A surface-focused biotinylation procedure identifies the Yersinia pestis catalase KatY as a membrane-associated but non-surface-located protein.
This study identified major surface proteins of the plague bacterium Yersinia pestis. We applied a novel surface biotinylation method, followed by NeutrAvidin (NA) bead capture, on-bead digestion, and identification by liquid chromatography-tandem mass spectrometry (LC-MS-MS). The use of stachyose during biotinylation focused the reaction to the surface. Coupled with NA pulldown and immunoblot analysis, this method determined whether a protein was accessible to the surface. We applied the method to test the hypothesis that the catalase KatY is a surface protein of the plague bacterium Y. pestis. A rabbit serum recognized the catalase KatY as a major putative outer membrane-associated antigen expressed by Y. pestis cells grown at 37 degrees C. Similar findings by other groups had led to speculations that this protein might be exposed to the surface and might be a candidate for evaluation as a protective antigen for an improved plague vaccine. KatY was obtained only in the total membrane fraction, and stachyose greatly reduced its biotinylation as well as that of the periplasmic maltose binding protein, indicating that KatY is not on the bacterial surface. LC-MS-MS analysis of on-bead digests representing ca. 10(9) cells identified highly abundant species, including KatY, Pal, and OmpA, as well as the lipoprotein Pcp, all of which bound in a biotin-specific manner. Pla, Lpp, and OmpX (Ail) bound to the NA beads in a non-biotin-specific manner. There was no contamination from abundant cytoplasmic proteins. We hypothesize that OmpX and Pcp are highly abundant and likely to be important for the Y. pestis pathogenic process. We speculate that a portion of KatY associates with the outer membrane in intact cells but that it is located on the periplasmic side. Consistent with this idea, it did not protect C57BL/6 mice against bubonic plague.


Molecular characterization of newly recognized rodent parvoviruses.
Several autonomous rodent parvoviruses distinct from the prototypic rodent parvoviruses have been isolated. These include variants of a mouse parvovirus (MPV), a hamster isolate designated hamster parvovirus (HaPV), and a variant strain of minute virus of mice (MVM) designated MVM-Cutter or MVM(c). In this study, the DNA sequence of the coding regions of the viral genome and the predicted protein sequences for each of these new isolates were determined and compared to the immunosuppressive and prototypic strains of MVM [MVM(i) and MVM(p)], the rodent parvovirus H-1, and LuIII, an autonomous parvovirus of uncertain host origin. Sequence comparisons showed that the MPV isolates were almost identical, HaPV was very similar to MPV, and MVM(c) was most similar to MVM(i) and MVM(p). Haemagglutination inhibition assays revealed that MPV and HaPV represent two serotypes distinct from previously characterized rodent parvovirus serotypes while MVM(c) belongs to the MVM serotype. Each of the newly isolated rodent parvoviruses was shown to encapsidate a predominantly negative-sense 5 kb DNA genome and to encode two nonstructural proteins (NS1 and NS2) and two structural viral proteins (VP1 and VP2). These studies indicate that MPV and HaPV are autonomous parvoviruses distinct from previously characterized parvoviruses and MVM(c) is a variant strain of MVM distinct from MVM(i) and MVM(p).


Mass spectrometry imaging of thin-layer chromatography plates using laser desorption/low-temperature plasma ionisation.
Thin-layer chromatography (TLC) is a classic method for the separation and analysis of complex mixtures. Biological assays, chemical derivatisation and spectroscopy techniques are compatible with TLC and provide extra information about isolated compounds. However, coupling TLC to mass spectrometry is hampered by the difficulty to desorb the analytes from the silica surfaces. In this study, we used a multimodal ion source for laser desorption (LD) and low-temperature plasma (LTP) post-ionisation. Efficient desorption was reached by covering the TLC plates with activated carbon. Regions of interest can be analysed by spots, by lines or by area. We show the separation of methylxanthines from coffee, tea and cocoa preparations by TLC, with subsequent mass spectrometry imaging (MSI). Using a lateral resolution of 400 μm × 400 μm, allowed the acquisition of 21 895 spectra in 2.4 h (2.5 pixels per s). Further, we demonstrate the possibility of direct mass fragmentation studies and quantification. We mounted the system on an Open LabBot with a theoretical lateral resolution of 12.5 μm and performed the visualisation of ions of interest and the pixel-wise review of mass spectra with our free software RmsiGUI (). This non-proprietary and modular platform enables the cost-efficient adaption of the system and further development by the community.


Electronic cardiac medicine: present and future opportunities.
The second half of the 20th century witnessed a revolution in electronic medicine similar to that in pharmacology in the decades before. The advent of the implantable pacemaker, implantable cardioverter-defibrillators, cardiac resynchronisation therapies, insertable loop recorders and more, have improved diagnoses and reduced mortality and morbidity in millions of patients suffering from cardiac disease. The possibility to monitor patients continually without need for frequent office visits has the potential to reduce follow-up burden on physicians, facilitate increased use of home-based care and further improve the safety for patients. This review summarises the role of cardiac device therapies today and some of the developments which we can hope for in the nearest future.


Evaluation of knowledge base of hospital pharmacists and physicians on herbal medicines in Southwestern Nigeria.
The use of herbal medicines among Nigerian patients and the tendency to combine this class of medicines with allopathic drugs while on hospital admission have been on the increase. Earlier studies show that community pharmacists from Nigeria believe that they need more training to counsel patients on their use of herbal medications. To evaluate knowledge base, use, acceptability, attitudes and beliefs of hospital pharmacists and physicians in South-western Nigeria on herbal products / phytopharmaceuticals /dietary supplements (HP/PP/DS). A 20-odd questionnaire including an 8- item herbal medicine question was used to assess the knowledge of the physicians and pharmacists in the area of pharmacology of HP/PP/DS. Effects of demographic information such as age, sex, year post graduation, years of experience and area of specialization on the scores obtained were evaluated using Fisher's exact tests. Level of significance was set at p<0.05. Pharmacists (51.4%) believed that the training they had was not adequate while physicians (44.6%) believed HP/PP/DS are not safe and 18.5% believed they are not effective. Pharmacovigilance centres were not significantly used for adverse reactions reported by patients. Pharmacists (67.6%) and physicians (29.3%) believed phytopharmaceuticals interact with other drugs. Year of graduation had a significant effect on the perception of the possibilities of interaction for pharmacists. 5.6% of the physicians and 62.4% of the pharmacists scored ≥50% with the questions on use and interaction potentials of HP/PP/DS, with cadre and years of practice having significant effects. There is a deficiency in the knowledge base of physicians, especially on the pharmacology and potential interaction of herbal medicines. There is an urgent need for the inclusion of pharmacology of common herbal medicines in the curriculum of Nigerian medical degree and an improvement in the pharmacy curriculum in this area.


Taenia hydatigena cysticercosis in wild boar (Sus scrofa) from southern Italy: an epidemiological and molecular survey.
Taenia hydatigena cysticercosis is a widespread parasitic disease of wild and domestic animals. In Europe, the increase in wild boar population may potentially contribute to the spread of this parasitic infection. To determine the occurrence of cysticerci (metacestodes) in wild boar population from southern Italy, carcasses were inspected during three hunting seasons (2016-2018). Out of 3363 wild boar examined, 229 (6.8%) harboured cysticerci with 188 (82.1%) infected by a single cyst, vs 41 (17.9%) boars having more than one. Most of the positive animals (187; 81.7%) showed cysts on the liver, whereas a multiple localization of cysticerci was reported in 10 (4.4%) wild boar. The total number of cysts retrieved from positive animals was 301 (average 1.3). Molecular analysis revealed the occurrence of a common haplotype (Hap 8) shared between wild boar and domestic animals. Our findings suggest the presence of a T. hydatigena semi-domestic life cycle in which wild boar may play an important role, due to a large number of offal available to hunting dogs, wolves and foxes during hunting seasons. Hunters may be players in the management of wildlife species to control and prevent the circulation of parasitic diseases.


Plant-Produced Glycosylated and In Vivo Deglycosylated Receptor Binding Domain Proteins of SARS-CoV-2 Induce Potent Neutralizing Responses in Mice.
The COVID-19 pandemic, caused by SARS-CoV-2, has rapidly spread to more than 222 countries and has put global public health at high risk. The world urgently needs cost-effective and safe SARS-CoV-2 vaccines, antiviral, and therapeutic drugs to control it. In this study, we engineered the receptor binding domain (RBD) of the SARS-CoV-2 spike (S) protein and produced it in the plant Nicotiana benthamiana in a glycosylated and deglycosylated form. Expression levels of both glycosylated (gRBD) and deglycosylated (dRBD) RBD were greater than 45 mg/kg fresh weight. The purification yields were 22 mg of pure protein/kg of plant biomass for gRBD and 20 mg for dRBD, which would be sufficient for commercialization of these vaccine candidates. The purified plant-produced RBD protein was recognized by an S protein-specific monoclonal antibody, demonstrating specific reactivity of the antibody to the plant-produced RBD proteins. The SARS-CoV-2 RBD showed specific binding to angiotensin converting enzyme 2 (ACE2), the SARS-CoV-2 receptor. In mice, the plant-produced RBD antigens elicited high titers of antibodies with a potent virus-neutralizing activity. To our knowledge, this is the first report demonstrating that mice immunized with plant-produced deglycosylated RBD form elicited high titer of RBD-specific antibodies with potent neutralizing activity against SARS-CoV-2 infection. Thus, obtained data support that plant-produced glycosylated and in vivo deglycosylated RBD antigens, developed in this study, are promising vaccine candidates for the prevention of COVID-19.


DNA origami as a carrier for circumvention of drug resistance.
Although a multitude of promising anti-cancer drugs have been developed over the past 50 years, effective delivery of the drugs to diseased cells remains a challenge. Recently, nanoparticles have been used as drug delivery vehicles due to their high delivery efficiencies and the possibility to circumvent cellular drug resistance. However, the lack of biocompatibility and inability to engineer spatially addressable surfaces for multi-functional activity remains an obstacle to their widespread use. Here we present a novel drug carrier system based on self-assembled, spatially addressable DNA origami nanostructures that confronts these limitations. Doxorubicin, a well-known anti-cancer drug, was non-covalently attached to DNA origami nanostructures through intercalation. A high level of drug loading efficiency was achieved, and the complex exhibited prominent cytotoxicity not only to regular human breast adenocarcinoma cancer cells (MCF 7), but more importantly to doxorubicin-resistant cancer cells, inducing a remarkable reversal of phenotype resistance. With the DNA origami drug delivery vehicles, the cellular internalization of doxorubicin was increased, which contributed to the significant enhancement of cell-killing activity to doxorubicin-resistant MCF 7 cells. Presumably, the activity of doxorubicin-loaded DNA origami inhibits lysosomal acidification, resulting in cellular redistribution of the drug to action sites. Our results suggest that DNA origami has immense potential as an efficient, biocompatible drug carrier and delivery vehicle in the treatment of cancer.


Intelligent backtracking in plan-based deduction.
This paper develops a method of mechanical deduction based on a graphical representation of the structure of proofs. Attempts to find a refutation(s) are recorded in the form of plans, corresponding to portions of an AND/OR graph search space and representing a purely deductive structure of derivation. This method can be applied to any initial base (set of nonnecessarily Horn clauses). Unlike the exhaustive (blind) backtracking which treats all the goals deduced in the course of a proof as equally probable sources of failure, his approach detects the exact source of failure. Only a small fragment of the solution space is kept on disk as a collection of pairs, each of which consists of a plan and a graph of constraints. The search strategy and the method of nonredundant processing of individual pairs which leads to a solution (if it exists) is presented. This approach is compared¿on a special case¿with a blind backtracking algorithm for which an exponential improvement is demonstrated. Some important implementation problems are discussed, and toplevel design of a mechanical deduction system implementing our algorithm is presented. It is proven that the algorithm is complete in the following sense: if for a given base a resolution refutation exists, then this refutation is found by the algorithm.


Distinct amygdalar AMPAergic/GABAergic mechanisms promote anxiolitic-like effects in an unpredictable stress model of the hamster.
Studies have pointed to both α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor (AMPAR) antagonists and GABA(A) receptor (GABA(A)R) agonists as potent antistress agents. In this work, separate subchronic injections of the AMPAR antagonist, 6-ciano-7-nitroquinoxaline-2,3-dione (CNQX), and α1 GABA(A)R subunit agonist (Zol) within the central amygdala nucleus modified the elevated plus maze performances of hamsters exposed randomly to one of the following stressful conditions: food/water deprivation, forced swimming test, and permanence in cold room. Indeed, stressed hamsters treated with CNQX or Zol displayed a very great (p < 0.001) increase of entrance plus a moderate (p < 0.05) time spent into open arms, respectively. At the cellular level, Zol-treated animals supplied a moderately evident argyrophilic reaction (indicative of neurodegeneration) in the hippocampus while it was absent in the hypothalamus. Interestingly, this reaction was significantly reduced by CNQX supporting its preferential protective role. Furthermore, both agents were responsible for a mixed expression pattern of GluR1 and GluR2 mRNA levels in which Zol overall upregulated GluR1 mRNAs, while they were downregulated by CNQX in the hippocampal oriens-pyramidalis layer and in layer III of the cerebral cortex. These findings support the amygdalar AMPAergic protective response against anxiety states in chronically stressed hamsters, which may constitute useful therapeutic strategies for panic-related mood disorders.


Functional effects and gender association of COX-2 gene polymorphism G-765C in bronchial asthma.
Prostaglandins, generated via the COX pathways, are essential mediators of inflammation in bronchial asthma. The promoter polymorphism of COX-2 gene (G-765C), which might affect binding of transcription factors, has recently been described To study distribution and function of the genetic COX-2 variant in patients with asthma compared with healthy controls. Three groups of adults were studied: (1) patients with aspirin-induced asthma (AIA; n=112), (2) asthmatic patients who tolerated aspirin (ATA; n=198), and (3) a random population sample from city of Krakow (n=547). The COX-2 promoter region was genotyped for the G-765C polymorphism. Ex vivo production of prostaglandin E2 and prostaglandin D2 by peripheral blood monocytes was measured. In the 2 asthmatic groups, the G-765C allele frequency was similar (AIA, 0.18; ATA, 0.19) and did not differ from that of controls (0.17). In asthmatic women, but not in men, CC homozygotes were overrepresented compared with controls (odds ratio, 3.08; 95% CI, 1.35-6.63; P=.01). There was no relationship between genotype and FEV1, serum IgE, blood eosinophil count, or duration of the disease. In AIA but not in ATA patients, CC homozygosity was associated with more severe course of the disease, as reflected by need for oral corticotherapy. Production of 2 prostaglandins by monocytes was more than 10-fold higher in CC than in GG homozygotes, and the magnitude of this difference was not changed by LPS stimulation. In asthma, the COX-2 -765C homozygosity is associated with female sex. The CC homozygosity has functional effects resulting in increased capacity of monocytes to produce prostaglandins.


Role of HLA-DQ typing and antitissue transglutaminase antibody titres in diagnosing coeliac disease among Sudanese children with type 1 diabetes mellitus.
The aim of the study was to determine the prevalence of coeliac disease (CD) and to recognise Human leukocyte antigen (HLA)-associated hereditary susceptibility to Sudanese CD patients with type 1 diabetes mellitus (DM1). Antitissue transglutaminase IgA (anti-TG IgA) was measured in the serum of 373 children affected with DM1 aged 1-19-year old and in 100 serum samples from non-diabetic control children. Histological examination was performed in 19 children seropositive for anti-TG IgA (17 DMI and 2 controls). Additionally, PCR-based analysis of Major histocompatibility complex, class II, DQ beta 1 (HLA-DQB1) genotyping was implemented in three study population groups as follows: group 1 (n=25) (+ve DM1 and +ve CD), group 2 (n=63) (-ve DM1 and +ve CD) and control group 3 (n=2) (+ve CD). Twenty-six Sudanese children with DM1 out of 373 (6.97%) were seropositive for anti-TG IgA. Duodenal biopsy revealed Marsh 2 and 3 in 13 out of 17 (76.47%) seropositive anti-TG IgA patients with DM1. Significant association (p<0.05) was detected between the level of anti-TG IgA autoantibodies (IU/mL) and Marsh stage. HLA DQ2 and DQ8 were found in 88% (22/25) and 8% (2/25) of examined patients with CD with DM1, respectively. Anti-TG IgA titre of greater than 10 times upper limit of normal (≥10× ULN) can be useful for detecting CD in children with type 1 diabetes without duodenal biopsy. HLA testing in children with DM1 appears to provide little added benefit given the high prevalence (96%) of HLA DQ2/DQ8 in children with DM1.


Disalicylic Acid Provides Effective Control of Pectobacterium brasiliense.
Bis(2-carboxyphenyl) succinate (disalicylic acid; DSA) is composed of two salicylic acids connected by a succinyl linker. Here, we propose its use as a new, synthetic plant-protection agent. DSA was shown to control Pectobacterium brasiliense, an emerging soft-rot pathogen of potato and ornamental crops, at minimal inhibitory concentrations (MIC) lower than those of salicylic acid. Our computational-docking analysis predicted that DSA would inhibit the quorum-sensing (QS) synthase of P. brasiliense ExpI more strongly than SA would. In fact, applying DSA to P. brasiliense inhibited its biofilm formation, secretion of plant cell wall-degrading enzymes, motility and production of acyl-homoserine lactones (AHL) and, subsequently, impaired its virulence. DSA also inhibited the production of AHL by a QS-negative Escherichia coli strain (DH5α) that had been transformed with P. brasiliense AHL synthase, as demonstrated by the biosensors Chromobacterium violaceaum CV026 and E. coli pSB401. Inhibition of the QS machinery appears to be one of the mechanisms by which DSA inhibits specific virulence determinants. A new route is proposed for the synthesis of DSA, which holds greater potential for use as an anti-virulence agent than its precursor SA. Based on these findings, DSA is an excellent candidate for repurposing for new applications.


MTHFR 677 CT/MTHFR 1298 CC genotypes are associated with increased risk of hypertension in Indians.
The goals of our present study were to measure plasma homocysteine levels and determine their association with methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms (C677T and A1298C) in essential hypertensive subjects. Plasma total homocysteine and folic acid levels were measured in essential hypertensive patients (n = 153) before and after oral supplementation with either 5 mg folic acid tablet/day or 5 mg placebo/day for 4 weeks and compared with age and sex matched normotensive controls (n = 133). MTHFR gene polymorphisms (C677T and A1298C) were studied by restriction fragment length polymorphism and correlated with plasma homocysteine levels. Homocysteine levels were significantly higher in hypertensive patients as compared to controls and showed a negative correlation with plasma folate levels. Folic acid supplementation (5 mg/day) for 4 weeks resulted in a significant decrease in plasma homocysteine concentrations in these patients. Patients carrying MTHFR 677T allele (OR = 1.90; 95%CI: 1.14-3.19) or MTHFR 1298C (OR = 2.6, 95%CI: 1.55-4.40) allele were at increased risk of hypertension. The frequency of co-occurrence of MTHFR 677 CT/1298 CC genotypes was significantly higher in the patients compared to controls (P < 0.05) and was associated with increased risk of hypertension (OR = 3.54, 95%CI: 0.37-4.30). Subjects with MTHFR 1298 CC genotype had significantly higher homocysteine levels compared to those with MTHFR 1298 AA genotype (P < 0.05). Our results indicate that MTHFR 677T and 1298C alleles and co-occurrence of MTHFR 677 CT/MTHFR 1298 CC genotypes are associated with increased risk of hypertension and MTHFR 1298 CC genotype is associated with higher homocysteine levels in our subjects.


Identifying Typologies of Persons Who Died by Suicide: Characterizing Suicide in Victoria, Australia.
The objective of the study was to determine whether people who died by suicide form groups based on demographic, psychosocial, mental and physical health factors and exposure to stressors. A retrospective case series review of 2,839 individuals who died by suicide in Victoria, Australia over the period 2009-2013 was conducted. A two-stage cluster analysis was performed. Diagnosis of mental illness was present in 52% of cases and initial cluster analysis determined two groups with the main predictor of group membership being the presence of diagnosed mental illness. Further analysis identified four subgroups within the mental illness group and two within the non-mental illness group. The study demonstrates that suicide does not always occur in the context of mental illness; people who die by suicide cannot be considered a homogenous group.


Creating Gameful Design in mHealth: A Participatory Co-Design Approach.
Gameful designs (gamification), using design pieces and concepts typically found in the world of games, is a promising approach to increase users' engagement with, and adherence to, electronic health and mobile health (mHealth) tools. Even though both identifying and addressing users' requirements and needs are important steps of designing information technology tools, little is known about the users' requirements and preferences for gameful designs in the context of self-management of chronic conditions. This study aimed to present findings as well as the applied methods and design activities from a series of participatory design workshops with patients with chronic conditions, organized to generate and explore user needs, preferences, and ideas to the implementation of gameful designs in an mHealth self-management app. We conducted three sets of two consecutive co-design workshops with a total of 22 participants with chronic conditions. In the workshops, we applied participatory design methods to engage users in different activities such as design games, scenario making, prototyping, and sticky notes exercises. The workshops were filmed, and the participants' interactions, written products, ideas, and suggestions were analyzed thematically. During the workshops, the participants identified a wide range of requirements, concerns, and ideas for using the gameful elements in the design of an mHealth self-management app. Overall inputs on the design of the app concerned aspects such as providing a positive user experience by promoting collaboration and not visibly losing to someone or by designing all feedback in the app to be uplifting and positive. The participants provided both general inputs (regarding the degree of competitiveness, use of rewards, or possibilities for customization) and specific inputs (such as being able to customize the look of their avatars or by having rewards that can be exchanged for real-world goods in a gift shop). However, inputs also highlighted the importance of making tools that provide features that are meaningful and motivating on their own and do not only have to rely on gameful design features to make people use them. The main contribution in this study was users' contextualized and richly described needs and requirements for gamefully designed mHealth tools for supporting chronic patients in self-management as well as the methods and techniques used to facilitate and support both the participant's creativity and communication of ideas and inputs. The range, variety, and depth of the inputs from our participants also showed the appropriateness of our design approach and activities. These findings may be combined with literature and relevant theories to further inform in the selection and application of gameful designs in mHealth apps, or they can be used as a starting point for conducting more participatory workshops focused on co-designing gameful health apps.


DJ-1, PINK1, and their effects on mitochondrial pathways.
Genetic forms of parkinsonism are interesting for two particular reasons. First, finding a gene identifies a cause for a disease that would otherwise be unexplained. Second, finding several genes for the same disorder allows us to reconstruct molecular pathways that, in the example of Parkinson's disease, are be associated with the survival of dopamine neurons in the substantia nigra. Two rare causes of parkinsonism, DJ-1 and PINK1, are associated with mitochondria. This organelle has long been linked with Parkinson's disease, and recent results are starting to show how mutations impact mitochondrial function. In this short review, I will discuss how we can use some of this information to understand why it is that neurons become dysfunctional in PD.


A density functional theory study for the hydrogen-bonded nucleic acid base pair: cytosine dimer.
Theoretical investigation for the geometric and energetic properties, rotational constants, harmonic vibrational frequencies, and binding energies of nucleic acid base pair, cytosine dimer, are carried out by using the density functional theory method. The dimer structures resulting from both the keto and the enol (cis/trans) tautomers are investigated in the present study. Various isomers are considered to find the stable structures of the cytosine dimer. The planar cytosine dimer, K-K3 with C2h symmetry, resulting from nonplanar keto tautomers, is found to be thermodynamically most stable out of the four different stable isomers and having the highest binding energy value, 19.51 kcal/mol (including basis set superposition error correction). The vibrational frequency analysis also suggests a red shift of 367.97 cm(-1) for the hydrogen-bonding K-K3 symmetric dimer with two hydrogen bond lengths, each of length 1.913 angstroms. Moreover, charge distribution (ChelpG charges), Laplacian electronic density distribution, and the dimerization equilibrium for the most stable dimer, K-K3, have also been investigated using the same method and the basis set.


Effect of methylprednisolone and of ibuprofen, a nonsteroidal antiinflammatory agent, on bronchoalveolar inflammation following endotoxemia.
Septicemia by gram-negative organisms is a common cause of the adult respiratory distress syndrome (ARDS). The role of neutrophils in causing parenchymal lung damage in ARDS has recently been emphasized. A single intraperitoneal injection of Escherichia coli endotoxin in rats causes acute neutrophil alveolitis similar to that of ARDS. We studied the ability of pretreatment with either ibuprofen (IBU) or methylprednisolone (MP) to ablate directly the alveolar inflammatory response to endotoxin in the rat model. To compare the severity of inflammation, we quantified inflammatory cell recovery by whole lung lavage 24 hours after injection of endotoxin, the time point at which neutrophil alveolitis due to endotoxin is most intense. Pretreatment with a single dose of IBU 3.75 mg/kg prior to endotoxin injections was associated with a significant increase in the total number of inflammatory cells, and in both the percentage and the absolute number of neutrophils recovered from the lung, despite significantly decreasing the plasma level of thromboxane B2, which increased 10-fold after endotoxin. Paradoxically, IBU 30 mg/kg significantly decreased the intensity of neutrophil alveolitis. MP 30 mg/kg had no effect on recovery of inflammatory cells from the lung by bronchoalveolar lavage following endotoxin. Cyclooxygenase inhibitors such as ibuprofen may cause a dose-dependent biphasic effect on lung inflammation following endotoxin: enhancement of inflammation at a low dose and suppression of inflammation at a high dose.


Simultaneous Endocardial and Epicardial Delineation of 3D Reentrant Ventricular Tachycardia.
Mechanisms of scar-related ventricular tachycardia (VT) are largely based on computational and animal models that portray a 2-dimensional view. The authors sought to delineate the human VT circuit with a 3-dimensional perspective from recordings obtained by simultaneous endocardial and epicardial mapping. High-resolution mapping was performed during 97 procedures in 89 patients with structural heart disease. Circuits were characterized by systematic isochronal analysis to estimate the dimensions of the isthmus and extent of the exit region recorded on both myocardial surfaces. A total of 151 VT morphologies were mapped, of which 83 underwent simultaneous endocardial and epicardial mapping; 17% of circuits activated in a 2-dimensional plane, restricted to 1 myocardial surface. Three-dimensional activation patterns with nonuniform transmural propagation were observed in 61% of circuits with only 4% showing transmurally uniform activation, and 18% exhibiting focal activation patterns consistent with mid-myocardial reentry. The dimensions of the central isthmus were 17 mm (12 to 28 mm) × 10 mm (9 to 19 mm) with 55% exhibiting a minimal dimension of <1.5 cm. QRS activation was transmural in 63% and located 43 mm (34 to 52 mm) from the central isthmus. On the basis of 6 proposed definitions for epicardial VT, the prevalence of an epicardial circuit ranged from 21% to 80% in ischemic cardiomyopathy and 28% to 77% in nonischemic cardiomyopathy. A 2D perspective oversimplifies the electrophysiological circuit responsible for reentrant human VT and simultaneous endocardial and epicardial mapping facilitates inferences about mid-myocardial activation. Intricate activation patterns are frequently observed on both myocardial surfaces, and the epicardium is functionally involved in the majority of circuits. Human reentry may exist within isthmus dimensions smaller than 1 cm, whereas QRS activation is often transmural and remote from the critical isthmus target. A 3-dimensional perspective of the VT circuit may enhance the precision of ablative therapy and may support a greater role for adjunctive strategies and technology to address arrhythmogenic tissue harbored in the mid-myocardium and subepicardium.


Pathophysiology of ischemic cell death: II. Changes in plasma membrane permeability and cell volume.
Isolated rabbit retinas were subjected for various durations to several types of ischemic insult, and then returned to control medium for periods of up to 4 3/4 h before measurements were made of total water, inulin-free water, and plasma membrane permeability as assessed by mannitol penetration into the inulin-free water. Neither anoxia nor substrate deprivation alone, for as long as 50 min, caused significant irreversible swelling, but they were synergistic in combination. Restricting the volume of extracellular fluid during the combined deprivation caused the changes responsible for swelling to occur much sooner. There was a progressive increase in membrane permeability, with a delayed increase in intracellular water beginning about 2 h after the ischemic insult. Cell swelling correlated closely with loss of viability as evidenced by failure to reinstitute protein synthesis, but the swelling appeared to be the consequence rather than the cause of the initial irreversible damage.


Z-scheme inverse opal CN/BiOBr photocatalysts for highly efficient degradation of antibiotics.
Optimizing the heterojunction structure of semiconductor photocatalysts is vital for utilizing their abilities in organic matter degradation. Herein, a novel fabrication of a Z-scheme system with inverse opal g-C3N4 and BiOBr via a reflux process is developed. On the one hand, the unique inverse opal construction, formed by using silica (SiO2) photonic crystals as the template, not only provides a larger surface area for adsorption of antibiotics but also improves the separation and transfer efficiency of photogenerated electrons and holes as well as visible light absorption ability. On the other hand, by imitating natural photosynthesis, the artificial Z-scheme system with an inverse opal g-C3N4-BiOBr nanojunction further promotes the separation of photogenerated electrons and holes. The obtained IO CN/BiOBr catalyst exhibits superior photocatalytic performance for antibiotics degradation under visible light irradiation. In a typical test, almost complete degradation of levofloxacin (LVX) (10 mg L-1) could be achieved within 50 min due to the proper bandgap match between inverse opal g-C3N4 and BiOBr as well as enhanced surface area and light absorption. The present work provides an alternative strategy for construction of novel visible-light-driven Z-scheme photocatalysts and offers some new insights into the methods of waste water treatment.


A comparative study of zoledronic acid and once weekly Alendronate in the management of acute Charcot arthropathy of foot in patients with diabetes mellitus.
The aim of this study was to assess and compare the response to two forms of treatment-immobilization with zoledronic acid injection and immobilization with oral weekly Alendronate, in patients with diabetes mellitus and acute Charcot arthropathy (CA) of foot in terms of clinical and radiological parameters. Patients attending the endocrinology and podiatry clinic with history of diabetes mellitus and Acute CA were taken for study. The patients were randomized into two treatment groups. Group Z-zoledronic acid injection along with total contact cast (TCC). Group A-Tab. Alendronate 70 mg. once a week till the complete clinical resolution of acute CA along with TCC. Forty-five patients were randomized and 40 of them completed the study. The primary end point was complete clinical resolution of acute CA-defined as temperature difference between normal and affected foot <1°F. Among the 40 patients, 30 (75%) had complete clinical resolution. The mean number of days taken for complete clinical resolution since the initiation of treatment (either Zoledronic acid or Alendronate) was approximately 122 days. There was no significant difference in a number of days required for complete clinical resolution, between the two forms of therapy. There was more than 50% reduction in the visual score between the baseline and the final scan. The target to non-target ratio in the skeletal phase also showed an average of 40% reduction from the baseline to the final skeletal scintigraphy. Both Intravenous Zoledronic acid and oral alendronate had comparable efficacy with respect to the time taken for attaining complete clinical resolution of acute CA of foot. However, Alendronate therapy was cost effective among the two. (99m)Tc MDP bone scan can be used as an adjuvant to the clinical parameters in assessing the response to therapy.


Neural responses to emotional expression information in high- and low-spatial frequency in autism: evidence for a cortical dysfunction.
Despite an overall consensus that Autism Spectrum Disorder (ASD) entails atypical processing of human faces and emotional expressions, the role of neural structures involved in early facial processing remains unresolved. An influential model for the neurotypical brain suggests that face processing in the fusiform gyrus and the amygdala is based on both high-spatial frequency (HSF) information carried by a parvocellular pathway, and low-spatial frequency (LSF) information separately conveyed by a magnocellular pathway. Here, we tested the fusiform gyrus and amygdala sensitivity to emotional face information conveyed by these distinct pathways in ASD individuals (and matched Controls). During functional Magnetical Resonance Imaging (fMRI), participants reported the apparent gender of hybrid face stimuli, made by merging two different faces (one in LSF and the other in HSF), out of which one displayed an emotional expression (fearful or happy) and the other was neutral. Controls exhibited increased fusiform activity to hybrid faces with an emotional expression (relative to hybrids composed only with neutral faces), regardless of whether this was conveyed by LSFs or HSFs in hybrid stimuli. ASD individuals showed intact fusiform response to LSF, but not HSF, expressions. Furthermore, the amygdala (and the ventral occipital cortex) was more sensitive to HSF than LSF expressions in Controls, but exhibited an opposite preference in ASD. Our data suggest spared LSF face processing in ASD, while cortical analysis of HSF expression cues appears affected. These findings converge with recent accounts suggesting that ASD might be characterized by a difficulty in integrating multiple local information and cause global processing troubles unexplained by losses in low spatial frequency inputs.


Effect of complexed trace minerals on cumulus-oocyte complex recovery and in vitro embryo production in beef cattle1,2.
The objective of this experiment was to evaluate the impact of complexed trace mineral supplementation on ovum pick-up (OPU) and in vitro embryo production in lactating beef cows. Thirty days prior to fixed-time artificial insemination (FTAI; day -30), 68 postpartum cows were stratified by BW, BCS, and parity before being randomly assigned to 10 pens of either a treatment (TRT; n = 5) or a control (CNT; n = 5) group. Each group received a weekly mineral supplement allotment of 1.16 kg × week-1 × cow-calf pair-1 for 14 wk. Cows assigned to the TRT group received a mineral supplement that contained amino acid complexes of zinc, copper, and manganese, as well as cobalt glucoheptonate (Availa Plus; Zinpro Corp., Eden Prairie, MN, USA), while cows assigned to the CNT group received a mineral supplement that was formulated to contain similar concentrations of these trace minerals from inorganic sources. All cows were submitted to a 7 d CO-Synch + CIDR protocol on day -10 and bred using FTAI on day 0. Pregnancy diagnosis was performed on day 28 and nonpregnant cows were removed. All pregnant cows were subjected to ovum pick-up (OPU) on day 52 and 67 of gestation. Cumulus-oocyte complexes (COCs) were evaluated and graded prior to in vitro fertilization (IVF). Analysis of variance was conducted to determine effects of treatment on response variables, and pen was considered the experimental unit. Supplement consumption did not differ (P = 0.48) between treatments (1.16 ± 0.12 vs. 1.07 ± 0.15 kg of DM × week-1 × cow-calf pair-1 for TRT and CNT, respectively). Total COC recovery was greater (P = 0.03) from TRT when compared with CNT cows (22.4 ± 2.0 vs. 16.4 ± 1.4 COCs × pen-1, respectively) and the number of COCs meeting maturation criteria was increased in TRT cows (P = 0.05) when compared with CNT cows (15.9 ± 1.6 vs. 11.8 ± 1.0 COCs × pen-1, respectively). Production of transferable embryos tended to be greater (P = 0.06) for TRT than CNT cows (4.7 ± 0.6 vs. 2.7 ± 0.7 embryos × pen-1, respectively). Furthermore, when expressed as a ratio, the number of recovered COCs meeting maturation criteria that were required to produce a transferable embryo tended to be lower for TRT than CNT cows (3.10 ± 0.93 vs. 7.02 ± 1.60; P = 0.06). In summary, complete replacement with complexed trace mineral improved COC recovery and in vitro embryo production when compared with inorganic forms of these trace minerals in beef cows.


[Treatment of perforated gastroduodenal ulcers].
139 patients with perforated ulcers of the stomach and the duodenum were operated. Diffuse peritonitis was encountered in 117 (84.3%) patients. Radical organ-saving operations were carried out in 111 (79.9%) patients. 6 patients (4.3%) died after operation. The authors suggest that usage of organ-saving operations in perforated pyloroduodenal ulcers complicated by peritonitis is quite justified and does not lead to increase in postoperative complications and mortality rate.


Dictyostelium mutants lacking multiple classic myosin I isoforms reveal combinations of shared and distinct functions.
Dictyostelium cells that lack the myoB isoform were previously shown to exhibit reduced efficiencies of phagocytosis and chemotactic aggregation ("streaming") and to crawl at about half the speed of wild-type cells. Of the four other Dictyostelium myosin I isoforms identified to date, myoC and myoD are the most similar to myoB in terms of tail domain sequence. Furthermore, we show here that myoC, like myoB and myoD, is concentrated in actin-rich cortical regions like the leading edge of migrating cells. To look for evidence of functional overlap between these isoforms, we analyzed myoB, myoC, and myoD single mutants, myoB/myoD double mutants, and myoB/myoC/myoD triple mutants, which were created using a combination of gene targeting techniques and constitutive expression of antisense RNA. With regard to the speed of locomoting, aggregation-stage cells, of the three single mutants, only the myoB mutant was significantly slower. Moreover, double and triple mutants were only slightly slower than the myoB single mutant. Consistent with this, the protein level of myoB alone rises dramatically during early development, suggesting that a special demand is placed on this one isoform when cells become highly motile. We also found, however, that the absolute amount of myoB protein in aggregation-stage cells is much higher than that for myoC and myoD, suggesting that what appears to be a case of nonoverlapping function could be the result of large differences in the amounts of functionally overlapping isoforms. Streaming assays also suggest that myoC plays a significant role in some aspect of motility other than cell speed. With regard to phagocytosis, both myoB and myoC single mutants exhibited significant reductions in initial rate, suggesting that these two isoforms perform nonredundant roles in supporting the phagocytic process. In triple mutants these defects were not additive, however. Finally, because double and triple mutants exhibited significant and progressive decreases in doubling times, we also measured the kinetics of fluid phase endocytic flux (uptake, transit time, efflux). Not only do all three isoforms contribute to this process, but their contributions are synergistic. While these results, when taken together, refute the simple notion that these three "classic" myosin I isoforms perform exclusively identical functions, they do reveal that all three share in supporting at least one cellular process (endocytosis), and they identify several other processes (motility, streaming, and phagocytosis) that are supported to a significant extent by either individual isoforms or various combinations of them.


Comparison of Minimally Invasive Procedures for Benign Prostatic Hyperplasia: A Cost-effectiveness Analysis.
Background Benign prostatic hyperplasia (BPH) is a disease that affects millions of U.S. men and is costly to treat. Purpose To compare the cost-effectiveness of four minimally invasive therapies (MITs) and medical management for the treatment of BPH. Materials and Methods A cost-effectiveness analysis from a payer's perspective with Markov modeling was performed, comparing prostatic artery embolization (PAE), prostatic urethral lift, aquablation, water vapor thermal therapy, and medical management for BPH spanning a time horizon of 5 years. The model incorporated the probability of procedural complications and recurrent symptoms necessitating retreatment, which were extracted from published studies with long-term follow-up. Costs were based on Medicare reimbursements using CPT codes for ambulatory surgery centers. Outcomes were measured using the quality-adjusted life year (QALY), incorporating both life quality and expectancy. Statistical analyses included a base case calculation (using the most probable value of each parameter) and probabilistic and deterministic sensitivity analyses. Results In the base case calculation, outcomes for the strategies were comparable, with a difference of 0.030 QALY (11 days of life in perfect health) between the most (PAE) and least (medical management) effective strategies. PAE was the most cost-effective strategy relative to medical management, with an incremental cost-effectiveness ratio of $64 842 per QALY. Probabilistic sensitivity analysis showed PAE was more cost-effective compared with prostatic urethral lift, aquablation, water vapor therapy, and medical management in pairwise comparisons. In sensitivity analysis of retreatment risk, PAE remained the most cost-effective strategy until its repeat treatment rates exceeded 2.30% per 6 months, at which point water vapor therapy became the optimal choice. PAE was the most cost-effective procedure when its procedural cost was lower than $4755. Aquablation and prostatic urethral lift became more cost-effective when their procedural costs were lower than $3015 and $1097, respectively. Conclusion This modeling-based study showed that PAE appears to be a cost-effective modality among medical management and MITs for patients with BPH, with comparable outcomes to prostatic urethral lift, water vapor therapy, and aquablation at a lower expected cost. © RSNA, 2023 Supplemental material is available for this article. See also the editorial by Gemmete in this issue.


Dopamine D2 receptor gene (DRD2) is associated with alcoholism with conduct disorder.
This study examined whether there is evidence for an association between alcoholism with conduct disorder and alleles of the TaqI A and TaqI B polymorphisms, both individually and as haplotypes, at the dopamine D2 receptor gene (DRD2). We studied 182 Han Chinese subjects, including 34 alcoholics with conduct disorder, 63 alcoholics without conduct disorder, and 85 nonalcoholics. Alcohol dependence and conduct disorder were defined according to DSM-III-R criteria. Significant associations were observed between TaqI A and TaqI B at the DRD2 locus, tested individually and as haplotypes, and alcoholism with conduct disorder. Our results suggested that DRD2 might be associated with conduct disorder or a predisposition to both conduct disorder and alcoholism. However, this needs to be further investigated by examining the differences among conduct disorder with alcoholism, conduct disorder only, and controls for the TaqI A and B system at DRD2.


Prevalence trends of the new variant of Chlamydia trachomatis in four counties of Sweden in 2007-2011.
A new variant of Chlamydia trachomatis (nvCT) was discovered in Sweden in 2006, and it could not be detected by diagnostic systems from Abbott and Roche, whereas the third system used, from Becton Dickinson (BD), detects nvCT. We analyzed 3648 samples from 2 counties that used Roche and 2 counties that used BD methods from 2007 to 2011. After implementation of a Roche method that detects nvCT, its proportion has decreased and converged in the 4 counties but are still at different levels in Roche and BD counties. Future studies are needed to see if nvCT will decline further.


Inter-regional contribution of enhanced activity of the primary somatosensory cortex to the anterior cingulate cortex accelerates chronic pain behavior.
Multiple cortical areas are involved in pain processing, including the primary somatosensory cortex (S1) and the anterior cingulate cortex (ACC). Although accumulations of evidence suggest that the S1 activity increases under chronic pain conditions, whether plastic change occurs or not within the S1, and whether and how the plastic change contributes to chronic pain behavior, is unknown. Here, we provide the first evidence that intra-regional remodeling within the mouse S1 accelerates chronic pain behavior by modulating neuronal activity in the ACC, one of the important cortical areas for chronic pain. Using two-photon Ca(2+) imaging, we found that the spontaneous activity of layer 2/3 neurons in the S1 and then response to sensory and layer 4 stimulations increased under chronic pain conditions. In addition, pharmacological attenuation and facilitation of S1 activity attenuated and facilitated the chronic pain behavior, respectively. Furthermore, electrical response of the ACC to peripheral stimulation successfully correlated with S1 neuronal activity, and inhibition of ACC activity alleviated the mechanical allodynia. The present results will provide development of efficient therapeutic strategies against chronic pain by focusing on the S1 and ACC.


Differential Diagnosis and Precision Therapy of Two Typical Malignant Cutaneous Tumors Leveraging Their Tumor Microenvironment: A Photomedicine Strategy.
Elevated hydrogen peroxide (H2O2) in biological tissues is generally recognized to be relevant to the carcinogenesis process that regulates the proliferative activity of cancer cells and the transformation of malignant features. Inspired by this observation, it can be hypothesized that imaging H2O2 in the tumor microenvironment (TME) could help diagnose tumor types and malignancy, and even guide precise therapy. Thus, in this study, a noninvasive photomedicine strategy is demonstrated that leverages the different levels of H2O2 in the TME, and two representative skin cancers, malignant melanoma (MM, clinically higher incidence of metastasis and recurrence) and cutaneous squamous cell carcinoma (cSCC, relatively less dangerous), are differentially diagnosed. The working probe used here is one we previously developed, namely, intelligent H2O2 responsive ABTS-loaded HRP@Gd nanoprobes (iHRANPs). In this study, iHRANPs have advantages over ratiometric imaging due to their bimodal imaging elements, in which the inherent magnetic resonance imaging (MR) mode can be used as the internal imaging reference and the H2O2 responsive photoacoustic (PA) imaging modality can be used for differential diagnosis. Results showed that after intravenous injection of iHRANPs, the tumor signals on both MM and cSCC are obviously enhanced without significant difference under the MR modality. However, under the PA modality, MM and cSCC can be easily distinguished with obvious variations in signal enhancement. Particularly, guided by PA imaging, photothermal therapy (PTT) can be precisely applied on MM, and a strong antitumor effect was achieved owing to the excessive H2O2 in the TME of MM. Furthermore, exogenous H2O2 was injected into cSCC to remedy H2O2 deficiency in the TME of cSCC, and an evident therapeutic efficacy on cSCC can also be realized. This study demonstrated that MM can be differentially diagnosed from cSCC by noninvasive imaging of H2O2 in the TME with iHRANPs; meanwhile, it further enabled imaging-guided precision PTT ablation, even for those unsatisfactory tumor types (cSCC) through exogenously delivering H2O2.


Composition of gut microbiota and its correlations with neurological, intestinal, cardiovascular and metabolic diseases.
The intestinal microbiota is a microenvironment that has been the subject of studies for several decades. Over time, it has been reconsidered as a possible cofactor of multiple acute and chronic human diseases. In fact, alterations of the intestinal bacterial flora have been found in various neurological diseases. There are three modes of interaction between the intestinal microbiota and the gut-brain-axis: chemical signals, neural pathways and immune system. Even at the gastrointestinal level, the gut microbiota plays certainly an important role in the etiopathogenesis of chronic intestinal inflammatory diseases but also in irritable bowel syndrome. An important correlation has also been demonstrated with non-alcoholic fatty liver disease, as well as in other metabolic, cardiovascular and oncological diseases. Bacteria, viruses, fungi and various microorganisms that normally reside in our intestines can also be called into question as protective factors against these diseases. All this evidence leads researchers to consider the gut microbiota as a key element in the determination of aforementioned diseases. Therefore, it would be foreseeable in the future to associate the use of probiotics with the therapies used in the treatment of all these diseases. In this review we have condensed the main current knowledge regarding the link between the most frequent diseases and the gut microbiota.


Effect of influenza-induced fever on human bioimpedance values.
Bioelectrical impedance analysis (BIA) is a widely used technique to assess body composition and nutritional status. While bioelectrical values are affected by diverse variables, there has been little research on validation of BIA in acute illness, especially to understand prognostic significance. Here we report the use of BIA in acute febrile states induced by influenza. Bioimpedance studies were conducted during an H1N1 influenza A outbreak in Venezuelan Amerindian villages from the Amazonas. Measurements were performed on 52 subjects between 1 and 40 years of age, and 7 children were re-examined after starting Oseltamivir treatment. Bioelectrical Impedance Vector Analysis (BIVA) and permutation tests were applied. For the entire sample, febrile individuals showed a tendency toward greater reactance (p=0.058) and phase angle (p=0.037) than afebrile individuals, while resistance and impedance were similar in the two groups. Individuals with repeated measurements showed significant differences in bioimpedance values associated with fever, including increased reactance (p<0.001) and phase angle (p=0.007), and decreased resistance (p=0.007) and impedance (p<0.001). There are bioelectrical variations induced by influenza that can be related to dehydration, with lower extracellular to intracellular water ratio in febrile individuals, or a direct thermal effect. Caution is recommended when interpreting bioimpedance results in febrile states.


[Curcumin reduces paraquat-induced oxidative injury in A549 cells by activation of the Nrf2-ARE pathway].
To investigate the protective effect of curcumin (CU) on type II alveolar epithelial cells (A549 cells) during paraquat (PQ)-induced oxidative damage and its underlying mechanism. Routinely cultured A549 cells were divided into blank control group, CU control group, PQ group, and PQ+Cu group to receive respective treatments for 24 h. Cell viability was determined by MTT assay. The NFE2L2 expression in A549 cells was measured by RT-PCR and Western blot. The activities of the heme oxygenase-1 (HO-1) and NAD (P) H: quinone oxidoreductase 1 (NQO-1) in cells and the superoxide dismutase (SOD) and catalase (CAT) in supernatant, as well as malondialdehyde (MDA) content, were measured by enzyme-linked immunosorbent assay. After siRNA depletion of Nrf2, the protective effect of CU on A549 cells during PQ-induced oxidative damage was evaluated. PQ, even at a dose of 0.1 mmol/L, could significantly suppress the viability of A549 cells in a dose-dependent manner. CU showed no significant inhibitory effect on the viability of A549 cells when given at a dose below 160 ümol/L. Compared with the blank control group, the PQ group had significantly decreased SOD activity and significantly increased CAT activity and MDA content after 24-h exposure to 800 ümol/L PQ (P < 0.05 or P < 0.01). Thanks to pretreatment with 80 ümol/L CU, the PQ+CU group had significantly increased SOD and CAT activities and significantly decreased MDA content compared with the PQ group (P < 0.01). Compared with the blank control group, the PQ group had significantly increased expression of NFE2L2 and its downstream factors HO-1 and NQO-1 (P < 0.01), while the PQ+CU group had significantly higher expression of NFE2L2, HO-1,and NQO-1 than the PQ group (P < 0.01).Compared with the PQ+CU group, the CU+PQ+NFE2L2siRNA group had significantly decreased SOD and CAT activities and significantly increased MDA content (P < 0.01). Low-dose CU significantly reduces the PQ-induced oxidative damage in A549 cells in vitro by activation of the Nrf2-ARE pathway.


Transcription, beta-like DNA polymerases and hypermutation.
This paper discusses two aspects of immunoglobulin (Ig) gene hypermutation. In the first approach, a transcription termination signal is introduced in an Ig light chain transgene acting as a mutation substrate, and transgenic lines are generated with control and mutant transgenes integrated in tandem. Analysis of transcription levels and mutation frequencies between mutant and control transgenes clearly dissociates transcription elongation and mutation, and therefore argues against models whereby specific pausing of the RNA polymerase during V gene transcription would trigger an error-prone repair process. The second part reports the identification of two novel beta-like DNA polymerases named Pol lambda and Pol mu, one of which (Pol mu) represents a good candidate for the Ig mutase due to its higher lymphoid expression and its similarity with the lymphoid enzyme terminal deoxynucleotidyl transferase. Peculiar features of the expression of this gene, including an unusual splicing variability and a splicing inhibition in response to DNA-damaging agents, are discussed.


Red Cell Distribution Width to Platelet Ratio Is Associated with Increasing In-Hospital Mortality in Critically Ill Patients with Acute Kidney Injury.
Inflammation plays a key role in the pathophysiology and progression of acute kidney injury (AKI). Red cell distribution width (RDW) to platelet ratio (RPR) is a novel inflammatory index, and its prognostic effect on critically ill patients with AKI is rarely investigated. This work is aimed at investigating the association between RPR and in-hospital mortality in these patients. Data were extracted from the Medical Information Mart for Intensive Care III database. All-cause death during hospitalization was selected as the primary outcome. Receiver operating characteristic (ROC) curve was used to determine the optimal cut-off value, and the area under the curve (AUC) was applied to compare predictive ability among different indices. Cox proportional hazard models were utilized to assess the association between RPR and in-hospital mortality. Restricted cubic spline analysis for multivariate Cox model was performed to explore the shape of the relationship between RPR and mortality. A total of 24,166 critically ill patients with AKI were included. The relationship of RPR and in-hospital mortality was nonlinear with a trend to rise rapidly and then gradually. For mortality prediction, RPR had the optimal cut-off value of 0.093, of which the AUC was 0.791 (95% confidence interval (CI): 0.773-0.810), which was higher than those of RDW, platelet, sequential organ failure assessment score, simplified acute physiology score II, neutrophil to lymphocyte ratio, and platelet to lymphocytes ratio. After adjustments for various confounders, high RPR showed a significant association with increased mortality with hazard ratios of 1.46 (95% CI: 1.40-1.55) for categorical variable and 1.88 (95% CI: 1.80-1.97) for continuous variables in the fully adjusted model. Elevated RPR on admission is substantially associated with high risk of in-hospital mortality in critically ill patients with AKI and thus may serve as a novel predictor of prognosis for these patients.


Acorn Barnacles Secrete Phase-Separating Fluid to Clear Surfaces Ahead of Cement Deposition.
Marine macrofoulers (e.g., barnacles, tubeworms, mussels) create underwater adhesives capable of attaching themselves to almost any material. The difficulty in removing these organisms frustrates maritime and oceanographic communities, and fascinates biomedical and industrial communities seeking synthetic adhesives that cure and hold steadfast in aqueous environments. Protein analysis can reveal the chemical composition of natural adhesives; however, developing synthetic analogs that mimic their performance remains a challenge due to an incomplete understanding of adhesion processes. Here, it is shown that acorn barnacles (Amphibalanus (=Balanus) amphitrite) secrete a phase-separating fluid ahead of growth and cement deposition. This mixture consists of a phenolic laden gelatinous phase that presents a phase rich in lipids and reactive oxygen species at the seawater interface. Nearby biofilms rapidly oxidize and lift off the surface as the secretion advances. While phenolic chemistries are ubiquitous to arthropod adhesives and cuticles, the findings demonstrate that A. amphitrite uses these chemistries in a complex surface-cleaning fluid, at a substantially higher relative abundance than in its adhesive. The discovery of this critical step in underwater adhesion represents a missing link between natural and synthetic adhesives, and provides new directions for the development of environmentally friendly biofouling solutions.


Numerical study of the effect of thiol-disulfide exchange in the cluster phase of beta-lactoglobulin aggregation.
We report a numerical study of beta-lactoglobulin aggregation using grand canonical Monte Carlo simulations of a simple lattice model in which the proteins are represented by a single lattice point and interact via a sum of a short-ranged attraction, and a long-ranged screened electrostatic repulsion. For certain values of the potential parameters we observe the so-called cluster phase, in which protein aggregates of finite size repel each other. The properties of the cluster phase are dependent on the salt concentration, the charge of the protein, and the strength of the short-ranged attraction. Disulfide bridges are modeled by covalent bonds between the lattice points, and can exchange with free thiols. Allowing the thiol-disulfide exchange leads to a severe lowering in the chemical potential of the cluster transition, or equivalently, a lower monomer density. Moreover, we find that the disulfide bridges (or the free thiol groups) are not uniformly distributed over the aggregate. The free thiol groups are significantly more abundant on the surface than in the core of the aggregate, making the surface more reactive than the inner core. This finding might explain why films made of beta-lactoglobulin by cold gelation, after resolvation, reconstitute finite aggregates rather than a monomer solution.


The first 6 months: developing a user-informed anticipatory counselling video about the levonorgestrel intrauterine system.
The levonorgestrel intrauterine system (LNG-IUS) is a contraceptive method that can cause irregular bleeding and cramping during the first 6 months of use. Expected side effects are common reasons given for LNG-IUS device discontinuation within 12 months of use. Anticipatory counselling regarding expected LNG-IUS side effects may reduce method discontinuation and improve patient satisfaction. Educational videos could improve anticipatory counselling for LNG-IUS users; however, none have been developed. This article describes the process of developing an anticipatory counselling video with input from women in the first 6 months of LNG-IUS use and from family planning (FP) experts. We used a participatory, iterative process to develop an anticipatory counselling video about the first 6 months of LNG-IUS use. We developed a preliminary draft using evidence from the published literature. We sought feedback from 11 FP experts and 49 LNG-IUS users to revise the script before creating the final video. FP experts suggested balancing negative with positive information and using colloquial language. LNG-IUS users requested more detailed information on the LNG-IUS mechanism of action and expected side effects in the form of statistics, infographic animations, testimonials from LNG-IUS users, and technical as opposed to colloquial language. The final video is 6 min in length and features seven LNG-IUS users, three physicians, and infographic animations. Including input from FP experts and LNG-IUS users in the development process resulted in a 6-min anticipatory counselling video that will be piloted among patients on the day of their LNG-IUS insertion.


Genetic Variation at the ADAMTS7 Locus is Associated With Reduced Severity of Coronary Artery Disease.
Genome-wide association studies identified ADAMTS7 as a risk locus for coronary artery disease (CAD). Functional studies suggest that ADAMTS7 may promote cellular processes in atherosclerosis. We sought to examine the association between genetic variation at ADAMTS7 and measures of atherosclerosis using histological, angiographic, and clinical outcomes data. The lead CAD-associated single-nucleotide polymorphism rs3825807 at the ADAMTS7 locus was genotyped. The G allele (reduced ADAMTS7 function) was associated with a smaller fibrous cap (P=0.017) and a smaller percentage area of α-actin (smooth muscle cell marker) in the intima (P=0.017), but was not associated with calcification or plaque thickness, following ex vivo immunohistochemistry analysis of human coronary plaques (n=50; mean age 72.2±11.3). In two independent cohorts (Southampton Atherosclerosis Study [n=1359; mean age 62.5±10.3; 70.1% men] and the Emory Cardiovascular Biobank [EmCAB; n=2684; mean age 63.8±11.3; 68.7% men]), the G allele was associated with 16% to 19% lower odds of obstructive CAD (Southampton Atherosclerosis Study: odds ratio, 0.81; 95% confidence interval, 0.67-0.98; EmCAB: odds ratio, 0.84; 95% confidence interval, 0.75-0.95) with similar effects for multivessel, left anterior descending, and proximal CAD. Furthermore, each copy of the G allele was associated with lower angiographic severity Gensini score (Southampton Atherosclerosis Study, P=0.026; EmCAB, P<0.001), lower Sullivan Extent score (Southampton Atherosclerosis Study, P=0.029; EmCAB, P<0.001), and a 23% lower risk of incident revascularization procedures (EmCAB: hazard ratio, 0.76; 95% confidence interval, 0.59-0.98). There were no associations with all-cause mortality or incident myocardial infarction. Genetic variation at the ADAMTS7 locus is associated with several complementary CAD phenotypes, supporting the emerging role of ADAMTS7 in atherosclerosis and may represent a potential drug target.


A Deeper Understanding of Metal Nucleation and Growth in Rechargeable Metal Batteries Through Theory and Experiment.
Lithium and sodium metal batteries continue to occupy the forefront of battery research. Their exceptionally high energy density and nominal voltages are highly attractive for cutting-edge energy storage applications. Anode-free metal batteries are also coming into the research spotlight offering improved safety and even higher energy densities than conventional metal batteries. However, uneven metal nucleation and growth which leads to dendrites continues to limit the commercialisation of conventional and anode-free metal batteries alike. This review connects models and theories from well-established fields in metallurgy and electrodeposition to both conventional and anode-free metal batteries. These highly applicable models and theories explain the driving forces of uneven metal growth and can inform future experiment design. Finally, the models and theories that are most relevant to each anode-related cell component are identified. Keeping these specific models and theories in mind will assist with rational design for these components.


Improving Tumor Specificity and Anticancer Activity of Dasatinib by Dual-Targeted Polymeric Micelles.
To improve tumor targetability and drug efficacy and decrease drug resistance of dasatinib (DSB), the multifunctional micellar nanoparticles that combined the matrix metalloproteinase 2 (MMP2)-sensitive tumor (site) targeting with folate receptor-mediated tumor (cell) targeting were developed. Two major functional polymers, polyethylene glycol (5000 Da)-MMP2-sensitive peptide-phosphoethanolamine (PEG5k-pp-PE) and folic acid-polyethylene glycol (2000 Da)-phosphoethanolamine (FA-PEG2k-PE), were synthesized to construct the dual-targeted micellar nanoparticles (MMP/FR micelles). In the absence of MMP2, the FA was shielded by PEG5k and the MMP/FR micelles showed low bioactivity. In the presence of MMP2, the nanoparticulate structure, stability, and cargo release profile of the MMP/FR micelles were not significantly affected, however, the MMP2-mediated PEG5k deshielding and FA exposure remarkably increased the cellular uptake and anticancer activity of the micelles in the MMP2 and FR expressing (MMP2+/FR+) cells, including multidrug resistant (MDR) cancer cells, rather than the MMP2- and FR- cells. In the 3D MDR tumor spheroids, the significant MMP2-dependent tissue penetration, uptake and cytotoxicity of the MMP/FR micelles were also observed. Furthermore, in the in vivo biodistribution study, the MMP2 and FR dual targeting strategy could significantly prolong the systemic circulation, decrease the nonspecific distribution in nontumor tissues, and increase the tumor accumulation of the polymeric micelles in a melanoma xenograft mouse model. The MMP2-sensitive FR-targeted micelles might have great potential as a tumor-targeted platform for delivery of molecular targeted therapeutics.


Astroglial Cx30 differentially impacts synaptic activity from hippocampal principal cells and interneurons.
Astrocytes play important roles in brain function via dynamic structural and functional interactions with neurons. Yet the underlying mechanisms remain poorly defined. A typical feature of astrocytes is the high expression of connexins, which mediate their extensive intercellular communication and regulate their structural properties. In particular, connexin 30 (Cx30), one of the two connexins abundantly expressed by astrocytes, was recently shown to be a critical regulator of excitatory synaptic transmission by controlling the astroglial coverage of synapses. However, the role of Cx30 in the regulation of inhibitory synaptic transmission and excitatory/inhibitory balance remains elusive. Here, we investigated the role of astroglial Cx30 on the electrophysiological and morphological properties of five classes of hippocampal CA1 stratum oriens and pyramidale neurons, defined by the unsupervised Ward's clustering. Using Cx30 knockout mice, we found that Cx30 alters specific properties of some subsets of CA1 interneurons, such as resting membrane potential and sag ratio, while other parameters, such as action potential threshold and saturation frequency, were more frequently altered among the different classes of neurons. The excitation-inhibition balance was also differentially and selectively modulated among the different neuron subtypes. Only slight morphological differences were observed on reconstructed neurons. Altogether, these data indicate that Cx30 differentially alters the electrophysiological and morphological properties of hippocampal cell populations, and modulates both their excitatory and inhibitory inputs. Astrocytes, via Cx30, are thus active modulators of both excitatory and inhibitory synapses in the hippocampus.


[Type 1 diabetic patients evolution to hypertension].
Check the evolution of type 1(T1) diabetic patients to hypertension and prehypertension and baseline factors related to final blood pressure levels (BP). Observational study involving 127 T1 diabetic patients submitted to clinical and laboratory evaluation and followed by for 5 (2.4-9.2) years. From the initially normotensive patients, 21.7% developed prehypertension, 4.7% developed hypertension and 73.6% remained with normal BP. From the prehypertensive patients, 35% returned to normal BP, 50% remained prehypertensive and 15% developed hypertension. The relative risk for hypertension development was 3.2 (0.8-12.3) in the prehypertensive compared to the normotensive group. The prevalence of prehypertension and hypertension increased from 15.7% to 26% and 0.8% to 7% respectively. Levels of serum creatinine predicted final levels of systolic and diastolic BP. It is emphasized the importance of renal function and BP evaluation even when they are in normal range to minimize the deleterious effects of hypertension in the development of nephropathy and cardiovascular disease.


Process of forgiveness in the recovery of Chinese women survivors of intimate partner violence: Empowerment, transformation, and integration.
Forgiveness has been found one substantial element in the recovery for women survivors from intimate partner violence following the termination of the abusive relationship. To further investigate the details of forgiveness in this specific context, the present study explored the process of forgiveness using grounded theory. In-depth and semi-structured interviews were conducted with 25 Chinese women survivors of IPV. The findings suggest that forgiveness is a strength-based process including empowerment, transformation, and integration phases. In the empowerment phase, survivors obtain strength at the intrapersonal, behavioural, and interpersonal levels. In the transformation phase, survivors complete cognitive transformation for their IPV experiences and emotional transformation towards former partners. In the integration phase, survivors-now freed from the past-reflect upon and apply the changes they have undergone. Two trajectories in the process were found. One trajectory is going through stages sequentially and the other trajectory is experiencing back and forth between empowerment and transformation stages before moving into the integration stage. The study's findings broaden our knowledge of the strength-based forgiveness process that women survivors of IPV undergo during recovery. Practitioners and policymakers could develop programmes and policies that support forgiveness by holistically facilitating their recovery and empowerment like assistance in dealing with life difficulties and promoting their reconnection with social networks. To improve the transferability and validity of the findings, the forgiveness of survivors of IPV could be explored in a diverse sample (e.g., survivors with low educational background or live in the rural area).


Effects of cyclofenil diphenol, an agent which disrupts Golgi structure, on proteoglycan synthesis in chondrocytes.
1. Cyclofenil diphenol (F6060), a weak non-steroidal oestrogen, was shown previously to inhibit [35S]proteoglycan synthesis [Mason, Lineham, Phillipson & Black (1984) Biochem. J. 223, 401-412] and to induce fragmentation of the Golgi apparatus into small vesicles [Lancaster, Fryer, Griffiths & Mason (1989) J. Cell Sci. 92, 271-280] in cultures of Swarm chondrosarcoma chondrocytes. Two structurally related compounds, F6204 and F6091, show a similar concentration-related effect, with complete inhibition of [35S]proteoglycan synthesis at 90 micrograms/ml. The apparent [3H]protein synthesis is only approx. 40% inhibited with [3H]lysine as precursor. Stilboestrol, clomiphene and tamoxiphen are also potent inhibitors of [35S]proteoglycan synthesis. 2. Syntheses of chondroitin 4-[35S]sulphate and chondroitin 6-[35S]sulphate, which are Golgi-mediated events, are inhibited 40-68% and 3-48% respectively by concentrations of cyclofenil between 50 and 70 micrograms/ml. [3H]Hyaluronan synthesis, which occurs by a different mechanism at the plasma membrane, is inhibited by 47-66%. These results suggest that cyclofenil may act via more than one inhibitory mechanism. Cyclofenil diphenol inhibits polymerization of chondroitin sulphate on to p-nitrophenyl beta-xyloside even when the chondrocytes are loaded with the initiator prior to treatment. 3. Cyclofenil diphenol interferes with the cellular uptake of amino acids via the system A carrier, as shown by inhibition of uptake of methylaminoisobutyric acid, a specific substrate for this system. The drug had no effect on the uptake of 2-deoxyglucose by the cells. 4. Cyclofenil diphenol (90 micrograms/ml) caused a decrease in the pool size of UDP-N-acetylglucosamine, UDP-N-acetylgalactosamine and UDP-hexoses, but this was insufficient to account for the accompanying profound inhibition of [35S]proteoglycan synthesis. Entry of [3H]glucosamine into the cell and into the UDP-N-acetylhexosamine pool did not appear to be affected. 5. Cyclofenil diphenol inhibited the substitution of 3H-labelled proteoglycan core protein with chondroitin sulphate chains. Core protein was identified in treated cultures on the basis of immunoprecipitation with an antiserum against the hyaluronate-binding region and distinguished from precipitated proteoglycan on SDS/PAGE.


Achieving consensus for clinical trials: the REiNS International Collaboration.
The neurofibromatoses (NF)--including neurofibromatosis 1 (NF1), neurofibromatosis 2 (NF2), and schwannomatosis--are related tumor-suppressor syndromes characterized by a predisposition to multiple tumor types and other disease manifestations, which often result in functional disability, reduced quality of life, pain, and, in some cases, malignancy. With increasing knowledge of the biology and pathogenesis of NF, clinical trials with targeted agents directed at NF tumors have become available. Most clinical trials for patients with NF have used designs and endpoints similar to oncology trials. However, differences in the disease manifestations and natural history of NF (compared to cancers) require the development of new designs and endpoints to perform meaningful NF clinical trials. The Response Evaluation in Neurofibromatosis and Schwannomatosis (REiNS) International Collaboration was established in 2011 at the Children's Tumor Foundation meeting to achieve consensus within the NF community about the design of future clinical trials, with a specific emphasis on endpoints. The REiNS Collaboration includes 7 working groups that focus on imaging of tumor response; functional, visual, patient-reported, and neurocognitive outcomes; whole-body MRI; and disease biomarkers. This supplement includes the first series of recommendations by the REiNS Collaboration. The hope is that these recommendations will be used by members of the group and by researchers outside of the REiNS International Collaboration to standardize the measurement of outcomes and thus improve clinical trials for patients with NF. Ultimately, we plan to engage industry partners and national regulatory agencies in this process to facilitate the approval of drugs for patients with NF.


Age affects the contraction-induced mitochondrial redox response in skeletal muscle.
Compromised mitochondrial respiratory function is associated with advancing age. Damage due to an increase in reactive oxygen species (ROS) with age is thought to contribute to the mitochondrial deficits. The coenzyme nicotinamide adenine dinucleotide in its reduced (NADH) and oxidized (NAD(+)) forms plays an essential role in the cyclic sequence of reactions that result in the regeneration of ATP by oxidative phosphorylation in mitochondria. Monitoring mitochondrial NADH/NAD(+) redox status during recovery from an episode of high energy demand thus allows assessment of mitochondrial function. NADH fluoresces when excited with ultraviolet light in the UV-A band and NAD(+) does not, allowing NADH/NAD(+) to be monitored in real time using fluorescence microscopy. Our goal was to assess mitochondrial function by monitoring the NADH fluorescence response following a brief period of high energy demand in muscle from adult and old wild-type mice. This was accomplished by isolating whole lumbrical muscles from the hind paws of 7- and 28-month-old mice and making simultaneous measurements of force and NADH fluorescence responses during and after a 5 s maximum isometric contraction. All muscles exhibited fluorescence oscillations that were qualitatively similar and consisted of a brief transient increase followed by a longer transient period of reduced fluorescence and, finally, an increase that included an overshoot before recovering to resting level. Compared with the adult mice, muscles from the 28 mo mice exhibited a delayed peak during the first fluorescence transient and an attenuated recovery following the second transient. These findings indicate an impaired mitochondrial capacity to maintain NADH/NAD(+) redox homeostasis during contractile activity in skeletal muscles of old mice.


Efficacy of nonestrogenic hot flash therapies among women stratified by breast cancer history and tamoxifen use: a pooled analysis.
Various nonestrogenic therapies have been found to be effective in mitigating hot flashes, but it has been unclear whether the efficacy varies by whether women have had breast cancer and/or were taking tamoxifen. This study used data from Mayo Clinic/North Central Cancer Treatment Group clinical trials that evaluated the efficacy of any nonestrogenic agent for hot flashes and had information on breast cancer history or tamoxifen use. Statistically significant changes from the fourth treatment week versus the baseline week, using individual patient data, were assessed using Student's t test. A total of 1,396 women from 20 hot flash studies were eligible for analysis. Overall, women without breast cancer had a similar percentage of baseline hot flash score at week 4, as did those with breast cancer (53% vs 50%, P = 0.92). Women who were not taking tamoxifen had a significantly lower percentage of hot flash score at week 4 as compared with those who used tamoxifen (54% vs 61%, P = 0.01). However, this was due to a higher reduction in hot flash scores in the placebo arms among women not receiving tamoxifen; the percentage reduction in hot flash scores at week 4 from baseline in the active therapy arms of the randomized placebo-controlled trials (ie, excluding placebo arms) was similar among the tamoxifen users and nonusers (difference in mean percentage reduction, 5.7; 95% CI, -1.76 to 13.16). Some nonestrogenic therapies seem to be useful for reducing hot flashes, irrespective of the etiology of hot flashes.


Quantification of progressive joint space narrowing in osteoarthritis of the hip: longitudinal analysis of the contralateral hip after total hip arthroplasty.
The rate of progressive joint space narrowing in the contralateral hip after total hip arthroplasty (THA) for osteoarthritis (OA) and the factors which may predispose patients to more aggressive joint space narrowing remain undefined. The current study sought to evaluate the rate and pattern of, and risk factors for, progressive joint space narrowing in the contralateral hip after THA for OA. Each patient who underwent THA for OA in 1984-1985 was followed up longitudinally, and annual anteroposterior (AP) pelvis radiographs were obtained. The radiographic joint space width (JSW) of each contralateral hip joint was quantified, and the rates of JSW narrowing were determined. Evaluation of potential risk factors for accelerated progression of joint space narrowing included age, sex, side of surgery, weight, height, body mass index (BMI), hip pain, etiology of OA, and Kellgren/Lawrence radiographic grade. Ninety-nine patients and 619 AP pelvis radiographs were evaluated. The median initial JSW was 3.48 mm (interquartile range 1.55). JSW declined in a linear manner at a median rate of 0.10 mm/year. The rate of decline between baseline and followup in 20 months was predictive of the overall slope. Two subpopulations were identified. Eighty-five percent of patients maintained a slow decline in JSW (< or =0.2 mm/year), and 15% exhibited an accelerated decline in JSW (>0.2 mm/year). Kellgren/ Lawrence radiographic grade > or =2 and a diagnosis of primary OA were each associated with a more rapid decline in JSW (P = 0.006 and P = 0.02, respectively). Initial JSW, age, sex, weight, height, BMI, and hip pain were not risk factors for rapid decline in JSW. Radiographic hip JSW may be reliably quantified and followed up longitudinally using standard AP radiographs. Progression of JSW narrowing in the contralateral hip after THA for OA proceeds in a linear manner over several years. A subpopulation of patients with accelerated narrowing of contralateral JSW may be identified within 20 months, and may represent a suitable population with which to assess the potential efficacy of new disease-modifying agents.


Oxymatrine Inhibits Renal Cell Carcinoma Progression by Suppressing β-Catenin Expression.
Oxymatrine (OMT) has been identified to possess immunomodulatory, antiinflammatory and anticancer properties. This study aimed to investigate its precise function and the underlying molecular mechanisms in renal cell carcinoma progression. The antineoplastic effect of oxymatrine was investigated by CCK-8 assay, cell cycle analysis, apoptosis assay, wound healing experiment, transwell assay, and drug-sensitivity analysis in renal cancer cells following oxymatrine treatment. The modulation of oxymatrine on β-catenin was analyzed through western blot and immunofluorescence assay. β-catenin overexpression was employed to determine the key role of β-catenin in oxymatrine-inhibited renal cell carcinoma in vitro. In addition, animal model was established to investigate the effect of oxymatrine on tumor growth in vivo. Oxymatrine inhibited renal cell carcinoma progression in vitro, including cell proliferation, apoptosis, migration, invasion and chemotherapy sensitivity. Further mechanistic studies demonstrated that oxymatrine exerted its antineoplastic effect through suppressing the expression of β-catenin. Moreover, in nude mice model, oxymatrine exhibited remarkable inhibition of tumor growth, which was consistent with our in vitro results. Our findings illuminate oxymatrine as an effective antitumor agent in renal cell carcinoma, and suggest it a promising therapeutic application in renal cell carcinoma treatment.


Interpreting and understanding meta-analysis graphs--a practical guide.
Ideally, clinical decision making ought to be based on the latest evidence available. However, to keep abreast with the continuously increasing number of publications in health research, a primary health care professional would need to read an insurmountable number of articles every day covered in more than 13 million references and over 4800 biomedical and health journals in Medline alone. With the view to address this challenge, the systematic review method was developed. This article provides a practical guide for appraising systematic reviews for relevance to clinical practice and interpreting meta-analysis graphs as part of quantitative systematic reviews.


Environmental factors and psoriasis.
Psoriasis is a chronic relapsing disease characterized by variable clinical features. Several factors may exacerbate its manifestations, or even trigger the disease, such as traumatic injury to the skin, physical and psychological stress, cold weather, excessive alcohol intake, and drugs such as lithium and Beta-blockers. We describe the most common features of psoriasis and the exogenous factors that may induce, trigger or exacerbate the disease.


Functional analysis and regulation of the divergent spuABCDEFGH-spuI operons for polyamine uptake and utilization in Pseudomonas aeruginosa PAO1.
A multiple-gene locus for polyamine uptake and utilization was discovered in Pseudomonas aeruginosa PAO1. This locus contained nine genes designated spuABCDEFGHI (spu for spermidine and putrescine utilization). The physiological functions of the spu genes in utilization of two polyamines (putrescine and spermidine) were analyzed by using Tn5 transposon-mediated spu knockout mutants. Growth and uptake experiments support that the spuDEFGH genes specify components of a major ABC-type transport system for spermidine uptake, and enzymatic measurements indicated that spuC encodes putrescine aminotransferase with pyruvate as the amino group receptor. Although spuA and spuB mutants showed an apparent defect in spermidine utilization, the biochemical functions of the gene products have yet to be elucidated. Assays of lacZ fusions demonstrated the presence of agmatine-, putrescine-, and spermidine-inducible promoters for the spuABCDEFGH operon and the divergently transcribed spuI gene of unknown function. Since the observed induction effect of agmatine was abolished in an aguA mutant where conversion of agmatine into putrescine was blocked, putrescine or spermidine, but not agmatine, serves as the inducer molecule of the spuA-spuI divergent promoters. S1 nuclease mappings confirmed further the induction effects of the polyamines on transcription of the divergent promoters and localized the transcription initiation sites. Gel retardation assays with extracts from the cells grown on putrescine or spermidine demonstrated the presence of a polyamine-responsive regulatory protein interacting with the divergent promoter region. Finally, the absence of the putrescine-inducible spuA expression and putrescine aminotransferase (spuC) formation in the cbrB mutant indicated that the spu operons are regulated by the global CbrAB two-component system perhaps via the putative polyamine-responsive transcriptional activator.


Linear ubiquitination signals in adaptive immune responses.
Ubiquitin can form eight different linkage types of chains using the intrinsic Met 1 residue or one of the seven intrinsic Lys residues. Each linkage type of ubiquitin chain has a distinct three-dimensional topology, functioning as a tag to attract specific signaling molecules, which are so-called ubiquitin readers, and regulates various biological functions. Ubiquitin chains linked via Met 1 in a head-to-tail manner are called linear ubiquitin chains. Linear ubiquitination plays an important role in the regulation of cellular signaling, including the best-characterized tumor necrosis factor (TNF)-induced canonical nuclear factor-κB (NF-κB) pathway. Linear ubiquitin chains are specifically generated by an E3 ligase complex called the linear ubiquitin chain assembly complex (LUBAC) and hydrolyzed by a deubiquitinase (DUB) called ovarian tumor (OTU) DUB with linear linkage specificity (OTULIN). LUBAC linearly ubiquitinates critical molecules in the TNF pathway, such as NEMO and RIPK1. The linear ubiquitin chains are then recognized by the ubiquitin readers, including NEMO, which control the TNF pathway. Accumulating evidence indicates an importance of the LUBAC complex in the regulation of apoptosis, development, and inflammation in mice. In this article, I focus on the role of linear ubiquitin chains in adaptive immune responses with an emphasis on the TNF-induced signaling pathways.


Executing a Series of Zinc(II) Complexes of Homologous Schiff Base Ligands for a Comparative Analysis on Hydrolytic, Antioxidant, and Antibacterial Activities.
Six zinc(II) complexes, namely, [Zn(HL1H)Cl2] (1), [Zn(HL1H)Br2] (2), [Zn2(HL1H)2(OH)I2]·I (3), [Zn(HL2)Cl] (4), [Zn2(HL2)Br3] (5), and [Zn(HL2)I] (6) have been manufactured by using two homologous Schiff base ligands H2L1 and H2L2 for the purpose of perlustrating their phosphatase-like activity, antioxidant activity, and antibacterial activity. Complexes 1, 2, 4, and 5 have been reported earlier by us, whereas complexes 3 and 6 have been synthesized and structurally characterized by regular physicochemical methods The hydrolytic property of the six complexes has been evaluated by checking the hydrolysis of the P-O bond of a widely used substrate, namely, disodium salt of (para-nitrophenyl)phosphate (PNPP) in 97.5% (v/v) mixture of N,N-dimethylformamide and water (DMF-water). Complexes 2-5 have profound efficiency toward hydrolysis of phosphate ester bonds, and complexes 1 and 6 were noted to be inactive toward hydrolysis. Complex 3 displayed the highest efficacy among the six complexes. Additionally, antioxidant and antibacterial activities of the complexes were studied thoroughly. A detailed study of their antioxidant property revealed that complex 3 manifested superior radical scavenging activity, thus exhibiting the highest antioxidant property. The antibacterial activity was tested using four investigating bacteria, specifically Listeria monocytogenes ATCC19111, Staphylococcus aureus ATCC 700699, Salmonella typhimurium ATCC 23564, and Escherichia coli ATCC 25922 by determining minimum inhibitory concentration (MIC) values using the microdilution method. Here as well, complex 3 exhibited the highest activity to both Gram positive and Gram negative bacteria. The chemistry behind these experimental findings has been manifested by shedding light upon the structural features of the complexes. The suitable choice of ligand H2L1 where one methylene group is less than its homologous ligand and metal precursor (ZnI2) imparts a unique hydroxo-bridged molecular geometry and 2D hydrogen bonding network which in turn probably enhances the hydrolytic and biological activities of complex 3.


Simultaneous intrinsic signal imaging of auditory and visual cortex reveals profound effects of acute hearing loss on visual processing.
It has been suggested that primary sensory cortices do not work in isolation but receive subthreshold inputs originating from other senses. However, repercussions of an acute loss of one sense on multimodal sensory processing remain elusive. Here we investigated the early effects of acute hearing loss on visual processing in adult mice. For this, we developed a method to simultaneously map the primary auditory (A1) and visual cortex (V1) using periodic intrinsic optical imaging. We found that reducing sound evoked A1 responsiveness due to the induction of conductive hearing loss (CHL) led to a concomitant increase of visually driven V1 activity. Accordingly, using the neuronal activity marker c-fos we found the number of stained pyramidal cells to be increased in V1 layer 2/3 after CHL. In contrast, numbers of c-fos positive parvalbumin (PV) and somatostatin (SOM) expressing inhibitory neurons were reduced after CHL. Finally, we adapted the periodic intrinsic imaging method to determine V1 contrast and spatial frequency tuning. Using this method we could show that visual acuity and contrast sensitivity were improved after CHL. In addition, retrograde tracing experiments revealed direct anatomical projections from A1 to V1 which could potentially serve as a substrate for the observed effects. In summary, our results suggest that CHL rapidly disrupts the functional interplay between A1 and V1 leading to altered visually evoked V1 responses.


Overexpression of microRNA-206 in the skeletal muscle from myotonic dystrophy type 1 patients.
MicroRNAs are highly conserved, noncoding RNAs involved in post-transcriptional gene silencing. They have been shown to participate in a wide range of biological processes, including myogenesis and muscle regeneration. The goal of this study is to test the hypothesis that myo-miRs (myo = muscle + miR = miRNA) expression is altered in muscle from patients affected by myotonic dystrophy type 1 (DM1), the most frequently inherited neuromuscular disease in adults. In order to gain better insights about the role of miRNAs in the DM1 pathogenesis, we have also analyzed the muscular expression of miR-103 and miR-107, which have been identified in silico as attractive candidates for binding to the DMPK mRNA. To this aim, we have profiled the expression of miR-133 (miR-133a, miR-133b), miR-1, miR-181 (miR-181a, miR-181b, miR-181c) and miR-206, that are specifically induced during myogenesis in cardiac and skeletal muscle tissues. miR-103 and miR-107, highly expressed in brain, heart and muscle have also been included in this study. QRT-PCR experiments have been performed on RNA from vastus lateralis biopsies of DM1 patients (n = 7) and control subjects (n = 4). Results of miRNAs expression have been confirmed by Northern blot, whereas in situ hybridization technique have been performed to localize misexpressed miRNAs on muscle sections from DM1 and control individuals. Only miR-206 showed an over-expression in 5 of 7 DM1 patients (threshold = 2, fold change between 1.20 and 13.22, average = 5.37) compared to the control group. This result has been further confirmed by Northern blot analysis (3.37-fold overexpression, R2 = 0.89). In situ hybridization localized miR-206 to nuclear site both in normal and DM1 tissues. Cellular distribution in DM1 tissues includes also the nuclear regions of centralized nuclei, with a strong signal corresponding to nuclear clumps. This work provides, for the first time, evidences about miRNAs misexpression in DM1 muscle tissues, adding a new element in the pathogenesis of this complex genetic disease.


Magnetically-responsive electrophoretic silica organosols.
Silica nanoparticles can be dispersed in organic solvents (organosols) using surfactants, such as didodecyldimethylammonium bromide (DDAB). DDAB analogs prepared with lathanide tetrahalide counterions, either a high-magnetic moment ion (HoCl3Br, DDAH) or low-magnetic moment one (NdCl3Br, DDAN), are expected to produce charged particles, but only DDAH-stabilized dispersions are expected to be magnetically responsive. Phase-analysis light scattering (PALS) measurements have been performed to determine the charge on DDAH- and DDAN-stabilized organosols. Magnetic sedimentation experiments have been performed to determine whether or not the silica dispersions are magnetically responsive. Sedimentation was monitored both by visual observations and UV-vis spectroscopy. Both DDAH and DDAN organosols are negatively charged, the same as DDAB-stabilized nanoparticles. The DDAH-stabilized nanoparticles are found to respond to a magnetic field, whereas the DDAN-stabilized nanoparticles do not. This opens up possibilities for creating nanoparticle dispersions in nonpolar solvents which can be tailored to respond to desired external stimuli.