A preliminary investigation of lung availability of cannabinoids by smoking marijuana or dabbing BHO and decarboxylation rate of THC- and CBD-acids.
Highly potent cannabis concentrates obtained by butane or by supercritical carbon dioxide-extraction are gaining popularity. These extracts called butane hash oil (BHO) with Δ9-tetrahydrocannabinolic acid A (THCA) contents above 60% are consumed by flash vaporization on a glowing titanium nail, followed by inhalation of the resulting vapor through a water pipe in a single puff - a technique referred to as "dabbing". We herein investigated the decarboxylation rate of THCA during artificial smoking of cannabis plant material and simulated dabbing, and the lung availability of Δ9-tetrahydrocannabinol (THC) which we define as the recovery of THC in the smoke and vapor condensates. Preliminary smoking and dabbing experiments were performed using an apparatus built in-house. Due to availability of cannabidiol (CBD)-rich hemp in Switzerland, we included a sample of CBD flowers in our experiments and investigated the decarboxylation and recovery of cannabidiolic acid (CBDA) and CBD, respectively. Decarboxylation of THCA and CBDA during combustion of the plant material and vaporization of the BHO, respectively, was complete. The high recovery of total THC (75.5%) by dabbing cannot be achieved by smoking marijuana. Lung availability ranged from 12% for mixed cannabis material with a rather low THC content, to approximately 19-27% for marijuana flowers, similar for THC in marijuana as for CBD in CBD-rich marijuana. In reality, when smoking a joint, further losses in recovery must be assumed by additional sidestream smoke. The rather high lung availability of THC via dabbing can explain the increased psychoactive and adverse effects associated with this new trend of cannabis consumption.


The Conscripted Curriculum and the Reproduction of Racial Inequalities in Contemporary U.S. Medical Education.
In their attempt to address racial disparities in the provision of healthcare, the U.S. medical profession has reproduced racial inequalities of their own. In this article, I draw upon interview data with medical educators and students to detail how medical educators routinely offload the instruction on the social underpinnings and consequences of race onto students, particularly students of color. I develop the concept of the conscripted curriculum to capture how students' social identities are utilized by educators in the professionalization process. While there are exceptions in curricular approaches, most educators create the conscripted curriculum by eliciting students to share their social experiences with race in the small group setting while only providing students with didactic material on biological understandings of race. As a result, students of color report experiencing more emotionally exhausting and unrewarded labor than their white peers, and educators further devalue the social implications of race for healthcare.


Improved characterization of trastuzumab deruxtecan with PTCR and internal fragments implemented in middle-down MS workflows.
Antibody-drug conjugates (ADCs) are highly complex proteins mainly due to the structural microvariability of the mAb, along with the additional heterogeneity afforded by the bioconjugation process. Top-down (TD) and middle-down (MD) strategies allow the straightforward fragmentation of proteins to elucidate the conjugated amino acid residues. Nevertheless, these spectra are very crowded with multiple overlapping and unassigned ion fragments. Here we report on the use of dedicated software (ClipsMS) and application of proton transfer charge reduction (PTCR), to respectively expand the fragment ion search space to internal fragments and improve the separation of overlapping fragment ions for a more comprehensive characterization of a recently approved ADC, trastuzumab deruxtecan (T-DXd). Subunit fragmentation allowed between 70 and 90% of sequence coverage to be obtained. Upon addition of internal fragment assignment, the three subunits were fully sequenced, although internal fragments did not contribute significantly to the localization of the payloads. Finally, the use of PTCR after subunit fragmentation provided a moderate sequence coverage increase between 2 and 13%. The reaction efficiently decluttered the fragmentation spectra allowing increasing the number of fragment ions characteristic of the conjugation site by 1.5- to 2.5-fold. Altogether, these results show the interest in the implementation of internal fragment ion searches and more particularly the use of PTCR reactions to increase the number of signature ions to elucidate the conjugation sites and enhance the overall sequence coverage of ADCs, making this approach particularly appealing for its implementation in R&D laboratories.


Effect of hyperoxic exposure during early development on neurotrophin expression in the carotid body and nucleus tractus solitarii.
Synaptic activity can modify expression of neurotrophins, which influence the development of neuronal circuits. In the newborn rat, early hyperoxia silences the synaptic activity and input from the carotid body, impairing the development and function of chemoreceptors. The purpose of this study was to determine whether early hyperoxic exposure, sufficient to induce hypoplasia of the carotid body and decrease the number of chemoafferents, would also modify neurotrophin expression within the nucleus tractus solitarii (nTS). Rat pups were exposed to hyperoxia (fraction of inspired oxygen 0.60) or normoxia until 7 or 14 days of postnatal development (PND). In the carotid body, hyperoxia decreased brain-derived neurotrophic factor (BDNF) protein expression by 93% (P = 0.04) after a 7-day exposure, followed by a decrease in retrogradely labeled chemoafferents by 55% (P = 0.004) within the petrosal ganglion at 14 days. Return to normoxia for 1 wk after a 14-day hyperoxic exposure did not reverse this effect. In the nTS, hyperoxia for 7 days: 1) decreased BDNF gene expression by 67% and protein expression by 18%; 2) attenuated upregulation of BDNF mRNA levels in response to acute hypoxia; and 3) upregulated p75 neurotrophic receptor, truncated tropomyosin kinase B (inactive receptor), and cleaved caspase-3. These effects were not observed in the locus coeruleus (LC). Hyperoxia for 14 days also decreased tyrosine hydroxylase levels by 18% (P = 0.04) in nTS but not in the LC. In conclusion, hyperoxic exposure during early PND reduces neurotrophin levels in the carotid body and the nTS and shifts the balance of neurotrophic support from prosurvival to proapoptotic in the nTS, the primary brain stem site for central integration of sensory and autonomic inputs.


Expression of intracellular transforming growth factor-beta1 in CD4+CD25+ cells in patients with systemic lupus erythematosus.
The CD4+CD25+ regulatory T (Treg) cells exert immunoregulatory functions in various autoimmune diseases, in part through transforming growth factor-beta1 (TGF-beta1), and can be expanded by TGF-beta1 stimulation in normal subjects. This study aimed to examine intrinsic TGF-beta1 expression and the response to TGF-beta1 stimulation of this CD4+CD25+ subset in patients with systemic lupus erythematosus (SLE). Flow cytometry with multicolor staining of CD4+, CD25+, and TGF-beta1 was used to quantify the percentage of CD4+CD25+ T cells in fresh peripheral blood and TGF-beta1-stimulated peripheral blood mononuclear cell (PBMC) cultures, and their corresponding intracellular TGF-beta1 expression. In fresh peripheral blood, we found that decreased percentages of CD4+CD25+/CD4+ in SLE patients were associated with disease activity and renal involvement. Intracellular TGF-beta1 expression of CD4+CD25+ cells was significantly elevated in SLE compared with matched controls (p<0.001). In addition, there was significant negative correlation between TGF-beta1 expression and percentage of CD4+CD25+ cells present (r = -0.432, p=0.004). Nevertheless, in ex vivo unstimulated PBMC cultures, the percentage and intracellular TGF-beta1 expression of CD4+CD25+ cells of SLE were normalized to the levels of the control group. In TGF-beta1-stimulated PBMC cultures, CD4+CD25+ cells and their intracellular TGF-beta1 expression were significantly increased (p<0.001), both in SLE and controls. Moreover, the increments in the percentage of CD4+CD25+ cells and intracellular TGF-beta1 expression by TGF-beta1 stimulation were comparable in SLE and controls, and were not significantly influenced by disease activity or renal involvement in SLE. CD4+CD25+ cells were deficient in peripheral blood but not impaired either in intrinsic TGF-beta1 expression or in response to TGF-beta1 stimulation in patients with SLE. This study suggests that TGF-beta1, by inducing CD4+CD25+ cells, has potential clinical application in treating SLE.


Unenhanced CT for the evaluation of acute ureteric colic: the essential pictorial guide.
Acute ureteric colic is a common emergency, often dealt with by the emergency physician or general practitioner and referred on to the urologist. Unenhanced CT of the kidneys, ureters and bladder (CTKUB) is the 'gold standard' imaging investigation for establishing a diagnosis and guiding management. An appreciation of the CTKUB signs, which support or refute a diagnosis of ureteric colic, is highly valuable to the clinician when making a urological referral, and to the urologist, who must make appropriate management plans. All salient diagnostic and supportive features of ureteric colic are carefully illustrated, as are important radiological mimics, with the objectives of educating and informing the non-radiologist. Ready access to the picture archive and communication system (PACS) allows all specialists involved to interpret the radiological report with the benefit of images. A stone within the ureter may not always be readily apparent. Soft tissue rim sign around a calcific focus is an important indicator of a ureteric stone, whereas a comet tail sign suggests a phlebolith (a calcified venous thrombosis), a radiological mimic of a ureteric stone. Numerous secondary signs of ureteric obstruction may be present including hydronephrosis and perinephric stranding, and can help to confirm the diagnosis. The relative diagnostic weighting of signs is discussed, and a checklist is provided to assist with interpretation. Unexpected alternative radiological diagnoses are also illustrated, which may have significant management consequences necessitating specialist referral.


Zizyphus lotus L. fruit attenuates obesity-associated alterations: in vivo mechanisms.
Zizyphus lotus L., ZL is a Mediterranean plant and widely consumed for its beneficial medicinal properties. Objective: We assessed the effects of ZL fruit on diet-induced obesity. Materials and methods: Male C57BL/6j mice were divided into three groups. Each group received either a standard diet or a high-fat diet, HFD (30% of palm oil, w/v) or a HFD-supplemented with ZL fruit powder (10%, w/w) for six weeks, followed by a six weeks period, in which animals were maintained on the HFD and ZL aqueous extract (1%, w/v). We measured plasma parameters and assessed the expression of key genes involved in energy metabolism and inflammation. Results: ZL fruit improved glycaemia, lipids concentrations and inflammation in obese mice. Discussion and conclusion: Our investigations showed that ZL fruit improved glucose tolerance, dyslipidaemia and fatty liver disease, but not the severity of HFD-induced obesity in mice.


Deep brain stimulation of the nucleus accumbens for the treatment of addiction.
Despite novel medications and other therapeutic strategies, addiction to psychotropic substances remains one of the most serious public health problems worldwide. In this review, beginning with an introduction of deep brain stimulation (DBS), we highlight the importance of the nucleus accumbens (NAc) in the context of the reward circuitry and addictive behavior. We will provide a short historic overview of other neurosurgical approaches to treat addiction and describe the experimental and preclinical data on DBS in addiction. Finally, we call attention to key ethical issues related to using DBS to treat addiction that are important for future research and the design of clinical trials.


Biodiversity funds and conservation needs in the EU under climate change.
Despite ambitious biodiversity policy goals, less than a fifth of the European Union's (EU) legally protected species and habitats show a favorable conservation status. The recent EU biodiversity strategy recognizes that climate change adds to the challenge of halting biodiversity loss, and that an optimal distribution of financial resources is needed. Here, we analyze recent EU biodiversity funding from a climate change perspective. We compare the allocation of funds to the distribution of both current conservation priorities (within and beyond Natura 2000) and future conservation needs at the level of NUTS-2 regions, using modelled bird distributions as indicators of conservation value. We find that funding is reasonably well aligned with current conservation efforts but poorly fit with future needs under climate change, indicating obstacles for implementing adaptation measures. We suggest revising EU biodiversity funding instruments for the 2014-2020 budget period to better account for potential climate change impacts on biodiversity.


High speed digital protein interaction analysis using microfluidic single molecule detection system.
The understanding of protein interaction dynamics is important for signal transduction research but current available techniques prove difficult in addressing this issue. Thus, using the microfluidic approach, we developed a digital protein analytical platform and methodology named MAPS (Microfluidic system Analyzing Protein in Single complex) that can measure the amount of target proteins and protein complexes at the digitally single molecule resolution. By counting protein events individually, this system can provide rough protein interaction ratios which will be critical for understanding signal transduction dynamics. In addition, this system only requires less than an hour to characterize the target protein sample, which is much quicker than conventional approaches. As a proof of concept, we have determined the interaction ratios of oncogenic signaling protein complexes EGFR/Src and EGFR/STAT3 before and after EGF ligand stimulation. To the best of our knowledge, this is the first time that the interaction ratio between EGFR and its downstream proteins has been characterized. The information from MAPS will be critical for the study of protein signal transduction quantitation and dynamics.


Shear stress stimulation of p130(cas) tyrosine phosphorylation requires calcium-dependent c-Src activation.
Fluid shear stress (flow) modulates endothelial cell function via specific intracellular signaling events. Previously we showed that flow activated ERK1/2 in an integrin-dependent manner (Takahashi, M., and Berk, B. C. (1996) J. Clin. Invest. 98, 2623-2631). p130 Crk-associated substrate (Cas), a putative c-Src substrate, was originally identified as a highly phosphorylated protein that is localized to focal adhesions and acts as an adapter protein. Recent reports have shown that Cas is important in cardiovascular development and actin filament assembly. Flow (shear stress = 12 dynes/cm(2)) stimulated Cas tyrosine phosphorylation within 1 min in human umbilical vein endothelial cells. Phosphorylation peaked at 5 min (3.5 +/- 0.7-fold) and was sustained to 20 min. Tyrosine phosphorylation of Cas was functionally important because flow stimulated association of Cas with Crk in a time- and force-dependent manner. Flow-mediated activation of c-Src, phosphorylation of Cas, and association of Cas with Crk were all inhibited by calcium chelation and pretreatment with the Src family-specific tyrosine kinase inhibitor PP1. To determine the role of c-Src in flow-stimulated phosphorylation of Cas, we transduced cells with adenovirus encoding kinase-inactive Src. Expression of kinase-inactive Src prevented flow-induced Cas tyrosine phosphorylation but not ERK1/2 activation. Calcium-dependent activation of c-Src and tyrosine phosphorylation of Cas defines a new flow-stimulated signal pathway, different from ERK1/2 activation. This pathway may be involved in focal adhesion remodeling and actin filament assembly.


Evaluation of biocompatibility, osteointegration and biomechanical properties of the new Calcemex® cement: An in vivo study.
The mixture of polymethylmethacrylate (PMMA) and β-tricalciumphospate (β-TCP) is the most widely used bone graft. Common features of bone cement are the biocompatibility, bioactivity, mechanical stability and ability to fuse with the host's bone tissue. However, there are still few studies that have evaluated these characteristics in vivo. Our study aims to acquire these parameters, using an animal model with functional characteristics similar to those of humans. The analyzed cement is Calcemex®, evaluated both in compact and fluid formulation. The chosen animal models were 5 pigs, treated with femoral and tibial implants of Calcemex® samples. After one year, the pigs were sacrificed and the specimens explanted for morphological, histological, ultrastructural and mechanical evaluations. For both formulations, the investigation highlighted the absence of foreign body reactions in the host, the histological integration with the surrounding tissues and the preservation of mechanical compression resistance.


An anticoagulation therapy elective for third-year pharmacy students.
To develop an anticoagulation elective course for third-year pharmacy students to enhance their knowledge and skills for providing anticoagulation services. Content developed for the course focused on standards of care, evaluation of primary literature, clinical application, billing and regulatory issues, written and verbal communication, and patient education. Teaching methods included lectures, discussions, demonstrations, and self-directed learning. Assessment methods included multiple-choice examinations, evaluation of patient cases, student presentations, and a practical examination. Students demonstrated competencies in multiple areas and rated the course favorably. Students who completed the elective reported being more prepared for anticoagulation activities than those who did not. Confidence levels regarding participation in anticoagulation services postgraduation were similar. The elective provided valuable experience in anticoagulation therapy and increased students' perception of their preparedness related to dealing with anticoagulation issues while on advanced pharmacy practice experiences (APPEs). More exposure to management topics and the logistics of initiating anticoagulation services should be incorporated.


[Monitoring of chemotherapy in patients with pulmonary tuberculosis by the blood bacteriostatic activity-determining test].
The paper gives the results of studies to determine blood bacteriostatic activity (BBA) in the use of a patient's autostrain and semiliquid medium versus the clinical and laboratory parameters of the course of a process in 101 patients with pulmonary tuberculosis. There is evidence for the relationship of the BBA to the sensitivity to isoniazid and the structure of drug resistance. The zero values of BBA correspond to the severest course of the disease. The efficiency of treatment is much higher in patients with high and moderate BBA. The latter's determination using the semiliquid medium permits an objective evaluation of the efficiency of chemotherapy, identification of patients with a poor prognosis, and then choice of an individual treatment regimen on day 7 after the test just before obtaining the data on drug sensitivity.


Conversations between persons with dementia disease living in nursing homes and nurses--qualitative evaluation of an intervention with the validation method.
Living with dementia disease (DD) can include difficulties describing experiences of everyday lives, which can lead to withdrawal, social isolation or existential homelessness. Persons with DD living in nursing homes are mainly dependent on the nurses for establishing and maintaining relationships with those around them. It can be challenging for nurses to understand what a person with DD is trying to express and to make themselves understood in turn. The validation method is intended to facilitate communication with persons with DD, but to our knowledge, there have been no qualitative studies of how this influences persons' communication. This study aimed to illuminate the actions and reactions of persons with DD living in nursing homes in one-to-one conversations with nurses during 1 year of validation method training, as observed in videotapes. Four persons with DD were involved in videotaped conversations with four nurses who were participating in a validation method training programme. Videotapes with at least 5 months between the first and last recording were analysed and compared qualitatively. The findings are presented in four categories that were identified to various degrees in conversations at the beginning and at the end of the programme: being uninterested in or unable to answer questions, talking about more than one topic of conversation at the same time, trying to talk about what is on one's mind and speaking more freely about what is on one's mind. In the videotaped conversations at the end of the programme, the persons had the opportunity to use their remaining communication abilities. This may have been related to the development of the nurses' communication skills during the training programme, and so it is possible that persons with DD could benefit from communicating with nurses trained in the validation method.


The direct thrombin inhibitor hirudin.
This review discusses the pharmacology and clinical applications of hirudin, a bivalent direct thrombin inhibitor (DTI). Besides the current major indication for hirudin--anticoagulation of patients with heparin-induced thrombocytopenia (HIT)--the experience with hirudin in other indications, especially acute coronary syndromes, are briefly presented. Hirudins have been formally studied prior to their regulatory approval; however, important information on their side effects and relevant preventative measures only became available later. Therefore, current recommendations and dosing schedules for hirudin differ considerably from the information given in the package inserts. Drawbacks of hirudin and important precautions for avoiding potential adverse effects are discussed in detail in the third part of this review.


Expression Profiling of Candidate Genes in Sugar Beet Leaves Treated with Leonardite-Based Biostimulant.
Leonardite-based biostimulants are a large class of compounds, including humic acid substances. Foliar application of biostimulants at field level improves plant growth, yield and quality through metabolic changes and stimulation of plant proton pumps. The present study aimed at identifying optimum dosage of BLACKJAK, a humic acid-based substance, which is able to modify genes involved in sugar beet growth. Thirty-three genes belonging to various biochemical pathway categories were tested in leaves of treated sugar beet (Beta vulgaris L.) samples to assess gene expression profiling in response to BLACKJAK. Seedlings of a diploid and multigerm variety were grown in plastic pots and sprayed with two dilutions of BLACKJAK (dilution 1:500-1.0 mg C L-1 and dilution 1:1000-0.5 mg C L-1). Leaf samples were collected after 24, 48, and 72 h treatment with BLACKJAK for each dilution. RNA was extracted and the quantification of gene expression was performed while using an OpenArray platform. Results of analysis of variance demonstrated that, 15 genes out of a total of 33 genes tested with OpenArray qPCR were significantly affected by treatment and exposure time. Analysis for annotation of gene products and pathways revealed that genes belonging to the mitochondrial respiratory pathways, nitrogen and hormone metabolisms, and nutrient uptake were up-regulated in the BLACKJAK treated samples. Among the up-regulated genes, Bv_PHT2;1 and Bv_GLN1 expression exerted a 2-fold change in 1:1000 and 1:500 BLACKJAK concentrations. Overall, the gene expression data in the BLACKJAK treated leaves demonstrated the induction of plant growth-related genes that were contributed almost to amino acid and nitrogen metabolism, plant defense system, and plant growth.


Bempedoic Acid (ETC-1002): ATP Citrate Lyase Inhibitor: Review of a First-in-Class Medication with Potential Benefit in Statin-Refractory Cases.
Bempedoic acid (BA; ETC-1002) is a new agent that reduces cholesterol synthesis through inhibition of adenosine triphosphate citrate lyase, an enzyme upstream from 3-hydroxy-3-methylglutaryl-coenzyme A. In animal models, BA also influences fatty acid synthesis, but in humans, its role is limited primarily to lowering low-density lipoprotein cholesterol (LDL-C). In early clinical trials, BA was well tolerated and without major side effects. Alone or in various combinations with atorvastatin and/or ezetimibe, LDL-C lowering ranged from 17% to 64%. In addition, BA lowers levels of non-high-density lipoprotein cholesterol, C-reactive protein, and apolipoprotein B. Statins are first-line agents for primary and secondary prevention of cardiovascular disease. However, muscle-related side effects and other problems such as elevated liver enzymes may limit their use. In addition, LDL-C lowering beyond that provided by statin therapy alone may be needed. BA may be useful in either of these scenarios, as it is relatively free of muscle-related side effects and appears to enhance LDL-C lowering beyond that achieved with statin monotherapy. Phase 3 trials and one outcomes study are currently under way to better define this agent's potential clinical role.


On the shoulders of giants: Harvey Cushing's experience with acromegaly and gigantism at the Johns Hopkins Hospital, 1896-1912.
A review of Dr. Cushing's surgical cases at Johns Hopkins Hospital revealed new information about his early operative experience with acromegaly. Although in 1912 Cushing published selective case studies regarding this work, a review of all his operations for acromegaly during his early years has never been reported. We uncovered 37 patients who Cushing treated with surgical intervention directed at the pituitary gland. Of these, nine patients who presented with symptoms of acromegaly, and one with symptoms of gigantism were selected for further review. Two patients underwent transfrontal 'omega incision' approaches, and the remaining eight underwent transsphenoidal approaches. Of the 10 patients, 6 were male. The mean age was 38.0 years. The mean hospital stay was 39.4 days. There was one inpatient death during primary interventions (10%) and three patients were deceased at the time of last follow-up (33%). The mean time to death, calculated from the date of the primary surgical intervention, and including inpatient and outpatient deaths, was 11.3 months. The mean time to last follow-up, calculated from the day of discharge, was 59.3 months. At the time of last follow-up, two patients reported resolution of headache; four patients reported continued visual deficits, and two patients reported ongoing changes in mental status. This review analyzes the outcomes for 10 patients who underwent surgical intervention for acromegaly or gigantism, and offers an explanation for Cushing's transition from the transfrontal "omega incision" to the transsphenoidal approach while practicing at the Johns Hopkins Hospital.


Experimental Evaluation of Hiṅgvādi Ghṛta in Behavioral Despair Using Animal Models.
Depression, a sustained mood disorder caused by selective diminution of specialized cells in brain is increasing at an alarming rate. It will be the second largest morbid illness by next decade and is the leading cause of suicidal deaths. The available antidepressant medications benefit only a third of its recipients and have many side effects. Hence, it is imperative to search in Ayurveda for leads. To evaluate Anti- depressant activity of Hiṅgvādi Ghṛta in vivo . Comparative preclinical study. Hiṅgvādi Ghṛta (HG) was prepared using standard operating procedure, physicochemically analyzed and assessed. Tail Suspension Test (TST) model with Swiss albino mice and Forced Swim Test (FST) model with Wistar albino rats were used to assess anti-depressant activity. Imipramine hydrochloride in dose of 15 mg/kg for TST and 10 mg/kg for FST, was the standard drug and Ghee as vehicle control in dose of 0.1g/20g for TST and 0.72g/200g for FST orally. Hiṅgvādi Ghṛta in doses of 0.05g/20g (x/2), 0.1g/20g (x) and 0.2 g/20g (2x) for TST and 0.36g/200g (x/2), 0.72g/200g (x) and 1.44g/200g (2x) for FST was administered to 3 test groups for 21 days orally except Plain control group which received only distilled water. Duration of immobility in seconds for TST and number of rotations for FST were noted for assessment. One way ANOVA followed by Dunnets test and Paired t test. HG was significantly effective at dose of 0.1gm/20gm for TST (P = 0.0037; P < 0.01) and 0.72g/200g for FST (P = 0.0055, P < 0.01) comparable to Imipramine hydrochloride. HG displayed potent anti depressant activity comparable to standard drug Imipramine Hydrochloride.


Identification of the viral haemorrhagic disease virus of rabbits as a calicivirus.
Liver tissue from rabbits which had died of viral haemorrhagic disease (VHD) was used to identify the causative agent of the disease. After extraction of liver homogenates and density gradient ultracentrifugation, viral particles with buoyant densities between 1.32 to 1.38 g/ml and estimated sedimentation coefficients between 100 S and 175 S were obtained. The isolated virions were determined to have a diameter of 40 nm. The particles morphologically resembled those of the Caliciviridae family, had positive reactions both in ELISA and haemagglutination assays, and were infective for rabbits. By immunoblotting, a major structural protein with a molecular weight of 60 kDa was identified. RNA of high purity and of approximately 8 kb was isolated from virions. Labelled cDNA of virion RNA detected two RNAs of 8 kb and 2 kb in Northern blots of liver RNA from animals infected with the VHD virus (VHDV). Finally, isolated virion RNA injected directly into the liver or rabbits produced a disease with clinical symptoms and pathological findings typical of VHD. A calicivirus originally designated "rabbit haemorrhagic disease virus (RHDV)" was thus identified as the causative agent of VHD. The agent causing the European brown hare syndrome (EBHS) is also a calicivirus (EBHSV). VHDV and EBHSV are different members of the Caliciviridae family; their relationship can be clearly demonstrated by using different approaches of investigation.


Breast Cancer Classification by Using Multi-Headed Convolutional Neural Network Modeling.
Breast cancer is one of the most widely recognized diseases after skin cancer. Though it can occur in all kinds of people, it is undeniably more common in women. Several analytical techniques, such as Breast MRI, X-ray, Thermography, Mammograms, Ultrasound, etc., are utilized to identify it. In this study, artificial intelligence was used to rapidly detect breast cancer by analyzing ultrasound images from the Breast Ultrasound Images Dataset (BUSI), which consists of three categories: Benign, Malignant, and Normal. The relevant dataset comprises grayscale and masked ultrasound images of diagnosed patients. Validation tests were accomplished for quantitative outcomes utilizing the exhibition measures for each procedure. The proposed framework is discovered to be effective, substantiating outcomes with only raw image evaluation giving a 78.97% test accuracy and masked image evaluation giving 81.02% test precision, which could decrease human errors in the determination cycle. Additionally, our described framework accomplishes higher accuracy after using multi-headed CNN with two processed datasets based on masked and original images, where the accuracy hopped up to 92.31% (±2) with a Mean Squared Error (MSE) loss of 0.05. This work primarily contributes to identifying the usefulness of multi-headed CNN when working with two different types of data inputs. Finally, a web interface has been made to make this model usable for non-technical personals.


Long-term continuous exercise training counteracts the negative impact of the menopause transition on cardiometabolic health in hypertensive women - a 9-year RCT follow-up.
The study examined effects of 9-yrs of multicomponent exercise training during the menopause interval on cardiometabolic health in hypertensive women. Sedentary, middle-aged women (n = 25) with mild-to-moderate arterial hypertension were randomized into a soccer training (multicomponent exercise; EX; n = 12) or control group (CON; n = 13). EX took part in 1-h football training sessions, 1-3 times weekly, for a consecutive 9-years, totaling ~800 training sessions, while CON did not take part in regular exercise training. 22 participants entered menopause during the intervention. A time×group interaction effect (P = 0.04) of 8.5 mmHg in favour of EX was observed for changes in mean arterial pressure (MAP) (EX: -4.8 [-10.7;1.1] mmHg, CON +3.7 [-2.0;9.3] mmHg). Time×group interaction effects in favour of EX were also observed for total body weight (4.6 kg, P = 0.008, EX: +0.7 [-1.7;3.0] kg, CON: +5.3 [3.0;7.6] kg, total fat percentage (5.7%-points, P = 0.02; EX (-1.9 [-4.4;0.6] %-points; P = 0.13), CON +3.8 [1.4;6.2] %-points and for total cholesterol (1.2 mmol/l, P = 0.03, EX: -0.5 [-1.0;-0.1] mmol/l, CON: +0.7 [0.2;1.1] mmol/l. EX reduced (P = 0.02) plasma low-density lipoprotein cholesterol by -0.4 [-0.8;-0.1] mmol/l, whereas an increase (P = 0.01) of 0.4 [0.1;0.8] mmol/l occurred in CON (interaction. P < 0.001). A time×group interaction (P = 0.004) existed for changes in exercise capacity in favour of EX. Fasting glucose remained unchanged in EX and increased (P < 0.001) by 0.7 [0.4;1.0] mmol/l in CON (time×group interaction P = 0.02). In conclusion, long-term multicomponent exercise training fully counteracts the detrimental effects of the menopause transition on cardiometabolic health in hypertensive women.


InsP3-dependent Ca2+ mobilization in circular but not longitudinal muscle cells of intestine.
The presence and functional coupling of inositol 1,4,5-trisphosphate (InsP3) receptors was examined in permeabilized muscle cells and microsomes isolated separately from the circular and longitudinal muscle layers of guinea pig intestine. Specific binding of [3H]InsP3 occurred only in circular muscle cells and microsomes; binding to longitudinal muscle cells or microsomes was minimal or absent. Binding to circular muscle cell microsomes was rapid (steady state in 2 min), saturable, reversible, and temperature independent, and was inhibited in a concentration-dependent manner by unlabeled InsP3 (IC50 10 nM) and by the InsP3 receptor antagonist heparin (EC50 4 micrograms/ml); inhibition by InsP4 and InsP2 was minimal. Consistent with the preferential location of InsP3 receptors, InsP3 elicited concentration-dependent contraction (EC50 3 nM), 45Ca2+ efflux (EC50 55 nM), and increase in [Ca2+]i in circular muscle cells and 45Ca2+ efflux in microsomes; all these events were blocked by heparin (10 micrograms/ml). Although ATP-dependent Ca2+ stores were present in longitudinal muscle cells and microsomes, they were not sensitive to 1 microM InsP3, which was maximally effective in circular muscle cells; 100 microM InsP3 elicited partial Ca2+ release (30-50%) and contraction (47%) that could only be blocked by high concentrations of heparin (100 micrograms/ml). We conclude that specific, high-affinity, heparin-sensitive InsP3 receptors capable of mobilizing intracellular Ca2+ are present predominantly in circular muscle cells of the intestine.


[Changes in the transmembrane potential of developing loach embryos under the influence of insulin, inhibition of transcription and translation].
Results of continuous 7-8 hour registration of transmembrane potential (TMP) at the early stages of the loach (Misgurnus fossilis) development in normal conditions and in media with different insulin concentrations (the hormone with actinomycin D and cycloheximid and these antibiotics alone) are presented in this paper. The dependence of the absolute value and the mode of TMP changes in developing embryos upon insulin concentration has been established. It is shown that actinomycin D reduces the hormonal effect at the initial stages, while alone it does not cause any visible changes of TMP. Inhibition of translation with cycloheximid leads to sharp depolarizaton of the membrane and completely abolishes the periodicity of TMP oscillations, which in the norm corresponds to the time-course of the mitotic cycles during the synchronous cleavang of the blastomeres.


Childhood abuse and criminal behavior: testing a general strain theory model.
This article draws on general strain theory (GST) to develop and test a model of the childhood abuse-crime relationship. Using data from the National Longitudinal Study of Adolescent Health (Add Health),(1) we find that early childhood physical and sexual abuse are robust predictors of offending in adolescence, for the full sample and in equations disaggregated by gender. GST is partially supported in that the effects of childhood physical abuse on offending for both females and males are mediated by an index of depression symptoms, whereas the effect of sexual abuse among females appears to be mediated largely by closeness to mother. The effect of childhood sexual abuse among males, however, is more robust than among females and it persists despite controls for low self-control, ties to delinquent peers, school attachment, and closeness to mother. Theoretical implications of the findings are discussed.


Managing the analytical challenges related to micro- and nanoplastics in the environment and food: filling the knowledge gaps.
This paper identifies knowledge gaps on the sustainability and impacts of plastics and presents some recommendations from an expert group that met at a special seminar organised by the European Commission at the end of 2018. The benefits of plastics in society are unquestionable, but there is an urgent need to better manage their value chain. The recently adopted European Strategy for Plastics stressed the need to tackle the challenges related to plastics with a focus on plastic litter including microplastics. Microplastics have been detected mainly in the marine environment, but also in freshwater, soil and air. Based on today's knowledge they may also be present in food products. Although nanoplastics have not yet been detected, it can be assumed that they are also present in the environment. This emerging issue presents challenges to better understand future research needs and the appropriate immediate actions to be taken to support the necessary societal and policy initiatives. It has become increasingly apparent that a broad and systematic approach is required to achieve sustainable actions and solutions along the entire supply chain. It is recognised that there is a pressing need for the monitoring of the environment and food globally. However, despite the number of research projects increasing, there is still a lack of suitable and validated analytical methods for detection and quantification of micro- and nanoplastics. There is also a lack of hazard and fate data which would allow for their risk assessment. Some priorities are identified in this paper to bridge the knowledge gaps for appropriate management of these challenges. At the same time it is acknowledged that there is a great complexity in the challenges that need to be tackled before a really comprehensive environmental assessment of plastics, covering their entire life cycle, will be possible.


P2X7 Deficiency Blocks Lesional Inflammasome Activity and Ameliorates Atherosclerosis in Mice.
Extracellular adenosine triphosphate (ATP) binds as a danger signal to purinergic receptor P2X7 and promotes inflammasome assembly and interleukin-1β expression. We hypothesized a functional role of the signal axis ATP-P2X7 in inflammasome activation and the chronic inflammation driving atherosclerosis. P2X7-competent and P2X7-deficient macrophages were isolated and stimulated with lipopolysaccharide, ATP, or both. To assess whether P2X7 may have a role in atherosclerosis, P2X7 expression was analyzed in aortic arches from low density lipoprotein receptor-/- mice consuming a high-cholesterol or chow diet. P2X7+/+ and P2X7-/- low density lipoprotein receptor-/- mice were fed a high-cholesterol diet to investigate the functional role of P2X7 knockout in atherosclerosis. Human plaques were derived from carotid endarterectomy and stained against P2X7. Lipopolysaccharide or ATP stimulation alone did not activate caspase 1 in isolated macrophages. However, priming with lipopolysaccharide, followed by stimulation with ATP, led to an activation of caspase 1 and interleukin-1β in P2X7-competent macrophages. In contrast, P2X7-deficient macrophages showed no activation of caspase 1 after sequential stimulation while still expressing a basal amount of interleukin-1β. P2X7 receptor was higher expressed in murine atherosclerotic lesions, particularly by lesional macrophages. After 16 weeks of a high-cholesterol diet, P2X7-deficient mice showed smaller atherosclerotic lesions than P2X7-competent mice (0.162 cm2±0.023 [n=9], P2X7-/- low density lipoprotein receptor-/- : 0.084 cm2±0.01 [n=11], P=0.004) with a reduced amount of lesional macrophages. In accord with our in vitro findings, lesional caspase 1 activity was abolished in P2X7-/- mice. In addition, intravital microscopy revealed reduced leukocyte rolling and adhesion in P2X7-deficient mice. Last, we observe increased P2X7 expression in human atherosclerotic lesions, suggesting that our findings in mice are relevant for human disease. P2X7 deficiency resolved plaque inflammation by inhibition of lesional inflammasome activation and reduced experimental atherosclerosis. Therefore, P2X7 represents an interesting potential new target to combat atherosclerosis.


DNAJC30 biallelic mutations extend mitochondrial complex I-deficient phenotypes to include recessive Leber's hereditary optic neuropathy.
Leber's hereditary optic neuropathy (LHON) is the most common mitochondrial disease and in most cases is caused by mutations in mitochondrial DNA-encoded (mtDNA-encoded) respiratory complex I subunit ND1, ND4, or ND6. In this issue of the JCI, Stenton et al. describe biallelic mutations in a nuclear DNA-encoded gene, DNAJC30, establishing recessively inherited LHON (arLHON). Functional studies suggest that DNAJC30 is a protein chaperone required for exchange of damaged complex I subunits. Hallmark mtDNA LHON features were also found in arLHON, including incomplete penetrance, male predominance, and positive response to idebenone therapy. These results extend complex I-deficient phenotypes to include recessively inherited optic neuropathy, with important clinical implications for genetic counseling and therapeutic considerations.


Bodies in motion: Unraveling the distinct roles of motion and shape in dynamic body responses in the temporal cortex.
The temporal cortex represents social stimuli, including bodies. We examine and compare the contributions of dynamic and static features to the single-unit responses to moving monkey bodies in and between a patch in the anterior dorsal bank of the superior temporal sulcus (dorsal patch [DP]) and patches in the anterior inferotemporal cortex (ventral patch [VP]), using fMRI guidance in macaques. The response to dynamics varies within both regions, being higher in DP. The dynamic body selectivity of VP neurons correlates with static features derived from convolutional neural networks and motion. DP neurons' dynamic body selectivity is not predicted by static features but is dominated by motion. Whereas these data support the dominance of motion in the newly proposed "dynamic social perception" stream, they challenge the traditional view that distinguishes DP and VP processing in terms of motion versus static features, underscoring the role of inferotemporal neurons in representing body dynamics.


Prevalence and correlates of depression among black and Latino stroke survivors with uncontrolled hypertension: a cross-sectional study.
To examine the prevalence and correlates of depression in a cohort of black and Hispanic stroke survivors with uncontrolled hypertension. Baseline survey data from 10 stroke centres across New York City. Black and Hispanic stroke survivors with uncontrolled hypertension (n=450). Depressive symptoms were assessed with the 8-item Patient Reported Outcomes Measurement Information System (PROMIS) measure. Depression was defined as a PROMIS score ≥55. Other data collected included clinical factors, health-related quality of life (EuroQoL five dimensions (EQ-5D)), functional independence (Barthel Index, BI), stroke-related disability (Modified Rankin Score), physical function (PROMIS Physical Function) and executive functioning (Frontal Assessment Battery). The mean age was 61.7±11.1 years, 44% of participants were women and 51% were black. Poststroke depression was noted in 32% of the cohort. Examining bivariate relationships, patients with depression were observed to have poorer function and quality of life as evidenced by significantly lower PROMIS physical function scores (36.9±8.32 vs 43.4±10.19, p<0.001); BI scores (79.9±19.2 vs 88.1±15.1, p<0.001); EQ-5D scores (0.66±0.24 vs 0.83±0.17, p<0.001) and higher Rankin scores (2.10±1.00 vs 1.46±1.01, p<0.001) compared with those without depression. Multivariate (model adjusted) significant correlates of depression included lower self-reported quality of life (OR=0.02 (CI 0.004 to 0.12) being younger (OR=0.94; 95% CI 0.91 to 0.97); not married (OR=0.46; CI 0.24 to 0.89)); and foreign-born (OR=3.34, 95% CI 1.4 to 7.97). There was a trend for higher comorbidity to be uniquely associated with depression (≥3 comorbid conditions, OR=1.49, 95% CI 1.00 to 2.23). Poststroke depression is common among black and Hispanic stroke survivors with higher rates noted among foreign-born patients and those with high comorbidity. These findings highlight the importance of screening for depression in minority stroke survivors. http://www.clinicaltrials.gov. Unique identifier: NCT01070056.


They Care for Others, But What About Themselves? Understanding Self-Care Among DSPs' and Its Relationship to Professional Quality of Life.
Direct support professionals (DSPs) are instrumental to the daily operations of organizations that support people with intellectual and developmental disabilities (IDD). With extensive responsibilities, DSPs often experience high levels of stress and burnout that can result in turnover and vacant positions. Self-care is the practice of behaviors that promote well-being, counter work-related stress, and foster resilience. The current study explored self-care and resilience, and their relationship with professional quality of life (i.e., satisfaction, burnout, and secondary traumatic stress) among DSPs. Using a convenient sample, 153 DSPs (71% female) completed an online survey comprised of multiple measures. Results indicated that DSPs often engaged in self-care behaviors across physical, psychological, emotional, spiritual, relational, and workplace domains, however, less than 40% engaged in self-care behaviors directly related to work. On average, DSPs reported high levels of resilience. Collectively, self-care and resilience accounted for 12% to 28% of variance in DSPs' professional quality of life. Given the contribution of self-care to resilience and professional quality of life, an active approach by IDD organizations to foster self-care among DSPs may help promote their longevity and retention.


Depositing light in a photonic stop gap by use of Kerr nonlinear microresonators.
We show theoretically that it is possible to trap light in a microresonator structure by use of four-wave mixing. The efficiency of the parametric process is substantially increased by the high group delay of light inside the structure. The energy that is trapped has a half-life of approximately 500 ps in the presence of both linear and nonlinear loss in the channel waveguides and resonators. We also demonstrate that the energy can be extracted from the cavity with a similar process.


Polyherbal formulation Anoac‑H suppresses the expression of RANTES and VEGF for the management of bleeding hemorrhoids and fistula.
Hemorrhoids and fistula are considered the most common anorectal conditions in the general population. These conditions affect the quality of a patient's life by causing pain and bleeding during defecation or even in the resting state. Lower grades of hemorrhoids may be controlled by traditional measures. However, surgery is an effective treatment option in recurrent‑lower and higher‑grade hemorrhoids. Surgical procedures are associated with various complications, including pain and delayed wound healing. Recurrence of hemorrhoids is also a major concern in the post‑operative period. An anal fistula is the connection between the anus and the skin and causes severe pain, swelling, as well as blood and pus discharge. Fistula has serious social and economic consequences. Hence, it is important to understand the pathophysiology and molecular pathology of hemorrhoids and fistula, to identify the molecular targets and to develop pharmacological‑interventions. In a previous study by our group, the polyherbal formulation Anoac‑H was developed for the treatment of different stages of hemorrhoids and fistula, and it was demonstrated that Anoac‑H is an effective formulation for treating hemorrhoids. However, the molecular mode of action of Anoac‑H on hemorrhoids and fistula had remained elusive. In the present study, it was determined that this formulation reduces the migration of mesenchymal (fibroblasts) and immune (RAW 264.7) cells without affecting their viability. It was also observed that Anoac‑H suppresses the expression of regulated upon activation, normal T cell expressed and presumably secreted (RANTES) and VEGF in fibroblasts and macrophages. Inflammation and elevated expression of RANTES and VEGF were observed in hemorrhoids and fistula. However, inflammation, as well as the expression of RANTES and VEGF, were significantly reduced in treated human hemorrhoid and fistula tissues as compared to untreated ones, confirming the in vitro results.


Approach to the patient: contraception in women with polycystic ovary syndrome.
Polycystic ovary syndrome (PCOS) is a common reproductive and metabolic disorder. Patients with PCOS present with clinical signs of androgen excess (ie, hirsutism and acne), menstrual irregularities, and infertility. Combined oral contraceptive (OC) pills are the first-line medical therapy for the long-term management of PCOS. Containing a combination of estrogen and progestin, OCs restore regular menses, improve androgen excess, and provide effective contraception and protection from endometrial cancer. The benefits of hormonal contraception outweigh the risks in the vast majority of women with PCOS. However, concerns have been raised about potential adverse cardiovascular and metabolic effects of OCs. Currently available evidence indicates an increased relative risk of venous thrombosis associated with OCs varying among different formulations. Arterial thrombosis risk attributable to OCs does not appear to be significantly increased in young nonsmoking women. OC use might be associated with increased risk of diabetes in morbidly obese women with PCOS with severe insulin resistance. A tailored clinical approach to oral contraception in women with PCOS requires individualized risk stratification and management by determination of each PCOS patient's personal cardiometabolic risk profile at baseline and during follow-up. Before prescribing an OC, clinicians should document individual risk factors including age, smoking, obesity, any degree of glucose intolerance including prediabetes and diabetes, hypertension, dyslipidemia, thrombophilia, and personal or family history of a venous thromboembolic event.


Discovery and characterization of distinct simian pegiviruses in three wild African Old World monkey species.
Within the Flaviviridae, the recently designated genus Pegivirus has expanded greatly due to new discoveries in bats, horses, and rodents. Here we report the discovery and characterization of three simian pegiviruses (SPgV) that resemble human pegivirus (HPgV) and infect red colobus monkeys (Procolobus tephrosceles), red-tailed guenons (Cercopithecus ascanius) and an olive baboon (Papio anubis). We have designated these viruses SPgVkrc, SPgVkrtg and SPgVkbab, reflecting their host species' common names, which include reference to their location of origin in Kibale National Park, Uganda. SPgVkrc and SPgVkrtg were detected in 47% (28/60) of red colobus and 42% (5/12) red-tailed guenons, respectively, while SPgVkbab infection was observed in 1 of 23 olive baboons tested. Infections were not associated with any apparent disease, despite the generally high viral loads observed for each variant. These viruses were monophyletic and equally divergent from HPgV and pegiviruses previously identified in chimpanzees (SPgVcpz). Overall, the high degree of conservation of genetic features among the novel SPgVs, HPgV and SPgVcpz suggests conservation of function among these closely related viruses. Our study describes the first primate pegiviruses detected in Old World monkeys, expanding the known genetic diversity and host range of pegiviruses and providing insight into the natural history of this genus.


Bladder wall transplantation--long-term survival of cells: implications for bioengineering and clinical application.
Current bioengineered bladder wall substitutes include acellular scaffolds and grafts seeded with autologous cells. The transplanted cells on a seeded graft may regenerate and/or be replaced by cells of the patient's bladder. This may or may not be advantageous depending upon the underlying pathology. A theoretically perfect bioengineered graft would be intact bladder wall. To determine if such a graft is feasible and to study the cellular changes, we transplanted full-thickness bladder grafts from male inbred rats onto bladders of female syngeneic rats. Bladders were harvested at 1, 3, 6, 12, and 16 months after surgery and evaluated for histologic changes. Cell origin (male donor vs. female host) was determined with fluorescent in situ hybridization with unique probes for rat X and Y chromosomes. Urothelial hyperplasia, inflammation, and increased stromal thickness subsided down to control values by 6 months after surgery. At 16 months, graft muscle demonstrated persistence of male cells. On the other hand, graft urothelium was partially replaced by female host cells with a pattern suggestive of a hematogenous route rather than ingrowth from the host bladder. Bladder wall transplantation is feasible. The slow replacement of the transplanted urothelium and persistence of muscle may imply the same fate for engineered grafts.


[Bladder leiomyoma with extramural growth. An infrequent cause of pelvic pain].
A case of extravesical leiomyoma that had been incidentally discovered in a patient presenting with unspecific pelvic pain is described. A 51-year-old male patient consulted for unspecific pelvic pain. Patient evaluation disclosed a well-defined mass. A partial cystectomy was performed. Pathological analysis confirmed the diagnosis of leiomyoma of the bladder. Mesenchymal tumors of the urinary bladder are rare and account for 1%-5% of all bladder tumors. Leiomyoma is the most common benign non-epithelial tumor. The symptoms caused by this tumor type is varied, unspecific and in most of the cases depends on its location (intravesical, intramural or extramural). Treatment is by surgery, which achieves excellent results.


Serological evidence of enzootic bovine leukosis in the periurban dairy cattle production system of Al Ain, United Arab Emirates.
The serostatus of enzootic bovine leukosis (EBL) was determined at three dairy farms and the Al Ain Livestock Market (AALM), within the Al Ain region of Abu Dhabi, UAE. Of the 957 bovine sera tested by ELISA, 657 were from Holstein-Friesians from three dairy farms, and 300 from Bos indicus cattle at AALM. The chi-square homogeneity test (CSHT) and the Marascuilo multiple comparison procedure (MMCP) assessed the level of significance between the proportions of EBL-seropositive cattle (ESPC) across the study farms and AALM, and between the age groups at farms 1 and 3. Overall, the proportion of ESPC was 25.7% at dairy farms and AALM, 37.0% for farms and 1.0% for AALM. Furthermore, the proportions of ESPC at farms 1, 2 and 3 were 54.7%, 0.0% and 26.3% respectively, and statistically significant differences were seen across the farm/farm and farm/AALM comparisons, and between two age groups at farms 1 and 3. The 37-72-month-old age group showed the highest proportion of ESPC. This is the first serological evidence of EBL in the UAE. As previously reported, the ESPC are comparatively higher in dairy than Bos indicus cattle. Molecular and more extensive serological studies are needed to further corroborate the present data. Meanwhile, the UAE veterinary authorities will need to formulate national EBL control policies.


Trends in Helicobacter pylori resistance to clarithromycin: from phenotypic to genomic approaches.
For a long time Helicobacter pylori infections have been treated using the macrolide antibiotic, clarithromycin. Clarithromycin resistance is increasing worldwide and is the most common cause of H. pylori treatment failure. Here we review the mechanisms of antibiotic resistance to clarithromycin, detailing the individual and combinations of point mutations found in the 23S rRNA gene associated with resistance. Additionally, we consider the methods used to detect clarithromycin resistance, emphasizing the use of high-throughput next-generation sequencing methods, which were applied to 17 newly sequenced pairs of H. pylori strains isolated from the antrum and corpus of a recent colonized paediatric population. This set of isolates was composed of six pairs of resistant strains whose phenotype was associated with two point mutations found in the 23S rRNA gene: A2142C and A2143G. Other point mutations were found simultaneously in the same gene, but, according to our results, it is unlikely that they contribute to resistance. Further, among susceptible isolates, genomic variations compatible with mutations previously associated with clarithromycin resistance were detected. Exposure to clarithromycin may select low-frequency variants, resulting in a progressive increase in the resistance rate due to selection pressure.


[Lymphoepithelial organs of the pharynx in children after local immunomodulatory therapy].
The immunomorphology of adenoids and hypertrophied palatine tonsils in frequently ill children after treatment with herbal and microbial immunomodulators reflects the functional tension and interaction of both innate and adaptive immunity cells. The increase in the count of BCL-2-expressing cells is considered as possible dysregulation of the physiological processes of positive and negative selection via apoptosis.


Co-loading of coralyne and indocyanine green into adenine DNA-functionalized mesoporous silica nanoparticles for pH- and near-infrared-responsive chemothermal treatment of cancer cells.
Despite remarkable progress in the construction of functional mesoporous silica nanoparticles (MSNs) for cancer therapy, a multifunctional system with synergistic advantages over any single model remains encouraging. The objective of the present study was to develop a novel, simple and powerful nanotherapeutic system for the pH- and near-infrared (NIR)-responsive chemothermal treatment of cancer cells by co-loading coralyne and indocyanine green (ICG) into the adenine DNA (poly(A))-functionalized MSN. Coralyne, a type of planar alkaloid for cancer chemotherapy, can not only be loaded into the pores of MSN, but can also bind poly(A) and trigger the formation of non-Watson-Crick secondary structures, resulting in pore capping by the cooperative binding of poly(A) strands. Under low-pH or high-temperature conditions, non-Watson-Crick secondary structures were unstable, leading to the dehybridization of the cooperative binding structure and open-gate state. ICG, another cargo co-loaded into MSN, showed high efficiency for the conversion of NIR light into heat, which could provide the fundamental basis of hyperthermal therapy and promote pore opening upon NIR irradiation. In vitro studies using human hepatoma (HepG-2) cells demonstrated that this system could perform well in cellular acidic environment and under NIR irradiation, leading to a significant efficiency in controlled drug release and chemothermal cancer treatment. We believe that with these excellent features it should become a favorable nanoplatform for biomedical applications.


Effects of tumor-promoting phorbol diesters on neoplastic progression of Syrian hamster embryo cells.
The progression of normal Syrian hamster embryo cells to a neoplastic phenotype after treatment with a chemical carcinogen and continuous exposure to phorbol ester tumor promoters was studied in cell culture. Tumor promoters were able to rescue cell lines derived from individual carcinogen-treated colonies from a program of cellular senescence. In general, these cell lines did not grow in soft-agar medium or produce tumors in newborn hamsters at early passages but acquired these properties at later passages. These cell lines were used to study the temporal acquisition of a phenotypic characteristic of neoplastic rather than normal hamster embryo cells: the synergistic induction of the enzyme ornithine decarboxylase (ODC) by tumor promoter and serum growth factors (O'Brien, T. G., Saladik, D., and Diamond, L. Regulation of polyamine biosynthesis in normal and transformed hamster cells in culture. Biochim. Biophys. Acta, 632: 270, 1980). All cell lines that acquired neoplastic status with passage in culture exhibited the synergistic induction of ODC prior to their acquisition of the ability to either grow in soft-agar medium or produce tumors in newborn hamsters. Cell lines that responded to promoters with the synergistic induction of ODC accumulated greater levels of polyamines, especially putrescine, after promoter treatment than did normal cells. Tumor promoters did not affect the percentage of cells labeled with [3H]thymidine in preneoplastic cell lines, a finding similar to previous results in normal and neoplastic hamster cells. These studies demonstrate that tumor promoters can affect the early stages of neoplastic progression in carcinogen-treated Syrian hamster embryo cells and that a particular phenotypic property found in neoplastic hamster cells, the synergistic induction of ODC by tumor promoters and serum growth factors, is acquired by cell lines before they acquire neoplastic potential.


Barriers and Solutions to Advancing Emergency Medicine Simulation-based Research: A Call to Action.
Simulation technology has successfully improved patient safety and care quality through training and assessment of individuals, teams, and health care systems. Emergency medicine (EM) continues to be a leader and pioneer of simulation, including administration of simulation-based fellowships and training programs. However, EM simulation-based research has been limited by low rates of publication and poor methodologic rigor. The Society for Academic Emergency Medicine (SAEM) Simulation Academy is leading efforts to improve the quality of scholarship generated by the EM simulation community and to foster successful research careers for future generations of EM simulationists. Through a needs assessment survey of our membership and a year-long consensus-based approach, we identified two main clusters of barriers to simulation-based research: lack of protected time and dedicated resources and limited training and mentorship. As a result, we generated four position statements with implications for education, training, and research in EM simulation and as a call to action for the academic EM community. Recommendations include expansion of funding opportunities for simulation-based research, creation of multi-institutional simulation collaboratives, and development of mentorship and training pathways that promote rigor in design and methodology within EM simulation scholarship.


Molecular heterogeneity of pediatric choroid plexus carcinomas determines the distinctions in clinical course and prognosis.
Choroid plexus carcinomas (CPCs) are rare aggressive pediatric tumors of the brain with no treatment standards. Genetic profiling of CPCs is often confined to possible association with Li-Fraumeni syndrome, though only about a half of CPCs develop from syndromic predispositions. Whole-chromosome gains and losses typical of CPCs reflect genomic instability of these tumors, but only partially explain the aggressive clinical course. This retrospective study enrolled 25 pediatric patients with CPC, receiving treatment between January 2009 and June 2022. Molecular-genetic testing was performed for 20 cases with available tumor tissue and encompassed mutational status, chromosomal aberrations, and gene expression profiles. We analyzed several factors presumably influencing the outcomes, including molecular profiles and clinical parameters. The median follow-up constituted 5.2 years (absolute range 2.8-12.6 years). All studied CPCs had smooth mutational profiles with the only recurrent event being TP53 variants, either germline or somatic, encountered in 13 cases. Unbalanced whole-chromosome aberrations, notably multiple monosomies, were highly typical. In 7 tumors, chromosome losses were combined with complex genomic rearrangements: segmental gains and losses or signs of chromothripsis. This phenomenon was associated with extremely low 5-year survival: 20.0 ± 17.9% vs 85.7 ± 13.2%; P = .009. Transcriptomically, the cohort split into 2 polar clusters Ped_CPC1 and Ped_CPC2 differing by survival: 31.3 ± 17.8% vs 100%; P = .012. CPCs split into at least 2 molecular subtypes distinguished both genomically and transcriptomically. Clusterization of the tumors into Ped_CPC1 and Ped_CPC2 significantly correlates with survival. The distinction may prove relevant in clinical trials for dedicated and patient-oriented optimization of clinical protocols for these rare tumors.


[Rapid method for the determination of choline kinase activity. Choline kinase in marine invertebrates].
A quick sensitive test is suggested to determine choline kinase (EC 220.127.116.11). The method includes incubation of a substrate with enzyme in microvolume and separation of the formed phosphorylcholine from the initial choline by high-performance thin-layer silica gel chromatography. The procedure of determination is simple, reproducible and takes no more than 30 min. The method reveals high activity of choline kinase in some marine invertebrates. Certain features of the enzyme from the intestine of strongylocentrotus intermedius are described.


The Possibility of Therapeutic Drug Monitoring of the Most Important Interactions in Nursing Homes.
Therapeutic drug monitoring is a relevant tool in drug treatment of elderly patients. The aim of this study was to assess the possibility of therapeutic drug monitoring of the most important potential interactions in nursing homes. A material of prescribed drugs to 446 patients in three nursing homes in Bergen, Norway from a single day in March 2016 was analysed. Clinically relevant drug interactions (pharmacodynamic or pharmacokinetic) were identified and classified with Stockley`s Interaction Alerts. The most important interaction among several in each patient were ranked by recommended action > severity > evidence according to Stockley`s. The possibility of therapeutic drug monitoring of drug combinations involved in the most important interactions was retrieved from a database of all laboratories performing clinical pharmacology in Norway (the Pharmacology Portal). Two or more drugs were used by 443 (99.3%) of 446 patients. Three-hundred and eightyfour patients (86.1%) had > 1 interaction. About 95% of the most important interactions were pharmacodynamic. In 280 (72.9%) of these interactions, Stockley`s recommended adjust dose or monitoring. Among the 384 most important interactions, 93% involved one drug and 41% involved two drugs available for therapeutic drug monitoring. In this pilot study, therapeutic drug monitoring was possible in the majority of the most important interactions in Norwegian nursing homes. This option is of importance since adjust dose or monitoring were frequently recommended actions associated with these interactions.


Synaptic behavior and STDP of asymmetric nanoscale memristors in biohybrid systems.
We fabricate and characterize asymmetric memristors which show a very strong single-sided hysteresis. When biased in one direction there is hysteresis and in the opposite direction there is a lack of hysteresis. We demonstrate that this apparent lack is actually hysteresis on a much faster time-scale. We further demonstrate that this form of asymmetric behavior correlates very well to the asymmetric structure and function of an actual synapse. The asymmetric memristor device presented here is necessary to correctly implement spike-timing-dependent-plasticity STDP in mixed memristor/neuron hybrid systems as an artificial synapse. These devices show the required characteristics for implementing the asymmetric form of long-term potentiation (LTP) and long-term depression (LTD) of a synapse between two neurons, where symmetric memristor devices do not. Signals from a presynaptic neuron are sent via its axon across the synapse to the dendrite of a postsynaptic neuron. Postsynaptic neuron signals sent to subsequent neurons have an influence on the strength of any further presynaptic neuron signals received by the postsynaptic neuron across the synapse. These signals are grouped into spike triplets within the framework of STDP and, as we experimentally show here, can be implemented with asymmetric memristors, not standard symmetric memristors.


Dimensional analysis, spin freezing and magnetization in spin ice.
Dimensional analysis is shown to give an insight into the non-ergodic behaviour of spin ice below its apparent 'spin freezing' temperature. Expressions are derived for the temperature-dependent magnetic susceptibility that are found to be highly consistent with the previously reported field cooled and zero field cooled magnetization of the spin ice dysprosium titanate, Dy(2)Ti(2)O(7), as well as with the theory of a 'magnetolyte', including Debye-Hückel screening and Wien dissociation. The spin freezing is inferred to reflect the inability of the quasi-free magnetic charges or 'monopoles' that comprise the magnetolyte to fully screen an applied magnetic field on the timescale of an experiment. The apparent freezing temperature (T(f)≈0.65 K) is identified as the point where the Debye screening length becomes greater than the Bjerrum association distance for charge pairs. Combining these dimensional arguments with Onsager's theory of the Wien effect, it is shown that magnetization data at relatively high field (Snyder et al 2004 Phys. Rev. B 69 064414) may be used to estimate the elementary magnetic charge of spin ice, as well as the temperature-dependent monopole density. Evidence is presented of a non-equilibrium population of monopoles below T≈0.2 K. It is also shown how Onsager's microscopic theory of field-induced monopole pair separation naturally suggests the 'magnetization jumps' in Dy(2)Ti(2)O(7) observed at applied fields of the order of ∼ 0.1 T. It is concluded that the results of dimensional analysis, when combined with Onsager's theory, provide an accurate, albeit approximate, description of the properties of Dy(2)Ti(2)O(7), that could be improved by the development of a lattice theory of the Wien effect, or tested on other spin ice materials.


Ezrin regulates the expression of Mrp2/Abcc2 and Mdr1/Abcb1 along the rat small intestinal tract.
Multidrug resistance-associated protein 2 (MRP2)/ATP-binding cassette protein C2 (ABCC2) and multidrug resistance protein 1 (MDR1)/ABCB1 are well-known efflux transporters located on the brush border membrane of the small intestinal epithelia, where they limit the absorption of a broad range of substrates. The expression patterns of MRP2/ABCC2 and MDR1/ABCB1 along the small intestinal tract are tightly regulated. Several reports have demonstrated the participation of ERM (ezrin/radixin/moesin) proteins in the posttranslational modulation of MRP2/ABCC2 and MDR1/ABCB1, especially with regard to their membrane localization. The present study focused on the in vivo expression profiles of MRP2/ABCC2, MDR1/ABCB1, ezrin, and phosphorylated ezrin to further elucidate the relationship between the efflux transporters and the ERM proteins. The current results showed good correlation between the phosphorylation status of ezrin and Mrp2/Abcc2 expression along the gastrointestinal tract of rats and between the expression profiles of both ezrin and Mdr1/Abcb1 in the small intestine. We also demonstrated the involvement of conventional protein kinase C isoforms in the regulation of ezrin phosphorylation. Furthermore, experiments conducted with wild-type (WT) ezrin and a T567A (Ala substituted Thr) dephosphorylated mutant showed a decrease in membrane surface-localized and total expressed MRP2/ABCC2 in T567A-expressing vs. WT ezrin-expressing Caco-2 cells. In contrast, T567A- and WT-expressing cells both showed an increase in membrane surface-localized and total expressed MDR1/ABCB1. These findings suggest that the phosphorylation status and the expression profile of ezrin differentially direct MRP2/ABCC2 and MDR1/ABCB1 expression, respectively, along the small intestinal tract.


Mapping phosphoproteins in Neisseria meningitidis serogroup A.
To investigate the phosphorylation capability of serogroup A Neisseria meningitidis (MenA) and to implement our knowledge in meningococcal biology and in bacterial post-translational modifications, cell extracts were separated by 2-DE and 51 novel phosphoproteins were revealed by the use of the highly specific Ser/Thr/Tyr-phosphorylated proteins staining by Pro-Q Diamond and identified by MALDI-ToF/MS. Our results indicate that phosphorylation in MenA is comparable to that of other bacterial species. A first functional characterization of the identified modified proteins was also given, in order to understand their role in meningococcal physiopathology.


Limited Resuscitation With Fresh or Stored Whole Blood Corrects Cardiovascular and Metabolic Function in a Rat Model of Polytrauma and Hemorrhage.
We have recently shown that human whole blood stored at 4°C maintains hemostatic and platelet function. In this study, we compared restoration of hemodynamic, metabolic and hemostatic function after limited resuscitation with rat fresh whole blood, rat stored whole blood, or Lactated Ringers in traumatized rats. Rat whole blood was stored for 10 days at 4°C for evaluation of hemostatic function. Polytrauma was performed on isoflurane-anesthetized Sprague-Dawley rats (350-450 g) by damage to the intestines, liver, right leg skeletal muscle, and right femur fracture, followed by 40% hemorrhage. At 1 h, rats were resuscitated (20%) with either fresh whole blood (FWB), stored whole blood, 4°C for 7 days (SWB), Lactated Ringers (LR), or nothing. Blood samples were taken before and 2 h after trauma and hemorrhage to evaluate metabolic and hemostatic function. Whole blood stored for 10 days showed a significant prolongation in prothrombin time (PT) and activated partial thromboplastin time (aPTT), and fall in fibrinogen concentration, but no change in Maximum Clot Firmness or speed of clot formation. Platelet function was maintained until day 7 in storage, than fell significantly. Polytrauma and hemorrhage in rats led to a fall in arterial pressure, plasma bicarbonate, fibrinogen, and platelet function, and a rise in plasma lactate, PT, aPTT, and creatinine. Resuscitation with either FWB or 7 day SWB, but not LR, returned arterial pressure, plasma lactate and plasma bicarbonate to levels similar to control, but had no effect on the fall in fibrinogen or platelet function, or the rise in PT, aPTT, or creatinine. Hemostatic and platelet function of rat whole blood stored at 4°C is preserved for at least 7 days in vitro. Low volume resuscitation with SWB or FWB, but not LR, restores hemodynamic and metabolic function, but not the coagulopathy after severe trauma and hemorrhage.


A model-adjusted space-time scan statistic with an application to syndromic surveillance.
The space-time scan statistic is often used to identify incident disease clusters. We introduce a method to adjust for naturally occurring temporal trends or geographical patterns in illness. The space-time scan statistic was applied to reports of lower respiratory complaints in a large group practice. We compared its performance with unadjusted populations from: (1) the census, (2) group-practice membership counts, and on adjustments incorporating (3) day of week, month, and holidays; and (4) additionally, local history of illness. Using a nominal false detection rate of 5%, incident clusters during 1 year were identified on 26, 22, 4 and 2% of days for the four populations respectively. We show that it is important to account for naturally occurring temporal and geographic trends when using the space-time scan statistic for surveillance. The large number of days with clusters renders the census and membership approaches impractical for public health surveillance. The proposed adjustment allows practical surveillance.


The Effect of a Newly Designed Needle on the Pain and Bleeding of Patients During Oocyte Retrieval of a Single Follicle.
The purpose of this study was to evaluate the effect of needle type on pain and bleeding during oocyte pick-up (OPU). From May through November 2013, patients undergoing OPU from a single follicle without any analgesic treatment were including this study. Eligible patients (n=75) were randomized 1:1 to undergo the procedure with either a reduced needle (17 gauge body, 20 gauge tip; RN group) or a standard needle (19 gauge; SN group). Overall pain was assessed by patients using a visual analogue scale (VAS), and vaginal bleeding after the procedure was recorded. Fisher exact, t-test or Wilcoxon test were used, and p<0.05 was considered to be statistically significant. The percentage of mature oocytes was 86.5% in the RN group and 91.7% in the SN group. Pain during OPU was significantly lower in the RN group than in the SN group (mean VAS score±SD: 3.2±2.0 cm vs. 4.9±2.2 cm, p<0.01; mean±SD). The frequency of vaginal bleeding was also significantly lower in the SN group (26.3% vs. 48.6%; p<0.05). The frequency of bleeding in the RN group was also significantly lower than that in the SN group (26.3% vs. 48.6%; p<0.05). No significant differences were found between the two groups with regard to fertilization and pregnancy rates. The newly designed needle significantly reduced pain and vaginal bleeding associated with single-follicle OPU in patients receiving no analgesic treatment, in comparison with a standard needle. The RN had no adverse effect on the quality of retrieved oocytes.


Characterization of duck H5N1 influenza viruses with differing pathogenicity in mallard (Anas platyrhynchos) ducks.
A number of H5N1 influenza outbreaks have occurred in aquatic birds in Asia. As aquatic birds are the natural reservoir of influenza A viruses and do not usually show clinical disease upon infection, the repeated H5N1 outbreaks have highlighted the importance of continuous surveillance on H5N1 viruses in aquatic birds. In the present study we characterized the biological properties of four H5N1 avian influenza viruses, which had been isolated from ducks, in different animal models. In specific pathogen free (SPF) chickens, all four isolates were highly pathogenic. In SPF mice, the S and Y isolates were moderately pathogenic. However, in mallard ducks, two isolates had low pathogenicity, while the other two were highly pathogenic and caused lethal infection. A representative isolate with high pathogenicity in ducks caused systemic infection and replicated effectively in all 10 organs tested in challenged ducks, whereas a representative isolate with low pathogenicity in ducks was only detected in some organs in a few challenged ducks. Comparison of complete genomic sequences from the four isolates showed that the same amino acid residues that have been reported to be associated with virulence and host adaption/restriction of influenza viruses were present in the PB2, HA, NA, M and NS genes, while the amino acid residues at the HA cleavage site were diverse. From these results it appeared that the virulence of H5N1 avian influenza viruses was increased for ducks and that amino acid substitutions at the HA cleavage site might have contributed to the differing pathogenicity of these isolates in mallards. A procedure for the intravenous pathogenicity index test in a mallard model for assessing the virulence of H5/H7 subtype avian influenza viruses in waterfowl is described.


Endoanal ultrasound-guided surgery for anal fistula.
Outcomes following surgical treatment of patients with anal fistula are related to eradication of tracts and the internal opening. In this study, the results of surgery based on endoanal ultrasound (EAUS) findings were evaluated. A total of 102 patients with primary cryptogenetic anal fistula were prospectively examined with EAUS, using a 360-degree rotating 10-MHz probe, equipped with a three-dimensional (3-D) imaging system. Injection of hydrogen peroxide through the external opening was also used. Patients underwent operation on the basis of the EAUS findings. The agreement between findings from EAUS and from surgery was calculated. Clinical results were reported as treatment success, fistula recurrence, and fecal incontinence. Amongst 102 patients, the overall concordance between EAUS and surgical findings was 94.1 % for primary tracts, 91.2 % for internal openings, 96.1 % for secondary tracts, 100 % for abscesses, and 96.1 % for horseshoe tracts. Diagnostic accuracy was improved when hydrogen peroxide injection or 3-D imaging were used. Fistulotomy was performed in 46 patients (45.1 %), fistulectomy in 17 (16.7 %), fistulotomy plus seton placement in 19 (18.6 %), fistulectomy plus seton in 18 (17.6 %), and mucosal flap advancement in 2 (2.0 %). The operation was curative in 100 patients (98.0 %), and unsuccessful in 2 (2.0 %) due to recurrence of the fistula. Fecal continence was preserved in all patients. These data highlight the diagnostic accuracy of EAUS, particularly when hydrogen peroxide injection or 3-D imaging are used. Basing our surgical decision making on EAUS findings allowed us to carry out curative operations in a significantly large number of patients; the recurrence rate was very low. The accurate EAUS assessment of the relationship between fistulas and sphincters has been the main factor in choosing a sphincter-saving surgical procedure, avoiding fecal incontinence.


Disease awareness in patients with COPD: measurement and extent.
Patient awareness of COPD refers to knowledge and acceptance of the disease and its treatment. Although it is relevant to management and outcomes, the disease awareness of patients is poorly investigated, and no validated questionnaires are currently available. We aimed to develop the novel Disease Awareness in COPD Questionnaire (DACQ), which was validated in relation to demographic and clinical features, in patients participating in the SATisfaction and Adherence to COPD Treatment (SAT) study. DACQ was developed according to a list of items regarding the patient's knowledge, acceptance, and perception of COPD as well as of treatment needs. The questionnaire was validated by assessing internal structure and consistency, correlations with other patient-reported outcomes, and stability over time. Furthermore, the extent of disease awareness of patients enrolled in the SAT study was assessed by using DACQ, and correlations with demographic and clinical features were evaluated. DACQ was composed of four domains. Overall reliability and stability over time were adequate; correlations between DACQ and other tools measuring different constructs (ie, treatment satisfaction, illness perception, impact of COPD symptoms on daily life, and dyspnea severity) were, as expected, more limited. In the enrolled patient sample, a suboptimal level of disease awareness (<70%) was detected, especially in terms of disease acceptance and perception. Disease knowledge was positively associated with COPD severity, while the impact of symptoms on daily life was negatively associated with disease acceptance, awareness of treatment needs, and overall awareness. DACQ proved to be a reliable tool to assess awareness in COPD patients. Awareness of COPD patients need to be improved. ClinicalTrials.gov ID# NCT02689492.


Helium Cold Atmospheric Plasma Causes Morphological and Biochemical Alterations in Candida albicans Cells.
(1) Background: Previous studies reported the promising inhibitory effect of cold atmospheric plasma (CAP) on Candida albicans. However, the exact mechanisms of CAP's action on the fungal cell are still poorly understood. This study aims to elucidate the CAP effect on C. albicans cell wall, by evaluating the alterations on its structure and biochemical composition; (2) Methods: C. albicans cells treated with Helium-CAP were analyzed by atomic force microscopy (AFM) and Fourier transform infrared spectroscopy (FTIR) in order to detect morphological, topographic and biochemical changes in the fungal cell wall. Cells treated with caspofungin were also analyzed for comparative purposes; (3) Results: Expressive morphological and topographic changes, such as increased roughness and shape modification, were observed in the cells after CAP exposure. The alterations detected were similar to those observed after the treatment with caspofungin. The main biochemical changes occurred in polysaccharides content, and an overall decrease in glucans and an increase in chitin synthesis were detected; (4) Conclusions: Helium-CAP caused morphological and topographic alterations in C. albicans cells and affected the cell wall polysaccharide content.


School Ijime (Bullying) Experience Is a Possible Risk Factor for Current Psychological Distress among Science City Workers: A Cross-Sectional Study in Tsukuba, Japan.
Early-life adversities are considered to have long-term impact on health. There have been many studies regarding the experience of being bullied and its harmful psychological influence, but such influence on workers remains to be investigated in Japan. We therefore aimed to examine the prevalence of workers with experiences of being bullied or bullying others during childhood and adolescence and to clarify the relation between those experiences and current psychological distress. A cross-sectional study using an anonymous self-report web questionnaire was conducted in November 2017. The study population was 19,481 workers belonging to the Tsukuba Science City Network, and we analyzed the data of 6,015 participants (3,715 men and 2,300 women, aged between 20 years and 59 years). The percentages of participants with experiences of being bullied were 51.5% for men and 56.2% for women; those with experiences of bullying others were 36.5% of men and 29.4% of women. Relations between experiences of bullying and psychological distress were assessed using Chi-squared tests. Logistic regression analyses with psychological distress as an objective variable and experiences of bulling as explanatory variables were performed using those who had no bulling experiences as control. In both men and women, experiences of being bullied were significantly associated with psychological distress after adjustment socioeconomic factors (the odds ratios were 1.26 for men (95% CI = 1.05-1.52) and 1.72 for women (95% CI = 1.39-2.13)). Consideration of past social experiences, such as being bullied, is useful for mental health management among workers reporting psychological distress.


Last resort beta-lactam antibiotics for treatment of New-Delhi Metallo-Beta-Lactamase producing Enterobacterales and other Difficult-to-Treat Resistance in Gram-negative bacteria: A real-life study.
Novel last resort beta-lactam antibiotics are now available for management of infections due to New-Delhi Metallo-Beta-Lactamase (NDM) producing Enterobacterales and non-fermenters with Difficult-to-Treat Resistance. However, data regarding the use of imipenem-cilastatin-relebactam (IMI-REL), cefiderocol (CFD) and ceftazidime-avibactam plus aztreonam (CAZ-AVI-ATM) are scarce in real-life settings. This study aimed to describe the use of last resort beta-lactam antibiotics, the microbiology and the outcome, in patients hospitalized in a tertiary hospital. We conducted a monocentric observational cohort study from 2020/01/01, to 2022/08/31. We screened all patients admitted to Nimes University Hospital who have received ≥ 1 dose of last resort beta-lactam antibiotics during the study period, using the Pharmacy database. We included patients treated with IMI-REL, CFD and CAZ-AVI-ATM. The primary endpoint was the infection-free survival rate. We also calculated rates of microbiological and clinical cure, recurrent infection, death and adverse events. Twenty-seven patients were included in the study and 30 treatment courses were analyzed: CFD (N=24; 80%), CAZ-AVI-ATM (N=3; 10%) and IMI-REL (N=3; 10%). Antibiotics were used in 21 males (70%) and 9 females (30%) with a median age at 65-year-old [50-73.5] and a median Charlson index at 1 [0-2]. Almost all the patients had ≥ 1 risk factor for carbapenem resistant bacteria, a half of them was hospitalized for severe COVID-19, and most of antibiotic courses (N=26; 87%) were associated with ICU admission. In the study population, the probability of infection-free survival at day-90 after last resort beta-lactam therapy initiation was 48.4% CI95% [33.2-70.5]. Clinical failure rate was at 30%, microbiological failure rate at 33% and mortality rate at 23%. Adverse events were documented in 5 antibiotic courses (17%). In details, P. aeruginosa were mainly treated with CFD and IMI-REL, S. maltophilia with CFD and CAZ-AVI-ATM, A. baumannii with CFD, and NDM producing-K. pneumoniae with CAZ-AVI-ATM and CFD. After a treatment course with CFD, CAZ-AVI-ATM and IMI-REL, the probability of infection-free survival was 48% CI95% [10.4-73.5], 33.3% CI95% [6.7-100], 66.7% CI95% [30-100], respectively. Use of last resort beta-lactam antimicrobials in real-life settings was a safe and efficient therapeutic option for severe infections related to Gram-negative bacteria with Difficult-to-Treat Resistance.


Comparative transcriptome and metabolome analyses revealed quality difference between beauty tea processed through indoor withering and outdoor solar withering.
Withering is the first processing procedure of beauty tea, and there are few reports on the impact of withering methods on the quality of beauty tea and its regulatory mechanisms. Through comparison of fresh tea leaves (FT) with the leaves after indoor natural withering for 18 h (IWT-18) and outdoor solar withering for 6 h (OWT-6), which were collected at the end of the two withering processes, 17 282 and 13 984 differentially expressed genes (DEGs) were respectively screened and 267 and 154 differential metabolites (DMs) were respectively identified. The coexpression network revealed that a large number of DEGs and DMs were enriched in phenylpropanoid, flavonoid, and adenosine triphosphate binding cassette (ABC) transporter pathways, and the number of DMs and DEGs in IWT-18 versus FT exceeded that in OWT-6 versus FT. Both withering methods promoted a significant increase in content of phenylalanine and upregulation of β-glucoside expression in the phenylpropanoid metabolism pathway. Five theaflavin-type proanthocyanidins in the flavonoid synthesis pathway were more significantly accumulated in FT versus IWT-18 than in FT versus OWT-6. Meanwhile, both withering methods can affect the ABC transporter pathway to promote the accumulation of amino acids and their derivatives, but different withering methods affect different ABC transporter families. Outdoor withering with more severe abiotic stress has a greater impact on the ABCG family, whereas indoor withering has a more significant effect on the ABCC family. Sensory evaluation results showed that the dry tea of IWT-18 was slightly better than that of OWT-6 because of the longer withering time and more thorough substance transformation. In conclusion, the formation of honey flavor in beauty tea may be closely related to the DEGs and DMs in these three pathways. Our research provides theoretical data support for further revealing the mechanism of quality formation during the withering process of beauty tea. © 2023 Society of Chemical Industry.


A cell-free system for the study of alpha-amylase synthesis in barley aleurone layers.
1. A cell-free system capable of alpha-amylase synthesis has been obtained from the aleurone layers of germinating barley. 2. This system requires potassium chloride, sucrose and an amino acid mixture in order to function. The crude preparation does not require calcium chloride. Chloramphenicol inhibits alpha-amylase synthesis as indicated both by increase in measurable enzyme activity and incorporation of l-[U-(14)C]glutamic acid.


Molecular hybridization between rat liver deoxyribonucleic acid and complementary ribonucleic acid.
RNA (cRNA) was synthesized in vitro on a template of rat liver DNA and its hybridization with rat liver DNA was studied by using the nitrocellulose-filter method. Sonication of the DNA diminished its apparent capacity to hybridize with RNA by about 50%. This is not due to cross-linkage of DNA molecules, because it could be shown that less than 2% of the sonicated DNA was cross-linked. The effect is due instead to the small size of the sonicated DNA molecules. Below a single-stranded molecular weight of 5x10(5) the DNA showed a progressive loss of capacity to hybridize with decrease in molecular weight. Evidence is presented suggesting that the apparently diminished capacity of the DNA to hybridize is due to loss of hybridized DNA from the membrane filters. When cRNA at concentrations of up to 25mug/ml is annealed with sonicated total DNA, an apparent hybridization saturation value is found at which about 2.5% of the DNA is hybridized with RNA. Increasing the cRNA concentration tenfold brought about the hybridization of a second component of the DNA approximately equal in amount to the first. The renaturation of rat liver DNA was studied by measuring the fall in the extinction at 260nm and two different components of renaturation were observed within the reiterated fraction of DNA. By hybridizing cRNA with different fractions of rat DNA the two components of the hybridization curve are shown to correspond to the two components of the renaturation curve. The conclusion is drawn that at a cRNA concentration of 250mug/ml most of the reiterated fraction of rat liver DNA is hybridized after annealing for 16h under standard conditions (0.30m-sodium chloride-30mm-sodium citrate at 65 degrees C). Even with such a high cRNA concentration little or no hybridization of the slowly renaturing DNA fraction occurs. It is suggested that the most highly reiterated DNA component is poorly transcribed in vitro.


Evidence against extracellular exposure of a highly immunogenic region in the C-terminal domain of the simian immunodeficiency virus gp41 transmembrane protein.
The generally accepted model for human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein topology includes a single membrane-spanning domain. An alternate model has been proposed which features multiple membrane-spanning domains. Consistent with the alternate model, a high percentage of HIV-1-infected individuals produce unusually robust antibody responses to a region of envelope, the so-called "Kennedy epitope," that in the conventional model should be in the cytoplasm. Here we show analogous, robust antibody responses in simian immunodeficiency virus SIVmac239-infected rhesus macaques to a region of SIVmac239 envelope located in the C-terminal domain, which in the conventional model should be inside the cell. Sera from SIV-infected rhesus macaques consistently reacted with overlapping oligopeptides corresponding to a region located within the cytoplasmic domain of gp41 by the generally accepted model, at intensities comparable to those observed for immunodominant areas of the surface component gp120. Rabbit serum raised against this highly immunogenic region (HIR) reacted with SIV envelope in cell surface-staining experiments, as did monoclonal anti-HIR antibodies isolated from an SIVmac239-infected rhesus macaque. However, control experiments demonstrated that this surface staining could be explained in whole or in part by the release of envelope protein from expressing cells into the supernatant and the subsequent attachment to the surfaces of cells in the culture. Serum and monoclonal antibodies directed against the HIR failed to neutralize even the highly neutralization-sensitive strain SIVmac316. Furthermore, a potential N-linked glycosylation site located close to the HIR and postulated to be outside the cell in the alternate model was not glycosylated. An artificially introduced glycosylation site within the HIR was also not utilized for glycosylation. Together, these data support the conventional model of SIV envelope as a type Ia transmembrane protein with a single membrane-spanning domain and without any extracellular loops.