NF-IL6-mediated transcriptional activation of the long terminal repeat of the human immunodeficiency virus type 1.
An upstream control region in the long terminal repeat (LTR) of human immunodeficiency virus type 1 (HIV-1) includes a potential negative regulatory element (NRE1). Cotransfecting multimers of a sequence spanning this element with an LTR-CAT construct produced an increase in chloramphenicol acetyltransferase (CAT) activity in Jurkat and HepG2 cells, providing further evidence and support for the existence of an NRE. In screening experiments aimed at identifying those factors that regulate HIV-1 transcription through interactions with the NRE1 region, we isolated a cDNA for NF-IL6. Previous studies have shown that NF-IL6 is a key nuclear factor that activates gene expression in response to interleukin 6. By methylation interference analysis, we have localized the NF-IL6 binding site within the NRE1 region and found that it overlaps an E box that has previously been implicated as the binding element for a negative regulator of HIV-1 expression. Through a database search, we identified an additional consensus binding sequence for NF-IL6 in the LTR of many HIV-1 variants and found that over this sequence, purified NF-IL6 can produce an extended footprint that overlaps one of the binding sites for NF-kappa B. A product of the nf-il6 gene activated transcription from several LTR-CAT constructs in transient transfection assays. Thus, NF-IL6 could play a central role in the control of HIV-1 gene expression and this protein might be a key mediator in signaling pathways where HIV-1 is activated by interleukin 6.


Changes in surface area of intact guard cells are correlated with membrane internalization.
Guard cells must maintain the integrity of the plasma membrane as they undergo large, rapid changes in volume. It has been assumed that changes in volume are accompanied by changes in surface area, but mechanisms for regulating plasma membrane surface area have not been identified in intact guard cells, and the extent to which surface area of the guard cells changes with volume has never been determined. The alternative hypothesis-that surface area remains approximately constant because of changes in shape-has not been investigated. To address these questions, we determined surface area for intact guard cells of Vicia faba as they underwent changes in volume in response to changes in the external osmotic potential. We also estimated membrane internalization for these cells. Epidermal peels were subjected to external solutions of varying osmotic potential to shrink and swell the guard cells. A membrane-specific fluorescent dye was used to identify the plasma membrane, and confocal microscopy was used to acquire a series of optical paradermal sections of the guard cell pair at each osmotic potential. Solid digital objects representing the guard cells were created from the membrane outlines identified in these paradermal sections, and surface area, volume, and various linear dimensions were determined for these solid objects. Surface area decreased by as much as 40% when external osmotic potential was increased from 0 to 1.5 MPa, and surface area varied linearly with volume. Membrane internalization was approximated by determining the amount of the fluorescence in the cell's interior. This value was shown to increase approximately linearly with decreases in the cell's surface area. The changes in surface area, volume, and membrane internalization were reversible when the guard cells were returned to a buffer solution with an osmotic potential of approximately zero. The data show that intact guard cells undergo changes in surface area that are too large to be accommodated by plasma membrane stretching and shrinkage and suggest that membrane is reversibly internalized to maintain cell integrity.


Imipramine disposition in users of oral contraceptive steroids.
Ten women on long-term, low-dose estrogen oral contraceptive steroids (OCS) and eight age-matched drug-free female controls received an intravenous infusion of 12.5 mg imipramine. Eleven (six OCS users and five controls) also took a 50-mg oral dose of imipramine on another occasion. After intravenous injection, volume of distribution was much the same in the OCS and control groups and clearance was of the same order (899 and 975 ml/min). Elimination t1/2 was prolonged in OCS users after intravenous doses (17.8 vs 25.5 hr) but did not change after oral doses (18.4 vs 19.1 hr). Imipramine plasma protein binding was of the same order in both groups. Absolute bioavailability increased in OCS users (from 27.1% vs 44.1%), which resulted in a trend toward decreased apparent oral clearance (from 4649 vs 2322 ml/min). Women who used OCS regularly show little change in imipramine kinetics after intravenous dosing. After oral dosing absolute systemic bioavailability increased, resulting in decreased apparent oral clearance in the absence of any change in oral elimination t1/2. Imipramine (with high first-pass hepatic extraction) is nonrestrictively cleared, with drug elimination predominantly a function of hepatic blood flow. These data are consistent with OCS inhibition of hepatic imipramine oxidation with no alteration in hepatic blood flow resulting from chronic OCS use.


Immunohistochemical Analysis of ATRX, IDH1 and p53 in Glioblastoma and Their Correlations with Patient Survival.
Glioblastoma (GBM) can be classified into molecular subgroups, on the basis of biomarker expression. Here, we classified our cohort of 163 adult GBMs into molecular subgroups according to the expression of proteins encoded by genes of alpha thalassemia/mental retardation syndrome X-linked (ATRX), isocitrate dehydrogenase (IDH) and TP53. We focused on the survival rate of molecular subgroups, depending on each and various combination of these biomarkers. ATRX, IDH1 and p53 protein expression were evaluated immunohistochemically and Kaplan-Meier analysis were carried out in each group. A total of 15.3% of enrolled GBMs demonstrated loss of ATRX expression (ATRX-), 10.4% expressed an aberrant IDH1 R132H protein (IDH1+), and 48.4% exhibited p53 overexpression (p53+). Survival differences were statistically significant when single protein expression or different combinations of expression of these proteins were analyzed. In conclusion, in the case of single protein expression, the patients with each IDH1+, or ATRX-, or p53- GBMs showed better survival than patients with counterparts protein expressed GBMs. In the case of double protein pairs, the patients with ATRX-/p53-, ATRX-/IDH1+, and IDH1+/p53- GBMs revealed better survival than the patients with GBMs with the remained pairs. In the case of triple protein combinations, the patients with ATRX-/p53-/IDH+ showed statistically significant survival gain than the patients with remained combination of proteins-expression status. Therefore, these three biomarkers, individually and as a combination, can stratify GBMs into prognostically relevant subgroups and have strong prognostic values in adult GBMs.


Sheep and goats raised in mixed flocks have diverse immune status around parturition.
Several physiological and metabolic changes take place in dairy ruminants around parturition (late pregnancy, parturition, and early lactation). Dairy species are genetically selected for their higher milk production compared with non-dairy species. This fact causes a constant stress that impairs the immune status of the animal, with consequences for its welfare and performance. In the present study, we assessed the immune status of high-yield dairy sheep and goats by quantifying IgG and IgM concentrations, as well as chitotriosidase (ChT) and complement system [total complement system (TC) and alternative complement pathway (AC)] activity in blood plasma around parturition. We also measured IgG and IgM concentrations and ChT activity in colostrum and milk during the first 40 d postpartum. The lowest blood IgG concentration was at parturition in both species. We detected no differences in blood IgG concentrations between species. Blood IgM concentrations were constant in both species throughout the study period. However, blood IgM concentrations were greater in sheep than in goats. Blood ChT activity was greater in goats than in sheep, and both species showed constant activity of this enzyme throughout the study period. We observed no differences in complement system (TC and AC) activity between sheep and goats. In addition, both TC and AC activity were constant in both species throughout the experiment. In general, IgG and IgM concentrations were greater in sheep colostrum than in goat colostrum, but these differences disappeared after d 4 (IgG) and d 3 (IgM) postpartum. In both species, the highest IgG and IgM concentrations were measured in colostrum, gradually decreasing during the first days postpartum. Chitotriosidase activity decreased in both species from colostrum to milk, although goats always showed greater ChT activity than sheep. Both sheep and goats seemed to be more susceptible to infectious diseases around parturition. As well, goats showed greater ChT activity in blood, colostrum, and milk than sheep. This fact may give these animals additional protection against parasite and fungal infections.


Control of virus diseases in soybeans.
Soybean, one of the world's most important sources of animal feed and vegetable oil, can be infected by numerous viruses. However, only a small number of the viruses that can potentially infect soybean are considered as major economic problems to soybean production. Therefore, we consider management options available to control diseases caused by eight viruses that cause, or have the potential to cause, significant economic loss to producers. We summarize management tactics in use and suggest direction for the future. Clearly, the most important tactic is disease resistance. Several resistance genes are available for three of the eight viruses discussed. Other options include use of virus-free seed and avoidance of alternative virus hosts when planting. Attempts at arthropod vector control have generally not provided consistent disease management. In the future, disease management will be considerably enhanced by knowledge of the interaction between soybean and viral proteins. Identification of genes required for soybean defense may represent key regulatory hubs that will enhance or broaden the spectrum of basal resistance to viruses. It may be possible to create new recessive or dominant negative alleles of host proteins that do not support viral functions but perform normal cellular function. The future approach to virus control based on gene editing or exploiting allelic diversity points to necessary research into soybean-virus interactions. This will help to generate the knowledge needed for rational design of durable resistance that will maximize global production.


Antimicrobial peptides modulate lung injury by altering the intestinal microbiota.
Mammalian mucosal barriers secrete antimicrobial peptides (AMPs) as critical, host-derived regulators of the microbiota. However, mechanisms that support microbiota homeostasis in response to inflammatory stimuli, such as supraphysiologic oxygen, remain unclear. We show that supraphysiologic oxygen exposure to neonatal mice, or direct exposure of intestinal organoids to supraphysiologic oxygen, suppresses the intestinal expression of AMPs and alters intestinal microbiota composition. Oral supplementation of the prototypical AMP lysozyme to hyperoxia-exposed neonatal mice reduced hyperoxia-induced alterations in their microbiota and was associated with decreased lung injury. Our results identify a gut-lung axis driven by intestinal AMP expression and mediated by the intestinal microbiota that is linked to lung injury in newborns. Together, these data support that intestinal AMPs modulate lung injury and repair. Video Abstract.


Psychological characteristics of children with Shwachman syndrome.
Twelve children and young adults with Shwachman syndrome were compared with their unaffected siblings and with controls suffering from cystic fibrosis in terms of intellectual ability, motor skills, and behaviour. There were highly significant differences in intelligence quotient between those with Shwachman syndrome and the other two groups. Four of the index subjects but none of the control subjects were below the normal range. The differences between groups on other tests of cognitive and motor skills were not significant, though those with Shwachman syndrome tended to have the lowest scores. There was no evidence that those with Shwachman syndrome had more behavioural difficulties than the control subjects. We suggest that the intellectual difficulties of patients with Shwachman syndrome may be of neurological rather than social origin and that they may originate before birth.


An atypical course of myasthenia gravis.
Myasthenia gravis (MG) is a chronic neuromuscular disease which leads to varying degrees of weakness in the skeletal muscles. Some of the symptoms of the disorder include weakness of the eye muscles, difficulty in swallowing and slurred speech. When only the muscles of the eyes are affected, the illness is termed ocular myasthenia, which is often characterized by abrupt onset of diplopia and ptosis of the eyelid. In most patients with ocular-onset MG, there is a progression to involvement of other muscle groups within the first two years (generalized myasthenia). In the case reported here, a 39-year-old male of Ecuadorian descent complained of difficulty seeing, double vision, dizziness, unsteady gait, difficulty maintaining balance and fatigue for the previous two days. Neurological examination was remarkable for total external ophthalmoplegia. There was no external bulbar muscle paralysis, motor weakness, muscle wasting, sensory deficits or sphincter dysfunction. His laboratory workup was significant for elevated acetylcholine receptor antibody. He was diagnosed with ocular MG after differential diagnoses were ruled out based on the onset and presentation of symptoms, the patient's age and a normal magnetic resonance imaging exam. No signs of generalized myasthenia were detected. His symptoms improved dramatically after treatment with Acetyl cholinesterase (AchE) inhibitors and steroids, regaining much of his ocular mobility and ability to walk without gait imbalance. At follow-up visits, the patient remained healthy with no evidence development of other myasthenic signs. This case is atypical since ocular MG does not normally occur in the absence of other myasthenic forms.


Anti-lipid-peroxidative principles from Tournefortia sarmentosa.
Using the inhibition of Cu(2+)-induced low-density-lipoprotein (LDL) peroxidation to direct fractionation, four new benzenoids, tournefolal (1), tournefolic acids A (2) and B (3), and B ethyl ester (4), together with salvianolic acid A (5), isosalvianolic acid C (6), lithospermic acid (7), salvianolic acid F (8), and rosmarinic acid (9), were isolated from the stems of Tournefortia sarmentosa. The structures of the new compounds 1-4 were elucidated on the basis of spectral and chemical methods. Furthermore, the anti-LDL-peroxidative activity of the isolated compounds was determined. All isolated compounds exhibited more potent activity than probucol except for salvianolic acid F (8).


Molecular subtypes of breast cancers detected in mammography screening and outside of screening.
The frequency and significance of gene expression profile-derived molecular subtypes of breast cancers found in mammography screening are unknown. We identified breast cancers diagnosed in women of any age living in defined geographic regions in Finland in 1991 to 1992 and collected clinical and pathologic data. Surrogates for the molecular subtypes were determined for 247 cancers found in organized mammography screening and 989 cancers detected outside of screening using immunohistochemistry or in situ hybridization. Molecular subtypes were defined as luminal A [estrogen receptor (ER) positive and/or progesterone receptor (PR) positive, HER2-], luminal B (ER+ and/or PR+, HER2+), basal-like (ER-, PR-, HER2-, cytokeratin 5+, and/or HER1+), HER2+/ER- (ER-, PR-, and HER2+), and unclassified. The median follow-up time was 9.4 years. The luminal type A was common (73.7%) and the HER2+/ER- type is rare (5.7%) in screen-detected cancer, and only 16% were HER2 positive. Women with cancer diagnosed in screening at ages 50 to 69 years had similar molecular subtype distribution as women whose cancer was found outside of screening at age >69 years. In a multivariate model, cancer detection at screening independently predicted favorable distant disease-free survival when the molecular subtype was included as a covariate in addition to age, histologic grade, and cancer size. Women with small (pT(1)N(0)M(0)) HER2-positive cancer had similar outcome regardless of the method of detection. Molecular subtype distribution of screen-detected breast cancer differs from that of cancers found outside of screening and accounts in part for the better outcome of screen-detected cancer.


PET Cell Tracking Using 18F-FLT is Not Limited by Local Reuptake of Free Radiotracer.
Assessing the retention of cell therapies following implantation is vital and often achieved by labelling cells with 2'-[18F]-fluoro-2'-deoxy-D-glucose (18F-FDG). However, this approach is limited by local retention of cell-effluxed radiotracer. Here, in a preclinical model of critical limb ischemia, we assessed a novel method of cell tracking using 3'-deoxy-3'-L-[18F]-fluorothymidine (18F-FLT); a clinically available radiotracer which we hypothesise will result in minimal local radiotracer reuptake and allow a more accurate estimation of cell retention. Human endothelial cells (HUVECs) were incubated with 18F-FDG or 18F-FLT and cell characteristics were evaluated. Dynamic positron emission tomography (PET) images were acquired post-injection of free 18F-FDG/18F-FLT or 18F-FDG/18F-FLT-labelled HUVECs, following the surgical induction of mouse hind-limb ischemia. In vitro, radiotracer incorporation and efflux was similar with no effect on cell viability, function or proliferation under optimised conditions (5 MBq/mL, 60 min). Injection of free radiotracer demonstrated a faster clearance of 18F-FLT from the injection site vs. 18F-FDG (p ≤ 0.001), indicating local cellular uptake. Using 18F-FLT-labelling, estimation of HUVEC retention within the engraftment site 4 hr post-administration was 24.5 ± 3.2%. PET cell tracking using 18F-FLT labelling is an improved approach vs. 18F-FDG as it is not susceptible to local host cell reuptake, resulting in a more accurate estimation of cell retention.


Role of medial amygdala in controlling hemodynamics via GABA(A) receptor in anesthetized rats.
Experiments were conducted to determine whether the medial amygdala could control hemodynamics through the GABA receptor in anesthetized rats. Bicuculline (10-100pmol), a GABA(A) antagonist, injected into the anterior basomedial amygdala (medial amygdala) produced dose-related increases in blood pressure (BP) and heart rate (HR). Following microinjection of a 100pmol dose, plasma norepinephrine and epinephrine also rose significantly. In a group that had undergone bilateral adrenalectomy, the same dose of bicuculline into the same region of the amygdala caused similar increases in BP and HR to those produced in normal rats. Pretreatment with an intraperitoneal injection of hexamethonium prevented these responses. When bicuculline was given intravenously (100pmol) it failed to effect these cardiovascular changes. Bicuculline-induced hypertension and tachycardia were inhibited by microinjection of muscimol, a GABA agonist, into the anterior basomedial amygdala. The results thus demonstrate that the influence of bicuculline on BP and HR is through action on the anterior basomedial amygdala and direct sympathetic outflow to heart and vascular smooth muscle. There is tonic GABAergic inhibition of sympathetic outflow in the medial amygdala that plays a part in regulating hemodynamics in the limbic system.


[Expression of inducible nitric oxide synthase was regulated by activating nuclear factor kappaB in mouse macrophages stimulated with ureaplasma urealyticum lipid-associated membrane proteins].
The aim was to investigate the molecular mechanisms responsible for the inducible nitric oxide synthase (iNOS) gene expression stimulated by lipid associated membrane proteins (LAMPs) of Ureaplasma urealyticum (Uu). Mouse macrophages were stimulated by Ureaplasma urealyticum LAMPs to analyze the production of nitric oxide (NO) and the expression of iNOS detected by RT-PCR and Western blot. The activation of NF-kappaB was examined in mouse macrophages treated with LAMPs by indirect immunofluorescence (IFA), immunocytochemistry and Western blot. The effects of pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kappaB and of cycloheximide (CHX), a protein synthase inhibitor, on the expression of iNOS and on the activation of NF-kappaB were also investigated in mouse macrophages treated with LAMPs. Results showed Ureaplasma urealyticum LAMPs stimulated mouse macrophages to express iNOS and thus produce NO in dose- and time-dependent manners by activating nuclear factor kappaB. The activation of NF-kappaB and the expression of iNOS were inhibited by LAMPs combination with PDTC or CHX. In conclusion, these findings suggested Ureaplasma urealyticum may be an important pathogenic factor due to the ability of LAMPs to stimulate the expression of iNOS, which is probably medicated by the activation of NF-kappaB.


Novel Canthin-6-one Derivatives: Design, Synthesis, and Their Antiproliferative Activities via Inducing Apoptosis, Deoxyribonucleic Acid Damage, and Ferroptosis.
A series of novel canthin-6-one (CO) derivatives (8a-l) were designed and synthesized by introducing different amide side chains at the C-2 position, and their water solubility, antiproliferative activity, and preliminary mechanism were investigated. Most compounds displayed high cytotoxicity exhibiting low-micromolar IC50 values against four human cancer cell lines, especially HT29 cells. Meanwhile, the water solubility of active CO derivatives was significantly improved. Among these compounds, compound 8h with the N-methyl piperazine group exhibiting the highest antiproliferative capability with an IC50 value of 1.0 μM against HT29 cells, which was 8.6-fold lower than that of CO. Furthermore, 8h could upregulate the levels of reactive oxygen species, leading to mitochondrial damage. In addition, 8h could promote cell apoptosis and DNA damage by regulating the expression of apoptosis-associated proteins (Bcl-2 and cleaved-caspase 3) and the DNA damage-associated protein (H2AX). Most importantly, 8h also exerted ferroptosis by reducing the GSH level and GPX4 expression as well as increasing the lipid peroxidation level. Thus, the novel CO derivative 8h with N-methylpiperazine represents a promising anticancer candidate and warrants a more intensive study.


Humoral and cellular immune memory to four COVID-19 vaccines.
Multiple COVID-19 vaccines, representing diverse vaccine platforms, successfully protect against symptomatic COVID-19 cases and deaths. Head-to-head comparisons of T cell, B cell, and antibody responses to diverse vaccines in humans are likely to be informative for understanding protective immunity against COVID-19, with particular interest in immune memory. Here, SARS-CoV-2-spike-specific immune responses to Moderna mRNA-1273, Pfizer/BioNTech BNT162b2, Janssen Ad26.COV2.S and Novavax NVX-CoV2373 were examined longitudinally for 6 months. 100% of individuals made memory CD4 + T cells, with cTfh and CD4-CTL highly represented after mRNA or NVX-CoV2373 vaccination. mRNA vaccines and Ad26.COV2.S induced comparable CD8 + T cell frequencies, though memory CD8 + T cells were only detectable in 60-67% of subjects at 6 months. Ad26.COV2.S was not the strongest immunogen by any measurement, though the Ad26.COV2.S T cell, B cell, and antibody responses were relatively stable over 6 months. A differentiating feature of Ad26.COV2.S immunization was a high frequency of CXCR3 + memory B cells. mRNA vaccinees had substantial declines in neutralizing antibodies, while memory T cells and B cells were comparatively stable over 6 months. These results of these detailed immunological evaluations may also be relevant for vaccine design insights against other pathogens.


PERSPECTIVE: INDIRECT MATE CHOICE, COMPETITION FOR MATES, AND COEVOLUTION OF THE SEXES.
When Darwin first proposed the possibility of sexual selection, he identified two mechanisms, male competition for mates and female choice of mates. Extending this classification, we distinguish two forms of mate choice, direct and indirect. This distinction clarifies the relationship between Darwin's two mechanisms and, furthermore, indicates that the potential scope for sexual selection is much wider than thus far realized. Direct mate choice, the focus of most research on sexual selection in recent decades, requires discrimination between attributes of individuals of the opposite sex. Indirect mate choice includes all other behavior or morphology that restricts an individual's set of potential mates. Possibilities for indirect mate choice include advertisement of fertility or copulation, evasive behavior, aggregation or synchronization with other individuals of the same sex, and preferences for mating in particular locations. In each of these cases, indirect mate choice sets the conditions for competition among individuals of the opposite sex and increases the chances of mating with a successful competitor. Like direct mate choice, indirect mate choice produces assortative mating. As a consequence, the genetic correlation between alleles affecting indirect choice and those affecting success in competition for mates can produce self-accelerating evolution of these complementary features of the sexes. The broad possibilities for indirect mate choice indicate that sexual selection has more pervasive influences on the coevolution of male and female characteristics than previously realized.


Plasmodium falciparum: in vitro chloroquine susceptibility and allele-specific PCR detection of Pfmdr1 Asn86Tyr polymorphism in Lambarene, Gabon.
Plasmodium falciparum resistance to chloroquine has been described in many parts of the world particularly in Africa where malaria is endemic. High levels of chloroquine resistance in our study area, Lambarene-Gabon, has led to the use of an alternative regimen for treatment and prevention of P. falciparum infection. In this study, we examined the in vitro chloroquine sensitivity of 15 isolates from this area and assessed the prevalence of a putative chloroquine resistance associated Pfmdr1 polymorphism (Asn86Tyr) using a novel allele-specific polymerase chain reaction (PCR). Only 4 of the isolates examined were chloroquine sensitive. The allele-specific PCR shows that all 15 isolates carried the variant (86Tyr) codon. Eleven of these were resistant to chloroquine suggesting a 73% agreement between chloroquine resistance phenotype and the point mutation. This molecular marker was examined in a further 73 Gabonese isolates, where 58 (79.5%) showed 86Tyr and 15 (20.5%) showed 86Asn. In all, 4 (4.5%) of the 88 isolates assessed carry both mutant and wild-type codons, suggesting mixed parasite populations. The incomplete agreement found between chloroquine resistance phenotype and Pfmdr1 (86Tyr) polymorphism would support the view that other genetic factors as well as Pfmdr1 may be involved in chloroquine resistance. While our results suggest a high prevalence of 86Tyr polymorphism in Lambarene, the Asp1246Tyr polymorphism (a point mutation which to date has only been associated with South American P. falciparum) seems to be absent in our study area.


A squarate-pillared titanium oxide quantum sieve towards practical hydrogen isotope separation.
Separating deuterium from hydrogen isotope mixtures is of vital importance to develop nuclear energy industry, as well as other isotope-related advanced technologies. As one of the most promising alternatives to conventional techniques for deuterium purification, kinetic quantum sieving using porous materials has shown a great potential to address this challenging objective. From the knowledge gained in this field; it becomes clear that a quantum sieve encompassing a wide range of practical features in addition to its separation performance is highly demanded to approach the industrial level. Here, the rational design of an ultra-microporous squarate pillared titanium oxide hybrid framework has been achieved, of which we report the comprehensive assessment towards practical deuterium separation. The material not only displays a good performance combining high selectivity and volumetric uptake, reversible adsorption-desorption cycles, and facile regeneration in adsorptive sieving of deuterium, but also features a cost-effective green scalable synthesis using chemical feedstock, and a good stability (thermal, chemical, mechanical and radiolytic) under various working conditions. Our findings provide an overall assessment of the material for hydrogen isotope purification and the results represent a step forward towards next generation practical materials for quantum sieving of important gas isotopes.


Storage-Dependent Generation of Potent Anti-ZIKV Activity in Human Breast Milk.
Zika virus (ZIKV) causes congenital neurologic birth defects, notably microcephaly, and has been associated with other serious complications in adults. The virus has been detected in human breast milk and possible transmissions via breastfeeding have been reported. Breast milk is rich in nutrients and bio-active substances that might directly affect viral infectivity. Thus, here, we analyzed the effect of human breast milk on ZIKV infection. We observed that fresh human breast milk had no effect on ZIKV, but found that upon storage, milk effectively suppressed infection. The antiviral activity is present in the fat-containing cream fraction of milk and results in the destruction of the structural integrity of viral particles, thereby abrogating infectivity. The release of the factor is time dependent but varies with donors and incubation temperatures. The viral titer of milk that was spiked with ZIKV decreased considerably upon storage at 37 °C for 8 h, was lost entirely after 2 days of 4 °C storage, but was not affected at -20 °C. This suggests that cold storage of milk inactivates ZIKV and that the antiviral factor in milk may also be generated upon breastfeeding and limit this transmission route of ZIKV.


The Processing of Surgical Specimens With Forensic Evidence: Lessons Learned From the Boston Marathon Bombings.
Following the Boston Marathon bombings in April 2013, pathology departments at hospitals across Boston, Massachusetts received numerous amputated limbs, as well as other surgical specimens from trauma surgeries. In the absence of clear guidelines, each department faced uncertainties in performing gross examination of these specimens. To develop a protocol for processing surgical specimens with forensic evidence. We collaborated with representatives who knew the practices at 3 major Boston hospitals, the Office of the Chief Medical Examiner of Massachusetts, and a senior team leader for the evidence response team for the Boston, Massachusetts division of the US Federal Bureau of Investigation to construct a protocol for processing specimens with forensic evidence. A simple and robust protocol approved by experts in forensic evidence collection was developed. Important points in this protocol include (1) assigning the task of processing the specimens to one individual or one team of individuals, (2) photographing all specimens before and after washing, (3) obtaining a radiograph of each specimen, and (4) identifying a secure area to store forensic evidence. When acts of terror occur, protocols provide order and clarification to the processing of surgical specimens. We propose a protocol that provides guidance for pathology departments across the country to handle trauma-related surgical specimens with forensic evidence in an efficient manner to allow optimal patient care and a secure way of gathering forensic evidence.


Overview of the MODARIA programme (and comments on implications for future programmes of work).
The International Atomic Energy Agency (IAEA) has organised programmes on the development, comparison and testing of environmental assessment models and approaches for estimating the radiation exposure of humans and wildlife since the 1980s. The latest of these programmes was called MODARIA (Modelling and Data for Radiological Impact Assessment) and was run in two phases from 2012 to 2015 (MODARIA I) and 2016 to 2019 (MODARIA II). Both phases of the MODARIA programme had the overall objective to improve capabilities in the field of environmental transfer of radionuclides and public and non-human biota exposures assessment, by means of acquisition of improved data for model testing and comparison, reaching consensus on modelling philosophies, approaches and parameter values and building an international forum for the exchange of information. This paper provides an overview of the work undertaken during both phases of the MODARIA programme and its outputs. The overall aims and objectives of a new programme to follow on from MODARIA are described.


A base-pairing interaction between U2 and U6 small nuclear RNAs occurs in > 150S complexes in HeLa cell extracts: implications for the spliceosome assembly pathway.
In mammalian cells, base pairing between the U2 and U6 small nuclear RNAs is required during pre-RNA splicing. We show by psoralen crosslinking of HeLa nuclear extract that U2.U6 base pairing occurs within abundant ribonucleoprotein complexes that sediment at > 150 S in glycerol gradients. All of the spliceosomal RNAs (U1, U2, U4, U5, and U6) cosediment with these large complexes, suggesting that they may be related to small nuclear RNA-containing structures called speckles/coiled bodies or snurposomes, which have been visualized in mammalian or amphibian nuclei, respectively. In contrast to nuclear extract, S100 extract, which is splicing-defective and lacks the > 150S complexes, does not contain base-paired U2.U6. However, U2.U6 base pairs form in S100 extract that has been made splicing-competent by supplementation with Ser/Arg-rich (SR) proteins, ATP, and an adenovirus splicing substrate. During splicing in supplemented S100 extract, U2.U6 base pairing precedes the appearance of splicing intermediates and occurs initially in an approximately 60S spliceosome complex but also in progressively larger (100-300 S) complexes. Possible functional relationships between the 60S spliceosome and the > 150S complexes are discussed.


[Spinal cord compression caused by Cladosporium trichoides. An anatomo-clinical case report].
The authors report a case concerning a 25-year-old man who presented a cervical cord compression. The pathologic examination of the mass disclosed a chronic granulomatous meningitis with dense fibrosis. Septate and pigmented hyphae of Cladosporium trichoides were present among the inflammatory infiltrate. A review of the literature concerning the involvement of the central nervous system by this agent is performed. This is the first case in which the disease started as a compressive cord syndrome.


Enterolactone Reduces Telomerase Activity and The Level of Its Catalytic Subunit in Breast Cancer Cells.
There is a positive correlation between higher serum phytoestrogen concentrations and lower risk of breast cancer. The activation of telomerase is crucial for the growth of cancer cells; therefore, the aim of this study was to examine the effects of enterolactone (ENL) and enterodiol (END) on this enzyme. In this experimental study, we performed the viability assay to determine the effects of different concentrations of ENL and END on cell viability, and the effective concentrations of these two compounds on cell growth. We used western blot analysis to evaluate human telomerase reverse transcriptase catalytic subunit (hTERT) expression and polymerase chain reaction (PCR)-ELISA based on the telomeric repeat amplification protocol (TRAP) assay for telomerase activity. Both ENL and END, at 100 μM concentrations, significantly (P<0.05) reduced cell viability. However, only the 100 μM concentration of ENL significantly (P<0.05) decreased hTERT protein levels and telomerase activity. Lower concentrations of ENL did not have any significant effects on telomerase activity and hTERT protein levels. High concentration of ENL decreased the viability of MCF-7 breast cancer cells and inhibited the expression and activity of telomerase in these cells. Although END could reduce breast cancer cell viability, it did not have any effect on telomerase expression and activity.


In vivo and in vitro modulation of carp (Cyprinus carpio L.) phagocyte oxidative burst activity by gallium.
Since gallium is a metal ion used in semiconductor industry, the toxicological effects were previously evaluated in mammals but the ecotoxicological impacts remain unknown. In term of ecotoxicological risk assessment, the median lethal concentration (LC50 for 96 h of gallium to carp (Cyprinus carpio L.) and the oxidative response of carp phagocytes after the fish were exposed to sublethal levels of gallium were determined. The LC50 of gallium on C. carpio at 96 h was estimated as 96.25 +/- 14.3 mg/L. To determine the effect in vivo of gallium on the phagocyte response, fish were exposed for 96 h to 5 or 50 mg Ga(3+)/L. Carp maintained for 48 or 96 h in water containing 50 mg/L gallium had a significant fall in phagocyte oxidative burst activity in comparison with controls, as well as decreased leukocyte number in blood and increased cytotoxicity. To determine the effect in vitro of gallium on the phagocyte response, isolated phagocytes were exposed for 5 or 15 min to 50 nM, 500 nM, or 5, 50, 100, or 200 microM of Ga(3+). The oxidative burst was increased after in vitro incubation of phagocytes with 50 or 500 nM gallium for 15 min or with 500 nM gallium for 5 min. Moreover, for 50, 100, or 200 microM gallium, the oxidative burst activity of carp phagocytes was significantly decreased. Results indicate that the lethal toxicity of gallium for carp of gallium is not as high as for other metal ions. However, gallium was immunosuppressive for carp at the highest concentrations used (from 50 microM) in vivo and in vitro. At low concentrations, it could be an immunostimulant as observed in vitro.


Nitrogen-Doped Graphene via In-situ Alternating Voltage Electrochemical Exfoliation for Supercapacitor Application.
Doping heteroatom, an effective way to enhance the electrochemical performances of graphene, has received wide attention, especially related to nitrogen. Alternating voltage electrochemical exfoliation, as a low cost and green electrochemical approach, has been developed to construct in-situ N-doped graphene (N-Gh) material. The N-Gh presents a much higher capacity than that of pure graphene prepared via the same method, which might be attributed to the introduction of nitrogen, which has much more effects and a disordered structure. As-prepared N-Gh exhibits a low O/C ratio that is helpful in maintaining high electrical conductivity. And the effects and disorder structure are also conductive to reduce the overlaps of graphene layers. A symmetric supercapacitor assembled with N-Gh electrodes displays a satisfactory rate behavior and long cycling stability (92.3% retention after 5,000 cycles).


The secret hidden in dust: Assessing the potential to use biological and chemical properties of the airborne fraction of soil for provenance assignment and forensic casework.
The airborne fraction of soil (dust) is both ubiquitous in nature and contains localised biological and chemical signatures, making it a potential medium for forensic intelligence. Metabarcoding of dust can yield biological communities unique to the site of interest, similarly, geochemical analyses can uncover elements and minerals within dust that can be matched to a geographic location. Combining these analyses presents multiple lines of evidence as to the origin of dust collected from items of interest. In this work, we investigated whether bacterial and fungal communities in dust change through time and whether they are comparable to soil samples of the same site. We integrated dust metabarcoding into a framework amenable to forensic casework, (i.e., using calibrated log-likelihood ratios) to predict the origin of dust samples using models constructed from both dust samples and soil samples from the same site. Furthermore, we tested whether both metabarcoding and geochemical/mineralogical analyses could be conducted on a single swabbed sample, for situations where sampling is limited. We found both analyses could generate results from a single swabbed sample and found biological and chemical signatures unique to sites. However, we did find significant variation within sites, where this did not always correlate with time but was a random effect of sampling. This variation within sites was not greater than between sites and so did not influence site discrimination. When modelling bacterial and fungal diversity using calibrated log-likelihood ratios, we found samples were correctly predicted using dust 67% and 56% of the time and using soil 56% and 22% of the time for bacteria and fungi communities respectively. Incorrect predictions were related to within site variability, highlighting limitations to assigning dust provenance using metabarcoding of soil.


Hurricane Katrina: medical response at the Houston Astrodome/Reliant Center Complex.
On September 1, 2005, with only 12 hours notice, various collaborators established a medical facility--the Katrina Clinic--at the Astrodome/Reliant Center Complex in Houston. By the time the facility closed roughly two weeks later, the Katrina Clinic medical staff had seen over 11,000 of the estimated 27,000 Hurricane Katrina evacuees who sought shelter in the Complex. Herein, we describe the scope of this medical response, citing our major challenges, successes, and recommendations for conducting similar efforts in the future.


Exploration of synthetic lethal interactions as cancer drug targets.
In cancer research the quest continues to identify the Achilles' heel of cancer. The ideal cancer drug targets are those that are essential in tumor cells but not in normal cells. Such targets are defined as cancer-specific vulnerabilities or as synthetic lethal interactions with cancer-specific genetic lesions. The search for synthetic lethal interactions focuses on proteins that are frequently mutated but elude pharmacological inhibition, for example, RAS, or proteins that are lost in cancer cells and by definition cannot be targeted, such as the tumor suppressor genes p53, APC and RB. These genetic interactions could yield alternative, effective targets for cancer treatment. However, it remains very difficult to predict or extrapolate these synthetic lethal interactions based on existing knowledge. With the discovery of RNAi, unbiased large-scale functional genomic screens for the identification of such targets have become possible potentially leading to major advances in the treatment of cancers. In this review we will discuss the biological basis of synthetic lethal interactions in relation to existing targeted therapeutics, lessons taught by targeted therapeutics already used in the clinic and the implementation of RNAi as tool to identify such synthetic lethal interactions.


Clinical support role for a pharmacy technician within a primary care resource center.
The creation of a clinical support role for a pharmacy technician within a primary care resource center is described. In the Primary Care Resource Center (PCRC) Project, hospital-based care transition coordination hubs staffed by nurses and pharmacist teams were created in 6 independent community hospitals. At the largest site, patient volume for targeted diseases challenged the ability of the PCRC pharmacist to provide expected elements of care to targeted patients. Creation of a new pharmacy technician clinical support role was implemented as a cost-effective option to increase the pharmacist's efficiency. The pharmacist's work processes were reviewed and technical functions identified that could be assigned to a specially trained pharmacy technician under the direction of the PCRC pharmacist. Daily tasks performed by the pharmacy technician included maintenance of the patient roster and pending discharges, retrieval and documentation of pertinent laboratory and diagnostic test information from the patient's medical record, assembly of patient medication education materials, and identification of discrepancies between disparate systems' medication records. In the 6 months after establishing the PCRC pharmacy technician role, the pharmacist's completion of comprehensive medication reviews (CMRs) for target patients increased by 40.5% (p = 0.0223), driven largely by a 42.4% (p < 0.0001) decrease in the time to complete each chart review. The addition of a pharmacy technician to augment pharmacist care in a PCRC team extended the reach of the pharmacist and allowed more time for the pharmacist to engage patients. Technician support enabled the pharmacist to complete more CMRs and reduced the time required for chart reviews.


GOGO: An improved algorithm to measure the semantic similarity between gene ontology terms.
Measuring the semantic similarity between Gene Ontology (GO) terms is an essential step in functional bioinformatics research. We implemented a software named GOGO for calculating the semantic similarity between GO terms. GOGO has the advantages of both information-content-based and hybrid methods, such as Resnik's and Wang's methods. Moreover, GOGO is relatively fast and does not need to calculate information content (IC) from a large gene annotation corpus but still has the advantage of using IC. This is achieved by considering the number of children nodes in the GO directed acyclic graphs when calculating the semantic contribution of an ancestor node giving to its descendent nodes. GOGO can calculate functional similarities between genes and then cluster genes based on their functional similarities. Evaluations performed on multiple pathways retrieved from the saccharomyces genome database (SGD) show that GOGO can accurately and robustly cluster genes based on functional similarities. We release GOGO as a web server and also as a stand-alone tool, which allows convenient execution of the tool for a small number of GO terms or integration of the tool into bioinformatics pipelines for large-scale calculations. GOGO can be freely accessed or downloaded from http://dna.cs.miami.edu/GOGO/ .


Comprehensive and in-depth analysis of microRNA and mRNA expression profile in salivary adenoid cystic carcinoma.
To conduct an integrated analysis of microRNA and mRNA expression profile and further discover vital molecules to uncover novel pathogenic mechanisms in salivary adenoid cystic carcinoma (SACC). MicroRNA and mRNA expression profiles were obtained from six paired primary SACC tumors and corresponding adjacent normal glands using high-throughput next-generation sequencing technology followed by an overall integrated bioinformatics analysis and subsequently molecular biology techniques validation. Compared with adjacent noncancerous normal gland, 2107 significant differentially expressed mRNA were determined in SACC. Gene ontology and KEGG pathway analysis suggested that the differentially expressed genes were relevant to many significant biological implications. Venn diagram analysis of differentially expressed genes in different group identified 29 differentially expressed overlapping mRNA. 40 differentially expressed microRNAs were also identified in SACC. Furthermore, integrated analysis of microRNA and mRNA expression profiles recognized a core microRNA-mRNA regulatory network and unmasked many novel genes including SCUBE3, CA6, hsa-miR-885-5p and other molecules which may play an essential role in the carcinogenesis of SACC. Also, Q-PCR and immunohistochemistry results reveal the high expression and distribution of SCUBE3 in SACC and dual luciferase reporter assay also preliminarily validated that SCUBE3 was a target of hsa-miR-885-5p. Contemporary microRNA/mRNA analysis have uncovered many mRNAs and microRNAs worthy further exploration in SACC. These are bound to help us shed light on the overall genetic background of SACC and further elucidate the potential molecular mechanism of SACC.


Rational design based on bioactive conformation analysis of pyrimidinylbenzoates as acetohydroxyacid synthase inhibitors by integrating molecular docking, CoMFA, CoMSIA, and DFT calculations.
Pyrimidinylthiobenzoates constitute an important kind of herbicides targeting acetohydroxyacid synthase (AHAS, EC 126.96.36.199), which catalyze the first common step in branched-chain amino acid biosynthesis. Due to the symmetry of 4,6-dimethoxypyrimidyl, there are two kinds of conformation of pyrimidinylthiobenzoates: one's phenyl is left-extending (named conformation-L); the other's phenyl is right-extending (named conformation-R). On the basis of the assumption that 3D quantitative structure-activity relationship (QSAR) models derived from the bioactive conformation should give the best result, a strategy of density-functional-theory-based 3D-QSAR was proposed to identify the bioactive conformation of pyrimidinylthiobenzoates by integrating the techniques of molecular docking, comparative molecular field analysis (CoMFA), comparative molecular similarity indices analysis (CoMSIA), and density functional theory calculation. The combination of three criteria of q2, r2, and r2pred obtained from CoMFA and CoMSIA analyses clearly indicated that conformation-R rather than conformation-L might be the bioactive conformation for pyrimidinylthiobenzoates. A further comparison between the two binding modes indicated that pyrimidinylthiobenzoates and sulfonylureas have very similar binding sites, such as Trp586, Arg380, and Pro192. However, Lys251 formed H bonds with sulfonylureas rather than pyrimidinylthiobenzoates. In addition, the orientation of phenyl groups of the two classes of compounds in the binding pocket were revealed to be opposite, which explained why the mutation of Pro192 displayed different sensitivity to sulfonylureas and pyrimidinylthiobenzoates. On the basis of the understanding of interactions between pyrimidinyl-thiobenzoates and AHAS, we designed and synthesized six 8-(4,6-dimethoxypyrimidin-2-yloxy)-4-methylphthalazin-1-one derivatives according to the 3D-QSAR models. The excellent correlation between the tested Ki values against wild-type A. thaliana acetohydroxyacid synthase and the predicted IC50 values demonstrated the high reliability of the established 3D-QSAR models. To our knowledge, this is the first report highlighting the binding mode of herbicidal pyrimidinylthiobenzoates, which consisted of the reported results of herbicide resistance.


Associations between age, body size and nephron number with individual glomerular volumes in urban West African males.
Glomerulomegaly has been associated with an increased risk of renal disease. Few reports have investigated the heterogeneity of glomerular size within kidneys and associated risk factors. This study measured the individual glomerular volume (IGV) of 720 non-sclerotic glomeruli in kidneys of adult West African males, and investigated associations of IGV with age, total glomerular (nephron) number and body surface area (BSA). IGVs were determined in the kidneys of 24 Senegalese males from two age groups (12 subjects aged 20- 30 years and 12 subjects aged 50-70 years). Subjects were randomly chosen at autopsies performed at Le Dantec Hospital in Dakar. Volumes of 30 glomeruli per subject were determined using the disector/Cavalieri stereological method. IGVs ranged from 1.31 x 10(6) microm3 to 12.40 x 10(6) microm3 (a 9.4-fold variation). IGV varied up to 5.3-fold within single kidneys. The trimmed range of IGV within subjects (10th to 90th percentile of IGV) was directly correlated with median glomerular size. The mean and standard deviation (SD) of IGV did not differ significantly between age groups or between subjects with higher (> or =1.78 m2) and lower BSA (<1.78 m2). In older subjects the SD of IGV was significantly and directly correlated with BSA. Kidneys with less than 1 million nephrons had significantly larger mean IGV than kidneys with more than 1 million nephrons, and the trimmed range of IGVs within subjects was inversely correlated with total glomerular number. There was a considerable variation in IGV within kidneys of Senegalese males at autopsy. The heterogeneity of IGV was increased in association with low nephron number and increased BSA, with more pronounced effects in older subjects.


Quantifying the roles of conscious and unconscious processing in insight-related memory effectiveness within standard and creative advertising.
To maximize marketing effectiveness, many conscious and unconscious elements are simultaneously employed within campaign advertising. However, little is known about the individual contributions that conscious and unconscious processes make to the cognitive effectiveness of creative advertisements, some of which may also induce insight experiences. To quantify the roles of conscious and unconscious processes in memory effectiveness within commercial advertising, a dual-process, signal-detection technique was adopted to separate the contributions of conscious recollection and unconscious discrimination induced by 80 printed advertisements, among which half were considered standard and the other half creative. A total of 51 participants completed immediate (5 min later) and delayed (3 days later) memory recognition tests. In contrast to standard advertising, creative advertising was found to enhance recognition and to demonstrate advantages in both conscious and unconscious memory, which decreased across the test-time intervals. Further analyses showed that a moment of insight induced by an advertisement, regardless of whether it is standard or creative, can consolidate unconscious memory, whereas advertisements that do not induce insight improve conscious memory. The implications of these findings are discussed.


The relationship between physical fitness and ambulatory activity in very elderly women with normal functioning and functional limitations.
The effect of daily ambulatory activity on physical fitness has not yet been identified by quantitatively measuring the time spent on the intensity levels of ambulatory activity in elderly women over 75 with different functional capacity levels. The subjects consisted of 147 elderly women over 75 years old (82.8±4.3 years old) who were all capable of performing basic daily activities by themselves. Physical fitness was measured for 7 items (handgrip strength, knee extensor strength, postural stability, stepping, one-legged standing time with eyes open, 10 m walking, and the Timed Up and Go Test). The subjects wore a triaxial accelerometer for 2 consecutive weeks to measure their daily physical activities. The functional capacity level was assessed by the Tokyo Metropolitan Institute of Gerontology Index of Competence. The subjects were divided into two groups, a group with a score ≥10 points (high functional capacity group, n=59) and a score <10 points (low functional capacity group, n=88), and the relationship between physical fitness and physical activity was examined in both groups. In both the high and low functional capacity groups, 10 m walking, the Timed Up and Go Test, and one-legged standing time with eyes open significantly correlated with either the total steps/day or the ambulatory activity intensity. In the high functional capacity group, the knee extensor strength also significantly correlated with the total steps/day and moderate ambulatory activity. It is suggested that very elderly women with a reduced functional capacity should maintain their mobility by simply increasing their daily ambulatory activity.


Preferential recurrence of a sarcomatous component of a gliosarcoma after boron neutron capture therapy: case report.
Gliosarcoma, a rare pathological entity composed of 2-8% malignant gliomas, is characterized by a biphasic tissue pattern with alternating areas displaying glial and mesenchymal differentiation. Here we report the preferential recurrence of a sarcomatous component in gliosarcoma after boron neutron capture therapy (BNCT), while a gliomatous component disappeared as a result of the treatment. A 56-year-old woman with a left frontal tumor was introduced to our clinic. After stereotactic biopsy, craniotomy was applied and 90% of the mass was resected. The histological diagnosis was glioblastoma with small amounts of sarcomatous component, that is, gliosarcoma. BNCT was applied 30 days after craniotomy. Two weeks after BNCT, almost all of the contrast-enhanced mass had disappeared on magnetic resonance images; however, a half year later, the mass recurred just below the original site and extended posteriorly. Irrespective of repetitive salvage surgeries, the patient died of the recurrent tumor. At autopsy, tumor cells of the frontal lobe were absent. A well-circumscribed mass of the parietal and occipital lobes was composed of sarcomatous material, with very little glial fibrillary acid protein-positive glial material. We found in this patient the preferential recurrence of the sarcomatous component of a gliosarcoma after potent radiotherapeutics in the form of BNCT.


The effect of weight bearing on bone mineral density and bone growth in children with cerebral palsy: A randomized controlled preliminary trial.
The present study aims to explore the effect of weight bearing exercise on bone mineral density (BMD) and bone growth in children with cerebral palsy (CP). Twelve children with CP of functional level of gross motor functional classification scale (GMFCS) V and 6 healthy children (control group) were included in the study. Participants underwent a dual-energy X-ray absorptiometry scan to measure the BMD of the femur and full-length anteroposterior radiography to measure the bone length of the femur and tibia at baseline and after 6 months. Patients were randomly divided into 2 groups: group A with programmed standing exercises and assisted standing for more than 2 hours a day, more than 5 days a week; and group B with conventional physiotherapy with a standing program for 20 minutes a day, 2 to 3 days a week. A 6-month follow-up showed significantly increased BMD on the femur neck in the control group. Although the changes in BMD were not significant in both groups, group A demonstrated an increased trend of BMD, whereas group B showed a decreased trend. Bone length was significantly increased in all 3 groups at the 6-month follow-up. Although this increase was not significant, the change in bone length was greatest in the control group. The smallest changes were observed in group B. Weight bearing exercise may play an important role in increasing or maintaining BMD in children with CP and is also expected to promote bone growth. Programmed standing may be used as an effective treatment method to increase BMD in children with CP. However, further studies with a larger cohort and longer follow-up period are required to reveal further information on the benefit of weight bearing exercise and to develop a detailed program.


Effect of component distribution and nanoporosity in CuPt nanotubes on electrocatalysis of the oxygen reduction reaction.
Pt-based bimetallic electrocatalysts hold great potential in the oxygen reduction reaction (ORR) in current fuel-cell prototypes. However, they also face challenges from drastic dealloying of less-noble metals and coalescence of small nanoparticles. Porous and structure-ordered nanotubes may hold the potential to improve the stability of bimetallic electrocatalysts. Herein, we report a method to prepare CuPt nanotubes and porous Cu3 Pt intermetallic nanorods through a controlled galvanic replacement reaction and heat treatment process. The effect of the geometric features and compositional segregation on the electrocatalysis of the ORR was clarified. The outstanding performance of the Cu3 Pt/C-700 catalyst in the ORR relative to that of CuPt/C-RT was mainly attributed to the nanoporosity of the catalyst, whereas the enhanced specific activity on CuPt/C-RT after potential cycling was attributed to the interaction between the CuPt alloyed core and the Pt shell in the tube wall.


Monoclonal rheumatoid factor-secreting cells in a patient with mixed cryoglobulinemia. Homogeneity and stability of the idiotypic production and in vitro idiotypic suppression.
The potential therapeutic value of anti-idiotypic antibodies during B cell proliferations largely depends on the stability of the target Ig idiotopes. We investigated this stability in a clinical condition of so called nonmalignant monoclonal B cell proliferation, mixed cryoglobulinemia. The idiotypic profile of a single IgM kappa monoclonal auto-antibody with anti-IgG activity (rheumatoid factor (RF] which originated from a patient suffering from a nonmalignant mixed cryoglobulinemia was followed over a period of 3 yr. As judged from the reactivity of a panel of five different mouse monoclonal anti-idiotypic antibodies mapping the RF variable regions, there was no idiotypic change in the serum IgM RF. At a cellular level, in vitro stimulation of the patient's PBL gives rise to IgM kappa auto-antibodies that were shown to bear the same idiotypic determinants as the serum IgM kappa. We then investigated the effects of the anti-idiotypic antibodies on the in vitro IgM kappa production. When stimulated with PWM and in the presence of anti-idiotypic antibodies (10 micrograms/ml), the patient's PBL produced less IgM RF (18 to 62% inhibition). The same inhibition of IgM RF production was observed after EBV infection of the patient's PBL (from 19 to 90% inhibition). In both cases, the remaining IgM RF production was idiotypically indistinguishable from the serum IgM RF. The implications of the idiotypic stability and of the results of in vitro idiotypic manipulation could be important in view of both the understanding of nonmalignant cryoglobulinemia and of the possible therapeutic use of anti-idiotypic antibodies in diseases.


Prolonged immobilization and controlled ventilation do not improve outcome after global brain ischemia in monkeys.
This study is a therapeutic evaluation of prolonged immobilization and controlled intermittent positive-pressure ventilation (IPPV) after global brain ischemia (GBI) in pigtailed monkeys. Sixteen min of GBI was produced with a high-pressure neck cuff, while the lungs were being continuously ventilated. Normotension was restored within 2 min postischemia (PI). The control group of 13 monkeys was weaned from IPPV 4 to 6 h PI. The treatment group of 18 animals was paralyzed and ventilated with a 50:50 nitrous oxide-oxygen mixture for 48 h PI. Intensive care was maintained for 96 h PI. In the control group, 8 of 10 animals were awake at 96 h PI compared to 7 of 11 in the treatment group. Neurologic deficit scores for the survivors in the 2 groups were also not significantly different. Histologic examination supported this conclusion. Paralysis/IPPV for 48 h post-GBI with 50% N2O facilitates control of blood gases and blood pressure, but does not improve the neurologic outcome over that achieved with only 4 to 6 h controlled ventilation.


Targeting posttranslational modifications of RIOK1 inhibits the progression of colorectal and gastric cancers.
RIOK1 has recently been shown to play important roles in cancers, but its posttranslational regulation is largely unknown. Here we report that RIOK1 is methylated at K411 by SETD7 methyltransferase and that lysine-specific demethylase 1 (LSD1) reverses its methylation. The mutated RIOK1 (K411R) that cannot be methylated exhibits a longer half-life than does the methylated RIOK1. FBXO6 specifically interacts with K411-methylated RIOK1 through its FBA domain to induce RIOK1 ubiquitination. Casein kinase 2 (CK2) phosphorylates RIOK1 at T410, which stabilizes RIOK1 by antagonizing K411 methylation and impeding the recruitment of FBXO6 to RIOK1. Functional experiments demonstrate the RIOK1 methylation reduces the tumor growth and metastasis in mice model. Importantly, the protein levels of CK2 and LSD1 show an inverse correlation with FBXO6 and SETD7 expression in human colorectal cancer tissues. Together, this study highlights the importance of a RIOK1 methylation-phosphorylation switch in determining colorectal and gastric cancer development.


A system utilizing radio frequency identification (RFID) technology to monitor individual rodent behavior in complex social settings.
Pre-clinical investigation of human CNS disorders relies heavily on mouse models. However these show low predictive validity for translational success to humans, partly due to the extensive use of rapid, high-throughput behavioral assays. Improved assays to monitor rodent behavior over longer time scales in a variety of contexts while still maintaining the efficiency of data collection associated with high-throughput assays are needed. We developed an apparatus that uses radio frequency identification device (RFID) technology to facilitate long-term automated monitoring of the behavior of mice in socially or structurally complex cage environments. Mice that were individually marked and implanted with transponders were placed in pairs in the apparatus, and their locations continuously tracked for 24 h. Video observation was used to validate the RFID readings. The apparatus and its associated software accurately tracked the locations of all mice, yielding information about each mouse's location over time, its diel activity patterns, and the amount of time it was in the same location as the other mouse in the pair. The information that can be efficiently collected in this apparatus has a variety of applications for pre-clinical research on human CNS disorders, for example major depressive disorder and autism spectrum disorder, in that it can be used to quantify validated endophenotypes or biomarkers of these disorders using rodent models. While the specific configuration of the apparatus described here was designed to answer particular experimental questions, it can be modified in various ways to accommodate different experimental designs.


[Application of statistical simulation spectrophotometry to the determination of four medicaments in feibaosanxiao tablets].
The steps of statistical simulation spectrophotometry are to build a model based on a collection of limited but sufficient data from simulation samples to reveal the real absorbance-concentration relationship of the real sample and provide concentrated data of the individual components of the real sample measured by its absorbance data at several sensitive wavelengths. In this article, the method of stepwise regression has been used for building absorbance-concentration model of Feibaosanxiao tablets, a compound formulation of multimedicaments, and uniform design, for experimental design and recovery test. Recoveries of the four medicaments, acetaminophen, phenytoin sodium, theophylline and caffeine were 99.6%, 97.4%, 100.7% and 100.7% with RSD 0.3%, 1.9%, 1.3% and 1.3% respectively (n = 14).


[Clinical observation on treatment of diabetic nephropathy with Chinese drugs combined with benazepril].
To observe the protective effect on renal function of Chinese drugs combined with benazepril in patients with diabetic nephropathy (DN). Adopting stratified randomized and controlled design, 108 patients with DN were assigned to two groups. Besides the same measures of diet regulation, exercise, education, hypoglycemic and symptomatic treatment applied, benazepril was administered to all patients, and Chinese drugs were given to patients in the treated group additionally. The treatment course was 3 months. The changes before and after treatment of symptoms and signs, 24-h urinary protein (24 hUpro), serum creatinine (SCr), serum creatinine clearance rate (CCr), blood urea nitrogen (BUN), plasma glucose (PG) and blood pressure (BP) were observed, and the time of dialysis therapy initiating was recorded. SCr and 24 hUpro decreased and CCr increased in the treated group significantly (P < 0.01), and the improvement were superior to that in the control group (P < 0.05 or P < 0.01), the renal living time in the treated group was longer than that in the control group, but no significant different in BUN, PG and BP was found between the two groups (P > 0.05). The treatment program of Chinese drugs combined with benazepril could obviously reduce the levels of SCr and 24 hUpro, increase CCr in patients with DN, and its effect in improving renal function is better than that of using benazepril alone.


Clinical Outcomes and Complications during the Learning Curve for Reverse Total Shoulder Arthroplasty: An Analysis of the First 40 Cases.
The purpose of this study was to investigate the results and complications during the learning curve of reverse total shoulder arthroplasty (RTSA) for rotator cuff deficiency. We retrospectively reviewed the first 40 cases of RTSA performed by a single surgeon. The mean age of patients was 72.7 years (range, 63 to 81 years) and mean follow-up period was 26.7 months (range, 9 to 57 months). Clinical outcomes were evaluated using a visual analog scale (VAS) for pain, the University of California at Los Angeles (UCLA) shoulder score, American Shoulder and Elbow Surgeon (ASES) score, subjective shoulder value (SSV), and active range of motion (ROM). Intraoperative and postoperative complications were also evaluated. The average VAS pain score, UCLA score, ASES score, and SSV improved from 6.9%, 12.8%, 29.0%, and 29.0% before surgery to 1.6%, 27.0%, 73.3%, and 71.5% after surgery, respectively (p < 0.001). The mean forward flexion, abduction, and external rotation improved from 68.0°, 56.9°, and 28.0° before surgery to 131.0°, 112.3°, and 38.8° after surgery, respectively (p < 0.001, p < 0.001, and p = 0.021). However, the mean internal rotation did not improve after surgery (p = 0.889). Scapular notching was observed in 33 patients (51.5%). Eight shoulders (20%) had complications, including 2 major (1 deep infection and 1 glenoid fixation failure) and 6 minor complications (3 brachial plexus injuries, 2 acromial fractures, and 1 intraoperative periprosthetic fracture). The first 40 cases of RTSA performed by a single surgeon during the learning curve period showed satisfactory short-term follow-up results with an acceptable complication rate.


Weight loss in children and adolescents.
15% of children and adolescents in Germany are overweight, including 6.3% who are affected by obesity. The efficacy of conservative weight-loss treatments has been demonstrated, but there has not yet been a detailed analysis of their efficacy in terms of the amount of weight loss that can be expected. We re-evaluated the available evidence on this question, with particular attention to the methodological quality of clinical trials, in order to derive information that might be a useful guide for treatment. We conducted a systematic literature search of Medline for the period May 2008 (final inclusion date for a 2009 Cochrane Review) to December 2013. The identified studies were analyzed qualitatively. 48 randomized controlled clinical trials with a total of 5025 participants met the predefined inclusion criteria for this analysis. In the ones that met predefined criteria for methodological quality, conservative weight-loss treatments led to weight loss in amounts ranging from 0.05 to 0.42 BMI z score (standard deviation score of the body mass index) over a period of 12-24 months. Information on trial dropout rates was available for 41 of the 48 trials; the dropout rate was 10% or higher in 27 of these (66% ), and 25% or higher in 9 (22% ). The available evidence consistently shows that only a modest degree of weight loss can be expected from conservative treatment. Families seeking treatment should be informed of this fact. Future research should focus on determining predictive factors for therapeutic benefit, and on the evaluation of additional types of psychological intervention to promote coping with obesity.


Sphingobacterium prati sp. nov., isolated from agricultural soil and involved in lignocellulose deconstruction.
A bacterial strain, arapr2T, was isolated from agricultural soil sampled in Reims, France. Based on its 16S rRNA gene sequence, the strain was affiliated to the family Sphingobacteriaceae and more specifically to the genus Sphingobacterium. The strain had 98.31 % 16S rRNA gene sequence similarity to its closest relative Sphingobacterium canadense CR11T and 98.25 % to Sphingobacterium pakistanensis NCCP-246T. Genome relatedness indexes revealed that the average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between arapr2T and its closest relative (S. canadense CR11T) were 92.97 % and 52.00 %, respectively; for S. pakistanensis NCCP-246T, the ANI and dDDH values were 82.46 and 27.6%, respectively. The genomic DNA of strain arapr2T was 6.02 Mbp long, had a DNA G+C content of 40.4 mol% and had 5504 protein-coding genes. The results obtained in this study suggests that strain arapr2T (CIP 111872T=LMG 31848T) represents a new species for which the name Sphingobacterium prati sp. nov. is proposed. Due to the fact that this strain has been isolated using wheat straw as carbon source, this novel bacterial strain represents a promising biotechnological tool for the fractionation of lignocellulosic biomass in the context of biorefinery development.


Chromosomal Mapping of Repetitive DNA Sequences in the Genus Bryconamericus (Characidae) and DNA Barcoding to Differentiate Populations.
The mapping of repetitive DNA sites by fluorescence in situ hybridization has been widely used for karyotype studies in different species of fish, especially when dealing with related species or even genera presenting high chromosome variability. This study analyzed three populations of Bryconamericus, with diploid number preserved, but with different karyotype formulae. Bryconamericus ecai, from the Forquetinha river/RS, presented three new cytotypes, increasing the number of karyotype forms to seven in this population. Other two populations of Bryconamericus sp. from the Vermelho stream/PR and Cambuta river/PR exhibited interpopulation variation. The chromosome mapping of rDNA sites revealed unique markings among the three populations, showing inter- and intrapopulation variability located in the terminal region. The molecular analysis using DNA barcoding complementing the cytogenetic analysis also showed differentiation among the three populations. The U2 small nuclear DNA repetitive sequence exhibited conserved features, being located in the interstitial region of a single chromosome pair. This is the first report on its occurrence in the genus Bryconamericus. Data obtained revealed a karyotype variability already assigned to the genus, along with polymorphism of ribosomal sites, demonstrating that this group of fish can be undergoing a divergent evolutionary process, constituting a substantive model for studies of chromosomal evolution.


Endovascular approach to Leriche syndrome.
The chronic obstruction of the aortic bifurcation and iliac axis was definide by the French surgeon Rene Leriche, whose name it bears today. The advancing age of the population move the definition from a syndrome, as reported from Leriche, to complex aortoiliac vascular lesions current multidisciplinary guidelines recommend to treat extensive aortoiliac occlusive disease by surgical revascularization. Surgery provides good long-term patency, but at the cost of substantial perioperative morbidity. Development of new technologies and techniques has led to increased use of endovascular therapy for extensive aortoiliac disease. This review article summarized current multidisciplinary guidelines indication in endovascular treatment of extensive aorto-iliac disease and the various worldwide single centre experience published on endovascular treatment of Leriche Syndrome.


Closed-loop recyclable and biodegradable thioester-based covalent adaptable networks.
Closed-loop recyclable and biodegradable aliphatic covalent adaptable networks (CANs) based on dynamic β-CO thioester linkages that exhibit a service temperature beyond 100 °C are reported. These CANs possessing tensile strength and modulus values of up to 0.3 and 3 MPa, respectively, effectively undergo stress relaxation above 100 °C. The samples exhibit creep resistance ability and low hysteresis loss, and are repeatedly reprocessable at 120 °C. These CANs are depolymerizable to monomers under mild conditions and lose notable mechanical strength (92.4%) and weight (76.5%) within ∼35 days under natural biodegradation conditions.


Efficacy of duloxetine as a migraine preventive medication: possible predictors of response in a retrospective chart review.
This case series is a retrospective chart analysis conducted to evaluate the efficacy of duloxetine as a migraine preventive medication and to suggest possible predictors of response. Duloxetine, a relatively new selective serotonin and norepinephrine reuptake inhibitor, is FDA-approved for treatment of depression and diabetic peripheral neuropathic pain. The efficacy of duloxetine as a headache preventive medication is currently unknown. A retrospective chart review was performed using the electronic medical records of Headache Wellness Center, a headache specialty practice in Greensboro, North Carolina. From January 2004 to December 2006, 65 patients were identified who were prescribed duloxetine for migraine prevention for at least 2 months. Doses ranged from 30 mg qd to 90 mg qd. Frequency, severity, and impact of migraine disability were measured at baseline and compared to values obtained after 2 months of treatment. The total patient sample demonstrated a reduction in mean monthly headache frequency from 19.40 (SD=7.1) to 15.70 (SD=8.2) (P= .01). The 50% responder rate was 22%. In subset analysis, individuals with abnormal baseline Zung anxiety scores demonstrated a greater reduction in mean monthly headache frequency (4.28, P= .03) and a greater responder rate (25%) than those in the total patient sample. Non-statistically significant trends were observed in those patients with abnormal baseline Zung Depression scores exhibiting a less robust mean monthly migraine reduction (2.75, P= .18) than those with normal baseline depression scores (3.42, P= .07). Duloxetine demonstrates minimal effectiveness as a headache preventive medication. An interesting trend suggests that the presence of anxiety may be a positive predictor in treatment with duloxetine.


Long-term clinical and surrogate marker effects of subcutaneous intermittent interleukin-2 without antiretroviral therapy in HIV-infected patients.
Subcutaneous administration of interleukin-2 (IL-2) has been shown to increase CD4 counts in HIV-infected patients. It remains unclear whether this effect is associated with a clinical benefit. We conducted a long-term follow-up in the cohort of the UK-Vanguard study in which three groups of 12 antiretroviral-naive subjects with CD4 cell counts >350 cells/mm(3) received no treatment or IL-2 at either 4.5 or 7.5 MIU twice daily in 5 day cycles, respectively. Mean follow-up was 376 weeks. IL-2 therapy was associated with a higher area under the curve of CD4 cell count change from baseline at week 48 but not thereafter. HIV-RNA levels were unaffected. Highly active antiretroviral therapy (HAART) was initiated after a mean of 172, 175 and 152 weeks in the control group, low-dose and high-dose IL-2 treatment group, respectively, a statistically non-significant difference. There was a tendency to start HAART soon after discontinuation of IL-2 therapy which may have been triggered by the steep decay of CD4 counts. There were two serious adverse events in the control group, seven in the low-dose IL-2 group and eight in the high-dose IL-2 group. No pattern of disease was detected, making an association with IL-2 therapy unlikely. We could detect neither a benefit of IL-2 therapy after week 48 nor delayed initiation of HAART. This is currently the longest follow-up data comparing IL-2 therapy with no therapy in antiretroviral-naive HIV-infected patients and does not show a persistent benefit of the intervention.


[The impact of carbapenem-resistance Pseudomonas aeruginosa infections on mortality of patients with hematological disorders].
Objective: To assess the risk factors for mortality and clinical outcome of carbapenem-resistant Pseudomonas aeruginosa (CRPA) infections in patients with hematological disorders. Methods: The data of in-patients with hematological disorders infected by CRPA or carbapenem-susceptible Pseudomonas aeruginosa (CSPA) were recorded in a seven-year retrospective cohort study. Risk factors for CRPA infections and impact of on mortality were identified. The primary end point was 30-day all-cause mortality. Results: A total of 81 patients with PA infections were included in the study, including 58 CSPA and 23 CRPA. Most of the primary diseases were acute leukemia or lymphoma (79.0%, 64/81). The median absolute neutrophil count at infection onset was 0.24×10(9)/L. Independent risk factors associated with carbapenem-resistance included longer duration of hospital stay (P=0.013, OR=1.045) and carbapenem exposure one month prior to infections (P=0.005, OR=8.132). The 30-day all-cause mortality of the whole cohort was 29.6%(24/81), and 30-day attributable mortality was 13.6%(11/81). Pulmonary infection was the leading cause of death, accounting for 41.7%(10/24). The adjusted 30-day mortality rate was significantly higher in patients with CRPA compared with CSPA [60.9%(14/23) vs. 17.2%(10/58), P<0.001, respectively]. CRPA infection was an independent prognostic factor for 30-day mortality(P=0.011, OR=5.427). Other factors included old age, longer duration of neutropenia and poor functional performance. Conclusions: Patients with hematological disorders have high mortality rate and poor prognosis caused by CRPA infections, which mainly develop in lungs.


How dietary phosphate, renal failure and calcitriol administration affect the serum calcium-phosphate relationship in the rat.
The effect of hyperphosphataemia on serum calcium regulation in renal failure has not been well studied in a setting in which hypercalcaemia is not parathyroid hormone (PTH) mediated. In azotemic rats with a normal serum calcium concentration, an increased dietary phosphate burden affects serum calcium regulation because of its effects on skeletal resistance to PTH, calcitriol production, and possibly intestinal calcium absorption. Our goal was to determine how hyperphosphataemia affected the development of hypercalcaemia during calcitriol-induced hypercalcaemia and PTH suppression in azotemic rats with established hyperparathyroidism. Rats underwent a two-stage 5/6 nephrectomy or corresponding sham operations. After surgery, rats were given a high phosphate diet (P 1.2%) for 4 weeks to exacerbate hyperparathyroidism and were then changed to a normal diet (P 0.6%) for 2 weeks to normalize serum calcium values in the azotemic rats. At week 7, rats were divided into five groups and sacrificed after receiving three intraperitoneal doses of calcitriol (CTR, 500 pmol/100 g) or vehicle at 24 h intervals. The five groups and dietary phosphate content were: group 1, normal renal function (NRF)+0.6% P+vehicle; group 2, NRF+0.6% P+CTR; group 3, renal failure (RF)+0.6% P+vehicle; group 4, RF+1.2% P+CTR; and group 5, RF+0.6% P+CTR. Both the 0.6% and 1.2% phosphate diets contained 0.6% calcium. Serum creatinine values were increased (P<0.05) in 5/6 nephrectomized rats (groups 3, 4 and 5), as were serum calcium values (P<0.05) in CTR-treated rats (groups 2, 4 and 5) and serum phosphate values (P<0.05) in CTR-treated azotemic rats (groups 4 and 5). Serum PTH values were suppressed (P<0.05) in CTR-treated hypercalcemic rats (groups 2, 4 and 5) and increased (P<0.05) in azotemic rats not given CTR (group 3). In the azotemic groups (groups 3, 4 and 5), an inverse correlation was present between serum calcium and phosphate in each group, despite a wide variation in serum calcium values. The slope of the inverse relationship between serum calcium and phosphate was steeper in CTR-treated azotemic rats on a 1.2% phosphate (group 4) diet than on a 0.6% phosphate (group 5) diet (P=0.02). Thus, for a similar increase in the serum phosphate concentration, serum calcium values decreased more in group 4 than in group 5. The independent effect of dietary phosphate on serum calcium values was also confirmed by analysis of covariance. Finally, the serum calcium concentration was shown to be greater for any given serum phosphate value in CTR-treated rats than in those not on CTR. In azotemic rats with calcitriol-induced hypercalcaemia, the magnitude of hypercalcaemia is affected by: (i) the serum phosphate concentration; and (ii) differences in dietary phosphate content. Calcitriol administration also acts to shift upwards the relationship between serum calcium and phosphate so that a higher serum calcium concentration can be maintained for any given serum phosphate value.


Treatment of metastatic colorectal cancer with BRAF V600E mutation: A multicenter real-world study in China.
BRAF V600E mutant-metastatic colorectal cancer (mCRC) is characterized by its short survival time. Treatment approaches vary depending on whether or not the metastases are initially resectable. The benefit of metastasectomy remains unclear, and the optimal first-line treatment is controversial. This study aimed to describe the prognosis of BRAF V600E mutant-mCRC, analyze the recurrence pattern in resectable patients, and explore the optimal first-line treatment for unresectable patients. Patients diagnosed with BRAF V600E mutant-mCRC between February 2014 and January 2022 in five hospitals were enrolled. Date on clinical and pathological characteristics, treatment features, and survival outcomes were collected. Of the 220 included patients, 64 initially resectable patients had a significantly longer overall survival (OS) (37.07 vs. 20.20 months, P < 0.001) than initially unresectable patients. Of 156 unresectable patients, 54 received doublet (FOLFOX, XELOX or FOLFIRI) or triplet (FOLFOXIRI) chemotherapies (Chemo), 55 received Chemo plus Bevacizumab (Chemo+Bev), and 33 received vemurafenib plus cetuximab and irinotecan (VIC). The VIC regimen had a better progression-free survival (PFS) (12.70 months) than the Chemo (6.70 months, P < 0.001) and Chemo+Bev (8.8 months, P = 0.044) regimens. Patients treated with VIC had the best overall response rate (60.16%, P < 0.001), disease control rate (93.94%, P < 0.001) and conversional resection rate (24.24%, P = 0.003). Metastasectomy is beneficial to the survival of patients with BRAF V600E mutant-mCRC. For initially unresectable patients, VIC as first-line therapy is associated with a better prognosis and efficacy than doublet and triplet chemotherapy with or without bevacizumab.


Biosynthetic Pathway and Metabolic Engineering of Succinic Acid.
Succinic acid, a dicarboxylic acid produced as an intermediate of the tricarboxylic acid (TCA) cycle, is one of the most important platform chemicals for the production of various high value-added derivatives. As traditional chemical synthesis processes suffer from nonrenewable resources and environment pollution, succinic acid biosynthesis has drawn increasing attention as a viable, more environmentally friendly alternative. To date, several metabolic engineering approaches have been utilized for constructing and optimizing succinic acid cell factories. In this review, different succinic acid biosynthesis pathways are summarized, with a focus on the key enzymes and metabolic engineering approaches, which mainly include redirecting carbon flux, balancing NADH/NAD+ ratios, and optimizing CO2 supplementation. Finally, future perspectives on the microbial production of succinic acid are discussed.


High fat mass, low muscle mass, and arterial stiffness in a population of free-living healthy subjects: The "al passo con la tua salute" project.
A progressive decline in lean body mass and increase in fat mass occur with aging, and result in progressive weakness and impaired mobility; these features are altogether landmarks of the ageing frailty syndrome. High-fat mass and low muscle mass are both associated with an increased risk of cardiovascular events and are supposed to be risk factors for arterial stiffness. Little data analyzing the relationship between body composition and cardio-ankle vascular index (CAVI) are currently available. The main objective of this study was to verify whether low muscle mass and/or high fat mass could be associated with arterial stiffness measured by CAVI. Data are from the cross-sectional assessment of the "Al passo con la tua salute", a clinical study aimed to promote physical function among free-living elderly subjects.After a screening interview and a clinical visit aimed to exclude ineligible persons, 52 volunteers were enrolled in the study. All underwent: clinical examination, physical performance assessment, an interview on lifestyle and dietary habits, and lastly, a blood sample collection after at least 8 hours of fasting. CAVI was statistically significantly higher in those participants in the highest tertile of distribution for fat mass compared to all other subjects (P = .03). In those participants in the lowest tertile of distribution of muscle mass, compared to all other, CAVI was also statistically significant higher (P = .01) independently of age, sex, body mass index, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and systolic blood pressure. Low muscle mass and high fat mass were landmarks in the frailty model of aging; therefore, it is not inconsistent that both clinical conditions might share with the "aging vessel" a common pathway, probably mediated through signaling network deregulation and/or through alteration of the balance between energy availability and energy demand.


An 11-amino acid sequence in the cytoplasmic domain of CD40 is sufficient for activation of c-Jun N-terminal kinase, activation of MAPKAP kinase-2, phosphorylation of I kappa B alpha, and protection of WEHI-231 cells from anti-IgM-induced growth arrest.
We have previously shown that CD40 causes strong activation of the c-Jun N-terminal kinase (JNK), the p38 mitogen-activated protein kinases (MAPK) and MAPKAP kinase-2, a downstream target of p38 MAPK. To identify signaling motifs in the CD40 cytoplasmic domain that are responsible for activation of these kinases, we have created a set of 11 chimeric receptors consisting of the extracellular and transmembrane domains of CD8 fused to portions of the murine CD40 cytoplasmic domain. These chimeric receptors were expressed in WEHI-231 B lymphoma cells. We found that amino acids 35-45 of the CD40 cytoplasmic domain constitute an independent signaling motif that is sufficient for activation of the JNK and p38 MAPK pathways, as well as for induction of I kappa B alpha phosphorylation and degradation. Amino acids 35-45 were also sufficient to protect WEHI-231 cells from anti-IgM-induced growth arrest. This is the same region of CD40 required for binding the TNF receptor-associated factor-2 (TRAF2), TRAF3, and TRAF5 adapter proteins. These data support the idea that one or more of these TRAF proteins couple CD40 to the kinase cascades that activate NF-kappa B, JNK, and p38 MAPK.


A contribution to the diagnosis of Capillaria hepatica infection by indirect immunofluorescence test.
A highly specific pattern of immunofluorescence was noted when sera from Capillaria hepatica-infected rats were tested against the homologous worms and eggs present either in paraffin or cryostat sections from mouse liver. The pattern was represented by a combined apple green fluorescence of the internal contents of worms and eggs, which persisted in serum-dilutions of 1:400 up to 1:1600. Unequivocal fluorescent pattern was observed from 15 days up to 3 months following inoculation of rats with embryonated C. hepatica eggs and such result was confirmed by the ELISA. After the 4th month of infection, the indirect immunofluorescence test turned negative, probably revealing the extinction of parasitism, however the ELISA was contradictory, disclosing high levels of antibodies in this period. The IIF was also negative when control normal rat sera and sera from rats administered by gavage with immature C. hepatica eggs (spurious infection), or for reactions made against Schistosoma mansoni eggs, although a weakly positive pattern occurred with Fasciola hepatica eggs. The indirect immunofluorescence test may be recommended for use with human sera to detect early C. hepatica infection in special clinical instances and in epidemiological surveys, since it is a simple, inexpensive, and reliable test, presenting excellent sensitivity and specificity. Although the diagnosis is positive only during early infection, this is the period when the symptoms are usually more severe and the need for differential diagnosis is greater.


Myasthenia Gravis Induced by Immune Checkpoint Inhibitors.
Immune checkpoint inhibitors deeply improved the prognosis of metastatic melanoma or other types of cancer, but their related adverse effects (AEs) can be very severe, especially when the neurological system is touched, as in myasthenia gravis (MG). It is a rare immune AE that can be life-threatening and can be revealed by several symptoms. We report a case of our experience and review the current literature of MG exacerbated or occurring during immunotherapy to describe characteristics of this AE, warn the oncologist about this toxicity, and summarize the treatments conducted. Thirty-four cases of MG were reported, mostly with anti-programmed cell death protein 1 checkpoint inhibitor, and with melanoma. Onset was quick after the first or second infusion. Treatment comprised corticosteroids, prostigmine, and more or less plasmapheresis or immunoglobulins. Prognosis is poor, as 13 patients died after MG. MG is a rare immune-related AE that must be rapidly evoked and treated in case of neurological symptoms emerging after immunotherapy.


The role of PET in monitoring therapy.
Positron emission tomography (PET) is being increasingly used for the evaluation of patients with known or suspected cancer at all phases of the management process from diagnosis, through staging to follow-up after treatment. The role of PET in therapeutic monitoring is expanding rapidly due to its ability to provide earlier and more robust identification of non-responders than provided by conventional non-invasive imaging approaches. PET can thereby potentially provide important benefits to the individual patient by allowing an earlier change to alternative treatments that may be more efficacious or by avoiding the unnecessary toxicity related to ineffective therapy. As therapies become ever more expensive, this could also produce cost savings because of earlier termination of ineffective treatment. Conversely, PET may demonstrate important biological effects despite a lack of apparent morphological response and therefore prevent premature withdrawal of effective therapies. Globally, the vast majority of therapeutic monitoring studies use the glucose analogue, fluorine-18 fluorodeoxyglucose (FDG) but new tracers such as fluorine-18 fluorothymidine (FLT) also offer promise for this application. In this review, the potential benefits and limitations of FDG PET are discussed along with suggestions regarding the most practical methodologies for response evaluation using this modality.


sox4 and sox11 function during Xenopus laevis eye development.
SoxC genes are involved in many developmental processes such as cardiac, lymphoid, and bone development. The SoxC gene family is represented by Sox4, Sox11, and Sox12. Loss of either Sox4 or Sox11 function is lethal during mouse embryogenesis. Here, we demonstrate that sox4 and sox11 are strongly expressed in the developing eye, heart as well as brain in Xenopus laevis. Morpholino oligonucleotide mediated knock-down approaches in anterior neural tissue revealed that interference with either Sox4 or Sox11 function affects eye development. A detailed analysis demonstrated strong effects on eye size and retinal lamination. Neural induction was unaffected upon Sox4 or Sox11 MO injection and early eye field differentiation and cell proliferation were only mildly affected. Depletion of both genes, however, led independently to a significant increase in cell apoptosis in the eye. In summary, Sox4 and Sox11 are required for Xenopus visual system development.


Ethnic identity, spirituality, and self-efficacy influences on treatment outcomes among Hispanic American methadone maintenance clients.
There is increasing concern over whether standard health care services, such as substance abuse treatment, adequately account for important cultural influences, yet studies focusing on the impact of cultural influences on substance abuse treatment continue to be limited. The current study prospectively examined the effects of two cultural factors (ethnic identity and spirituality) on substance abuse treatment outcomes among Hispanic American clients (N=114) enrolled in methadone maintenance treatment. In addition, this study examined whether a commonly studied treatment factor (i.e., self-efficacy) significantly influenced treatment outcomes. Higher levels of self-efficacy at intake were related to increased odds of reported heroin abstinence and a lower number of drugs used at 1-year follow-up. Greater levels of ethnic identity were related to a greater number of drugs used at follow-up. No significant effects were found for spirituality.