Elucidating Proton-Coupled Electron Transfer Mechanisms of Metal Hydrides with Free Energy- and Pressure-Dependent Kinetics.
Proton-coupled electron transfer (PCET) was studied in a series of tungsten hydride complexes with pendant pyridyl arms ([(PyCH2Cp)WH(CO)3], PyCH2Cp = pyridylmethylcyclopentadienyl), triggered by laser flash-generated RuIII-tris-bipyridine oxidants, in acetonitrile solution. The free energy dependence of the rate constant and the kinetic isotope effects (KIEs) showed that the PCET mechanism could be switched between concerted and the two stepwise PCET mechanisms (electron-first or proton-first) in a predictable fashion. Straightforward and general guidelines for how the relative rates of the different mechanisms depend on oxidant and base are presented. The rate of the concerted reaction should depend symmetrically on changes in oxidant and base strength, that is on the overall ΔG0PCET, and we argue that an "asynchronous" behavior would not be consistent with a model where the electron and proton tunnel from a common transition state. The observed rate constants and KIEs were examined as a function of hydrostatic pressure (1-2000 bar) and were found to exhibit qualitatively different dependence on pressure for different PCET mechanisms. This is discussed in terms of different volume profiles of the PCET mechanisms as well as enhanced proton tunneling for the concerted mechanism. The results allowed for assignment of the main mechanism operating in the different cases, which is one of the critical questions in PCET research. They also show how the rate of a PCET reaction will be affected very differently by changes of oxidant and base strength, depending on which mechanism dominates. This is of fundamental interest as well as of practical importance for rational design of, for example, catalysts for fuel cells and solar fuel formation, which operate in steps of PCET reactions. The mechanistic richness shown by this system illustrates that the specific mechanism is not intrinsic to a specific synthetic catalyst or enzyme active site but depends on the reaction conditions.


Septic impairment of capillary blood flow requires nicotinamide adenine dinucleotide phosphate oxidase but not nitric oxide synthase and is rapidly reversed by ascorbate through an endothelial nitric oxide synthase-dependent mechanism.
To determine the roles of nitric oxide synthase (NOS) and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in the impairment of capillary blood flow in sepsis and in the reversal of this impairment by ascorbate. Prospective, controlled laboratory study. Animal laboratory in research institute. Adult male wild type (WT), neuronal nitric oxide synthase (nNOS)-/-, inducible NOS (iNOS)-/-, endothelial NOS (eNOS)-/-, and gp91phox-/- mice. Sepsis was induced by feces injection into peritoneum (FIP). A bolus of ascorbate or NADPH oxidase inhibitor apocynin was injected intravenously at 6 hrs post-FIP. Alternatively, NOS cofactor (6R)-5,6,7,8-tetrahydro-L-biopterin (BH4) or nitric oxide donor S-nitroso-N-acetylpenicillamine was superfused on the surface of the extensor digitorum longus muscle. Capillary blood flow impairment and NOS activity in the extensor digitorum longus muscle were measured by intravital microscopy and by enzymatic assay, respectively. Sepsis at 6 hrs impaired flow in WT mice. Apocynin, and knockout of gp91phox but not of any NOS isoforms, rescued this impairment. Constitutive NOS activity was unaffected by sepsis, but it was abolished by nNOS knockout (iNOS activity was negligible in all mice). Ascorbate rapidly (10 mins) rescued impaired flow in WT, nNOS-/-, iNOS-/- but not eNOS-/- mice. Ascorbate also improved survival of WT mice after FIP. BH4 and SNAP rescued flow in WT mice, while BH4 failed to rescue it in eNOS-/- mice. Capillary blood flow impairment in septic skeletal muscle requires NADPH oxidase but not NOS, and it is rapidly reversed by ascorbate and BH4 through an eNOS-dependent mechanism.


The development of diabetes mellitus and chronic liver disease in long term chelated beta thalassaemic patients.
We studied 29 patients with thalassaemia major who had received intensive chelation for between 6.2 and 8.8 years. All patients had normal oral glucose tolerance tests before subcutaneous chelation therapy was introduced and 22 of 29 patients had normal liver function tests. At the end of the period of study 12 patients still had normal oral glucose tolerance (7 with normal liver function tests and 5 with chronic active hepatitis). On the other hand, 11 patients had developed impaired glucose tolerance tests (3 patients had normal liver function tests, 5 with chronic active hepatitis and 3 with cirrhosis), and 6 patients had developed frank diabetes mellitus (one with chronic active hepatitis and 5 with cirrhosis). Patients with chronic active hepatitis showed 91% positivity for one or more hepatitis B markers whilst all patients with cirrhosis were positive. Ferritin levels before subcutaneous chelation in patients with normal oral glucose tolerance tests were lower than in those patients with abnormal oral glucose tolerance or diabetes (P less than 0.05) but none had normal serum ferritin levels. In addition, a positive correlation was found between glucose area under the curve after chelation therapy and serum ferritin levels (r = 0.47, P less than 0.01). It is apparent that long term chelation therapy does not prevent the development of abnormal oral glucose tolerance in chronically transfused patients. More intensive chelation therapy is needed to prevent tissue damage. Chronic liver disease may have an important role to play in the deterioration of glucose tolerance.


Costs of Malnutrition in Institutionalized and Community-Dwelling Older Adults: A Systematic Review.
The aim of this study was to assess health economics evidence published to date on malnutrition costs in institutionalized or community-dwelling older adults. A systematic search of the literature published until December 2013 was performed using standard literature, international and national electronic databases, including MedLine/PubMed, Cochrane Library, ISI WOK, SCOPUS, MEDES, IBECS, and Google Scholar. Publications identified referred to the economic burden and use of medical resources associated with malnutrition (or risk of malnutrition) in institutionalized or community-dwelling older adults, written in either English or Spanish. Costs were updated to 2014 (€). A total of 9 studies of 46 initially retrieved met the preestablished criteria and were submitted to thorough scrutiny. All publications reviewed involved studies conducted in Europe, and the results regarding the contents of all the studies showed that total costs associated with malnutrition in institutionalized and community-dwelling older adults were considerably higher than those of well-nourished ones, mainly due to a higher use of health care resources (GP consultations, hospitalizations, health care monitoring, and treatments). Interventions to reduce the prevalence of malnutrition, such as the use of oral nutritional supplements, showed an important decrease in-hospital admissions and medical visits. Malnutrition is associated with higher health care costs in institutionalized or community-dwelling older adults. The adoption of nutritional interventions, such as oral nutritional supplements, may have an important impact in reducing annual health care costs per patient.


The role of the psychiatrist in evaluating a prison mental health system in litigation.
Federal courts during the past 14 years have recognized that many prisons in the United States have provided constitutionally inadequate medical and psychiatric services. Our recent national survey indicates that at least 20 states have had at least one part of their correctional system included in a certified class action suit that alleged insufficient mental health services for inmates. This article reviews the role of the expert psychiatric witness during the phases of litigation that involve proposed remedial plans and compliance in implementing remedial plans. Available epidemiologic data about psychiatric disorders among prison inmates, standards for correctional mental health care, and various mental health system models are briefly reviewed. A comprehensive approach to evaluating proposed remedial plans and assessing issues of compliance with accepted plans is described. Special attention is directed toward psychiatric issues unique to a correctional system.


An Investigation of Atomic Structures Derived from X-ray Crystallography and Cryo-Electron Microscopy Using Distal Blocks of Side-Chains.
Cryo-electron microscopy (cryo-EM) is a structure determination method for large molecular complexes. As more and more atomic structures are determined using this technique, it is becoming possible to perform statistical characterization of side-chain conformations. Two data sets were involved to characterize block lengths for each of the 18 types of amino acids. One set contains 9131 structures resolved using X-ray crystallography from density maps with better than or equal to 1.5 Å resolutions, and the other contains 237 protein structures derived from cryo-EM density maps with 2-4 Å resolutions. The results show that the normalized probability density function of block lengths is similar between the X-ray data set and the cryo-EM data set for most of the residue types, but differences were observed for ARG, GLU, ILE, LYS, PHE, TRP, and TYR for which conformations with certain shorter block lengths are more likely to be observed in the cryo-EM set with 2-4 Å resolutions.


Scalable transfer of vertical graphene nanosheets for flexible supercapacitor applications.
Vertical graphene nanosheets (VGN) are the material of choice for application in next-generation electronic devices. The growing demand for VGN-based flexible devices for the electronics industry brings in restriction on VGN growth temperature. The difficulty associated with the direct growth of VGN on flexible substrates can be overcome by adopting an effective strategy of transferring the well-grown VGN onto arbitrary flexible substrates through a soft chemistry route. In the present study, we report an inexpensive and scalable technique for the polymer-free transfer of VGN onto arbitrary substrates without disrupting its morphology, structure, and properties. After transfer, the morphology, chemical structure, and electrical properties are analyzed by scanning electron microscopy, Raman spectroscopy, x-ray photoelectron spectroscopy, and four-probe resistive methods, respectively. The wetting properties are studied from the water contact angle measurements. The observed results indicate the retention of morphology, surface chemistry, structure, and electronic properties. Furthermore, the storage capacity of the transferred VGN-based binder-free and current collector-free flexible symmetric supercapacitor device is studied. A very low sheet resistance of 670 Ω/□ and excellent supercapacitance of 158 μF cm-2 with 86% retention after 10 000 cycles show the prospect of the damage-free VGN transfer approach for the fabrication of flexible nanoelectronic devices.


Electrochemical preparation of zinc oxide/polypyrrole nanocomposite coating for the highly effective solid-phase microextraction of phthalate esters.
In this work, zinc oxide/polypyrrole nanocomposite coating was fabricated on stainless steel and evaluated as a novel headspace solid-phase microextraction fiber coating for extraction of ultra-trace amounts of environmental pollutants, namely, phthalate esters, in water samples. The fiber nanocomposite were prepared by a two-step process including the electrochemical deposition of polypyrrole on the surface of stainless steel in the first step, and electrochemical deposition of zinc oxide nanosheets in the second step. Porous structure together with zinc oxide nanosheets with the average diameter of 30 nm were observed on the surface by using scanning electron microscopy. The effective parameters on extraction of phthalate esters (i.e., extraction temperature, extraction time, desorption temperature, desorption time, salt concentration, and stirring rate) were investigated and optimized by one-variable-at-a-time method. Under optimized conditions (extraction temperature, 90°C; extraction time, 40 min; desorption temperature, 270°C; desorption time, 5 min; salt concentration, 25% w/v; and stirring rate, 1000 rpm), the limits of detection were in the range of 0.05-0.8 μg/L, and the repeatability and fiber-to-fiber reproducibility were in the ranges of 6.1-7.3% and 8.7-10.2%, respectively.


Plaque Development and Induction of Interferon Synthesis by RMC Poliovirus.
Johnson, Terry C. (University of Minnesota, Minneapolis), and Leroy C. McLaren. Plaque development and induction of interferon synthesis by RMC poliovirus. J. Bacteriol. 90:565-570. 1965.-Plaque development by RMC poliovirus on human amnion cell monolayers was investigated with regard to autointerference and to the effect of acid-agar overlay on plaquing efficiency. The virus was inhibited by acid-agar overlay, thereby exhibiting the d(-) marker typical of attenuated poliovirus strains. In addition, a lack of RMC poliovirus plaque development on HeLa cell monolayers was shown to be the result of an agar inhibitor which could be removed by NaCl extraction. By use of a simplified plaque reduction assay, it was shown that interferon production was responsible for the autointerference phenomenon. Interferon synthesis did not correlate with the ages in vitro of human amnion cell cultures. Fibroblasts originating from the chorionic membrane produced negligible amounts of the inhibitor. Interferon synthesis by human amnion cells infected with RMC poliovirus was inhibited by actinomycin D. The addition of guanidine hydrochloride to infected cultures immediately after RMC poliovirus adsorption markedly inhibited interferon synthesis, although after 2 hr (postadsorption) guanidine had no effect on interferon production.


Low concentrations of vinflunine induce apoptosis in human SK-N-SH neuroblastoma cells through a postmitotic G1 arrest and a mitochondrial pathway.
Vinflunine, the newest fluorinated Vinca alkaloid, currently in phase III clinical trials, targets the microtubule network to induce mitotic block and apoptosis by mechanisms that remain unclear. In the current study, we investigated the apoptotic pathways induced by a wide range of vinflunine concentrations in SK-N-SH neuroblastoma cells. The concentrations of vinflunine that inhibited 50 and 70% of cell growth (IC(50) and IC(70)) induced high extents of apoptosis but failed to depolymerize microtubule network and to block cells in G(2)/M. It is interesting that the IC(50) and IC(70) concentrations suppressed microtubule dynamics, slowed down mitotic progression from metaphase to anaphase, and induced a postmitotic G(1) arrest. This G(1) arrest was associated with an increase in p53 and p21 expression and with their nuclear translocation. A high concentration of vinflunine (500 nM) induced both microtubule depolymerization and a canonical G(2)/M block. Mitochondria were involved in apoptotic pathways because all studied concentrations induced cytochrome c release. Bcl-2 family members were differently modulated by the different drug concentrations. Bax was up-regulated and translocated to mitochondria at the IC(50) and IC(70) concentrations, whereas Bcl-2 was phosphorylated only at the highest vinflunine concentration examined (500 nM). Our findings can be extended to other Vinca alkaloids, because similar results were obtained with vinblastine. All together, our results show that low concentrations of vinflunine fail to promote a G(2)/M arrest but are sufficient to induce suppression of microtubule dynamics and subsequent apoptosis. Moreover, mitochondria constitute the point of convergence of apoptotic signals induced by both low and high concentrations of vinflunine.


Metacarpophalangeal pattern profile analysis in Noonan syndrome.
Metacarpophalangeal pattern (MCPP) analysis is an application of an anthropometric technique that provides a quantitative assessment of the amount and direction of abnormality in the hand skeleton. MCPP analysis was undertaken on 15 individuals (9 males, 6 females) with Noonan syndrome ranging in age from 0.1 to 36 years with a mean age at 11.6 years. The overall average Z score for the MCPP variables was -2.1 and the range was -2.5 (for metacarpal two) and -1.5 (for middle phalanx 5). The average hand pattern variability index, a measure of hand bone length relationships, was abnormal. A Pearsonian correlation analysis was used to assess similarity between the mean pattern and each of the 15 individual patterns. Nine (60%) of the fifteen individuals with Noonan syndrome had significant positive correlations (P < 0.05), indicating homogeneity or similarity in the hand patterns. A stepwise discriminant analysis was performed on Z score data from the individual hand bone measurements on the 15 subjects with Noonan syndrome and 41 healthy controls (24 females, 17 males; mean age = 13.1 years with age range of 9.6 to 18 years). This analysis produced a discriminant function with two MCPP variables (metacarpal 1 and middle phalanx 3) entering into the function and producing a correct classification rate of 93%. The two MCPP variables contributed to the overall difference between individuals with Noonan syndrome and the normative sample. The hand pattern variability index was outside of the normal range, indicating an abnormal MCPP with multivariate analysis. The MCPP analysis may be useful as a tool for diagnosis in screening subjects for Noonan syndrome.


[Less fat or a different fat?].
Current dietary advice with a view to avoiding cardiovascular pathology is to replace fatty foodstuffs and those rich in saturated fat and cholesterol by food rich in complex carbohydrates. Although substitution of carbohydrates for fat lowers the blood level of low-density lipoprotein (LDL) cholesterol, it also lowers the level of high-density lipoprotein (HDL) cholesterol, thereby adversely influencing the risk profile for cardiovascular disease. Neither does a low fat diet appear to reduce obesity, another risk factor. A modern advice based on published research reads: obese persons should reduce their intake of saturated and trans-fatty acids by lowering the consumption of dairy fat, meat and hardened oils (bakery products and catering products fried in hardened fats), and in addition should consume less products with added sugars and refined starch. Carbohydrates should be provided by fruits, vegetables, leguminous plants and whole-wheat products. Persons of about the ideal weight should replace saturated and trans-fatty acids in their diet by unsaturated plant oils, and products with refined carbohydrates by fruits, vegetables and whole-wheat products.


Frequency of Seaweed Intake and Its Association with Cardiovascular Disease Mortality: The JACC Study.
Seaweed is a popular traditional foodstuff in Asian countries. To our knowledge, few studies have examined the association of seaweed intake with mortality from cardiovascular disease. We examined the association of frequency of seaweed intake with total and specific cardiovascular disease mortality. We examined the association of seaweed intake with mortality from cardiovascular disease among 40,234 men and 55,981 women who participated in the Japan Collaborative Cohort Study for Evaluation of Cancer Risk. Sex-specific hazard ratios for mortality from cardiovascular disease (stroke, stroke subtypes, and coronary heart disease) according to the frequency of seaweed intake were calculated stratified by study area and adjusted for potential cardiovascular risk factors and dietary factors. During the 1,580,996 person-year follow-up, 6,525 cardiovascular deaths occurred, of which 2,820 were due to stroke, and 1,378, to coronary heart disease. Among men, the multivariable analysis showed that participants who ate seaweed almost every day compared with those who never ate seaweed had hazard ratios (95% confidence interval; P for trend) of 0.79 (0.62-1.01; 0.72) for total cardiovascular disease, 0.70 (0.49-0.99; 0.47) for total stroke, 0.69 (0.41-1.16; 0.11) for cerebral infarction. Among women, the multivariable-adjusted hazard ratios were 0.72 (0.55-0.95; 0.001) for total cardiovascular disease, 0.70 (0.46-1.06; 0.01) for total stroke, and 0.49 (0.27-0.90; 0.22) for cerebral infarction. No associations were observed between seaweed intake and risk of intraparenchymal hemorrhage and coronary heart disease among either men or women. We found an inverse association between seaweed intake and cardiovascular mortality among Japanese men and women, especially that from cerebral infarction.


Interpretations of cigarette advertisement warning labels by Philadelphia Puerto Ricans.
This study examined Philadelphia Puerto Ricans' interpretations of the Surgeon General's warnings that appear on cigarette packaging and in advertisements. In-home family focus groups in which participants were asked to comment on magazine cigarette advertisements showed a great variety of interpretations of the legally mandated warning labels. These findings (a) corroborate and add to research in public health and communications regarding the possibility of wide variations in message interpretations and (b) support the call for public health messages to be carefully tested for effectiveness among different social groups. The article's focus on Puerto Ricans addresses the problem of misleading conclusions that can arise from aggregating all Latino subpopulations into one group. The use of a naturalistic setting to examine interpretations of messages about smoking departs from the experimental methods typically used for such research and provides new evidence that even a seemingly straightforward message can be interpreted in multiple ways. Understanding and addressing differences in message interpretation can guide public health campaigns aimed at reducing health disparities.


Oral modified-release formulations in motion: the relationship between gastrointestinal transit and drug absorption.
Oral modified-release dosage forms can be designed with the aim of achieving specific pharmacokinetic profiles, delivering to specific gut localities or reducing the number of drug administrations. Multiple-unit systems, such as pellets, beads or granules, often claim superiority to single-unit modified-release formulations in terms of predictability and reproducibility of behaviour in the gastrointestinal tract. This is an oversimplification and in this review we discuss the effect of the highly variable gastrointestinal transit on the bioperformance of multiple-unit dosage forms, relative to their single-unit counterparts. We examine the sometimes contradictory literature in this area and highlight specific case studies which demonstrate the effect of intestinal transit on dosage form performance and drug absorption.


Regenerative medicine strategies for treating neurogenic bladder.
Neurogenic bladder is a general term encompassing various neurologic dysfunctions of the bladder and the external urethral sphincter. These can be caused by damage or disease. Therapeutic management options can be conservative, minimally invasive, or surgical. The current standard for surgical management is bladder augmentation using intestinal segments. However, because intestinal tissue possesses different functional characteristics than bladder tissue, numerous complications can ensue, including excess mucus production, urinary stone formation, and malignancy. As a result, investigators have sought after alternative solutions. Tissue engineering is a scientific field that uses combinations of cells and biomaterials to encourage regeneration of new, healthy tissue and offers an alternative approach for the replacement of lost or deficient organs, including the bladder. Promising results using tissue-engineered bladder have already been obtained in children with neurogenic bladder caused by myelomeningocele. Human clinical trials, governed by the Food and Drug Administration, are ongoing in the United States in both children and adults to further evaluate the safety and efficacy of this technology. This review will introduce the principles of tissue engineering and discuss how it can be used to treat refractory cases of neurogenic bladder.


An initiative to improve primary prevention of cardiovascular disease in adults with type II diabetes based on the ACC/AHA (2013) and ADA (2016) guidelines.
This project was designed to increase family practice clinicians' adherence to the American College of Cardiology/American Heart Association (2013) and American Diabetes Association (ADA, 2016) clinical practice guidelines for the prevention of cardiovascular disease (CVD) in 40- to 75-year-old adults with type II diabetes (DMII) through interactive education, an electronic alert, and quality improvement sessions on the treatment and management of cholesterol in a family practice primary care setting. A pre- and postretrospective chart review of 159 charts from a family practice clinic was conducted. Included in the chart review were 40- to 75-year-olds with a diagnosis of DMII. Three outcomes were analyzed. Assessment of Atherosclerotic Cardiovascular Disease risk score increased from 0% to 76.7% (X2 (1, 159) = 93.46, p < .001), diet and exercise counseling documentation increased from 85% to 100%, and statin therapy application increased from 55.1% to 72.2% (X2 (1, 159) = 11.55, p = .003) in eligible adults 40-75 years-old with DMII. This initiative helps to build the body of evidence regarding useful interventions to improve primary prevention of CVD by improving clinician adherence to the clinical care guidelines for 40- to 75-year-old adults with DMII.


[How about the uncertainty in the haplotypes in the population-based KORA studies?].
In the KORA surveys, numerous candidate genes in the context of type 2 diabetes, myocardial infarction, atherosclerosis or obesity are under investigation. Current focus is on genotyping single nucleotide polymorphism (SNPs). Haplotypes are also of increasing interest: haplotypes are combinations of alleles within a certain section of one chromosome. Analysing haplotypes in genetic association studies is often more efficient than studying the SNPs separately. A statistical problem in this context is the reconstruction of the phase: genotyping the SNPs determines the alleles of an individual at one particular locus of the DNA, but does not reveal which allele is located on which one of the two chromosomes. This information is required when talking about haplotypes. There are statistical approaches to identify the most likely two haplotypes of an individual given the genotypes. However, a certain error in prognosis is unavoidable. There are also errors in the genotypes. These errors are assumed to be small for one SNP but can accumulate over the SNPs involved in one haplotype and thus can induce further uncertainty in the haplotype. It is therefore the aim of our project to quantify the uncertainties in the haplotypes particularly for genes investigated in the KORA surveys. We conduct computer simulations based on the haplotypes and their frequencies observed in the KORA individuals and compare the results with simulations based on mathematical modelling of the evolutionary process ("coalescent models"). The uncertainties in the haplotypes have an impact on the search for association between genes and disease: an association may not be detected as the haplotype uncertainty obscures the haplotype frequency differences between cases and controls. It is a further aim of our project to elucidate the extent of this problem and to develop strategies for reducing it.


Comparison between Topical and Oral Tranexamic Acid in Management of Traumatic Hyphema.
We sought to determine the efficacy of topical tranexamic acid (5%) in the management of traumatic hyphema. Thirty eyes with gross traumatic hyphema were enrolled in this study. The patients were treated with tranexamic acid (5%) eye drop every 6 hours for 5 days. The main outcome measures were best corrected visual acuity (BCVA), Intra-ocular pressure (IOP), day of clot absorption, and rate of rebleeding. These parameters were evaluated daily for 4 days and thereafter at the 8(th) and 14(th) days after treatment. The patients were also compared with two historical control groups of patients (80 eyes) with traumatic hyphema; the first control group was treated with oral placebo and the other group was treated with oral tranexamic acid at our department. Prior to treatment, the mean logarithm of the minimum angle of resolution (logMAR) BCVA was 0.59±0.62. BCVA was increased to 0.08±0.14 at day 14 (P<0.001) and the mean IOP before treatment was 13.7±3.9 mm Hg, which was reduced to 11.4±1.8 mm Hg at day 14 (P=0.004). Rebleeding occurred in one (3.3%) patient on the 4(th) day post treatment. Comparison between the case group and the other two historical control groups with respect to the rebleeding rate demonstrated statistically significant differences between the case group and the first control group (P=0.008) but no statistically significant differences between the case group and the second control group (P=0.25). Topical tranexamic acid seems promising in the management of traumatic hyphema. However, the small sample size of the present study precludes the conclusion that topical tranexamic acid can replace the oral tranexamic acid.


An Analysis Regarding the Article: Insight Into the Optimal Timing of Percutaneous Coronary Intervention and Transcatheter Aortic Valve Replacement.
Transcatheter aortic valve replacement (TAVR) is an emerging minimally invasive treatment modality that has been shown to reduce mortality significantly compared to medical therapy alone and is superior or noninferior to conventional surgical aortic valve replacement (SAVR) in patients with severe and symptomatic aortic stenosis (AS) deemed to have an intermediate or higher operative risk. AS patients at intermediate surgical risk, although complications of each procedure were different; with vascular complications and paravalvular leak (PVL) being more common in the TAVI group, whereas several other complications were more frequently found in the SAVR group, including new-onset atrial fibrillation, acute kidney injury and life-threatening bleeding. The risk factors associated with TAVR include larger annulus size (based on MSCT), pre-TAVI transvalvular Vmax, and aortic valve calcification; aortic valve calcification was found to be an independent predictor of PVL.


Complete Genome Sequence of Bradyrhizobium japonicum Podophage Paso.
Bradyrhizobium japonicum is a nitrogen-fixing, Gram-negative bacterium that forms a symbiotic relationship with leguminous plants. This announcement describes the isolation and genome annotation of B. japonicum T7-like podophage Paso. Genomic analysis reveals genes that are associated with both the T5 and T7 modes of genomic DNA entry into the host.


Extracting symptoms from free-text responses using ChatGPT among COVID-19 cases in Hong Kong.
To investigate the feasibility and performance of Chat Generative Pretrained Transformer (ChatGPT) in converting symptom narratives into structured symptom labels. We extracted symptoms from 300 deidentified symptom narratives of COVID-19 patients by a computer-based matching algorithm (the standard), and prompt engineering in ChatGPT. Common symptoms were those with a prevalence >10% according to the standard, and similarly less common symptoms were those with a prevalence of 2-10%. The precision of ChatGPT was compared with the standard using sensitivity and specificity with 95% exact binomial CIs (95% binCIs). In ChatGPT, we prompted without examples (zero-shot prompting) and with examples (few-shot prompting). In zero-shot prompting, GPT-4 achieved high specificity (0.947 [95% binCI: 0.894-0.978]-1.000 [95% binCI: 0.965-0.988, 1.000]) for all symptoms, high sensitivity for common symptoms (0.853 [95% binCI: 0.689-0.950]-1.000 [95% binCI: 0.951-1.000]), and moderate sensitivity for less common symptoms (0.200 [95% binCI: 0.043-0.481]-1.000 [95% binCI: 0.590-0.815, 1.000]). Few-shot prompting increased the sensitivity and specificity. GPT-4 outperformed GPT-3.5 in response accuracy and consistent labelling. This work substantiates ChatGPT's role as a research tool in medical fields. Its performance in converting symptom narratives to structured symptom labels was encouraging, saving time and effort in compiling the task-specific training data. It potentially accelerates free-text data compilation and synthesis in future disease outbreaks and improves the accuracy of symptom checkers. Focused prompt training addressing ambiguous descriptions impacts medical research positively.


[Factors associated with global and leisure-time physical activity in the elderly: a health survey in São Paulo (ISA-SP), Brazil].
The purpose of this study was to measure the prevalence of global and leisure-time physical activity and associated factors in the elderly. This was a population-based cross-sectional study covering a multiple-stage sample of 1,950 subjects 60 years or older living in areas of São Paulo State, Brazil. Prevalence of global physical activity (assessed through the short version of the International Physical Activity Questionnaire - IPAQ) was 73.9%, and prevalence of leisure-time physical activity was 28.4%. The results highlight the differences between factors associated with global and leisure-time physical activities. The social groups most prone to overall sedentary lifestyle and especially to lack of leisure-time physical activity should be the main targets of health policies aimed at promoting healthier lifestyles.


Antimicrobial screening of Bridelia, micrantha, Alchormea cordifolia and Boerhavia diffusa.
This report is on the antimicrobial potential of Bridelia micrantha, Alchornea cordifolia and Boerhavia diffusa sourced from traditional healers through an ethnobotanical survey of antiinfective plants in Egbado South in Ogun State, Nigeria. Extracts of B. micrantha and A. cordifolia exhibited significant inhibitory activity against the pathogenic organisms. In some cases, the antibacterial activity was comparable to those of ampicillin and gentamycin. However, only the leaf of A. cordifolia showed reasonable antifungal activity when compared with Trosyd. The study shows that there is justification for the use of these medicinal plants in traditional medicine.


[The prognostic value of eicosanoids in the acute respiratory distress syndrome].
There is a great number of agents involved in the acute respiratory distress syndrome (ARDS) physiopathology, and some of them may have a prognostic value. The objective of the present study has been to analyse the prognostic value of eicosanoids in this syndrome. A prospective study with 21 consecutive ARDS patients admitted to the intensive care unit of a therapy hospital in Barcelona, Spain, was carried out. In the first 48 h of the ARDS diagnosis, at baseline, the plasma levels, (in peripheral arterial and pulmonary arterial samples) of thromboxane B2 (TXB2), prostaglandin F1-alpha) (PGF1-alpha) and leukotriene B4 (LTB4) were analysed by RIA. Simultaneously we measured different pulmonary and systemic hemodynamical variables, as well as the pulmonary gas exchange data. We also studied the venous levels of the same eicosanoids in 17 healthy adults, used as reference. Plasma levels of eicosanoids in the ARDS patients were higher than reference subjects (p < 0.05). No differences were observed between systemic arterial and pulmonary arterial values. From all the eicosanoids, only LTB4, (in both systemic arterial and pulmonary blood), was correlated with LIS (r = 0.49, p < 0.05; and r = 0.45, p < 0.05, respectively). Patients who did not survive presented a lower systemic-pulmonary arterial gradient of eicosanoids levels than survivors (-1.27 vs -0.10 ng/ml; p < 0.01). In our ARDS patients only LTB4 plasma levels correlated with the severity of respiratory failure. Patients who did not survive presented a lower LTB4 gradient than survivors.


Immunodiagnosis of tumors in vivo using radiolabeled monoclonal antibody A2B5.
Recently a murine monoclonal antibody (A2B5) has been described that reacts with a membrane associated GQ ganglioside common to peptide secreting normal cells and tumors. In vitro binding data demonstrated the presence of this ganglioside on neurons, adrenal medulla, and pancreatic islets, along with neuroendocrine tumors such as insulinomas, pheochromocytomas, melanomas and neuroblastomas. Negative binding has previously been shown for tissue sections from liver, kidney, colon, lung, stomach, and tumors not derived from the neural crest. Because of the specificity at A2B5 in vitro, this monoclonal antibody was labeled with 131I for in vivo tumor localization studies. Daily radionuclear scans were obtained in 5 KX rats bearing the radiation induced rat insulinoma with disappearance of the label from the blood pool and concentration in the tumor so that by the fourth day, the only activity present by scan was in the insulinoma. Tissue-counting data showed tumor/blood ratios (av +/- SE, 1.29 +/- 0.25) of A2B5 activity two to ten times the average activity found in other organs (0.28 +/- 0.05). No tumor concentration of the control nonspecific monoclonal antibody P3X63 was evident (0.27 +/- 0.04). In addition A2B5 also localized to five different human melanoma cells lines grown in nude mice with high tumor/blood levels (1.04 +/- 0.27) compared to normal tissues (0.32 +/- 0.05) (P = .0005), while no localization is seen in nudes carrying osteosarcomas, colon, bladder, and renal cell carcinomas. In addition antibody A2B5 did not concentrate in any normal tissue though the antigen is present on several. The finding that A2B5 reacts across species lines (mouse, rat, man) lends itself to obvious diagnostic and therapeutic possibilities.


Surfactant function affected by airway inflammation and cooling: possible impact on exercise-induced asthma.
Pulmonary surfactant maintains patency of narrow conducting airways. An inflammation, with a leakage of plasma proteins into the airway lumen, causes surfactant to lose some of this ability. Will a lowering of temperature aggravate the deteriorating effect of an inflammation? Calf lung surfactant extract (CLSE) with proteins added was studied with a capillary surfactometer (CS) at temperatures of 25-42 degrees C. BALB/c mice were infected with respiratory syncytial virus (RSV). Six days later the lungs were lavaged and the surfactant in the lavage fluid was studied with the CS at temperatures of 25-42 degrees C. Lavage fluid from allergen challenged asthmatics was examined for its content of surfactant inhibitors at reduced temperatures. It was shown that CLSE with proteins gradually lost its ability to maintain patency as the temperature was lowered. Lavage fluid from the RSV infected mice showed a similar dysfunction at low temperatures. Lavage fluid from the airways of human asthmatics, when challenged with antigen but not with saline, contained agents inhibiting surface activity, particularly at reduced temperatures. Airway inflammation causes surfactant to lose its ability to maintain patency, particularly as the temperature is reduced. That might be a reason for the increased airway resistance observed in asthma patients hyperventilating in cold weather.


Linking Engineered Cells to Their Digital Twins: A Version Control System for Strain Engineering.
As DNA sequencing and synthesis become cheaper and more easily accessible, the scale and complexity of biological engineering projects is set to grow. Yet, although there is an accelerating convergence between biotechnology and digital technology, a deficit in software and laboratory techniques diminishes the ability to make biotechnology more agile, reproducible, and transparent while, at the same time, limiting the security and safety of synthetic biology constructs. To partially address some of these problems, this paper presents an approach for physically linking engineered cells to their digital footprint-we called it digital twinning. This enables the tracking of the entire engineering history of a cell line in a specialized version control system for collaborative strain engineering via simple barcoding protocols.


Competitive Interactions between PIRT, the Cold Sensing Ion Channel TRPM8, and PIP2 Suggest a Mechanism for Regulation.
TRPM8 is a member of the transient receptor potential ion channel family where it functions as a cold and pain sensor in humans and other higher organisms. Previous studies show that TRPM8 requires the signaling phosphoinositide lipid PIP2 to function. TRPM8 function is further regulated by other diverse mechanisms, including the small modulatory membrane protein PIRT (phosphoinositide regulator of TRP). Like TRPM8, PIRT also binds PIP2 and behavioral studies have shown that PIRT is required for normal TRPM8-mediated cold-sensing. To better understand the molecular mechanism of PIRT regulation of TRPM8, solution nuclear magnetic resonance (NMR) spectroscopy was used to assign the backbone resonances of full-length human PIRT and investigate the direct binding of PIRT to PIP2 and the human TRPM8 S1-S4 transmembrane domain. Microscale thermophoresis (MST) binding studies validate the NMR results and identify a competitive PIRT interaction between PIP2 and the TRPM8 S1-S4 domain. Computational PIP2 docking to a human TRPM8 comparative model was performed to help localize where PIRT may bind TRPM8. Taken together, our data suggest a mechanism where TRPM8, PIRT, and PIP2 form a regulatory complex and PIRT modulation of TRPM8 arises, at least in part, by regulating local concentrations of PIP2 accessible to TRPM8.


Professional resocialization of post-RN baccalaureate students by distance education.
Distance education is being used to increase access to post-RN baccalaureate education. One of the objectives of degree programs is the professional resocialization of post-RN students. Because of the limited contacts with the institution, faculty, and student peers, the effectiveness of professional resocialization by distance education may be questioned. In this qualitative study, the responses of 24 students who had taken distance courses in Nursing Issues from four universities were analyzed for evidence of professional resocialization. Two of the universities used a group-oriented approach to distance education; two used an individual approach. Responses of students from the four schools and the two approaches were similar. All reported more awareness of issues in nursing and most reported attitude changes reflecting a more professional orientation. Among the changes were attitudes toward baccalaureate entry to practice, nursing theory and research, and the place of nurses and nursing in society. Despite the limitations of distance education, distance courses can be influential in the professional resocialization of post-RN students.


[Work, cardiopathy, and sports].
Heart disease is the main cause of early disability and premature death in Europe. Regular physical activity may prevent heart disease, diabetes, ictus, and obesity. Nevertheless, a certain resistance to a dynamic lifestyle, lack of free time, lack of motivation and other factors are frequently encountered. To stress the importance of physical activity in the prevention of cardiovascular disease. A literature review of the main risk factors for cardiovascular disease was carried out. It has been shown that physical exercise is beneficial to the cardiovascular apparatus and to the bones and joints, by improving some metabolic parameters. Recent studies have shown that a personalized, moderate physical activity should be suggested for primary and secondary prevention, in particular for subjects with coronary artery disease and left ventricular dysfunction. An increase in sports activity and physical exercise at work and at school is recommended so as to improve quality of life and promote home rehabilitation. Some promising experience has already been made among workers and their families with excellent results, as shown by a follow-up period of three years.


IMG-ABC: A Knowledge Base To Fuel Discovery of Biosynthetic Gene Clusters and Novel Secondary Metabolites.
In the discovery of secondary metabolites, analysis of sequence data is a promising exploration path that remains largely underutilized due to the lack of computational platforms that enable such a systematic approach on a large scale. In this work, we present IMG-ABC (https://img.jgi.doe.gov/abc), an atlas of biosynthetic gene clusters within the Integrated Microbial Genomes (IMG) system, which is aimed at harnessing the power of "big" genomic data for discovering small molecules. IMG-ABC relies on IMG's comprehensive integrated structural and functional genomic data for the analysis of biosynthetic gene clusters (BCs) and associated secondary metabolites (SMs). SMs and BCs serve as the two main classes of objects in IMG-ABC, each with a rich collection of attributes. A unique feature of IMG-ABC is the incorporation of both experimentally validated and computationally predicted BCs in genomes as well as metagenomes, thus identifying BCs in uncultured populations and rare taxa. We demonstrate the strength of IMG-ABC's focused integrated analysis tools in enabling the exploration of microbial secondary metabolism on a global scale, through the discovery of phenazine-producing clusters for the first time in Alphaproteobacteria. IMG-ABC strives to fill the long-existent void of resources for computational exploration of the secondary metabolism universe; its underlying scalable framework enables traversal of uncovered phylogenetic and chemical structure space, serving as a doorway to a new era in the discovery of novel molecules. IMG-ABC is the largest publicly available database of predicted and experimental biosynthetic gene clusters and the secondary metabolites they produce. The system also includes powerful search and analysis tools that are integrated with IMG's extensive genomic/metagenomic data and analysis tool kits. As new research on biosynthetic gene clusters and secondary metabolites is published and more genomes are sequenced, IMG-ABC will continue to expand, with the goal of becoming an essential component of any bioinformatic exploration of the secondary metabolism world.


Efficacy and safety of TNF antagonists in ocular sarcoidosis: data from the French registry STAT.
Backgroung: This study investigated the efficacy and safety of TNF antagonists in sarcoid uveitis in unselected cases. Design: This is a multicentre study on patients with sarcoidosis who received TNF antagonists in pneumology and internal medicine departments in France. We present here the subgroup of patients with biopsy-proven sarcoid uveitis included in the nationwide registry STAT (Sarcoidosis treated with TNF AnTagonists). Results: Among the 132 patients included in this multicenter study, 18 patients with refractory uveitis were treated as a first-line TNF antagonist with infliximab (n=14), adalimumab (n=3) and certolizumab (n=1). Before anti-TNF initiation, the median duration of sarcoidosis was 42 months and 83% of the patients have been treated with at least one immunosuppressive drug. Six patients switched for a second-line TNF antagonist. After a mean time under treatment of 29 months, the treatment resulted in a significant decrease of the ophthalmic extrapulmonary Physician Organ Severity Tool (ePOST) (mean score: 4.2 vs. 2.6) scores and a steroid sparing effect (29.4±20.7 vs. 6.2±5.2 mg/d). Overall, the ophthalmic response, either complete or partial, was 67%. Nine patients (50%) presented adverse events, including severe infectious complications in 5 patients, which required anti-TNF treatment interruption in 6 cases (33%). Among the 7 responder patients who discontinued anti-TNF therapy, 71% relapsed. Finally, 12 patients (67%) could continue TNF antagonist treatment. Conclusions: TNF antagonists were efficient in 67% of biopsy-proven refractory sarcoid uveitis. Severe adverse events, mainly infectious complications, were frequent. The high frequency of relapses after anti-TNF-α discontinuation requires a close patient follow-up thereafter. (Sarcoidosis Vasc Diffuse Lung Dis 2017; 34: 74-80).


Current and future trade in livestock products.
Rising per capita consumption, economic growth, and urbanisation, particularly in developing countries, have been driving an increased global demand for food. These changing socio-economic trends, which have greatly influenced changes in dietary patterns globally and, more specifically, have increased consumption of livestock products in developing countries, are expected to endure and to place new pressures on livestock-sector infrastructure and the delivery of veterinary services. This paper summarises current trade in meat and presents plausible projections for the future. It highlights the impact of animal disease on trade and considers the effect of ongoing disease outbreaks, particularly the outbreaks of African swine fever and COVID-19, on current and future trade dynamics. The authors analysed published statistics on the demand for, and international trade in, livestock products at national and regional levels and made projections of the same up to 2050, generated from an integrated model of the global agricultural and food system. The resulting analyses identified patterns of trade consistent with growing populations, increasing incomes and changing diets in developing countries. The analyses also pointed to slow expansion of livestock production, and the impacts of countries' disease status on livestock trade. For most of the livestock products analysed, economic model projections indicate increased consolidation of production and exports among a few countries. Marked increases in the trade in livestock products suggest a changing role for Veterinary Services in facilitating trade and extension in the years to come.


Prevalence of Cardiovascular Diseases in South Asians: Scrutinizing Traditional Risk Factors and Newly Recognized Risk Factors Sarcopenia and Osteopenia/Osteoporosis.
One of the primary reasons for complications and death worldwide are cardiovascular diseases (CVDs), with a death toll of approximately 18 million per year. CVDs include cardiomyopathy, hypertension, ischemic heart disease, coronary heart disease, myocardial infarction, heart attack, hearth failure, etc. Over 80% of the CVD mortality is recorded from lower and middle-income countries. Records from the past decade have highlighted the increase of CVDs among the South Asian populations, and the prime purpose of the review is to jot down the reasons for the steep spike in CVDs. Studies analyzing the causative factors for the increase of CVDs in South Asians are still to be verified. Apart from known predisposing and lifestyle factors, other emerging risk factors associated with CVDs, namely the musculoskeletal diseases sarcopenia and osteopenia, should be tracked to tackle research gaps in upcoming analyses. This requires loads of scientific efforts. With proper monitoring, the raising alarm that the CVD burden generates can be reduced. This review discusses the already established signs and recognizes important clues to the emerging etiology of CVDs in the Asian population and prevention measures to keep it at bay.


Leishmania major drives host phagocyte death and cell-to-cell transfer depending on intracellular pathogen proliferation rate.
The virulence of intracellular pathogens relies largely on the ability to survive and replicate within phagocytes but also on release and transfer into new host cells. Such cell-to-cell transfer could represent a target for counteracting microbial pathogenesis. However, our understanding of the underlying cellular and molecular processes remains woefully insufficient. Using intravital 2-photon microscopy of caspase-3 activation in the Leishmania major-infected (L. major-infected) live skin, we showed increased apoptosis in cells infected by the parasite. Also, transfer of the parasite to new host cells occurred directly without a detectable extracellular state and was associated with concomitant uptake of cellular material from the original host cell. These in vivo findings were fully recapitulated in infections of isolated human phagocytes. Furthermore, we observed that high pathogen proliferation increased cell death in infected cells, and long-term residency within an infected host cell was only possible for slowly proliferating parasites. Our results therefore suggest that L. major drives its own dissemination to new phagocytes by inducing host cell death in a proliferation-dependent manner.


GABAergic interneurons excite neonatal hippocampus in vivo.
GABAergic interneurons are proposed to be critical for early activity and synapse formation by directly exciting, rather than inhibiting, neurons in developing hippocampus and neocortex. However, the role of GABAergic neurons in the generation of neonatal network activity has not been tested in vivo, and recent studies have challenged the excitatory nature of early GABA. By locally manipulating interneuron activity in unanesthetized neonatal mice, we show that GABAergic neurons are excitatory in CA1 hippocampus at postnatal day 3 (P3) and are responsible for most of the spontaneous firing of pyramidal cells at that age. Hippocampal interneurons become inhibitory by P7, whereas visual cortex interneurons are already inhibitory by P3 and remain so throughout development. These regional and age-specific differences are the result of a change in chloride reversal potential, because direct activation of light-gated anion channels in glutamatergic neurons drives CA1 firing at P3, but silences it at P7 in CA1, and at all ages in visual cortex. This study in the intact brain reveals that GABAergic interneuron excitation is essential for network activity in neonatal hippocampus and confirms that visual cortical interneurons are inhibitory throughout early postnatal development.


Distribution pattern of dissolved organic matter in pore water of sediments from three typical areas of western Lake Taihu and its environmental implications.
The migration, transformation, and accumulation of dissolved organic matter (DOM) in pore water of sediment cores play a pivotal role in lacustrine carbon cycling. In order to understand the dynamics of DOM in the sediments of large shallow eutrophic lakes, we examined the vertical profiles of DOM and the benthic fluxes of dissolved organic carbon (DOC) in sediment cores located in algae accumulated, dredged, and central areas of eutrophic Lake Taihu, China. Optical properties showed the significant influence of terrestrial inputs on the DOM components of pore water in the algae accumulated area but an abundant accumulation of autochthonous DOM in the central area. The benthic fluxes of DOC ranging from -458.2 to -139.4 mg·m-2·d-1 in the algae accumulated area displayed an opposite diffusion direction to the other two areas. The flux ranges of 9.5-31.2 mg·m-2·d-1 in the dredged area and 14.6-48.0 mg·m-2·d-1 in the central area were relatively smaller than those in the previously reported lake ecosystems with low trophic levels. Dredging engineering disturbed the pre-dredging distribution patterns of DOM in sediment cores. The deposition, accumulation, and transformation of massive algae scums in eutrophic lakes probably promoted the humification degree of sediments.


Elevated levels of anticardiolipin antibodies and epilepsy in lupus patients.
To examine the association between anticardiolipin (aCL) antibodies and epilepsy, we investigated the serum titers of aCL antibodies in a total 252 systemic lupus erythematosus (SLE) patients recruited in a prospective study. Twenty-one cases with epilepsy which were not attributable to any causes other than SLE were identified after being followed-up for five years. The clinical manifestations were recorded and blood samples were tested for the presence of aCL antibodies (IgG, IgM and IgA isotypes). Among 21 patients with epilepsy, 12 (57.1%), 2 (9.5%) and 2 (9.5%), respectively, had elevated baseline serum levels of IgG, IgM and IgA aCL antibodies. There was a dose-response relationship between risk of seizure and the baseline serum level of aCL antibodies (P < 0.01). The odds ratio of developing seizure were 3.7 for those who had a high level of aCL antibodies compared with those without a detectable level of aCL antibodies as the referent. Our results indicate that epilepsy as a primary neuropsychiatric event among lupus patients is associated with a high titer of aCL antibodies.


Protein alignment algorithms with an efficient backtracking routine on multiple GPUs.
Pairwise sequence alignment methods are widely used in biological research. The increasing number of sequences is perceived as one of the upcoming challenges for sequence alignment methods in the nearest future. To overcome this challenge several GPU (Graphics Processing Unit) computing approaches have been proposed lately. These solutions show a great potential of a GPU platform but in most cases address the problem of sequence database scanning and computing only the alignment score whereas the alignment itself is omitted. Thus, the need arose to implement the global and semiglobal Needleman-Wunsch, and Smith-Waterman algorithms with a backtracking procedure which is needed to construct the alignment. In this paper we present the solution that performs the alignment of every given sequence pair, which is a required step for progressive multiple sequence alignment methods, as well as for DNA recognition at the DNA assembly stage. Performed tests show that the implementation, with performance up to 6.3 GCUPS on a single GPU for affine gap penalties, is very efficient in comparison to other CPU and GPU-based solutions. Moreover, multiple GPUs support with load balancing makes the application very scalable. The article shows that the backtracking procedure of the sequence alignment algorithms may be designed to fit in with the GPU architecture. Therefore, our algorithm, apart from scores, is able to compute pairwise alignments. This opens a wide range of new possibilities, allowing other methods from the area of molecular biology to take advantage of the new computational architecture. Performed tests show that the efficiency of the implementation is excellent. Moreover, the speed of our GPU-based algorithms can be almost linearly increased when using more than one graphics card.


[Preparation of hybrid proteins consisting of human interleukin-2 and the cytotoxic A-subunit of Shigella toxin].
Recombinant plasmids providing the synthesis of chimeric proteins consisting of amino acid sequences of human interleukin-2 (IL-2) and Shiga toxin cytotoxic A-subunit (ILA and AIL chimeric toxins) were constructed. The ILA and AIL chimeric toxins were shown to inhibit protein synthesis in the rabbit reticulocytes cell-free system. These chimeric toxins displayed two opposite activities of the constituent parts of their molecules on T-lymphocytes from the peripheral blood of healthy volunteers. Hybrid protein AIL (approximately 10(-6) g/ml) has caused the most significant depression of T-lymphoblast proliferation.


Biologic classification as an alternative to anatomic staging for clinically localized prostate cancer: a proposal based on patients treated with external beam radiotherapy.
The prognostic significance of clinical stage in patients with prostate cancer who are treated with external beam radiotherapy is unclear. This study evaluates multiple pretreatment factors, including clinical stage, to determine which are the best prognostic factors, and develops a classification system based on these factors. All 249 evaluable patients with clinically localized adenocarcinoma of the prostate treated with definitive conformal external beam radiotherapy without androgen deprivation at our institution between 1989 and 1993 were analyzed. Clinical stage, serum PSA level, Gleason score, race, and history of transurethral resection of the prostate (TURP) were evaluated for their ability to predict biochemical disease-free survival (BDFS). Factors predictive of BDFS were then used to construct a classification system. The classification system was then analyzed for its ability to predict BDFS, distant metastases, local recurrence, and clinical disease free survival in univariate and multivariate analyses. Median follow-up was 27 months. Gleason score and PSA predicted BDFS in multivariate analysis (both P <0.0001), whereas clinical stage, race, and history of a TURP did not. These two biologic factors were combined into a four-level classification system. This classification system was analyzed together with Gleason score and PSA and was found to be the only predictor of BDFS on multivariate analysis (P <0.0001). In addition, this classification system was the only predictor of distant metastases in multivariate analysis (P <0.0001). The classification system derived herein based on the biologic factors of Gleason score and serum PSA levels is the sole predictor of distant metastases and biochemical recurrence for patients treated with definitive conformal external beam radiotherapy for clinically localized prostate cancer. This classification system may be useful when comparing competing therapies and stratifying patients in clinical trials, but requires validation from other institutions and other therapies prior to its widespread use.


CO diffusion as a re-orientation mechanism in the NaY zeolite.
Our work is devoted to DFT calculations of the relative rotational and diffusional barriers for CO motions in zeolite NaY. The diffusion jump of CO adsorbed in NaY from NaII to Na'II has been confirmed as the favored way for CO re-coordination via either the C or the O atom to the Na cations instead of the CO rotation, hence explaining the mechanism which is responsible for the CO exchange between different positions and the changes in the intensities of the vibrational IR spectra. The fine structure of the vibrational C-O bands is explained by the different CO locations of adsorbed mono- and dicarbonyl species. The calculated activation energy of intra-cage CO diffusion from NaII-CO to Na'II-OC matches the respective experimental barrier observed in the NaX zeolite.


Investigation of Taste-Related Compounds and Antioxidative Profiles of Retorted Samgyetang Made from Fresh and Dried Cordyceps militaris Mushrooms.
This study was performed to investigate the effects of taste-related compounds and antioxidatve profiles of retorted samgyetang made from fresh and dried Cordyceps militaris (C. militaris) mushrooms. A total of 48 carcasses were prepared from commercial broilers (CB; Ross, 4 weeks old) and randomly distributed into eight different treatments. Each treatment group consisted of 6 chicken carcasses made with the addition of broth in different condition and concentration of C. militaris mushrooms. The addition concentration was based on the broth volume (v/w) under either fresh or dried conditions ranging from 0% as a control to 1%, 2%, and 3% of C. militaris mushrooms. C. militaris mushrooms contributed to an improvement of meat tenderness and the antioxidative profile that led to a greater suppression of lipid oxidation. The addition of C. militaris mushrooms at 2% could also enrich the flavor and taste-related compounds, particularly the increase in 5'-AMP and umami-related free amino acid compounds, L-aspartic acid and L-glutamic acid. Different addition forms of C. militaris mushrooms, particularly fresh or dried mushrooms, had only small effects on bioactive compounds, where the dried addition could possibly enrich samgyetang broth with higher cordycepin and adenosine contents than the fresh addition. Besides, the addition of C. militaris mushrooms in the dried form could also contribute to a higher antioxidative profile. Eventually, the addition of C. militaris mushrooms with a minimum addition of 2% contributed to an improvement of meat quality, antioxidative profile and flavor improvement of samgyetang.


"Development of Fixed Dose Combination Products" Workshop Report: Considerations of Gastrointestinal Physiology and Overall Development Strategy.
The gastrointestinal (GI) tract is one of the most popular and used routes of drug product administration due to the convenience for better patient compliance and reduced costs to the patient compared to other routes. However, its complex nature poses a great challenge for formulation scientists when developing more complex dosage forms such as those combining two or more drugs. Fixed dose combination (FDC) products are two or more single active ingredients combined in a single dosage form. This formulation strategy represents a novel formulation which is as safe and effective compared to every mono-product separately. A complex drug product, to be dosed through a complex route, requires judicious considerations for formulation development. Additionally, it represents a challenge from a regulatory perspective at the time of demonstrating bioequivalence (BE) for generic versions of such drug products. This report gives the reader a summary of a 2-day short course that took place on the third and fourth of November at the Annual Association of Pharmaceutical Scientists (AAPS) meeting in 2018 at Washington, D.C. This manuscript will offer a comprehensive view of the most influential aspects of the GI physiology on the absorption of drugs and current techniques to help understand the fate of orally ingested drug products in the complex environment represented by the GI tract. Through case studies on FDC product development and regulatory issues, this manuscript will provide a great opportunity for readers to explore avenues for successfully developing FDC products and their generic versions.


Kinetics of retinal lipoprotein precipitation and elimination after closure of subretinal new vessels.
To study the kinetics of retinal precipitation and elimination of lipoprotein in relation to photodynamic therapy of subretinal new vessels. This was a retrospective observational study of subretinal precipitate before and after photodynamic therapy for subfoveal new vessels secondary to age-related macular degeneration in 14 eyes demonstrating precipitate at one or more visits, using digital red-free gray-scale fundus photography, intravenous fluorescein angiography, optical coherence tomography, and morphometric mapping. In 14 eyes of 14 patients, the area covered by lipoprotein precipitate increased from a mean of 0.54 optic disc areas (range, 0-2.61) before treatment to a mean of 0.65 optic disc areas (range, 0.01-3.04) at 45 d after treatment (P = 0.18). Eyes with a serous detachment at baseline, before the initial treatment (n = 10) demonstrated more treatment-related precipitation than eyes without a serous detachment at baseline (n = 4; P = 0.034). Two eyes with a large amount of precipitate and a long follow-up demonstrated net elimination of precipitate from approximately 100 days after the initial treatment, with uniexponential precipitate half-lives of 43 and 32 days. Photodynamic therapy for subfoveal new vessels may be associated with retinal precipitation of lipoprotein, presumably because early extraction of water and salts from the subretinal fluid increases the concentration of leaked plasma proteins. Despite reperfusion of the subretinal vessels, macromolecular leakage appears to cease within 100 days, indicating that functional maturation of the new vessels has occurred, with an associated decrease in pore size.


Mechanistic and biological characterisation of novel N5-substituted paullones targeting the biosynthesis of trypanothione in Leishmania.
Trypanothione synthetase (TryS) produces N1,N8-bis(glutathionyl)spermidine (or trypanothione) at the expense of ATP. Trypanothione is a metabolite unique and essential for survival and drug-resistance of trypanosomatid parasites. In this study, we report the mechanistic and biological characterisation of optimised N5-substituted paullone analogues with anti-TryS activity. Several of the new derivatives retained submicromolar IC50 against leishmanial TryS. The binding mode to TryS of the most potent paullones has been revealed by means of kinetic, biophysical and molecular modelling approaches. A subset of analogues showed an improved potency (EC50 0.5-10 µM) and selectivity (20-35) against the clinically relevant stage of Leishmania braziliensis (mucocutaneous leishmaniasis) and L. infantum (visceral leishmaniasis). For a selected derivative, the mode of action involved intracellular depletion of trypanothione. Our findings shed light on the molecular interaction of TryS with rationally designed inhibitors and disclose a new set of compounds with on-target activity against different Leishmania species.


Differential expression of BCL-2 oncoprotein and Fas antigen on normal peripheral blood and leukemic bone marrow cells. A flow cytometric analysis.
Fas antigen (Ag) has recently been identified as the putative surface molecule capable of transducing apoptotic signals into cells. Alterations in the expression of proto-oncogene bcl-2 have been implicated in the regulation of apoptosis. By employing a monoclonal antibody to bcl-2 protein (124 clone) and to Fas Ag (UB2 clone) the expression of these molecules was analyzed at flow cytometry on bone marrow (BM) and peripheral blood (PB) samples from patients suffering from different lymphoid and myeloid leukemic diseases (27 acute non-lymphocytic leukemia [ANLL]; 14 acute lymphocytic leukemia [ALL]; 19 B-cell chronic lymphocytic leukemia [CLL]; 2 Ph1+ chronic myeloid leukemia [CML]; one CD8+ T-cell chronic lymphoproliferative disorders). Results were compared with those observed on normal PB leukocytes and BM B-cell precursors from patients with non-neoplastic hematological disorders. Fas Ag was constitutively expressed by both monocytes and neutrophils, while lymphocytes expressed bcl-2 with no difference between B and T cell subsets. Interestingly, bcl-2 expression was always absent on neutrophils. When dealing with ANLL patients, a relatively low bcl-2 and high Fas Ag phenotype characterized subtypes with granulocytic (M2) or promyelocytic (M3) differentiation. This observation was confirmed in a small number of patients for whom bcl-2 levels were quantified as antibody binding capacity (ABC) in molecules/cell. Leukemic cells from patients with ALL constitutively expressed bcl-2, the pattern of this expression being quantitatively lower than that of immature B-cell precursors. Finally, high bcl-2 and low Fas Ag expression represented a crucial part of the B-cell CLL immunophenotype. Although based on a small number of patient and control samples, our results suggest that bcl-2 and Fas Ag are coordinately expressed on normal PB leukocytes. Fas Ag is expressed at low levels on B-CLL cells, generally considered long-surviving cells. The relatively lower bcl-2-expression detected in both M2 and M3 subtypes may explain, at least in part, the higher remission rates obtained in these forms of ANLL than in other less differentiated morphological variants.


An unusual way to improve lung function in congenital myopathies: the power of singing.
Congenital myopathies (CMs) are a clinically and genetically heterogeneous group of disorders characterized by early onset weakness, hypotonia and characteristic structural abnormalities in muscle fibres. Hypotonia and weakness can be present at birth or appear in infancy, and a static or slowly progressive clinical course may present with muscle weakness, loss of spontaneous movement, involuntary muscle activity, and muscle atrophy. Often patients develop a restrictive syndrome and respiratory failure and require respiratory support In our case, we described lung improvement and respiratory muscle training due to singing in a young patient, affected by CMs with a poor adherence to non-invasive mechanical ventilation.


Day-to-day deviations in sleep parameters and biological aging: Findings from the NHANES 2011-2014.
The majority of the previous research has focused on the impact of average sleep parameters on longevity. In this study, we aimed to investigate the associations of day-to-day deviations in sleep parameters with biological ages among 6052 adults participating in the 2011-2014 waves of the US National Health and Nutrition Examination Survey. Sleep parameters, including sleep duration, efficiency, midpoint, and day-to-day deviations in sleep parameters, including standard deviation of sleep duration (sleep variability), standard deviation of sleep midpoint (sleep irregularity), catch-up sleep, and social jetlag, were obtained from 4 to 7 days of 24-h accelerometer recording. We used physiological data to compute measurements of biological aging according to 3 published algorithms: PhenoAge, Klemera-Doubal method Biological Age, and homeostatic dysregulation. After adjustment of multiple covariates, we observed that all parameters of day-to-day deviations in sleep were significantly associated with biological aging with larger sleep variability, larger sleep irregularity, more catch-up sleep, and more social jetlag linked with more advanced biological aging. The significant associations of sleep irregularity, catch-up sleep, and social jetlag with biological aging indices remained even after adjustment for sleep duration, efficiency, and midpoint. In this study, we found that day-to-day deviations in sleep parameters are independently associated with biological aging in US general population. Since day-to-day deviation in sleep is a modifiable behavioral factor, our finding suggests that intervention aiming at increasing regularity in sleep patterns may be a novel approach for extending a healthy life span.


Nutritional and botanical interventions to assist with the adaptation to stress.
Prolonged stress, whether a result of mental/emotional upset or due to physical factors such as malnutrition, surgery, chemical exposure, excessive exercise, sleep deprivation, or a host of other environmental causes, results in predictable systemic effects. The systemic effects of stress include increased levels of stress hormones such as cortisol, a decline in certain aspects of immune system function such as natural killer cell cytotoxicity or secretory-IgA levels, and a disruption of gastrointestinal microflora balance. These systemic changes might be a substantial contributor to many of the stress-associated declines in health. Based on human and animal research, it appears a variety of nutritional and botanical substances - such as adaptogenic herbs, specific vitamins including ascorbic acid, vitamins B1 and B6, the coenzyme forms of vitamin B5 (pantethine) and B12 (methylcobalamin), the amino acid tyrosine, and other nutrients such as lipoic acid, phosphatidylserine, and plant sterol/sterolin combinations - may allow individuals to sustain an adaptive response and minimize some of the systemic effects of stress.


Hybrid intravascular imaging: recent advances, technical considerations, and current applications in the study of plaque pathophysiology.
Cumulative evidence from histology-based studies demonstrate that the currently available intravascular imaging techniques have fundamental limitations that do not allow complete and detailed evaluation of plaque morphology and pathobiology, limiting the ability to accurately identify high-risk plaques. To overcome these drawbacks, new efforts are developing for data fusion methodologies and the design of hybrid, dual-probe catheters to enable accurate assessment of plaque characteristics, and reliable identification of high-risk lesions. Today several dual-probe catheters have been introduced including combined near infrared spectroscopy-intravascular ultrasound (NIRS-IVUS), that is already commercially available, IVUS-optical coherence tomography (OCT), the OCT-NIRS, the OCT-near infrared fluorescence (NIRF) molecular imaging, IVUS-NIRF, IVUS intravascular photoacoustic imaging and combined fluorescence lifetime-IVUS imaging. These multimodal approaches appear able to overcome limitations of standalone imaging and provide comprehensive visualization of plaque composition and plaque biology. The aim of this review article is to summarize the advances in hybrid intravascular imaging, discuss the technical challenges that should be addressed in order to have a use in the clinical arena, and present the evidence from their first applications aiming to highlight their potential value in the study of atherosclerosis.


Evaluation of Community-Level Vulnerability and Racial Disparities in Living Donor Kidney Transplant.
Living donor kidney transplant (LDKT) is the ideal treatment for end-stage kidney disease, but racial disparities in LDKT have increased over the last 2 decades. Recipient clinical and social factors do not account for LDKT racial inequities, although comprehensive measures of community-level vulnerability have not been assessed. To determine if racial disparities persist in LDKT independent of community-level vulnerability. This retrospective, multicenter, cross-sectional study included data from 19 287 adult kidney-only transplant recipients in the Scientific Registry of Transplant Recipients. The study included individuals who underwent transplant between January 1 and December 31, 2018. Recipient race and the 2018 US Centers for Disease Control and Prevention Social Vulnerability Index (SVI). Census tract-level SVI data were linked to census tracts within each recipient zip code. The median SVI measure among the census tracts within a zip code was used to describe community-level vulnerability. Kidney transplant donor type (deceased vs living). Modified Poisson regression was used to evaluate the association between SVI and LDKT, and to estimate LDKT likelihood among races, independent of community-level vulnerability and recipient-level characteristics. Among 19 287 kidney transplant recipients, 6080 (32%) received LDKT. A total of 11 582 (60%) were male, and the median (interquartile range) age was 54 (43-63) years. There were 760 Black LDKT recipients (13%), 4865 White LDKT recipients (80%), and 455 LDKT recipients of other races (7%; American Indian, Asian, multiracial, and Pacific Islander). Recipients who lived in communities with higher SVI (ie, more vulnerable) had lower likelihood of LDKT compared with recipients who lived in communities with lower SVI (ie, less vulnerable) (adjusted relative risk [aRR], 0.97; 95% CI, 0.96-0.98; P < .001). Independent of community-level vulnerability, compared with White recipients, Black recipients had 37% lower likelihood (aRR, 0.63; 95% CI, 0.59-0.67; P < .001) and recipients of other races had 24% lower likelihood (aRR, 0.76; 95% CI, 0.70-0.82; P < .001) of LDKT. The interaction between SVI and race was significant among Black recipients, such that the disparity in LDKT between Black and White recipients increased with greater community-level vulnerability (ratio of aRRs, 0.67; 95% CI, 0.51-0.87; P = .003). Community-level vulnerability is associated with access to LDKT but only partially explains LDKT racial disparities. The adverse effects of living in more vulnerable communities were worse for Black recipients. The interaction of these constructs is worrisome and suggests evaluation of other health system factors that may contribute to LDKT racial disparities is needed.


Paramedic decisions with placement of out-of-hospital intravenous lines.
To determine the incidence of unused out-of-hospital intravenous line (IV) placements, we prospectively studied IV placement in emergency medical services (EMS) patients. Unused IV placement was defined as any patient having an EMS initiated IV that was not used for fluid bolus or medication administration in the field or in the emergency department (ED). Data were analyzed on placement and use of IV lines in the field and in the ED, transport time, years of paramedic practice, and paramedic student presence. Of 290 patients, 165 had an IV initiated (147) or attempted (18). Twenty-nine percent (84 of 290) of the patients received an unused EMS IV. One hundred twenty-five patients had no IV initiated by EMS. Seven subsequently had an IV started and used in the ED, for an undertreatment rate of 2.4% (7 of 290). The presence of a paramedic student increased the odds of an unused IV 1.4 (95% CI, 1.1 to 2.0). IVs are frequently started and not used.


A prospective study of early tactile stimulation after digital nerve repair.
Forty-nine patients with 65 digital nerve injuries were randomized into two groups after nerve repair. Group 1 received early tactile stimulation and Group 2 was a control group. The patients were assessed prospectively for 6 months for recovery of functional sensibility. Tactile stimulation in Group 1 was provided from 3 weeks after nerve repair with a specially designed rotating tactile stimulator and a pocket tactile stimulator. Constant two-point discrimination, moving two-point discrimination, and cutaneous pressure threshold were measured and sensibility was graded with the Medical Research Council (UK) sensibility grading. At 6 months, 68.8% of patients in Group 1 had a Medical Research Council grading of S3+ or S4 sensibility compared with 36 % in Group 2. With this prospective randomized study, the value of sensory reeducation in improving sensibility after digital nerve injury was confirmed. Starting tactile stimulation from the early postoperative period is recommended; however, use of the rotating tactile stimulator and pocket tactile stimulation need additional study.


CD4+ cell count responses to antiretroviral therapy are not impaired in HIV-infected individuals with tuberculosis co-infection.
To investigate whether HIV-infected individuals diagnosed with tuberculosis (HIV-TB) around the time of starting antiretroviral therapy (ART) have impaired CD4 cell responses to treatment. Analysis of a national cohort of HIV-infected adults, linked to the national TB surveillance system for England, Wales and Northern Ireland, including individuals starting ART from 2005 to 2009. We compared CD4 cell responses in HIV-infected individuals starting ART with a TB diagnosis ('HIV-TB cohort') with those not known to have TB ('TB-free cohort'). The TB-free cohort was frequency-matched to the HIV-TB cases for sex, age strata, baseline CD4 strata and ethnicity. Median change in CD4 cell count from baseline (ΔCD4) was calculated at 6-monthly intervals until 36 months. There were 593 and 1779 individuals in the HIV-TB and TB-free cohorts, respectively (median follow-up 3.8 years). In both cohorts, median age was 36 years, 49.2% were women and 74.9% were black-African. Median baseline CD4 at the start of treatment was similar in the HIV-TB and TB-free cohorts (74 vs. 80 cells/μl). Median ΔCD4 was similar in HIV-TB and TB-free cohorts at all time points [294 (inter-quartile range 198-424) cells/μl in HIV-TB cohort; 296 (inter-quartile range 196-431) cells/μl in TB-free cohort after 3 years of ART]. A higher proportion of the HIV-TB cohort than the TB-free cohort died during follow-up (4.2 vs. 2.2%; P = 0.01); 78.5% of all individuals who died had a baseline CD4 cell count below 100 cells/μl. Long-term CD4 cell recovery during ART appears similar in HIV-TB and TB-free patients. Significant mortality in both cohorts highlights the need for earlier HIV diagnosis and ART initiation.


Record rates of pressurized gas-flow in the great horsetail, Equisetum telmateia. Were Carboniferous Calamites similarly aerated?
Significant pressurized (convective) ventilation has been demonstrated in some flowering wetland plants, for example water-lilies and reeds, but not previously in nonflowering plants. Here we investigated convective flows in the great horsetail, Equisetum telmateia, and the possibility that convections aerated the massive rhizomes of the Calamites, extinct giant horsetails of the Carboniferous. Convection in E. telmateia was examined in relation to induction sites, anatomical pathways, relative humidity (RH), external wind-speed, diurnal effects, rhizome resistance and pressure-gradients. A mathematical model, incorporating Calamite aeration anatomy, was applied in assessing potentials for convective aeration. Individual shoots of E. telmateia generated extremely high rates of humidity-induced convection: < or = 120 cm(3) min(-1) (internal wind-velocity: 10 cm s(-1)) with rates proportional to branch numbers and 1/RH. Flows passed through branches, stem and rhizome via low-resistance lacunae (vallecular canals) and vented via stubble. Stomata supported internal pressures up to 800 Pa. Anatomically, E. telmateia resembles the Calamites and modelling predicted possible flows of 70 l min(-1) per Calamite tree. This is the first demonstration of significant convective flow in a nonflowering species, indicating that plant ventilation by a type of 'molecular gas-pump' may date back 350 million yr or more. Stomatal form and low-resistance pathways may facilitate high flow rates.


Sleep enhances nocturnal plasma ghrelin levels in healthy subjects.
Ghrelin, an endogenous ligand of the growth hormone secretagogue receptor, has been shown to promote slow-wave sleep (SWS, non-REM sleep stages 3 and 4). Plasma levels of ghrelin are dependent on food intake and increase in sleeping subjects during the early part of the night. It is unknown whether sleep itself affects ghrelin levels or whether circadian networks are involved. Therefore, we studied the effect of sleep deprivation on nocturnal ghrelin secretion. In healthy male volunteers, plasma levels of ghrelin, cortisol, and human growth hormone (hGH) were measured during two experimental sessions of 24 h each: once when the subjects were allowed to sleep between 2300 and 0700 and once when they were kept awake throughout the night. During sleep, ghrelin levels increased during the early part of the night and decreased in the morning. This nocturnal increase was blunted during sleep deprivation, and ghrelin levels increased only slightly until the early morning. Ghrelin secretion during the first hours of sleep correlated positively with peak hGH concentrations. We conclude that the nocturnal increase in ghrelin levels is more likely to be caused by sleep-associated processes than by circadian influences. During the first hours of sleep, ghrelin might promote sleep-associated hGH secretion and contribute to the promotion of SWS.


Post-irradiation malignant mesothelioma.
Approximately 30 patients with malignant mesothelioma following radiotherapy have been described. Population-based studies of this occurrence have not been reported. Patients with malignant mesothelioma of the pleura were collected. All of the patients had a prior cancer and had received radiotherapy to the region in which the malignant mesothelioma developed. Data from the National Cancer Institute's Surveillance, Epidemiology and End Results Program and the Connecticut Tumor Registry were evaluated for cases of malignant mesothelioma of the pleura occurring in patients with a previous cancer. The literature on post-irradiation malignant mesotheliomas was reviewed. Eight patients (4 men, 4 women) with malignant mesothelioma occurring in sites of radiotherapy for a prior tumor were identified. The mean age at diagnosis of mesothelioma was 45 years (range: 22-78 years), and the average interval between radiotherapy and the mesothelioma was 21 years (range: 11-29 years). Three of the patients had also received chemotherapy. Histologically, the mesotheliomas were epithelial in five cases, biphasic in one, and sarcomatous in one. One hundred forty-two patients were identified in the epidemiologic survey. The majority were men (89%), with a mean age for all patients of 68.5 years (range: 35-86 years) and a median latency between first cancer and mesothelioma of 4.3 years (range: 2 months-29.9 years). Mesotheliomas rarely develop as second malignant neoplasms. Within a large, population-based survey of patients with cancer, temporal patterns and demographic features of most second primary mesotheliomas were similar to asbestos-related tumors, although the late effects of cancer treatment might have contributed to the occurrence of cancer in some patients.


Cytokine-induced neutrophil chemoattractant release from hepatocytes is modulated by Kupffer cells.
To clarify the role of intercellular communication in the liver during accumulation of neutrophils, the release of cytokine-induced neutrophil chemoattractant (CINC) (interleukin-8 [IL-8] related protein in rodents) by hepatocytes was investigated in the presence of Kupffer cell-conditioned medium in vitro. Kupffer cells were prepared by perfusion of rat liver with collagenase followed by centrifugation on a metrizamide gradient and were cultured in the presence or absence of lipopolysacharide (LPS). The conditioned medium was collected after 24 hours, and rat hepatocytes were cultured in the presence or absence of Kupffer cell-conditioned medium. An amount of CINC in the culture supernatant was measured by western blotting analysis and enzyme-linked immunosorbent assay (ELISA), and expression of its messenger RNA (mRNA) was assessed by the polymerase chain reaction. LPS-stimulated Kupffer cell-conditioned medium enhanced an expression of CINC mRNA in hepatocytes and increased the production of CINC by hepatocytes. Enhanced production of CINC was not shown when the Kupffer cell-conditioned medium was pretreated with heat (56 degrees C, 30 minutes). The production of CINC by hepatocytes in the presence of the LPS-stimulated Kupffer cell-conditioned medium was reduced by an antibody against interleukin 1 beta (IL-1 beta), but not by antibodies against tumor necrosis factor alpha (TNF-alpha) or LPS. These results suggest that production of CINC by hepatocytes could be regulated by IL-1 beta released from Kupffer cells, leading to neutrophil accumulation during liver injury, because this protein is a strong chemoattractant for neutrophils.


Human Amniotic Membrane-Derived Mesenchymal and Epithelial Cells Exert Different Effects on Monocyte-Derived Dendritic Cell Differentiation and Function.
We previously demonstrated that mesenchymal cells from human amniotic membrane (hAMTCs) inhibit the generation and maturation of monocyte-derived dendritic cells (DCs) in vitro. Considering the crucial role of DCs in the immune response and that epithelial cells of the human amniotic membrane (hAECs) share some of the immunoregulatory properties of hAMTCs, we investigated whether hAECs also modulate monocyte-derived DCs. We compared hAECs with hAMTCs in a cell-to-cell contact setting and their secreted factors in modulating DC differentiation and function. First, we demonstrated that primary and expanded hAMTCs strongly inhibited the differentiation of DCs and induced a shift toward M2-like macrophages. This was observed when hAMTCs were cultured in contact (hAMTC-DC(cont)) or in Transwells (hAMTC-DC(tw)) with monocytes and even when medium conditioned by hAMTCs was used instead of hAMTCs. hAECs also prevented DC development, but to a lesser extent than hAMTCs. hAECs were more effective when cultured in contact with monocytes (hAEC-DC(cont)) rather than in Transwells (hAEC-DC(tw)). The modulatory capacity of hAECs changed during passaging unlike the hAMSCs. The ability to stimulate CD4(+) and CD8(+) T-cell proliferation was almost completely abolished by hAMTC-DC(cont), whereas hAMTC-DC(tw) and hAEC-DC(cont) displayed only a reduced ability to stimulate CD8(+) T cells. Furthermore, monocytes cocultured with hAMTCs and hAECs showed some similarities, but also differences in cytokine/chemokine secretion. Similarities were observed in the inhibition of IL-12p70 and TNF-α and the increase in IL-10 in supernatants taken from monocyte-DCs cocultured with hAMTCs and hAECs in contact and Transwell settings. The inflammatory factors IL-8, CXCL9, and MIP-1α were significantly lower in hAMTC-DC(cont), hAMTC-DC(tw), and hAEC-DC(cont) conditions. In contrast, only hAMTCs (in both contact and Transwell conditions) were able to significantly increase IL-1β and CCL2. Altogether, we demonstrated that hAMTCs and hAECs affect DC differentiation, but that hAMTCs exerted a stronger inhibitory effect, abolished T-cell proliferation, and also induced more changes in cytokine/chemokine production.


Efficient octopine Ti plasmid-derived vectors for Agrobacterium-mediated gene transfer to plants.
A two-component cloning system to transfer foreign DNA into plants was derived from the octopine Ti plasmid pTiB6S3. pGV2260 is a non-oncogenic Ti plasmid from which the T-region is deleted and substituted by pBR322. pGV831 is a streptomycin-resistant pBR325 derivative that contains a kanamycin resistance marker gene for plant cells and a site for cloning foreign genes between the 25-bp border sequences of the octopine T-region. Conjugative transfer of pGV831 derivatives to Agrobacterium and cointegration by homologous recombination between the pBR322 sequences present on pGV831 and pGV2260, can be obtained in a single step. Strains carrying the resulting cointegrated plasmids transfer and integrate T-DNA into the genome of tobacco protoplasts, and transformed tobacco calli are readily selected as resistant to kanamycin. Intact plants containing the entire DNA region between the T-DNA borders have been regenerated from such clones. In view of these properties we present pGV831 and its derivatives as vectors for efficient integration of foreign genes into plants.


Herpes encephalitis : a stroke mimicker.
Background: HSV-1 encephalitis (HSVE) usually presents with fever, altered mental status or focal seizures. Aphasia can also be a presenting symptom of HSVE but rarely occurs as the primary symptom. We present a case where aphasia was the primary presenting symptom of HSVE. Case: A 72-year-old physician with a history of hyperlipidemia and obstructive sleep apnea presented to the emergency room with sudden onset of speech difficulty lasting an hour. He did not have a fever, photophobia, neck stiffness, weakness, or numbness. The patient was brought in by the family within an hour to the emergency department. On exam, the only neurological deficit that was found was the use of inappropriate words in sentences and inability to name certain objects. He was diagnosed with an embolic stroke and received tPA. MRI brain that was done 24-hour post tPA showed an increased FLAIR and T2 signal hyperintensity within the medial left temporal lobe with slight effacement of the cysts sulci which was concerning for encephalitis. This was later confirmed by serology. The patient was started on IV Acyclovir and recovered fully after 3 weeks of acute neuro rehabilitation. Conclusion: Aphasia primarily is an unusual presentation of HSVE. It should be considered as one of the possibilities in patients presenting with features suggestive of a stroke involving the language areas of the brain.


Brief Report: Transgender Women and Preexposure Prophylaxis Care: High Preexposure Prophylaxis Adherence in a Real-World Health Care Setting in New York City.
Transgender women and trans feminine individuals (TGW/TFI) are a high priority population for the provision of HIV preexposure prophylaxis (PrEP) care within the United States, but there is limited research that focuses specifically on PrEP adherence within this population. Observational study of patients prescribed PrEP at a community-based health center. We enrolled 100 TGW/TFI PrEP patients at a community health center during clinic visits. Adherence data were collected at 3 time points, using self-report surveys, patient interviews, and urine assays measuring tenofovir. Data were summarized descriptively. The sample was diverse in age, race/ethnicity, and socioeconomic characteristics. Participants demonstrated strong PrEP adherence; at least 80% of the sample reported 90% or greater adherence at each time point. Concordance between self-report and urine assay was high. Among patients who reported taking PrEP within the past 48 hours, 82%-92% had detectable urine tenofovir. However, many patients reported PrEP stop periods of 4 or more days (28%-39% per time point). Our data highlight TGW/TFI's capacity to adhere to daily PrEP and sustain PrEP use over time. The concordance between patient self-report and urine TFV levels suggest that providers can trust patient reports of PrEP adherence behavior and support the use of adherence conversations in clinical settings, without the need for point of care biological monitoring. Findings also underscore the importance of continued attention to drivers of PrEP stops at the patient, clinic, and systems levels and the development of strategies that support sustained PrEP use.


A post-marketing study on interferon beta 1b and 1a treatment in relapsing-remitting multiple sclerosis: different response in drop-outs and treated patients.
Interferon beta 1b (Betaferon) and 1a (Avonex) were licensed in Italy for treating relapsing-remitting multiple sclerosis in February 1996 and August 1997, respectively. To evaluate the effectiveness of these agents on the basis of clinical experience in northern Italian multiple sclerosis centres. Clinical data on patients with relapsing-remitting multiple sclerosis were collected on an appropriate form from 65 centres in northern Italy. Intention to treat analysis was not possible, so patients who discontinued treatment (drop-outs) and who continued treatment (treated) were analysed separately. The main outcome measures were annual relapse frequency, number of relapse-free patients, mean change in extended disability status scale score (EDSS), and number of patients who worsened. 1481 patients were included; 834 were treated with Betaferon and 647 with Avonex for mean periods of 21.4 and 12.0 months, respectively. Basal EDSS was 2.37 and 2.17, respectively, and relapse frequency was 1.62 and 1.45. The annual relapse rate decreased by more than 60% with Betaferon and 55% with Avonex. The proportions of relapse-free, improved, and worsened patients were similar in the two groups. More patients interrupted treatment with Betaferon (41.1%) than with Avonex (15.3%); such patients showed more active disease at baseline and during treatment. The incidence of side effects was higher in Betaferon treated patients. The effectiveness of Betaferon and Avonex is confirmed. There was a more marked effect than expected from the experimental trial results. This might reflect differences in inclusion criteria, or, more likely, loss of drop-outs, favouring selective retention of responders.