CRISPR-based knock-out of eIF4E2 in a cherry tomato background successfully recapitulates resistance to pepper veinal mottle virus.
The host susceptibility factors are important targets to develop genetic resistances in crops. Genome editing tools offer exciting prospects to develop resistances based on these susceptibility factors, directly in the cultivar of choice. Translation initiation factors 4E have long been known to be a susceptibility factor to the main genus of Potyviridae, potyviruses, but the inactivation of the eIF4E2 gene has only recently been shown to provide resistance to some isolates of pepper veinal mottle virus (PVMV) in big-fruit tomato plants. Here, using CRISPR-Cas9-NG, we show how eIF4E2 can be targeted and inactivated in cherry tomato plants. Three independent knockout alleles caused by indel in the first exon of eIF4E2, resulted in the complete absence of the eIF4E2 protein. All three lines displayed a narrow resistance spectrum to potyvirus, similar to the one described earlier for an eIF4E2 EMS mutant of M82, a big-fruit tomato cultivar; the plants were fully resistant to PVMV-Ca31, partially to PVMV-IC and were fully susceptible to two isolates of PVY assayed: N605 and LYE84. These results show how easily a resistance based on eIF4E2 can be transferred across tomato cultivar, but also confirm that gene redundancy can narrow the resistances based on eIF4E knockout.


Introduction to the special section on the contribution of psychotherapy and pharmacotherapy research to national mental health care.
This special section represents a coordinated publication of the Journal of Consulting and Clinical Psychology and the Archives of General Psychiatry. These 2 journals have typically focused on somewhat different areas of research and have been read by different audiences. In an effort to achieve cross-fertilization of the disciplines of psychiatry and clinical psychology, this coordinated publication provides a current view of research both in psychopharmacology and psychotherapy as applied to major depression, anxiety disorders, alcohol abuse, and drug abuse. To further the aims of informing the diverse readership in areas to which they are often not exposed in the separate journals, the Journal of Consulting and Clinical Psychology is publishing in this section, articles that address biomedical and psychopharmacological research, and the Archives of General Psychiatry is publishing a coordinated set of articles on psychotherapy and behavior therapy. Collectively, the series contained in these 2 journals is designed both to foster knowledge and to enhance communication among psychiatrists and psychologists.


Future directions in the treatment of neuropathic pain: a review on various therapeutic targets.
Neuropathic pain is caused by structural lesion leading to functional abnormalities in central and peripheral nervous system. Neuropathic pain in itself is not always a disease, as it arises due to consequences of other diseases like diabetes, spinal cord injury, degenerative neuronal diseases and cancer. Current strategies of neuropathic pain treatment have provided relief to the patients to some extent, but complete cure is still a distant dream. In the future, it is hoped that a combination of new and improved pharmaceutical developments combined with careful clinical trials and increased understanding of neuroplasticity will lead to improved and effective pain management strategies leading to improved quality of life. In this review we have discussed various therapeutic targets of neuropathic pain and their pathophysiological mechanisms. Current status of drugs used for treatment of neuropathic pain have also been discussed in the review.


[Comparison of hang-back medial rectus recession with conventional recession for the correction of esotropia in children].
Extraocular muscles can be weakened by means of a conventional recession, or by means of a hang-back recession. This study compared the results of these techniques, their dose-response curves as well as intra- and postoperative complications for the correction of childhood esotropia. Healthy children with esotropia who received a medial rectus recession were included in this retrospective study. The recession was either performed as a conventional recession by direct scleral fixation, or as a hang-back recession. Ocular alignment (at distance fixation, 5 m and near fixation, 30 cm) as well as motility were compared preoperatively, and at 1 day and 3 months postoperatively. Dose-response curves and complication rates were compared. Seventy-four children from 2 to 17 years were enrolled. Forty-four children received a conventional medial rectus recession, thirty received a hang-back recession. Preoperatively, average esotropia was 33.5 ± 16.03 prism diopters (PD) at distance in the conventional group and 40.93 ± 18.02 PD in the hang-back group. At near, it was 38.73 ± 16.22 PD and + 45.93 ± 17.6 PD, respectively. Three months postoperatively, average distance esotropia was 6.41 ± 12.32 PD in the conventional group and 6.19 ± 10 PD in the hang-back group, average near esotropia was + 7.43 ± 12.10 PD in the conventional group and + 9.00 ± 11.78 PD in the hang-back group. On average, one millimeter of medial rectus recession corrected 3.01 ± 1.34 PD of distance esotropia und 3.57 ± 1.3 PD of near esotropia in the conventional group, and 3.26 ± 1.07 PD and 3.53 ± 1.15 PD in the hang-back group, respectively. This difference was not statistically significant (Mann-Whitney-U-Test, p > 0.05). Complications such as scleral perforation, slipped or lost muscles and induced vertical strabismus were not observed. Hang-back recession is an effective and safe alternative to conventional recession for the correction of esotropia in children. Larger prospective studies comparing long-term results of both techniques would be desirable.


A cautionary tale of sense-antisense gene pairs: independent regulation despite inverse correlation of expression.
Long non-coding RNAs (lncRNAs) have been proven to play important roles in diverse cellular processes including the DNA damage response. Nearly 40% of annotated lncRNAs are transcribed in antisense direction to other genes and have often been implicated in their regulation via transcript- or transcription-dependent mechanisms. However, it remains unclear whether inverse correlation of gene expression would generally point toward a regulatory interaction between the genes. Here, we profiled lncRNA and mRNA expression in lung and liver cancer cells after exposure to DNA damage. Our analysis revealed two pairs of mRNA-lncRNA sense-antisense transcripts being inversely expressed upon DNA damage. The lncRNA NOP14-AS1 was strongly upregulated upon DNA damage, while the mRNA for NOP14 was downregulated, both in a p53-dependent manner. For another pair, the lncRNA LIPE-AS1 was downregulated, while its antisense mRNA CEACAM1 was upregulated. To test whether as expected the antisense genes would regulate each other resulting in this highly significant inverse correlation, we employed antisense oligonucleotides and RNAi to study transcript-dependent effects as well as dCas9-based transcriptional modulation by CRISPRi/CRISPRa for transcription-dependent effects. Surprisingly, despite the strong stimulus-dependent inverse correlation, our data indicate that neither transcript- nor transcription-dependent mechanisms explain the inverse regulation of NOP14-AS1:NOP14 or LIPE-AS1:CEACAM1 expression. Hence, sense-antisense pairs whose expression is strongly-positively or negatively-correlated can be nonetheless regulated independently. This highlights the requirement of individual experimental studies for each antisense pair and prohibits drawing conclusions on regulatory mechanisms from expression correlations.


Preliminary efficacy trial of Cymelarsan in mice artificially infected with Trypanosoma brucei brucei isolated from a dog in Zambia.
An efficacy trial of Cymelarsan on a Zambian strain of Trypanosoma brucei brucei was done. Twenty-five male mice were infected intraperitoneally with 10(6) of T. b. brucei isolated from a dog. Five groups of 5 mice were treated with 0 (control), 0.25, 0.5, 1.0 and 2.0 mg/kg Cymelarsan, respectively. The target was to achieve aparasitaemia for 30 days post-treatment, euthanising those that remained parasitaemic or relapsed before then. The 0.25 and 0.5 mg/kg groups remained parasitaemic although the parasitaemic levels were reduced. The 1.0 mg/kg group had a proportion of aparasitaemic mice. However, all mice in the 2.0 mg/kg group remained aparasitaemic until day 20 when 2 mice relapsed. These results suggested that more than 2.0 mg/kg was required to eliminate this strain.


Critical Success Factors and Traceability Technologies for Establishing a Safe Pharmaceutical Supply Chain.
Drug counterfeits have been an international issue for almost two decades, and the latest statistics show that fake medications will continue to penetrate legitimate pharmaceutical supply chains (PSCs). Therefore, identifying the issues faced by PSCs is essential to combat the counterfeit drug problem, which will require the implementation of technologies in various phases of the PSC to gain better visibility. In this regard, a literature review was conducted to fulfill the following objectives: (i) review the application of traceability technologies in various PSC phases to detect counterfeits; (ii) analyze the various barriers affecting the establishment of a safe PSC and the critical success factors used to overcome those barriers; and (iii) develop a conceptual framework and guidelines to demonstrate the influence of traceability technologies and success factors on overcoming the various barriers in different phases of the PSC. The major finding of this review was that traceability technologies and the critical success factors have a significant influence on overcoming the barriers to establishing a safe PSC.


Theoretical study of interaction of urate with li(+), na(+), k(+), be(2+), mg(2+), and ca(2+) metal cations.
The geometries and energetics of complexes of Li(+), Na(+), K(+), Be(2+), Mg(2+), and Ca(2+)metal cations with different possible uric acid anions (urate) were studied. The complexes were optimized at the B3LYP level and the 6-311++G(d,p) basis set. Complexes of urate with Mg(2+), and Ca(2+)metal cations were also optimized at the MP2/6-31+G(d) level. Single point energy calculations were performed at the MP2/6-311++G(d,p) level. The interactions of the metal cations at different nucleophilic sites of various possible urate were considered. It was revealed that metal cations would interact with urate in a bi-coordinate manner. In the gas phase, the most preferred position for the interaction of Li(+), Na(+), and K(+) cations is between the N(3) and O(2) sites, while all divalent cations Be(2+), Mg(2+), and Ca(2+) prefer binding between the N(7) and O(6) sites of the corresponding urate. The influence of aqueous solvent on the relative stability of different complexes has been examined using the Tomasi's polarized continuum model. The basis set superposition error (BSSE) corrected interaction energy was also computed for complexes. The AIM theory has been applied to analyze the properties of the bond critical points (electron densities and their Laplacians) involved in the coordination between urate and the metal cations. It was revealed that aqueous solvation would have significant effect on the relative stability of complexes obtained by the interaction of urate with Mg(2+) and Ca(2+)cations. Consequently, several complexes were found to exist in the water solution. The effect of metal cations on different NH and CO stretching vibrational modes of uric acid has also been discussed.


Surface agents' influence on the flexural strength of bilaminated ceramics.
The objective of this study was to evaluate the influence of different surface agents on the flexural strength of a ceramic system. Eighty bar-shaped specimens of zirconia were divided into four groups according to the agent to be used: group Control--to be cleaned with alcohol; group VM9--application of a fluid layer of porcelain; group Effect Bonder--application of a bonding agent; and group Coloring Liquid--application of coloring liquid. All specimens received the porcelain application by the layering technique and were then subjected to thermocycling. The four-point bending test was performed to calculate the strength values (σ, MPa) and the failure modes were classified. ANOVA did not detect significant differences among the groups. The Weibull modulus were 5 (Control, VM9 and Effect Bonder) and 6 (Coloring Liquid). The cracking of the porcelain ceramic toward the interface was the predominant failure mode. It was concluded that the surface agents tested had no effect on the flexural strength of the bilaminated ceramic specimens.


Alien Invasion: Biology of Philornis Flies Highlighting Philornis downsi, an Introduced Parasite of Galápagos Birds.
The muscid genus Philornis comprises approximately 50 described species of flies, nearly all of which are obligate parasites of nestling birds. Philornis species are native to the Neotropics and widely distributed from Florida to Argentina. Most research on this group has focused on P. downsi, which was introduced to the Galápagos Islands in the late twentieth century. Although Philornis parasitism kills nestlings in several native host species, nowhere do the effects seem more severe than in P. downsi in the Galápagos. Here, we review studies of native and introduced Philornis in an attempt to identify factors that may influence virulence and consider implications for the conservation of hosts in the Galápagos.


Acute Exudative Polymorphous Paraneoplastic Vitelliform Maculopathy Managed With Intravitreal Aflibercept.
The authors report on two patients with bilateral acute exudative polymorphous paraneoplastic vitelliform maculopathy (AEPPVM) treated with intravitreal aflibercept (Eylea; Regeneron, Tarrytown, NY [marketed locally in Turkey by Bayer]). Underlying malignancy had been treated in each case, including breast carcinoma in one case and colon carcinoma in the other case. A macular vitelliform lesion was noted in the right eye and atrophic retinal pigment epithelial (RPE) changes were noted in the left eye of each case. Enhanced depth imaging optical coherence tomography (EDI-OCT) of the vitelliform lesion showed sensorineural retinal detachment, highly reflective subretinal material, ellipsoid loss in the right eye, and photoreceptor loss in both eyes of each patient. In both cases, the right eye with a vitelliform macular lesion was treated with intravitreal aflibercept (2.0 mg/0.05 mL) at monthly intervals for the first three injections and at bimonthly intervals for the following injections. Case 1 received a total of six injections and visual acuity (VA) increased from 20/70 to 20/50 at 10 months' follow-up. EDI-OCT showed slight gradual resolution of subretinal vitelliform material. Case 2 received three injections and VA increased from 20/100 to 20/40 at 4 months' follow-up with a decrease in the subretinal vitelliform deposit and intraretinal edema on EDI-OCT. Intravitreal aflibercept may control progression of APPVME in newly diagnosed cases by decreasing vascular leakage and stabilizing RPE function. [Ophthalmic Surg Lasers Imaging Retina. 2017;48:844-850.].


Induction of tumor necrosis factor-alpha mRNA in the brain after peripheral endotoxin treatment: comparison with interleukin-1 family and interleukin-6.
The constitutive expression of tumor necrosis factor-alpha (TNF alpha) mRNA and its induction (60 min later) by peripheral injection of Escherichia coli lipopolysaccharide (2 mg/kg i.p.) was demonstrated by polymerase chain reaction (PCR), in the pituitary and hypothalamus but not in the striatum or hippocampus of the rat. The pattern of TNF alpha mRNA induction is different from that observed for mRNAs of IL-1 alpha and IL-1 beta, IL-1ra and IL-6 respectively. This demonstration of the induction of TNF alpha in the brain may contribute to our understanding of the central effects of TNF alpha in fever and anorexia.


Examination of bilateral eyeblink conditioning in rats.
This experiment monitored eyelid responses bilaterally during delay eyeblink conditioning in rats. Rats were given paired or unpaired training with a tone or light conditioned stimulus (CS) and a unilateral periorbital shock unconditioned stimulus (US). Rats given paired training acquired high levels of conditioned responses (CRs), which occurred in both eyelids. However, acquisition was faster, and the overall percentage of CRs was greater in the eyelid that was ipsilateral to the US. CRs in the eyelid ipsilateral to the US also had shorter onset latencies and larger amplitudes than CRs in the contralateral eyelid. Both eyelids consistently showed high percentages of unconditioned responses (UR) to the US, and the UR amplitude decreased across training sessions in the paired group. The present study demonstrated that CRs occur robustly in both eyelids of rats given eyeblink conditioning, which is similar to previous findings in humans and monkeys. The results also showed that conditioning occurs more prominently in the eyelid that is ipsilateral to the US, which is similar to previous findings in humans, monkeys, dogs, and rabbits.


Control of mitotic exit in budding yeast. In vitro regulation of Tem1 GTPase by Bub2 and Bfa1.
The elimination of mitotic kinase activity at the end of mitosis is essential for progression to the next stage of the eukaryotic cell cycle. In budding yeast, this process is controlled by a regulatory cascade called the mitotic exit network. Extensive genetic data indicate that mitotic exit network activity is determined by a GTP-binding protein, Tem1, and its putative regulators, Bub2, Bfa1, and Lte1. Here we describe the purification and in vitro activities of Tem1, Bub2, and Bfa1. We describe the nucleotide binding properties of Tem1 and characterize its intrinsic GTPase activity. The combination of Bfa1 and Bub2 acts as a two-component GTPase-activating protein for Tem1. In the absence of Bub2, Bfa1 inhibits the GTPase and GTP exchange activities of Tem1. This inhibition is elicited by either the N- or C-terminal regions of Bfa1, which also retain some ability to co-activate GTPase activity in the presence of Bub2. Although the C-terminal region of Bfa1 binds to Bub2, no interaction of the N-terminal half of Bfa1 with Bub2 was detected despite their combined GAP activity. Therefore, we propose that Bfa1 acts both as an adaptor to connect Bub2 and Tem1 and as an allosteric effector that facilitates this interaction.


Neuropsychological deficits in older stroke patients.
This paper examines the frequency of CIND, the profile of early cognitive deficits in stroke patients, the differences in the profile of cognitive impairment in people with CIND and those with vascular dementia, and the MRI associations of CIND in older stroke patients.


Presentation of peptide antigens as albumin conjugates for use in detection of serum antibodies by enzyme-linked immunosorbent assay.
The use of linear peptides as antigens for detection of serum antibodies has been studied using a sequence of the Borrelia burgdorferi protein, flagellin, and Lyme disease sera as a model. It was found that a novel presentation of the peptide as a hapten on the carrier protein, bovine serum albumin, in the enzyme-linked immunosorbent assay format can be successfully applied to distinguish between Lyme disease and control sera.


[Diode laser cyclophotocoagulation--effect on intraocular pressure in comparison with cyclodiathermy under the scleral flap].
The aim of this study was to retrospectively analyze intraocular pressure and state of the eyeball in patients treated with diode laser, and to compare the outcomes with a group of patients after cyclodiahtermy under the scleral flap. We analyzed: visual acuity, IOP, percentage of lowering the IOP from the beginning, subjective patients' feelings of pain and complications. TSCPC is safe for patients and may be repeated, if needed. Laser cyclodestruction gives new possibilities in noninvasive IOP lowering with less complications than cyclodiahtermy, without changes in patient's visual functions.


Ecological modeling and pest population management: a possible and necessary connection in a changing world.
Ecological modeling is an important tool for investigating dynamic behavior patterns in populations, trophic interactions, and behavioral ecology. However, the ecological patterns that reflect population oscillation trends are often not clearly visible without analytical instruments such as ecological models. Thus, ecological modeling plays a fundamental role in describing demographic processes that are important for population dynamics. Ecological models, besides making possible the visualization of ecological patterns, may also reveal patterns of population persistence in many trophic systems, including prey-predator or host-parasitoid relationships, interactions that are commonly present in integrated pest management programs. In this forum, we present the main ecological aspects important for model building and implementation of integrated pest management programs for insects. Particularly, in this study, we analyze the combination between host-parasitoid models and the concept of economic threshold level on a spatio-temporal scale. As a conclusion about the model combination, spatial structure is essential for models of this nature, since its introduction into the system significantly alters the economic threshold-level values.


Retinoic acid is a potent regulator of growth plate chondrogenesis.
Vitamin A deficiency and excess both cause abnormalities in mammalian longitudinal bone growth. Because all-trans retinoic acid (RA) is synthesized from vitamin A, we hypothesized that RA regulates growth plate chondrogenesis. Consistent with this hypothesis, a single oral dose of RA reduced the height of the rat proximal tibial growth plate. To determine whether RA acts directly on growth plate, fetal rat metatarsal bones were cultured in the presence of RA. In this system, RA inhibited longitudinal bone growth by three mechanisms: 1) decreased chondrocyte proliferation, (assessed by 3H-thymidine incorporation), particularly in the proliferative zone of the growth plate; 2) decreased matrix synthesis (assessed by 35SO4 incorporation into glycosaminoglycans); and 3) decreased cell hypertrophy (determined histologically). The growth-inhibiting effects of RA were completely reversed by a retinoic acid receptor (RAR) antagonist. In the absence of exogenous RA, this antagonist accelerated bone growth, as did an RA-specific neutralizing antibody, suggesting that endogenous RA negatively regulates growth plate chondrogenesis. We conclude that RA, acting through RARs, negatively regulates longitudinal bone growth by inhibiting growth plate chondrocyte proliferation, chondrocyte hypertrophy, and matrix synthesis.


Common features of population climate in the Czech and Slovak Republics in 1991.
The author reports on a 1991 survey in the Czech and Slovak Republics "concerning the attitudes towards demographic tendencies, family formation and population-related policies.... The objective was to gain a better knowledge of the opinions and perceptions of the Czech and Slovak populations on these matters, as well as the demands to the government arising from society, both regarding its present position on the subject and its future responsibilities." Sections are included on family formation, the meaning of parenthood, and family policy and fertility intentions. (SUMMARY IN CZE)


Absolute determination of chemical kinetic rate constants by optical tracking the reaction on the second timescale using cavity-enhanced absorption spectroscopy.
We report a new spectroscopic platform coupled to an atmospheric simulation chamber for the direct determination of chemical rate constants with high accuracy at a second time-scale resolution. These developed analytical instruments consist of an incoherent broadband cavity enhanced absorption spectrometer using a red light emitting diode (LED) emitting at ∼662 nm (LED-IBBCEAS) associated with a multipass cell direct absorption spectrometer (MPC-DAS) coupled to an external cavity quantum cascade laser (EC-QCL) operating in the mid-infrared region at approximately 8 μm (EC-QCL-MPC-DAS). Spectrometers were employed to investigate the NO3-initiated oxidation of four selected volatile organic compounds (VOCs) for the determination of the corresponding rate constants with a dynamic range of 5 orders of magnitude (from 10-11 to 10-16 cm3 molecule-1 s-1). Rate constants of (6.5 ± 0.5) × 10-15, (7.0 ± 0.4) × 10-13, and (5.8 ± 0.5) × 10-16 cm3 molecule-1 s-1 for propanal, isoprene and formaldehyde, respectively, were directly determined by fitting the measured concentration-time profiles of NO3 and VOCs (measured using a proton transfer reaction time-of-flight mass spectrometer, PTR-ToF-MS) to chemical models based on the FACSIMILE simulation software (version 4.2.50) at 760 torr and 293 ± 2 K. The obtained rate constants are in good agreement with the most recent recommendations of the IUPAC (International Union of Pure and Applied Chemistry). In addition, a rate constant of (2.60 ± 0.30) × 10-11 cm3 molecule-1 s-1 for the oxidation of 2-methoxyphenol by NO3 radicals was first determined using the absolute kinetic method. Compared to the mostly used indirect relative rate method, the rate constant uncertainty is reduced from ∼20% to ∼12%. The results demonstrated the high potential of using modern spectroscopic techniques to directly determine the chemical reaction rate constants.


Cloning in Plasmid Vectors: Blunt-End Cloning.
This protocol describes procedures for cloning blunt-ended DNA fragments into linearized plasmid vectors. To obtain the maximum number of "correct" ligation products when cloning blunt-ended target fragments, the two components of DNA in the ligation reaction must be present at an appropriate ratio. If the molar ratio of plasmid vector to target DNA is too high, then the ligation reaction may generate an undesirable number of circular empty plasmids, both monomeric and polymeric; if too low, the ligation reaction may generate an excess of linear and circular homopolymers and heteropolymers of varying sizes, orientations, and compositions. For this reason, the orientation of the foreign DNA and the number of inserts in each recombinant clone must always be validated by restriction endonuclease mapping or some other means.


Multistage-Responsive Gene Editing to Sensitize Ion-Interference Enhanced Carbon Monoxide Gas Therapy.
As a promising therapeutic modality targeting cancer, gas therapy still faces critical challenges, especially in enhancing therapeutic efficacy and avoiding gas poisoning risks. Here, a pH/glutathione (GSH) dual stimuli-responsive CRISPR/Cas9 gene-editing nanoplatform combined with calcium-enhanced CO gas therapy for precise anticancer therapy, is established. In the tumor microenvironment (TME), the fast biodegradation of the CaCO3 layer via pH-induced hydrolyzation allows glucose oxidase (GOx) to catalyze glucose for H2 O2 production, which further reacts with manganese carbonyl (MnCO) and achieves the precise release of CO gas. Simultaneously, in situ Ca2+ overload from CaCO3 degradation disturbs mitochondrial Ca2+ homeostasis, resulting in Ca2+ -driven reactive oxygen species (ROS) formation and subsequent mitochondrial apoptosis signaling pathway activation. Subsequently, by GSH-induced cleavage of a disulfide bond, the released Cas9/sgRNA (RNP) can achieve nuclear factor E2-related factor 2 (Nrf2) gene ablation to sensitize gas therapy by interfering with ROS signaling. This therapeutic modality endows codelivery of CRISPR, ions, and gas with smart control features, which demonstrates great potential for future clinical applications in precise nanomedicine.


Genetic basis of thoracic aortic aneurysms and dissections.
The major diseases affecting the aorta are aortic aneurysms and dissections, which are classified based on anatomic location. Diseases affecting the ascending aorta, such as thoracic aortic aneurysms and type I and II dissections, are primarily associated with medial necrosis on pathologic examination. Medial necrosis is characterized by fragmentation and loss of elastic fibers, loss of smooth muscle cells, and interstitial collections of collagenous tissue and basophilic ground substance. Medial necrosis occurs as part of the normal aging of the aorta but is accelerated by other conditions, including hypertension and genetic alterations that predispose persons to these aortic diseases. The etiologies of many of the genetic syndromes, such as Marfan syndrome, that predispose persons to thoracic aortic aneurysms and dissections are understood. Studies are just beginning to elucidate the genes that predispose persons without known syndromes to these aortic diseases, and a major locus for this condition, termed the TAAD1 locus, has been mapped to 5q13-14. Future characterization of this gene and others will enhance the ability to determine persons at risk for aortic aneurysms and dissections and will define molecular mechanisms involved in the pathogenesis of this disorder.


The timing of preoperative prophylactic low-molecular-weight heparin administration in breast reconstruction.
Venous thromboembolism continues to be problematic despite increased recognition and advancements in venous thromboembolism prophylaxis. Although migration toward preoperative chemoprophylaxis increases, plastic surgeons seem reticent to adopt this practice. This study evaluates preoperative enoxaparin administration in breast reconstruction patients. Patients undergoing breast reconstruction performed by a single surgeon over a 5-year period were evaluated retrospectively. The authors introduced preoperative chemoprophylaxis with enoxaparin in all breast reconstructions during this time. Prosthetic-based and microsurgical breast reconstructions were examined. Patients were divided into two groups: those who did and those who did not receive preoperative enoxaparin. The authors reviewed patient demographics, comorbidities, and complications, focusing on bleeding complications. Three hundred patients (450 breasts) were included. One hundred fifty-four patients (244 breasts) underwent reconstruction with tissue expanders, and 146 (206 breasts) underwent free flap reconstructions. One hundred seventy-nine of 300 were given preoperative enoxaparin. Eleven hematomas occurred, eight (4.5 percent) in the enoxaparin group and three (2.5 percent) without enoxaparin (p = 0.399). Blood transfusions were given to four patients (2.2 percent) who received enoxaparin and one patient (0.8 percent) who did not (p = 0.652). Finally, any type of bleeding complication occurred in 11 patients (6.1 percent) with enoxaparin and in four (3.3 percent) without (p = 0.419). Larger breasts were more likely to receive enoxaparin (p = 0.011), which did not result in higher bleeding complications. In this retrospective study, the authors found that preoperative chemoprophylaxis in breast reconstruction was associated with an acceptable rate of postoperative bleeding complications. Therapeutic, III.


[Ca2+-activated K+ channels as cancer therapeutic targets].
Similar to calcium (Ca2+) and chloride (Cl-) ion channels/transporters, potassium (K+) channels have been recognized as a crucial cancer treatment target. Recent studies have provided convincing evidences of positive correlation between elevated expression levels of Ca2+-activated K+ (KCa) channels and cancer proliferation, metastasis, and poor patient prognosis. In cancer cells, KCa1.1 and KCa3.1 KCa channels are co-localized with Ca2+-permeable Orai/TRP channels to provide a positive-feedback loop for Ca2+ entry. They are responsible for the promotion of cell growth and metastasis in the different types of cancer, and are therefore potential therapeutic targets and biomarkers for cancer. We determined the epigenetic and post-transcriptional dysregulation of KCa3.1 by class I histone deacetylase inhibitors in breast and prostate cancer cells. We further determined the transcriptional repression and protein degradation of KCa1.1 by vitamin D receptor agonists and androgen receptor antagonists, which are expected as potential therapeutic drugs for triple-negative breast cancer. The anti-inflammatory cytokine, interleukin-10 (IL-10) is an immunosuppressive factor involved in tumorigenesis, and plays a crucial role in escape from tumor immune surveillance. We determined KCa3.1 activators are a possible therapeutic option to suppress the tumor-promoting activities of IL-10. These results may provide new insights into cancer treatment focused on Ca2+-activated K+ channels.


Intermediate accumulation of metabolites results in a bottleneck for mineralisation of the herbicide metabolite 2,6-dichlorobenzamide (BAM) by Aminobacter spp.
Degradation and mineralisation of the groundwater contaminant 2,6-dichloro-benzamide (BAM) was investigated in two Aminobacter strains focussing on the induction of BAM degradation and mineralisation and occurrence of intermediate metabolites. The BAM degradation rate was independent of whether the cells were pre-grown in the absence or presence of BAM, thus indicating that the first step in the degradation pathway was constitutively expressed. In contrast, (14)CO(2) production was stimulated when cells were pre-grown in the presence of BAM, suggesting that one or more of the subsequent steps in the degradation pathway were inducible. Accumulation of 2,6-dichlorobenzoate (DCBA) during degradation of BAM demonstrated that the first step involved amidase activity. Mass balance calculations and thin-layer chromatography coupled with autoradiographic detection indicated that degradation of DCBA and at least one unknown metabolite may comprise a bottleneck for BAM mineralisation by Aminobacter spp. The study thus provides novel information about the BAM degradation pathway and points to the involvement of unknown intermediate metabolites in degradation of this important groundwater contaminant.


Antibodies against H1N1 influenza virus hemagglutinin cross-react with prohibitin.
Influenza virus infection is associated with type 1 diabetes (T1DM), but its pathogenesis remains unclear. Here, our study found that one of the monoclonal antibodies against H1N1 influenza virus hemagglutinin(HA) cross-reacted with human pancreatic tissue and further demonstrated that it binded to rat islet β-cells. We immunoprecipitated islet protein with this cross-reactive antibody and identified the bound antigen as prohibitin by mass spectrometry. We then expressed the prohibitin protein in bacteria and confirmed the antibody binding to prohibitin by Western blot. We also verified the cross-reactivity of the antibody by prohibitin-siRNA transfection in islet beta cells. We conclude that prohibitin is an autoantigen that cross-reacts with influenza virus HA. The correlation between the autoantigen prohibitin and type 1 diabetes remains to be investigated.


The Rotterdam Study: 2018 update on objectives, design and main results.
The Rotterdam Study is a prospective cohort study ongoing since 1990 in the city of Rotterdam in The Netherlands. The study targets cardiovascular, endocrine, hepatic, neurological, ophthalmic, psychiatric, dermatological, otolaryngological, locomotor, and respiratory diseases. As of 2008, 14,926 subjects aged 45 years or over comprise the Rotterdam Study cohort. Since 2016, the cohort is being expanded by persons aged 40 years and over. The findings of the Rotterdam Study have been presented in over 1500 research articles and reports (see www.erasmus-epidemiology.nl/rotterdamstudy ). This article gives the rationale of the study and its design. It also presents a summary of the major findings and an update of the objectives and methods.


[Study of a typhoid fever outbreak in Baiona (Pontevedra)].
Despite of the fact of being a high endemic treat, the notification of a typhoid fever outbreak produces a very unusual epidemiological happening in our province. The uncommon of the sight and the importance of this process has made us focus on this study in order to understand its causes as well as to be able to establish some control according to them. In order to do that a retrospective epidemiological case-control study was made. Information was gathered about people who drank water from a public fountain in the village of Lages (Baiona) 14 cases and 52 controls randomly by hazard among the population of Lages and Almirante Fontán. The age mean of the people who became ill was between 25.6 +/- 16.9 years old and those control were 43.0 +/- 18.4 (p = 0.003). Among the 14 with the illness, 3 (21.4%) were men and 11 (78.6%) were women. The most frequent symptom was the fever (100%), followed by the cephaly (85.7%). The final OR was 57 (IC 95%: 13.2 - 246.2), being in Almirante Fontán a little higher (OR: 77; IC 95%: 4.7 - 572.1) than in Lages (OR: 40; IC 95%: 4.5 - 357.8). Salmonella Typhi strains was isolated in a 77.8% of the cases (7 of 9) where the blood culture was made. The water bacteriological analysis study proved a fecal contamination, but Salmonella Typhi strains wasn't isolated. The epidemiological and analytical suspicions guide us towards the water from the above mentioned fountain as the possible responsible for the coming about of this outbreak.


The comparative immunogenicity of human and porcine factor VIII in haemophilia A mice.
Inhibitory antibodies to factor VIII (FVIII inhibitors) are the most significant complication in the management of haemophilia A. The immunogenicity of FVIII may be driven in part by structural determinants within the FVIII molecule itself. Regions of nonidentity between human and porcine FVIII possibly could drive differential immune responses. The goal of this study was to compare the overall antibody response and levels of antibodies to the individual FVIII domains in naïve haemophilia A mice immunised with human or porcine FVIII. Haemophilia A mice were immunised with human or porcine FVIII using a protocol that mimics human clinical use. Inhibitor and total anti-FVIII antibody titers were measured and the domain-specificity of antibodies from 1,759 anti-FVIII hybridomas was determined. The overall immunogenicity of human and porcine FVIII was similar but significant differences in domain recognition were discovered. Anti-A2 and anti-C2 antibodies constituted the majority of inhibitors in both the human and porcine FVIII groups, similar to inhibitors that develop in humans. The proportions of anti-A2 or anti-C2 antibodies were not significantly different between the two groups. However, the specific inhibitory activity of anti-A2 antibodies was higher in the human FVIII group. Additionally, proportion of anti-C1 antibodies was significantly higher in the human FVIII group. In contrast, anti-A3 antibodies were more common in the porcine FVIII group. The differential immune response to human and porcine FVIII suggests that it may be possible to reduce the immunogenicity of FVIII by mutagenesis of the A2, A3 and C1 domains.


Dilated cardiomyopathy in children and adults: what is new?
Dilated cardiomyopathy (DCM) is the most common form of cardiomyopathy and cause of cardiac transplantation in children and young adults; mortality is high among this patient population. However, mortality, clinical course, and illustrative echocardiographic data of DCM in children and adults are not well established. Our objective was to provide a research article of detailed descriptions of the incidence, causes, outcomes, related risk factors, and new echocardiographic criteria of risk of death from DCM. Our results showed that independent risk factors at DCM diagnosis for subsequent death or transplantation in children cohorts were older age, congestive heart failure, lower left ventricular ejection fraction (EF< or =25%), low global strain, significant mitral valve incompetence, pulmonary hypertension, diastolic dysfunction, right ventricular involvement, and cause of DCM (p<0.001 for all). In adults, low ejection fraction (<30-35%), global peak systolic strain <-7.6%, increased EDV, ESV, LBBB, diastolic dysfunction, and left ventricle dyssynchrony were the main independent risk factors for major cardiac events and need for CRT or transplantation (p<0.001 for all). Our conclusions were that in children and adults, DCM is a diverse disorder with outcomes that depend largely on cause, age, heart failure status at presentation, and echocardiographic parameters of the heart (systolic and diastolic function of left ventricle, pulmonary artery pressure, global strain, and valvular function of the mitral valve). This study will present new findings in the diagnostic area.


L-O-(2-malonyl)tyrosine: a new phosphotyrosyl mimetic for the preparation of Src homology 2 domain inhibitory peptides.
Inhibition of Src homology 2 (SH2) domain-binding interactions affords one potential means of modulating protein-tyrosine kinase-dependent signaling. Small phosphotyrosyl (pTyr)-containing peptides are able to bind to SH2 domains and compete with larger pTyr peptides or native pTyr-containing protein ligands. Such pTyr-containing peptides are limited in their utility as SH2 domain inhibitors in vivo due to their hydrolytic lability to protein-tyrosine phosphatases (PTPs) and the poor cellular penetration of the ionized phosphate moiety. An important aspect of SH2 domain inhibitor design is the creation of pTyr mimetics which are stable to PTPs and have reasonable bioavailability. To date, most PTP-resistant pTyr mimetics which bind to SH2 domains are phosphonates such as (phosphonomethyl)phenylalanine (Pmp, 2), [(monofluorophosphono)methyl]phenylalanine (FPmp, 3) or [(difluorophosphono)methyl]-phenylalanine (F2Pmp, 4). Herein we report the incorporation of a new non-phosphorus-containing pTyr mimetic, L-O-(2-malonyl)tyrosine (L-OMT, 5), into SH2 domain inhibitory peptides using the protected analogue L-N alpha-Fmoc-O'-(O",O"-di-tert-butyl-2-malonyl)tyrosine (6) and solid-phase peptide synthesis techniques. Five OMT-containing peptides were prepared against the following SH2 domains: the PI-3 kinase C-terminal p85 SH2 domain (Ac-D-(L-OMT)-V-P-M-L-amide, 10, IC50 = 14.2 microM), the Src SH2 domain (Ac-Q-(L-OMT)-E-E-I-P-amide, 11, IC50 = 25 microM, and Ac-Q-(L-OMT)-(L-OMT)-E-I-P-amide, 14, IC50 = 23 microM), the Grb2 SH2 domain (Ac-N-(L-OMT)-V-N-I-E-amide, 12, IC50 = 120 microM), and the N-terminal SH-PTP2 SH2 domain (Ac-L-N-(L-OMT)-I-D-L-D-L-V-amide, 13, IC50 = 22.0 microM). These results show that peptides 10, 11, 13, and 14 have reasonable affinity for their respective SH2 domains, with the IC50 value for the SH-PTP2 SH2 domain-directed peptide 13 being equivalent to that previously observed for the corresponding F2Pmp-containing peptide. OMT may afford a new structural starting point for the development of novel and useful SH2 domain inhibitors.


Longitudinal study of bone age in acute lymphoblastic leukaemia.
Bone age was assessed in children with acute lymphoblastic leukaemia (ALL) using the Tanner-Whitehouse II method. X-rays of the left hand (wrist) were made at diagnosis and annually until 5 years after diagnosis. A total of 164 X-rays from 40 patients was available. During treatment bone age development as well as height growth was retarded in relation to calendar age. A catch-up of bone age and height was observed in the 2 years after cessation of treatment. No significant differences could be found between patients with and without cranial irradiation as central nervous system treatment. Therefore cranial irradiation alone is not responsible for the growth retardation of children with ALL; the causative role of cytostatic drug therapy and/or corticosteroids has to be investigated.


Polybrominated diphenyl ether (PBDE) accumulation by earthworms (Eisenia fetida) exposed to biosolids-, polyurethane foam microparticle-, and Penta-BDE-amended soils.
Polybrominated diphenyl ether (PBDE) flame retardants have been used in consumer polymers at up to percent levels. While long viewed as biologically inaccessible therein, PBDEs may become bioaccessible following volatilization or polymer deterioration. PBDEs may then enter soils via polymer fragmentation or following land application of sewage sludge-derived biosolids. Studies of direct PBDE uptake from these materials by soil organisms are scarce. We thus exposed earthworms ( Eisenia fetida ) to artificial soil amended with a Class B anaerobically digested biosolid (ADB), an exceptional quality composted biosolid (CB), PBDE-containing polyurethane foam (PUF) microparticles, and Penta-BDE-spiked artificial soil (SAS). Worms accumulated mg/kg (lipid) ∑Penta-PBDE burdens from all substrates. Biota-soil accumulation factors (BSAFs) for worms exposed to ADB- and CB-amended soils were comparable after 28 d. BSAFs generally decreased with increasing congener KOW and substrate dosage. Biosolids-associated PBDE bioavailability was lower than spiked PBDEs. BSAFs for worms exposed to PUF microparticles ranged from 3.9 to 33.4, with ∑Penta-PBDE tissue burdens reaching 3740 mg/kg lipid. Congener accumulation patterns were similar in worms and polyethylene passive sampling devices immersed in ADB-amended soil coincident with exposed worms. However, passive sampler accumulation factors were lower than BSAFs. Our results demonstrate that PBDEs may accumulate in organisms ingesting soils containing biosolids or waste plastics. Such organisms may then transfer their burdens to predators or translocate them from the site of application/disposal.


Glucose-6-phosphate dehydrogenase blockade potentiates tyrosine kinase inhibitor effect on breast cancer cells through autophagy perturbation.
Glucose-6-phospate dehydrogenase (G6PD) is the limiting enzyme of the pentose phosphate pathway (PPP) correlated to cancer progression and drug resistance. We previously showed that G6PD inhibition leads to Endoplasmic Reticulum (ER) stress often associated to autophagy deregulation. The latter can be induced by target-based agents such as Lapatinib, an anti-HER2 tyrosine kinase inhibitor (TKI) largely used in breast cancer treatment. Here we investigate whether G6PD inhibition causes autophagy alteration, which can potentiate Lapatinib effect on cancer cells. Immunofluorescence and flow cytometry for LC3B and lysosomes tracker were used to study autophagy in cells treated with lapatinib and/or G6PD inhibitors (polydatin). Immunoblots for LC3B and p62 were performed to confirm autophagy flux analyses together with puncta and colocalization studies. We generated a cell line overexpressing G6PD and performed synergism studies on cell growth inhibition induced by Lapatinib and Polydatin using the median effect by Chou-Talay. Synergism studies were additionally validated with apoptosis analysis by annexin V/PI staining in the presence or absence of autophagy blockers. We found that the inhibition of G6PD induced endoplasmic reticulum stress, which was responsible for the deregulation of autophagy flux. Indeed, G6PD blockade caused a consistent increase of autophagosomes formation independently from mTOR status. Cells engineered to overexpress G6PD became resilient to autophagy and resistant to lapatinib. On the other hand, G6PD inhibition synergistically increased lapatinib-induced cytotoxic effect on cancer cells, while autophagy blockade abolished this effect. Finally, in silico studies showed a significant correlation between G6PD expression and tumour relapse/resistance in patients. These results point out that autophagy and PPP are crucial players in TKI resistance, and highlight a peculiar vulnerability of breast cancer cells, where impairment of metabolic pathways and autophagy could be used to reinforce TKI efficacy in cancer treatment.


The Cognitive Underpinnings of Multiply-Constrained Problem Solving.
Individuals encounter problems daily wherein varying numbers of constraints require delimitation of memory to target goal-satisfying information. Multiply-constrained problems, such as the compound remote associates, are commonly used to study this type of problem solving. Since their development, multiply-constrained problems have been theoretically and empirically related to creative thinking, analytical problem solving, insight problem solving, and a multitude of other cognitive abilities. In the present study, we empirically evaluated the range of cognitive abilities previously associated with multiply-constrained problem solving to assess common versus unique predictive variance (i.e., working memory, attention control, episodic and semantic memory, and fluid and crystallized intelligence). Additionally, we sought to determine whether problem-solving ability and self-reported strategy adoption (analytical or insightful) were task specific or task general through the use of novel multiply-constrained problem-solving tasks (TriBond and Location Bond). Performance across these tasks was shown to be domain general, solutions derived through insightful strategies were more often correct than those derived through analytical strategies, and crystallized intelligence was the sole cognitive ability that provided unique predictive value after accounting for all other abilities.


External ear nodule revealing a disseminated Kaposi disease.
Kaposi disease (KD) is an angiogenetic tumor process, characterized by its various clinical aspects. Its pathogenesis is multifactorial and it was attributed to HHV-8 infection. We report an original case of disseminated KD revealed by solitary lesion of external ear in a patient treated by corticosteroids for bullous pemphigoid.


Interferon-Beta Therapy of Multiple Sclerosis Patients Improves the Responsiveness of T Cells for Immune Suppression by Regulatory T Cells.
Multiple sclerosis (MS) is an inflammatory autoimmune disease characterized by imbalanced immune regulatory networks, and MS patient-derived T effector cells are inefficiently suppressed through regulatory T cells (Treg), a phenomenon known as Treg resistance. In the current study we investigated T cell function in MS patients before and after interferon-beta therapy. We compared cytokine profile, responsiveness for Treg-mediated suppression ex vivo and evaluated reactivity of T cells in vivo using a humanized mouse model. We found that CD4+ and CD8+ T cells of therapy-naive MS patients were resistant to Treg-mediated suppression. Treg resistance is associated with an augmented IL-6 production, enhanced IL-6 receptor expression, and increased PKB/c-Akt phosphorylation. These parameters as well as responsiveness of T cells to Treg-mediated suppression were restored after interferon-beta therapy of MS patients. Following transfer into immunodeficient mice, MS T cells induced a lethal graft versus host disease (GvHD) and in contrast to T cells of healthy volunteers, this aggressive T cell response could not be controlled by Treg, but was abolished by anti-IL-6 receptor antibodies. However, magnitude and lethality of GvHD induced by MS T cells was significantly decreased after interferon-beta therapy and the reaction was prevented by Treg activation in vivo. Our data reveals that interferon-beta therapy improves the immunoregulation of autoaggressive T effector cells in MS patients by changing the IL-6 signal transduction pathway, thus restoring their sensitivity to Treg-mediated suppression.


Guinea pig erythrocytes, after their contact with influenza virus, acquire the ability to activate the human alternative complement pathway through virus-induced desialation of the cells.
Guinea pig erythrocytes that had been exposed to influenza A virus activated the alternative complement pathway in whole human serum in the absence of natural antibodies. Because all virus particles were eluted from the treated cells, activation was not dependent on antiviral antibodies or on virus particles themselves. The relative capacity of treated erythrocytes to activate the alternative pathway was dependent on the amount of virus to which the cells had been exposed and was directly related to the amount of sialic acid removed from the erythrocyte membrane during incubation with either whole virus particles or purified viral sialidase. C3b bound to cells that had been treated with virus, and P-stabilized amplification convertase sites P,C3b,Bb formed on these cells, exhibited increased resistance to the action of the regulatory proteins beta-1H and C3b Ina compared with C3b and P,C3b,Bb on untreated, nonactivating cells. The acquired resistance of the cell-bound, P-stabilized amplification convertase to decay-dissociation by beta-1H was directly related to the activating capacity of the treated cells in whole serum (r = 0.95) and to the amount of sialic acid removed from the cells by the virus (r = 0.98). Desialation represents a specific alteration of the cell surface by which a nonimmune host, through activation of the alternative pathway, may deposit C3b on a target cell that had been exposed to influenza virus and may lyse virus virus-modified cells during orthomyxovirus infections.


A negative MYB regulator of proanthocyanidin accumulation, identified through expression quantitative locus mapping in the grape berry.
Flavonoids are secondary metabolites with multiple functions. In grape (Vitis vinifera), the most abundant flavonoids are proanthocyanidins (PAs), major quality determinants for fruit and wine. However, knowledge about the regulation of PA composition is sparse. Thus, we aimed to identify novel genomic regions involved in this mechanism. Expression quantitative trait locus (eQTL) mapping was performed on the transcript abundance of five downstream PA synthesis genes (dihydroflavonol reductase (VvDFR), leucoanthocyanidin dioxygenase (VvLDOX), leucoanthocyanidin reductase (VvLAR1), VvLAR2 and anthocyanidin reductase (VvANR)) measured by real-time quantitative PCR on a pseudo F1 population in two growing seasons. Twenty-one eQTLs were identified; 17 of them did not overlap with known candidate transcription factors or cis-regulatory sequences. These novel loci and the presence of digenic epistasis support the previous hypothesis of a polygenic regulatory mechanism for PA biosynthesis. In a genomic region co-locating eQTLs for VvDFR, VvLDOX and VvLAR1, gene annotation and a transcriptomic survey suggested that VvMYBC2-L1, a gene coding for an R2R3-MYB protein, is involved in regulating PA synthesis. Phylogenetic analysis showed its high similarity to characterized negative MYB factors. Its spatiotemporal expression profile in grape coincided with PA synthesis. Its functional characterization via overexpression in grapevine hairy roots demonstrated its ability to reduce the amount of PA and to down-regulate expression of PA genes.


Exploring Strategies to Improve Adherence to Immunization Schedule: A Study among Children Attending Maternal and Child Health Clinic at Kenyatta National Hospital, Nairobi, Kenya.
Globally, immunization is among the major contributors to public health, preventing 20% of childhood mortality annually. The highest fatality rates from vaccine preventable diseases are usually among children under five. Despite immunization guidelines put in place by the World Health Organization, globally, 1.5 million children die annually related to inadequate vaccination coverage. Existing literature indicate that there is an increase in nonadherence to immunization schedule in developing countries, and therefore, there is an increased demand to improve adherence to immunization schedule. To explore strategies that will improve adherence to immunization schedule among children under 24 months attending the Maternal and Child Health clinic at Kenyatta National Hospital. A cross-sectional mixed method study involving caregivers (n = 214) of well babies attending the Maternal and Child Health clinic. Data was collected using semistructured questionnaires, focus group discussions, and key informant interviews and analyzed using SPSS V.20. There was a significant relationship between the level of education and marital status of the caregivers and adherence to immunization schedule. Barriers found that is related to adherence to immunization schedule included far distance from health facility, baby's sickness, and vaccine stock-outs while employment of a caregiver was a constrainer factor. The enabling factors to current strategies of improving adherence to immunization schedule were having more health facilities near residential areas, using text messages reminders a day before the clinic date to remind caregivers of the due date for the clinic, and constant availability of vaccines. The health system strategies that would improve adherence to immunization schedules were more flexible clinic hours, availability of vaccines on daily basis, phone call reminders by health care providers, and increasing awareness on the importance of both vaccinations and adherence to immunization schedule.


Inverse relation of autocrine motility factor receptor and E-cadherin expression following MDCK epithelial cell transformation.
Autocrine motility factor interacts with its cell surface receptor (AMF-R) to stimulate tumor cell motility. To study AMF-R expression following transformation of polarized epithelial MDCK cells, we have used the invasive Moloney sarcoma virus transformed MDCK (MSV-MDCK) cell population. Decreased E-cadherin expression of the transformed MSV-MDCK clones is associated with both increased cellular motility and increased AMF-R expression. Increased AMF-R expression is due to MSV transformation as differentially motile MSV-MDCK clones, which either retain low E-cadherin, exhibit equivalent high levels of AMF-R. Loss of the polarized epithelial phenotype and increased cellular motility following transformation of MDCK cells is thus associated with a shift from a high E-cadherin/low AMF-R to a low E-cadherin/low AMF-R phenotype.


Leptin: a pivotal mediator of intestinal inflammation in mice.
In addition to acting as a regulator of food intake and energy expenditure, leptin can also modulate immune and inflammatory responses. The role of leptin in intestinal inflammation is the focus of the present study. Acute and chronic colitis were induced in leptin-deficient ob/ob or wild-type (WT) mice using dextran sulfate sodium (DSS) or trinitrobenzene sulfonic acid (TNBS). The severity of colitis was evaluated, and possible mechanisms were studied. Leptin directly stimulates intraepithelial lymphocytes (IELs) and lamina propria mononuclear cells (LPMCs). In the DSS acute model, ob/ob mice exhibited a 72% reduction of colitis severity and spontaneous release of proinflammatory cytokines from the colon compared with WT mice. Replacement of leptin in ob/ob mice converted disease resistance to susceptibility, indicating that leptin deficiency, not obesity, accounts for the resistance to acute DSS-induced colitis. During chronic DSS-induced colitis and TNBS-induced colitis, in addition to reduced disease severity, ob/ob mice exhibited a significant attenuation in intestinal inflammation, accompanied by reduced production of cytokines and chemokines. When compared with WT mice, CD8(+) IELs of ob/ob mice were reduced in number as well as in their ability to synthesize interferon gamma. In addition, LPMCs of ob/ob mice showed increased apoptosis in untreated as well as DSS- or TNBS-treated mice. Phosphorylation of signal transducer and activator of transcription 3 and induction of cyclooxygenase 2 were absent in the colon of DSS-fed ob/ob mice. These results show that leptin represents a functional link between the endocrine and immune systems.


Prognostic significance of disseminated tumor cells in the connective tissue of patients with medullary thyroid carcinoma.
Disseminated tumor cells in the connective tissue (CT-DTCs) do not have any connection to a primary tumor or the lymph nodes. They are identified quite often in patients with medullary thyroid carcinoma (MTC), but nothing is known regarding their prognostic significance. Among 450 patients with MTC, 69 (15%) were identified as having CT-DTCs. A case-control group of patients without CT-DTCs was selected. The two groups were matched concerning TNM classification, age, heredity, and sex. Because many patients with CT-DTCs had extrathyroidal tumor extension (pT4 category), distant metastases (M1 category), or both, only 35 matched pairs could be identified. The TNM classification in both groups was as follows: pT1, n = 8; pT2, n = 15; pT3, n = 4; pT4, n = 8; pN0, n = 4; pN1, n = 31; M0, n = 30; M1, n = 5. The mean age was 46.8 +/- 17.0 years in the CT-DTC group and 44.4 +/- 15.0 years in the case-control group (NS). In both groups, 23 patients had sporadic MTC, and 12 patients had hereditary MTC. Neither mean basal preoperative nor postoperative calcitonin levels differed significantly between the two groups. In contrast, none of the patients with CT-DTCs was biochemically cured (normal calcitonin level after pentagastrin stimulation) compared to eight patients without CT-DTCs (P < 0.005). The two groups did not differ concerning other parameters (basal calcitonin level > 3000 pg/ml, more than 10 lymph node metastases, more than two involved locoregional lymph node compartments, mediastinal lymph node metastases) that have been reported to correlate with the lack of or almost (< 10%) lack of biochemical cure. In patients with MTC, disseminated tumor cells in the connective tissue correlate with advanced tumor stages and appear to be of prognostic significance.


Facilitative effect of a glutamate transporter inhibitor (2S,3S)-3-{3-[4-(trifluoromethyl)benzoylamino]benzyloxy}aspartate on the expression of methamphetamine-induced behavioral sensitization in rats.
We examined the effects of a potent glutamate transporter inhibitor, (2S,3S)-3-{3-[4-(trifluoromethyl)benzoylamino]benzyloxy}aspartate (TFB-TBOA), on the expression of methamphetamine-induced behavioral sensitization in rats. Rats were intraperitoneally treated with 2 mg/kg methamphetamine for 5 days and then challenged with 1 mg/kg methamphetamine. Intracerebroventricular administration of TFB-TBOA (0.1 nmol) 10 min before the challenge significantly facilitated the expression of behavioral sensitization. On the other hand, it had no effect on the locomotor activation elicited by the challenge with methamphetamine in repeated-saline-treated (non-sensitized) rats. These results suggest that central glutamate transporters may play an inhibitory role in the expression of behavioral sensitization to methamphetamine.


Antimicrobial and plaque inhibitory potential of herbal and probiotic oral rinses in children: a randomized clinical trial.
Antimicrobial agents serve as an effective adjunct with mechanical means in plaque control. Chlorhexidine has been the gold standard in the field of dentistry, but these days a growing number of dentists are embracing the philosophy that natural agents are better for children's oral health, and the focus is shifted toward biogenic agents for oral hygiene maintenance in children. The aim was to evaluate antimicrobial and plaque inhibitory potential of herbal and probiotic rinses against Streptococcus viridans with commonly used antimicrobial agent like 0.2% chlorhexidine digluconate. A randomized clinical trial was conducted on 60 subjects aged between 6 and 14 years and were divided into three groups comprising 20 subjects in each group. Three oral rinses were administered twice daily for a period of 1 week. Estimation of plaque scores and S. viridans counts was done before and after intervention, and the results were statistically analyzed. The change in mean plaque index in Groups A, B, and C was 0.28 ± 0.16, 1.37 ± 0.43, and 0.60 ± 0.35 respectively. Furthermore, change in mean log 10 colony forming unit (CFU)/ml of S. viridans in Groups A, B, and C was 0.13 ± 0.06, 0.53 ± 0.17, and 0.22 ± 0.06 CFU/ml, respectively. Based on observations done during the course of study herbal rinse proved equally effective as 0.2% chlorhexidine digluconate in reducing S. viridans counts and plaque accumulation after 1 week of intervention, whereas probiotic rinse was least effective. However, long-term clinical trial with larger sample size needs to be undertaken, especially to evaluate beneficial effects of biogenic agents such as herbal and probiotic rinses.


Human scFv antibody fragments specific for the epithelial tumour marker MUC-1, selected by phage display on living cells.
New anti-cancer agents are being developed that specifically recognise tumour cells. Recognition is dependent upon the enhanced expression of antigenic determinants on the surface of tumour cells. The tumour exposure and the extracellular accessibility of the mucin MUC-1 make this marker a suitable target for tumour diagnosis and therapy. We isolated and characterised six human scFv antibody fragments that bound to the MUC-1 core protein, by selecting a large naive human phage display library directly on a MUC-1-expressing breast carcinoma cell line. Their binding characteristics have been studied by ELISA, FACS and indirect immunofluorescence. The human scFv antibody fragments were specific for the tandem repeat region of MUC-1 and their binding is inhibited by soluble antigen. Four human scFv antibody fragments (M2, M3, M8, M12) recognised the hydrophilic PDTRP region of the MUC-1 core protein, which is thought to be an immunodominant region. The human scFv antibody fragments were stable in human serum at 37 degrees C and retained their binding specificity. For imaging or targeting to tumours over-expressing MUC-1, it might be feasible to use these human scFv, or multivalent derivatives, as vehicles to deliver anti-cancer agents.


Multiple births in Nigerian Igbo women: incidence and outcomes.
The rate of multiple births varies amongst races and ethnic groups. This study shows that the incidence of multiple births among the Nigerian Igbo women is one in 35.5 or 28.2 per 1000 births. A significant difference in Apgar scores at 1 minute between the first and second twins suggests a greater risk for the second twin following the delivery of the first twin. Significantly lower Apgar scores and higher perinatal mortality rate in the twins compared with the singleton controls also indicate greater hazard for the multiple births in general. Successful management of multiple pregnancy can occur following early diagnosis and confirmation of fetal presentations, adequate antepartum fetal surveillance and prompt management of pregnancy complications; appropriate referrals to specialised centres when indicated; and prompt decisions as to timing and mode of delivery.


7-Benzyloxyquinoline oxidation by P450eryF A245T: finding of a new fluorescent substrate probe.
The main objective of the present study was to find a fluorescent substrate probe for cytochrome P450eryF (P450eryF). P450eryF is a bacterial P450 that catalyzes the hydroxylation of 6-deoxyerythronolide B at the 6S position, a necessary step in the biosynthesis of erythromycin. The lack of a conserved threonine residue in the I-helix, in contrast to other P450s, makes P450eryF unable to oxidize other substrates. A recent study [Xiang et al. (2000) J. Biol. Chem. 275, 35999-36006] has shown that the substitution of Ala-245 by threonine confers on P450eryF significant testosterone hydroxylase activity. Therefore, we investigated various known fluorescent P450 substrates with P450eryF wild-type as well as two mutants, A245S and A245T. Among the various fluorescent compounds tested, 7-benzyloxyquinoline (7-BQ) was found to be the most suitable probe for P450eryF A245T, with rates of oxidation being lower for A245S and wild-type enzyme. The steady-state kinetics of 7-BQ oxidation by A245T are sigmoidal (V(max) = 0.71 nmol/min/nmol, n = 2.18, and S(50) = 132 microM). alpha-Naphthoflavone (alpha-NF), a well-known activator of CYP3A4, did not stimulate 7-BQ oxidation by A245T, although the S(50) value for alpha-NF binding to wild-type P450eryF was similar to P450 3A4. Interestingly, spectral binding studies of wild-type P450eryF and A245T with ketoconazole and miconazole showed differential binding behaviors. Titration of wild-type with ketoconazole and miconazole and of A245T with miconazole showed the expected type-II binding. However, titration of A245T with ketoconazole produced a spectrum similar to type-I. Inhibition studies showed that both ketoconazole and miconazole are able to inhibit 7-BQ oxidation by A245T, although miconazole showed a slightly higher potency. In brief, the present study reports the discovery of 7-BQ as the first fluorescent and only the second unnatural substrate, and of miconazole as an effective P450eryF inhibitor.


Fulminant course in a case of malignant phyllodes tumor.
We present the case of a 31-year-old woman with an inflammatory and ulcerative malignant phyllodes tumor in her right breast. A right modified radical mastectomy and transverse rectus abdominis myocutaneous (TRAM) flap were performed. A month after the initial operation, several masses recurred at the superior margin and deep margin of the TRAM flap. Wide excision was performed, but masses recurred at the inferior margin and in both lung fields 2 weeks after the second operation. Six weeks after the second operation, the patient died due to progression of dyspnea and respiratory failure.


Novel 1,4-benzothiazine derivatives as large conductance Ca2+-activated potassium channel openers.
The design and synthesis of a novel class of 1,4-benzothiazines targeted for the large-conductance calcium-activated potassium channels (BK) are presented. In vitro functional characterization of BK channel opening activity was assessed by measuring the relaxation of isolated rat aortic rings precontracted with KCl 20 mM. The results of this study show that the 1,4-benzothiazine heterocyclic nucleus is a suitable backbone for designing novel BK-openers; indeed, some of these new 1,4-benzothiazine derivatives had a vasorelaxant potency comparable or superior to that of reference BK-activator NS-1619 (1).


Development of type 2 diabetes in children and adolescents.
Type 2 diabetes is emerging as a significant health problem in children and adolescents. The disease usually develops in obese insulin-resistant youth with a typical pattern of lipid partitioning characterized by increased lipid deposition in myocytes, the visceral compartment, and the liver. Unfavorable adipocytokine profiles, together with a state of low-grade inflammation, create an additional metabolic burden tightly coupled to other components of the metabolic syndrome. Insufficient beta-cell compensation promotes altered glucose metabolism.


Axial and radial segregation of granular mixtures in a rotating spherical container.
We report the formation of axial segregation patterns of bidisperse granular mixtures of glass beads in a spherical container which is rotated about a central axis. When the rotation axis is horizontal, three distinct segregation bands are formed for a broad range of geometrical and dynamical parameters. We find two distinct regimes: at low fill levels the small size beads of the mixture are collected at the poles and the large size beads in a central band. At high fill levels the pole regions are occupied by large beads, while the small beads form the central band. The critical fill level for this structural transition decreases with increasing container size. For a container with 37 mm inner diameter, containing a mixture of 0.5 and 1.5 mm beads, the transition occurs between 40% and 50% fill level. When the rotation axis is tilted, the band positions are shifted and two-band structures are formed with the smaller particles at the lower pole. In our experiments the granulate is submersed in water; this allows NMR imaging of the complete three-dimensional band structures. We compare the observed segregation structures to those in cylindrical mixers and propose a model for the qualitative explanation of the pattern formation process.


Relaxed active space: fixing tailored-CC with high order coupled cluster. II.
Due to the steep increase in computational cost with the inclusion of higher-connected cluster operators in coupled-cluster applications, it is usually not practical to use such methods for larger systems or basis sets without an active space partitioning. This study generates an active space subject to unambiguous statistical criteria to define a space whose size permits treatment at the CCSDT level. The automated scheme makes it unnecessary for the user to judge whether a chosen active space is sufficient to correctly solve the problem. Two demanding applications are presented: twisted ethylene and the transition states for the bicyclo[1,1,0]butane isomerization. As bi-radicals both systems require at least a CCSDT level of theory for quantitative results, for the geometries and energies.


Computed tomography in the management of blunt abdominal trauma in children.
Sixteen children with suspected abdominal injury were reviewed. All underwent plain abdominal radiography and computed tomography (CT). Fifteen CT scans showed an abnormality, and the abdominal radiograph was abnormal in seven cases. Eight patients also underwent abdominal ultrasonography, and six scans failed to demonstrate an abnormality. Only two patients required laparotomy, one for a ruptured left diaphragm and one for a major renal injury. These results support the use of CT as the optimal method for assessing the stable child following blunt abdominal trauma.


Inhibition of Fas-mediated apoptosis through administration of soluble Fas receptor improves functional outcome and reduces posttraumatic axonal degeneration after acute spinal cord injury.
Fas receptor activation has been implicated in inflammatory responses, programmed cell death, Wallerian degeneration in neural injury and the axotomy induced death of motoneurons. Recent work using transection models of spinal cord injury (SCI) demonstrated that neutralization of Fas ligand with antibodies may promote axonal regeneration and functional recovery. Moreover, recent studies from our laboratory in mutant mice with deficient expression of Fas, show reduced cell death and enhanced behavioural recovery after SCI. The present paper examines the effects of soluble Fas receptor (sFasR) administration on inhibition of Fas receptor-Fas ligand interaction in the setting of acute SCI in vitro and in vivo. An in vitro model of SCI demonstrated that sFasR administration decreases cell death as assessed by propidium iodide fluorescence. Furthermore, in a moderately severe in vivo clip compression model of SCI at C7-T1, we demonstrate that subarachnoid infusion of sFasR results in increased neuron and oligodendrocyte survival, improved tissue and long tract axonal preservation, reduced apoptotic cell death and enhanced functional neurological outcome after acute SCI. These results strongly suggest that inhibiting Fas receptor activation is neuroprotective after acute SCI and that this strategy may have important translational significance.


Joint Women's Programme v. State of Rajasthan, 24 March 1987.
By means of this decision, the Supreme Court of India directed the states of Rajasthan and Haryana to proceed with the investigation of two so-called dowry deaths. The decision also directed the two states to create Special Dowry Cells at the state level to investigate dowry deaths through special investigative units and authorized the ministry dealing with social welfare and women's problems to designate one or two leading women social workers to be associated with the Cells.


Genotypic diversity and antibiotic resistance in Sphingomonadaceae isolated from hospital tap water.
The aim of this study was to infer about the modes and extent of dispersion of Sphingomonadaceae via tap water. Sphingomonadaceae isolated from tap water samples in different places of a hospital were compared, based on intra-species genetic variability and antibiotic resistance phenotypes. These isolates were also compared with others isolated before from houses and dental chairs, served by the same municipal water supply system. Sphingomonadaceae from hospital tap water comprised members of the genera Sphingomonas, Sphingobium, Novosphingobium and Blastomonas. In general, distinct genotypes of Sphingomonadaceae were detected in different hospital areas and in tap water outside the hospital, suggesting these bacteria are not persistent or widespread in the urban water distribution system. Possible intrinsic antibiotic resistance, observed in most or all members of the family or of a genus, was observed for colistin in Sphingomonadaceae, aminoglycosides in the genus Blastomonas and beta-lactams in the genus Sphingobium. Possible acquired resistance phenotypes, not common to all members of a given species, comprised fluoroquinolones, cephalosporins and sulphonamides. Although the potential of Sphingomonadaceae as opportunistic pathogens may be low, the capacity of these bacteria to thrive in water supply systems, combined with the intrinsic or acquired antibiotic resistance, may raise the risk associated with their occurrence in hospital tap water.


Childhood adherence to a potentially healthy and sustainable Nordic diet and later overweight: The Norwegian Mother, Father and Child Cohort Study (MoBa).
The New Nordic Diet (NND) is a potentially healthy and sustainable dietary pattern represented by locally available and traditionally consumed foods in the Northern countries. The diet has been commonly examined in adult populations, but less is known regarding its potential associations with overweight/obesity in children. We have previously developed child diet scores measuring compliance to the NND at child age 6 and 18 months and 3 and 7 years. In this study, we aimed to describe child and maternal characteristics and assess potential associations between the age-specific diet scores and child overweight at 8 years. This study is based on the Norwegian Mother, Father and Child Cohort Study (MoBa), including 14,989 mother-child pairs and uses data from the Medical Birth Registry of Norway (MBRN). The scores measured NND compliance as a total score and categorized into low, medium and high NND compliance at each age point. Using logistic regression models, we investigated the association between each age-specific score and the odds of overweight at 8 years. In crude analyses, adherence to the NND at 6 months was inversely associated with odds of overweight at 8 years in the continuous score (odds ratio = 0.95, 95% CI [0.91, 0.98]) and when comparing high versus low NND adherence (odds ratio = 0.81, 95% CI [0.70, 0.94]). The association was almost entirely attenuated in the adjusted models. In conclusion, child NND adherence up to 7 years of age was not associated with odds of overweight at 8 years in adjusted analyses.


Marsupial genetics and genomics.
Marsupials, the 'other' mammals, are found only in Australasia and the Americas. They are quite different from eutherian ('placental') mammals, as well they might be after 130 million years of separate evolution. They display a unique pattern of mammalian organization and development that is reflected by differences in their genomes. Here, we introduce marsupials as alternative (but not inferior!) mammals and summarize the state of knowledge of marsupial relationships, marsupial chromosomes, maps, genes and genetic regulatory systems. We shamelessly present the case for a Kangaroo Genome Project.


Financial Loss for Inpatient Care of Medicaid-Insured Children.
Medicaid payments tend to be less than the cost of care. Federal Disproportionate Share Hospital (DSH) payments help hospitals recover such uncompensated costs of Medicaid-insured and uninsured patients. The Patient Protection and Affordable Care Act reduces DSH payments in anticipation of fewer uninsured patients and therefore decreased uncompensated care. However, unlike adults, few hospitalized children are uninsured, while many have Medicaid coverage. Therefore, DSH payment reductions may expose extensive Medicaid financial losses for hospitals serving large absolute numbers of children. To identify types of hospitals with the highest Medicaid losses from pediatric inpatient care and to estimate the proportion of losses recovered through DSH payments. This retrospective cross-sectional analysis evaluated Medicaid-insured hospital discharges of patients 20 years and younger from 23 states in the 2009 Kids' Inpatient Database. The dates of the analysis were March to September 2015. Hospitals were categorized as freestanding children's hospitals (FSCHs), children's hospitals within general hospitals, non-children's hospital teaching hospitals, and non-children's hospital nonteaching hospitals. Financial records of FSCHs in the data set were used to estimate the proportion of Medicaid losses recovered through DSH payments. Hospital financial losses from inpatient care of Medicaid-insured children (defined as the reimbursement minus the cost of care) were compared across hospital types. For our subsample of FSCHs, Medicaid-insured inpatient financial losses were calculated with and without each hospital's DSH payment. The 2009 Kids' Inpatient Database study population included 1485 hospitals and 843 725 Medicaid-insured discharges. Freestanding children's hospitals had a higher median number of Medicaid-insured discharges (4082; interquartile range [IQR], 3524-5213) vs non-children's hospital teaching hospitals (674; IQR, 258-1414) and non-children's hospital nonteaching hospitals (161; IQR, 41-420). Freestanding children's hospitals had the largest median Medicaid losses from pediatric inpatient care (-$9 722 367; IQR, -$16 248 369 to -$2 137 902). Smaller losses were experienced by non-children's hospital teaching hospitals (-$204 100; IQR, -$1 014 100 to $14 700]) and non-children's hospital nonteaching hospitals (-$28 310; IQR, -$152 370 to $9040]). Disproportionate Share Hospital payments to FSCHs reduced their Medicaid losses by almost half. Estimated financial losses from pediatric inpatients covered by Medicaid were much larger for FSCHs than for other hospital types. For children's hospitals, small anticipated increases in insured children are unlikely to offset the reductions in DSH payments.


Adhesion of Bordetella pertussis to eukaryotic cells requires a time-dependent export and maturation of filamentous hemagglutinin.
Bordetella pertussis, the human pathogen of whooping cough, when grown at 22 degrees C is nonvirulent and unable to bind eukaryotic cells. In response to a temperature shift to 37 degrees C, the bacterium acquires the ability to bind eukaryotic cells in a time-dependent fashion. By studying in vitro the temperature-induced transition, from the nonvirulent to the virulent state, we found that binding to CHO cells is mediated by the Arg-Gly-Asp-containing domain of filamentous hemagglutinin (FHA), a protein with multiple binding specificities. This protein is synthesized as a 367-kDa polypeptide within 10 min after temperature shift, but requires 2 hr before it is detected on the bacterial cell surface and starts to bind CHO cells. Mutations affecting the cell surface export of FHA abolish bacterial adhesion to CHO cells, while mutations in the outer membrane protein pertactin strongly reduce binding. This suggests that multiple chaperon proteins are required for a correct function of FHA. Finally, several hours after maximum binding efficiency is achieved, the N-terminal 220-kDa portion of FHA that contains the binding regions is cleaved off, possibly to release the bacteria from the bound cells and facilitate spreading. The different forms of FHA may play different roles during bacterial infection.


Population pharmacokinetic analysis of tocilizumab in patients with rheumatoid arthritis.
Tocilizumab is a humanized anti-interleukin-6 (IL-6) receptor monoclonal antibody that has demonstrated efficacy in the treatment of rheumatoid arthritis (RA). A population pharmacokinetic (PK) model was developed using nonlinear mixed effect modeling to describe the PK profile of tocilizumab and used to estimate interindividual variability and assess the influence of covariates on PK parameters. The model was constructed based on data collected from 1793 patients with moderate to severe RA who received tocilizumab (4 or 8 mg/kg), via intravenous infusion every 4 weeks, during 4 phase III clinical trials. Serum concentration-time profiles of tocilizumab were adequately described by a 2-compartment disposition model with parallel linear and nonlinear elimination kinetics. The 8-mg/kg dose of tocilizumab, compared with the 4-mg/kg dose, resulted in a more pronounced saturation of the nonlinear clearance pathway over the dosing interval, and this nonlinear clearance was representative of target-mediated elimination due to tocilizumab binding to the IL-6 receptor.


Plasma protein-based signature predicts distant metastasis and induction chemotherapy benefit in Nasopharyngeal Carcinoma.
Rationale: Currently, for locoregionally advanced nasopharyngeal carcinoma (LA-NPC), there is no effective blood-based method to predict distant metastasis. We aimed to detect plasma protein profiles to identify biomarkers that could distinguish patients with NPC who are at high risk of posttreatment distant metastasis. Methods: A high-throughput antibody array was initially applied to detect 1000 proteins in pretreatment plasma from 16 matched LA-NPC patients with or without distant metastasis after radical treatment. Differentially expressed proteins were further examined using a low-throughput array to construct a plasma protein-based signature for distant metastasis (PSDM) in a cohort of 226 patients. Results: Fifty circulating proteins were differentially expressed between metastatic and non-metastatic patients and 18 were proven to be strongly correlated with distant metastasis-free survival (DMFS) in NPC. A PSDM signature consisting of five proteins (SLAMF5, ESM-1, MMP-8, INSR, and Serpin A5) was established to assign patients with NPC into a high-risk group and a low-risk group. Patients in the high-risk group had shorter DMFS (P < 0.001), disease-free survival (DFS) (P < 0.001) and overall survival (OS) (P < 0.001). Moreover, the PSDM performed better than N stage and Epstein-Barr virus (EBV) DNA load at effectively identifying patients with NPC at high risk of metastasis. For patients in the high-risk group, induction chemotherapy significantly improved DMFS, DFS, and OS. Conclusions: The PSDM could be a useful liquid biopsy tool to effectively predict distant metastasis and the benefit of induction chemotherapy in patients with LA-NPC.


[Overexpression of lncRNA MEG3 inhibits proliferation and invasion of glioblastoma U251 cells in vitro by suppressing HIF1α expression].
To investigate the effects of overexpression of long noncoding RNA (lncRNA) MEG3 on the proliferation and invasion of glioblastoma U251 cells by suppressing the expression of hypoxia inducible factor 1α(HIF1α). The expression of lncRNA MEG3 and HIF1α mRNA were examined in human fetal glial cells (HFGCs) and U251 cells using realtime quantitative PCR (qRT-PCR), and the expression of HIF1α protein was detected with Western blotting.U251 cells in normal culture or transfected with pcDNA3.1 vector (NC group) or pcDNA3.1-MEG3 vector via lipofectamine2000 were exposed to hypoxia for 12h, and the expressions of HIF1α mRNA and protein were detected with qRT-PCR and Western blotting, respectively.MTT assay and Transwell assay were employed to examine the influence of MEG3 overexpression on the proliferation and invasion of U251 cells. The expression of MEG3 was significantly lower and HIF1α mRNA and protein expressions were significantly higher in U251 cells than in HFGCs (P < 0.05).In U251 cells, overexpression of MEG3 significantly decreased the mRNA and protein expressions of HIF1α(P < 0.05).Hypoxic exposure for 12h also resulted in significantly lowered expression of HIF1α protein in U251 cells (P < 0.05).Overexpression of MEG3 obviously suppressed the proliferation and invasiveness of U251 cells (P < 0.05). MEG3 overexpression inhibits the proliferation and invasion of U251 cells through suppressing the expression of HIF1α mRNA and protein, suggesting that MEG3 may serve as a potential therapeutic target for glioblastomas.


Regulation of Plastid Gene Expression during Photooxidative Stress.
We have used the carotenoid biosynthesis inhibitor norflurazon to study the relationship between chloroplast and nuclear gene expression and the mechanisms by which plastid mRNA accumulation is regulated in response to photooxidative stress. By treating 4-week-old hydroponic spinach plants (Spinacea oleracea), we were able to determine the response at two distinct stages of chloroplast development. For all parameters studied, differences were found between the norflurazon-treated young and mature leaves. Young leaves lost essentially all pigment content in the presence of norflurazon, whereas mature leaves retained more than 60% of their chlorophyll and carotenoids. The accumulation of plastid mRNA was determined for several genes, and we found a decrease in mRNA levels for all genes except psbA in herbicide-treated young leaves. For genes such as atpB, psbB, and psaA, there was a corresponding change in the relative level of transcription, but for psbA and rbcL, transcription and mRNA accumulation were uncoupled. In norflurazon-treated mature leaves, all plastid mRNAs except psaA accumulated to normal levels, and transcription levels were either normal or higher than corresponding controls. This led to the conclusion that plastid mRNA accumulation is regulated both transcriptionally and posttranscriptionally in response to photooxidative stress. Although direct photooxidative damage is confined to the plastid and peroxisome, there is a feedback of information controlling the transcription of nuclear-encoded plastid proteins. Considerable evidence has accumulated implicating a "plastid factor" in this control. Therefore, the expression of several nuclear-encoded plastid proteins and the corresponding mRNAs were determined. Although the levels of both the small subunit of ribulose-1,5-bisphosphate carboxylase and the light harvesting chlorophyll a/b-binding protein and corresponding mRNAs were reduced, a 28-kilodalton chloroplast RNA-binding protein and corresponding mRNA were at normal levels in norflurazon-treated plants. Changes in mRNA and protein levels were not the result of a general loss due to photooxidation but rather the result of selective stabilization of certain components. The response of both genomes to photooxidative stress is discussed in terms of the postulated plastid factor.


Burns treatment in ancient times.
Discovery of fire at the dawn of prehistoric time brought not only the benefits to human beings offering the light and heat, but also misfortune due to burns; and that was the beginning of burns treatment. Egyptian doctors made medicines from plants, animal products and minerals, which they combined with magic and religious procedures. The earliest records described burns dressings with milk from mothers of male babies. Goddess Isis was called upon to help. Some remedies and procedures proved so successful that their application continued for centuries. The Edwin Smith papyrus (1500 BC) mentioned the treatment of burns with honey and grease. Ebers Papyrus (1500 BC) contains descriptions of application of mud, excrement, oil and plant extracts. They also used honey, Aloe and tannic acid to heal burns. Ancient Egyptians did not know about microorganisms but they knew that honey, moldy bread and copper salts could prevent infections from dirt in burns healing. Thyme, opium and belladona were used for pain relief. In the 4th century BC, Hippocrates recorded that Greek and Roman doctors used rendered pig fat, resin and bitumen to treat burns. Mixture of honey and bran, or lotion of wine and myrrh were used by Celsus. Honey was also known in Ayurveda (Indian medicine) time. Ayurvedic records Characa and Sushruta included honey in their dressing aids to purify sores and promote the healing. Burn treatment in Chinese medicine was traditional. It was a compilation of philosophy, knowledge and herbal medicine. The successful treatment of burns started in recent time and it has been made possible by better knowledge of the pathophysiology of thermal injuries and their consequences, medical technology advances and improved surgical techniques.


Why are older Floridians exceptional?
Why do older Floridians have higher utilization of health care and live longer than other older Americans? Higher health care use among Florida's older residents is likely related to housing patterns, marital status, health insurance coverage, and ethnic composition. Lower mortality is unlikely a result of lifestyle factors or labor-force participation rates but may be associated with usage of preventive health care services. These indicators suggest that attention needs to be paid to health behavior and social support networks.


Rural settlements transition (RST) in a suburban area of metropolis: Internal structure perspectives.
Rural settlements transition (RST) is one of the most significant indices for understanding the phenomena of rural reconstruction and urban-rural transformation in China. However, a systematic overview of RST is missing, and there is a lack of evidence regarding its characteristics from the internal structure perspectives. In this paper, we systematically explore the RST regarding spatio-temporal change characteristics of internal structure, patterns and impacts on rural environment and development by using practical survey internal land-use data from 2005 to 2015. The results show that the temporal change characteristics of the internal structure of rural settlements demonstrate a tendency for housing land to decrease and other land-use types to increase. The spatial change characteristics reveal that the structure inclines to more complexity and diversity from an exurban area to an urban-rural fringe area. Based on this finding, we identify that rapid development of rural industrialization, more agglomerate and effective industrial land-use, and improved public infrastructure construction are the general RST patterns. Spatially, there exists a physical decay pattern in the exurban area, thereby resulting in the hollowing-out of rural industries and of the population. In addition, the extensive and disorderly pattern in the suburban area causes low efficiency output and serious environmental pollution. The RST pattern in the urban hinterland promoted the "men-environment" compatible development. The study concludes that regional differentiation in patterns and impacts are significant in the process of RST. Future adaptive strategies for rural settlements adjustment should be conducted according to regional characteristics, including socio-economic status, physical geography condition and economic location to improve the rural environmental sustainability.


Macrophage proliferation, provenance, and plasticity in macroparasite infection.
Macrophages have long been center stage in the host response to microbial infection, but only in the past 10-15 years has there been a growing appreciation for their role in helminth infection and the associated type 2 response. Through the actions of the IL-4 receptor α (IL-4Rα), type 2 cytokines result in the accumulation of macrophages with a distinctive activation phenotype. Although our knowledge of IL-4Rα-induced genes is growing rapidly, the specific functions of these macrophages have yet to be established in most disease settings. Understanding the interplay between IL-4Rα-activated macrophages and the other cellular players is confounded by the enormous transcriptional heterogeneity within the macrophage population and by their highly plastic nature. Another level of complexity is added by the new knowledge that tissue macrophages can be derived either from a resident prenatal population or from blood monocyte recruitment and that IL-4 can increase macrophage numbers through proliferative expansion. Here, we review current knowledge on the contribution of macrophages to helminth killing and wound repair, with specific attention paid to distinct cellular origins and plasticity potential.


Temporally integrated single cell RNA sequencing analysis of PBMC from experimental and natural primary human DENV-1 infections.
Dengue human infection studies present an opportunity to address many longstanding questions in the field of flavivirus biology. However, limited data are available on how the immunological and transcriptional response elicited by an attenuated challenge virus compares to that associated with a wild-type DENV infection. To determine the kinetic transcriptional signature associated with experimental primary DENV-1 infection and to assess how closely this profile correlates with the transcriptional signature accompanying natural primary DENV-1 infection, we utilized scRNAseq to analyze PBMC from individuals enrolled in a DENV-1 human challenge study and from individuals experiencing a natural primary DENV-1 infection. While both experimental and natural primary DENV-1 infection resulted in overlapping patterns of inflammatory gene upregulation, natural primary DENV-1 infection was accompanied with a more pronounced suppression in gene products associated with protein translation and mitochondrial function, principally in monocytes. This suggests that the immune response elicited by experimental and natural primary DENV infection are similar, but that natural primary DENV-1 infection has a more pronounced impact on basic cellular processes to induce a multi-layered anti-viral state.


Constituents of the moss Polytrichum commune.
Two unusual flavonones coupled with styryl units, communins A (1) and B (2), and a new benzonaphthoxanthenone, ohioensin H (3), together with 11 known compounds, were isolated from the moss Polytrichum commune. The structures of 1-3 were assigned by spectroscopic data interpretation. The new compounds were evaluated for cytotoxicity against a small panel of cancer cell lines.


Expectation of life at birth, for India and major states, 1976-80.
Data are presented on life expectancy by sex for India and its constituent states for 1976-1980 using data from the Sample Registration System. "There is a wide variation among states in the expectation of life at birth. Uttar Pradesh has the lowest and Kerala has the highest expectation of life at birth. Within a state, the expectation of life at birth is higher for urban areas than for rural areas. Also the male expectation of life at birth is generally higher than female expectation in rural areas. However, the pattern is reversed in urban areas."