Family therapy techniques and one-year clinical outcomes among adolescents in usual care for behavior problems.
This study tested whether use of family therapy (FT) techniques predicted long-term clinical outcomes in usual care for adolescents enrolled in either family-based or non-family-based treatment for conduct and substance use problems. Participants included 70 adolescents (53% female; mean age 15.4 years) from diverse backgrounds (64% Hispanic, 16% African American, 11% multiracial) assessed at baseline and 3-, 6-, and 12-month follow-up. Utilization of FT techniques was assessed for 802 therapy sessions via a therapist-report tool that collected postsession data on delivery of core treatment techniques of the FT approach. Latent growth curve modeling was used to examine individual change over 12-month follow-up. FT technique scores, averaged across treatment sessions, were included as a predictor in latent growth models, controlling for adolescent age, sex, and ethnicity. More extensive use of core FT techniques predicted significant decreases in adolescent-reported delinquency and externalizing behavior and marginal decreases in substance use and parent-reported externalizing for cases in both family and nonfamily treatment. Greater use of FT techniques in routine care was associated with better long-term adolescent outcomes, regardless of whether clients attended services featuring family therapy or an alternative treatment approach. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


The myositis clinical phenotype associated with anti-Zo autoantibodies: a case series of nine UK patients.
It has been over 10 years since the first report of autoantibodies directed against phenylalanyl tRNA synthetase (anti-Zo) in a patient with features of the anti-synthetase syndrome. In that time no further cases have been published. Here we aim to characterize more fully the clinical phenotype of anti-Zo-associated myositis by describing the clinical features of nine patients. Anti-Zo was identified by protein-immunoprecipitation in patients referred for extended spectrum myositis autoantibody testing at our laboratory. Results were confirmed by immunodepletion using a reference serum. Medical records were retrospectively reviewed to provide detailed information of the associated clinical phenotype for all identified patients. Where possible, HLA genotype was imputed using Illumina protocols. Nine patients with anti-Zo were identified. The median age at disease onset was 51 years, and six patients were female. Seven patients had evidence of inflammatory muscle disease, seven of interstitial lung disease and six of arthritis. The reported pattern of interstitial lung disease varied with usual interstitial pneumonia, non-specific interstitial pneumonia and organizing pneumonia all described. Other features of the anti-synthetase syndrome such as RP and mechanics hands were common. HLA data was available for three patients, all of whom had at least one copy of the HLA 8.1 ancestral haplotype. Patients with anti-Zo presenting with features of the anti-synthetase syndrome and interstitial lung disease is a common finding. Like other myositis autoantibodies, there is likely to be a genetic association with the HLA 8.1 ancestral haplotype.


Association between specific periodontal pathogens, Toll-like receptor-4, and nuclear factor-κB expression in placental tissues of pre-eclamptic women with periodontitis.
The aim of the present study was to determine the association between the presence of specific periodontal pathogens, Toll-like receptor-4 (TLR-4), and nuclear factor-κB (NF-κB) expression in the placental tissues of pre-eclamptic women. Antenatal periodontal screening was performed in 25 normotensive pregnant women and 25 pre-eclamptic women. Subgingival plaque and placental tissue samples were collected from both groups and screened for the presence of Porphyromonas gingivalis (P. gingivalis), Tannerella forsythia, Aggregatibacter actinomycetemcomitans, and Prevotella intermedia (P. intermedia) using real-time polymerase chain reaction. The placental samples were also analyzed to quantify TLR-4 and NF-κB expression. The subgingival plaque samples of pre-eclamptic women showed significantly higher frequencies of P. intermedia. In the placental samples, P. gingivalis, P. intermedia, and the expression of TLR-4 and NF-κB were found to be at significantly higher levels compared to normotensive pregnant women. Using linear regression analysis, the expression of TLR-4 was significantly influenced by the presence of P. gingivalis (coefficient=3.176, 95% confidence interval [CI]: 367-5.986) and P. intermedia (coefficient=2.886, 95% CI: 0.77-5.696), whereas NF-κB expression was influenced only by the presence of P. intermedia (coefficient=2.220, 95% CI: 0.051-4.388) in the placental tissues of pre-eclamptic women. An association exists between P. gingivalis and P. intermedia with increased TLR-4 and NF-κB expression in the placenta of pre-eclamptic women with periodontitis.


Substituent and solvent effects on the UV-vis absorption spectrum of the photoactive yellow protein chromophore.
Solvent effects on the UV-vis absorption spectra and molecular properties of four models of the photoactive yellow protein (PYP) chromophore have been studied with ASEP/MD, a sequential quantum mechanics/molecular mechanics method. The anionic trans-p-coumaric acid (pCA(-)), thioacid (pCTA(-)), methyl ester (pCMe(-)), and methyl thioester (pCTMe(-)) derivatives have been studied in gas phase and in water solution. We analyze the modifications introduced by the substitution of sulfur by oxygen atoms and hydrogen by methyl in the coumaryl tail. We have found some differences in the absorption spectra of oxy and thio derivatives that could shed light on the different photoisomerization paths followed by these compounds. In solution, the spectrum substantially changes with respect to that obtained in the gas phase. The n → π1* state is destabilized by a polar solvent like water, and it becomes the third excited state in solution displaying an important blue shift. Now, the π → π1* and π → π2* states mix, and we find contributions from both transitions in S1 and S2. The presence of the sulfur atom modulates the solvent effect and the first two excited states become practically degenerate for pCA(-) and pCMe(-) but moderately well-separated for pCTA(-) and pCTMe(-).


The superposition strategy for arm trajectory modification in robotic manipulators.
This work deals with the problem of end-effector trajectory modification for a robot manipulator when it must respond to unexpected changes in target location. Trajectory modification and corrections are particularly important in dealing with dynamic tasks. In this paper, we present and discuss the superposition strategy derived from the study of arm trajectory modification in human subjects. According to this strategy, the motion toward the initial target location continues unmodified as planned from its beginning to its end even after the target location has unexpectedly changed. However, a trajectory leading from the first target to the final one is added vectorially to the initial one to yield the combined modified motion. A method for choosing the temporal parameters of this trajectory modification scheme is suggested so as to minimize the total travelling time under existing kinematic constraints (including both joint and hand space constraints). Then, a variant of this strategy is presented, dealing with trajectory modification in the case that the targets (both the initial and final ones) specify the desired end-point orientation rather than position.


Hemosiderin in sputum macrophages may predict infective exacerbations of chronic obstructive pulmonary disease: a retrospective observational study.
Infective exacerbations of COPD are common and are accompanied by neutrophilic bronchitis in sputum. Increased respiratory iron content has been associated with respiratory tract infection, though it is unclear if this represents a predisposing factor for infection or the sequelae of inflammation. Iron overload, as assessed in the airways, may be an important biomarker for recurrent infective exacerbations of COPD. The purpose of our study was to determine if hemosiderin in sputum macrophages is related to infective exacerbations of COPD. We undertook a retrospective observational study of 54 consecutive patients who presented with an exacerbation of COPD and had sputum examined including assessment for hemosiderin in alveolar macrophages. The relation between infective exacerbations in the previous two years and the percent of hemosiderin-positive macrophages was analyzed with linear regression. To account for the non-parametric distribution of infective exacerbations, negative binomial regression modelling was used to account for other covariates. The percent of hemosiderin positive alveolar macrophages (hemosiderin index), analyzed parametrically and non-parametrically, demonstrated a significant correlation with increasing numbers of infective exacerbations in the previous two years. In a multivariate regression analysis, hemosiderin index was an independent predictor of infective exacerbations. COPD patients with raised hemosiderin index (≥20%) had higher levels of sputum IL-6 compared to patients with lower levels (<20%). High hemosiderin index in sputum alveolar macrophages measured at the time of AECOPD may be related to the frequency of infective exacerbations of COPD.


C4d and C3d staining in biopsies of ABO- and HLA-incompatible renal allografts: correlation with histologic findings.
Biopsies of ABO-incompatible and positive crossmatch (HLA-incompatible) renal allografts were retrospectively examined to compare results of C4d and C3d staining, and the correlation between such staining and histologic findings suggestive of antibody-mediated rejection (AMR). A total of 75 biopsies (55 protocol, 17 for graft dysfunction, 3 for other indications) of 24 ABO-incompatible grafts and 244 biopsies (103 protocol, 129 for graft dysfunction, 12 for other indications) of 66 HLA-incompatible grafts were examined; all were stained for C4d and approximately 40% for C3d. In ABO-incompatible grafts, 80% of protocol biopsies and 59% performed for graft dysfunction showed C4d staining in peritubular capillaries (PTC); this staining was not correlated with neutrophil margination in PTC. In HLA-incompatible grafts, PTC C4d was present in 26% of protocol biopsies and 60% of biopsies for graft dysfunction; 92% of biopsies with >1+ (0-4+ scale), diffuse PTC C4d had > or =1+ margination and/or thrombotic microangiopathy (TMA), compared with 12% of C4d-negative biopsies. C3d was somewhat more predictive of margination than C4d in ABO-incompatible, but not HLA-incompatible, grafts. In summary, while PTC C4d deposition indicates probable AMR in biopsies of HLA-incompatible grafts, including protocol biopsies, there is no histologic evidence that C4d deposition is correlated with injury in most ABO-incompatible grafts.


Telmisartan but not valsartan increases caloric expenditure and protects against weight gain and hepatic steatosis.
The potential effects of angiotensin II receptor blockers (ARBs) on adipose tissue biology and body weight are of considerable interest, because these agents are frequently used to treat hypertension in patients who are prone to visceral obesity, the metabolic syndrome, and diabetes. In rats fed a high-fat, high-carbohydrate diet, we compared the effects of 2 ARBs, telmisartan and valsartan, on body weight, food intake, energy expenditure, fat accumulation, fat cell size, and hepatic triglyceride levels. Telmisartan, but not valsartan, promoted increases in caloric expenditure and protected against dietary-induced weight gain. In the telmisartan-treated rats, absolute food intake, but not food intake adjusted for body weight, was lower than in valsartan-treated rats or controls. Telmisartan reduced the accumulation of visceral fat and decreased adipocyte size to a much greater extent than valsartan and was also associated with a significant reduction in hepatic triglyceride levels. Moreover, telmisartan, but not valsartan, increased the expression of both nuclear-encoded and mitochondrial-encoded genes in skeletal muscle known to play important roles in mitochondrial energy metabolism. Thus, in addition to a class effect of ARBs in modulating adipocyte size, these findings raise the possibility that certain molecules, like telmisartan, may have a particularly strong impact on fat cell volume and fat accumulation, as well as distinctive effects on energy metabolism, that may help protect against dietary-induced visceral obesity and weight gain.


Age-Related Changes in Hair Shaft Protein Profiling and Genetically Variant Peptides.
Recent reports highlight possible improvements in individual identification using proteomic information from human hair evidence. These reports have stimulated investigation of parameters that affect the utility of proteomic information. In addition to variables already studied relating to processing technique and anatomic origin of hair shafts, an important variable is hair ageing. Present work focuses on the effect of age on protein profiling and analysis of genetically variant peptides (GVPs). Hair protein profiles may be affected by developmental and physiological changes with age of the donor, exposure to different environmental conditions and intrinsic processes, including during storage. First, to explore whether general trends were evident in the population at different ages, hair samples were analyzed from groups of different subjects in their 20's, 40's and 60's. No significant differences were seen as a function of age, but consistent differences were evident between European American and African American hair profiles. Second, samples collected from single individuals at different ages were analyzed. Mostly, these showed few protein expression level differences over periods of 10 years or less, but samples from subjects at 44 and 65 year intervals were distinctly different in profile. The results indicate that use of protein profiling for personal identification, if practical, would be limited to decadal time intervals. Moreover, batch effects were clearly evident in samples processed by different staff. To investigate the contribution of storage (at room temperature) in affecting the outcomes, the same proteomic digests were analyzed for GVPs. In samples stored over 10 years, GVPs were reduced in number in parallel with the yield of identified proteins and unique peptides. However, a very different picture emerged with respect to personal identification. Numbers of GVPs sufficed to distinguish individuals despite the age differences of the samples. As a practical matter, three hair samples per person provided nearly the maximal number obtained from 5 or 6 samples. The random match probability (where the log increased in proportion to the number of GVPs) reached as high as 1 in 108. The data indicate that GVP results are dependent on the single nucleotide polymorphism profile of the donor genome, where environmental/processing factors affect only the yield, and thus are consistent despite the ages of the donors and samples and batchwise effects in processing. This conclusion is critical for application to casework where the samples may be in storage for long periods and used to match samples recently collected.


Comparison of Hospital Length of Stay and Supportive Care Utilization Between Patients Treated with CPX-351 and 7+3 for Therapy-Related Acute Myeloid Leukemia or Acute Myeloid Leukemia with Myelodysplasia-Related Changes.
CPX-351 is dual-drug liposomal encapsulation of daunorubicin and cytarabine at a fixed synergistic 1:5 molar ratio. This study determined current real-world use of CPX-351 versus conventional 7+3 (cytarabine+daunorubicin) therapy and evaluated hospital length of stay (LOS) and supportive care utilization in t-AML and AML-MRC. This retrospective, observational study utilized the Premier Healthcare Database and included patients who were aged ≥18 years with t-AML or AML-MRC and treated with CPX-351 or 7+3 between August 1, 2017 and February 28, 2019. All patients treated with 7+3 were required to be eligible for CPX-351 based on its FDA-approved indication. Outcome variables were annualized and adjusted for patient, hospital, and clinical confounding factors. The primary outcome was inpatient LOS. Secondary outcomes included use of blood products and use of anti-infectives. The study included 195 qualifying patients treated with CPX-351 and 160 patients treated with 7+3 who were eligible for CPX-351. Approximately one-third of the patients treated with CPX-351 were administered therapy in a hospital-based outpatient setting, and all patients treated with 7+3 received it in the inpatient setting. The regression-adjusted annualized inpatient LOS was shorter with CPX-351 than 7+3 (mean of 183.7 vs 197.1 days, p<0.001). The difference in mean-adjusted LOS was most pronounced for t-AML, with a mean-adjusted LOS of 168.9 versus 192.5 days for CPX-351 versus 7+3, respectively (nominal p<0.001). Supportive care utilization, including the number of administrations of red blood cells, the number of administrations of platelets, and the number of days on anti-infectives, was similar between treatment groups. CPX-351 was associated with a shorter inpatient LOS than 7+3. Supportive care use, including blood products and anti-infectives, was similar for CPX-351 and 7+3. These findings suggest CPX-351 conveys resource advantages over 7+3 in patients with newly diagnosed t-AML and AML-MRC.


Safety of hydrothermally treated kernels from edible Jatropha curcas L. (Chuta) as a novel food pursuant to Regulation (EU) 2015/2283.
Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) was asked to deliver an opinion on hydrothermally treated kernels from edible Jatropha curcas (Chuta) as a novel food (NF) pursuant to Regulation (EU) 2015/2283. Although Jatropha curcas is generally considered a toxic plant due to the presence of phorbol esters (PEs), edible varieties exist in Central America. The applicant has developed a breeding programme for an edible cultivar and proposes the kernels from this cultivar as an NF as whole kernels or fragments thereof to be used as a snack or as a food ingredient. Procedures are in place to avoid commingling with non-edible kernels, with the last steps being the analytical control of PEs concentrations in all produced batches. The Panel considers that the production process of the NF is sufficiently described and that the information provided on the composition of the NF is sufficient for its characterisation. Components of the NF were tested for genotoxicity applying the standard in vitro test battery and no genotoxic concerns have been identified. In a conservative scenario for exposure to PEs from the NF, it was assumed that all kernels contain PEs at the level of detection of the analytical method. When comparing the estimated maximum exposure to PEs with a reference point from a subchronic study in pigs, a margin of exposure ≥ 900 is obtained, which is considered sufficiently large. The presence of anti-nutritional factors does not pose safety concerns as they are within the ranges found in vegetables. The Panel concludes that the NF is safe under the proposed conditions of use.


pH-Dependent Relationship between Catalytic Activity and Hydrogen Peroxide Production Shown via Characterization of a Lytic Polysaccharide Monooxygenase from Gloeophyllum trabeum.
Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that perform oxidative cleavage of recalcitrant polysaccharides. We have purified and characterized a recombinant family AA9 LPMO, LPMO9B, from Gloeophyllum trabeum (GtLPMO9B) which is active on both cellulose and xyloglucan. Activity of the enzyme was tested in the presence of three different reductants: ascorbic acid, gallic acid, and 2,3-dihydroxybenzoic acid (2,3-DHBA). Under standard aerobic conditions typically used in LPMO experiments, the first two reductants could drive LPMO catalysis whereas 2,3-DHBA could not. In agreement with the recent discovery that H2O2 can drive LPMO catalysis, we show that gradual addition of H2O2 allowed LPMO activity at very low, substoichiometric (relative to products formed) reductant concentrations. Most importantly, we found that while 2,3-DHBA is not capable of driving the LPMO reaction under standard aerobic conditions, it can do so in the presence of externally added H2O2 At alkaline pH, 2,3-DHBA is able to drive the LPMO reaction without externally added H2O2, and this ability overlaps entirely the endogenous generation of H2O2 by GtLPMO9B-catalyzed oxidation of 2,3-DHBA. These findings support the notion that H2O2 is a cosubstrate of LPMOs and provide insight into how LPMO reactions depend on, and may be controlled by, the choice of pH and reductant.IMPORTANCE Lytic polysaccharide monooxygenases promote enzymatic depolymerization of lignocellulosic materials by microorganisms due to their ability to oxidatively cleave recalcitrant polysaccharides. The properties of these copper-dependent enzymes are currently of high scientific and industrial interest. We describe a previously uncharacterized fungal LPMO and show how reductants, which are needed to prime the LPMO by reducing Cu(II) to Cu(I) and to supply electrons during catalysis, affect enzyme efficiency and stability. The results support claims that H2O2 is a natural cosubstrate for LPMOs by demonstrating that when certain reductants are used, catalysis can be driven only by H2O2 and not by O2 Furthermore, we show how auto-inactivation resulting from endogenous generation of H2O2 in the LPMO-reductant system may be prevented. Finally, we identified a reductant that leads to enzyme activation without any endogenous H2O2 generation, allowing for improved control of LPMO reactivity and providing a valuable tool for future LPMO research.


Generation of brain tumours in mice by Cre-mediated recombination of neural progenitors in situ with the tamoxifen metabolite endoxifen.
Targeted cell- or region-specific gene recombination is widely used in the functional analysis of genes implicated in development and disease. In the brain, targeted gene recombination has become a mainstream approach to study neurodegeneration or tumorigenesis. The use of the Cre-loxP system to study tumorigenesis in the adult central nervous system (CNS) can be limited, when the promoter (such as GFAP) is also transiently expressed during development, which can result in the recombination of progenies of different lineages. Engineering of transgenic mice expressing Cre recombinase fused to a mutant of the human oestrogen receptor (ER) allows the circumvention of transient developmental Cre expression by inducing recombination in the adult organism. The recombination of loxP sequences occurs only in the presence of tamoxifen. Systemic administration of tamoxifen can, however, exhibit toxicity and might also recombine unwanted cell populations if the promoter driving Cre expression is active at the time of tamoxifen administration. Here, we report that a single site-specific injection of an active derivative of tamoxifen successfully activates Cre recombinase and selectively recombines tumour suppressor genes in neural progenitor cells of the subventricular zone in mice, and we demonstrate its application in a model for the generation of intrinsic brain tumours.


Shear-induced mixing of granular materials featuring broad granule size distributions.
Granular flows during a shear-induced mixing process are studied using discrete element methods. The aim is to understand the underlying elementary mechanisms of transition from unmixed to mixed phases for a granular material featuring a broad distribution of particles, which we investigate systematically by varying the strain rate and system size. Here the strain rate varies over four orders of magnitude and the system size varies from ten thousand to more than a million granules. A strain rate-dependent transition from quasistatic to purely inertial flow is observed. At the macroscopic scale, the contact stresses drop due to the formation of shear-induced instabilities that serves as an onset of granular flows and initiates mixing between the granules. The stress-drop displays a profound system size dependence. At the granular scale, mixing dynamics are correlated with the formation of shear bands, which result in significantly different timescales of mixing, especially for those regions that are close to the system walls and the bulk. Overall, our results reveal that although the transient dynamics display a generic behavior, these have a significant finite-size effect. In contrast, macroscopic behaviors at steady states have negligible system size dependence.


Ischaemic stroke subtypes and associated risk factors: a French population based study.
There is little reliable population based information about the distribution of risk factors among the various ischaemic stroke subtypes, even though determining risk factor profiles is of major importance to develop targeted preventive strategies. The distribution of first ever ischaemic stroke subtypes was established in a prospective population based study conducted in Dijon, France (152,606 inhabitants). Cases were collected between January 2005 and December 2006, and were classified using TOAST classification. Vascular risk factors were recorded to determine a risk factor profile for each subtype. 332 patients with first ever ischaemic stroke (150 men and 182 women) were recorded. Adjusted incidence to world population was 54/100,000/year. The distribution of ischaemic stroke subtypes was as follows: 119 (35.8%) cases of large artery atherosclerosis, 89 (26.8%) small artery occlusions, 81 (24.4%) cardioembolisms and 43 (13%) other and undetermined causes. The most frequent vascular risk factor was hypertension, irrespective of the ischaemic stroke subtype, with a total prevalence of 62%. Using multivariate regression, a positive association between cardioembolism and age (OR 1.051; 95% CI 1.026 to 1.076; p<0.001) was demonstrated and between small artery occlusion and either high blood pressure (OR 1.86; 95% CI 1.06 to 3.27; p = 0.03) or hypercholesterolaemia (OR 2.23; 95% CI 1.33 to 3.76; p = 0.02). This comprehensive prospective population based study has demonstrated that vascular risk factors exhibit a particular distribution according to the ischaemic stroke subtypes. These findings, as well as the great frequency of hypertension among stroke patients, have implications for prevention strategies, the design of clinical trials and the organisation of health care services.


Age at first sex: understanding recent trends in African demographic surveys.
To describe recent trends in age at first sex in African countries, identifying and making due allowances for a variety of common reporting errors. Demographic and Health Surveys (DHS) data from six African countries conducting three or more surveys since 1985 were analysed using survival analysis techniques, combining information on virginity status and retrospective reporting of age at first sex. Hazard analysis was used to separate the effects of reporting error and compositional change and to estimate true changes in sexual debut over time. A multistate life table analysis incorporating transition to first marriage allowed cohorts to be classified according to person years spent as virgins, as sexually active unmarried, and married. Intersurvey comparisons generally suggested a slow secular rise in age at first sex. However, tracing birth cohorts between surveys revealed inconsistencies--median ages reported by female members of a birth cohort in their teens were generally higher than those reported when they reached their twenties, even when allowing for residence and education changes--probably a result of young, sexually active women denying they had ever had sex. Male birth cohorts tend to display the opposite kind of bias. Uganda, Kenya, and Ghana have experienced a more pronounced and unambiguous decline in premarital sexual activity than Tanzania, Zambia, and Zimbabwe, with statistically significant increases in age at first sex. In addition, Uganda has maintained a very short interval between onset of sexual activity and marriage for both sexes.


Identification of promoter elements responsible for gonad-specific expression of zebrafish Deadend and its application to ovarian germ cell derivation.
We discovered that a 150-bp region of zebrafish deadend (dnd) spanning the translation start codon, exon 1 and part of intron 1 is required to direct heterologous neomycin-resistance gene (neo) expression specifically in the gonad, similar to endogenous dnd. Using an 8.3-kb dnd promoter that contains this 150-bp region, we generated Tg(dnd:neo-dnd) transgenic zebrafish in which the expression of Neo was detected specifically in ovarian germ cells. The transgenic fish were used to initiate primary ovarian germ cell cultures with antibiotic G418 to select ovarian germ cells and eliminate ovarian somatic cells. RT-PCR results demonstrated that the drug-selected ovarian germ cells continued to express germ-cell markers nanos3, vasa and dnd. Growth assays demonstrated that recombinant zebrafish Lif had a significant mitogenic effect on the ovarian germ cells. When long-term ovarian germ cell cultures were transplanted into two-week-old infertile larvae, they successfully colonized and directed the formation of a truncated gonad in the recipient adult fish. Histological examination of the recipient adult fish revealed that 9 out of 34 individuals (26%) possessed donor-derived cells in their gonads. The identification of zebrafish dnd promoter and the use of this promoter to generate Tg(dnd:neo-dnd) led to the success of germ cell isolation through drug selection to generate homogenous germ cells that can be used to study zebrafish germ cell biology and may lead to a cell-mediated gene transfer strategy.


Intracerebral Injection of Extracellular Vesicles from Mesenchymal Stem Cells Exerts Reduced Aβ Plaque Burden in Early Stages of a Preclinical Model of Alzheimer's Disease.
Bone marrow Mesenchymal Stem Cells (BM-MSCs), due to their strong protective and anti-inflammatory abilities, have been widely investigated in the context of several diseases for their possible therapeutic role, based on the release of a highly proactive secretome composed of soluble factors and Extracellular Vesicles (EVs). BM-MSC-EVs, in particular, convey many of the beneficial features of parental cells, including direct and indirect β-amyloid degrading-activities, immunoregulatory and neurotrophic abilities. Therefore, EVs represent an extremely attractive tool for therapeutic purposes in neurodegenerative diseases, including Alzheimer's disease (AD). We examined the therapeutic potential of BM-MSC-EVs injected intracerebrally into the neocortex of APPswe/PS1dE9 AD mice at 3 and 5 months of age, a time window in which the cognitive behavioral phenotype is not yet detectable or has just started to appear. We demonstrate that BM-MSC-EVs are effective at reducing the Aβ plaque burden and the amount of dystrophic neurites in both the cortex and hippocampus. The presence of Neprilysin on BM-MSC-EVs, opens the possibility of a direct β-amyloid degrading action. Our results indicate a potential role for BM-MSC-EVs already in the early stages of AD, suggesting the possibility of intervening before overt clinical manifestations.


Hydrothermal growth of free standing TiO2 nanowire membranes for photocatalytic degradation of pharmaceuticals.
Highly entangled TiO(2) nanowires were directly synthesized by hydrothermal growth on Ti substrates at 180 °C utilizing various organic solvents to oxidize Ti. The growth mechanism, microstructure and phase transition of TiO(2) nanowire membranes were investigated in detail. TiO(2) nanowires, with diameters of 10-20 nm and lengths up to 100 μm, show a phase transition from Type-B to anatase by annealing at 700 °C. Robust, free standing TiO(2) nanowire membranes with millimeter level thickness can be cleaved from Ti substrates or directly prepared from thin Ti foils. These porous TiO(2) membranes, while effective for mechanical microfiltration, can also photocatalytically degrade pharmaceuticals such as trimethoprim under UV irradiation.


[The influence and significance of obtaining, storing and assay on estrogen receptor (ER) content in breast cancer tissue].
Estrogen receptor (ER) was measured in 275 samples from 170 patients with breast cancer by DCC assay. The results showed that the ER status and content of cytosol separated by ultracentrifuge (105,000 X G) or orthocentrifuge (1,200 X G) confirmed well (P greater than 0.5). Therefore, the ordinary centrifuge is suitable. But there was a poor correlation between the ER level measured by DCC method and that by Lee's cytochemical method (P greater than 0.05). The store time of the sample in liquid nitrogen did not seem to effect much on the ER positive rate (chi 2 = 0.7686). The ER status in samples obtained from the primary or metastatic lesion or at different intervals were generally similar in the same patient (P greater than 0.2). It is suggested that the ER status be an inherent property of patients with breast cancer. The authors propose that more samples be taken in order to render the determination more reliable.


Family management ofchildren who experience sickle cell disease: a qualitative study.
to know the family management experience of children with sickle cell disease in the light of the Family Management Style Framework. a qualitative case study carried out between September/2015 and July/2016 with 12 members of eight families registered in a blood center in Minas Gerais. The semi-structured interviews were recorded, and the data were analyzed and interpreted by the hybrid model thematic analysis. three management styles were identified: five families in the accommodating style; two families in the struggling style; and only one family in the enduring style.It was noted that empowerment was paramount in the acquisition of skills and abilities to care for these children. family management knowledge of children with sickle cell disease provided a reflection on nurses' role in supporting, orienting and encouraging the empowerment of these families aiming at the search for comprehensive care.


Multi-targeted therapeutic effects of Sankudiwan (SKDW) in myocardial ischemia-reperfusion injury: a comprehensive study.
This study aimed to investigate the therapeutic effects and underlying mechanisms of Sankudiwan (SKDW) on myocardial ischemia-reperfusion injury (MIRI) in a rat model. Rats were subjected to MIRI and treated with varying doses of SKDW. The myocardial infarct size, cardiac function, histological changes, apoptosis, and inflammation were assessed using TTC staining, echocardiography, Hematoxylin and Eosin (HE) staining, TUNEL staining, and ELISA assays. We further explored SKDW's influence on cardiomyocyte mitochondria and inflammatory factor expression. Moreover, oxidative stress-related parameters and differentially expressed genes were analyzed using bioinformatics approaches. SKDW significantly reduced myocardial infarct size and improved cardiac function, demonstrating a dose-dependent therapeutic potential. It ameliorated myocardial tissue damage at the histological level, inhibited cardiomyocyte apoptosis, and mitigated inflammatory response. SKDW also enhanced mitochondrial energy metabolism and suppressed the levels of oxidative stress markers. Bioinformatics analysis identified key differentially expressed genes (DEGs), including cbln1, Tgm1, Trh, and Ccl27, possibly mediating the therapeutic effects of SKDW. SKDW exerts its therapeutic effects on MIRI through the modulation of several genes and pathways related to inflammation, apoptosis, mitochondrial function, and oxidative stress. Our findings provide a scientific basis for the clinical application of SKDW in the treatment of MIRI.


Nuclear organization and transcriptional dynamics in Dictyostelium.
The Dictyostelium model has a set of features uniquely well-suited to developing our understanding of transcriptional control. The complete Dictyostelium discoideum genome sequence has revealed that many of the molecular components regulating transcription in larger eukaryotes are conserved in Dictyostelium, from transcription factors and chromatin components to the enzymes and signals that regulate them. In addition, the system permits visualization of single gene firing events in living cells, which provides a more detailed view of transcription and its relationships to cell and developmental processes. This review will bring together the available knowledge of the structure and dynamics of the Dictyostelium nucleus and discuss recent transcription imaging studies and their implications for stability and accuracy of cell decisions.


Does sigma receptor antagonism predict clinical antipsychotic efficacy?
The psychotogenic actions of sigma receptor agonists, such as pentazocine, have led to the hypothesis that sigma receptor antagonists may be putative antipsychotic agents. In this study, BW234U, a selective but relatively weak sigma receptor antagonist was compared at two different dosage ranges with chlorpromazine and placebo in a double-blind randomized treatment trial in schizophrenic patients undergoing acute exacerbation. During the 4-week blinded treatment period, there was a modest drop in Brief Psychiatric Rating Scale (BPRS) score in the chlorpromazine group, however, neither dosage range of BW234U, nor placebo produced a significant drop in the BPRS. Our results suggest that BW234U is an ineffective anti-psychotic agent in schizophrenics experiencing acute exacerbation of their illness. Due to BW234U's relatively weak antagonism at the sigma recognition site, this does not rule out the possibility that more potent and equally selective sigma antagonists may possess antipsychotic efficacy.


Arsenic hyperaccumulation by Pteris vittata and Pityrogramma calomelanos: a comparative study of uptake efficiency in arsenic-treated soils and waters.
This work comprised of the comparative study of arsenic (As) uptake efficiency by Pteris vittata and Pityrogramma calomelanos grown in (i) As amended soils (0-600 ppm) and (ii) As tainted water (40 ppb) using a new compact continuous flow phytofiltration system in a tropical greenhouse. The As hyperaccumulation efficiency was dependent on the growth medium for the two fern species. The highest level of As detected in the fronds of P. vittata was 19,300+/-190 ppm (dry weight basis) and 11,600+/-230 ppm for Pityrogramma calomelanos, after growing for 78 days in soils amended with As. In the compact continuous flow As phytofiltration system experiments, Pityrogramma calomelanos was found to perform better than P. vittata in phytofiltrating As contaminated water under waterlogged conditions. During the 167 h of phytofiltration experiment, the removal efficiency was approximately 99% and 67% for Pityrogramma calomelanos and P. vittata systems respectively, based on an initial 40 ppb As. Pityrogramma calomelanos also required a shorter acclimatization time than P. vittata under waterlogged conditions.


Binding of streptococcal antigens to muscle tissue in vitro.
Antigens extracted from cells of Streptococcus pyogenes T6 and Streptococcus mutans strains AHT, BHT, 10449, OMZ175, and K1R adsorbed to the sarcolemmal sheath of cardiac muscle cells in vitro. Similar preparations from S. salivarius, S. sanguis, Staphylococcus aureus, and Lactobacillus casei had weak or negligible tissue-binding activity. Tissue-bound bacterial antigens were detected with homologous rabbit antisera with both indirect immunofluorescence tests and an indirect radioimmunoassay. Serological cross-reactivity was observed between the tissue-binding factors of S. pyogenes and S. mutans cells but not between the bacteria and muscle tissue. In a comparative study of extraction procedures, the greatest yield of tissue-binding factors was obtained from group A streptococci by cell disruption in buffer at 4 degrees C. Hot aqueous phenol and hot water extracts were inactive. Antibodies specific for the tissue-binding factor(s) were readily adsorbed from rabbit anti-S. pyogenes serum by a preparation of isolated cytoplasmic membranes but not by a suspension of cell wall fragments. The heart-binding component of S. pyogenes cell extracts was inactivated by protease digestion and heat treatment and to a lesser extent by periodic acid oxidation. The capacity of heart cell components to adsorb streptococcal antigens was reduced by protease treatment but not by the action of neuraminidase, hyaluronidase, organic solvents, or detergents.


Antioxidant activity of some Jordanian medicinal plants used traditionally for treatment of diabetes.
Medicinal plants are being used extensively in Jordanian traditional medicinal system for the treatment of diabetes symptoms. Twenty one plant samples were collected from different Jordanian locations and used for antioxidant evaluation. The level of antioxidant activity was determined by DPPH and ABTS assays in relation to the total phenolic contents of the medically used parts. The most frequently used plant parts as medicines were fruit, shoot and leaves. The total phenolic contents of methanol and aqueous extracts, from plants parts, ranged from 6.6 to 103.0 and 3.0 to 98.6 GAE mg g(-1) of plant part dry weight, respectively. DPPH-TEAC of the methanol extracts of plants parts were varied from 4.1 to 365.0 mg g(-1) of plant dry weight versus 0.6 to 267.0 mg g(-1) in aqueous extracts. Moreover, the mean values of ABTS*- (IC50) varied from 6.9 to 400.0 microg dry weight mL(-1) ABTS in methanol extracts versus 9.8 to 580.5 microg mL(-1) in aqueous extracts. According to their antioxidant capacity, the plants were divided into three categories: high (DPPH-TEAC > or = 80 mg g(-1) ), (i.e., Punica granatum peel, Quercus calliprinos leave, Quercus calliprinos fruit, Cinchona ledgeriana and Juniperus communis leave), moderate (DPPH-TEAC range 20-80 mg g(-1)) (i.e., Salvia fruticosa shoot, Crataegus azarolus stem, Crataegus azarolus leave, Varthemia iphionoides shoot, Artemisia herba-alba shoot, Thymus capitatus shoot, Morus nigra leaves and Arum palaestinum leaves) and low antioxidant plants (DPPH-TEAC < 20 mg g(-1)), (i.e., Matricaria aurea shoot, Artemisia judaica shoot, Teucrium polium shoot, Pinus halepenss pollen grains, Sarcopoterium spinosum root, Crataegus azarolus fruit, Inula viscose shoot and Achillea fragrantissima shoot). The antioxidant activity of these plant's extracts and their potential rule in radical scavenging agreed with their potential use by Jordanian population as a traditional anti-diabetic agents.


Potential use of local and systemic humoral immune response parameters to forecast Mycoplasma hyopneumoniae associated lung lesions.
Immunopathological events are key for the development of enzootic pneumonia (EP), which is macroscopically observed as cranioventral pulmonary consolidation (CVPC). This study aimed to investigate the putative association between the humoral immune response against Mycoplasma hyopneumoniae (M. hyopneumoniae) and prevalence and extension of CVPC in 1) experimentally infected pigs, 2) slaughtered pigs and 3) sequentially necropsied pigs in a longitudinal study. CVPC was scored by means of the European Pharmacopoeia recommended methodology. Specific IgG, IgG1 and IgG2 antibodies were assessed in serum. In addition, mucosal IgG and IgA antibodies were analyzed in broncho-alveolar lavage fluid (BALF) from experimentally challenged pigs. The systemic humoral immune response in experimentally infected pigs was delayed in onset whereas humoral respiratory mucosal immune response appeared more rapidly but declined earlier. Although low, BALF IgG antibodies showed the highest correlation with CVPC scores (r = 0.49, p<0.05). In slaughter-aged pigs, both percentage of lungs with CVPC and mean lung lesion score were significantly higher in M. hyopneumoniae seropositive farms compared to the seronegative ones (p<0.001). Similarly, seropositive sequentially necropsied pigs showed more severe CVPC than seronegative ones. Overall, mean serological values might help to forecast prevalence and severity of EP-like lung lesions using a population based approach. Remarkably, the specific systemic humoral immune response was found to be predominated by the IgG2 subclass, suggesting a dominant Th1-mediated immune response to M. hyopneumoniae.


Flow-induced vasomotor response to tachycardia of the human internal mammary artery and saphenous vein grafts late following bypass surgery.
Increasing blood flow results in endothelium-dependent vasodilation of angiographically normal epicardial coronary arteries in humans. To compare the ability of internal mammary artery (IMA) and saphenous vein (SV) used as coronary artery bypass grafts to adapt their vasomotor tone to an increase in myocardial blood flow demand induced by tachycardia, 10 IMA and seven SV grafts were studied by quantitative angiography more than 6 months (range, 6-82 months) after surgery in 17 patients. All grafts were angiographically smooth and implanted to large nonakinetic perfusion zones. Angiograms were obtained in basal sinus rhythm (71 +/- 10 beats/min), during atrial pacing (129 +/- 9 beats/min), and after intragraft infusion of 1 mg isosorbide dinitrate (ISDN). In basal sinus rhythm, the cross-sectional area of SV was superior to that of IMA (10.7 +/- 3.6 versus 5.4 +/- 1.7 mm2; p less than 0.01). All IMA dilated during pacing (+14.8 +/- 11.5%; p less than 0.005) and a further increase in IMA cross-sectional area occurred after ISDN (+27.4 +/- 20.1% versus basal; p less than 0.005). In contrast, SV failed to dilate during pacing (+0.3 +/- 5.4%; p = NS) and after ISDN (+0.7 +/- 6.0% versus basal; p = NS). Thus, an increase in myocardial blood flow demand induced by tachycardia results in dilation of IMA but not of SV grafted on coronary vessels. The ability of the endothelium to modulate the vasomotor tone of IMA grafts in response to changes in blood flow could contribute to their excellent long-term functional results after coronary artery bypass surgery.


Haploid plants from pollen grains.
A method is presented by which hundreds of haploid plants of various species of Nicotiana can be raised from pollen grains. Stamens should be excised when pollen grains have been individualized, but are still uninucleate and free of starch. When grown in vitro on a relatively simple medium, some pollen grains proliferate into embryo-liké, structures which develop in stages similar to those of zygotic embryos. The plantlets mature and flower profusely, but do not set seed.


Comparative Evaluation of Antiplaque Efficacy between Essential Oils with Alcohol-Based and Chlorhexidine with Nonalcohol-Based Mouthrinses.
The aim of this study is to compare the antiplaque efficacy of alcohol-based mouthwash with essential oils and nonalcohol-based chlorhexidine mouthrinse in 4 days plaque re-formation study. The current research paper is double-blinded, parallel designed clinical trial that was conducted in the Department of Periodontics and Oral Implantology, Kalinga Institute of Dental Sciences, Bhubaneswar. This included ninety dental students of Bachelor of Dental Surgery curriculum (in the age range from 20 to 24 years) Professional cleaning with scaling and polishing was carried out to acquire a zero plaque and gingival index (GI) for all ninety patients at baseline. Ninety patients were randomly divided into three groups with thirty patients in each group named as Group A, B, and C, respectively. Two commercially available mouthrinses, one containing alcohol (labeled as Solution (A) and one without alcohol (labeled as Solution (B) were placed in identical bottles and distributed to Group A and Group B patients. Whereas, the Group C patientswere instructed to use warm saline mouth rinse. The statistical analysis was performed using the Statistical Package for the Social Sciences software (Version 18). Essential oils with an alcohol-based and chlorhexidine (CHX) alcohol-free mouthrinse (0.2%) groups compared to normal saline showed significant reductions in GI and PI scores.(P < 0.0001). The antiplaque efficacy of both alcohol-based mouthwash with essential oils and nonalcohol based CHX mouthwash were found to be equally effective in 4 days of plaque re-formation.


A genetic and transcriptomic assessment of the KTN1 gene in Parkinson's disease risk.
Parkinson's disease (PD) is a progressive neurological disorder caused by both genetic and environmental factors. An association has been described between KTN1 genetic variants and changes in its expression in the putamen and substantia nigra brain regions and an increased risk for PD. Here, we examine the link between PD susceptibility and KTN1 using individual-level genotyping data and summary statistics from the most recent genome-wide association studies (GWAS) for PD risk and age at onset from the International Parkinson's Disease Genomics Consortium (IPDGC), as well as whole-genome sequencing data from the Accelerating Medicines Partnership Parkinson's disease (AMP-PD) initiative. To investigate the potential effect of changes in KTN1 expression on PD compared to unaffected individuals, we further assess publicly available expression quantitative trait loci (eQTL) results from GTEx v8 and BRAINEAC and transcriptomics data from AMP-PD. Overall, we found no genetic associations between KTN1 and PD in our cohorts but found potential evidence of differences in mRNA expression, which needs to be further explored.


Epstein-Barr Virus-Associated Malignancies: Roles of Viral Oncoproteins in Carcinogenesis.
The Epstein-Barr virus (EBV) is the first herpesvirus identified to be associated with human cancers known to infect the majority of the world population. EBV-associated malignancies are associated with a latent form of infection, and several of the EBV-encoded latent proteins are known to mediate cellular transformation. These include six nuclear antigens and three latent membrane proteins (LMPs). In lymphoid and epithelial tumors, viral latent gene expressions have distinct pattern. In both primary and metastatic tumors, the constant expression of latent membrane protein 2A (LMP2A) at the RNA level suggests that this protein is the key player in the EBV-associated tumorigenesis. While LMP2A contributing to the malignant transformation possibly by cooperating with the aberrant host genome. This can be done in part by dysregulating signaling pathways at multiple points, notably in the cell cycle and apoptotic pathways. Recent studies also have confirmed that LMP1 and LMP2 contribute to carcinoma progression and that this may reflect the combined effects of these proteins on activation of multiple signaling pathways. This review article aims to investigate the aforementioned EBV-encoded proteins that reveal established roles in tumor formation, with a greater emphasis on the oncogenic LMPs (LMP1 and LMP2A) and their roles in dysregulating signaling pathways. It also aims to provide a quick look on the six members of the EBV nuclear antigens and their roles in dysregulating apoptosis.


[Physiopathology of sleep respiratory obstruction anomalies. Experimental methods and practical applications].
Recent progress in the physiology of the upper airways has led to significant advances in evaluating the dynamics of upper airway occlusion during sleep. Measuring the collapsibility of the upper airways and localizing the site of obstruction should theoretically lead to efficacious non-mechanical treatment and enable selection of patients susceptible of benefiting from surgical procedures. From a diagnostic point of view, fluctuations in the nasal pressure curve appear to give more precise information on ventilatory changes and their association with disrupted sleep pattern than conventional parameters such as thermistance signals or induction plethysmography. Again, from a pathophysiological point of view, little is known about the role or tissue inflammation and repeated upper airway trauma in the transition from simple snoring to sleep apnea syndrome. Current studies lead to the conclusion that the dilatroy muscles of the upper airways in the apneic subject are the target for adaptive trophic, immunohistochemical and metabolic phenomena in response to stimulation comparable to training against resistance. Although there is no evidence that sleep affects neuromuscular activity in apneic subjects, the disappearance of compensatory hyperactivity on awakening and the reduction of muscle contraction efficacy (transmission of the dilatory force to soft tissues) observed in patients may explain the greater instability of the upper airways characteristic of sleep apnea syndrome.


Isolation and characterization of casein kinase I from Dictyostelium discoideum.
In the present study, the molecular cloning and characterization of a 49-kDa form of casein kinase (CK)I from Dictyostelium discoideum is reported. The predicted amino acid sequence shares 70% identity with the catalytic domain of the mammalian delta and epsilon isoforms, Drosophila CKIepsilon and Schizosaccharomyces pombe Hhp1, and 63% identity with Hrr25, a 57-kDa form of yeast CK involved in DNA repair. D. discoideum CKI (DdCKI) was expressed in vegetative asynchronous cells as well as in differentiated cells, as detected by Northern-blot analysis. The level of DdCKI expression did not change during the cell cycle. Antibodies raised against a truncated version of the protein recognized a 49-kDa protein from D. discoideum extracts. Protein expression paralleled the pattern found for the RNA. The expression of DdCKI in Escherichia coli resulted in an active enzyme that autophosphorylated and phosphorylated casein. Immunofluorescence assays showed that DdCKI was localized in the cytoplasm and nuclei of Dictyostelium cells. The lack of disruptants of the CKI gene suggests that this protein is essential for the vegetative growth of D. discoideum. Overexpression of DdCKI resulted in cells with increased resistance to hydroxyurea, suggesting a potential role for this kinase in DNA repair.


Human bone marrow mesenchymal stem cells secrete endocannabinoids that stimulate in vitro hematopoietic stem cell migration effectively comparable to beta-adrenergic stimulation.
Granulocyte colony-stimulating factor (G-CSF) is a well-known hematopoietic stem cell (HSC)-mobilizing agent used in both allogeneic and autologous transplantation. However, a proportion of patients or healthy donors fail to mobilize a sufficient number of cells. New mobilization agents are therefore needed. Endocannabinoids (eCBs) are endogenous lipid mediators generated in the brain and peripheral tissues and activate the cannabinoid receptors CB1 and CB2. We suggest that eCBs may act as mobilizers of HSCs from the bone marrow (BM) under stress conditions as beta-adrenergic receptors (Adrβ). This study demonstrates that BM mesenchymal stem cells (MSCs) secrete anandamide (AEA) and 2-arachidonylglycerol (2-AG) and the peripheral blood (PB) and BM microenvironment contain AEA and 2-AG. 2-AG levels are significantly higher in PB of the G-CSF-treated group compared with BM plasma. BM mononuclear cells (MNCs) and CD34+ HSCs express CB1, CB2, and Adrβ subtypes. CD34+ HSCs had higher CB1 and CB2 receptor expression in G-CSF-untreated and G-CSF-treated groups compared with MSCs. MNCs but not MSCs expressed CB1 and CB2 receptors based on qRT-PCR and flow cytometry. AEA- and 2-AG-stimulated HSC migration was blocked by eCB receptor antagonists in an in vitro migration assay. In conclusion, components of the eCB system and their interaction with Adrβ subtypes were demonstrated on HSCs and MSCs of G-CSF-treated and G-CSF-untreated healthy donors in vitro, revealing that eCBs might be potential candidates to enhance or facilitate G-CSF-mediated HSC migration under stress conditions in a clinical setting.


Defective PEX gene products correlate with the protein import, biochemical abnormalities, and phenotypic heterogeneity in peroxisome biogenesis disorders.
Peroxisome biogenesis disorders (PBD) comprise three phenotypes including Zellweger syndrome (ZS) (the most severe), neonatal adrenoleucodystrophy, and infantile Refsum disease (IRD) (the most mild), and can be classified into at least 12 genetic complementation groups, which are not predictive of the phenotypes. Several pathogenic genes for PBD groups have been identified, but the relationship between the defective gene products and phenotypic heterogeneity has remained unclear. We identified a mutation in the PEX2 gene in an IRD patient with compound heterozygosity for a missense mutation and the known nonsense mutation detected in ZS patients. In transfection experiments using the peroxisome deficient CHO mutant, Z65 with a nonsense mutation in the PEX2 gene, we noted the E55K mutation had mosaic activities of peroxisomal protein import machinery and residual activities of peroxisomal functions, including dihydroxyacetone phosphate acyltransferase and beta oxidation of very long chain fatty acids. The nonsense mutation severely affects these peroxisomal functions as well as the protein import. These data suggest that allelic heterogeneity of the PEX gene affects the peroxisomal protein import and functions and regulates the clinical severity in PBD.


Study of genetic divergence among wheat genotypes through random amplified polymorphic DNA.
The degree of genetic divergence was estimated in seven wheat genotypes, six exotic genotypes and one local variety, through random amplified polymorphic DNA methodology. A total of 112 DNA fragments were generated by the 15 random primers, with an average of about 7.4 bands per primer. Among the 112, 50 fragments showed polymorphism among the seven wheat genotypes. Nei and Li's similarity matrix ranged from 86.2 to 93.0%, which indicated a narrow genetic base among the genotypes. The maximum similarity, 93.0%, was observed between 12WLRG/1-12 and WL-43. The local variety, Chenab-70, showed the lowest similarity with the exotic types. We conclude that random amplified polymorphic DNA analysis can be used for the characterization and grouping of wheat genotypes; these results will be helpful in our wheat breeding program.


The morphology of the renal microvasculature in glycerol- and gentamicin-induced acute renal failure.
To elucidate abnormalities in the renal microvasculature that could account for the functional disturbances occurring in two well-established models of acute renal failure, we gave rats a single intramuscular injection of glycerol (50%, 10 ml/kg) or daily subcutaneous doses of gentamicin (100 mg/kg/day). Afferent arteriolar diameters were determined by measuring methacrylate vascular casts with SEM. The filtration barrier was examined by both SEM and TEM. The EF area was quantitated. By 3 hr, the glycerol treatment markedly decreased PADs and DADs (PAD 19.1 to 12.0 micrometers, DAD 13.8 to 7.4 micrometers, p less than 0.05). The changes were similar for both inner and outer cortical regions. By 3 days the vasoconstriction was alleviated; however, renal failure persisted. At that time, however, EF area was decreased to 43% of normal. After 10 days of gentamicin treatment, only minimal vasoconstriction occurred in the outer cortex; however, EF area was decreased to a similar degree as observed with the 3-day glycerol-treated animals. There are two phases to glycerol-induced acute renal failure. The first phase (described as readily reversible) is characterized by intense vasoconstriction. The second phase, which is not immediately reversible, is associated with a decreased EF area. Smaller outer cortical afferent arterioles and a decreased fenestral diameter and density of the glomerular endothelium are seen only after gentamicin-induced renal failure is well established (after 10 days of treatment).


Fringe-dependent notch activation and tramtrack function are required for specification of the polar cells in Drosophila oogenesis.
During Drosophila oogenesis, each egg chamber is encapsulated through the coordinated signaling of multiple pathways, resulting in the formation of polar cells at the termini and a row of stalk cells in between each egg chamber. Notch signaling is required for specification of a precursor group containing both stalk and polar cells. Together, the Notch and JAK/STAT pathways specify the stalk cells as well as a group of prepolar cells, from within that group. The mechanism by which the polar cells differentiate from the prepolar group involves apoptosis, but the pathways which control that process are largely unknown. We now demonstrate that Notch signaling, activated by Delta and transduced by the transcription factor Tramtrack, is involved in the process of refining the prepolar cell group to two polar cells. The glycosyltransferase Fringe is expressed and required cell-autonomously in prepolar cells for this process. However, the transcription factor Mirror, which inhibits fringe expression in other tissues and stages of development, as well as Serrate, one of the two known ligands for Notch, are not required for maturation of prepolar cells. This finding suggests that Fringe is necessary for generating positional information in localizing a high-affinity interaction between Notch and its ligand Delta, even if a second ligand is not essential.


GATA-1 expression pattern can be recapitulated in living transgenic zebrafish using GFP reporter gene.
In this study, DNA constructs containing the putative zebrafish promoter sequences of GATA-1, an erythroid-specific transcription factor, and the green fluorescent protein reporter gene, were microinjected into single-cell zebrafish embryos. Erythroid-specific activity of the GATA-1 promoter was observed in living embryos during early development. Fluorescent circulating blood cells were detected in microinjected embryos 24 hours after fertilization and were still present in 2-month-old fish. Germline transgenic fish obtained from the injected founders continued to express green fluorescent protein in erythroid cells in the F1 and F2 generations. The green fluorescent protein expression patterns in transgenic fish were consistent with the pattern of GATA-1 mRNA expression detected by RNA in situ hybridization. These transgenic fish have allowed us to isolate, by fluorescence-activated cell sorting, the earliest erythroid progenitor cells from developing embryos for in vitro studies. By generating transgenic fish using constructs containing other zebrafish promoters and green fluorescent protein reporter gene, it should be possible to visualize the origin and migration of any lineage-specific progenitor cells in a living embryo.


Monitoring SARS-CoV-2 in air and on surfaces and estimating infection risk in buildings and buses on a university campus.
Evidence is needed on the presence of SARS-CoV-2 in various types of environmental samples and on the estimated transmission risks in non-healthcare settings on campus. The objective of this research was to collect data on SARS-CoV-2 viral load and to examine potential infection risks of people exposed to the virus in publicly accessible non-healthcare environments on a university campus. Air and surface samples were collected using wetted wall cyclone bioaerosol samplers and swab kits, respectively, in a longitudinal environmental surveillance program from August 2020 until April 2021 on the University of Michigan Ann Arbor campus. Quantitative rRT-PCR with primers and probes targeting gene N1 were used for SARS-CoV-2 RNA quantification. The RNA concentrations were used to estimate the probability of infection by quantitative microbial risk assessment modeling and Monte-Carlo simulation. In total, 256 air samples and 517 surface samples were collected during the study period, among which positive rates were 1.6% and 1.4%, respectively. Point-biserial correlation showed that the total case number on campus was significantly higher in weeks with positive environmental samples than in non-positive weeks (p = 0.001). The estimated probability of infection was about 1 per 100 exposures to SARS-CoV-2-laden aerosols through inhalation and as high as 1 per 100,000 exposures from contacting contaminated surfaces in simulated scenarios. Viral shedding was demonstrated by the detection of viral RNA in multiple air and surface samples on a university campus. The low overall positivity rate indicated that the risk of exposure to SARS-CoV-2 at monitored locations was low. Risk modeling results suggest that inhalation is the predominant route of exposure compared to surface contact, which emphasizes the importance of protecting individuals from airborne transmission of SARS-CoV-2 and potentially other respiratory infectious diseases. Given the reoccurring epidemics caused by highly infectious respiratory viruses in recent years, our manuscript reinforces the importance of monitoring environmental transmission by the simultaneous sampling and integration of multiple environmental surveillance matrices for modeling and risk assessment.


Division versus fusion: Dnm1p and Fzo1p antagonistically regulate mitochondrial shape.
In yeast, mitochondrial division and fusion are highly regulated during growth, mating and sporulation, yet the mechanisms controlling these activities are unknown. Using a novel screen, we isolated mutants in which mitochondria lose their normal structure, and instead form a large network of interconnected tubules. These mutants, which appear defective in mitochondrial division, all carried mutations in DNM1, a dynamin-related protein that localizes to mitochondria. We also isolated mutants containing numerous mitochondrial fragments. These mutants were defective in FZO1, a gene previously shown to be required for mitochondrial fusion. Surprisingly, we found that in dnm1 fzo1 double mutants, normal mitochondrial shape is restored. Induction of Dnm1p expression in dnm1 fzo1 cells caused rapid fragmentation of mitochondria. We propose that dnm1 mutants are defective in the mitochondrial division, an activity antagonistic to fusion. Our results thus suggest that mitochondrial shape is normally controlled by a balance between division and fusion which requires Dnm1p and Fzo1p, respectively.


[Mental health in primary care: ways to reach an integral care].
A great challenge for the primary healthcare system, implemented by the strategy called the Family Health Program, is to incorporate actions for facing situations of violence and mental health problems. This study analyzed the care delivered to 411 children between five and eleven years of age in a primary care unit in the city of São Paulo. The clinical findings were compared to a standard inventory of symptoms (CBCL). In addition, semi-structured interviews were held with pediatricians. The study shows low capacity of the pediatricians to recognize mental health problems in children. This is mainly due to deficiencies in their training and lack of possibilities for concrete intervention to face a complaint or diagnostic hypothesis. The reorganization process of primary care will need to provide specific technical support in mental health, incorporating more appropriate technologies for intervention such as a humanized approach and qualified listening. The inclusion of psychosocial aspects in the everyday practice of primary care will make it possible to broaden the concept of health and open way for an integrated approach to situations of violence related to children assisted by the primary care network of the Brazilian Health System.


Association of Central Nervous System Depression With Topical Brimonidine When Used for Hemostasis: A Serious Adverse Event.
Minor bleeding is the most common complication of dermatologic surgery. Topical brimonidine, 0.33%, gel has been reported for the use of hemostasis in dermatologic surgery. The safety profile and risk of systemic toxic effects when brimonidine is used topically for hemostasis is unknown. To determine the risk of systemic toxic effects of topical brimonidine, 0.33%, gel when used for hemostasis. In this case series from a private practice (Hollywood Dermatology), 2 patients presented for dermatologic procedures, complicated by persistent bleeding. Patients were treated with 10 g of brimonidine, 0.33%, gel applied under occlusion for hemostasis. Mental status, cardiopulmonary function. Both patients experienced deterioration of mental status, respiratory depression, and somnolence. Results from cardiac testing, laboratory workup, and imaging were negative for cardiac or neurologic etiology. Both patients improved in less than 24 hours. Topical brimonidine, 0.33%, gel can result in systemic central nervous system toxic effects when used as a hemostatic agent. At present, it is not possible to define a quantity with which brimonidine can be used safely, nor can a safe wound size be defined. We, therefore, urge against the use of topical brimonidine as a hemostatic agent until its safety is further investigated.


Molecular Characterization of Haemoglobin E.
To detect mutation in cases having haemoglobin A2 level >7% on high performance liquid chromatography. The cross-sectional, descriptive study was conducted from July 2017 to December 2018 at the Department of Haematology and Human Genetics and Molecular Biology, University of Health Sciences, Lahore, Pakistan, and comprised patients of either gender with haemoglobin A2 ≥7%. The samples were collected from different cities of Punjab in collaboration with the Punjab Thalassemia Prevention Programme, Lahore. The samples were subjected to complete blood count and high performance liquid chromatography using automated haematology analysers and variant-II beta thalassemia short programme, respectively. To analyse haemoglobin E mutations at the molecular level, polymerase chain reaction-restriction fragment length polymorphism (PCR_RFLP) was performed using a type IIS restriction endonuclease known as Mnl1 (derived from Moraxella nonliquefaciens) to cleave DNA at specific sites and the results were further confirmed on randomly selected samples using Sanger sequencing. Data was analysed using SPSS 25. Of the 39 patients, 15(38.5%) were males and 24(61.5%) were females. The overall median age was 14 (23) years. There were 29 (74.4%) patients with thalassemia family history, and 22(56.4%) had a positive family history of transfusion related to thalassemia, while no patient had a family history of iron therapy. The median haemoglobin A, haemoglobin A2 and haemoglobin F levels were 72.2 (65.2-79.1) %, 26.6 (19.1-34.0) % and 0.9 (-0.8-2.6) %, respectively. After molecular investigation, HbAE mutation was found in 23(59%) patients, while wild type HbAA genotype was found in 16(41%). The heterozygous HbE mutation was present in 23(59%) patients. Frequently missed/undiagnosed cases of haemoglobin E that co-elute with haemoglobin A2 in the same high performance liquid chromatography window were detected among those with haemoglobin A2 ≥7%.


Effects of single-pulse transcranial magnetic stimulation (TMS) on functional brain activity: a combined event-related TMS and evoked potential study.
To further evaluate the potential of slew-rate limiting amplifiers to record electrophysiological signals in spite of concurrent transcranial magnetic stimulation (TMS), and to explore the effects of single-pulse TMS on electroencephalographic (EEG) correlates of functional brain activity. Visual-evoked potentials (VEPs) to checkerboards were recorded in 7 right-handed subjects, while single-pulse TMS was applied to the occipital pole either at visual stimulus onset, during the build-up or at the expected peak of the early VEP component P1 (VIS&TMS). Timing of TMS was individually adjusted based on each subject's VEP-latency. A condition of TMS without concurrent visual stimulation (TMS(alone)) served for subtraction purposes (VIS&TMS minus TMS(alone)) to partial out TMS-related contaminations of the EEG signal. When TMS was applied at visual stimulus onset, VEPs (as calculated by subtraction) perfectly matched control VEPs to visual stimulation alone. TMS at around P1, in contrast, modified the targeted (P1) and the subsequent VEP component (N1), independently of whether TMS was given at build-up or peak. The retrieval of regular VEPs with concomitant TMS at visual stimulus onset suggests that the employed EEG system and subtraction procedure are suited for combined EEG-TMS studies. The VEP changes following TMS at around P1 provide direct clues on the temporal dynamics of TMS pulse effects on functional activity in the human brain. Our data suggest effects of relatively long duration (approximately 100 ms) when TMS is applied while functional neuronal activity evolves.


[11 years since the discovery of the human immunodeficiency virus].
The nucleotide sequence of the human immunodeficiency virus (HIV-1) gp120 variable region (V3) was obtained by sequencing the product of the polymerase chain reaction (PCR) amplification of seven HIV-1 Mexican isolates from patients who had been infected either sexually or through blood products. Mexican isolates showed a 61.4% homology between them and three viral subtypes were demonstrated. One of these corresponds to three sexually infected patients, another to two hemophiliac patients infected through blood products or transfusions and the third subtype to either sexually or blood infected patients. The latter shows a great similarity to the HTLV-IIIb/LAV prototype strain. Antigenic profiles of this region show at least three neutralization patterns for these isolates as well as one virus that does not have a high predictive value epitope in the V3 region. Our data show that Mexico has a high heterogeneity of HIV-1 circulating strains, similar to that reported in other regions of the world.


Subinhibitory Concentrations of Bacteriostatic Antibiotics Induce relA-Dependent and relA-Independent Tolerance to β-Lactams.
The nucleotide (p)ppGpp is a key regulator of bacterial metabolism, growth, stress tolerance, and virulence. During amino acid starvation, the Escherichia coli (p)ppGpp synthetase RelA is activated by deacylated tRNA in the ribosomal A-site. An increase in (p)ppGpp is believed to drive the formation of antibiotic-tolerant persister cells, prompting the development of strategies to inhibit (p)ppGpp synthesis. We show that in a biochemical system from purified E. coli components, the antibiotic thiostrepton efficiently inhibits RelA activation by the A-site tRNA. In bacterial cultures, the ribosomal inhibitors thiostrepton, chloramphenicol, and tetracycline all efficiently abolish accumulation of (p)ppGpp induced by the Ile-tRNA synthetase inhibitor mupirocin. This abolishment, however, does not reduce the persister level. In contrast, the combination of dihydrofolate reductase inhibitor trimethoprim with mupirocin, tetracycline, or chloramphenicol leads to ampicillin tolerance. The effect is independent of RelA functionality, specific to β-lactams, and not observed with the fluoroquinolone norfloxacin. These results refine our understanding of (p)ppGpp's role in antibiotic tolerance and persistence and demonstrate unexpected drug interactions that lead to tolerance to bactericidal antibiotics.


Role of the p16 tumor suppressor gene in cancer.
Since its discovery as a CDKI (cyclin-dependent kinase inhibitor) in 1993, the tumor suppressor p16 (INK4A/MTS-1/CDKN2A) has gained widespread importance in cancer. The frequent mutations and deletions of p16 in human cancer cell lines first suggested an important role for p16 in carcinogenesis. This genetic evidence for a causal role was significantly strengthened by the observation that p16 was frequently inactivated in familial melanoma kindreds. Since then, a high frequency of p16 gene alterations were observed in many primary tumors. In human neoplasms, p16 is silenced in at least three ways: homozygous deletion, methylation of the promoter, and point mutation. The first two mechanisms comprise the majority of inactivation events in most primary tumors. Additionally, the loss of p16 may be an early event in cancer progression, because deletion of at least one copy is quite high in some premalignant lesions. p16 is a major target in carcinogenesis, rivaled in frequency only by the p53 tumor-suppressor gene. Its mechanism of action as a CDKI has been elegantly elucidated and involves binding to and inactivating the cyclin D-cyclin-dependent kinase 4 (or 6) complex, and thus renders the retinoblastoma protein inactive. This effect blocks the transcription of important cell-cycle regulatory proteins and results in cell-cycle arrest. Although p16 may be involved in cell senescence, the physiologic role of p16 is still unclear. Future work will focus on studies of the upstream events that lead to p16 expression and its mechanism of regulation, and perhaps lead to better therapeutic strategies that can improve the clinical course of many lethal cancers.


Neonatal Mandibular Distraction Without a Consolidation Period: Is It Safe? Is it Effective?
Consolidation is a standard part of most post-distraction protocols. This study aims to determine whether the deliberate omission of a consolidation period following mandibular distraction (MD) in neonates with Pierre Robin sequence (PRS) affected the airway outcomes and complication rates. A retrospective chart review of 28 neonates with PRS who underwent MD for severe airway obstruction between 2009 and 2014 was performed. Neonates were split into 2 cohorts: those with a very short or no consolidation (no consolidation group, n = 18) and those with a traditional consolidation period (consolidation group, n = 10). Mean consolidation length was 25 days in the conventional consolidation group and 1.5 days in the no consolidation group (P < 0.01). Postoperatively, both groups showed equivalent improvements in their obstructive index as measured by polysomnography. Compared with the patients who underwent convention consolidation, patients in the "no consolidation" group did not require a greater number of repeat distraction, supplemental oxygen, or tracheostomy. Nor was there any significant difference in the number of complications between the 2 groups. In neonates with PRS treated with MD for severe airway obstruction, the omission of a consolidation period does not appear to significantly affect the complication rate or resolution of airway obstruction.


[Evolution of voice assessment and quality of life after endoscopic laser cordectomy].
Endoscopic laser cordectomy is a minimally invasive surgical technique for Tis, T1 and selected T2 glottic carcinoma. The purpose of the study was to evaluate prospectively pre-operatively and 3 months after surgery i) acoustic and perceptual parameters, ii) quality of life through a self evaluation (VHI) test. A prospective longitudinal evaluation of 13 male patients, shared-out onto 3 surgical procedures (cordectomy type I, grouped type II and III, and type V). Patients were recorded before and at 1 and 3 months after surgery. The parameters recorded were; acoustic parameters (F0, Jitter, Shimmer), Maximum Phonation Time (MPT), perceptual evaluation using the GRBAS scale and the self evaluation scale with the Voice Handicap Index (VHI). No statistics could be done because of the limited number of patients in each group. Perceptual and self evaluation data showed a favorable progression between 1 and 3 months for patients in group Type I, II and III cordectomy. Evolution of acoustic parameters is more difficult to analyze. The voice of the Cordectomy type V group remains whispery at 3 months after surgery. The quality of voice is a very important criterion for the patients. During the first 3 months after laser cordectomy, the perceptual vocal function and the self evaluation improves. But for the acoustic data, 3 months is probably too short to objectively quantify an improvement especially after type V cordectomy.


Curative in vivo hematopoietic stem cell gene therapy of murine thalassemia using large regulatory elements.
Recently, we demonstrated that hematopoietic stem/progenitor cell (HSPC) mobilization followed by intravenous injection of integrating, helper-dependent adenovirus HDAd5/35++ vectors resulted in efficient transduction of long-term repopulating cells and disease amelioration in mouse models after in vivo selection of transduced HSPCs. Acute innate toxicity associated with HDAd5/35++ injection was controlled by appropriate prophylaxis, making this approach feasible for clinical translation. Our ultimate goal is to use this technically simple in vivo HSPC transduction approach for gene therapy of thalassemia major or sickle cell disease. A cure of these diseases requires high expression levels of the therapeutic protein (γ- or β-globin), which is difficult to achieve with lentivirus vectors because of their genome size limitation not allowing larger regulatory elements to be accommodated. Here, we capitalized on the 35 kb insert capacity of HDAd5/35++ vectors to demonstrate that transcriptional regulatory regions of the β-globin locus with a total length of 29 kb can efficiently be transferred into HSPCs. The in vivo HSPC transduction resulted in stable γ-globin levels in erythroid cells that conferred a complete cure of murine thalassemia intermedia. Notably, this was achieved with a minimal in vivo HSPC selection regimen.


Targeting metastatic cancer from the inside: a new generation of targeted gene delivery vectors enables personalized cancer vaccination in situ.
The advent of pathotropic (disease-seeking) targeting technologies, combined with advanced gene delivery vectors, provides a unique opportunity for the systemic delivery of immunomodulatory cytokine genes to remote sites of cancer metastasis. When injected intravenously, such pathotropic nanoparticles seek out and accumulate selectively at sites of tumor invasion and neo-angiogenesis, resulting in enhanced gene delivery, and thus cytokine production, within the tumor nodules. Used in conjunction with a primary tumoricidal agent (e.g., Rexin-G) that exposes tumor neoantigens, the tumor-targeted immunotherapy vector is intended to promote the recruitment and activation of host immune cells into the metastastic site(s), thereby initiating cancer immunization in situ. In this study, we examine the feasibility of cytokine gene delivery to cancerous lesions in vivo using intravenously administered pathotropically targeted nanoparticles bearing the gene encoding granulocyte/macrophage colony-stimulating factor (GM-CSF; i.e., Reximmune-C). In vitro, transduction of target cancer cells with Reximmune-C resulted in the quantitative production of bioactive and immunoreactive GM-CSF protein. In tumor-bearing nude mice, intravenous infusions of Reximmune-C-induced GM-CSF production by transduced cancer cells and paracrine secretion of the cytokine within the tumor nodules, which promoted the recruitment of host mononuclear cells, including CD40+ B cells and CD86+ dendritic cells, into the tumors. With the first proofs of principle established in preclinical studies, we generated an optimized vector configuration for use in advanced clinical trial designs, and extended the feasibility studies to the clinic. Targeted delivery and localized expression of the GM-CSF transgene was confirmed in a patient with metastatic cancer, as was the recruitment of significant tumor-infiltrating lymphocytes (TILs). Taken together, these studies provide the first demonstrations of cytokine gene delivery to cancerous lesions following intravenous administration and extend the applications of cancer immunization in vivo.


Relaxation matching techniques-a comparison.
Many different relaxation schemes have been proposed for image analysis tasks. We have developed a general matching procedure for comparing semantic network descriptions of images, and we have implemented a variety of relaxation techniques. An automatic segmentation and description system is used to produce the image representations so that the matching procedures must cope with variations in feature values, missing objects, and possible multiple matches. This environment is used to test different relaxation matching schemes under a variety of conditions. The best performance (of those we compared), in terms of the number of iterations and the number of errors, is for the gradient-based optimization approach of Faugeras and Price. The related optimization approach of Hummel and Zucker performed almost as well, with differences primarily in difficult matches (i.e., where much of the evidence is against the match, for instance, poor segmentations). The product combination rule proposed by Peleg was extremely fast, indeed, too fast to work when global context is needed. The classical Rosenfeld, Hummel, and Zucker method is included for historical comparisons and performed only adequately, producing fewer correct matches and taking more iterations.


Bioengineered embryoids mimic post-implantation development in vitro.
The difficulty of studying post-implantation development in mammals has sparked a flurry of activity to develop in vitro models, termed embryoids, based on self-organizing pluripotent stem cells. Previous approaches to derive embryoids either lack the physiological morphology and signaling interactions, or are unconducive to model post-gastrulation development. Here, we report a bioengineering-inspired approach aimed at addressing this gap. We employ a high-throughput cell aggregation approach to simultaneously coax mouse embryonic stem cells into hundreds of uniform epiblast-like aggregates in a solid matrix-free manner. When co-cultured with mouse trophoblast stem cell aggregates, the resulting hybrid structures initiate gastrulation-like events and undergo axial morphogenesis to yield structures, termed EpiTS embryoids, with a pronounced anterior development, including brain-like regions. We identify the presence of an epithelium in EPI aggregates as the major determinant for the axial morphogenesis and anterior development seen in EpiTS embryoids. Our results demonstrate the potential of EpiTS embryoids to study peri-gastrulation development in vitro.


Complete Genome Sequence of Klebsiella aerogenes Siphophage Solomon.
Klebsiella aerogenes is a bacterium that can cause a variety of infections. Phage-based biotechnologies may be useful for controlling antibiotic-resistant strains of this bacterium. The characterization of K. aerogenes phage Solomon is described here. Solomon has a 51,775-bp genome, with structural components closely resembling those of Escherichia coli siphophage T1.


Combined gas well hydrate prevention and control technology and its application.
The high pressure in some gas wells, such as those in the Xushen gas field in Daqing, China, makes them susceptible to freezing and hydrate blockages. Downhole throttling technology is widely used to reduce costs during well construction, however, due to the limitations of temperature, pressure and depth structure, this technology is sometime applied independently in some gas wells in which freezing and blockages are a frequent problem that can seriously affect production capacity. Moreover, artificial alcohol injection of 'passive plugging' to prevent hydrate formation not only consumes significant amounts of methanol but its efficiency is also dependent on factors such as weather, personnel and equipment, so it is not a continuous solution. In order to solve the above problems, the mechanism of hydrate formation was analyzed in this study, from which a combined mechanical and chemical hydrate control process was developed. OLGA software was used to design the process parameters of the novel mechanical and chemical inhibition technology for hydrate prevention and control, and also to simulate and analyze the wellhead temperature, pressure and hydrate generation once the process was implemented. Based on the results of the parameters calculation, the downhole throttle and hydrate inhibitor automatic filling device are used to realize the functions of downhole throttle depressurization and hydrate inhibitor continuous filling, reduce the wellhead pressure and hydrate generation temperature, and ensure the continuous production of gas well. This novel combination process was subsequently tested in three wells in the Daqing gas oilfield. Measurements showed that the average daily gas increase from a single well was 0.5×104m3, methanol consumption was reduced from the original maximum daily amount of 1750 kg to just 60 kg, the manual maintenance workload was reduced by 80%, and the rate of the well openings was increased from 45% to 100%. These results proved that this technology is feasible and efficient for applications in gas wells with high downhole pressure and low wellhead temperature, and, thus, provides important technical support for the prevention of gas hydrate and improvement of gas well production.


Venom-derived peptides inhibiting Kir channels: Past, present, and future.
Inwardly rectifying K+ (Kir) channels play a significant role in vertebrate and invertebrate biology by regulating the movement of K+ ions involved in membrane transport and excitability. Yet unlike other ion channels including their ancestral K+-selective homologs, there are very few venom toxins known to target and inhibit Kir channels with the potency and selectivity found for the Ca2+-activated and voltage-gated K+ channel families. It is unclear whether this is simply due to a lack of discovery, or instead a consequence of the evolutionary processes that drive the development of venom components towards their targets based on a collective efficacy to 1) elicit pain for defensive purposes, 2) promote paralysis for prey capture, or 3) facilitate delivery of venom components into the circulation. The past two decades of venom screening has yielded three venom peptides with inhibitory activity towards mammalian Kir channels, including the discovery of tertiapin, a high-affinity pore blocker from the venom of the European honey bee Apis mellifera. Venomics and structure-based computational approaches represent exciting new frontiers for venom peptide development, where re-engineering peptide 'scaffolds' such as tertiapin may aid in the quest to expand the palette of potent and selective Kir channel blockers for future research and potentially new therapeutics. This article is part of the Special Issue entitled 'Venom-derived Peptides as Pharmacological Tools.'


Attitudes and behaviors of practicing community pharmacists towards patient counselling.
The present study was conducted to assess the attitudes and behaviors of practicing community pharmacists towards patient counselling and use of patient information leaflets in the state of Karnataka. Convenient sampling method was adopted to collect the responses with the help of self-completion questionnaires. A total of 258 practicing community pharmacists in the age group of 22-60 y of both gender with practicing experience of 2-30 y participated in the study. Majority of respondents (80%) agreed that, patient counselling is their professional obligation. About 17% of the respondents mentioned that, they try to give basic information regarding drug usage to the patient. The reasons stated by the pharmacists to provide patient counselling were, professional satisfaction (43%), patients go with satisfaction (32%), observed increase in sales (8%), and also improved patient compliance (7.5%). The major barriers for offering patient counselling were mentioned as pharmacists' inadequate knowledge and confidence (78%), doctor dispensing (72%), no professional fee (56%), poor response from patients (82%), inadequate continuous professional development programs (75%). Many respondents agreed that, patient information leaflets certainly help in counselling but available information leaflets are company generated and prescriber focused. Many respondents found the present continuing professional development module was useful and are interested in weekend workshops to update their professional knowledge (83%). Restrictions on doctor dispensing, legalization of patient counselling, regular continuing professional development programs are the factors observed to motivate the pharmacists to offer patient counselling.


Stratifying future stroke risk with incidentally-discovered white matter disease severity and covert brain infarct site.
Covert cerebrovascular disease (CCD) includes white matter disease (WMD) and covert brain infarction (CBI). Incidentally-discovered CCD is associated with increased risk of subsequent symptomatic stroke. However, it is unknown whether the severity of WMD or the location of CBI predicts risk. To examine the association of incidentally-discovered WMD severity and CBI location with risk of subsequent symptomatic stroke. This retrospective cohort study includes patients 50 years old in the Kaiser Permanente Southern California health system who received neuroimaging for a non-stroke indication between 2009-2019. Incidental CBI and WMD were identified via natural language processing of the neuroimage report, and WMD severity was classified into grades. 261,960 patients received neuroimaging; 78,555 (30.0%) were identified to have incidental WMD, and 12,857 (4.9%) to have incidental CBI. Increasing WMD severity is associated with increased incidence rate of future stroke. However, the stroke incidence rate in CT-identified WMD is higher at each level of severity compared to rates in MRI-identified WMD. Patients with mild WMD via CT have a stroke incidence rate of 24.9 per 1,000 person-years, similar to that of patients with severe WMD via MRI. Among incidentally-discovered CBI patients with a determined CBI location, 97.9% are subcortical rather than cortical infarcts. CBI confers a similar risk of future stroke, whether cortical or subcortical, or whether MRI- or CT-detected. Increasing severity of incidental WMD is associated with an increased risk of future symptomatic stroke, dependent on the imaging modality. Subcortical and cortical CBI conferred similar risks.


Immediate nonfunctional loading of implants placed simultaneously using computer-guided flapless maxillary crestal sinus augmentation with bone morphogenetic protein-2/collagen matrix.
Immediate loading has shown positive results for total, partial, or single edentulism. The effects of BMP-2 addition to graft materials on bone formation and implant stability in the early stages of healing have rarely been studied, especially in patients with severely atrophic maxillae. To evaluate the effects of simultaneously placed immediate non-functional loaded implants and bone morphogenetic protein-2 (BMP-2)-loaded Bio-Oss collagen, on bone formation and implant stability during the early healing stages of patients with a severely atrophic posterior maxilla using crestal approach. Thirty-three cases presenting posterior maxillary residual alveolar bone height of 1-3 mm were evaluated. Flapless crestal sinus augmentation surgery was performed using BMP-2-loaded Bio-Oss collagen, with non-functional implants immediately loaded after surgery. The bone height was assessed using preoperative and postoperative cone beam computed tomography (CBCT). Bone density of the sinus graft sites and implant stability (after 3 months) were evaluated using postoperative CBCT scans and Periotest, respectively. The periodontal parameters and marginal bone loss around the implant were checked after 37.3 months of final prosthesis. The survival rate of the implants was 100% and the gingiva around the implants remained healthy. All implants remained integrated, and all sinus grafts showed radiographic bone formation. The results indicated high level of bone density and good implant stability, showing minimal marginal bone loss after 37.3 months. This technique could be used in the posterior maxillary region exhibiting poor bone quantity.


Deteriorating microbiomes in agriculture - the unintended effects of pesticides on microbial life.
There is emerging concern regarding the unintentional and often unrecognized antimicrobial properties of "non-antimicrobial" pesticides. This includes insecticides, herbicides, and fungicides commonly used in agriculture that are known to produce broad ranging, off-target effects on beneficial wildlife, even at seemingly non-toxic low dose exposures. Notably, these obscure adverse interactions may be related to host-associated microbiome damage occurring from antimicrobial effects, rather than the presumed toxic effects of pesticides on host tissue. Here, we critically review the literature on this topic as it pertains to the rhizosphere microbiome of crop plants and gut microbiome of pollinator insects (namely managed populations of the western honey bee, Apis mellifera), since both are frequent recipients of chronic pesticide exposure. Clear linkages between pesticide mode of action and host-specific microbiome functionalities are identified in relation to potential antimicrobial risks. For example, inherent differences in nitrogen metabolism of plant- and insect-associated microbiomes may dictate whether neonicotinoid-based insecticides ultimately exert antimicrobial activities or not. Several other context-dependent scenarios are discussed. In addition to direct effects (e.g., microbicidal action of the parent compound or breakdown metabolites), pesticides may indirectly alter the trajectory of host-microbiome coevolution in honey bees via modulation of social behaviours and the insect gut-brain axis - conceivably with consequences on plant-pollinator symbiosis as well. In summary, current evidence suggests: (1) immediate action is needed by regulatory authorities in amending safety assessments for "non-antimicrobial" pesticides; and (2) that the development of host-free microbiome model systems could be useful for rapidly screening pesticides against functionally distinct microbial catalogues of interest.