identified in mammals: the PAC1 receptor (PAC1-R) which is selectively activated by PACAP, and two VPAC receptors (VPAC1-R and VPAC2-R), which are equally stimulated by PACAP and VIP. The structures of PACAP and VIP have been well conserved during evolution, but little is known about VIP/PACAP receptors in nonmammalian species. An amphibian VIP/PACAP receptor complementary DNA (cDNA) has been cloned and characterized from a frog (Rana ridibunda) pituitary cDNA library. The predicted protein contains seven putative transmembrane domains and exhibits the highest sequence identity (65%) with the human VPAC1-R. The cloned cDNA was transiently expressed in LLC-PK1 cells, and its pharmacological profile was determined in comparison with the human VPAC1-R. Both PACAP and VIP stimulated cAMP accumulation through the cloned receptor with an EC50 of about 30 nM. In contrast, secretin, at concentrations that stimulate the human VPAC1-R, had no effect on cAMP production. RT-PCR analysis revealed the widespread distribution of this frog VIP/PACAP receptor in peripheral tissues. In situ hybridization histochemistry using a complementary RNA probe showed that the receptor gene is highly expressed in several hypothalamic and thalamic nuclei and to a lesser extent in the pallium and striatum. In the pituitary, the highest messenger RNA levels were detected in the distal lobe. Taken together, these data show that the cloned frog receptor shares several common features with both the VPAC1-R and VPAC2-R of mammals; the frog receptor exhibits the highest sequence identity with mammalian VPAC1-R, but the lack of effect of secretin and the brain distribution of the receptor are reminiscent of the characteristics of the mammalian VPAC2-R. The sequence of the frog receptor should thus prove useful to decipher the structure-activity relationships of the VIP/PACAP receptor family.


Association between sleep and Alzheimer's disease: A bibliometric analysis from 2003 to 2022.
Alzheimer's disease (AD) often presents with sleep disorders, which are also an important risk factor for AD, affecting cognitive function to a certain extent. This study aimed to reveal the current global status, present hotspots, and discuss emerging trends of sleep and AD using a bibliometric approach. Research and review articles related to sleep and AD from 2003 to 2022 were extracted from the Web of Science Core Collection. VOSviewer 220.127.116.11, Scimago Graphica, and CiteSpace 6.2.R2 were used to map the productive and highly cited countries, institutions, journals, authors, references, and keywords in the field. Overall, 4,008 publications were included in this bibliometric analysis. The number of publications and citations showed an increasing trend over the past two decades. The USA and China had the largest and second largest, respectively, number of publications and citations and cooperated with other countries more closely. Ancoli-Israel Sonia published the most papers, and Holtzman David M was co-cited most frequently. The most productive journal was Journal of Alzheimer's Disease, and Neurology was the most frequently cited journal. The risk factors, β-amyloid, tau, neuroinflammation, astrocytes, glymphatic system, orexin, functional connectivity, and management have been the main research directions of researchers over the past few years and may be the future trend of valuable research. We identified hotspots and emerging trends including risk factors, Aβ, tau, neuroinflammation, the glymphatic system, orexin, and management, which may help identify new therapeutic targets and improve clinical efficacy of sleep and AD.


Venous thromboembolism following 672,495 primary total shoulder and elbow replacements: Meta-analyses of incidence, temporal trends and potential risk factors.
There is wide variability in reported venous thromboembolism (VTE) incidence following total shoulder replacement (TSR) or total elbow replacement (TER). It is uncertain which risk factors influence the risk of VTE following TSR or TER. We conducted a PRISMA compliant meta-analysis to evaluate the incidence, temporal trends and potential risk factors for VTE following primary TSR and TER. MEDLINE, Embase, Web of Science, and Cochrane Library were searched to September 2019 for longitudinal studies reporting VTE outcomes after TSR or TER. Incidence and relative risks (RR) (95% confidence intervals) were estimated. We identified 43 articles with data on 672,495 TSRs and TERs (668,699 TSRs and 3796 TERs). The overall pooled 3-month VTE incidence following TSR was 0.85% (0.39-1.46). For TER, the 3-month incidence of VTE was 0.23% (0.08-0.44). Older age, body mass index (BMI) ≥25 kg/m2, and alcohol abuse were each associated with increased VTE risk following TSR. Comorbidities associated with increased VTE risk following TSR were chronic pulmonary disease, previous VTE, heart failure, anaemia, coagulopathy, arrhythmia, epilepsy, urinary tract infection, sleep apnoea, and fluid & electrolyte imbalance. Anatomic and outpatient TSR were each associated with decreased VTE risk. The average 3-month incidence of VTE following TSR or TER is <1%. High risk groups such as older patients, those with a previous VTE history and those undergoing reverse or inpatient TSR may need close monitoring. Modifiable factors such as high BMI, alcohol abuse, and comorbidities could be identified and addressed prior to surgery. PROSPERO 2019: CRD42019134096.


The quick inventory of depressive symptomatology (clinician and self-report versions) in patients with bipolar disorder.
The clinical and self-report versions of the Quick Inventory of Depressive Symptomatology (QIDS-C16 and QIDS-SR16) have been well studied in patients with major depressive disorder and in one recent study using patients with bipolar disorder. This article examines these measures in a second sample of 141 outpatients with bipolar disorder in different phases of the illness. At baseline, 61 patients were depressed and 30 were euthymic; at exit, 50 were depressed and 52 were euthymic. The remaining patients (at baseline or exit) were in either a manic or mixed phase and were pooled for statistical reasons. Similar results were found for the QIDS-C16 and QIDS-SR16. Scores were reasonably reliable to the extent that variability within groups permitted. As expected, euthymic patients showed less depressive symptomatology than depressed patients. Sad mood and general interest were the most discriminating symptoms between depressed and euthymic phases. Changes in illness phase (baseline to exit) were associated with substantial changes in scores. The relation of individual depressive symptoms to the overall level of depression was consistent across phases. Both the QIDS-SR16 and QIDS-C16 are suitable measures of depressive symptoms in patients with bipolar disorder.


[Optimal age of sexual maturity and limitations of juvenile survival in mammals].
Although the life history evolution of small- and large-bodied mammals seems to be governed by different factors, the both shows relative neonate size and juvenile survival to be slightly dependent on body mass. I propose a hypothesis that natural selection simultaneously maximizes a time to maturity (minimizes somatic growth rate) and a number of newborn survived to reproduction. In this case optimal juvenile survival of large-bodied mammals must be close to e-beta and that of small-bodied approximately e-(1 + beta), where beta is the slope of the regression of log annual fecundity on log annual juvenile mortality. Analysis of vital characters for 71 mammal species revealed the slope to be close to unity. As a result frequency distribution of log juvenile survival shows bimodality which coincides well with predicted optimal survival for large- and small-bodies species. It is shown that the relative neonate size can be directly proportional to the juvenile survival and inversely proportional to the lifetime offspring production irrespective of mortality factors.


Land-atmospheric feedbacks during droughts and heatwaves: state of the science and current challenges.
Droughts and heatwaves cause agricultural loss, forest mortality, and drinking water scarcity, especially when they occur simultaneously as combined events. Their predicted increase in recurrence and intensity poses serious threats to future food security. Still today, the knowledge of how droughts and heatwaves start and evolve remains limited, and so does our understanding of how climate change may affect them. Droughts and heatwaves have been suggested to intensify and propagate via land-atmosphere feedbacks. However, a global capacity to observe these processes is still lacking, and climate and forecast models are immature when it comes to representing the influences of land on temperature and rainfall. Key open questions remain in our goal to uncover the real importance of these feedbacks: What is the impact of the extreme meteorological conditions on ecosystem evaporation? How do these anomalies regulate the atmospheric boundary layer state (event self-intensification) and contribute to the inflow of heat and moisture to other regions (event self-propagation)? Can this knowledge on the role of land feedbacks, when available, be exploited to develop geo-engineering mitigation strategies that prevent these events from aggravating during their early stages? The goal of our perspective is not to present a convincing answer to these questions, but to assess the scientific progress to date, while highlighting new and innovative avenues to keep advancing our understanding in the future.


Xeroderma Pigmentosum Group C Deficiency Alters Cigarette Smoke DNA Damage Cell Fate and Accelerates Emphysema Development.
Cigarette smoke (CS) exposure is a major risk factor for the development of emphysema, a common disease characterized by loss of cells comprising the lung parenchyma. The mechanisms of cell injury leading to emphysema are not completely understood but are thought to involve persistent cytotoxic or mutagenic DNA damage induced by CS. Using complementary cell culture and mouse models of CS exposure, we investigated the role of the DNA repair protein, xeroderma pigmentosum group C (XPC), on CS-induced DNA damage repair and emphysema. Expression of XPC was decreased in mouse lungs after chronic CS exposure and XPC knockdown in cultured human lung epithelial cells decreased their survival after CS exposure due to activation of the intrinsic apoptosis pathway. Similarly, cell autophagy and apoptosis were increased in XPC-deficient mouse lungs and were further increased by CS exposure. XPC deficiency was associated with structural and functional changes characteristic of emphysema, which were worsened by age, similar to levels observed with chronic CS exposure. Taken together, these findings suggest that repair of DNA damage by XPC plays an important and previously unrecognized role in the maintenance of alveolar structures. These findings support that loss of XPC, possibly due to chronic CS exposure, promotes emphysema development and further supports a link between DNA damage, impaired DNA repair, and development of emphysema.


Effects of forest fragmentation on the seedling recruitment of a tropical herb: assessing seed vs. safe-site limitation.
Studies simultaneously evaluating the importance of safe-site and seed limitation for plant establishment are rare, particularly in human-modified landscapes. We used spatially explicit neighborhood models together with data from 10 0.5-ha mapped census plots in a fragmented landscape spanning 1000 km2 to (1) evaluate the relative importance of seed production, dispersal, and safe-site limitation for the recruitment of the understory herb Heliconia acuminata; and (2) determine how these processes differ between fragments and continuous forests. Our analyses demonstrated a large degree of variation in seed production, dispersal, and establishment among and within the 10 study plots. Seed production limitation was strong but only at small spatial scales. Average dispersal distance was less than 4 m, leading to severe dispersal limitation at most sites. Overall, safe-site limitation was the most important constraint on seedling establishment. Fragmentation led to a more heterogeneous light environment with negative consequences for seedling establishment but had little effect on seed production or dispersal. These results suggest that the effects of fragmentation on abiotic processes may be more important than the disruption of biotic interactions in driving biodiversity loss in tropical forests, at least for some functional groups. These effects may be common when the matrix surrounding fragments contains enough tree cover to enable movement of dispersers and pollinators.


Spectral characteristics of visible light-induced pigmentation and visible light protection factor.
Solar radiation is a major contributor to the development of skin cancer. Recent studies have shown that visible light (VL), a major portion of solar spectrum, induces biologic effects on the skin. Ultraviolet filters in currently available broad-spectrum sunscreens do not offer protection against VL. This study was designed to identify the spectral characteristics of the skin responses induced by VL, which can be utilized for time efficient in vivo VL testing. Thirty-one subjects were irradiated with a light source emitting visible light with less than 0.5% long wavelength UVA1 (VL + UVA1, 370-700 nm), and 41 subjects were irradiated with pure visible light (pure VL, 400-700 nm). Assessments including clinical photography, investigator's global assessment of pigmentation and erythema, and diffuse reflectance spectroscopy (DRS) performed immediately and seven days after irradiation. Clinical and spectroscopic data showed that VL + UVA1 spectral output induced significantly darker and persistent skin responses as compared to those induced by pure VL. Spectroscopic signatures of skin responses induced by both radiation sources were identified. The signatures were found to be specific to the radiation source and time of collection. A method to evaluate VL protection factor, using quantitative information from the spectral signatures obtained, was proposed.


Sam68 is required for the growth and survival of nonmelanoma skin cancer.
Although targeting DNA repair signaling pathways has emerged as a promising therapeutic for skin cancer, the relevance of DNA damage responses (DDR) in the development and survival of nonmelanoma skin cancer (NMSC), the most common type of skin cancer, remains obscure. Here, we report that Src-associated substrate during mitosis of 68 kDa (Sam68), an early signaling molecule in DDR, is elevated in skin tumor tissues derived from NMSC patients and skin lesions from Gli2-transgenic mice. Downregulation of Sam68 impacts the growth and survival of human tumor keratinocytes and genetic ablation of Sam68 delays the onset of basal cell carcinomas (BCC) in Gli2-transgenic mice. Moreover, Sam68 plays a critical role in DNA damage-induced DNA repair and nuclear factor kappa B (NF-κB) signaling pathways in keratinocytes, hence conferring keratinocyte sensitivity to DNA damaging agents. Together, our data reveal a novel function of Sam68 in regulating DDR in keratinocytes that is crucial for the growth and survival of NMSC.


Managing older adults with CKD: individualized versus disease-based approaches.
The last decade has seen the evolution and ongoing refinement of a disease-oriented approach to chronic kidney disease (CKD). Disease-oriented models of care assume a direct causal association between observed signs and symptoms and underlying disease pathophysiologic processes. Treatment plans target underlying disease mechanisms with the goal of improving disease-related outcomes. Because average glomerular filtrate rates tend to decrease with age, CKD becomes increasingly prevalent with advancing age and those who meet criteria for CKD are disproportionately elderly. However, several features of geriatric populations may limit the utility of disease-oriented models of care. In older adults, complex comorbid conditions and geriatric syndromes are common; signs and symptoms often do not reflect a single underlying pathophysiologic process; there can be substantial heterogeneity in life expectancy, functional status, and health priorities; and information about the safety and efficacy of recommended interventions often is lacking. For all these reasons, geriatricians have tended to favor an individualized patient-centered model of care over more traditional disease-based approaches. An individualized approach prioritizes patient preferences and embraces the notion that observed signs and symptoms often do not reflect a single unifying disease process and instead reflect the complex interplay between many different factors. This approach emphasizes modifiable outcomes that matter to the patient. Prognostic information related to these and other outcomes generally is used to shape rather than dictate treatment decisions. We argue that an individualized patient-centered approach to care may have more to offer than a traditional disease-based approach to CKD in many older adults.


Vib-PT, an Aromatic Prenyltransferase Involved in the Biosynthesis of Vibralactone from Stereum vibrans.
Vibralactone, a hybrid compound derived from phenols and a prenyl group, is a strong pancreatic lipase inhibitor with a rare fused bicyclic β-lactone skeleton. Recently, a researcher reported a vibralactone derivative (compound C1) that caused inhibition of pancreatic lipase with a half-maximal inhibitory concentration of 14 nM determined by structure-based optimization, suggesting a potential candidate as a new antiobesity treatment. In the present study, we sought to identify the main gene encoding prenyltransferase in Stereum vibrans, which is responsible for the prenylation of phenol leading to vibralactone synthesis. Two RNA silencing transformants of the identified gene (vib-PT) were obtained through Agrobacterium tumefaciens-mediated transformation. Compared to wild-type strains, the transformants showed a decrease in vib-PT expression ranging from 11.0 to 56.0% at 5, 10, and 15 days in reverse transcription-quantitative PCR analysis, along with a reduction in primary vibralactone production of 37 to 64% at 15 and 21 days, respectively, as determined using ultra-high-performance liquid chromatography-mass spectrometry analysis. A soluble and enzymatically active fusion Vib-PT protein was obtained by expressing vib-PT in Escherichia coli, and the enzyme's optimal reaction conditions and catalytic efficiency (Km /kcat) were determined. In vitro experiments established that Vib-PT catalyzed the C-prenylation at C-3 of 4-hydroxy-benzaldehyde and the O-prenylation at the 4-hydroxy of 4-hydroxy-benzenemethanol in the presence of dimethylallyl diphosphate. Moreover, Vib-PT shows promiscuity toward aromatic compounds and prenyl donors.IMPORTANCE Vibralactone is a lead compound with a novel skeleton structure that shows strong inhibitory activity against pancreatic lipase. Vibralactone is not encoded by the genome directly but rather is synthesized from phenol, followed by prenylation and other enzyme reactions. Here, we used an RNA silencing approach to identify and characterize a prenyltransferase in a basidiomycete species that is responsible for the synthesis of vibralactone. The identified gene, vib-PT, was expressed in Escherichia coli to obtain a soluble and enzymatically active fusion Vib-PT protein. In vitro characterization of the enzyme demonstrated the catalytic mechanism of prenylation and broad substrate range for different aromatic acceptors and prenyl donors. These characteristics highlight the possibility of Vib-PT to generate prenylated derivatives of aromatics and other compounds as improved bioactive agents or potential prodrugs.


Isolation of p53 protein from the serum of colon-cancer and noncancer patients.
A new method for the isolation of p53 protein from the sera of cancer and non-cancer patients has been developed. The method is based on the utilization of a new modification of a support for affinity chromatography in the form of gel fiberglass (GFG) (R. Zusman, patent applications, Israel, 1992; USA 1993). The sera were percolated through GFG columns with entrapped rabbit IgG generated against the colon cancer antigens. Tumor-associated antigens were eluted from sera of cancer patients in large amounts, up to 2.4 mg/ml serum/column. Concentrations of these antigens in different sera and their affinity to anti-tumoral antibodies were detected by ELISA. Western immunoblotting with commercial monoclonal antibodies and SDS-PAGE identified the isolated antigens as the p53 protein. This protein has been isolated in the highest concentration yet reported from the serum of cancer patients, up to 1 mg/ml. The same protein has been found in the serum of healthy people and non-cancer patients but in significantly lower concentrations. Isolation of p53 protein from sera of noncancer patients confirms the opinion of some investigators that this protein should be considered not only as an oncogene but also as a protein correlated with different changes in the cell cycle.


To center or not to center? Investigating inertia with a multilevel autoregressive model.
Whether level 1 predictors should be centered per cluster has received considerable attention in the multilevel literature. While most agree that there is no one preferred approach, it has also been argued that cluster mean centering is desirable when the within-cluster slope and the between-cluster slope are expected to deviate, and the main interest is in the within-cluster slope. However, we show in a series of simulations that if one has a multilevel autoregressive model in which the level 1 predictor is the lagged outcome variable (i.e., the outcome variable at the previous occasion), cluster mean centering will in general lead to a downward bias in the parameter estimate of the within-cluster slope (i.e., the autoregressive relationship). This is particularly relevant if the main question is whether there is on average an autoregressive effect. Nonetheless, we show that if the main interest is in estimating the effect of a level 2 predictor on the autoregressive parameter (i.e., a cross-level interaction), cluster mean centering should be preferred over other forms of centering. Hence, researchers should be clear on what is considered the main goal of their study, and base their choice of centering method on this when using a multilevel autoregressive model.


Chronic Cannabidiol Administration Fails to Diminish Blood Pressure in Rats with Primary and Secondary Hypertension Despite Its Effects on Cardiac and Plasma Endocannabinoid System, Oxidative Stress and Lipid Metabolism.
We investigated the influence of cannabidiol (CBD) on blood pressure (BP) and heart rate (HR) in spontaneously (SHR) and deoxycorticosterone (DOCA-salt) hypertensive rats. Hypertension was connected with increases in cardiac and plasma markers of lipid peroxidation in both models, whereas cardiac endocannabinoid levels decreased in SHR and increased in DOCA-salt. CBD (10 mg/kg once a day for 2 weeks) did not modify BP and HR in hypertension but counteracted pro-oxidant effects. Moreover, it decreased cardiac or plasma levels of anandamide, 2-arachidonoylglycerol and oleoyl ethanolamide in DOCA-salt and inhibited the activity of fatty acid amide hydrolase (FAAH) in both models. In the respective normotensive control rats, CBD increased lipid peroxidation, free fatty acid levels and FAAH activity. In conclusion, chronic CBD administration does not possess antihypertensive activity in a model of primary and secondary (DOCA-salt) hypertension, despite its antioxidant effect. The latter may be direct rather than based on the endocannabinoid system. The unexpected CBD-related increase in lipid peroxidation in normotensive controls may lead to untoward effects; thus, caution should be kept if CBD is used therapeutically.


Wheat hybridization and polyploidization results in deregulation of small RNAs.
Speciation via interspecific or intergeneric hybridization and polyploidization triggers genomic responses involving genetic and epigenetic alterations. Such modifications may be induced by small RNAs, which affect key cellular processes, including gene expression, chromatin structure, cytosine methylation and transposable element (TE) activity. To date, the role of small RNAs in the context of wide hybridization and polyploidization has received little attention. In this work, we performed high-throughput sequencing of small RNAs of parental, intergeneric hybrid, and allopolyploid plants that mimic the genomic changes occurring during bread wheat speciation. We found that the percentage of small RNAs corresponding to miRNAs increased with ploidy level, while the percentage of siRNAs corresponding to TEs decreased. The abundance of most miRNA species was similar to midparent values in the hybrid, with some deviations, as seen in overrepresentation of miR168, in the allopolyploid. In contrast, the number of siRNAs corresponding to TEs strongly decreased upon allopolyploidization, but not upon hybridization. The reduction in corresponding siRNAs, together with decreased CpG methylation, as shown here for the Veju element, represent hallmarks of TE activation. TE-siRNA downregulation in the allopolyploid may contribute to genome destabilization at the initial stages of speciation. This phenomenon is reminiscent of hybrid dysgenesis in Drosophila.


Altered ganglioside expression modulates the pathogenic mechanism of thyroid-associated ophthalmopathy by increase in hyaluronic acid.
The aim of this study was to determine the role of gangliosides in pathogenic mechanisms of thyroid-associated ophthalmopathy (TAO). The gangliosides profile and mRNA level of sialyltransferases of the orbital tissues from TAO patients (n = 5) and non-TAO subjects (n = 4) were investigated. In addition, the effect of exogenous gangliosides on the expression of hyaluronic acid was examined in orbital fibroblasts. For in vitro experiments, we used four different strains of cells obtained from non-TAO subjects with at least three replicates for each strain. Trisialoganglioside 1b (GT1b) was significantly overexpressed in the orbital tissue of TAO patients compared with control tissue, whereas no significant difference was observed for either monosialoganglioside 1 (GM1) or disialoganglioside 1a (GD1a) by digital analyses of immunohistochemical images. Moreover, mRNA levels of sialyltransferase (SAT)-I and SAT II were increased in TAO patients compared with control. Exogenous GT1b strongly induced the morphologic changes related to an accumulation of sparse flocculent precipitates in lysosomes and increased the extracellular hyaluronic acid level in orbital fibroblasts with the induction of hyaluronic acid synthase, which were less by GD1a but not by GM1. The GT1b-induced morphologic changes of cells were due, at least in part, to an increase of intracellular hyaluronic acid. Co-treatment of hyaluronidase nicely attenuated the morphologic changes in orbital fibroblasts. Thy-1(+) orbital fibroblasts were more capable of producing hyaluronic acid by exogenous GT1b. The results suggest that gangliosides, particularly GT1b, may play a role in the pathologic mechanisms of TAO by stimulating an increase in hyaluronic acid.


The surgical management of prolapsed pedunculated submucous leiomyomas.
Prolapsed pedunculated leiomyomas of the uterus can best be managed by simple vaginal myomectomy. It is safe, easily performed and generally requires no anesthesia. The risk of complications during and after a major abdominal surgical procedure in the face of infection and anemia is eliminated. Interval hysterectomy, if indicated, may be done four to six weeks after vaginal myomectomy without incurring the additional risk of increased operative morbidity. Additional surgical procedures are not indicated if the pelvic examination remains normal. When large leiomyomas are encountered, the pedicle may not be accessible. Confronted with such a situation and continued blood loss and sepsis, we elected in three patients to replace the tumor within the endometrial cavity and close the cervix. Immediate hysterectomy was then carried out. Blood replacementd antibiotic coverage were instituted in advance of any operative procedure. Other pathologic entities must be kept in mind as a source of uterine bleeding that may be found in association with pedunculated myomas.


Phage Therapy in the Postantibiotic Era.
Antibiotic resistance is arguably the biggest current threat to global health. An increasing number of infections are becoming harder or almost impossible to treat, carrying high morbidity, mortality, and financial cost. The therapeutic use of bacteriophages, viruses that infect and kill bacteria, is well suited to be part of the multidimensional strategies to combat antibiotic resistance. Although phage therapy was first implemented almost a century ago, it was brought to a standstill after the successful introduction of antibiotics. Now, with the rise of antibiotic resistance, phage therapy is experiencing a well-deserved rebirth. Among the admittedly vast literature recently published on this topic, this review aims to provide a forward-looking perspective on phage therapy and its role in modern society. We cover the key points of the antibiotic resistance crisis and then explain the biological and evolutionary principles that support the use of phages, their interaction with the immune system, and a comparison with antibiotic therapy. By going through up-to-date reports and, whenever possible, human clinical trials, we examine the versatility of phage therapy. We discuss conventional approaches as well as novel strategies, including the use of phage-antibiotic combinations, phage-derived enzymes, exploitation of phage resistance mechanisms, and phage bioengineering. Finally, we discuss the benefits of phage therapy beyond the clinical perspective, including opportunities for scientific outreach and effective education, interdisciplinary collaboration, cultural and economic growth, and even innovative use of social media, making the case that phage therapy is more than just an alternative to antibiotics.


Initial characterization of the FlgE hook high molecular weight complex of Borrelia burgdorferi.
The spirochete periplasmic flagellum has many unique attributes. One unusual characteristic is the flagellar hook. This structure serves as a universal joint coupling rotation of the membrane-bound motor to the flagellar filament. The hook is comprised of about 120 FlgE monomers, and in most bacteria these structures readily dissociate to monomers (∼ 50 kDa) when treated with heat and detergent. However, in spirochetes the FlgE monomers form a large mass of over 250 kDa [referred to as a high molecular weight complex (HMWC)] that is stable to these and other denaturing conditions. In this communication, we examined specific aspects with respect to the formation and structure of this complex. We found that the Lyme disease spirochete Borrelia burgdorferi synthesized the HMWC throughout the in vitro growth cycle, and also in vivo when implanted in dialysis membrane chambers in rats. The HMWC was stable to formic acid, which supports the concept that the stability of the HMWC is dependent on covalent cross-linking of individual FlgE subunits. Mass spectrometry analysis of the HMWC from both wild type periplasmic flagella and polyhooks from a newly constructed ΔfliK mutant indicated that other proteins besides FlgE were not covalently joined to the complex, and that FlgE was the sole component of the complex. In addition, mass spectrometry analysis also indicated that the HMWC was composed of a polymer of the FlgE protein with both the N- and C-terminal regions remaining intact. These initial studies set the stage for a detailed characterization of the HMWC. Covalent cross-linking of FlgE with the accompanying formation of the HMWC we propose strengthens the hook structure for optimal spirochete motility.


The origin of multiply sigmoid curves of pH-dependence. The partitioning of groups among titration pK values.
An acid, HnA, with n ionizing groups is known to have the same titration curve as an equimolar mixture of n hypothetical monobasic acids, whose dissociation constants are known as the 'titration constants' of the real acid. We show that the pH-dependence of any property of HnA is also represented by the sum of one-site titration curves, characterized by these same titration constants. Since one such property is the degree of dissociation of one of the dissociating groups, a fraction of each group shows each of the various titration pK values, so that the group partitions among them. The n groups therefore share the same n titration pK values but differ in the fractions belonging to each. The one H+ ion per molecule that titrates with each pK is thus made up of the fractions, one from each group, that share this pK value. A group may possess a single pK value, in that it contributes virtually all of this pK and almost nothing to the others, only if either (1) in titrates in a different pH range from the other groups or (2) its affinity for H+ is unaffected by their ionization state.


Ancient AMY1 gene duplications primed the amylase locus for adaptive evolution upon the onset of agriculture.
Starch digestion is a cornerstone of human nutrition. The amylase enzyme, which digests starch, plays a key role in starch metabolism. Indeed, the copy number of the human amylase gene has been associated with metabolic diseases and adaptation to agricultural diets. Previous studies suggested that duplications of the salivary amylase gene are of recent origin. In the course of characterizing 51 distinct amylase haplotypes across 98 individuals employing long-read DNA sequencing and optical mapping methods, we detected four 31mers linked to duplication of the amylase locus. Analyses with these 31mers suggest that the first duplication of the amylase locus occurred more than 700,000 years ago before the split between modern humans and Neanderthals. After the original duplication events, amplification of the AMY1 genes likely occurred via nonallelic homologous recombination in a manner that consistently results in an odd number of copies per chromosome. These findings suggest that amylase haplotypes may have been primed for bursts of natural-selection associated duplications that coincided with the incorporation of starch into human diets.


Deaths: final data for 1998.
This report presents final 1998 data on U.S. deaths and death rates according to demographic and medical characteristics such as age, sex, race, Hispanic origin, marital status, educational attainment, injury at work, State of residence, and cause of death. Trends and patterns in general mortality, life expectancy, and infant and maternal mortality are also described. A previous report presented preliminary mortality data for 1998. In 1998 a total of 2,337,256 deaths were reported in the United States. This report presents descriptive tabulations of information reported on the death certificates. Death certificates are completed by funeral directors, attending physicians, medical examiners, and coroners. Original records are filed in the State registration offices. Statistical information is compiled into a national data base through the Vital Statistics Cooperative Program of the National Center for Health Statistics (NCHS), Centers for Disease Control and Prevention. The 1998 age-adjusted death rate for the United States decreased to an all-time low of 471.7 deaths per 100,000 standard population, and life expectancy at birth increased to a record high of 76.7 years. Of the 15 leading causes of death in 1998, the largest decline from the previous year--9.5 percent--in age-adjusted death rates was for Atherosclerosis (atherosclerosis). Human immunodeficiency virus (HIV) infection dropped from among the 15 leading causes for the first time since 1987. The age-adjusted death rate for firearm injuries decreased for the fifth consecutive year, declining 7.4 percent between 1997 and 1998. Among all causes of death, age-specific death rates rose for those under 1 year but declined for all other age groups, although the decline for children aged 1-4 years was not significant. The infant mortality rate was unchanged from 1997 at 7.2 infant deaths per 1,000 live births. The overall improvements in general mortality and life expectancy in 1998 continue the long-term downward trend in U.S. mortality. Although unchanged from 1997, the trend in U.S. infant mortality is of steady declines over the past four decades.


A mitochondrial superoxide signal triggers increased longevity in Caenorhabditis elegans.
The nuo-6 and isp-1 genes of C. elegans encode, respectively, subunits of complex I and III of the mitochondrial respiratory chain. Partial loss-of-function mutations in these genes decrease electron transport and greatly increase the longevity of C. elegans by a mechanism that is distinct from that induced by reducing their level of expression by RNAi. Electron transport is a major source of the superoxide anion (O(⋅) (-)), which in turn generates several types of toxic reactive oxygen species (ROS), and aging is accompanied by increased oxidative stress, which is an imbalance between the generation and detoxification of ROS. These observations have suggested that the longevity of such mitochondrial mutants might result from a reduction in ROS generation, which would be consistent with the mitochondrial oxidative stress theory of aging. It is difficult to measure ROS directly in living animals, and this has held back progress in determining their function in aging. Here we have adapted a technique of flow cytometry to directly measure ROS levels in isolated mitochondria to show that the generation of superoxide is elevated in the nuo-6 and isp-1 mitochondrial mutants, although overall ROS levels are not, and oxidative stress is low. Furthermore, we show that this elevation is necessary and sufficient to increase longevity, as it is abolished by the antioxidants NAC and vitamin C, and phenocopied by mild treatment with the prooxidant paraquat. Furthermore, the absence of effect of NAC and the additivity of the effect of paraquat on a variety of long- and short-lived mutants suggest that the pathway triggered by mitochondrial superoxide is distinct from previously studied mechanisms, including insulin signaling, dietary restriction, ubiquinone deficiency, the hypoxic response, and hormesis. These findings are not consistent with the mitochondrial oxidative stress theory of aging. Instead they show that increased superoxide generation acts as a signal in young mutant animals to trigger changes of gene expression that prevent or attenuate the effects of subsequent aging. We propose that superoxide is generated as a protective signal in response to molecular damage sustained during wild-type aging as well. This model provides a new explanation for the well-documented correlation between ROS and the aged phenotype as a gradual increase of molecular damage during aging would trigger a gradually stronger ROS response.


Activation and inhibition of benzo(a)pyrene and aflatoxin B1 metabolism in human liver microsomes by naturally occurring flavonoids.
The effects of several naturally occurring and synthetic flavonoids on the metabolism of benzo(a)pyrene and aflatoxin B1 were evaluated. Addition of apigenin, chrysin, fisetin, flavonone, galangin, hesperitin, kaempferol, morin, myricetin, haringenin, or quercetin to human liver microsomes inhibited the hydroxylation of benzo(a)pyrene. In contrast to these results, the addition of flavone, nobiletin, tangeretin, or 7,8-benzoflavone to human liver microsomes caused a many-fold stimulation in the hydroxylation of benzo(a)pyrene, the metabolism of aflatoxin B1 to 2,3-dihydro-2,3-dihydroxyaflatoxin B1, and the metabolic activation of aflatoxin B1 to mutagenic products. Quercetin, morin, and kaempferol inhibited cytochrome c (P-450) reductase in human liver microsomes whereas flavone and 7,8-benzoflavone had no effect. These results suggest that the inhibitory effects of quercetin, morin, and kaempferol on monooxygenase activity may be caused at least in part by an inhibition in the reduction of cytochrome P-450. An examination of the structural features required for the inhibition and stimulation of benzo(a)pyrene hydroxylation indicated that all of the 12 flavonoid inhibitors that were studied possessed hydroxyl groups whereas the flavonoid activators were less polar molecules that lacked hydroxyl groups.


Light and electron microscopic localization of D-galactosyl residues in capillary endothelial cells of the canine cerebral cortex.
Ricinus communis agglutinin I (RCA-I), a lectin that binds to D-galactosyl residues, intensely stained capillaries in cryostat sections of canine cerebral cortex when evaluated by the avidin-biotin-peroxidase complex method. Of seven lectins tested, only RCA-I gave strong staining of vessels and capillaries with little staining of other cortical cells. Ultrastructural studies using ferritin-, biotin-, and peroxidase-labeled RCA-I indicated that this lectin was bound to the luminal membrane of the cerebral capillary endothelial cell and that lectin receptors were distributed continuously along this membrane. Plasmalemma invaginations that bound RCA-I were also present in endothelial cells. Primary cultures of cerebral capillary endothelial cells grown on plastic or gelatin-coated glass substrates demonstrated staining of the cell membrane and perinuclear structures which appeared to be the Golgi complex and secondary lysosomes. These staining characteristics were retained when the cells were subcultured and were confirmed by ultrastructural studies. In contrast, light microscopy showed that fibronectin was more widely distributed in the cytoplasm, a finding consistent with its occurrence in the endoplasmic reticulum. This work provides support for the concept that lectins may be useful endothelial cell markers in both intact tissue and cell culture.


Microcirculation in Hypertension: An Update on Clinical Significance and Therapy.
Essential hypertension is closely related to alterations of the microcirculation as evidenced by increased media-to-lumen ratio of small resistance arteries and capillary rarefaction. It has been proposed that microvasculature alterations can further deteriorate target organ damage under the influence of increased hemodynamic load in hypertension. More than a decade ago, small artery structure was found to be the most potent predictor of cardiovascular events in essential or secondary hypertension. Amelioration of resistance vessel structure by antihypertensive drugs showed a clear benefit when patients were treated with calcium antagonists and renin-angiotensin-aldosterone inhibitor regimens. Well-designed controlled studies are needed to address the important questions whether different antihypertensive agent combinations could be related to diverse microcirculatory "responses" in terms of structure and function.


$L_0$ Regularized Stationary-Time Estimation for Crowd Analysis.
In this paper, we tackle the problem of stationary crowd analysis which is as important as modeling mobile groups in crowd scenes and finds many important applications in crowd surveillance. Our key contribution is to propose a robust algorithm for estimating how long a foreground pixel becomes stationary. It is much more challenging than only subtracting background because failure at a single frame due to local movement of objects, lighting variation, and occlusion could lead to large errors on stationary-time estimation. To achieve robust and accurate estimation, sparse constraints along spatial and temporal dimensions are jointly added by mixed partials (which are second-order gradients) to shape a 3D stationary-time map. It is formulated as an L0 optimization problem. Besides background subtraction, it distinguishes among different foreground objects, which are close or overlapped in the spatio-temporal space by using a locally shared foreground codebook. The proposed technologies are further demonstrated through three applications. 1) Based on the results of stationary-time estimation, 12 descriptors are proposed to detect four types of stationary crowd activities. 2) The averaged stationary-time map is estimated to analyze crowd scene structures. 3) The result of stationary-time estimation is also used to study the influence of stationary crowd groups to traffic patterns.