Applying Multimodal Mass Spectrometry to Image Tumors Undergoing Ferroptosis Following In Vivo Treatment with a Ferroptosis Inducer.
Epithelial ovarian cancer (EOC) is the most common form of ovarian cancer. The poor prognosis generally associated with this disease has led to the search for improved therapies such as ferroptosis-inducing agents. Ferroptosis is a form of regulated cell death that is dependent on iron and is characterized by lipid peroxidation. Precise mapping of lipids and iron within tumors exposed to ferroptosis-inducing agents may provide insight into processes of ferroptosis in vivo and ultimately assist in the optimal deployment of ferroptosis inducers in cancer therapy. In this work, we present a method for combining matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) with secondary ion mass spectrometry (SIMS) to analyze changes in spatial lipidomics and metal composition, respectively, in ovarian tumors following exposure to a ferroptosis inducer. Tumors were obtained by injecting human ovarian cancer tumor-initiating cells into mice, followed by treatment with the ferroptosis inducer erastin. SIMS imaging detected iron accumulation in the tumor tissue, and sequential MALDI-MS imaging of the same tissue section displayed two chemically distinct regions of lipids. One region was associated with the iron-rich area detected with SIMS, and the other region encompassed the remainder of the tissue section. Bulk lipidomics confirmed the lipid assignments putatively assigned from the MALDI-MS data. Overall, we demonstrate the ability of multimodal MSI to identify the spatial locations of iron and lipids in the same tissue section and associate these regions with clinical pathology.


The co-occurrence of epilepsy and autism: A systematic review.
We aimed to review the literature to determine the incidence and prevalence of autism in epilepsy and epilepsy in autism, conditions that are often comorbid. We adhered to the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) standards, and the protocol was registered with PROSPERO. MEDLINE, Embase, PsycINFO, and the Cochrane Database of Systematic Reviews were searched from inception until July 4, 2016. Studies were included if they reported an incidence or prevalence of autism in epilepsy or epilepsy in autism. These estimates were described using mean, standard deviation, median, and interquartile range. Seventy-four studies reporting on 283,549 patients were included. The median overall period prevalence of epilepsy in people with autism was 12.1% while the median overall period prevalence of autism in people with epilepsy was 9.0% when including all population types. When excluding studies that investigated patients with syndromic epilepsy or developmental delay, the median overall period prevalence of epilepsy in people with autism was 11.2% while the median overall period prevalence of autism in people with epilepsy was 8.1%. We observed trends for sex as the prevalence of autism in epilepsy was higher in males while the prevalence of epilepsy in autism was higher in females. It is important to interpret these estimates with caution, as there was significant heterogeneity between studies. Meta-regression found no association between study quality and prevalence or incidence estimates (all p-values > 0.05). The period prevalence of epilepsy in people with autism, and vice versa, was consistently higher than previously reported estimates of the occurrence of these disorders in the general population. These findings highlight the importance of screening for autism in people who have epilepsy and epilepsy in people who have autism and may help shed light on shared pathogenesis between these conditions.


Trends and consequences of surgical conversion in the United States.
The aim of this study was to identify national utilization trends of robotic surgery for elective colectomy, conversion rates over time, and the specific impact of conversion on postoperative morbidity. Conversion to open represents a hard endpoint for minimally invasive surgery (MIS) and is associated with worse outcomes when compared to MIS or even traditional open procedures. All adult patients who underwent either laparoscopic or robotic elective colectomy from 2013 to 2018 as reported in the American College of Surgeons Quality Improvement Program (ACS-NSQIP) database were included. National trends of both robotic utilization and conversion rates were analyzed, overall and according to underlying disease (benign disease, inflammatory bowel disease (IBD), cancer), or the presence of obesity (body mass index (BMI) ≥ 30 kg/m2). Demographic and surgical risk factors for surgical conversion to open were identified through multivariable regression analysis. Further assessed were overall and specific postoperative 30-day complications, which were risk adjusted and compared between converted patients and the remaining cohort. Of 66,652 included procedures, 5353 (8.0%) were converted to open. Conversion rates were 8.5% for laparoscopic and 4.9% for robotic surgery (p < 0.0001). A decline in conversion rates over the 6-year inclusion period was observed overall and for patients with obesity. This trend paralleled an increased utilization of the robotic platform. Several surrogates for advanced disease stages for cancer, diverticulitis, and IBD and prolonged surgical duration were identified as independent risk factors for unplanned conversion, while robotic approach was an independent protective factor (OR 0.44, p < 0.0001). Patients who had unplanned conversion were more likely to experience postoperative complications (OR 2.36; 95% CI [2.21-2.51]), length of hospital stay ≥ 6 days (OR 2.86; 95% CI [2.67-3.05], and 30-day mortality (OR 2.28; 95% CI [1.72-3.02]). This nationwide study identified a decreasing trend in conversion rates over the 6-year inclusion period, both overall and in patients with obesity, paralleling increased utilization of the robotic platform. Unplanned conversion to open was associated with a higher risk of postoperative complications.


Recovery of infectious human parainfluenza virus type 3 from cDNA.
Infectious HPIV3 was produced by the intracellular coexpression of four plasmid-borne cDNAs. These separately encoded a complete HPIV3 genome (negative-sense), the HPIV3 nucleocapsid protein N, the phosphoprotein P, and the polymerase protein L. The cDNA-encoded HPIV3 genome differed from the JS wildtype (wt) strain of HPIV3 used in its construction by seven point mutations: four of these are silent mutations in the HN or L gene coding regions that serve as markers of a cDNA-derived virus, two were introduced to create an amino acid substitution that ablates an epitope recognized by the HN-specific monoclonal neutralizing antibody 423/6, and the remaining point mutation results in an incidental amino acid substitution in the HN protein at amino acid position 263. The four plasmids were transfected into HEp-2 cell monolayers and their expression was driven by T7 RNA polymerase supplied by a vaccinia virus recombinant. The titer of virus present in the harvested transfection supernatant was low (<5 PFU/ml), and the recovered recombinant virus (rJS) retained each of the seven mutations present in the cDNA from which it was derived. Despite the introduced and incidental mutations, rJS retained the wt phenotypes as regards replication at elevated temperature in vitro and efficient replication in the upper and lower respiratory tract of hamsters. rJS was also recovered from a cDNA encoding a complete antigenome (positive-sense) with slightly greater efficiency than from the negative-sense construct. The ability to generate infectious HPIV3 from cDNA should greatly enhance our ability to develop new live-attenuated parainfluenza virus vaccines, including chimeric PIV1 and PIV2 vaccines, and to understand the genetic basis of attenuation of PIV3 candidate vaccines.


Feasibility of Exploiting Physiological and Motion Features for Camera-based Sleep Staging: A Clinical Study.
Camera-based sleep monitoring is an emergent research topic in sleep medicine. The feasibility of using both the physiological features and motion features measured by a video camera for sleep staging was not thoroughly investigated. In this paper, we built a camera-based non-contact sleep monitoring setup in the Institute of Respiratory Diseases, Shenzhen People's Hospital, and created a clinical sleep dataset (nocturnal video data of 11 adults) including the expert-corrected PSG references synchronized with the video. The camera-based measurements have shown high correlations with the PSG. It obtains an overall Mean Absolute Error (MAE) of 1.5 bpm for heart-rate (HR), 0.7 bpm for breathing-rate (BR), 13.9 ms for heart-rate variability (HRV), and an accuracy of 93.5% for leg motion detection. The statistical analysis indicates that the averaged HR and variations of BR are distinct features for annotating four sleep stages (awake, REM, light sleep, and deep sleep). HRV parameter (SDNN) can clearly differentiate rapid-eye-movement (REM) and non-REM, while the leg movement is a distinctive feature for separating awake and sleep. The clinical trial demonstrated the feasibility of using physiological and motion features measured by a camera for joint sleep staging, and provides insights for sleep-related feature selection.


Online training of behavioral health providers for primary care practice.
Introduction: Many behavioral health providers have not received training in primary care practice during their education. Since 2007, the online Certificate in Primary Care Behavioral Health course has been completed by thousands of behavioral health providers. An evaluation of the course assessed whether learner's baseline confidence in their abilities to address behavioral health concerns in primary care settings changed over time, whether learning outcomes for live offerings of the course differed from asynchronous offerings, and whether learning outcomes for psychologists and social workers differed. Method: Learners were asked to rate their confidence in their abilities using 10 retrospective pre-post questions. Responses from 14 cohorts of learners, between 2011 and 2019 were assessed. T-tests and analyses of variance were used to compare groups. Results: Learners' baseline confidence in their abilities changed in three of the areas assessed. Those completing the course asynchronously reported outcomes equal to or greater than those completing the course synchronously. In all but one domain, psychologists and social workers reported equal increases in their confidence. Discussion: Learners reported significant improvements in confidence in their ability to work as behavioral health providers in primary care. With one exception, these findings did not differ for psychologists and social workers. Over time, baseline confidence of behavioral health providers enrolling in the course increased in some areas, particularly those focused on patients with substance use disorders. Learning outcomes for the asynchronous version of the course support the continued delivery of asynchronous training of behavioral health providers working in primary care. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Pulmonary hemodynamic response to exercise in subjects with prior high-altitude pulmonary edema.
Individuals with a prior history of (susceptible to high altitude pulmonary edema (HAPE-S) have high resting pulmonary arterial pressures, but little data are available on their vascular response to exercise. We studied the pulmonary vascular response to exercise in seven HAPE-S and nine control subjects at sea level and at 3,810 m altitude. At each location, both normoxic (inspired PO2 = 148 Torr) and hypoxic (inspired PO2 = 91 Torr) studies were conducted. Pulmonary hemodynamic measurements included pulmonary arterial and pulmonary arterial occlusion pressures. A multiple regression analysis demonstrated that the pulmonary arterial pressure reactivity to exercise was significantly greater in the HAPE-S group. This reactivity was not influenced by altitude or oxygenation, implying that the response was intrinsic to the pulmonary circulation. Pulmonary arterial occlusion pressure reactivity to exercise was also greater in the HAPE-S group, increasing with altitude but independent of oxygenation. These findings suggest an augmented flow-dependent pulmonary vasoconstriction and/or a reduced vascular cross-sectional area in HAPE-S subjects.


Role of DNA protection and repair in resistance of Bacillus subtilis spores to ultrahigh shock pressures simulating hypervelocity impacts.
Impact-induced ejections of rocks from planetary surfaces are frequent events in the early history of the terrestrial planets and have been considered as a possible first step in the potential interplanetary transfer of microorganisms. Spores of Bacillus subtilis were used as a model system to study the effects of a simulated impact-caused ejection on rock-colonizing microorganisms using a high-explosive plane wave setup. Embedded in different types of rock material, spores were subjected to extremely high shock pressures (5 to 50 GPa) lasting for fractions of microseconds to seconds. Nearly exponential pressure response curves were obtained for spore survival and linear dependency for the induction of sporulation-defective mutants. Spores of strains defective in major small, acid-soluble spore proteins (SASP) (alpha/beta-type SASP) that largely protect the spore DNA and spores of strains deficient in nonhomologous-end-joining DNA repair were significantly more sensitive to the applied shock pressure than were wild-type spores. These results indicate that DNA may be the sensitive target of spores exposed to ultrahigh shock pressures. To assess the nature of the critical physical parameter responsible for spore inactivation by ultrahigh shock pressures, the resulting peak temperature was varied by lowering the preshock temperature, changing the rock composition and porosity, or increasing the water content of the samples. Increased peak temperatures led to increased spore inactivation and reduced mutation rates. The data suggested that besides the potential mechanical stress exerted by the shock pressure, the accompanying high peak temperatures were a critical stress parameter that spores had to cope with.


Estrogen inhibits interleukin-18 mRNA expression in the mouse uterus.
Interleukin-18 (IL-18) is a proinflammatory cytokine expressed in female reproductive organs in humans, rats and mice. The physiological roles of uterine IL-18 and the regulatory mechanisms of IL-18 gene expression are unclear. The present study aimed to clarify the effects of estradiol-17beta (E2) and progesterone (P4) on IL-18 mRNA expression in the mouse uterus. Distribution and expression levels of IL-18 mRNA were studied using an RNase protection assay. Expression of IL-18 mRNA was observed in all organs studied, including testes, ovaries and uteri. The uterine IL-18 mRNA level of estrous mice was higher than that of diestrous mice. E2 treatment (1, 5, 25 or 250 ng/mouse) decreased uterine IL-18 mRNA levels in ovariectomized mice dose-dependently. E2 treatment acutely decreased IL-18 mRNA levels 3 h after injection, but these levels returned to the initial level after 48 h. P4 treatment (1 mg/mouse) decreased uterine IL-18 mRNA levels after 12 h, but levels returned to the initial level after 48 h. Both uterine IL-18 and IL-18Ralpha mRNAs were detected in cultured endometrial epithelial and stromal cells. These results suggest that uterine IL-18 expression is reduced by sex steroid hormones and that IL-18 acts on endometrial cells in a paracrine or autocrine manner.


The role of social support and family relationships in women's responses to battering.
For women who are in abusive relationships, social support and close family relationships may be critical to the successful resolution of the abuse. Efforts to enhance battered women's support must be predicated on an understanding of women's perceptions of effective support and the constraints they experience to seeking support. In this paper we present findings from a qualitative analysis of semistructured interviews with 31 women who were in abusive relationships. The women were interviewed three times over two-and-a-half years. Results of the analysis included that women used female friends for support more often than family members, and that women were constrained from seeking support by (a) cultural and societal sanctions against leaving the relationship; (b) a pattern of caution in relating to others or forging new relationships; and (c) forced isolation/seeing self as isolative. Relationships with family members, especially parents, were not consistently seen as useful sources of support.


Effects of drugs on immune responsiveness in rabbits.
The effects of chemotherapeutica: 6 mercaptopurine (6 MP), nitrogen mustard (HN2) and cyclophosphamide (Cy) on T and B cells were studied in standardized experimental systems. From the histological and immunological results it was concluded that: (a) A course of 9 daily 6 MP injections affected antibody production only when given after antigenic stimulation. Histologically figures of mitotic death were found among proliferating cells of plasmacellular-germinal centre-and specific cellular reactions. It was concluded that a net mitotic death of 50% would account for the observed immunological and histological effects; (b) HN2 and Cy affected antibody production itself when administered during a very restricted period (1 through 3 days after ag.). Histologically only morphological changes were found in differentiating immature plasmacells and interphase death of cells involved in germinal centre reactions. Loss of responsiveness depended on the degree of interphase death of follicular elements. Cy interfered with the formation of T helper cells in thymus-dependent antibody production; (c) L-Asparaginase (1,000 U) only prevented the origination of T-helper cells. Neither antibody-producing cells nor their precursors were affected.


Investigations of calsequestrin as a target for anthracyclines: comparison of functional effects of daunorubicin, daunorubicinol, and trifluoperazine.
Anthracycline therapy is associated with a life-threatening but poorly understood cardiotoxicity. Effects of treatment are consistent with drug-induced disruption of cardiac sarcoplasmic reticulum (SR) calcium homeostasis, including inhibition of calcium release by anthracyclines. This effect, which depends on luminal SR calcium concentration, is hypothesized to involve interactions of anthracyclines with the calcium binding protein calsequestrin (CSQ). This study was designed to test the hypothesis that an interaction between CSQ and anthracyclines could be related to alterations in SR calcium release and cardiac function. The effects of the anthracycline, daunorubicin, and its metabolite daunorubicinol were compared with those of a known CSQ inhibitor, trifluoperazine (TFP). Protein fluorescence quenching studies demonstrated that TFP, daunorubicin, and daunorubicinol bind to CSQ with apparent binding affinities in the low micromolar range. The presence of calcium decreases the drug-dependent fluorescence quenching, probably because of calcium-induced CSQ conformational changes. TFP also inhibited SR calcium release. Although the TFP IC50 value is somewhat larger than for anthracyclines, the TFP effect is also dependent on luminal SR calcium concentration. In a muscle preparation, micromolar TFP decreased cardiac contractility in a manner that implicates the involvement of SR calcium and resembles the effects of anthracyclines. These data are consistent with a mechanism in which TFP or anthracyclines impair SR calcium release and cardiac function through a mechanism involving disruption of CSQ function. Such a mechanism may contribute to anthracycline cardiotoxicity.


Assessment of IgH PCR strategies in multiple myeloma.
To compare the ability of four commonly used PCR techniques to demonstrate clonal IgH rearrangements in multiple myeloma. Bone marrow samples (containing a minimum of 10% plasma cells) were obtained from 127 patients with confirmed multiple myeloma. Framework 3 (Fr3) PCR was performed in all cases and the Framework 1 (Fr1f) PCR, which utilises six VH family specific primers, in 98 cases. In addition, 44 cases were assessed by Fr3, Fr1f, Framework 2 (Fr2) and Framework 1 consensus (Fr1 con) PCR techniques. JH primer selection was also assessed such that each PCR strategy was performed twice in each of the 44 cases, using the JH consensus primer (JH con) alone and then repeated with an equimolar mixture of JH con, JH3 and JH6 (JH mix). Clonal rearrangements were demonstrated in 71 (56%) of 127 cases with the Fr3 PCR and in 52 (53%) of 98 with the Fr1f PCR. However, by using both techniques it was possible to demonstrate clonal IgH rearrangements in 92 (75%) of 122 cases. Forty four cases were assessed by all four PCR techniques; in these cases the Fr3 and Fr1f PCRs demonstrated clonal rearrangements in 26 (59%) cases with a combined yield of 34 (77%). The Fr2 and Fr1 con PCR techniques had inferior pick up rates, demonstrating clonal rearrangements in 21 (48%) of 44 cases and a combined yield of 28 (63%). The Fr2 PCR did, however, demonstrate a clonal rearrangement in one case negative by both Fr3 and Fr1f. Two additional rearrangements were demonstrated by using JH mix; one became positive by Fr3, Fr1f and Fr2 and the other positive by Fr1f, Fr1 con and Fr2. By utilising both the Fr3 and Fr1f PCR techniques it is possible to demonstrate definitive clonal rearrangements in the majority of patients with multiple myeloma. The Fr1 con and Fr2 PCR techniques have inferior pick up rates but may detect some additional rearrangements.


[An alchemist or swindler? The case of Zbigniew Dunikowski].
In early 1930s the newspapers and street journals in Europe and the United States were frequently reporting on a case of Zbigniew Dunikowski, a Polish engineer, who claimed to be in possession of a secret formula allowing production of gold from ordinary sand and rocks. He believed that most of those materials contain some particles of gold. For the precious metal however, it takes millions of years to precipitate into the ledges that could be mined. His method was based on a conviction, that the process can be accelerated. Although he was nicknamed "Polish alchemist" very soon, his vain promises attracted attention of financiers and even some European political leaders. After few years of futile experiments, he was sued by his impatient financial backers, and arrested. While in detention, he was allowed to make the last attempt to produce gold and regain his repute and freedom. When this attempt failed, the judge sentenced him for two years in prison and ordered him to repay some 3 million francs ($100,000) to his investors. He was also fined with ... 100 francs fine (some 4 dollars). It can not be definitively stated, whether Dunikowski was truly convicted that his formula for making gold could have been working or he acted as a swindler from the very beginning. He exclaimed that the accusation of fraud was caused by bankers, who would never let his method to undermine the status quo of world's economy. The experiments conducted in Ecole Centrale in Paris during his trial, were assisted buy several eminent French scientist. But although the judge sentenced, that Dunikowki's "secret process for turning sand into gold is an impracticable combination of absurdities and contradictions," Polish engineer was still able to find other backers after being released from French prison. We find the traces of his further activity in Italy, Switzerland, Belgium and Philippines. Finally, in early 1950s he ended his journey in the United States as a political refugee.


Excellent prognosis in a subset of patients with Ewing sarcoma identified at diagnosis by CD56 using flow cytometry.
Ewing sarcoma (ES) is considered a systemic disease with the majority of patients harboring micrometastases at diagnosis. Multiparameter flow cytometry (MPFC) was used to detect ES cells in bone marrow (BM) of ES patients at diagnosis and to evaluate the prognostic significance of CD56 expression in BM samples. BM samples from 46 ES patients, 6 tumor aspirates, 2 ES cell lines, and 10 control BM samples were analyzed by MPFC. ES cells were identified by the combination of CD45-/CD90+/CD99+. CD56 was evaluated on these cells by a cutoff of 22%. BM samples obtained from all patients at diagnosis were found to be positive for micrometastatic tumor cells assessed by CD99+/CD90+/CD45- expression. A total of 60% of the BM samples harbored high CD56 expression. There was a highly significant correlation between CD56 expression and progression-free survival (PFS; 69% in low/negative expression versus 30% in high expression groups, P = 0.024). In patients with localized nonpelvic disease, those expressing low/negative CD56 had 100% PFS versus 40% in the high expressing group (P = 0.02). By Cox regression analysis, CD56 was found to be an independent prognostic marker with an 11-fold increased risk for relapse in patients with localized disease (P = 0.006). All samples contained cells that are positive for the CD99+/CD90+/CD45- combination at diagnosis, indicating that ES is a systemic disease. CD56 expression could be used to reveal ES patients with excellent prognosis or patients predisposed to relapse, thus improving treatment stratification and implementation of personalized therapy.


Ultimobranchial body cyst in an adult horse: clinical, histopathologic, and immunohistochemical features.
A 9-y-old Mangalarga Marchador gelding was referred to a veterinary hospital because of a swelling on the upper right side of the neck. Ultrasound examination revealed a multilocular structure adjacent to the thyroid gland with low echogenic content suggestive of fluid. The mass was removed surgically. Histologically, the cystic cavities in the surgical sample were filled with abundant eosinophilic secreta and lined by cuboidal, segmentally ciliated, columnar epithelium with interspersed goblet cells. Segmental crowding of the multilayered lining of the cyst was noted. Immunohistochemistry suggested the presence of both C cells and follicular cells, given the positivity of the immunomarkers calcitonin and TTF-1, respectively. The histogenesis of ultimobranchial cysts is uncertain. Based on clinical, histopathologic, and immunohistochemical identification, the cystic structure in this case is compatible with an ultimobranchial body cyst.


Universal scaling of local plasmons in chains of metal spheres.
The position, width, extinction, and electric field of localized plasmon modes in closely-coupled linear chains of small spheres are investigated. A dipole-like model is presented that separates the universal geometric factors from the specific metal permittivity. An electrostatic surface integral method is used to deduce universal parameters that are confirmed against results for different metals (bulk experimental Ag, Au, Al, K) calculated using retarded vector spherical harmonics and finite elements. The mode permittivity change decays to an asymptote with the number of particles in the chain, and changes dramatically from 1/f(3) to 1/f(1/2) as the gap fraction (ratio of gap between spheres to their diameter), f, gets smaller. Scattering increases significantly with closer coupling. The mode sharpness, strength and electric field for weakly retarded calculations are consistent with electrostatic predictions once the effect of radiative damping is accounted for.


Molecular cloning of goat 20alpha-hydroxysteroid dehydrogenase cDNA.
20Alpha-hydroxysteroid dehydrogenase (20alpha-HSD), which catalyzes the conversion of progesterone to its inactive form 20alpha-dihydroprogesterone, is expressed in murine placenta and has been suggested to play roles in maintaining pregnancy. To understand the role of 20alpha-HSD during pregnancy in the goat, as a first step, cloning and sequencing of 20alpha-HSD cDNA were performed. The full nucleotide sequence of 20alpha-HSD cDNA was determined on samples obtained from the corpus luteum at the luteal phase of the estrous cycle and the placenta in late pregnancy by RT-PCR and 3' and 5' RACE systems. Cloned 20alpha-HSD cDNA consisted of 1124 bp and belonged to the aldo-keto reductase superfamily. From the start codon to stop codon there were 323 amino acids, the same as in other species. To verify whether the protein derived from goat 20alpha-HSD cDNA had 20alpha-HSD activity, the cDNA was expressed by bacteria. Bacterially expressed goat 20alpha-HSD protein showed 20alpha-HSD enzyme activity. A tissue distribution study demonstrated that 20alpha-HSD was expressed in the placenta, but not in the adrenal gland, liver and spleen during pregnancy. The present study suggests that goat 20alpha-HSD is another member of the aldo-keto reductase superfamily and that it plays a role in the placenta during pregnancy.


Optimization of caseinate-coated simvastatin-zein nanoparticles: improved bioavailability and modified release characteristics.
The current study focuses on utilization of the natural biocompatible polymer zein to formulate simvastatin (SMV) nanoparticles coated with caseinate, to improve solubility and hence bioavailability, and in addition, to modify SMV-release characteristics. This formulation can be utilized for oral or possible depot parenteral applications. Fifteen formulations were prepared by liquid-liquid phase separation method, according to the Box-Behnken design, to optimize formulation variables. Sodium caseinate was used as an electrosteric stabilizer. The factors studied were: percentage of SMV in the SMV-zein mixture (X1), ethanol concentration (X2), and caseinate concentration (X3). The selected dependent variables were mean particle size (Y1), SMV encapsulation efficiency (Y2), and cumulative percentage of drug permeated after 1 hour (Y3). The diffusion of SMV from the prepared nanoparticles specified by the design was carried out using an automated Franz diffusion cell apparatus. The optimized SMV-zein formula was investigated for in vivo pharmacokinetic parameters compared with an oral SMV suspension. The optimized nanosized SMV-zein formula showed a 131 nm mean particle size and 89% encapsulation efficiency. In vitro permeation studies displayed delayed permeation characteristics, with about 42% and 85% of SMV cumulative amount released after 12 and 48 hours, respectively. Bioavailability estimation in rats revealed an augmentation in SMV bioavailability from the optimized SMV-zein formulation, by fourfold relative to SMV suspension. Formulation of caseinate-coated SMV-zein nanoparticles improves the pharmacokinetic profile and bioavailability of SMV. Accordingly, improved hypolipidemic activities for longer duration could be achieved. In addition, the reduced dosage rate of SMV-zein nanoparticles improves patient tolerability and compliance.


Anaemia, renal dysfunction and in-hospital outcomes in patients with heart failure in Botswana.
Anaemia and renal dysfunction are associated with an increased morbidity and mortality in heart failure (HF) patients. To estimate the frequency and impact of anaemia and renal dysfunction on in-hospital outcomes in patients with HF. A total of 193 consecutive patients with HF admitted to Princess Marina Hospital, Gaborone, Botswana, from February 2014 to February 2015, were studied. Anaemia was defined as haemoglobin <13 g/dL for men and <12 g/dL for women. Renal dysfunction was defined by an estimated glomerular filtration rate (eGFR) <60 mL/min/1.73 m2, calculated by the simplified Modification of Diet in Renal Disease formula. The in-hospital outcomes included length of hospital stay and mortality. The mean (standard deviation (SD)) age was 54.2 (17.1) years and 53.9% of the patients were men. The overall median eGFR was 75.9 mL/min/1.73 m2 and renal dysfunction was detected in 60 (31.1%) patients. Renal dysfunction was associated with hypertension (p=0.01), diabetes mellitus (p=0.01) and a lower haemoglobin level (p=0.008). The mean (SD) haemoglobin was 12.0 (3.0) g/dL and 54.9% of the patients were anaemic. Microcytic, normocytic and macrocytic anaemia were found in 32.1%, 57.5% and 10.4% of patients, respectively. The mean (SD) haemoglobin level for males was significantly higher than for females (12.4 (3.3) g/dL v. 11.5 (2.5) g/dL; p=0.038). Anaemia was more common in patients with diabetes (p=0.028) and in those with increased left ventricular ejection fraction (p=0.005). Neither renal dysfunction nor anaemia was significantly associated with the length of hospital stay or in-hospital mortality. Anaemia and renal dysfunction are prevalent in HF patients, but neither was an independent predictor of length of stay or in-hospital mortality in this population. These findings indicate that HF data in developed countries may not apply to countries in sub-Saharan Africa, and call for more studies to be done in this region.


[A psychosocial perspective of depressive symptoms in adolescence].
This study aimed at understanding the social representations (SR) of depression among adolescents on high school level. Two hundred and twenty-two (222) adolescents of both sexes (53% male and 47% female), aged between 14 and 19 years and attending the first, second and third grades of high school participated in the study. The instruments used were: the Beck Depression Inventory (BDI) for selecting the sample, using 17 as a cut point. The adolescents who achieved this score were invited to participate in the second part of this study consisting of semi-structured interviews and application of Free Association Tests with the stimulus words: depression, depressed person and me. It was verified that the adolescents anchored their SR of depression as synonym for pain, grief, misfortune, hatred, death, discouragement, anxiety, solitude and tears. The SR elaborated by the social actors of this study revealed a similarity with the conception/description, as described in psychiatry. The purpose of this study besides helping to understand the suffering provoked by depressive symptoms is to contribute to a better quality of life of these adolescents.


UHRF1 suppresses retrotransposons and cooperates with PRMT5 and PIWI proteins in male germ cells.
DNA methylation, repressive histone marks, and PIWI-interacting RNA (piRNA) are essential for the control of retrotransposon silencing in the mammalian germline. However, it remains unknown how these repressive epigenetic pathways crosstalk to ensure retrotransposon silencing in the male germline. Here, we show that UHRF1 is responsible for retrotransposon silencing and cooperates with repressive epigenetic pathways in male germ cells. Conditional loss of UHRF1 in postnatal germ cells causes DNA hypomethylation, upregulation of retrotransposons, the activation of a DNA damage response, and switches in the global chromatin status, leading to complete male sterility. Furthermore, we show that UHRF1 interacts with PRMT5, an arginine methyltransferase, to regulate the repressive histone arginine modifications (H4R3me2s and H3R2me2s), and cooperates with the PIWI pathway during spermatogenesis. Collectively, UHRF1 regulates retrotransposon silencing in male germ cells and provides a molecular link between DNA methylation, histone modification, and the PIWI pathway in the germline.


[Limits of optimizing resource allocation by debiting the public health service].
Based on the Dornier consulting opinion on the public health service system of Berlin, the limits of optimising the resource allocation process are discussed with respect to austerity budgets. The discussion focuses firstly on problems of finding a consensus of opinion about public tasks and purposes in the German settings of pluralistic impact on health policy, and secondly on methodical, functional and practical limits. Methodical arguments are supported by a comparison of personal density with regard to the twelve largest cities of Western Germany, and supplemented by a factor analysis of sociodemographic characteristics in order to quantify the need of social work in an urban context. Functional and practical limits are demonstrated with respect to the realisation of the principle of subsidiary and concerning the practice of wholesale cut-downs of public health services budget.


Proteinase-activated receptor-2-activating peptides induce leukocyte rolling, adhesion, and extravasation in vivo.
Proteinase-activated receptor 2 (PAR2) has been suggested to play a role in inflammatory reactions. Because leukocyte-endothelial cell interactions are critical events during inflammatory reactions, and because PAR2 is expressed both on endothelium and leukocytes, we have examined the effects of PAR2-activating peptides (PAR2-APs) on leukocyte rolling and adhesion in mesenteric venules and on leukocyte recruitment into the peritoneal cavity. Using intravital microscopy, leukocyte rolling, flux, and adhesion in rat mesenteric postcapillary venules were quantified. Topical addition of PAR2-APs (10 microM) for 1 min to the superfused venule induced a significant increase in leukocyte rolling and adherence. The increase in leukocyte adherence was not affected by pretreatment with a mast cell stabilizer (sodium cromoglycate) nor by prior degranulation of mast cells with compound 48/80. Nonetheless, both leukocyte rolling and adhesion were completely inhibited by pretreatment with a platelet-activating factor receptor antagonist (WEB 2086). Intraperitoneal injections of a selective PAR2-AP (SLIGRL-NH2) caused a significant increase in leukocyte migration into the peritoneal cavity. The effect of SLIGRL-NH2 on peritoneal leukocyte infiltration was completely inhibited by WEB 2086. These data suggest that PAR2 activation could contribute to several early events in the inflammatory reaction, including leukocyte rolling, adherence, and recruitment, by a mechanism dependent on platelet-activating factor release.


Nebulizer use in inner-city children with asthma: morbidity, medication use, and asthma management practices.
To assess the frequency of nebulizer use, describe morbidity and patterns of medication administration, and examine the potential relationships between inhaled anti-inflammatory medication administration, asthma morbidity, and asthma management practices in children with asthma using a nebulizer compared with children with asthma not using a nebulizer. A cross-sectional, descriptive survey of previous events. Elementary schools and participants' homes in Baltimore, Md, and Washington, DC. Six hundred eighty-six families of children aged 5 to 12 years with a diagnosis of at least mild, persistent asthma. None. Asthma morbidity, health care utilization, pattern of asthma medication administration, nebulizer use, and asthma management data were collected by telephone survey administered to caregivers. Nebulizer use was defined as use at least 1 or more days per month during the last 6 months. Of 686 children identified, 231 (33%) reported current nebulizer use. Nebulizer users had significantly increased lifetime hospital admissions, hospitalizations, and emergency department visits in the last 6 months compared with nonnebulizer users. Inhaled corticosteroid administration was low for both groups (nonnebulizer users, 8%; nebulizer users, 15%). In the nebulizer users group, administration of inhaled anti-inflammatory medications was associated with increased asthma morbidity (increased hospitalizations, days and nights with symptoms, and oral steroid use). Nebulizer use by inner-city children with asthma is higher than anticipated but is not associated with reduced asthma morbidity. This group of high-risk children was undertreated with inhaled corticosteroids for long-term control of asthma despite reports of adequate monitoring by a primary care physician.


Ranibizumab in retinal angiomatous proliferation (RAP): influence of RAP stage on visual outcome.
To evaluate the influence of retinal angiomatous proliferation (RAP) stage on visual and anatomic outcome after ranibizumab (Lucentis®). This was a prospective study on consecutively diagnosed RAP eyes at the Hospital Clínico San Carlos, Madrid. Best-corrected visual acuity (BCVA) and optical coherence tomography (OCT) are performed monthly. Indocyanine green angiography (ICG) and fluorescein angiography (FA) are performed at baseline and every 3 months thereafter. A starting dose of a monthly ranibizumab injection in the first 3 months is followed by retreatment in case of intraretinal edema, subretinal fluid, or pigment epithelium detachment (PED) in OCT, increased leakage in FA, or a hot spot in ICG. A total of 53 eyes from 49 patients were included. The mean change in BCVA at 12 months was +7.3, +0.83, and -2.1 letters in stages IIA (21 cases), II B (18 cases), and III (14 cases), respectively. After adjusting the change in BCVA according to baseline BCVA, ß coefficient was -6.012 letters (p=0.025) in stage IIB and -9.762 letters (p=0.003) in stage III vs stage IIA. Four cases had a retinal pigment epithelium tear after injection of ranibizumab. Patients in stage II without PED have a better visual and anatomic evolution than patients in stage II with PED and stage III.


Leadership behavior and subordinate well-being.
The authors used a longitudinal design to investigate the relation between leadership behavior and the well-being of subordinates. Well-being is conceptualized as people's feelings about themselves and the settings in which they live and work. Staff members (N = 562) of 2 Community Trusts participated 4 times in a 14-month period. Five models were formulated to answer 2 questions: What is the most likely direction of the relation between leadership and well-being, and what is the time frame of this relation? The model with the best fit suggested that leadership behavior and subordinate responses are linked in a feedback loop. Leadership behavior at Time 1 influenced leadership behavior at Time 4. Subordinate well-being at Time 2 synchronously influenced leadership behavior at Time 2. Leadership behavior at Time 4 synchronously influenced subordinate well-being at Time 4.


Nafion-Mediated Proton Transfer Facilitates the Electroreduction of SO2 to H2S.
The electrocatalytic reduction of SO2 to produce H2S is a critical approach for achieving the efficient utilization of sulfur resources. At the core of this approach for commercial applications lies the imperative need to elevate current density. However, the challenges posed by high current density manifest in the rapid depletion of protons, leading to a decrease in SO2 partial pressure, consequently hampering the generation and separation of H2S. Here, we demonstrate an effective solution to alleviate the problem of insufficient supply of protons by employing Nafion polymer as the proton conductor to modified Cu catalysts surface, creating a proton-enriched layer to boost H2S generation. It was observed that Nafion shortens the hydrogen bonds with water molecules in the electrolyte via its sulfonic acid groups, benefiting the proton transfer and consequently increasing the proton density on the electrode surface by 5-fold. With the Nafion-modified catalyst, the H2S partial current density and separation efficiency reached 205.9 mA·cm-2 (1.01 mmol·cm-2·h-1) and 87.8%, which were 1.34 and 1.22 times that on unmodified Cu, respectively. This work highlights the practicality of fabricating a proton conductor via ionic polymer for the control over product selectivity in pH-sensitive reactions under high current density.


[Determining the levels of staphylococcal enterotoxins types A and B in food products by the immunoenzyme method].
"Sandwich" variant of ELISA was used to identify staphylococcal enterotoxins (SE), types A and B, in S. aureus filtrates inducing food poisoning, in extracts of the lactic acid product for infants "Biphilin" that caused staphylococcal infection, and in foods contaminated with SE in varying concentrations. It has been shown that ELISA used for SE identification in foods permits revealing SE, types A and B, in liquid products in concentrations of 1-2 ng/ml (that is 1000-fold more sensitive, than the immunodiffusion test, 400-800-fold more sensitive than the passive hemagglutination test, and 10-fold more sensitive than the indirect passive hemagglutination test), and in solid products--in concentrations of 5-10 ng/g (after artificial contamination).


COVID-19, Influenza and RSV: Surveillance-informed prevention and treatment - Meeting report from an isirv-WHO virtual conference.
The International Society for Influenza and other Respiratory Virus Diseases (isirv) and the WHO held a joint virtual conference from 19th-21st October 2021. While there was a major focus on the global response to the SARS-CoV-2 pandemic, including antivirals, vaccines and surveillance strategies, papers were also presented on treatment and prevention of influenza and respiratory syncytial virus (RSV). Potential therapeutics for SARS-CoV-2 included host-targeted therapies baricitinib, a JAK inhibitor, tocilizumab, an IL-6R inhibitor, verdinexor and direct acting antivirals ensovibep, S-217622, AT-527, and monoclonal antibodies casirivimab and imdevimab, directed against the spike protein. Data from trials of nirsevimab, a monoclonal antibody with a prolonged half-life which binds to the RSV F-protein, and an Ad26.RSV pre-F vaccine were also presented. The expanded role of the WHO Global Influenza Surveillance and Response System to address the SARS-CoV-2 pandemic was also discussed. This report summarizes the oral presentations given at this meeting for the benefit of the broader medical and scientific community involved in surveillance, treatment and prevention of respiratory virus diseases.


Are Vancomycin Trough Concentrations of 15 to 20 mg/L Associated With Increased Attainment of an AUC/MIC ≥ 400 in Patients With Presumed MRSA Infection?
To determine whether there is an association between higher vancomycin trough concentrations and attainment of a calculated area under the concentration-time curve (AUC)/minimum inhibitory concentration (MIC) ≥400. A retrospective analysis was conducted among vancomycin-treated adult patients with a positive methicillin-resistant Staphylococcus aureus (MRSA) culture. Attainment of a calculated AUC/MIC ≥400 was compared between patients with troughs in the reference range of 15 to 20 mg/L and those with troughs in the following ranges: <10, 10 to 14.9, and >20 mg/L. Nephrotoxicity was assessed as a secondary outcome based on corrected average vancomycin troughs over 10 days of treatment. Overall, 226 patients were reviewed and 100 included. Relative to troughs ≥10, patients with vancomycin troughs <10 mg/L were 73% less likely to attain an AUC/MIC ≥400 (odds ratio [OR] 0.27, 95% confidence interval [CI]: 0.01-0.75). No difference was found in the attainment of an AUC/MIC ≥400 in patients with troughs of 10 to 14.9 mg/L and >20 mg/L when compared to patients with troughs of 15 to 20 mg/L. The mean corrected average vancomycin trough was higher in patients developing nephrotoxicity compared to those who did not (19.5 vs 14.5 mg/L, P < .001). Achieving vancomycin serum trough concentrations of 15 to 20 mg/L did not result in an increased attainment of the AUC/MIC target relative to troughs of 10 to 14.9 mg/L but may increase nephrotoxicity risk.


Single-center retrospective review of ultrasound-accelerated versus traditional catheter-directed thrombolysis for acute lower extremity deep venous thrombosis.
Objective Systemic anticoagulation remains the standard for acute lower extremity (LE) deep venous thrombosis (DVT), but growing interest in catheter-directed thrombolysis (CDT) and its potential to reduce the incidence of post-thrombotic syndrome (PTS) has led to advent of ultrasound-accelerated CDT (US-CDT). Few studies to date have examined the outcomes of US-CDT against traditional CDT (T-CDT). Methods This is a retrospective, single-center review of all patients treated for acute LE DVT over a five-year period with either US- and T-CDT. Patients were stratified based on demographics, presentation, co-morbidities, risk factors, and peri-procedural data. Results Seventy-six limbs in 67 patients were treated; 51 limbs in 42 patients were treated with US-CDT, and 25 limbs in 25 patients were treated with T-CDT. Adjuncts include: pharmacomechanical thrombolysis ( n = 28 vs. 20, p = 0.04), angioplasty ( n = 22 vs. 18, p = 0.11), stenting ( n = 30 vs. 6, p ≤ 0.001), and IVC filter insertion ( n = 5 vs. 0, p = 0.07). Mean lysis times were 21 ± 1.7 and 24 ± 1.8 h for US- and T-CDT, respectively ( p = 0.26). Thirty (25 ultrasound, 5 traditional) limbs had complete lysis. Thirty-one (22 ultrasound, 9 traditional) limbs had incomplete lysis. Fifteen (4 ultrasound, 11 traditional) limbs had ineffective lysis ( p = 0.002 in favor of ultrasound). Four patients (3 US-CDT, 1 T-CDT) had recurrent ipsilateral thrombosis within 30 days ( p = 0.60). By Kaplan-Meier analysis, there were no significant difference between primary patency, primary-assisted patency, secondary patency, re-thrombosis, and recurrent symptoms at 6, 12, and 24 months. Conclusion US-CDT does not significantly improve mid-term patencies but results in greater acute clot burden reduction in patients with acute LE DVTs compared to T-CDT, which may be beneficial in reducing the long-term incidence of PTS.


The effect of electrical stimulation therapies on spinal fusion: a cross-disciplinary systematic review and meta-analysis of the preclinical and clinical data.
Nonunion is a common complication of spinal fusion surgeries. Electrical stimulation technologies (ESTs)-namely, direct current stimulation (DCS), capacitive coupling stimulation (CCS), and inductive coupling stimulation (ICS)-have been suggested to improve fusion rates. However, the evidence to support their use is based solely on small trials. Here, the authors report the results of meta-analyses of the preclinical and clinical data from the literature to provide estimates of the overall effect of these therapies at large and in subgroups. A systematic review of the English-language literature was performed using PubMed, Embase, and Web of Science databases. The query of these databases was designed to include all preclinical and clinical studies examining ESTs for spinal fusion. The primary endpoint was the fusion rate at the last follow-up. Meta-analyses were performed using a Freeman-Tukey double arcsine transformation followed by random-effects modeling. A total of 33 articles (17 preclinical, 16 clinical) were identified, of which 11 preclinical studies (257 animals) and 13 clinical studies (2144 patients) were included in the meta-analysis. Among preclinical studies, the mean fusion rates were higher among EST-treated animals (OR 4.79, p < 0.001). Clinical studies similarly showed ESTs to increase fusion rates (OR 2.26, p < 0.001). Of EST modalities, only DCS improved fusion rates in both preclinical (OR 5.64, p < 0.001) and clinical (OR 2.13, p = 0.03) populations; ICS improved fusion in clinical studies only (OR 2.45, p = 0.014). CCS was not effective at increasing fusion, although only one clinical study was identified. A subanalysis of the clinical studies found that ESTs increased fusion rates in the following populations: patients with difficult-to-fuse spines, those who smoke, and those who underwent multilevel fusions. The authors found that electrical stimulation devices may produce clinically significant increases in arthrodesis rates among patients undergoing spinal fusion. They also found that the pro-arthrodesis effects seen in preclinical studies are also found in clinical populations, suggesting that findings in animal studies are translatable. Additional research is needed to analyze the cost-effectiveness of these devices.


Free fatty acid palmitate impairs the vitality and function of cultured human bladder smooth muscle cells.
Incidence of urinary tract infections is elevated in patients with diabetes mellitus. Those patients show increased levels of the saturated free fatty acid palmitate. As recently shown metabolic alterations induced by palmitate include production and secretion of the pro-inflammatory cytokine interleukine-6 (IL-6) in cultured human bladder smooth muscle cells (hBSMC). Here we studied the influence of palmitate on vital cell properties, for example, regulation of cell proliferation, mitochondrial enzyme activity and antioxidant capacity in hBSMC, and analyzed the involvement of major cytokine signaling pathways. HBSMC cultures were set up from bladder tissue of patients undergoing cystectomy and stimulated with palmitate. We analyzed cell proliferation, mitochondrial enzyme activity, and antioxidant capacity by ELISA and confocal immunofluorescence. In signal transduction inhibition experiments we evaluated the involvement of NF-κB, JAK/STAT, MEK1, PI3K, and JNK in major cytokine signaling pathway regulation. We found: (i) palmitate decreased cell proliferation, increased mitochondrial enzyme activity and antioxidant capacity; (ii) direct inhibition of cytokine receptor by AG490 even more strongly suppressed cell proliferation in palmitate-stimulated cells, while counteracting palmitate-induced increase of antioxidant capacity; (iii) in contrast knockdown of the STAT3 inhibitor SOCS3 increased cell proliferation and antioxidant capacity; (iv) further downstream JAK/STAT3 signaling cascade the inhibition of PI3K or JNK enhanced palmitate induced suppression of cell proliferation; (v) increase of mitochondrial enzyme activity by palmitate was enhanced by inhibition of PI3K but counteracted by inhibition of MEK1. Saturated free fatty acids (e.g., palmitate) cause massive alterations in vital cell functions of cultured hBSMC involving distinct major cytokine signaling pathways. Thereby, certain cytokines might counteract the palmitate-induced downregulation of cell proliferation and vitality. This could be an important link to clinical findings of increased risk of metabolic related bladder diseases such as overactive bladder (OAB) and bladder pain syndrome/interstitial cystitis (BPS/IC).


Effect of cysteine residues on the activity of arginyl-tRNA synthetase from Escherichia coli.
Arginyl-tRNA synthetase (ArgRS) from Escherichia coli (E. coli) contains four cysteine residues. In this study, the role of cysteine residues in the enzyme has been investigated by chemical modification and site-directed mutagenesis. Titration of sulfhydryl groups in ArgRS by 5, 5'-dithiobis(2-nitro benzoic acid) (DTNB) suggested that a disulfide bond was not formed in the enzyme and that, in the native condition, two DTNB-sensitive cysteine residues were located on the surface of ArgRS, while the other two were buried inside. Chemical modification of the native enzyme by iodoacetamide (IAA) affected only one DTNB-sensitive cysteine residue and resulted in 50% loss of enzyme activity, while modification by N-ethylmeimide (NEM) affected two DTNB-sensitive residues and caused a complete loss of activity. These results, when combined with substrate protection experiments, suggested that at least the two cysteine residues located on the surface of the molecule were directly involved in substrates binding and catalysis. However, changing Cys to Ala only resulted in slight loss of enzymatic activity and substrate binding, suggesting that these four cysteine residues in E. coli ArgRS were not essential to the enzymatic activity. Moreover, modifications of the mutant enzymes indicated that the two DTNB- and NEM-sensitive residues were Cys(320) and Cys(537) and the IAA-sensitive was Cys(320). Our study suggested that inactivation of E. coli ArgRS by sulfhydryl reagents is a result of steric hindrance in the enzyme.


Quality control of laboratory methods for semen evaluation in a multicenter research study.
Rigorously standardized laboratory protocols and strict quality control (QC) are essential for meaningful comparisons between semen quality data from multiple sites. We describe our experience with the Study for Future Families (SFF), a multicenter study of semen quality in the United States. Detailed protocols were developed, and technicians from each study site attended a training session at the central laboratory. Technicians received blinded replicates from diluted semen specimens for counting by MicroCell and hemacytometer. Sperm motility was assessed using videotaped recordings for simple percent motility and categorical assessment of individual sperm progression as recommended by the World Health Organization (WHO). The mean intertechnician coefficient of variation for individual specimens was 12.6% for MicroCell counts, 15.2% for hemacytometer counts, and 10.5% for percent motility. Intratechnician coefficients of variation averaged 10.3% for MicroCell counts, 12.5% for hemacytometer counts, and 5.2% for percent motility. The average percent differences between the technicians' values and the central standard for individual specimens were 13.5%, 16.6%, and 11.9% for MicroCell counts, hemacytometer counts, and simple percent motility, respectively. We achieved our goal of maintaining mean intratechnician coefficients of variation and mean percent differences from the standard values of 15% or less for measurements of simple percent motility and sperm concentration by MicroCell. Standardization using the Improved Neubauer hemacytometer chamber proved more difficult. We were not successful in standardizing a method for categorical assessment of individual sperm progression.


Tumor-associated myeloid cells: new understandings on their metabolic regulation and their influence in cancer immunotherapy.
Tumor-associated myeloid cells (TAMCs), mainly represented by tumor-associated macrophages and myeloid-derived suppressor cells, can promote tumor growth directly, by favoring tumor cell proliferation and survival, and indirectly, by creating an immunosuppressive microenvironment. Myeloid cells are characterized by an extreme phenotypical and functional plasticity. Immunometabolism is now emerging as a crucial aspect of TAMCs skewing toward pro-tumoral activities. The metabolic re-education of myeloid cells is a new strategy to boost their antitumor effector functions. Several anticancer therapies targeting TAMCs are already under investigation. Nowadays, the hot topic of cancer immunotherapy is represented by immune checkpoint inhibitors. These drugs unrestrain T-cell-mediated tumor elimination by removing suppressive signals delivered by tumor-associated cells. The efficacy of immune checkpoint blockade can be enhanced using coordinated strategies to counteract the TAMCs-dependent impairment of immune adaptive responses. In the first part of the review, we will describe the association between metabolic reprogramming and TAMCs biological activities. In the second part, we will illustrate the potential of combination therapies associating TAMC-targeting drugs with immune checkpoint inhibitors.


Chinese Newlyweds' Perception and Tolerance Toward Common and Severe Partner Aggression.
This study aimed to find the judgment demarcation points of Chinese newlyweds toward common and severe partner aggression, exploring the correlation of asymmetrical commitment and partner aggression tolerance, and revealing the gender differences in aggression tolerance. We conducted two online questionnaire surveys with a total of 629 Chinese newlyweds. Specifically, data for group 1 were collected from 326 Chinese newlyweds for exploratory factor analysis of aggression normality, and data for group 2 from the remaining 303 couples were used for confirmatory factor analysis and inferential statistical analyses. Results showed that eight items representing non-physical aggression were regarded as common aggression, seven items indicating physical aggression were regarded as severe aggression, and one item was deleted because of disqualification in the exploratory factor analysis. Moreover, individuals showed greater tolerance toward common aggression compared with severe aggression. In terms of commitment, the 303 couples were divided into two groups: asymmetrically committed relationships (ACR) and non-asymmetrically committed relationships (non-ACRs). Through multilevel modeling, we found that couples in ACRs had a greater tolerance for common aggression. In addition, tolerance showed gender differences: husbands displayed a more tolerant attitude toward partner aggression, whether common or severe types. The study found the demarcation points of aggression normality in Chinese newlyweds broadened the application of commitment in research on partner aggression and emphasized the importance of study of dyadic relationships.


Nanoparticlized System: Promising Approach for the Management of Alzheimer's Disease through Intranasal Delivery.
Alzheimer's disease (AD) is a neurodegenerative brain problem and responsible for causing dementia in aged people. AD has become most common neurological disease in the elderly population worldwide and its treatment remains still challengeable. Therefore, there is a need of an efficient drug delivery system which can deliver the drug to the target site. Nasal drug delivery has been used since prehistoric times for the treatment of neurological disorders like Alzheimer's disease (AD). For delivering drug to the brain, blood brain barrier (BBB) is a major rate limiting factor for the drugs. The desired drug concentration could not be achieved through the conventional drug delivery system. Thus, nanocarrier based drug delivery systems are promising for delivering drug to brain. Nasal route is a most convenient for targeting drug to the brain. Several factors and mechanisms need to be considered for an effective delivery of drug to the brain particularly AD. Various nanoparticlized systems such as nanoparticles, liposomes, exosomes, phytosomes, nanoemulsion, nanosphere, etc. have been recognized as an effective drug delivery system for the management of AD. These nanocarriers have been proven with improved permeability as well as bioavailability of the anti-Alzheimer's drugs. Some novel drug delivery systems of anti-Alzheimer drugs are under investigation of different phase of clinical trials. Present article highlights on the nanotechnology based intranasal drug delivery system for the treatment of Alzheimer's disease. Furthermore, consequences of AD, transportation mechanism, clinical updates and recent patents on nose to brain delivery for AD have been discussed.


Evaluation of the most commonly used (semi-)quantitative parameters of 18F-FDG PET/CT to detect malignant transformation of neurofibromas in neurofibromatosis type 1.
In patients with neurofibromatosis type 1, transformation of neurofibromas into a malignant peripheral nerve sheath tumor (MPNST) is a severe complication of the disease. Fluorine-18-fluorodeoxyglucose PET/computed tomography (PET/CT) is a viable option for detecting malignant tumors in neurofibromatosis type 1 patients. The aim of this review was to assess the diagnostic performance of the most frequently used parameters of PET/CT in detecting MPNST. An extensive computer search was performed using the Cochrane Library, Pubmed, and Medline/Embase databases. Two reviewers independently extracted data of relevant studies and assessed the methodological quality (QUADAS-2). The diagnostic performance of PET/CT parameters in individual studies was determined by calculating a diagnostic odds ratio (DOR) using the absolute numbers of true-positive, true-negative, false-positive, and false-negative test results. A total of eight studies were included, of which three evaluated the standardized uptake value as a diagnostic parameter, two assessed the tumor-to-liver (T/L) ratio, and three articles described both parameters. The cut-off values for maximum standardized uptake value (SUVmax) ranged from 3.2 to 4.5; for the T/L ratio, the cut-off values were between 1.0 and 4.3. The sensitivity and specificity ranged from 90 to 100% and from 80 to 100%, respectively (SUVmax). T/L ratios were associated with 92-100% sensitivity and 72-94% specificity. The corresponding DORs ranged from 57 to 145 (SUVmax) and 35 to 655 (T/L ratio). Both the SUV and the T/L ratio are associated with high sensitivity combined with acceptable specificity in detecting MPNST. There is a tendency toward higher DORs using the T/L ratio, but the number of studies is limited.


A stripe rust effector Pst18363 targets and stabilises TaNUDX23 that promotes stripe rust disease.
Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), poses a tremendous threat to the production of wheat worldwide. The molecular mechanisms of Pst effectors that regulate wheat immunity are poorly understood. In this study, we identified an effector Pst18363 from Pst that suppresses plant cell death in Nicotiana benthamiana and in wheat. Knocking down Pst18363 expression by virus-mediated host-induced gene silencing significantly decreased the number of rust pustules, indicating that Pst18363 functions as an important pathogenicity factor in Pst. Pst18363 was proven to interact with wheat Nudix hydrolase 23 TaNUDX23. In wheat, silencing of TaNUDX23 by virus-induced gene silencing increased reactive oxygen species (ROS) accumulation induced by the avirulent Pst race CYR23, whereas overexpression of TaNUDX23 suppressed ROS accumulation induced by flg22 in Arabidopsis. In addition, TaNUDX23 suppressed Pst candidate effector Pst322-trigged cell death by decreasing ROS accumulation in N. benthamiana. Knocking down of TaNUDX23 expression attenuated Pst infection, indicating that TaNUDX23 is a negative regulator of defence. In N. benthamiana, Pst18363 stabilises TaNUDX23. Overall, our data suggest that Pst18363 stabilises TaNUDX23, which suppresses ROS accumulation to facilitate Pst infection.


A crisis emerges: Lesbian health between breast cancer and HIV/AIDS.
In this essay, I argue that lesbians have come to be a population of concern for state-based health organizations as a result of lesbian health activism that drew connections between breast cancer and HIV/AIDS. In order to develop this analysis, I tell the story of the rise of lesbian breast cancer activism in concert with HIV/AIDS in San Francisco in the early 1990s. The state recognition of lesbian health needs, and with it the solidification of lesbian as a biopolitical category, was catalyzed by associations with the AIDS crisis and HIV activism, but also required an articulated difference, or lesbian specificity, which breast cancer provided. And yet, documenting the multiple, and potentially contradictory, ways that these associations were made resists understanding "lesbian" as a static category.


Variation in lithic technological strategies among the Neanderthals of Gibraltar.
THE EVIDENCE FOR NEANDERTHAL LITHIC TECHNOLOGY IS REVIEWED AND SUMMARIZED FOR FOUR CAVES ON THE ROCK OF GIBRALTAR: Vanguard, Beefsteak, Ibex and Gorham's. Some of the observed patterns in technology are statistically tested including raw material selection, platform preparation, and the use of formal and expedient technological schemas. The main parameters of technological variation are examined through detailed analysis of the Gibraltar cores and comparison with samples from the classic Mousterian sites of Le Moustier and Tabun C. The Gibraltar Mousterian, including the youngest assemblage from Layer IV of Gorham's Cave, spans the typical Middle Palaeolithic range of variation from radial Levallois to unidirectional and multi-platform flaking schemas, with characteristic emphasis on the former. A diachronic pattern of change in the Gorham's Cave sequence is documented, with the younger assemblages utilising more localized raw material and less formal flaking procedures. We attribute this change to a reduction in residential mobility as the climate deteriorated during Marine Isotope Stage 3 and the Neanderthal population contracted into a refugium.


Blood pressure and incidence of twelve cardiovascular diseases: lifetime risks, healthy life-years lost, and age-specific associations in 1·25 million people.
The associations of blood pressure with the different manifestations of incident cardiovascular disease in a contemporary population have not been compared. In this study, we aimed to analyse the associations of blood pressure with 12 different presentations of cardiovascular disease. We used linked electronic health records from 1997 to 2010 in the CALIBER (CArdiovascular research using LInked Bespoke studies and Electronic health Records) programme to assemble a cohort of 1·25 million patients, 30 years of age or older and initially free from cardiovascular disease, a fifth of whom received blood pressure-lowering treatments. We studied the heterogeneity in the age-specific associations of clinically measured blood pressure with 12 acute and chronic cardiovascular diseases, and estimated the lifetime risks (up to 95 years of age) and cardiovascular disease-free life-years lost adjusted for other risk factors at index ages 30, 60, and 80 years. This study is registered at ClinicalTrials.gov, number NCT01164371. During 5·2 years median follow-up, we recorded 83,098 initial cardiovascular disease presentations. In each age group, the lowest risk for cardiovascular disease was in people with systolic blood pressure of 90-114 mm Hg and diastolic blood pressure of 60-74 mm Hg, with no evidence of a J-shaped increased risk at lower blood pressures. The effect of high blood pressure varied by cardiovascular disease endpoint, from strongly positive to no effect. Associations with high systolic blood pressure were strongest for intracerebral haemorrhage (hazard ratio 1·44 [95% CI 1·32-1·58]), subarachnoid haemorrhage (1·43 [1·25-1·63]), and stable angina (1·41 [1·36-1·46]), and weakest for abdominal aortic aneurysm (1·08 [1·00-1·17]). Compared with diastolic blood pressure, raised systolic blood pressure had a greater effect on angina, myocardial infarction, and peripheral arterial disease, whereas raised diastolic blood pressure had a greater effect on abdominal aortic aneurysm than did raised systolic pressure. Pulse pressure associations were inverse for abdominal aortic aneurysm (HR per 10 mm Hg 0·91 [95% CI 0·86-0·98]) and strongest for peripheral arterial disease (1·23 [1·20-1·27]). People with hypertension (blood pressure ≥140/90 mm Hg or those receiving blood pressure-lowering drugs) had a lifetime risk of overall cardiovascular disease at 30 years of age of 63·3% (95% CI 62·9-63·8) compared with 46·1% (45·5-46·8) for those with normal blood pressure, and developed cardiovascular disease 5·0 years earlier (95% CI 4·8-5·2). Stable and unstable angina accounted for most (43%) of the cardiovascular disease-free years of life lost associated with hypertension from index age 30 years, whereas heart failure and stable angina accounted for the largest proportion (19% each) of years of life lost from index age 80 years. The widely held assumptions that blood pressure has strong associations with the occurrence of all cardiovascular diseases across a wide age range, and that diastolic and systolic associations are concordant, are not supported by the findings of this high-resolution study. Despite modern treatments, the lifetime burden of hypertension is substantial. These findings emphasise the need for new blood pressure-lowering strategies, and will help to inform the design of randomised trials to assess them. Medical Research Council, National Institute for Health Research, and Wellcome Trust.


Immunological Findings and Clinical Outcomes of Infants With Positive Newborn Screening for Severe Combined Immunodeficiency From a Tertiary Care Center in the U.S.
The implementation of severe combined immunodeficiency (SCID) newborn screening has played a pivotal role in identifying these patients early in life as well as detecting various milder forms of T cell lymphopenia (TCL). In this study we reviewed the diagnostic and clinical outcomes, and interesting immunology findings of term infants referred to a tertiary care center with abnormal newborn SCID screens over a 6-year period. Key findings included a 33% incidence of non-SCID TCL including infants with novel variants in FOXN1, TBX1, MYSM1, POLD1, and CD3E; 57% positivity rate of newborn SCID screening among infants with DiGeorge syndrome; and earlier diagnosis and improved transplant outcomes for SCID in infants diagnosed after compared to before implementation of routine screening. Our study is unique in terms of the extensive laboratory workup of abnormal SCID screens including lymphocyte subsets, measurement of thymic output (TREC and CD4TE), and lymphocyte proliferation to mitogens in nearly all infants. These data allowed us to observe a stronger positive correlation of the absolute CD3 count with CD4RTE than with TREC copies, and a weak positive correlation between CD4RTE and TREC copies. Finally, we did not observe a correlation between risk of TCL and history of prenatal or perinatal complications or low birth weight. Our study demonstrated SCID newborn screening improves disease outcomes, particularly in typical SCID, and allows early detection and discovery of novel variants of certain TCL-associated genetic conditions.


Perception of self motion during and after passive rotation of the body around an earth-vertical axis.
We investigated the perception of self-rotation using constant-velocity chair rotations. Subjects signalled self motion during three independent tasks (1) by pushing a button when rotation was first sensed, when velocity reached a peak, when velocity began to decrease, and when velocity reached zero, (2) by rotating a disc to match the perceived motion of the body, or (3) by changing the static position of the dial such that a bigger change in its position correlated with a larger perceived velocity. All three tasks gave a consistent quantitative measure of perceived angular velocity. We found a delay in the time at which peak velocity of self-rotation was perceived (2-5 s) relative to the beginning or to the end of chair rotation. In addition the decay of the perception of self-rotation was preceded by a sensed constant-velocity interval or plateau (9-14 s). This delay in the rise of self-motion perception, and the plateau for the maximum perceived velocity, contrasts with the rapid rise and the immediate decay of the angular vestibuloocular reflex (aVOR). This difference suggests that the sensory signal from the semicircular canals undergoes additional neural processing, beyond the contribution of the velocity-storage mechanism of the aVOR, to compute the percept of self-motion.


Overexpression of EZH2 in multiple myeloma is associated with poor prognosis and dysregulation of cell cycle control.
Myeloma is heterogeneous at the molecular level with subgroups of patients characterised by features of epigenetic dysregulation. Outcomes for myeloma patients have improved over the past few decades except for molecularly defined high-risk patients who continue to do badly. Novel therapeutic approaches are, therefore, required. A growing number of epigenetic inhibitors are now available including EZH2 inhibitors that are in early-stage clinical trials for treatment of haematological and other cancers with EZH2 mutations or in which overexpression has been correlated with poor outcomes. For the first time, we have identified and validated a robust and independent deleterious effect of high EZH2 expression on outcomes in myeloma patients. Using two chemically distinct small-molecule inhibitors, we demonstrate a reduction in myeloma cell proliferation with EZH2 inhibition, which leads to cell cycle arrest followed by apoptosis. This is mediated via upregulation of cyclin-dependent kinase inhibitors associated with removal of the inhibitory H3K27me3 mark at their gene loci. Our results suggest that EZH2 inhibition may be a potential therapeutic strategy for the treatment of myeloma and should be investigated in clinical studies.


TTC7B is a new prognostic biomarker in head and neck squamous cell carcinoma linked to immune infiltration and ferroptosis.
To investigate the expression of TTC7B and its prognostic significance, biological roles, and impact on the immune system in patients with head and neck squamous cell carcinoma (HNSCC). Clinical and genomic data were obtained from TCGA (The Cancer Genome Atlas), GEO (Gene Expression Omnibus), GEPIA2 (Gene Expression Profiling Interactive Analysis 2.0), and TIMER2.0 (Tumor Immune Estimation Resource 2.0) databases. R software was utilized to process the retrieved data. qPCR and immunohistochemical assays were performed to validate the findings obtained from the databases. High expression of TTC7B was observed in HNSCC, and this heightened expression is significantly associated with reduced overall survival (OS) in patients, making it an independent risk factor impacting OS. TTC7B is correlated with focal adhesions and cell migration pathways based on functional enrichment analysis. CIBERSORT analysis and TIMER2.0 show a positive link between TTC7B and multiple immune cells, particularly macrophages. Pearson's analysis reveals a significant correlation between TTC7B and ferroptosis-related genes. In all, TTC7B could serve as a promising prognostic indicator of HNSCC, and is closely associated with focal adhesions, immune infiltration, and ferroptosis.


Advancing scholarship by publishing curriculum as an e-book.
Medical educators provide service by developing curricula and writing learning material. In addition, academic institutions expect medical educators to publish scholarship to be considered for promotion and academic advancement. Unfortunately, educators may receive limited time to execute these duties and expectations. One way medical educators can streamline their workload is by publishing educational coursework they have previously written into an e-book through an online publisher. This allows them to transform educational service they have already completed into scholarship required for academic recognition, thus maximizing the efficient use of their time. Publishing educational material as an e-book requires four steps. First, medical educators must determine which of their educational materials is best suited for publishing as an e-book. Second, educators must rank the features of each e-book publisher and choose the one that best meets their needs. Third, the educational material must be adapted as a manuscript and submitted for publication. Finally, the e-book must be advertised, promoted, and distributed to its intended audience. In addition, the success of the project should be evaluated. To illustrate this process, we describe the steps we took to publish the learning material we created for our internal medicine residents into an e-book. The overall process took approximately 3 months and went smoothly. For future publications, we would determine better ways to track the number of downloads of the e-book, ensure all of our images are adequately large, and consider the use of academic, rather than commercial e-book publishers.


Third-year Australian nursing students' attitudes, experiences, knowledge, and education concerning end-of-life care.
Nurses have a vital role in providing end-of-life care to patients and their families, and it is important that nursing students are adequately prepared for this role. This article reports on qualitative findings from research that explored a cohort of third-year undergraduate Australian nursing students' attitudes, experiences, knowledge, and education concerning end-of-life care. The study used open-ended questions in a purpose-designed, self-administered questionnaire and thematic analysis of the responses. Five themes emerged from the analysis: the importance of the students' values and beliefs, the influence of experience, their views on what constitutes a good or bad death, their knowledge of ethics and legislation surrounding end-of-life care, and how they perceived their level of education and knowledge. The need for more education on end-of-life care has implications for curriculum development in undergraduate nursing programmes, which need to provide graduating nurses with the necessary knowledge and skills to deliver quality care to patients who are dying and their families.


Sequence of complementary deoxyribonucleic acid encoding bonnet monkey (Macaca radiata) zona pellucida glycoprotein-ZP1 and its high-level expression in Escherichia coli.
Zona pellucida (ZP) glycoproteins have been proposed as candidate antigens for immunocontraception. Studies on this potential use can be facilitated by the availability of recombinant proteins. A cDNA lambda gt11 library was constructed using poly(A)+ mRNA isolated from bonnet monkey (Macaca radiata) ovaries and was screened for bonnet monkey ZP1 using a 404-basepair (bp) human ZP1 fragment (nucleotides 818-1221) as probe. Bonnet monkey ZP1 cDNA comprises 1617 nucleotides and encodes a polypeptide of 539 amino acid residues that share 92.0% identity with human ZP1. The major difference between bonnet monkey ZP1 and human ZP1 is the deletion of a 28-amino acid domain (amino acid residues 100-127 corresponding to human ZP1). An internal fragment (1317 bp) of bonnet monkey ZP1, excluding the N-terminus signal sequence and the C-terminus transmembrane-like domain, was amplified by polymerase chain reaction. The amplified Sac I and Kpn I restricted fragment was cloned in a frame downstream of the T5 promoter under the lac operator control for expression in the pQE-30 vector. Recombinant ZP1 (r-ZP1) was expressed as a polyhistidine fusion protein in Escherichia coli strains SG13009[pREP4] and ompT and Ion protease-deficient BL21 (plysS). SDS-PAGE analysis and immunoblotting with a murine monoclonal antibody, MA-410 (raised against porcine ZP3alpha--a homologue of bonnet monkey ZP1--and cross-reactive with bonnet monkey zona pellucida), revealed major bands of 51 and 40 kDa besides truncated fragments. Optimum expression of r-ZP1 was observed at 0.5 mM isopropyl beta-D-thiogalactopyranoside (IPTG). Immunization of male rabbits with r-ZP1 purified on nickel-nitrilotriacetic acid (NTA) resin under denaturing conditions and of female rabbits with r-ZP1 conjugated with diphtheria toxoid-generated antibodies reactive with r-ZP1 in ELISA. Moreover, immune sera, when tested by indirect immunofluorescence on bonnet monkey ovarian sections, showed positive fluorescence with zona pellucida. The information on the sequence of bonnet monkey ZP1 and the availability of the recombinant protein will help toward better understanding and evaluation of the contraceptive potential of homologous immunization in a nonhuman primate model.


How voluntary orienting of attention and alerting modulate costs of conflict processing.
There is evidence that pre-cued valid orienting of attention to competing information diminishes costs of brain processing of conflict. Still unclear, because scantily addressed by neuroimaging studies and mostly analyzed by means of behavioral indexing, it is whether conflict undergoes an equivalent modulation by tonic and pre-cued phasic alerting. Here, we investigated the functional relationships between attention orienting, alerting and executive systems using the Attention Network Test (ANT). Both reaction times (RTs) and ERPs were recorded. In line with previous literature, results showed that both RTs and a so-called ERPs conflict negativity (CN), prominent at anterior scalp and indexing conflict processing, were positively modulated by a prior valid orienting of attention onto the location of conflicting stimuli. Indeed, in this condition both kinds of markers showed faster latencies, while CN also reached higher amplitude values than in both alerting conditions, and, in turn, in pre-cued phasic alerting than in uncued tonic alerting. Moreover, while CN was larger over the right hemisphere independent of functional conditions, it was strongly modulated by the latter over the left hemisphere. Our ERP findings support the views of conflict modulation by both orienting and phasic alerting and of a functional integration between attentional brain networks.


Labetalol, nebivolol, and propranolol relax human radial artery used as coronary bypass graft.
Beta-blockers are a heterogeneous class of agents that are used in the treatment of many cardiovascular diseases, especially hypertension and atherosclerosis, and that are commonly prescribed after cardiac surgery. In the present study, the aim is to investigate the vasorelaxant effects of some common beta-adrenoceptor blockers on the human radial artery in vitro, as well as their relaxation mechanisms. Radial artery rings sourced from human patients were mounted in an organ bath and tested for changes in isometric tension in relaxation response to labetalol, nebivolol, and propranolol in the presence and absence of NG-nitro-L-arginine methyl ester (3 × 10(-5) mol/L) and tetraethyl ammonium (3 × 10(-4) mol/L). The labetalol (10(-8) to 10(-4) mol/L), nebivolol (10(-8) to 10(-4) mol/L), and propranolol (10(-8) to 10(-4) mol/L) induced concentration-dependent relaxations on the radial artery rings, which had been precontracted with phenylephrine (10(-6) mol/L). The relaxation response induced by labetalol in the isolated radial artery rings was significantly higher when compared with the nebivolol and propranolol samples (P < .05). NG-nitro-L-arginine methyl ester significantly reduced the relaxation of nebivolol (P < .05), and tetraethyl ammonium significantly reduced the relaxation of labetalol, nebivolol, and propranolol (P < .05). We speculated that the relaxant effect of labetalol, nebivolol, and propranolol was due partly to the Ca(2+)-activated K(+) channels. In addition, the relaxation induced by nebivolol was largely related with nitric oxide release. Nebivolol, and partly propranolol, may provide significant therapeutic benefit, but labetalol can be a good alternative for coronary artery bypass grafting with radial artery use.


Perceived quality of life of adults with developmental and other significant disabilities.
The purpose of this qualitative study was to describe the perceived quality of life (PQOL) of adults with significant disabilities who were living in community settings. Six male and six female volunteers (mean age 48 years, standard deviation 14 years), half with developmental and half with acquired disabilities, were interviewed about their quality of life. The interviews were audio-taped and transcribed verbatim. Text units were identified in the narrative data and were coded, sorted, and entered in an Index Tree. Data analysis utilized the Constant Comparison Technique with underlying themes emerging in 11 areas. The highest PQOL was noted in Health and Housing; the lowest in Transportation/Physical Accessibility and Employment. Deficits in Transportation/Physical Accessibility compromised PQOL in other areas. In regards to employment, seasonal, part-time and occasional work were common, and assistive technology and computers were rarely used to enhance employment capabilities. PQOL depended on the availability of appropriate supports and services, and tended to differ between those with developmental and those with acquired disabilities. Policy makers and advocacy groups could best improve the PQOL for those with severe disabilities by facilitating transportation and employment services.


Mg2+, Asp-, and Glu--effects in the processive and distributive DNA relaxation catalyzed by a eukaryotic topoisomerase.
The salt requirement for the catalysis of DNA relaxation carried out by a eukaryotic DNA topoisomerase I from Candida was reexamined with plasmid pBR322 DNA. Two levels of analysis were considered: the initial velocity of the overall reaction and the mode of this reaction (processivity vs distributivity). When looking at the monovalent salts from the first level, the replacement of Cl- by Glu- or Asp- greatly enhanced the salt range over which the enzyme was active. Moreover, the initial velocity reached an optimal value for a higher salt concentration in this case. For the cationic counterpart, K+ was a little more effective than Na+ and much more so than NH4+. Addition of 4 mM magnesium chloride affected both the range and the optimum of the initial velocity differentially, depending upon the monovalent salt, but with a general stimulating tendency. On the other hand, when the Mg2+ salt was varied, substitution of chloride by aspartate enhanced the optimum of the initial velocity for a fixed KCl concentration. In addition, magnesium aspartate (MgAsp2) and magnesium glutamate (MgGlu2) allowed the reaction to occur even without monovalent salt and over an extended range. Magnesium was also shown to directly interact with the general catalysis (Kd = 2.5 mM). From the second level of analysis, the presence of Mg2+ (except with NH4Glu), the substitution of Cl- by Glu- or Asp-, and a lower monovalent salt concentration than that used for the velocity optimum were required to promote the processive mode.(ABSTRACT TRUNCATED AT 250 WORDS)


Multiple ligand-ion solutions: a guide for solution preparation and computer program understanding.
This paper first gives an overview of the multitude of chemical reactions taking place in aqueous solutions with multiple ligands and ions. Then, recommendations to avoid pitfalls in preparing such solutions in daily laboratory work are summarized. Thereafter, the theoretical aspects for calculating multiple ligand-ion solutions are described in detail. Information necessary to perform corrections of the absolute stability constants for the influences of pH, ionic strength, and temperature are given. The calculation of apparent reaction constants for ligand-ion interactions by using the corrected absolute stability constants and considering all possible reactions between a ligand and an ion in an aqueous solution will be introduced. Thereafter, by employing the apparent reaction constants and an iterative method, the determination of multiple ligand-ion equilibria to find the desired ligand or ion concentrations are presented. Thus, this paper can be used as a guide to the principles of programs calculating multiple ligand-ion solutions, to choosing the appropriate program for one's special requirements, and to preparing multiple ligand-ion solutions properly.


Diagnostic accuracy study of the clinical indicators of vascular trauma in patients undergoing antineoplastic chemotherapy in peripheral veins.
Antineoplastic chemotherapy patients are susceptible to vascular trauma. The identification of this problem is possible through accurate clinical indicators. However, there are few diagnostic accuracy studies of vascular trauma in these patients. Thus, the objective was to analyze the accuracy of clinical indicators of vascular trauma in antineoplastic chemotherapy patients. A diagnostic accuracy study was carried out with a sample of 200 patients undergoing antineoplastic chemotherapy, in an oncology reference clinic, during 2018. A data collection form was created with sociodemographic and clinical data and indicators of vascular trauma. The sensitivity and specificity of the clinical indicators were assessed using a latent class analysis of random effects. The clinical indicators of decreased vascular elasticity (0.8384), pain (0.9573), and signs of infection at the catheter insertion site (0.9999) were specific for identifying vascular trauma in antineoplastic chemotherapy patients. The prevalence of vascular trauma in these patients was 11.17%. A set of three clinical indicators was considered accurate and statistically significant for confirming vascular trauma. This study has provided accurate clinical indicators of vascular trauma in antineoplastic chemotherapy patients. These results can contribute to establishing interventions, thereby reducing costs and maximizing health outcomes in this population.


Boosting Triticeae crop grain yield by manipulating molecular modules to regulate inflorescence architecture: insights and knowledge from other cereal crops.
One of the challenges for global food security is to reliably and sustainably improve the grain yield of cereal crops. One solution is to modify the architecture of the grain-bearing inflorescence to optimize for grain number and size. Cereal inflorescences are complex structures, with determinacy, branching patterns, and spikelet/floret growth patterns that vary by species. Recent decades have witnessed rapid advancements in our understanding of the genetic regulation of inflorescence architecture in rice, maize, wheat, and barley. Here, we summarize current knowledge on key genetic factors underlying the different inflorescence morphologies of these crops and model plants (Arabidopsis and tomato), focusing particularly on the regulation of inflorescence meristem determinacy and spikelet meristem identity and determinacy. We also discuss strategies to identify and utilize these superior alleles to optimize inflorescence architecture and, ultimately, improve crop grain yield.


Removal of Particulate Contamination from Solid Surfaces Using Polymeric Micropillars.
This Research Article describes a novel method for removal of particulate contamination, loosely referred to as dust, from solid surfaces using polymeric micropillars. In this Research Article, we illustrate for the first time that polymeric microfibrils of controlled interfacial and geometrical properties can effectively remove micrometric and submicrometric contaminant particles from a solid surface without damaging the underlying substrate. Once these microfibrils are brought into contact with a contaminated surface, because of their their soft and flexible structure, they develop intimate contact with both the surface contaminants and the substrate. While these intrinsically nonsticky micropillars have minimal interfacial interactions with the substrate, we show that they produce strong interfacial interactions with the contaminant particles, granting the detachment of the particles from the surface upon retraction of the cleaning material. The origin and strength of the interfacial interactions at the interfaces between a contaminant particle and both the substrate and the cleaning materials are thoroughly discussed. Unlike flat substrates of the same material, using microfibrillar structures of controlled interfacial and geometrical properties also allows the elimination of the adsorbed particles from the contact interface. Here we demonstrate that by moving the adsorbed particles from the tip to the side of the fibrils and consequently removing them from the contact interface, polymeric microfibrils can clean all contaminant particles from the surface. The effects of the geometrical and interfacial properties of polymeric micropillars on removing the adsorbed particles from the tips of the pillars are fully discussed. This research is not only important in terms of introducing a novel method which can offer a new paradigm for thorough yet nondestructive cleaning of dust particles from solid surfaces, but also it is of fundamental significance for researchers with interests in exploiting the benefits offered by microstructured surfaces in development of interfacially active materials and devices.


Multifaceted modes of action of the anticancer probiotic Enterococcus hirae.
A deviated repertoire of the gut microbiome predicts resistance to cancer immunotherapy. Enterococcus hirae compensated cancer-associated dysbiosis in various tumor models. However, the mechanisms by which E. hirae restored the efficacy of cyclophosphamide administered with concomitant antibiotics remain ill defined. Here, we analyzed the multifaceted modes of action of this anticancer probiotic. Firstly, E. hirae elicited emigration of thymocytes and triggered systemic and intratumoral IFNγ-producing and CD137-expressing effector memory T cell responses. Secondly, E. hirae activated the autophagy machinery in enterocytes and mediated ATG4B-dependent anticancer effects, likely as a consequence of its ability to increase local delivery of polyamines. Thirdly, E. hirae shifted the host microbiome toward a Bifidobacteria-enriched ecosystem. In contrast to the live bacterium, its pasteurized cells or membrane vesicles were devoid of anticancer properties. These pleiotropic functions allow the design of optimal immunotherapies combining E. hirae with CD137 agonistic antibodies, spermidine, or Bifidobacterium animalis. We surmise that immunological, metabolic, epithelial, and microbial modes of action of the live E. hirae cooperate to circumvent primary resistance to therapy.


Induced extrusion of DNA from the capsid of herpes simplex virus type 1.
DNA-filled capsids (C capsids) of herpes simplex virus type 1 were treated in vitro with guanidine-HCl (GuHCl) and analyzed for DNA loss by sucrose density gradient ultracentrifugation and electron microscopy. DNA was found to be lost quantitatively from virtually all capsids treated with GuHCl at concentrations of 0.5 M or higher, while 0.1 M GuHCl had little or no effect. DNA removal from 0.5 M GuHCl-treated capsids was effected without significant change in the capsid protein composition, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, or in its structure, as judged by electron microscopy. Electron microscopic examination of capsids in the process of emptying showed that DNA was extruded from multiple, discrete sites which appeared to coincide with capsid vertices. DNA exited the capsid in the form of thick strands or fibers that varied in diameter from approximately 4 to 13 nm with preferred diameters of 7 and 11 nm. The fibers most probably correspond to multiple, laterally aligned DNA segments, as their diameters are nearly all greater than that of a single DNA double helix. The results suggest that GuHCl treatment promotes an alteration in the capsid pentons which allows DNA to escape locally. Hexons must be more resistant to this change, since DNA loss appears to be restricted to the pentons. The ability of GuHCl to cause loss of DNA from C capsids with no accompanying change in capsid morphology or protein composition suggests that penton sites may open transiently to permit DNA exist and then return to their original state.